Recommendations for the Successful Captive Breeding of North American River Otters (Lontra canadensis)

Transcription

1 Recommendations for the Successful Captive Breeding of North American River Otters (Lontra canadensis) An Internship Report Presented to the Faculty of the College of Science and Mathematics Colorado State University - Pueblo Pueblo, Colorado In Partial Fulfillment Of the Requirements for the Degree of Master of Science in Biology By Allison Kema Rose Haar Colorado State University Pueblo December, 2015

2 CERTIFICATE OF ACCEPTANCE This Internship Report Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in Biology By Allison Kema Rose Haar Has Been Accepted By the Graduate Faculty of the College of Science and Mathematics Colorado State University Pueblo APPROVAL OF INTERNSHIP REPORT COMMITTEE Graduate Advisor (Dr. Brian Vandenheuvel) Date Committee Member (Dr. Lee Anne Martínez) Date Committee Member (Dr. Annette Gabaldón) Date Graduate Director (Dr. Dan Caprioglio) Date

3 Dedication To my family, especially my amazing parents, Pat and Deb: you have been so supportive while I undertook this project. I couldn t ask for more caring or supportive parents. You have given me every opportunity to follow my dreams and to find new ones. Thank you. To my wonderful nieces, Kasey, Meghan, and Caitlyn: you have loved me unconditionally, even on the days when I said, I can t play. I have to go work on my paper. There was no complaining, just understanding. I hope you know that you can do anything you want in life. And always remember, life can change in an instant, it may not go how you planned, but family will always love and support you, no matter what happens. I am so grateful to be your aunt. Thank you for reminding me how to have fun when life gets too stressful. v

4 Acknowledgments I would like to thank all of the professors in the CSU-Pueblo biology department for helping me through the years, especially my committee members Dr. Brian Vandenheuvel, Dr. Lee Anne Martinez, and Dr. Annette Gabaldón. Also, thank you to Dr. Jack Seilheimer. I would like to offer a special thanks to Dr. Jeff Smith and Dr. Paul Chacon for their hours of help on formatting the flow of the paper and creating useable results from the mass of information with which we started. I would like to thank the Regional Access to Graduate Studies (RAGE) office for their dedication to proper grammar and punctuation throughout the writing process. They know more about river otters now than they ever wanted to know, but they were invaluable. Thank you Roy Jo Sartin and Erin DeCuir. Thank you to Marilyn McBirney, Melanie Pococke, and everyone at the Pueblo Zoo. It was wonderful working with you all through this process. vi

5 Abstract The North American river otter (NARO) (Lontra canadensis) is a semi-aquatic mammal found in the wild throughout North America, and captive at many institutions throughout the world. The Association of Zoos and Aquariums (AZA) captive NARO population represents an important feature species that attracts many patrons to zoological institutions. NAROs are rarely taken from the wild, so captive breeding programs are being utilized to increase the captive population. These captive breeding programs are overseen by the AZA to ensure the genetic diversity of the captive populations, and to encourage proper care and management of the animals. Captive breeding, however, is not always successful. In fact, captive breeding of mustelidae (members of the weasel family) has been a learning experience for all involved; as seen by the easy success of the Black-footed ferret (Musetla nigripes), the hit or miss success of the North American river otter (Lontra canadensis), and the lack of success with the sea otter (Enhydra lutris). This research project was initiated to identify factors, such as geographic origin of the breeding pair and zoological practices, that contribute to successful captive breeding. Observations of the Pueblo Zoo (Pueblo, CO) NAROs and their zoological practices were conducted as part of this study. This information was used to create a survey, which asked specific information about the captive NAROs, and the zoological practices that impact their captivity. The survey was then sent to all AZA institutions that house and attempt breeding with their captive NAROs. Results from the 30 responding AZA institutions indicate that having two NAROs from similar geographic locations, both in proximity and latitude of origin, aids in successful captive breeding. No single vii

6 zoological practice was demonstrated to have a significant impact on successful captive breeding. Despite low response rate, the evidence suggests that successful captive breeding is more likely to occur between NAROs from similar locations, and that, as expected, providing a healthy environment that meets all biological needs may aid in successful captive breeding. viii

7 Table of Contents Page Dedication Acknowledgments Abstract List of Figures List of Tables v vi vii xi xiii Introduction 1 Background and Significance History of the North American River Otter (Lontra canadensis) 8 Physical and Reproduction Characteristics 24 Resources and Breeding Recommendations 39 Summary of Significance and Justification of the Research Question 53 Hypotheses and Specific Aims 56 Study Design Internship and Observations 58 Data Collection 66 Analysis 68 Results Geographic Origins 70 Zoological Practices 74 Discussion and Future Studies 85 Conclusions and Recommendations 97 ix

8 Literature Cited 99 Appendices A. Ethological Study of the Pueblo Zoo North American River Otters (Lontra 108 canadensis) during the 2010 Breeding Season B. Institutions Contacted for the Research Project 120 C. Surveys 136 D. Data 154 E. Pedigree Analysis 194 F. Internship Journals 216 x

9 List of Figures Page Figure 1. Historic Range of the NARO, Before the Mid-1800s 9 Figure 2. Physical Map of North America 10 Figure 3. Biomagnification of PCBs Through the Food Chain 12 Figure 4. NARO Range circa Figure 5. Distribution of the NARO circa Figure 6. Geographic Boundaries of the NARO Subspecies 19 Figure 7. Distribution of the NARO circa Figure 8. Comparison of the Number of NAROs Entering the Captive AZA 26 Population Each Year Figure 9. Age Class Structure of the NARO Captive Population in AZA 27 Accredited Institutions Figure 10. Management Hierarchy of AZA Groups and Committees 43 Responsible for the Creation of Animal Welfare Documents Figure 11. Schematic Drawing of the Pueblo Zoo NARO on and off-exhibit 61 Areas Figure 12. Diagram of the NARO Indoor Holding Area 63 Figure 13. Map Comparing the Distances of Origination of Male and Female 71 NAROs of Successful Breeding Pairs Figure 14. Map Comparing the Distances of Origination of Male and Female 72 NAROs of Unsuccessful Breeding Pairs xi

10 Figure 15. Mean Comparison of the Distance of Origination between Male 73 and Female NAROs of Successful and Unsuccessful Breeding Pairs Figure 16. Mean Comparison of the Difference in Latitude of Origination of 74 the Male and Female NAROs of Successful and Unsuccessful Breeding Pairs Figure 17. Comparison of the Type of Daily Supplementation Given to 77 NAROs at Successful and Unsuccessful Institutions Figure 18. Comparing the Location of Dens Available to the Female NAROs 78 from Successful and Unsuccessful Institutions Figure 19. Comparison of the Type of Pen Available to Female NAROs at 79 Successful and Unsuccessful Institutions Figure 20. Comparing the Seasonal Availability of Nesting Materials 81 Available to Female NAROs of Successful and Unsuccessful Institutions Figure 21. Comparison of the Percent of Breeding Pairs for Successful and 82 Unsuccessful Institutions Figure 22. Comparison of the Amount of Daily Interaction between the 83 Zookeepers and the NAROs of Successful and Unsuccessful Institutions Figure 23. Responses to the Final Opinion Based Survey Question Whether 82 or Not the Zookeepers Feel They Have an Impact on Captive Breeding xii

11 List of Tables Page Table 1. Foods Given as Part of the Daily Diet for NAROs by Successful and 75 Unsuccessful Institutions Table 2. Enrichment Foods Given to NAROs by Successful and Unsuccessful 76 Institutions Table 3. Comparison of the Type of Nesting Material Made Available to Female 80 NAROs at Successful and Unsuccessful Institutions xiii

12 Introduction Captive breeding The purpose of captive breeding is to maintain a population of animals, whether captive or wild (Miller et al. 1996). The goal of any captive breeding program is to create a group of animals that can be reintroduced to the wild as quickly as possible (Miller et al. 1996). This means minimizing the amount of time each generation is held in captivity in order to reduce the occurrence of adaptation to captivity and to reduce the chances of selecting for characteristics that would cause a disadvantage in the wild (Allendorf 1993; Lacy 1994; Miller et al. 1996; Lynch & O Hely 2001). There are many pros and cons to captive breeding programs. Successful programs, like that of the Black Footed Ferret (Mustela nigripes), bring awareness to the animals and their causes (Miller et al. 1996). This awareness can, in turn, bring about changes to policy to improve the environment and help wild populations. Black-footed ferrets and North American river otters (Lontra canadensis) owe some of their successful reintroductions and relocations to policy changes that created protection for the animals and protected space for them (i.e. being listed by the Endangered Species Act of 1973) (U.S. Fish and Wildlife Service 2013). Captive breeding can also be a very effective tool when alternatives, such as habitat recovery or relocation, are unavailable in the short term (Snyder et al. 1996). Unfortunately, captive breeding programs can inadvertently cause genetic changes that may reduce sustainability in the wild (Allendorf 1993; Lacy 1994; Miller et al. 1996; Lynch & O Hely 2001; Robert 2009). Even under the best circumstances and with the best intentions, characteristics in behavior, morphology, and physiology that are 1

13 beneficial to captive environments can be unintentionally selected over the preferred wild traits (Allendorf 1993; Miller et al. 1996; Lynch & O Hely 2001; McPhee 2003; Robert 2009). A few examples: captive breeding programs normally do not have any predators through which the animal would learn avoidance; captive breeders can also select for high juvenile growth rates, which may not be in line with normal reproductive rates (Lynch & O Hely 2001). Using these individuals can cause an irreversible loss of genetic diversity (Fraser 2008). Small populations, whether wild or captive, also have a more difficult time adapting to genetic changes and deleterious loads. Any small change can quickly cause larger problems (Lacy 1994; Lynch & O Hely 2001). Fortunately, a wild population can readapt to the natural environment, but it may take many generations of reduced fitness in the meantime (Lynch & O Hely 2001). This means that there is little leeway for the amount of change that occur within a captive population. After this threshold has been passed, the reintroduced animals will fail to be successful in the wild environment (Lynch & O Hely 2001). Captive breeding has been well studied and we now know that captive populations, especially those destined for breeding and reintroductions, should ideally start with at least 20 founders, and have 7-12 progeny produced per founder, if possible (Lacy 1994). To ensure diversity and reduce the chance of total loss due to unforeseen catastrophe, the population should be spread around several institutions (Lacy 1994; Miller et al. 1996; Bryant et al. 1999). Reintroduction of captive bred animals should only begin after the captive population has reached its planned capacity, and this should be evaluated after each breeding season (Lacy 1994). It is also important to note that the chance of having negative characteristics and adaptations in a captive breeding 2

14 population can be reduced by increasing the population size (Allendorf 1993; Miller et al. 1996; Bryant et al. 1999). Captive breeding is a tool to maintain threatened and endangered species, though it is a second choice over improvements to the natural habitat and affected ecosystem (Miller et al. 1996). Additionally, captive breeding is expensive and requires a lot of space (Miller et al. 1996). A captive breeding program should always be well thought out before any animals are taken into custody. Although it can be expensive and difficult to maintain, captive breeding has its benefits and should not be overlooked as a very important way to maintain endangered and captive species. The musetelidae (weasel family) has many examples of captive breeding programs, including: black-footed ferrets (Mustela nigripes), North American river otters (Lontra canadensis), and sea otters (Enhydra lutris). Black-Footed Ferret The Black-footed ferret (Mustela nigripes) is an excellent example of successful captive breeding, not only of a mustelid, but for any program. Like the North American river otter, the Black-footed Ferret (BFF) was a dominant part of its ecosystem, the prairie, until hunting almost extirpated the entire population (Miller et al. 1996). In fact, the BFF was thought to be extinct until a small population of animals was found in South Dakota in 1981 (Murray 1987; Clark 1994, 1997; Miller et al.1996). Unfortunately, this population was further decimated by an outbreak of sylvatic plague; all known individuals were lost. In 1986, 18 new individuals were found in Wyoming. Of these few BFFs, only seven animals were successfully bred to establish today s entire captive breeding and wild populations (Clark 1994, 1997; U.S. Fish and Wildlife Service 2013). 3

15 This represents a major bottleneck and an extreme reduction in the genetic fitness for any population (U.S. Fish and Wildlife Service 2013). There were many political issues when the BFF breeding program began (Murray 1987; Miller et al. 1996). For example, there has always been a lot of controversy over who should be in charge of the population and the captive breeding program. Federal agencies, state agencies, the Association of Zoos and Aquariums (AZA), non-profit organizations, and ranchers are all involved in the decision-making process for the BFF (Murray 1987; Clark 1996, 1997; Miller et al. 1996). Having to work through the chainof-command has created many extra problems for the breeding program. There is often a power struggle that can make implementing changes difficult (Miller et al. 1996). Thankfully, everyone has managed to move forward through the years, and the program has continued to be successful. BFFs are currently bred at the Conservation and Research Center in Wyoming, and at five AZA institutions: Cheyenne Mountain Zoological Park, Louisville Zoological Gardens, Smithsonian Conservation Biology Institute, Phoenix Zoo, and Toronto Zoo (Clark 1994, 1997; Miller et al. 1996; Association of Zoos and Aquariums 2013a). These institutions work together to create the most genetically diverse population possible and to continue to breed healthy animals that can be reintroduced into their natural habitat. This goal includes dealing with concerns about their small population size and their ability to breed well in captivity. Artificial insemination has been used with success in this breeding population (U.S. Fish and Wildlife Service 2013). There has been a lot of concern over the years about the small population size and the likelihood that the BFFs in captivity would successfully breed. A lot of care has been 4

16 taken to reduce the amount of inbreeding, and its subsequent reduction of fitness, for the captive BFF population (Jiménez et al. 1994). Inbreeding has been shown to reduce the success of reintroduced animals, even if they seem fine in a captive environment (Murray 1987; Jiménez et al. 1994). The original BFF captive population was so small that inbreeding was inevitable. Best practices and a solid breeding plan have led to a large and thriving population that does not seem to show any long-term issues due to inbreeding. Photoperiod, specifically lengthening daylight, is known to play a key role in BFF reproduction. Wild populations breed in the spring, while captive populations can be bred into the summer months (Miller et al. 1996). Knowing the natural rhythms of the wild populations has allowed the captive breeding program to create a more supportive environment for successful captive breeding. This also creates a situation where the captive animals have a reduced exposure to situations and rhythms that are drastically different from what they will experience once they are living in the wild. There is current interest in having captive-bred individuals exposed to wild situations before release, if possible, to better acclimate them to what they may face (Fraser 2008). For example, BFFs that are preconditioned in outdoor pens (i.e., exposure to dogs) have a much higher survival rate than those who are cage-reared and then released into their new wild environment (Clark 1997). The ultimate goal of the BFF captive breeding program would be an effective population of 1,500 BFFs to maintain at least 80% genetic diversity. This can only happen if wild populations are established (Miller et al. 1996). The captive breeding program has had success over the years, with more than 8,000 kits born, and is on its way 5

17 to this goal (U.S. Fish and Wildlife Service 2013). New animals are released into the wild each year. Sea Otter Sea otters (Enhydra lutris) provide a unique study among mustelids, and a completely different captive breeding story from the BFF. They are truly marine mammals, not necessarily coming ashore at any point in their lives (Kruuk 2006; Shirihai 2006). They are unique among otter species in several respects: their ability to drink sea water, dive to great depths, their extremely dense fur, and their use of tools to access food (Kruuk 2006). Sea Otters are the largest member of mustelidae, they have true flippers for back feet, while the front feet have partly retractable claws. Sea otters are very mild mannered and incredibly vulnerable to human exploitation (Kruuk 2006). Like the NARO and the beaver, their pelts are highly prized. Sea otters were extirpated from much of their home range by 1820, due to hunting (Kruuk 2006); but they have been legally protected since This led to an initial increase in population size, but they have never regained their pre-1800s numbers. Part of this is due to their slow reproductive cycle and susceptibility to pollution in the ocean (Kruuk 2006). Sea otters are polygynous, where one male breeds with several females (Kruuk 2006; Shirihai 2006). There is a loosely defined breeding season. Females give birth from December to February in California and from May to June in Alaska; although pups can be born year-round (Larson et al. 2003; Kruuk 2006; Shirihai 2006). Gestation lasts around six and a half months. Females undergo a delayed implantation for two to three months followed by approximately four months of actual gestation (Larson et al. 2003; Kruuk 2006). 6

18 Sea otters are not sexually mature until two years of age and some sea otters do not start breeding until five years of age (Kruuk 2006). All of these factors can complicate captive breeding efforts. It has been suggested that pups born to younger mothers have a decreased chance of survival due to the mother s inexperience, among other factors (Kruuk 2006). It is unknown if sea otters are induced ovulators (i.e., ovulation only occurs after the baculum (penile bone) stimulates the ovaries to release an ova) like many other mustelids (Larson et al. 2003) Captive breeding of sea otters has not been very successful. Only two institutions (The Seattle Aquarium and Oregon Zoo) have successfully bred and raised sea otters past two years of age. Pups have been born at other institutions, but they have not survived to sexual maturity (two years of age) (Larson et al. 2003). There are known subspecies of sea otters inhabiting their natural range, as well as genetic stock variations within the subspecies (Kruuk 2006). This has the potential to further complicate mate choices and reduce the chance of a successful outcome when attempting captive breeding. Although captive breeding could be a great benefit to both captive and wild populations of sea otters, thus far they have proved very difficult to breed. Understanding the reproductive patterns of sea otters and how to create a more successful captive breeding program will greatly increase the chance of success. Summary These two mustelid family examples showcase both the possibilities of success and the difficulties that come with any breeding program. The North American river otter is no different. These animals are currently bred to increase both captive and wild populations. There have been successes at some institutions, while other institutions seem 7

19 to have an ideal environment with no success. Understanding not only how a captive breeding program can and should be run, but also the individual species, can increase the chance that future captive breeding programs will find success. History of the North American River Otter (Lontra canadensis) Historic Habitat and Range The North American river otter (Lontra canadensis) is a semi-aquatic mammal that until the mid-1800s, ranged throughout most major waterways and coastal areas across North America (Fig. 1). The North American river otter (NARO) is found in both freshwater and marine habitats and is an important riparian keystone species for several reasons. First, these otters are good indicators of a healthy ecosystem, because they are at the top of the food chain. They have no natural predators in the water and very few on land (Macdonald & Mason n.d.). Second, they often prey on rough fish, which are larger fish species that are not commonly eaten by humans, and are not sought after as sport fish (Hill 1994). Rough fish eaten by NAROs include several types of carp, gar, and drum. Lastly, they are very sensitive to water pollution and are easily affected when any part of the ecosystem is polluted. Because they are a keystone species, when NARO numbers decline, the effects to the surrounding ecosystem are larger than would be expected for a single species due to their critical role (van Wieren 2006). 8

20 Figure 1. Historic Range of the North American River Otter (Lontra canadensis). Before the mid- 1800s, their range encompassed much of North America north of Mexico (Reed-Smith 2012a). Extirpation Due to Human Activity Before the mid-1800s, NAROs were found throughout North America. As human habitation expanded, it caused a significant impact on every environment human beings encountered either by physically changing the landscape or via overhunting of various animals leading to low population numbers. Although the NARO is a mobile species, there are certain geographic boundaries they are unlikely to cross. The major waterways and mountain ranges throughout North America are shown in Figure 2. NAROs are excellent swimmers, so the waterways would provide an opportunity for distribution. Mountain ranges, on the other hand, could be a hindrance to distribution, as the NAROs are unlikely to cross an entire range. 9

21 Figure 2. Physical map of North America showing the major mountain ranges and waterways. North American river otters (Lontra canadensis) are very mobile and could easily use waterways for dispersal. (Oxford Atlas of the World 2012) As their name suggests, NAROs are dependent on water for survival, but not all waterways and mountains provide adequate food and habitat. Due to changes in their historic habitats, there was a decline in the NARO population from the mid-1800s to the mid-1900s; the NARO was extirpated from Colorado in the early 1900s (Murray 1987). Several sources list the following factors that caused a decline in NARO populations: destruction of habitat, trapping and hunting, and pollution (Raesly 2001; Nilsson and Vaughn, as cited in Reed-Smith 2012a). Destruction of natural habitat Ever-growing human populations are continually encroaching on natural NARO habitats, which can cause otters to migrate elsewhere. Human beings have made many 10

22 changes to the landscape as they have moved further inland, such as clearing land, draining marshes, and channelizing streams (Nilsson and Vaughn, as cited in Reed-Smith 2012a; Mason and Macdonald n.d.) to create space for homes, businesses, roads, and agriculture. In addition, waterways are often diverted for agricultural purposes, which changes the natural flow and composition of the source waters (e.g. chemical composition, turbidity, and flow rate). These changes have in turn, reduced the amount of riparian habitat available for NAROs. Trapping and hunting Trapping and hunting are major factors in NARO extirpation. Beginning in the late 1800s, their fur was highly prized as the standard for durability and quality, by which all other furs were judged (Melquist et al. 2003). As with other furs at the time, otter pelts were used to make coats and fur trim. Even today, otter pelts are highly valued for their durability and high-quality (Hill 1994; Melquist et al. 2003). As recently as February 2013, the average price for an otter pelt at auction was $104, with the highest priced pelt selling for $170. Beaver pelts average only $33 a pelt (Fur Harvesters Auction 2013). Hunting of otters, whether directly or when caught as bycatch in beaver traps, is thought to be the major cause of the NARO extirpation, nearly destroying most populations (Melquist and Dronkert 1987). Pollution Another human factor negatively affecting NARO populations well into the 20 th century is pollution, which affects the water, the food, and the otters themselves (Kruuk 2006). NAROs have a high sensitivity to pollution and are easily harmed by it (Hill 1994). Not only can the otters be directly affected by pollutants in the water, but the 11

23 aquatic food they ingest will lead to an increase in their intake of pollutants (Hill 1994). Biomagnification is the natural process of concentrating nontoxic levels of pollutants into toxic levels as the compound moves up the trophic levels of a food chain (Langenbach 2013). The result is a much higher concentration of toxic compounds for top predators. Many pollutants are also lipophilic compounds, meaning they will not dissolve in water, and are, therefore, more likely to be concentrated through natural biomagnifications. One example that affects NAROs is water contaminated by polychlorinated biphenyls (PCBs), organic pollutants that are not easily removed from the environment. PCBs are known toxic compounds and have already caused a decline in the otter population in Oregon (Hancox 1992; Foster-Turley n.d. a). The amount of PCBs ingested by otters due to its biomagnification in the food chain is diagramed in Figure 3. Figure 3. Example of the biomagnification of PCB through the food web and the final concentration that affects North American river otters (Lontra canadensis) (Reproduced from New York Department of Environmental Conservation 2013). There is a known negative correlation between PCB concentration and physical condition of the animal (Kruuk 2006). It is thought that PCBs cause sterility in developing pups (Hancox 1992), which would further harm the population. Another example of pollution 12

24 is acid mine waste, such as that created in the early 1900s. This waste directly caused poor water quality from runoff and dumping, which has added to population losses (Macdonald and Mason n.d.; Colorado Department of Wildlife 2003). Effects on natural environment due to their extirpation As a top predator, NAROs are responsible for keeping many fish populations, especially rough fish, under control in their natural habitat (Hill 1994). In one study, Melquist and Hornocker (1983) found that when an abundant source of food diminished, or other prey became more available, otters either moved to a new location or changed their feeding habits and selected the most available prey. NAROs will typically change feeding sites or will change prey before they consume all of one species. Without the NARO, many larger fish species could potentially overtake the smaller species and throw off the community structure. The presence of NAROs also plays a key role with beaver populations. NAROs are closely associated with (Kruuk 2006) and have a facultative commensalism relationship with beavers (Foster-Turley, as cited in Reed-Smith 2012a). This relationship allows NAROs to use beaver dens as their own, whether or not the beaver is still present. If the beaver abandons its den, the dam will fall into disrepair. As it falls apart, the surrounding area is changed, just as the area in which the new beaver dam is built changes. These changes to the environment create an ever shifting community of various species depending on the beaver dam s effects on the stream. Also, due to their close relationship, when beavers are hunted, NAROs are often caught in the traps, so both populations suffer losses. 13

25 NARO populations contribute to understanding the health of the ecosystem as well, because of their role as keystone species. When there is a steady, healthy population of NAROs, it indicates that there is an abundant food source and plenty of suitable habitats. If the otter population declines, it is an indicator that something has changed in that area, and the problems should be addressed. Dramatic changes in their environment should also be expected once they are gone (van Wieren 2006). Their place as a keystone species requires that thought be given to the potential impact and changes that might occur by reintroducing them into their natural historic habitats. Current Habitat and Range Conservation efforts As outlined above, changes to the natural landscape and increased pollution resulting from an increasing human presence led to the extirpation of NAROs from most of their natural habitat (Melquist and Dronkert 1987; Reed-Smith 2012a). As a result of these devastating population declines, conservation efforts were initiated, beginning with trapping and hunting regulations, improvements to water quality, as well as reintroductions. When these conservation efforts began, the range of NAROs, circa 1977, was only 25% of their historic range throughout North America, as shown in Figure 4 (Reed-Smith 2012a). This included occupying approximately 33% of their historic range in the United States (Reed-Smith 2012a). 14

26 Figure 4. North American river otter (Lontra canadensis) range circa 1977, occupying approximately 25% of their historic range (Melquist et al. 2003). Improvements in hunting and trapping regulations Part of the conservation efforts to protect NAROs included legal protection to reduce hunting and trapping (Mason and Macdonald n.d.). This realization led to hunting and trapping seasons that are highly regulated, if they are even allowed. Each year, individual states evaluate their populations to determine if there are enough otters to warrant a trapping or hunting season. Each state sets their own limits for the length of seasonal activities, as well as the ways in which the otters may be trapped or killed, and a limit to the number of otters one hunter can kill. According to Hill (1994), 17 states completely ban hunting and trapping the river otter, 27 states allow trapping, and four states and two Canadian provinces allow hunting. New United States federal regulations, which follow the Convention on International Trade in Endangered Species (CITES) of 15

27 Wild Flora and Fauna guidelines, ensure otter pelts are not leaving the country from which they are captured, without being tagged and reported (Boyle 2006; Polechla n.d.). The continuing regulation of these activities will support the growth of NARO populations and the lasting improvements to the ecosystems. Early conservation efforts for beavers, which were also decimated by hunting, inadvertently created improvements for otter populations. Restocking and reintroduction of beavers, which occurred from the 1920s to 1950s, had a positive impact on otter populations (Foster-Turley as cited in Reed-Smith 2012a; Hill 1994). Increased beaver populations means the creation of more beaver dams, which increase wetland areas, providing more fishing areas for otters. NAROs also like to use beaver lodges as den sites, which are used more often than any other type of den (Melquist and Hornocker 1983). Beaver traps historically have killed many otters, but these must now be no-kill traps to ensure otters can be released. Improvements to water quality Major improvements to water quality are another source of conservation for NAROs (Raesly 2001). The Clean Water Act and other wetland conservation efforts have noticeably changed water quality in the United States. Originally, these changes were made to protect human health, but they have had many benefits for native wildlife as well. Pollution has decreased, which has a positive impact on the ecosystem. Improved water quality has created an environment for a healthier food supply for the NARO as well. 16

28 Initial population improvements Changes to trapping and hunting regulations, as well as environmental policy changes, resulted in an increase in NARO populations and expansion of their range, so that by 1988 they occupied approximately 50% of their total historic range (Reed-Smith 2012a) (Fig. 5). Improvements in water quality have also aided in the NARO recovery (IUCN Otter Specialist Group 2011). Conservation efforts will help the continued expansion of wild NARO populations and help promote healthy ecosystems throughout North America. Figure 5. Distribution of the North American river otter (Lontra canadensis) in Dots represent reintroduction sites; stars represent verified sightings (Melquist et al. 2003). 17

29 Current Status in the Wild Possible Subspecies There has been extensive debate about the number of, and the geographic boundaries that define NARO subspecies. For the purpose of this project, seven subspecies were considered when comparing captive otter origins. The following list of subspecies is found in the Association of Zoos and Aquariums NORTH AMERICAN (Nearctic) RIVER OTTER (Lontra canadensis) Husbandry Notebook Section 1, Chapters 1-6 (Reed-Smith 2012a), which defines the boundaries of each subspecies. The numbers correspond to Figure 6, a map showing the basic geographic boundaries of each subspecies: 1. L. c. canadensis: Newfoundland, Illinois, Wisconsin, Michigan, Minnesota, eastern seaboard states, Ontario, Quebec, New Brunswick, and Nova Scotia 2. L. c. lataxina: southeastern US, central plains, and Gulf of Mexico states 3. L c. pacifica: Alaska, Yukon, and Northwest Territories; British Columbia, Alberta, Saskatchewan, Manitoba, and parts of Ontario; northern California, Oregon, Washington, Idaho, Montana, Wyoming, North Dakota, South Dakota, most of Minnesota, parts of Colorado, northern Utah, and northern Nevada 4. L. c. sonora: Arizona, Nevada, southeastern California, southern Utah, and New Mexico 5. L. c. mira: Alexander Archipelago, Alaska 18

30 6. L. c. periclyzomae: Queen Charlotte Islands, Windfall Harbor, Admiralty Island, and Savok Bay area in Alaska 7. L. c. kodiacensis: Kodiak and Afognak Islands in Alaska Figure 6. Geographic boundaries of the North American river otter (Lontra canadensis) subspecies as defined in the NORTH AMERICAN (Nearctic) RIVER OTTER (Lontra canadensis) Husbandry Notebook Section 1, Chapters = L. c. canadensis; 2= L. c. lataxina; 3= L c. pacifica; 4= L. c. sonora; 5= L. c. mira; 6= L. c. periclyzomae; 7= L. c. kodiacensis (Reed-Smith 2012a). The possibility of NARO subspecies has been debated often, but never formally studied. Without genetic analysis, the subspecies definitions use trivial differences in physical appearance and some geographic barriers. For these subspecies, differences in physical appearance consist of noticeable difference in size; cranial shape and details; tails that are proportionally longer or shorter; length of the hind foot; and whether there is hair on the inferior surface of feet and webbing (Reed-Smith 2012a). Obvious geographic boundaries would include islands that are reasonably separated from the mainland, mountain ranges, and ranges interrupted by major cities. 19

31 Arguments in support of otter reintroductions The current status of NAROs in the wild varies depending on the state or territory in question. NAROs have large, thriving populations along the east coast and in Canada, but they are still recovering in the interior states and the arid southwest (Melquist et al. 2003). Reintroductions, defined as returning a species to areas where their populations have been extirpated, are the best method to return an endemic species, such as the NARO, back to its natural habitat (Miller et al. 1999; Melquist et al. 2003; van Wieren 2006; Ralls n.d.). Reintroductions have occurred in 21 states across the US since 1976 (Raesly 2001) and have been fairly successful (Kruuk 2006). The reasons for reintroductions vary by state, but include the potential for harvest, aesthetic value, cultural significance, ecological importance, and commercial value (Erickson and McCullough 1987). The reintroduction programs have been successful because of previous efforts to improve natural habitats and ecosystems as a whole. Attempting reintroductions would be a waste of resources if there was nothing to support the reintroduced animals. High habitat quality is essential to increase the chance that a healthy population will survive (Griffith et al. 1989; Ralls n.d.). For NAROs this was achieved in earlier decades with trapping and hunting regulations, as well as improvements to water quality (Raesly 2001). In the past, carnivorous species have proved more difficult to reintroduce to their natural habitat, but once established, they can have many positive effects on the ecosystem (Miller et al. 1999). This is seen with NAROs because of their keystone role in riparian habitats and their ability to affect fish community structure (Foster-Turley n.d. a; Melquist et al. 2003). 20

32 Reintroduction programs are also dependent on captive breeding programs (Wisely et al. 2003). A good captive breeding program will reduce the chance of issues related to inbreeding (Jiménez et al. 1994). A good reintroduction program must find a way to reduce the loss of genetic variability, caused by inbreeding, which can cause overall decreased fitness of the animal, including disease susceptibility, lowered fecundity, high infant mortality, and reduced growth rates (Caro & Laurenson 1994; Jiménez et al. 1994; Wisely et al. 2003). Programs must also choose individuals who are likely to survive in the wild; these are often the otters who have not adapted to captive living (Wisely et al. 2003). Reintroduction in Colorado In Colorado, NAROs were designated a state endangered species in 1975 (Colorado Division of Wildlife 2003). Based on the subspecies as defined by Reed-Smith (2012a), Colorado has two potential subspecies within its borders: L. c. lataxina and L. c. sonora. Between 1976 and 1991, more than 114 NAROs were reintroduced at five sites in Colorado (Colorado Department of Wildlife 2003). This included otters from all over North America, such as Minnesota, Newfoundland, and Alaska. The goal of Colorado reintroduction efforts is to maintain a population at the species level, not necessarily the subspecies level (Colorado Division of Wildlife 2003), which is why choosing native subspecies was not a priority. Since NAROs are so versatile and mobile, having a subspecies that is not native to Colorado should not be a problem, as long as there are populations containing otters of similar subspecies. In fact, it is believed that due to the mobility of NAROs, use of subspecies alone when choosing otters for reintroductions is inappropriate (Serfass et al. 1998). 21

33 Because they are a highly mobile, elusive species, it is difficult to know the exact numbers of wild otters. NARO signs -- tracks, scat, slides, dens, prey remains, and scent mounds -- are used in addition to direct sightings when evaluating if a healthy population has been established (Colorado Division of Wildlife 2003). Based on sightings and signs since the reintroductions, established breeding populations have been found along the Colorado River, the Piedra-San Juan River complex, the Dolores-San Miguel River drainages, the Gunnison River, lower Yampa River, and the Green River (Colorado Division of Wildlife 2003). Because self-sustaining populations have been established, the reintroductions are considered a success in Colorado (Griffith et al. 1989), and as of the fall of 2012 the NARO status has been downgraded to threatened (Colorado Parks and Wildlife 2013). Ultimately, reintroductions have attempted to recreate the historic NARO distribution. The success of reintroductions in the United States and Canada can be seen in Figure 7, a map from demonstrating a bridge of NARO ranges connecting the northern distributions and southern distributions. Dots represent reintroduction sites while stars represent verified sightings. 22

34 Figure 7. Distribution of the North American river otter (Lontra canadensis) in , occupying approximately 75% of their historic range. Dots represent reintroduction sites; stars represent verified sightings (Melquist et al. 2003). Improvements of ecosystems due to the return of otters Another benefit to NARO reintroductions is their role as a keystone riparian species, and their use as indicators of a healthy environment (Foster-Turley n.d. a). A healthy NARO population keeps fish numbers in balance. Once NARO populations improve, the surrounding ecosystems also begin to change for the better. Otters could be considered beneficial to natural populations of fish, as well as game fisheries, because they prey on slower-moving nongame and forage fish (Polechla n.d.). Removing these fish from the environment removes the game fishes competition for food. Another benefit to wetlands and riparian habitats is the conservation status that they will have when NAROs are allowed back into their natural habitats and both they and the habitats protected (Melquist et al. 2003). 23

35 Physical and Reproductive Characteristics General Species Description and Characteristics The North American river otter (NARO) (Lontra canadensis) is a semi-aquatic mammal inhabiting both freshwater and marine habitats. The NARO head is broad and dorsoventrally flattened (Peterson 1966). Their small eyes and ears are placed towards the top of the head, which allows full use of the senses while most of their body remains underwater (Baitchman & Kollias 2000). Because they are mouth-oriented feeders, the prominent whiskers allow NAROs to capture prey more easily in murky water (Mason n.d.). The NARO body is long and cylindrical, giving it a streamlined nature (Baitchman & Kollias 2000). The tail is long and tapered, which increases the streamlined profile in the water and decreases the agility of NAROs on land. Because it comprises one-third of the animal s total length, the tail also provides stability and power for short bursts of rapid locomotion when swimming (Hill 1994). NAROs have short, powerful legs, and their feet are fully webbed for swimming. NARO toes have tufts of hair on the underside to increase dexterity, while the claws at the end of the toes aid in catching prey (IUCN Otter Specialist Group 2011). Male NAROs average 122 cm (48 inches) in length, but may reach 137 cm (54 inches) (Larivière and Walton 1998; Kruuk 2006). Males weigh an average of 11.3 kg (25 pounds), though some have been measured at 15 kg (33 pounds) (Hill 1994; Kruuk 2006). Females are slightly smaller than males, approximately 112 cm (44 inches) long and weighing an average of 8.6 kg (19 pounds) (Hall 1981; Hill 1994; Kruuk 2006). The 24

36 sexes are easily distinguished: males have a baculum (penile bone) and females have four mammae on the upper chest (Hill 1994). NARO fur is short, very dense, and glossy (IUCN Otter Specialist Group 2011). The color ranges from very dark on the dorsal side to light beige or gray on the chin and throat (Baitchman & Kollias 2000). The fur is comprised of two different types, one of which provides waterproofing and the other provides the primary means of insulation. Subcutaneous fat located at the base of the tail and on the rear legs provides the secondary source of insulation (Baitchman & Kollias 2000). NAROs can live an average of ten years in the wild, but may reach years of age in captivity. NAROs normally breed from ages 3-14 for males and 2-10 for females, although both sexes have been known to breed up to age 17 in captivity (Association of Zoos and Aquariums 2013b; Reed-Smith 2012a). The number of NAROs entering the captive population of the Association of Zoos and Aquariums (AZA) has steadily increased in the past, mostly due to the addition of rehabilitation and nuisance animals from the wild (Fig. 8) (Association of Zoos and Aquariums 2013b). 25

37 Number of Animals Total Wild born Captive born Unknown Year Figure 8. Comparison of the number of North American river otters (Lontra canadensis) housed in the Association of Zoos and Aquariums captive population, New individuals are added to the population from wild caught individuals or through captive breeding programs (Association of Zoos and Aquariums 2013b). To maintain wild populations, successful captive breeding should be encouraged and increased to ensure there will be new individuals available for reintroductions. Also important to the captive population is the ratio of males to females, especially those of breeding age. The age structure below shows the number of male and female NAROs in each age class of the current AZA captive population (Fig. 9). 26

38 Figure 9. Age structure of the potentially breeding population of North American river otters (Lontra canadensis) in the Association of Zoos and Aquariums captive population; data through September Most of the population fall within the breeding age. The red line indicates the projected population in 2017 (Association of Zoos and Aquariums 2013b). The age structure shows that sex ratio skews slightly male, but there is a stable base of individuals who are reproductively mature. The red line indicates the expected population in It shows that the population has some breeding individuals, but the numbers are notably reduced. The number of individuals in these lower breeding age groups needs to be increased, which can be achieved through captive breeding and the import of wild animals (Association of Zoos and Aquariums 2013b). The otters in AZA institutions are bred and held for captive purposes only. There are separate programs for otters destined for reintroductions. The breeding strategies and problems are similar for both types of captive populations. 27

39 Zoological Practices Daily diet NAROs are primarily piscivorous, although they could be considered an aquatic generalist; NAROs will consume almost anything they can find (Kruuk 2006; Reed- Smith 2012a). Larivière and Walton (1998) note that NAROs choose fish that are abundant and close to shore in inverse relation to the prey s swimming ability. Prey are captured with a quick lunge or ambush. The NARO rarely exerts the energy required for a long chase (Larivière and Walton 1998). Also important to the NARO diet are crustaceans (crayfish), amphibians (frogs), invertebrates, birds, reptiles, and some small mammals (Kruuk 2006; Reed-Smith 2012a). Some vegetation may be eaten during summer, but plants do not make up a considerable amount of the regular diet (Peterson 1966). Nutrition guidelines for Association of Zoos and Aquariums institutions Daily diet: According to the nutritional guidelines set forth by the International Union for the Conservation of Nature (IUCN)/ Species Survival Commission (SSC) Otter Specialist Group, and adopted by the AZA, NAROs have a high metabolic rate, digesting their food in only one to four hours. Therefore, otters in captivity should be fed at least twice a day, although three to four times a day is preferred (Henry et al. 2012). During feedings, scattering the food throughout the exhibit and offering live prey encourages natural hunting and foraging behaviors (Owens et al. 2009). NARO nutritional needs have not been thoroughly studied, so the house cat (Feliz catus) is used as a basis for nutritional guidelines (Henry et al. 2012). In fact, a commercial feline chow is often used as a staple NARO food product (Owens 2012). No matter what formal nutrition program 28

40 is designed, every NARO should receive a daily diet that accounts for each otters natural preferences, physical conditions, weight, amount of exercise, environment, the season, and actual nutrient requirements (Association of Zoos and Aquariums Small Carnivore TAG 2009). Fish should make up the bulk of the regular diet, preferably offered daily. The fish should be fresh and low in thiaminase and fat (e.g., capelin, smelt, herring, and trout). Nutritionally complete dry and wet cat foods, as well as horse meat or beef-based products, round out the main portion of the daily diet (Reed-Smith 2012b). Fresh fruits and vegetables should also be included in the regular diet. The vegetation increases bulk and adds necessary roughage. Hard items, such as rib bones, crunchy foods, and whole prey, aid in NARO dental care and offer a change in the daily diet (Owens et al. 2009; Henry et al. 2012). The daily diet should include a variety of items to stimulate the otters and to decrease the likelihood there will be a nutritional deficiency (Foster-Turley, n.d. b). Enrichment: The overall goal of enrichment is to offer a varying environment for NAROs to reduce the possibility of abnormal repetitive behaviors (ARBs). Mason (1991) defines an ARB as a behavior pattern that is repetitive, invariant, and has no obvious goal or function. ARBs are potentially harmful to captive animals and should be reduced or avoided if possible. For this reason, items should also be added to enrich interest and variety in the daily NARO diet. There are many different types of potential enrichment categories, some of which include: food type, delivery of food, environmental changes/additions, toys or manipulatable items, scents, and sounds (Association of Zoos and Aquariums Small Carnivore TAG 2009; Reed-Smith 2012c). Examples of 29

41 enrichment items include ice blocks, carcasses, brooms, balls, and new substrates. Randomizing feeding times, scattering and hiding the daily diet, and offering live foods are also forms of enrichment (Owens et al. 2009). Training is an important form of enrichment that serves several purposes for captive NAROs. Training is useful for husbandry and medical care, and creates a relationship of trust between the keepers and the NAROs (McKay 2009; Reed-Smith 2012c). Otters are highly intelligent, curious animals, so training is usually accomplished fairly easily. At AZA institutions, the keepers use positive reinforcement, such as a small piece of food, when the animal completes the required task (McKay 2009; Reed-Smith 2012c). The food reward that is given for performing the correct action can be used to give the otter a portion of their daily diet or as a supplement to the daily diet. Training for husbandry purposes involves teaching the animal to separate from enclosure mates and to enter/leave the exhibit on command. Training for medical purposes teaches NAROs to stand on a scale; enter a transport enclosure; allow for shots without being sedated; and allow visual inspection of the coat, paws, tail, and teeth (Owens et al. 2009). Training also helps prevent ARBs from occurring or from becoming worse if already present. As previously stated, ARBs are repetitive, invariant behaviors, with no obvious goal or function (Mason 1991). These behaviors are often related to suboptimal living conditions, which can be as minor as a lack of daily stimulation, or, like a coping mechanism, can be induced by frustration (Mason et al. 2007). It can be noted in the ethological study, found in Appendix A, that Thelma, the older female NARO at the Pueblo Zoo, would often swim in circles whenever someone stood in front of her on- 30

42 exhibit enclosure. This is the only potential ARB exhibited by the captive NAROs at the Pueblo Zoo. The keepers try to create a stimulating environment to avoid these potentially harmful behaviors. Supplements: The AZA suggests some nutritional supplementation should be considered for captive NAROs. At a minimum, captive NAROs should receive vitamin B1 and vitamin E daily (Reed-Smith 2012a). These are often given in the form of thiamin E paste, a mix of the two vitamins in an easy to dispense paste. This supplementation is recommended because nutrient loss, especially with vitamin E, can occur when fish is stored, frozen, and thawed (Reed-Smith 2012b). The AZA also recommends a multivitamin supplement to ensure all nutritional needs are being met. Families and groups NAROs are very social animals. In the wild, the basic social grouping is called a family and consists of a female and her juvenile offspring, called pups (Melquist and Hornocker 1983; Kruuk 2006). A few older females, who may or not be related, may join this female grouping and act as helpers for the pups (Melquist and Hornocker 1983). Other common groupings include groups of all males, groups of all females, solitary males, solitary females, and single male-female pairs during breeding season (Owens et al. 2009). The amount of resources available influences the type of grouping (Owens et al. 2009). Captive environments offer more flexible opportunities for groupings because the resources are always abundant. The Association of Zoos and Aquariums Small Carnivore Taxonomic Advisory Group (TAG) (2009) recommends the following groupings in captivity: multiple males, a male-female pair, one male and multiple females, or multiple 31

43 pairs. The Pueblo Zoo had two males and one female, which is not a grouping on the recommended list, but it has worked well for them. In 2013 they added a second female, which creates multiple pairs of NAROS, a recommended grouping. This situation has been accepted by all four otters. Behavior and communication NAROs are active at night and during crepuscular hours year-round, although they become more diurnal during the winter months and in undisturbed areas (Hill 1994; Boyle 2006). NAROs do not migrate, but they will move in response to food availability (Boyle 2006). Varying the times of day captive otters are fed, and providing enrichment can offer enough new activities for the captive NAROs to be more active during the day, which is when patrons are allowed to view the animals. NAROs are incredibly agile in the water, moving with an undulating motion or paddling with their limbs (Hill 1994; Larivière and Walton 1998). They are less agile on land, but still able to move quickly when necessary by running or bounding. They make use of slopes and icy areas by sliding along on their bellies (Peterson 1966). Play behavior is seen in pups and juvenile NAROs in the wild, and is prevalent in captive animals, in part because all of their basic needs have been met by the institution, meaning that they can expend less energy on foraging (Melquist and Dronkert 1987; Melquist et al. 2003). Play behavior includes sliding, wrestling, chasing the tail, playing with captured prey, and juggling pebbles or sticks while swimming (Larivière and Walton 1998). Many of these behaviors were observed in the captive NAROs at the Pueblo Zoo (Appendix A). 32

44 To communicate, NAROs use olfactory and auditory signals. Olfactory signals include scent markings, made via defecation, urination, anal sac secretions, and musk from scent glands. In the wild, these signals are used by otters to indicate the effective range of each individual. NAROs are not territorial, but scent marking does help individuals minimize contact to avoid any potential issues (Melquist et al. 2003). During the breeding season, scent markings increase and are even more important because they allow the males to more easily find a female who is receptive (Polechla n.d). These qualities are important in captivity as well; because they allow the female NARO to make known that she is receptive for breeding. Auditory communication comes in many different forms: shrill chirp, soft chuckle, scream, caterwaul, snarl, growl, hissing bark, shrill whistle, low purring grunt, and alarm call (explosive snort) (Liers 1951; Larivière and Walton 1998; Melquist et al. 2003; IUCN Otter Specialist Group 2011). Most of these vocalizations are used for regular communication, at play, and when travelling. Snarls, barks, and screams can be heard when animals are fighting and the caterwaul is only made by the female during copulation. Thelma, the older female NARO at the Pueblo Zoo, made some of the more alarming calls, such as screams, when she was unreceptive to the male advances during the breeding season (Appendix A). Range NAROs can travel great distances over any terrain (Melquist and Hornocker 1983). Although they can travel easily, they tend to stay within a homerange that is approximately 155 km² (60 mi²) (Hill 1994). Within a NARO s homerange are sites known as activity centers. These sites have adequate food and shelter and act as central 33

45 areas of activity for NAROs (Mason n.d.). These activity centers are important because NAROs travel following their food sources. There is very little territoriality usually centered on securing food sources, and occasionally include breeding areas dominated by a male (Murray 1987). Liers (1951) notes that travel is more common during the breeding season; most likely acting as a way to distribute the population and ensure breeding. Den and nest sites Because they are so mobile, NAROs will use many different den sites throughout the year. As they follow the availability of food to new locations, they will find new den sites. Dens are often chosen for convenience, although those that provide protection and seclusion are preferred (Melquist and Hornocker 1983). NAROs do not excavate their own dens, and will use any available structure, such as tree logs, natural formations, and dens made by other animals. They will enlarge a den site as needed (Liers 1951; Melquist and Hornocker 1983). Most often, NAROs in the wild use beaver (Castor canadensis) dams, whether actively inhabited or abandoned (Melquist and Hornocker 1983). Riparian habitats that provide many different foods and vegetative cover are the preferred habitat where NAROs will search for a den. When they are preparing a natal nest, females will choose dens near small headwater branches or streams that lead to major waterways (Hill 1994). The more secluded location provides more protection for the new mother and her pups. In captivity, several dens are usually provided for the NAROs, with guidelines suggesting at least one den per otter (Owens et al. 2009; Foster-Turley n.d. b). The Pueblo Zoo provides a holding area that is only available to the females, which contains a den (~ 30 x20 x15 ) that only they can enter. The size, shape, and type of den are all 34

46 determined by the institution based on guidelines and their particular NAROs. Nesting materials (the materials used to line the den to provide a nest for the new pups) are always provided by institutions with captive populations (Association of Zoos and Aquariums Small Carnivore TAG 2009; Owens et al. 2009; Appendix D). Sexual Characteristics Male NAROs Male NAROs normally reach sexual maturity around age two, although generally they are not reproductively successful until five to seven years old (Liers 1951). The baculum (penile bone) continues to grow in length until age three, and to gain weight until age six, at which point it is the correct size to induce ovulation in females (Liers 1951; Baitchman & Kollias 2000; Reed-Smith 2012a). Reproduction in the wild stops at approximately age ten. Captive NAROs differ from wild otters for the age range of successful reproduction. According to Reed-Smith (2012a) the youngest captive male NARO to successfully breed was less than one year and the oldest was 17. This is possible because all basic needs are being met in a captive environment, so NAROs can expend more energy on reproduction than they would in the wild. Female NAROs Females can mature as early as months of age, although they are usually not successful breeders until around two years of age (Association of Zoos and Aquariums Small Carnivore TAG 2009; Reed-Smith 2012a). The youngest female NARO to successfully breed in an AZA institution was just under two years of age, and the oldest NARO to reproduce successfully in captivity was 11 years of age. Though they are physiologically reproductive at an earlier age, female NAROs average five years of 35

47 age at their first successful reproduction (Reed-Smith 2012a). Female NAROs are induced ovulators, meaning they need an appropriately sized baculum to release the ova for fertilization (Association of Zoos and Aquariums Small Carnivore TAG 2009; Reed- Smith 2012a). Breeding season Female NAROs are monestrous, meaning they go through estrous for breeding only once per year. The breeding season lasts from November to May. Research has shown that the breeding season varies with latitude, with NAROs at low latitudes breeding earlier than those at high latitudes (IUCN Otter Specialist Group 2011; Reed- Smith 2012a). This may have an effect on successful captive breeding because many times the NAROs will be housed outside of their natural latitude, and paired with NAROs from dissimilar latitudes. It is also assumed that photoperiod plays a part in this process, most likely affecting the onset of estrous in females (Reed-Smith 2012a). Estrous lasts days, unless mating occurs, with peaks of maximum receptivity occurring every three to six days (Liers 1951; Reed-Smith 2012a). Many institutions rotate having their males and females together during this time to mimic wild conditions. During peak receptivity females will seek out a male and make copulation as easy as possible. When only mildly receptive, females may completely reject any male advances (Liers 1951). These behavior changes can be seen in the Pueblo Zoo NAROs (see Appendix A). NAROs are polygamous animals, meaning females and males will mate with several different individuals in one breeding season. In the wild this is possible because of their high mobility. In captivity this is achieved by housing multiple pairs together and 36

48 by changing the individuals that are housed together by transferring them to different institutions (Association of Zoos and Aquariums 2013b). Copulation characteristics Successful NARO copulation can occur on land, but is more likely to occur in the water. Successful copulation lasts a minimum of 15 minutes, but may last up to 70 minutes (Larivière and Walton 1998; Reed-Smith 2012a). The male approaches the female from behind and grabs her neck with his teeth. The copulation act is vigorous and females will often caterwaul during the act. The NAROs will copulate for an extended period followed by a period of rest or foraging. Copulation will continue in this manner several times a day and on consecutive days (Melquist and Dronkert 1987; Reed-Smith 2012a). If the female is not receptive to the male, she will not tolerate his presence and will scream and run aggressively towards him. All of these behaviors have been seen in the Pueblo Zoo NAROs (Appendix A). In captivity, there is an opportunity to make the most out of the female s peak receptivity periods by rotating the males on and off-exhibit with the female (Owens et al. 2009). This is especially useful when there are multiple males, and has the added benefit of reducing possible fights between the sexually aggressive males. This situation is called seasonal separation and has been implemented by the Pueblo Zoo (Appendix A). Gestation and birth NAROs undergo a process called delayed implantation, whereby the fertilized egg stops developing at the blastocyst stage (Hamilton and Eadie 1964; Kruuk 2006). It then floats freely in the uterus for about eight months before implanting in the uterus to finish development (Reed-Smith 2012a). Once the blastocyst implants, gestation takes 37

49 approximately days, giving a total gestation of around 12 months (Larivière and Walton 1998; Kruuk 2006; Reed-Smith 2012a). There is no specific known reason for delayed implantation in this species; however, it may allow for breeding over a longer period of time, the synchronization of parturition (the action of giving birth) and breeding, or an increase in potential mates for lactating females who are restricted to their natal dens (Reed-Smith 2012a). When it is time to give birth, the female NARO will find a secluded area for her natal den. For captive NAROs, the housing institution needs to provide a space for the female to make her natal den, which should be separated from the males. Parturition normally occurs in late winter to early spring, depending on latitude. Reed-Smith (2012a) recognizes that the timing of breeding and parturition positively correlates with the captive female s birth location, not her current geographic location. This means that a female otter originally born or captured in Minnesota will go into heat later in the winter, even if she is currently housed in Florida. Zookeepers at the Pueblo Zoo recognized the likelihood that their older female (Thelma), from Wisconsin, did not seem receptive when the males, both from Louisiana, were most interested in her. Instead, she seemed to be receptive a few weeks later, when the males were not as interested. A post-partum estrous is also possible, allowing a female NARO to give birth to one litter and immediately become pregnant with the next season s litter. There can be one to six pups in a litter, although two to three is most common (Liers 1951; Reed-Smith 2012a). Summary Captive NAROs require a nutritionally complete daily diet. Supplements, such as thiamin E paste, can help ensure they receive the necessary nutrients. Enrichment items 38

50 can also supplement the daily diet, and institutions may improve captive life with new toys and activities. At least one den should be provided for the female NAROs that is secluded from the exhibit and the male otters. NAROs breed once a year and may breed with multiple individuals. To correspond with periods of peak receptivity, to mimic wild conditions, and to avoid a confrontation, male NAROs in captivity can be rotated on and off-exhibit with the female. Providing a more natural environment will help ensure that captive NAROs lead a healthy life. Resources and Breeding Recommendations Introduction Successful captive breeding is important to the captive NARO population, because there are a limited number of new NAROs that enter the population (Association of Zoos and Aquariums 2013b). Some wild NAROs are introduced to the captive population every year, but these are mainly rehabilitation and nuisance animals (Association of Zoos and Aquariums 2013b). Successful captive breeding is also important to maintain the genetic diversity and fitness of the captive population. The Association of Zoos and Aquariums (AZA) plays a vital role in captive breeding and endangered species conservation through programs aimed at public education, professional training, research, and in situ conservation (Snyder et al. 1996). For accredited institutions, they are the governing body for animal care and husbandry. The AZA creates animal care manuals (ACM) for every species they house, which contain guidelines to provide the best possible daily care. The AZA is a forerunner in animal conservation and captive breeding. They have created a Species Survival Plan 39

51 (SSP) and studbook for most species that are bred in captivity. These manuals detail the most recent information on animal care and best practices for successful captive breeding. Association of Zoos and Aquariums The AZA is a non-profit organization that promotes programs of conservation, education, science, research, and recreation at public zoos and aquariums (Association of Zoos and Aquariums 2009). The AZA institutions are personally involved in the conservation of many different species, and take great pride in offering a high level of animal care. The AZA institutions also provide formal training for teachers and support school field trips for students. Many scientists associated with the AZA, either by their contributions to animal care and husbandry or their work with various institutions, contribute research and publish in numerous journals. Finally, the AZA institutions provide the opportunity for the general public to get involved by volunteering (Association of Zoos and Aquariums 2009). The AZA has many important guidelines for the care and preservation of wild and captive animals. The philosophy and mission of the AZA is to provide its members the services, high standards and best practices needed to be leaders and innovators in animal care, wildlife conservation and science, conservation education, the guest experience, and community engagement (Association of Zoos and Aquariums 2009). The AZA programs include zoo and aquarium field conservation, reintroduction programs, sustainable practices for zoos and aquariums, partnerships with national and international conservation efforts, many areas of research, education programs for the public, and the AZA Species Survival Plan (SSP) Program. The AZA strives to provide continued 40

52 education about and conservation of many of the world s species, whether or not they are endangered. The SSP programs create guidelines and a clear situational framework to increase the efficacy of captive breeding programs. Many educational opportunities are available to employees, teachers, students, and the general public to learn about the natural history, ecological niche, and species description of all captive species, as well as many wild animals that may not be held in captivity. In situ conservation programs increase the awareness of the needs of wild animals; and captive breeding can be an important part of both public education and reintroduction programs (Entwistle and Dunstone 2000; Association of Zoos and Aquariums 2009). The AZA strives to set high standards that will create a common ground between human needs and conservation needs while engaging the public, in effect creating a bridge between research, conservation and public life (Coe 1987). Any zoological institution in the world may become a part of the AZA organization. The AZA website (2009) states that the inquiring institution must meet the high standards that have been set to accomplish the AZA organization goals. In order to be accredited, the institution must be researched, inspected, and approved by a group of experts who determine if the institution meets the high standards of the AZA organization. Being an accredited institution gives official recognition, increases public trust, and gives access to many AZA programs and services, such as animal exchanges (Association of Zoos and Aquariums 2009). There are currently 222 accredited institutions in seven countries, including the United States, Canada, Mexico, Bermuda, Argentina, Hong Kong, and the Bahamas (Association of Zoos and Aquariums 2009). 41

53 AZA does not just benefit the accredited institutions. According to the AZA website, 175 million people visit member zoos and aquariums each year. Visits to zoos and aquariums enhance the visitors knowledge of wild animals and, more importantly, their understanding of conservation programs (Falk et al. 2007). These institutions allow visitors to put the conservation of wild animals into a personal context, which increases the likelihood that the public will become involved. This is very important in animal conservation, which ideally involves everyone. AZA role in animal care and husbandry The AZA takes their roles in animal care and husbandry very seriously. The AZA has created a hierarchy of groups who work together and with experts to create guidelines and manuals for animal care and husbandry. Some of the most important documents they create are the animal care manuals (ACMs), Species Survival Plan (SSP), and studbooks. A management hierarchy of the groups responsible for the creation and implementation of these documents is shown below (Fig. 10). The creation and management of these important documents usually involves several different groups, all of which include experts in animal welfare and members of the AZA community. This process ensures that each species will be well-represented and that their captive care will be standardized and of the highest quality at all AZA institutions. 42

54 Figure 10. Management hierarchy of the Association of Zoos and Aquariums showing the groups and committees responsible for the creation (solid arrows) and management (dashed arrows) of important animal welfare documents: Species Survival Plan, Studbook, and Animal Care Manual (adapted from Association of Zoos and Aquariums 2012). The Taxon Advisory Group (TAG) actually creates the ACMs, SSPs, and studbooks (Fig. 10). The mission of the TAG is to examine the conservation needs of an entire taxa, and to develop recommendations for population management and conservation based upon the needs of the species (Association of Zoos and Aquariums 2009). Each TAG is composed of several advisors who are experts in their respective fields: conservation, education, population, research, scientific, or technical. These advisors work together to incorporate all the needs of a species into a document created specifically for that species. The TAG works closely with the Population Management Center (PMC) to coordinate and utilize these important documents. The PMC is responsible for 43

55 maintaining and coordinating the use of these policies compiled by the TAG, and for implementing breeding and transfer plans, as necessary. Animal Care Manual (ACM) The ACMs are created by the TAG in conjunction with an animal welfare committee. These manuals provide a compilation of basic animal requirements, best practices, and animal care recommendations for each species or closely related group of species (Association of Zoos and Aquariums 2012). ACMs are regularly updated to include new knowledge gained from the latest scientific research. ACMs include a variety of categories and components that are presented in a way that allows someone unfamiliar with the species to better understand the organism and the specific recommendations for its captive housing. The categories include taxonomic information, ambient environment recommendations, habitat design and containment, transport, social environment, nutrition, veterinary care, reproduction, behavior management, animals used for education programs, and research (Association of Zoos and Aquariums 2012). ACMs function as the primary source material for maintaining an animal in captivity. Species Survival Plan (SSP) Originally, the Species Survival Plan (SSP) Program was implemented by the AZA as a way to manage endangered or threatened species. Now SSP programs include most breeding species within AZA and associated institutions. There are more than 500 SSP programs, each managed by their corresponding TAG (Association of Zoos and Aquariums 2012). For most species, all members of the population within AZA institutions are treated as a single population for breeding purposes. The Population Analysis & Breeding and Transfer Plan includes a breed or do not breed 44

56 recommendation for every animal within the population, based on the population needs and the aim of increasing genetic diversity. The breeding recommendations are used to maintain a demographically stable and genetically diverse population (Association of Zoos and Aquariums 2012). The Population Analysis & Breeding and Transfer Plan discusses the basis of genetic diversity requirements for the population, and gives the minimal requirements for achieving that goal, for example by maximizing the effective population size and maintaining equal founder representation (Frankham et al. 1986). Ideally a population would aim for 100% genetic diversity in the population over 100 years (Association of Zoos and Aquariums 2012). This means the ideal population would have enough unrelated mating individuals to produce a new generation of unrelated individuals, thus continuing a healthy, genetically diverse population. 100% genetic diversity is not always attainable, but it is useful as a goal to maintain the highest level of genetic diversity possible. Studbook A studbook is a document and computer database of pedigrees, along with the life history of each individual in a population (Frankham et al. 2007; Association of Zoos and Aquariums 2009). These documents will eventually be created for every breeding species in AZA institutions. Studbooks are used to track and manage each individual and are key to supporting captive breeding. The studbooks are used to create pedigrees for each individual. These pedigrees are used when choosing individuals for captive breeding pairings (Frankham et al. 2007), as they show the lineage and relatedness of each individual. The studbook is also used to assess the population status, determining whether it is stable, increasing, or decreasing (Association of Zoos and Aquariums 2009). A stale 45

57 population has a relatively equal number of deaths and births. An increasing population has more animals being introduced to the population, through births or wild caught, than are leaving, usually through death. Conversely, a decreasing population has more animals leaving than are coming into the population. Captive Breeding in AZA Institutions The ACM, SSP Programs, and studbooks play a vital role in captive breeding. They possess the basic demographic information regarding captive species, and give specific recommendations for their care and captive breeding. Captive breeding may aid in increasing the number of wild individuals through reintroduction programs, and it is a vital part of supplementing and maintaining captive populations. In general, animals are not taken out of the wild to increase captive populations. For NAROs, only rehabilitation and nuisance wild animals are added to the captive population (Association of Zoos and Aquariums 2013b). Thus, captive breeding allows the number of individuals in the population to increase without unnecessarily reducing wild populations. Once a captive breeding program has been established, it becomes easier to maintain that population. There is less stress on any individual institution to force breeding, and through such management it is easier to maintain a healthy and genetically diverse population across multiple institutions. Captive breeding is also important to AZA institutions because it brings awareness of animal conservation to the public. It is well-known that patrons tend to visit mammals and other entertaining animals more often (Balmford 2000). The more attention the institution can draw to its animals and programs, the more money they are likely to bring in to support their programs overall (Entwistle and Dunstone 2000). Mammals are 46

58 especially well received and gain a disproportionate amount of support and attention where conservation and money are concerned (Entwistle et al. 2000). NAROs are an important feature species at many institutions (Foster-Turley n.d. b); they often draw patrons to the zoological institutions due to their animated and playful nature. The ability of these animals to cause an emotional reaction in patrons is one reason they are featured in captivity. These displays can also highlight conservation efforts in place to preserve them in the wild (Entwistle et al. 2000; Foster-Turley n.d. b). As mentioned previously, when managing captive breeding programs, pedigrees are created and consulted to establish breeding pairs (Frankham et al. 2007). This is done to maintain and increase the genetic diversity of the population, which can be diminished by breeding closely related individuals in a small breeding population. A successful captive breeding program ultimately results in a viable and self-sustaining population (Williams and Hoffman 2009). Despite AZA s best efforts, captive breeding does not always result in consistent reproduction, and has proven difficult for many species (Snyder et al. 1996) as noted with sea otters (Larson et al. 2003). The reasons captive breeding is unsuccessful are often not well understood from the population through the individual level. Part of the problem may be the type of population available for captive breeding. These populations are typically made up of a small number of individuals and there are some inherent issues and risks when breeding small populations. The small population numbers generally have very few founders individuals at the top of the pedigree with no known genetic relationship to any other individual except their own offspring (Lacy 1989; Lacy 1994) and an initial bottleneck to the population 47

59 causes inbreeding depression and a subsequent loss of genetic diversity (Ralls and Ballou 1986; Bryant et al. 1999). Such population bottlenecks that result from a reduction in population numbers have also been shown to reduce evolutionary potential where the new population has a reduced ability to adapt and respond to changing environments (Frankham et al. 1999). Thus, inbreeding depression, caused by low population numbers of closely related individuals, lowers the overall fitness of the population and has been well-documented to correspond to an increase in juvenile mortality, among other issues (Ralls and Ballou 1986; Jiménez et al. 1994; Backus et al. 1995). In addition to inbreeding depression, years of research on captive breeding programs has shown a carry-over effect of domestication traits that are inadvertently selected during breeding (Araki et al. 2007, 2009). That is, the captive breeding environment has its own set of selective pressures that are vastly different from a wild environment (McPhee 2003). This adaptation to captivity selects for the genotypes best suited to the captive environment (e.g. tameness), which may be deleterious in nature (Allendorf 1993; Lynch & O Hely 2001). These selective pressures may also reduce subsequent reproductive capabilities (Araki et al. 2007, 2009). This is more of a problem when using captive bred individuals for reintroductions, but the possible genetic changes should be taken into consideration for wholly captive populations as well. The problems associated with captive breeding may be alleviated by rapid population growth, maintenance of large populations, regularly changing the breeding pairs to increase genetic diversity, introduction of new breeding individuals, and using mean kinship to avoid inbreeding depression (Montgomery et al. 1997; Balmford 2000; Bryant et al. 2009). Rapid growth of the population has a noticeable impact on the 48

60 bottleneck effects typically found after a founder event (in this case, the start of captive breeding) (Balmford 2000). The growth of the population increases the number of breeding individuals, which lessens the likelihood of siblings mating. Depending on the severity of the inbreeding depression, maintaining large populations typically provides a larger genetic base to increase diversity. In addition, immigration of wild animals into a captive population has been shown to minimize maladaptation to captivity, and can provide a new source of genetic diversity (Allendorf 1993; Williams and Hoffman 2009). Regularly changing the breeding pairs increases the genetic diversity by providing new breeding individuals with a different relationship to the local population. To maintain a high level of genetic diversity, captive breeding programs in the AZA focus on minimizing mean kinship (MK). MK is a number representation of the relatedness of individuals in a population (Lacy 1994; Frankham et al. 2007). MK is important for its ability to maximize the retention of gene diversity in populations with unequal founder representation (i.e. when most individuals come from a single lineage, and, therefore, provide very similar genetic information to the population diversity) (Montgomery et al. 1997). The Population Analysis & Breeding and Transfer Plans include tables showing the MK of all individuals in the population (Association of Zoos and Aquariums 2013b). MK is one consideration when choosing breeding pairs to preserve genetic diversity and reduce inbreeding (Williams and Hoffman 2009). There can be many problems associated with captive breeding programs, but some of these can be alleviated or at least reduced with carefully designed breeding plans (Ralls and Ballou 1986). The most-fit individuals should be chosen for captive breeding (Backus et al. 1995), because inbreeding is unavoidable in small populations (Frankham 49

61 et al. 2007). The most-fit individuals are those who are better able to survive in their environment and are more likely to be reproductively successful; they are, therefore, more likely to pass on the genetic material that is beneficial for that area. The most-fit individuals would be preferred for a captive breeding program because they are more likely to lead to a sustainable population (Backus et al. 1995). The Population Analysis & Breeding and Transfer Plans and the studbooks play a very important role in creating and managing a successful captive breeding population. Resources and management of NARO captive breeding NAROs are often unsuccessful captive breeders. In addition, the captive population is based on a relatively low number of founders, i.e., individuals obtained from a source population (often the wild) that have no known relationship to any individuals in the derived population (except for its own descendents) (Association of Zoos and Aquariums 2013b). This low founder number has led to a reduction of the genetic diversity that would ultimately be desired (Foster-Turley n.d.b). With increased oversight of captive breeding programs, the AZA has slowly been changing the composition of the captive population, and there has been some turn around. The initial low number of founders has been increased due to the available rehab and trapped nuisance animals that are added to the population yearly. As of 15 January 2013, the captive AZA starting population has 56 founders out of 191 potentially breeding individuals, and the total gene diversity is 97.43% (Association of Zoos and Aquariums 2013b). There is still a need for oversight of breeding and a need to understand what increases the success of captive breeding programs. To manage these issues, the AZA 50

62 uses all of its breeding resources, the ACM, SSP program, and studbook to supervise the captive population. The NARO studbook includes a life history of all NAROs in the current population. This information is used in conjunction with the Population Analysis & Breeding and Transfer Plan to determine which NAROs will breed, with whom they will breed, and which institutions will house these breeding pairs (Association of Zoos and Aquariums 2013b). Many individuals are given a nonbreeding designation and removed from breeding zoos due to age, health issues, or because they have a high MK. The AZA keeps as much control as possible over the population to ensure the genetic diversity continues to be maximized. The NARO captive breeding program is managed as a Green SSP program. A Green SSP designation meets three requirements: the population size at the time of planning (starting population) includes more than 50 individuals, is expected to retain more than 90% genetic diversity for 100 years or 10 generations, and the present population is demographically sustainable with a positive growth rate that will allow the population to be maintained for 100 years or 10 generations (Association of Zoos and Aquariums 2012). As a Green SSP, participation in all recommended practices and breeding decisions is required by all AZA institutions to maintain the NARO population. There are also Yellow and Red SSP programs for AZA captive breeding. A Yellow SSP has a starting overall population of more than 50 individuals, and the projected gene diversity after 100 years is less than 90%. These populations are potentially sustainable but will require extra attention and efforts to increase their sustainability. With increased efforts, it is possible for these populations to change to a 51

63 green designation, if their projected diversity changes to more than 90% after 100 years (Association of Zoos and Aquariums 2012). The Black-footed ferret is designated a yellow SSP (Association of Zoos and Aquariums 2013a). Red SSP programs start with an overall population of fewer than 50 individuals and they are expected to have less than 90% genetic diversity after 100 years. These populations are currently unsustainable and in critical need of new, unrelated individuals to increase their sustainability (Association of Zoos and Aquariums 2012). It is highly unlikely that these Red SSPs programs will lead to a sustainable population long term. The most recent Population Analysis & Breeding and Transfer Plan (Association of Zoos and Aquariums 2013b) -- a requirement of SSP programs -- indicates that the current NARO population is projected to maintain 90% genetic diversity over the next 32 years, but will fall to only 77% genetic diversity after 100 years. To maximize diversity, more individuals should be added, either through captive breeding or wild recruitments (Association of Zoos and Aquariums 2013b). Successful breeding of the 79 unrepresented potential founders (i.e., individuals who have not yet contributed their genetic material to the population) already in the population would also increase the genetic diversity (Association of Zoos and Aquariums 2013b). The AZA also provides optional breeding strategies to increase the chance of successful captive breeding. One example of this, detailed in the Population Analysis & Breeding and Transfer Plan, is the seasonal separation breeding strategy. The published instructions for seasonal separation (direct the institutions to separate the males and females approximately two months before the breeding season starts Association of Zoos and Aquariums 2013b). Separation prevents direct olfactory exposure, while rotating the 52

64 NAROs through the exhibit at least daily allows them to continuously smell the scent of the other individuals (Association of Zoos and Aquariums 2013b). This strategy is thought to mimic wild conditions, where males and females do not live together yearround, and possibly reduces the chances of males and females evolving a sibling bond to the point they will no longer breed due to familiarity. Summary The proper care of captive animals is the highest priority for the AZA institutions. Groups of experts work together to create management strategies and guidelines for best practices that are available to all AZA participating institutions. Only after the proper care of animals has been established does captive breeding become a priority for the AZA. Captive breeding is highly regulated and, in most cases, is essential to maintaining the captive population. ACMs, Population Analysis & Breeding and Transfer Plans, and studbooks are used to create and manage breeding plans. The breeding plans are created exclusively for each species, and the population is managed as a whole, rather than as individuals at separate AZA institutions. Many steps are taken to guarantee healthy, genetically diverse populations exist and will continue to thrive. This ensures a healthy population will continue for the enjoyment of patrons and to aid in the conservation of a variety of species. Statement of the Problem and Justification for the Internship Research Project This internship research project was designed to investigate potential reasons that captive breeding of NAROs at the Pueblo Zoo had been unsuccessful. At the beginning of the research project, the Pueblo Zoo housed two male NAROs and one female NARO. The males were wild-caught in Louisiana, and were approximately 11 years old. The 53

65 female NARO was wild-caught in Michigan and was approximately nine years old. All three NAROs have been together for more than five years, and each is biologically capable of successful breeding. Each breeding season, they have shown normal breeding behaviors, such as interest in each other, and attempted copulations, but none led to successful reproduction. The Pueblo Zoo has followed all guidelines and recommendations set forth by the AZA (Henry et al. 2012; Reed-Smith 2012a, 2012b, 2012c). For the past few years, the Pueblo Zoo has tried seasonal separation to increase the chances of successful breeding. The male NAROs were removed from the female NARO beginning in November. Once the female showed signs of estrous, one male at a time was reintroduced to her. The males were rotated to be with the female every two days. A 2010 breeding observation study (Appendix A) showed that there was normal copulation behavior between the female and both males. Knowing that these NAROs are biologically capable of reproduction, it is imperative to understand what is inhibiting their ability to breed successfully. When discussing the issue with the Curator (Marilyn McBirney) and the Primary Otter Keeper (Melanie Pococke) at the Pueblo Zoo, two categories of factors that affect captive breeding were identified that may have had an impact on successful captive breeding: 1) geographic origin of the NAROs; and 2) zoological practices. The latter category may be subcategorized into the following components: a) diet; b) den and nesting materials; c) presence of males outside of the breeding season; and d) keeper involvement. These categories became the focus of the study. 54

66 This is an important internship topic for several reasons. First, NAROs are an important feature species at zoos and aquariums, and their continued successful captive breeding will help ensure a healthy population that brings in more patrons. Second, captive breeding of the NARO has had limited success at other AZA institutions as well, and the specific factors that contribute to that success are unknown and have not been well studied. Third, because otters are rarely taken out of the wild to increase captive populations, it is essential to improve the rate of successful captive breeding. Finally, the methods used in the current study to identify factors that contribute to successful captive breeding of NAROs could have implications for reintroduction efforts for NAROs, as well as other species who have had limited success with captive breeding. 55

67 Hypotheses and Specific Aims Geographic Origins HYPOTHESIS 1: Successful captive breeding is dependent upon having two North American river otters from similar geographic locations. SPECIFIC AIM 1a: Determine whether the total distance of origination between the male and female NAROs from a mating pair affects successful captive breeding. SPECIFIC AIM 1b: Determine whether the difference in latitude of origination between the male and female NAROs of a mating pair affects successful captive breeding. Zoological Practices HYPOTHESIS 2: Successful captive breeding is dependent upon zoological practices such as the diet, the den, the housing arrangements, and the keeper involvement, which affect the daily life of captive North American river otters. SPECIFIC AIM 2a: Determine whether the daily diet, enrichment, and daily supplements are factors in successful captive breeding. SUB-AIM a-1: Determine whether the type of foods given for the daily diet affects successful captive breeding. SUB-AIM a-2: Determine whether the type of enrichment foods given affect successful breeding. SUB-AIM a-3: Determine whether vitamin supplementation affects successful captive breeding. SPECIFIC AIM 2b: Determine whether the den and nesting materials provided to captive NAROs are factors in successful captive breeding. 56

68 SUB-AIM b-1: Determine whether the location of the den (indoors or outdoors) is a factor in successful captive breeding. SUB-AIM b-2: Determine whether the type of den provided is a factor in successful captive breeding. SUB-AIM b-3: Determine whether the type of nesting materials provided is a factor in successful captive breeding. SUB-AIM b-4: Determine whether the availability of nesting materials is a factor in successful captive breeding. SPECIFIC AIM 2c: Determine whether the presence of the male NAROs with the female NAROs outside of the breeding season is a factor in successful captive breeding. SPECIFIC AIM 2d: Determine whether the amount of time keepers spend with their captive NAROs affects successful captive breeding. 57

69 Study Design Internship and Observations To gather information on potential factors that contribute to the successful captive breeding of the North American river otter (NARO), an internship was completed at the Pueblo Zoo in Pueblo, Colorado. This internship provided an opportunity to observe and work with all of the different animals housed at the zoo, with specific time dedicated to the study of the zoo s captive NAROs. The observations that were made were necessary to understand captive NARO behavior, both during and outside of their breeding season. In addition, the experiences with the NAROs, and other zoo animals, gave a better understanding of zoological practices and how zoos operate to provide the best possible captive life for animals housed there. The NARO observations were also essential to the creation of hypotheses about the factors that may contribute to their successful breeding. These hypotheses were then explored using a survey of zoological institutions. Specific internship learning goals were set with the help of Marilyn McBirney, Curator of the Pueblo Zoo, when the internship began. These goals included learning about the Association of Zoos and Aquariums (AZA) industry standards, philosophy, and programs; national and international management of endangered and threatened species in captivity; species natural history, ethology, and ecological niche; and in situ conservation programs. These learning goals are addressed more fully in the background section titled Resources and Breeding Recommendations. The ethological study of the NAROs is discussed in detail below. The learning goals were designed to provide a better understanding of the environment provided to captive animals and the reason that these 58

70 institutions exist. It was also important to understand information specific to NAROs in order to create the survey, and to make working with the otters more meaningful. Approximately 200 hours were spent at the Pueblo Zoo during the internship. Notes from the internship hours can be found in Appendix F. The internship hours were spent assisting the keepers, the veterinarian technician, and the veterinarian; providing behavioral enrichment; and participating in animal training for medical purposes. More importantly, observation skills of animal behavior and how to record this information were learned. These skills are helpful in carrying out the care, nutrition, handling, and breeding of the animals. The observation skills learned during the internship hours were used to observe the river otters throughout the year. A specific ethological study of NARO mating behaviors during the 2010 breeding season was completed (see Appendix A for complete observation notes). Understanding the behavioral differences during and outside of the breeding season was important for understanding the Pueblo Zoo NARO breeding issues. The internship hours spent with the otters created a knowledge base from which survey questions about breeding practices were created. The survey was used to gather information regarding the NARO breeding programs at other AZA institutions. As previously discussed, the Pueblo Zoo NAROs have been together for more than five years and are all biologically capable of successful breeding, although they have never bred successfully. The otters do, however, show interest in each other and demonstrate normal mating behavior each year. The question becomes, what is prohibiting successful captive breeding of these healthy, reproductive individuals? This is especially important to understand considering the success of a new female that was 59

71 added to the exhibit in Although she was only a year old at the time, she successfully bred with one of the males at the Pueblo Zoo and gave birth to a litter of three pups in spring Ethological Study An ethological study of the otters during their mating season was conducted from late January 2010 through early March Observations of the NAROs were made for two consecutive hours several days a week, switching days of the week and time of day to vary the observational situations. During these observations, the NAROs mating behaviors, the time at which these behaviors occurred, as well as the duration of these activities were noted. Individual behaviors, as well as interactions between otters, were recorded. Notes on these observations can be found in Appendix A. Most of the observations were made from the otter viewing area of the otters on-exhibit enclosure (Fig. 11). Access was given to their off-exhibit enclosure for some of the observations. The differences in regular behavior, as well as the mating behaviors, seen in these two enclosures were recorded during the study. The observations from the internship and ethological study were used to understand how the Pueblo Zoo cares for their NAROs and to propose hypotheses about what might then be influencing breeding success. This information was also used to create a survey that was sent to other institutions to gather information about the otters they house, and the zoological practices they use to care for their NAROs and their NARO breeding programs. 60

72 Figure 11. A schematic drawing from 2003 of the North American river otter exhibit at the Pueblo Zoo which shows the on-exhibit area (upper left), inside holding areas (top middle), and the offexhibit area ( otter holding pen upper right). Observations were made from the otter viewing area (lower left) and outside of the off-exhibit area ( otter holding pen upper right). Reproduced with permission of the Pueblo Zoo. Geographic origins Since the wild caught NAROs that reside at the Pueblo Zoo in 2010 originate from different parts of their natural range, and there has never been successful breeding, the Pueblo Zoo has questioned whether or not these otters will ever be successful breeders. This speculation led to the first hypothesis -- that having two otters from similar geographic locations is necessary for successful reproduction. To explore this hypothesis, the survey asked specific questions about the otters age, place of origin, and whether they were wild caught or captive bred. Zoological practices Daily diet, enrichment, and supplements: Working at the Pueblo Zoo provided an opportunity to observe every aspect of animal caretaking. It was hypothesized by keepers and by the author, via direct observation during the internship hours, that successful breeding may be dependent on more than just the origin of the otters, that is, it 61

73 may also depend on how the zoos care for them. Throughout the observation period, the types of foods, enrichment items, and supplements that were regularly given to the NAROs were noted. A nutritionally complete daily diet is essential to the overall health of every animal. To study the effects of the daily diet on successful captive breeding, respondents were asked to describe the daily otter diet on the survey. The full survey can be found in Appendix C. Different types of foods, smells, and toys are provided as enrichment to engage the otters. Every zoo offers different types of enrichment foods, thus respondents were asked on the survey to, please describe enrichment foods. This question corresponds to the second hypothesis that diet, enrichment, and supplements are factors in successful captive breeding. There are many types of nutritional supplements available for NAROs; the survey asked the responding institutions to list their preferred daily supplementation. It was important to study the supplements in case they play a role in successful captive breeding. Den and nesting material: A diagram of the inside rooms of the Pueblo Zoo otter exhibit is shown in Figure 12. OH stands for otter holding and refers to the separate spaces available. 62

74 Figure 12. Diagram of the indoor North American river otter holding (OH) areas. OH 1 offers a plastic box (~30 x20 x15 ) as a den for the female otter. Adapted and reproduced with permission from a schematic drawing created by the Pueblo Zoo in OH 1 is a reserved space for the female otters only. It includes a plastic nest box (approximately 30 x 20 x 15 ) for females to use as a den during the breeding and pupping seasons. Pueblo Zoo provides different nesting materials once the breeding season begins. The type of den and the type of nesting materials provided for females, in anticipation of a pregnancy, were noted. These observations led to the third hypothesis that the den and nesting materials are factors in successful captive breeding. The survey asked the institutions to describe the type of den available to females and whether the den is a pen enclosure or diggable area. The type of nesting materials given and whether they are available seasonally or year-round was also surveyed. 63

75 Presence of male otters outside of the breeding season In the wild, male and female otters rarely spend time together outside of the breeding season (Hancox 1992; Larivière & Walton 1998). Because the Pueblo Zoo houses their males and females together year-round, a question was included on the survey asking the responding institutions if they housed their males and females together or separately. Due to the difficulties experienced trying to breed the otters, the Pueblo Zoo now practices seasonal separation. The guidelines for seasonal separation are outlined in the North American river otter Species Survival Plan (Association of Zoos and Aquariums 2012) and discussed in the Resources and Breeding Recommendations section. This strategy requires the males and females to be separated two months prior to the typical breeding season. At the Pueblo Zoo, the two males are rotated through the exhibit after the female is present so the males can continuously be around the female s scent. Once the female is in estrous, the males are introduced to the female one at a time, and rotated regularly. This more closely mimics natural conditions where males and females only meet during the breeding season (Hancox 1992; Larivière & Walton During the breeding season, the males were removed from the female in December, to comply with seasonal separation protocols. The males were individually reintroduced again in February. Once a male was introduced, the pair was together for two days, and then the males were switched. This was done to mimic nature and to reduce possible fighting between the males. During the ethological study, the days and times of observation were varied to see how the NARO relationships evolved and differed. 64

76 Keeper involvement: The last questions of the survey relate to the keepers involvement with their NAROS. Specifically, the survey included a question pertaining to the amount of time the keepers spent around their NAROs each day. Also questioned were the possible effects of the keepers on normal breeding behavior. The Pueblo keepers are very aware that when Thelma, the older female, is familiar with someone, she will often swim circles in the front windows whenever they are within eyesight. This behavior, and her disinterest in all other activities during her circling, had already been noted by the keepers. It is speculated that this is one reason the regular staff had not seen Thelma copulate, while patrons had. Thelma was also hand raised, so her focus was often on the keepers. The last question of the survey asks if the respondent feels that the keepers have an impact on otter reproduction. The option was given to explain the answer. Summary The hours spent during the internship were essential to understanding the NARO captive breeding problem at the Pueblo Zoo. The work that is required to provide a healthy environment for all captive animals was observed. The efforts to create a natural environment that will promote captive breeding are immense, and involve all aspects of daily care, because healthy animals are required for successful captive breeding. The hypotheses explored in this study were created from direct observations working with the NAROs, in conjunction with the zookeepers. Hypotheses were addressed via a survey of other zoos and aquariums to gather more information about captive NARO breeding. These data were compared to understand the effects of the 65

77 origins of otters, as well as zoological practices as factors that lead to successful captive breeding. Data Collection It was determined that the most straight-forward way to gather information relating to factors that contribute to successful captive breeding of NAROs was to ask specific questions in a survey directed to the people who care for them regularly. The survey was created with the professional input of Marilyn McBirney, Curator of the Pueblo Zoo, and Melanie Pococke, Primary Otter Keeper of the Pueblo Zoo. The observations made during the internship hours were used to determine the basic categories of questions. These categories included otter details, geography, history of successful breeding, housing, diet and enrichment, exhibit, den/nest box, and keeper involvement. Specific questions were asked within each category to gather the necessary information for analysis. Every question on the survey was agreed upon by the author, in consultation with McBirney and Pococke. Each question was created to gather specific information to explore each hypothesis. After compiling the survey questions, the survey was revised and approved by David Hamilton, NARO studbook keeper and NARO SSP coordinator at the Seneca Park Zoo, Rochester, NY. The survey was posted through Survey Monkey, an online survey site, to make data collection easier. In June 2011, David Hamilton ed a link to the survey using the NARO Institutional Representative (NAROIR) list, which includes 133 AZA institutions that currently house NAROs. This included a personal request from the author briefly describing the internship project and asking the 66

78 zoo representatives to respond to the survey inquiries to the best of their abilities. A copy of both the message and survey are included in Appendix C. Even after careful checking, there were a few issues with the first survey, and some institutions had trouble viewing and completing it. Due to these issues, a new survey with an improved, more user-friendly format was created. This second survey, which asked for all of the same information as the first, was ed by David Hamilton about two weeks after the first , through the same NAROIR list. Any institutions that had started the first survey were contacted directly by with the new link. A short apology for the inconvenience was included. In the beginning of July 2011, David Hamilton sent out a reminder request through the NAROIR list, and one month later a personal request from the author was sent, again through the NAROIR list. Marilyn McBirney was asked to identify any other institutions that had had successful captive breeding, but had not yet responded to the survey. She identified six such zoos, which were contacted directly by to fill out the survey. To increase the number of institutional responses for data analysis, an internet search for institutions that housed NAROs was completed. Using this list, 36 AZA institutions and seven non-aza institutions were called and requested to begin the survey. A slightly shorter survey was created for these new contacts. Some questions, that were determined to be unnecessary after responses to the original survey were analyzed, were deleted. For example, questions about the size and specifics of the exhibit were deleted because they provided no useful information. The purpose of shortening the survey was to increase the chances that the new contacts would complete it. At the same 67

79 time, 24 AZA institutions were ed and requested to finish a previously started survey. Appendix B contains five lists of contacted zoos according to whether they are AZA accredited successful breeders (14 (10%)), AZA accredited unsuccessful breeders (16 (11.4%)), AZA accredited non-breeders (20 (14.3%)), AZA accredited who did not respond to the survey (84 (60%)), and non-aza institutions (7 (5%)). Data Analysis All Survey Monkey responses were downloaded and put into one large excel file. Fort Worth Zoological Park and Sedgwick County Zoo had ed separate files with diet information, so this information was added to the larger file and their s were saved for future reference. The responding zoos were separated into successful and unsuccessful institutions based on whether they had achieved successful captive breeding in the past, or had attempted breeding without any success. Responding zoos which had never tried breeding their otters, or only housed a single sex, were left out of the analysis. Maps were created using the geographic distance data ( as the crow flies ) to help visually interpret the distance of origin between the male and female river otters of a mating pair, as well as to compare the difference between successful and unsuccessful pairs (Fig. 13, Fig. 14). Maps showing the potential natural geographic boundaries as well as the potential subspecies boundaries were created to visualize whether these differences might influence successful versus unsuccessful breeding. The distance of origination data was also used to create a pedigree for each mating pair. The pedigrees were then used to identify any possible trends within the distance of origination of all mating pairs, including relatives of the NAROs within the data set. The pedigrees can be found in Appendix D. 68

80 The rest of the data was visually analyzed using lists and bar graphs. Lists were used to compare the types of daily foods, enrichment items, den materials, and nesting materials. These lists were then compared to find the similarities and differences. Any items listed by both successful and unsuccessful breeding institutions were removed, because these were factors unlikely to be germane to the hypotheses. The items that remained were then explored to identify any possible trends. Bar graphs were created to compare the types of daily supplements, location of dens, types of den, seasonality of nesting material, and amount of interaction between keepers and otters. The bar graphs were analyzed for trends before performing a student s t-test, if possible. The sample size (n), mean, and standard error of the mean (using a 90% or 95% confidence interval) were generated. No additional statistical analysis was feasible due to the nature of the data. As this was an observational study rather than an experiment, there were no control groups nor were there any treatments groups. It was also not possible to perform t-tests on the specific aims related to zoological practices due to the inability to generate numerical means. 69

81 Results One hundred and forty-one different zoos and aquariums, of which eight were non-association of Zoos and Aquariums (AZA) institutions, were contacted by and/or phone (Appendix B) to complete the North American river otter (NARO) survey. Thirty institutions completed the survey, giving a 21.3% completion rate. Of those thirty, 15 were successful captive breeding institutions (Pueblo Zoo after the addition of a new female in 2013) and 16 were unsuccessful captive breeding institutions (Pueblo Zoo before the addition of the new female). Not all institutions responded to every question on the survey. Geographic Origins The map results for distance of origination of the male and female river otters of a breeding pair illustrate that having two NAROs from similar geographic locations and originating within a shorter distance from each other aids in successful captive breeding (Fig. 13, Fig. 14). The maps also show that a greater number of unsuccessful breeding pairs have male and female otters that cross putative subspecies boundaries. 70

82 Figure 13. Map of the United States of America comparing the distances of origination of male and female North American river otters of successful breeding pairs with regard to natural geographic boundaries and subspecies boundaries. Locations were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in

83 Figure 14. Map of the United States of America comparing the distances of origination of male and female North American river otters of unsuccessful breeding pairs with regard to natural geographic boundaries and subspecies boundaries. Locations were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Comparing the two maps illustrates that unsuccessful breeding pairs have on average a larger distance between the origin of males and females compared with the origination 72

84 Average distance of origination of male and female river otters in a breeding pair (km) distance between successful breeding pairs. There is also a trend in the number of unsuccessful breeding pairs whose male and female otters would have to cross natural geographic boundaries (i.e., mountain ranges) if they were to mate in the wild. To assess these trends numerically, the average distance of origination between the male and female river otters of successful versus unsuccessful breeding pairs were compared using a one-sided t-test, the results of which are found in Figure 15, below * 0 Successful pairs Unsuccessful pairs Figure 15. Mean (± SE) of the distance of origination between male and female North American river otters of successful (n = 14) and unsuccessful (n = 17) breeding pairs were compared using a one-sided t-test. The results show a significant difference (p = 0.02 at α = 0.05). Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in The average distance of origination between successful and unsuccessful breeding pairs is statistically significant (p = 0.02) at α = To assess the possibility that two otters may be successful outside of their natural and potential subspecies boundaries as long as they come from similar latitudes, the average difference in degree latitude of the male and female otters of successful breeding 73

85 Average distance between latitude of oirgination of the male and female river otters of a breeding pair (km) pairs was compared with the average difference in latitude of the male and female river otters of unsuccessful breeding pairs. This was done using a one-sided t-test; the results of this test are found in Figure 16, below Successful pairs Unsuccessful pairs Figure 16. Mean (± SE) of the difference in latitude of origination of the male and female North American river otters of successful (n = 14) and unsuccessful (n = 17) breeding pairs were compared using a one-sided t-test. At α = 0.1, there is a nearly significant difference (p = 0.07). Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in There is a trend that shows latitude may be a factor in successful captive breeding and the average difference in degree latitude of origination was almost statistically significant (p = 0.07, α = 0.1). Zoological Practices Statistical analysis of the zoological practices data was not feasible, because there were no control groups nor were there any treatments groups. It was also not possible to perform student s t-tests on the specific aims related to zoological practices due to the 74

86 lack of numerical means for testing. Qualitative analysis was performed instead, using bar graphs and tables to identify trends. Daily diet, enrichment, and supplements There is no easily discernible difference in the types of foods given by successful and unsuccessful breeding institutions (Table 1). That being said, the foods given by successful breeding institutions only (left column) are more natural and could be found in the wild. Those give by unsuccessful breeding institutions only have some manufactured foods (dog biscuits and dog chow) that would not be found in the wild, including more commercial versus rough fish. This possible distinction between successful and unsuccessful institutions is mitigated by the fact that some institutions also reported giving different types of dry chow, including polar bear chow, which would not be found in the wild. Table 1. Foods given as part of the daily diet for North American river otters by successful breeding institutions (n = 14) are compared with foods given by unsuccessful breeding institutions (n = 16). Foods reported as given by both successful and unsuccessful institutions are shown in the far right column. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Successful daily foods Unsuccessful daily foods Daily foods consumed by both successful and unsuccessful breeding pairs a goldfish blue crab bones mealworms/crickets canned tuna dry chow (dog, cat, ferret, etc.) perch fish dog biscuits eggs salmon fish fresh produce raw meat small animals a Foods are categorized for ease of viewing. See Appendix D for a full list of foods given by each institution. The types of enrichment foods given by successful and unsuccessful institutions do not show any obvious differences that would be a factor in successful captive breeding 75

87 (Table 2). Although many different types of enrichment were reported, there were too many different types to easily compare their influence. Table 2. Enrichment foods given to North American river otters by successful breeding institutions (n = 13) are compared to enrichment foods given by unsuccessful breeding institutions (n = 15). Enrichment foods reported by both successful and unsuccessful institutions are shown in the far right column. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Enrichment foods given to both Successful enrichment foods Unsuccessful enrichment foods successful and unsuccessful breeding pairs a birds caught by NAROs cat food bones bloodsicles cheerios cheese whiz cat food smears crab dry kibble mixes (dog, cat, etc.) cored apples with fish inside earthworms fish omnivore diet fish goo fresh or cooked eggs peanut butter smears frogs fresh produce scallops jello ice blocks/treats tuna smears monkey chow insects mussels raw meat night crawlers rawhide peanuts sea food pine nuts small animals rosehips whipped cream salmon eggs seaweed superworms a Foods are categorized for ease of viewing. See Appendix D for a full list of foods given by each institution. Successful breeding as a function of daily nutritional supplements is demonstrated in Figure 17. Vitamin B1, vitamin E, and thiamin E paste (a combination of vitamin B1 and vitamin E) stood out among the reported supplements as being given more often than the others. These supplements are also specifically recommended for supplementation by the AZA husbandry manual (Reed-Smith 2012b). 76

88 Total number of breeding pairs Successful institutions 4 Unsuccessful institutions Figure 17. Comparison of the type of daily supplementation given to North American river otters at successful institutions (n = 14) and unsuccessful institutions (n = 15) focusing on those that give vitamin B1, vitamin E or thiamin E paste, no supplement, or some other type of supplement. See Appendix D for the specific types of supplements given by each institution. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in a Supplements are categorized for ease of viewing. See Appendix D for a full list of supplements given by each institution. Other None Vitamin B1, Vitamin E or Thiamin E paste Type of daily supplement a Limited data precluded statistical comparison of the means, however a trend can be observed. Nine out of 14 successful institutions gave vitamin B1, vitamin E, or thiamin E paste regularly. Den and nesting materials Each institution provides a den for their female NAROs, although there are no specific recommendations for the location or type of dens provided. Results for whether 77

89 Total number of breeding pairs the institutions provide an indoor or an outdoor den are shown in Figure 18, below. The type of den provided, pen or diggable, is displayed in Figure Successful institutions Unsuccessful institutions Indoors Outdoors Both Location of dens Figure 18. Comparing the location of dens available to the female North American river otters with successful institutions (n = 14) and unsuccessful institutions (n = 15) reveals a trend that having indoor dens aids in successful captive breeding. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Figure 18 reveals a trend that almost all successful institutions (78%) provide at least one indoor den, and in some cases both an indoor and an outdoor area. More than half of unsuccessful institutions, on the other hand, have only outdoor (20%) or both indoor and outdoor (40%) dens, suggesting that providing outdoor dens may be detrimental to successful breeding. 78

90 Total number of breeding pairs Successful institutions Unsuccessful institutions 0 Pen Diggable Area Both Type of den Figure 19. Comparison of the type of pen available to female North American river otters at successful institutions (n = 11) and unsuccessful institutions (n = 9) reveals a trend that pen style dens may aid in successful captive breeding. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Pen style dens may be a factor in successful captive breeding, as demonstrated in Figure 19. Eight out of 11 successful institutions provide a constructed pen rather than a diggable area. On the other hand, more than half of the unsuccessful institutions provide either only a diggable area (22%) or both a pen and a diggable area (33%), again suggesting that providing a diggable pen may be detrimental to successful breeding. Because there are no strict recommendations for the types of nesting materials that should be provided for pregnant females, each institution gives materials at their own discretion. Table 3 provides lists of nesting materials as reported by successful institutions, unsuccessful institutions, and the materials that were given by both successful and unsuccessful institutions. 79

91 Table 3. Comparison of the type of nesting material made available to female North American river otters at successful institutions (n = 14), unsuccessful institutions (n = 14), and materials reported by both successful and unsuccessful institutions. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in Successful nesting materials Unsuccessful nesting materials Similar nesting materials a pine shavings carwash strips blankets grass hay leaves mulch paper towels phone books prairie hay shredded paper straw timothy hay toilet paper towels wood wool/shavings a Materials are categorized for ease of viewing. See Appendix D for a full list of materials given by each institution. There are many different nesting materials given to captive breeding NAROs. Most institutions provide materials that could be found in nature or are similar to materials found in nature, as shown in Table 3. There are a few exceptions; most notably that one unsuccessful institution provides carwash strips as a nesting material. See Appendix D for a full list of materials as reported by each institution. In the wild, NAROs fill their dens with nesting materials only during the breeding season. The results of providing nesting materials seasonally or year-round are shown in Figure

92 Total number of breeding pairs Successful institutions Unsuccessful institutions 1 0 Seasonal Year-round Availability of nesting material Figure 20. Comparing the seasonal availability of nesting materials available to female North American river otters of successful institutions (n = 10) and unsuccessful institutions (n = 14) shows a slight trend that seasonal materials may favor successful captive breeding. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in The results, as shown in Figure 20, suggest that restricting the nesting materials to the breeding season may have some benefit. Seventy percent of successful breeders were provided nesting materials seasonally, while 64% of unsuccessful breeders were provided nesting materials year-round. Presence of male NAROs outside of the breeding season In the wild, male and female NAROs are only together during the breeding season. In captivity, there are many different arrangements for housing otters; each institution makes arrangements at their own discretion. Figure 21 shows a comparison of the percentages of successful and unsuccessful breeding pairs that are housed together 81

93 Total number of breeding pairs year-round or seasonally (breeding season only). One unsuccessful institution has tried both housing arrangements Year Round Seasonal Both Males and females together Successful institutions Unsuccessful institutions Figure 21. Comparison of the total number of breeding pairs for successful institutions (n = 13) and unsuccessful institutions (n = 18) where the male and female North American river otters are housed together year-round, during breeding season only, or have attempted both housing situations. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in The data show that 77% of successful institutions house their male and female NAROs together year-round versus 23% who house them together seasonally. However, the percentages for unsuccessful institutions are similar (78% year-round, 17% seasonal pairings). Some institutions with unsuccessful breeding pairs (6%) have tried to increase their success by changing the housing arrangements of their NAROs between year-round and seasonally pairing. Thus, the seasonality of male-female interaction does not appear to play an important role in breeding success. 82

94 Total number of breeding pairs Keeper involvement The amount of interaction the keepers have with their NAROs is compared between successful and unsuccessful institutions in Figure Successful institutions Unsuccessful institutions 0 0 Low Medium High Amount of daily interaction Figure 22. Comparison of the amount of daily interaction, low (< ½ hr), medium (> ½ up to 1 hr), and high (> 1 hr), between the keepers and the North American river otters at successful institutions (n = 12) and unsuccessful institutions (n = 17). Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions in There appears to be a trend in the responses with higher interaction leading to less successful captive breeding. Ten out of 12 (83%) successful institutions had only a medium amount (> ½ up to 1 hr) of interaction with their NAROs, while 47% of unsuccessful institutions had more than one hour of interaction per day with their NAROs. 83

95 Other Considerations There are several other potential factors in successful captive breeding (Appendix D), but data were highly variable, so no analysis was performed. One such factor was age. All surveyed otters were within the known breeding age range, so these data were not analyzed. 84

96 Discussion and Future Studies Although there was a low response rate to the survey for this internship project (21%), many attempts were made to gather data from additional institutions (Appendix B). While this is an internship rather than a research project, the low number of responses precluded a statistical analysis of all survey questions. Despite the low response rate, the results show that the geographic origins and perhaps the latitude of origination have a significant impact on the success of mating pairs, indicating that the male and female of a breeding pair are more likely to be successful if they come from similar locations. Latitude may also be important in choosing the male and females of a breeding pair, and warrants further study. A qualitative analysis of the zoological practices data was used to identify the AZA and IUCN/SSC Otter Specialist Group NARO care guidelines that may impact successful reproduction of North American river otters. In general, there is a fair amount of variation for the daily diet, enrichment, supplementation, den, and nesting materials. But some patterns emerged. The impact of the presence of male NAROs outside of the breeding season and the effects of keeper interaction on captive NAROs were also explored. Geographic Origins The data show that the distance of origination (Specific aim 1a) between the male and female NAROs of a breeding pair is a statistically significant factor (n = 14, p = 0.02, α = 0.05) when comparing successful and unsuccessful breeding pairs (Fig. 15). This corresponds to wild conditions where males, though they may travel long distances to find a mate, will usually stay within a specific home range, of around 155 km² (Hill 85

97 1994). Some male home ranges, however, have been observed to be up to 250 km 2 along a river (Kruuk 2006). The subspecies boundaries used to define NAROs in this study are generally accepted by the scientific community today and are used for the official AZA husbandry manual (Reed-Smith 2012 a). Over the years, the number of different subspecies, and what differentiates them, has been defined and debated by many researchers. For example, Hall and Kelson (1959) described 19 different subspecies, dividing them by major drainage systems and other isolating barriers instead of by physical differences. Both Hall (1981) & Larivière and Walton (1998) agree on the seven subspecies, based loosely on differing physical characteristics of the otters. For this project, one subspecies boundary (Fig. 6) accounts for the majority of captive breeding otters (86%), the lataxina and Canadensis subspecies. The canadensis subspecies also encompasses most of the eastern United States where the majority of NAROs live. Some breeding pairs did cross the recognized potential subspecies boundaries; however the only successful breeding pair that came from across different subspecies boundaries, originated from the same latitude (Fig. 13, Fig. 14). This may indicate that the latitude of origin may be an important subspecies designation in promoting successful breeding. The results also suggest that captive males and females who are going to become a breeding pair should originate from as similar an area as possible (within geographic boundaries, subspecies, latitude, and nearby origin) to increase the likelihood that their pairing will be successful. The difference in latitude of origination (Specific Aim 1b) of the male and female NAROs of a breeding pair is nearly significant (n = 14, p = 0.07, α = 0.1) when 86

98 comparing successful and unsuccessful breeding pairs (Fig. 16). Although this result was found with a lower confidence interval (90% instead of 95%), it is likely that the results would show an even stronger relationship between similar latitudes and successful captive breeding had there been more survey responses. The data that are available reveal that several unsuccessful breeding pairs originated from very different latitudes (Fig. 16). This same pattern is seen in many different families and species, including wolves (Canis lupus) and some birds (Accipitres and Coraciiformes), where breeding occurs earlier at lower latitudes (Baker 1939; Mech 2002). Most notably, in nature the timing of estrous is significantly different between NAROs living in northern and southern latitudes (Reed-Smith 2012a). This same phenomenon is also seen in black-footed ferrets (Mustela nigripes), another member of Mustelidae. When transported from northern to southern latitudes, ferrets will reverse their breeding season to conform to their new environment (The Duke of Bedford and Marshall 1942; Miller et al. 1996). This suggests that latitude has a significant impact on the success of breeding pairs suggest that latitude should be taken into consideration when choosing NAROs to breed in captivity. Although it seems that NAROs should be able to adapt to their new latitude and breed successfully, this has not been the case. The Center for Conservation and Research of Endangered Wildlife (CREW) at the Cincinnati Zoo and Botanical Gardens tests fecal samples of potentially pregnant female NAROs each year. They test for specific hormones that indicate whether a female is potentially pregnant, not pregnant, or has already had a litter of pups. Throughout their years of testing, they have noticed that female otters do not always respond to the environmental cues of a new 87

99 institution/latitude. They have yet to determine why timing of estrous is less flexible in NAROs than in other species, such as the black footed ferret (Center for Conservation and Research of Endangered Wildlife; Bateman 2014). Zoological Practices Although no further statistical analysis was feasible, qualitative analysis of the data was completed using bar graphs and tables to identify trends. Daily diet, enrichment, and supplements The daily diet items (Sub-Aim 2a) reported, which consisted of fish, meat products, hard items (dry chow), and produce (Table 1), all comply with the AZA and IUCN/SSC Otter Specialist Group guidelines (Henry et al. 2012; Reed-Smith 2012b). The main types of fish reported were capelin, smelt, herring, and trout. The guidelines suggested that these types of fish be fed, due to their low levels of thiaminase (Reed- Smith 2012b). Many of the items suggested by Owens et al. (2012) and Henry et al. (2012), such as produce and hard items, were also reported as part of the daily diet. The responding institutions appear to give a nutritionally diverse diet that includes at least one food from all recommended categories (Table 1, Appendix D) (Henry et al. 2012; Reed- Smith 2012b). There were a myriad of enrichment items (Sub-Aim 2b) reported (Table 2), most of which were food items. The items listed are all approved by the AZA and IUCN/SSC Otter Specialist Group guidelines (Henry et al. 2012; Reed-Smith 2012b). Although not specifically reported, there are many different means of providing the enrichment. For example, the Pueblo Zoo puts fish and other food pieces into plastic containers, which are then placed in the exhibit (Appendix F). The NAROs have to work to get the food out of the container. By providing enrichment foods in this way, the keepers have provided the 88

100 NAROs with extra food and a new toy. These are both important actions to provide for their nutritional needs and to prevent abnormal repetitive behaviors (Mason 1991). Daily supplementation (Sub-Aim 2c) varied greatly, although many institutions reported giving some form of vitamin B1, vitamin E, or thiamin E paste (Fig. 17). This corresponds to the AZA and IUCN/SSC Otter Specialist Group captive NARO recommendations (Reed-Smith 2012a). Many institutions also gave some other type of vitamin supplementation (Fig. 17, Appendix D), which may or may not be meeting the captive NARO s nutritional needs. The NARO diet has not been well studied (Henry et al. 2012), especially concerning the needs of captive animals. As previously mentioned, domestic cats (Felix catus) form the basis of the nutritional guidelines for NAROs in AZA institutions (Henry et al. 2012). That being said, responding institutions appear to be following the published guidelines, and are providing a nutritionally diverse and complete daily diet for their captive NAROs (Table 1, Table 2, Fig. 17), including enrichment items and supplementation. The data suggest that more natural foods (mealworms and crickets) and rough fish may enhance breeding success, while manufactured foods (dog chow and biscuits) may be counterproductive. Even if diet, enrichment, and supplementation are not factors in successful captive breeding in and of themselves, they are important to overall health, and improve captive living for NAROs. Den and nesting materials The results for location of dens provided (Sub-Aim 2d) (Fig. 18) indicate that all successful institutions provide indoor dens for their female NAROs. All successful institutions also provide a pen style den (Sub-Aim 2e) (Fig. 19). Having a ready-made 89

101 den that is secluded from the public and other NAROs is consistent with NAROs in the wild who do not dig their own dens and prefer secluded, protected areas (Melquist and Hornocker 1983). Female NAROs will modify an existing den to suit their needs, congruent with captive females adding nesting material to the dens provided for them (Liers 1951; Melquist and Hornocker 1983). The results also show a trend with the type of nesting material provided (Sub-Aim 2f) for captive female NAROs (Table 3). Successful institutions offer more natural materials than unsuccessful institutions. The availability of nesting materials (Sub-Aim 2g), whether seasonally or year-round, may not matter (Fig. 20), though successful institutions mostly offered these materials seasonally. In the wild, female NAROs only utilize nesting material during the breeding season when she makes her natal nest. Presence of male NAROs outside of the breeding season Contrary to wild conditions, allowing the male and female NAROs to live together year-round (Specific-Aim 2c) appears to aid successful breeding (Fig. 21). The results show that 83% of successful institutions do not separate their NAROs after the breeding season has come to an end. This is contrary to wild conditions where males and females only interact during the breeding season, and then only for brief periods of time (Liers 1951; Melquist and Hornocker 1983). Although 75% of unsuccessful institutions do house their males and females together year-round, their lack of success may be due to other factors. Some institutions have attempted a seasonal separation breeding strategy. The published instructions for seasonal separation (Association of Zoos and Aquariums 2013b) direct the institutions to separate the males and females approximately two 90

102 months before the breeding season starts. The NAROs are supposed to be rotated through the exhibit at least daily. This allows them to continuously smell the scent of the other individual (Association of Zoos and Aquariums 2013b). This strategy is thought to mimic wild conditions and reduce the chances of males and female evolving a sibling bond to the point they will no longer breed. The Pueblo Zoo has used the seasonal separation breeding strategy (Appendix A), although this strategy was not effective in creating a successful breeding environment for its female, Thelma, and two males. Keeper involvement The results indicate that a moderate amount (> ½ up to 1 hr) of interaction between the keepers and their captive NAROs (Specific Aim 2d) is not detrimental to successful captive breeding (Fig. 22). Keepers from successfully breeding institutions interacted with their NAROs no less than half an hour, and keepers from two successful institutions spent more than one hour around their NAROs. In contrast, of keepers from unsuccessfully breeding institutions, 47% spent more than one hour around their NAROs, and 20% spent less than half an hour. An important aspect to these results is how the NAROs are affected by the presence of keepers, and how the keepers presence may affect the NAROs normal routines. An example of this situation is found at the Pueblo Zoo. The staff there has noticed that all activity, breeding or otherwise, will stop when the keepers walk by the NARO exhibit. For example, the NAROs will see the keepers walk by their habitat and will immediately run over to the door that leads to their inside holding area. The NAROs seem to associate the presence of the keepers with feeding, so they prepare to be moved off-exhibit to allow the keepers to clean and distribute more food. The author recognized 91

103 a similar problem from her experiences observing the NAROs during the breeding season (Appendix A). By the end of the observations, the author had to remain out of sight so that Thelma, the older female, would stop swimming circles in the front windows, and return to her normal activities. Due to these experiences, the Pueblo Zoo keepers believe that they hinder regular captive breeding activities. A final question was added to the survey to address whether keepers thought they had an impact on. The last question asked the respondent to give their opinion on whether or not the keepers had an impact on captive breeding (Fig. 23). NAROs can be wild-caught or captive-bred. Whichever way they are acquired for captive populations, NAROs can either be raised by a natural parent, or hand-raised by humans. Thelma, the older Pueblo Zoo female, was hand raised. She is less fearful of people and more likely to equate them with food and handling than a parent-raised otter. Therefore, it is possible that parent-raised, especially wild-caught otters, would pay less attention to the keepers. 92

104 Total number of breeding pairs Successful institutions Unsuccessful institutions No Yes Other In your opinion, do the zookeepers have an impact on captive breeding? Figure 23. Responses to the question, "In your opinion, do the keepers have an impact on captive breeding? Responses from successful institutions (n = 13) and unsuccessful institutions (n = 13) were compared as to whether or not the survey respondent felt that their institutions zookeepers had any effect on the captive breeding of the NAROs. Data were collected using a survey sent to Association of Zoos and Aquariums accredited institutions The results indicate varying opinions as to whether keepers affect captive breeding. Approximately 35% of unsuccessful institutions do not believe they have an effect on captive breeding of their NAROs, compared with 23% of successful institutions who do not believe they have an impact. These numbers compare to 15% of unsuccessful institutions and 19% of successful institutions, such as the Pueblo Zoo keepers, who believe they do have an impact on breeding. The differences could result from some keepers being hyperaware of the animals throughout the day, while other keepers pay more attention to the NAROs when cleaning and feeding only. 93

105 It is important to note that many keepers feel that they do not have an impact on breeding, yet the results of time spent around NAROs as it relates to success (Fig. 22) indicate that having only a moderate amount of interaction with captive NAROs aids in successful captive breeding. This information should be considered for future research as the amount of interaction may be a relevant factor in successful captive breeding. It is also important to reeducate keepers to the possibility that they may have an impact on breeding. Perhaps the amount and type of interaction with breeding NAROs should be reevaluated to decrease potential negative impacts. Study Challenges and Improvements There were several challenges to this particular project, one of which was the length and breadth of the survey. As mentioned in the study design, there were a few technical flaws with the first survey that was sent to the NAROIR (Appendix C). After these issues came to light, a second survey was created to address the issues. The second survey was easier to understand and had a simplified way to enter responses, but it was long, with 50 questions (Appendix C). The third survey was shorter and more focused to the specific aims, but there were still many multipart questions and a significant amount of information was requested (Appendix C). Even with these improvements, it was still rather long. Another challenge of the project was the general difficulty of the subject. NAROs are living things, so there are multiple factors that may impact successful breeding. It may be difficult to fully understand and collect data, not only regarding each factor but also on how all these factors interact with and influence each other. The possibility exists 94

106 that more than one, or that, in fact, all of these factors work together to aid in successful captive breeding. Future Studies There are many future possibilities for this subject matter. If this type of study were to be repeated, the survey should be shortened and questions be very specific to studying one subject at a time. Researching multiple factors may prove to be asking for too much information. Finding a reliable way to increase the response rate would also be advisable before attempting to use a survey to gather information. Each of the subjects studied in this research project could be studied in greater depth. Diet, enrichment, and supplements could be studied in terms of the specific amounts of each nutrient (protein, fat, carbohydrates, etc.), that captive NAROs are receiving regularly. To do this, a survey could ask respondents to give exact amounts of foods that are given regularly. The amounts could then be compared between successful and unsuccessful institutions, looking for significant differences in a nutrient analysis. The den and nesting materials could be used for a simple experiment designed to test the female NARO s preferences. Participating institutions could provide a den and nesting materials that conform to the experimental design, providing different types and sizes of den. The nesting materials could be closely monitored at each institution to study the types preferred by each female, as well as the amount of nesting materials used. The results of such experiments could be used in future years to improve the chances of successful breeding, if indeed these were contributing factors. One important future study would be to assign breeding pairs based on the geographic origin and latitude of origin for the male and female of a breeding pair. These 95

107 factors are taken into consideration when assigning mating pairs at the moment, but they are not the top priority. If it is possible to only assign potential mates who originate from similar areas, the success rate of those pairings could be compared to the success rates of NAROs who originate from further away. Subspecies could be thoroughly studied by taking blood samples from each individual in the captive population and performing genetic tests to determine what differences occur at the DNA level. This would be a substantial undertaking, but could be useful to fully understand the importance of designated subspecies in breeding success. If successful captive breeding cannot be accomplished and maintained using the current guidelines, this may be a necessary step to ensure a genetically diverse population is continuing with the current breeding pairs. Genetic diversity in a stable population is the most important end product of the captive breeding program. The captive population blood results could also be compared to wild populations, if blood samples were available from wild populations as well. 96

108 Conclusions and Recommendations Based on this study, I recommend that the male and female NAROs of breeding pairs should be chosen based on their geographic origin, selecting those from similar locations. It also appears that the distance of origination may be greater, provided that the difference in the latitude of origin of the pair is small. Additionally, AZA institutions should continue to provide a nutritionally diverse and complete diet that includes enrichment items and supplementation, such as daily rough fish, hard bones for dental health, and appropriate vitamins. As for the den and nesting material, though evidence is anecdotal, I would recommend that an indoor pen or den area be provided, along with naturally-occurring nesting materials that are provided during the breeding season only. Based on the study results, male and female NAROs may continue to be housed together year-round. Additionally, keepers should limit the time they spend with their NAROs. Training is important both for medical purposes and for captive living, and therefore, should be continued. The necessary amount of time spent around the NAROs should be evaluated and kept to a minimum, paying close attention to whether keeper interaction may be interfering with breeding behavior. The Pueblo Zoo responses to the survey indicate that they are following all necessary guidelines for healthy NAROs and potential captive breeding. Observations and time spent at the Pueblo Zoo indicate that they have exhausted all means to achieve successful captive breeding between the males and the older female that they have housed since Their next option was to add a new female, and, late in 2012, the Pueblo Zoo did just that. The new female was just under a year old and originated in Louisiana. Copulation was noted between the new female and both male NAROs during 97

109 the 2013 breeding season. Although she is young, it is not unheard of for females to breed at such a young age. The new female did, in fact, become pregnant and gave birth to a litter of three pups in the spring of This information has been included in this study and coincides with the conclusion that having NAROs from similar geographic locations aids in successful captive breeding. 98

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118 Appendices A. Ethological Study of the Pueblo Zoo North American River Otters (Lontra canadensis) during the 2010 Breeding Season 97 B. Institutions Contacted for the Research Project 108 C. Surveys 124 D. Data 142 E. Pedigree Analysis 182 F. Internship Journals

119 Appendix A. Ethological Study of the North American River Otter (Lontra canadensis) during the 2010 Breeding Season Thelma - female Thor and Odin - males Sunday, 7 February 2010 NOTE: The female and males have been separated since November. This is done to mimic conditions in the wild where the sexes are only together during mating season Odin shows more interest in mating than Thor Thelma shows more interest in Thor 9:30 am - Arrive at zoo 10:00 am - Thelma and Odin are introduced to each other It starts snowing 10:02 am - They see each other Thelma presents herself to Odin when she sees him 10:05 am - They are doing a latrine dance Thelma is getting very vocal in response to his more aggressive efforts Thelma starts to bite at Odin and so he runs away 10:08 am - Odin approaches again; Thelma is making a lot of noise He grabs her neck and tries to mount her She is not receptive 10:09 am - Odin leaves again 10:12 am - Thelma presents herself 10:12.5 am - She moves away 10:13 am - Odin is examining where Thelma was a moment ago He smells all around to get her scent 10:15 am - Thelma is cleaning her ano-genital region and rubbing along the grass 10:20 am - They are both in the water and she does not run away 10:20.5 am - Odin gets out of the water and quickly goes back in 108

120 10:22 am - Odin pursues Thelma in the water, but she turns her head and runs out 10:25 am - Thelma is grooming in the dirt area of their enclosure The dirt helps remove oil and traps air in the fur. This helps keep the otters cleaner and warmer 10:26 am - Odin is smelling where Thelma presented herself He is very aggressively pursuing her in the water He follows her onto the land He gets a hold of her neck He is trying to mount and breed her 10:27 am - Thelma is screaming through all of Odin s advances, especially when he mounts her 10:28 am - They both do the latrine dance 10:37 am - Odin is smelling where Thelma was previously on her back and cleaning herself 10:38 am - Thelma is on her back again, outside of the dirt pile She is defecating for Odin to check out, but she is also rubbing herself all over the enclosure for him to smell 10:42 am - She is lying on her back again, showing some interest 10:44 am - Odin gets out of the water and makes his way toward Thelma She vocalizes and he stops 10:45 am - Odin moves a little closer Thelma vocalizes and he stops again 10:46 am - Odin goes to the places where Thelma was cleaning herself and lying on her back to get all of the smells NOTE: In past breeding seasons, Thelma usually tries to get as far away as possible from the boys. This time she is staying in one place, but she is vocalizing a lot to keep Odin away 10:50 am - Thelma ran toward Odin, vocalizing loudly He ran away 10:52 am - Thelma is lying on her back again, far away from Odin who is currently more interested in food 109

121 10:59 am - As Odin inches closer, Thelma starts vocalizing again He performs a latrine dance NOTE: Thelma is regularly presenting herself to both boys, but she is not receptive to either of them. 11:13 am - Thelma is in the water Odin is sniffing the areas she has been 11:14 am - Odin is pursuing Thelma in the water They are quickly out of the water and he has a hold of her neck She is vocalizing and he is grunting She is very unsettled, gets out of his neck hold, and quickly chases him away 11:37 am - Odin follows Thelma into the water She quickly gets out and vocalizes 11:38 am - Odin is not deterred by the vocalizations He pursues her, gets a good hold on her neck, and attempts to mate her She gets away quickly 11:39 am - Odin does the latrine dance He quickly rejoins her in the water 11:40 am - Thelma leaves the water 11:43 am - Odin follows Thelma out of the water He gets a hold of her neck and tries to mount her Thelma gets away and quickly chases Odin back into the water 12:00 pm - Stopped observations Tuesday, 9 February :00 am - Arrive at otter exhibit and begin observations 10:04 am - Thelma and Thor are on exhibit together Thor immediately begins mating rituals She allows him to smell her and she smells him 10:05 am - They go after the fish that has been placed around their enclosure instead of attempting any mating 10:24 am - Thelma and Thor are both in the water 110

122 Thor goes after her neck to attempt mating 10:25 am - They both get out of the water Thelma is presenting herself, but squirming around She is still on the ground, but he cannot get her neck She is checking out his privates and he is checking on her 10:27 am - Thor walks away and Thelma follows him 10:28 am - Thelma presents herself on land Thor is in the water 10:29 am - Thelma joins Thor in the water 10:30 am - There is prolonged togetherness in the water, some wrestling behavior 10:31 am - They go back on land and smell each other Almost immediately they go back in the water 10:33 am - Thor goes on land and Thelma follows him She bites his neck 10:37 am - They are both on land Both are presenting He is trying to get a hold of her throat 10:38 am - Thor is getting more intense and trying harder to get her neck 10:39 am - Thelma slips away Thor immediately pounces on Thelma as she tries to leave 10:40 am - Thor is thrusting, but not close enough to do anything She is grunting, not vocalizing as she did with Odin 10:41 am - Both otters leave each other and do their own thing 10:51 am - Both otters are in the water, but they are both more interested in watching me Thor leaves the water to find food Thelma is swimming circles by the window 10:56 am - Thor is in the water watching Thelma, but he is not pursuing her 11:00 am - Thor pursues Thelma in the water She just keeps swimming, so he leaves her alone 11:03 am - Thor is back in the water; once again, Thelma does not care 111

123 Thor is slowly following her in the water. When she moves to a new window, he follows, but she never seems to notice or care 11:00 am - I stepped away from directly in front of the exhibit 11:14 am - Thelma finally stops swimming circles in the windows She goes on land by Thor Both seem to be looking for food; they do not show any interest in each other 11:19 am - Thelma is rubbing herself by the fish-tree again The fish-tree is a smallish tree in the middle of the on-exhibit area that is perfect for the placement of fish at mealtime. It is easy for the otters to climb on this tree to reach different places. It is helpful for enrichment since it can hold different foods and requires some effort on the otters part NOTE: I have noticed that if I get too close to the window, Thelma will immediately go into the water to swim circles in front of the windows. 11:25 am - Thor is sleeping by the waterfall in the corner Thelma goes to Thor and smells his genitals Thor does not seem to care 11:27 am - Thelma nuzzles Thor and tries to get his attention, but he just sleeps 11:29 am - Thelma presents herself very close to him, but Thor does not move 11:34 am - Thelma is cleaning herself and rubbing against the rocks by Thor; again, all he does is sleep NOTE: Thelma is definitely more comfortable around Thor. They get along well and he is more passive than Odin. There has been no screaming on her part. Thor is more relaxed and does not seem as interested in mating with Thelma. Melanie, the otter keeper, says Thor is the more docile male. Odin tends to be more aggressive and the alpha male. If mating occurs and Thelma gets pregnant, Thor is unlikely to be the father. It is possible for Thelma to get pregnant by both males. Thelma is coming to the end of her breeding years. It is thought females stop breeding around age 10 and she is about 7.5 years old. 11:42 am - Both otters are sleeping in the waterfall area 11:47 am - Thor moved out of the sun, closer to Thelma She smelled him, then where he was lying 112

124 Now she is laying there 12:00 pm - Both still sleeping Thursday, 11 February :45 pm Arrive at zoo NOTE: Thor is on exhibit right now. He must know it is time to change enclosures, because he is waiting over by the door. Odin and Thelma are together again today. I will be observing them after they have been together several hours (instead of when they are first introduced). 12:57 pm - I can hear Thelma vocalizing and screaming. She is off-exhibit with Odin. I helped put out fish and crayfish on exhibit for their dinner. 1:20 pm - They are let into the exhibit. Both go after the cut up fish first, then the crayfish. They show no interest in each other 1:37 pm - Odin follows Thelma out of the water and pursues her He has a hold on her neck, but she is squirming She is very vocal and Odin lets go 1:38 pm - Both otters are in the water 1:43 pm - Odin began to follow Thelma, she swims away, so he gives up 1:45 pm - She goes back in the water and he goes to where she was on land and smells the area Odin does the latrine dance NOTE: Anytime Odin starts pursuing Thelma in the water, she quickly gets out 1:47 pm - Odin chases her out of the water and pursues her Thelma is vocalizing Odin is trying to grab her neck 1:48 pm - Thelma chases Odin away and she enters the water He goes to where they were wrestling and does the latrine dance 1:50 pm - Both otters are in the water, she is circling the window, and he is just relaxing 1:52 pm - He pursues her in the water and outside of the water 113

125 He is on top of her and has her neck She squirms out and chases him away 1:54 pm - Both get in the water again She is swimming in circles and he is watching 1:55 pm - Odin is looking for food again 1:56 pm - She vocalizes to him She is in the water, he is not, but after her vocalization he immediately jumps into the water 1:58 pm - He gets out again 2:00 pm - Odin lays down in the waterfall corner 2:05 pm - I have lost sight of Odin, but Thelma is still swimming circles in the window 2:10 pm - I have tried to move out of sight of Thelma, but she is still able to see me and therefore, she is still swimming in circles 2:20 pm - Odin is hiding and probably sleeping somewhere on exhibit I have not seen Odin since 2pm 2:20 pm - Thelma has stopped swimming in circles to enjoy a crayfish she found She is on land to eat Once done she is back in the water and swimming circles 2:21 pm - Thelma found another crayfish, so she is on land eating it 2:30 pm - I still have not seen Odin Thelma is still swimming in circles I need to come back next Tuesday to observe their second day together. Tuesday, 16 February 2010 Day 2 together for Thelma and Odin 9:40 am - Arrive at zoo NOTE: Melanie did not know that I was coming to observe the otters this morning until 9:30. She is disinfecting the patio and off-exhibit area. Thelma and Odin are in OH1, 2, and 3. Thelma is chirping and obviously upset. They are used to being out by now. Odin does not seem to be affected either way. 9:51 am - Thelma is on her back and Odin is smelling her 114

126 He is gently biting her neck She suddenly gets upset and fights him off Odin goes to OH 1; Thelma is in OH 3 Her chirping is louder and more frequent now 9:58 am - Thor is taken from on-exhibit and sent off-exhibit Odin, Thelma and Thor all touch noses through the gates before Thor continues to the off-exhibit area 10:03 am - Odin and Thelma are put on exhibit Both go directly into the water and then go after the food that has been put out Odin seems to be avoiding Thelma 10:15 am - Odin started diving for the PVC pipe of fish, but when Thelma went down for the same pipe, Odin immediately came back up and swam away 10:18 am - Thelma starts swimming in circles 10:31 am - Odin does the latrine dance and then goes in the water He is very docile in the water, often just floating Even when he is right next to Thelma, he just floats I try to walk away and still watch, but if Thelma sees me she starts to swim circles again 10:55 am - I could not get Thelma to stop swimming circles, so I left the exhibit to try to talk to Marilyn about goals for my internship. She was not in her office, so I returned to the exhibit 11:00 am - Upon returning to the exhibit I found Odin still sleeping in the same place Thelma was back by the waterfall and this time she did not get into the water when she saw me She was grooming herself and then laid down to sleep Thursday, 18 February :20 pm - Arrive at zoo NOTE: Today Thor and Thelma are together for the second day Fresh fish is put out for them Ashley, a zookeeper, is also putting out a lemon scent for enrichment 1:27 pm - Thelma and Thor are let out on exhibit 115

127 Both go straight for the food Thelma lies on her back in the water, floating, while eating fish 1:34 pm - Thelma has started swimming her circles 1:39 pm - No direct interaction between them, even when touching each other in the water 1:40 pm - Thor is interested in Thelma Thelma is swimming circles Thor seems to go after her neck 1:41 pm - Thelma swims her circles so Thor leaves the water 1:45 pm - So far, both otters have ignored or possibly not noticed the lemon scent 1:50 pm - I have stepped away from the exhibit and Thelma finally stops swimming in circles She goes onto the land. 1:52 pm - Thelma is back in the water when she sees me return Still no sign of Thor 1:54 pm - Some visitors came by the exhibit Thelma was very energetic and playful (she is quite the entertainer) 1:14 pm - Still no sign of Thor Thelma will not stop swimming in circles NOTE: There are considerably more visitors today. 2:27 pm - Even if I leave and Thelma gets out of the water, when she sees me return, she immediately gets back in and swims in circles Thursday, 25 February 2010 I missed the Tuesday visit because I was sick. 10:06 am - Arrive at zoo NOTE: Because of a misunderstanding, I am observing Thor and Thelma in their offexhibit area. 10:30 am - Thor and Thelma are both grunting; I am not sure if it is at me or each other Thor may have a cough They are both interested in the doors to the inside holding areas; Mel is cleaning 116

128 10:34 am - Thelma is very interested in me She follows me as I walk around the enclosure Thor is still sitting by the door to the inside 10:52 am - Thor and Thelma are rolling on each other and smelling each other 10:54 am - Thor is on his back sliding toward Thelma and smelling her 10:55 am - More grunting from Thor 10:57 am - Thelma and Thor are in the water She is in the water a lot, whereas Thor gets out sooner 11:02 am - Thor is scratching at the door Thelma is in the water bowl by the door She gets out and starts to smell Thor She is cleaning herself right next to Thor He is oblivious that Thelma is right next to him 11:24 am - Thor comes into the exhibit, away from the door He goes to check out Thelma, then walks toward the water Thelma is too interested in me to notice Thor 11:25 am - Thor comes closer to the bottom of the exhibit 11:26 am - Thor smells Thelma She walks away, then follows him after he leaves 11:27 am - Thor is back at the door that leads inside NOTE: I left the otters in back at 11:30 am. I stopped by the on-exhibit to see Odin. When he saw me, he came into the water and swam a little. He looked at me and stayed where I could see him. He is not an energetic entertainer like Thelma. Friday, 26 February 2010 NOTE: I stopped by the otter exhibit when I visited the zoo with family. Thelma was in the water showing off. Odin was with Thelma. He was very curious to see me and my family. He spent time in the water watching and following me. He didn t try anything with Thelma. Thursday, 4 March :30 am - Arrive at the zoo 117

129 NOTE: The otters are already on exhibit. 10:31 am - Thelma sees me and immediately jumps into the water She is swimming circles, but slower than normal Odin is sleeping in one of the underground nests; I can see him curled up 10:40 am - A guest came by and Thelma was more energetic Odin has not moved 10:45 am - I moved to the left side of the exhibit to a bench in the sun Thelma got very excited; she swam faster circles and jumped 11:03 am - Odin left the den 11:06 am - Odin does a latrine dance 11:07 am - Odin is in the water and checking out zoo guests Both otters are in the water 11:20 am - Odin on land or in the water continues to watch the guests, NOTE: Mel told me that Odin was slightly afraid to be in an enclosure with Thelma. His best efforts at mating have been shot down and he had been beaten up by her. Thelma and Thor are best buddies. 11:27 am - Thor is spending a lot of time in the water; more than I usually observe. NOTE: Even though Odin and Thelma are in the water together, Odin is staying away from her. 11:29 am - Odin is out of the water and sunning himself 11:32 am - More zoo guests arrive Odin jumps into the water 11:33 am - Odin gets out 11:42 am - Thelma finally leaves the water Just as Thelma gets out, Megan, a zookeeper, walks by and both otters get back in the water and follow her as she walks by 11:46 am - Odin gets out of the water Thelma is swimming circles again 11:55 am - More zoo guests arrive 118

130 Both otters get in the water and start entertaining the guests 12:00 pm - I stop observations and go talk to Marilyn 119

131 Appendix B. Institutions Contacted for the Research Project I. from David Hamilton II. III. IV. Reminder from Researcher Phone Message from Researcher Tables of Institution Contacts B1. Association of Zoos and Aquariums accredited successful breeding institutions B2. Association of Zoos and Aquariums accredited unsuccessful captive breeding institutions B3. Association of Zoos and Aquariums accredited non-breeding institutions B4. Association of Zoos and Aquariums accredited institutions who did not respond B5. Non-Association of Zoos and Aquariums accredited institutions 120

132 I. from David Hamilton The completed survey was ed by David Hamilton, studbook coordinator, to the Association of Zoos and Aquariums list of establishments that house North American River Otters (NAROIR). The text of his is shown below. North American River Otter IRs, Please take the time to fill out the survey in the SurveyMonkey link below. It is a project by Allison Haar. She is a graduate student at Colorado State University at Pueblo working towards a Masters in Biology. She is studying the captive breeding of the North American river otters. Hopefully she will be able to tease out what those successful institutions are doing so that the rest of us can replicate those practices. All participating zoos will receive a copy of the results from Allison. Thanks for your support of this important project. I also have another student looking at the latitude question. I hope to be sharing those results at the workshop, Sincerely, David David B. Hamilton General Curator Seneca Park Zoo 2222 St. Paul Street Rochester, NY Office: ***.***.**** Mobile: ***.***.**** Fax:: ***.***.**** dhamilton@monroecounty.gov 121

133 II. Reminder from researcher The text of the reminder sent by the researcher to the NAROIR list is shown below. Dear NAROIRs, My name is Allison Haar and I am a masters student at Colorado State University- Pueblo. Approximately two months ago you received an from David Hamilton encouraging everyone to complete a survey about North American River Otter captive breeding on survey monkey for my masters research project. I thank everyone who previously completed the survey and I am writing now to ask everyone else to please respond, as I do not have enough responses to complete the statistical research. Please take a few minutes and complete the survey as thoroughly as possible, especially those zoos that have been unsuccessful with their NARO breeding programs. I need these answers as much as those from successful zoos. Please follow the link to complete the survey: Thank you for your time and cooperation, Allison Haar ak.haar@colostate-pueblo.edu ***.***.**** 122

134 III. Phone message from researcher The text of the phone message left for respondents when a zoo or aquarium was contacted is found below. Hello my name is Allison Haar. I am a graduate student at Colorado State University in Pueblo, Colorado. I am doing a masters research project on factors that contribute to successful captive breeding of the North American River Otter. If you have tried to breed your otters I would appreciate if you could complete a short survey. Please call me at ***.***.**** or me at ak.haar@colostate-pueblo.edu, at your earliest convenience. Again my name is Allison asking about your river otters. Thank you for your time. 123

135 IV. Contacted Institutions B1. Association of Zoos and Aquariums accredited successful breeding institutions B2. Association of Zoos and Aquariums accredited unsuccessful captive breeding institutions B3. Association of Zoos and Aquariums accredited non-breeding institutions B4. Association of Zoos and Aquariums accredited institutions who did not respond B5. Non- Association of Zoos and Aquariums accredited institutions 124

136 Table B1. List of Association of Zoos and Aquariums accredited successful captive breeding institutions who completed the study survey, showing when, how, and by whom the institutions were contacted. from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher Name of institution Location Alexandria Zoological Park Alexandria, LA 6/9/11 6/22/11 7/6/11 8/11/11 Binghamton Zoo at Ross Park Binghamton, NY 6/9/11 6/22/11 7/6/11 8/11/11 Brookgreen Gardens Zoo Pawleys Island, SC 6/9/11 6/22/11 7/6/11 8/11/11 Cincinnati Zoo and Botanical Cincinnati, OH Personal from researcher Last contact Phone call from researcher Gardens 6/9/11 6/22/11 7/6/11 8/11/11 10/4/11 Columbus Zoo and Aquarium Powell, OH 6/9/11 6/22/11 7/6/11 8/11/11 Denver Zoological Gardens Denver, CO 6/9/11 6/22/11 7/6/11 8/11/11 Dickerson Park Zoo Springfield, MO 6/9/11 6/22/11 7/6/11 8/11/11 11/18/12 11/13/12 Lee Richardson Zoo Garden City, KS 6/9/11 6/22/11 7/6/11 8/11/11 10/7/12 Little Rock Zoological Gardens Little Rock, AR 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 11/13/12 Minnesota Zoological Garden Apple Valley, MN 6/9/11 6/22/11 7/6/11 8/11/11 Oakland Zoo Oakland, CA 6/9/11 6/22/11 7/6/11 8/11/11 Palm Beach Zoo at Dreher Park West Palm Beach, FL 6/9/11 6/22/11 7/6/11 8/11/11 Pueblo Zoo Pueblo, CO 6/9/11 6/22/11 7/6/11 8/11/11 Sedgwick County Zoo Wichita, KS 6/9/11 6/22/11 7/6/11 8/11/11 11/28/11 10/25/12 10/25/12 125

137 Table B2. List of Association of Zoos and Aquariums accredited unsuccessful captive breeding institutions who completed the study survey, showing when, how, and by whom the institutions were contacted. from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher Personal from researcher Other Name of institution Location contact Buffalo Zoological Gardens Buffalo, NY 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 10/7/12 Connecticut's Beardsley Zoo Bridgeport, CT 6/9/11 6/22/11 7/6/11 8/11/11 11/28/11 10/25/12 Dakota Zoo Bismarck, ND 6/9/11 6/22/11 7/6/11 8/11/11 Folsom Children's Zoo Lincoln, NE 6/9/11 6/22/11 7/6/11 8/11/11 Fort Worth Zoological Park Fort Worth, TX 6/9/11 6/22/11 7/6/11 8/11/11 Hutchinson Zoo Hutchinson, KS 6/9/11 6/22/11 7/6/11 8/11/11 Kansas City Zoo Kansas City, MO 6/9/11 6/22/11 7/6/11 8/11/11 Lincoln Park Zoological Gardens Chicago, IL 6/9/11 6/22/11 7/6/11 8/11/11 North Carolina Zoological Park Asheboro, NC 6/9/11 6/22/11 7/6/11 8/11/11 Oregon Zoo Portland, OR 6/9/11 6/22/11 7/6/11 8/11/11 Pueblo Zoo Pueblo, CO 6/9/11 6/22/11 7/6/11 8/11/11 Roosevelt Park Zoo Minot, ND 6/9/11 6/22/11 7/6/11 8/11/11 Rosamond Gifford Zoo at Burnet Park Syracuse, NY 6/9/11 6/22/11 7/6/11 8/11/11 Seneca Park Zoo Rochester, NY 6/9/11 6/22/11 7/6/11 8/11/11 Texas State Aquarium Corpus Christi, TX 6/9/11 6/22/11 7/6/11 8/11/11 Trevor Zoo Millbrook, NY 6/9/11 6/22/11 7/6/11 8/11/11 Phone call from researcher 126

138 Table B3. List of Association of Zoos and Aquariums accredited institutions that do not breed their North American river otters, but completed the study survey, showing when, how, and by whom the institutions were contacted. from David Hamilton 127 with fixed survey link Reminder from David Hamilton Reminder from researcher Personal from researcher Phone call from researcher Last Name of institution Location contact Audubon Zoo New Orleans, LA 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Birmingham Zoo Birmingham, AL 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 Brandywine Zoo Wilmington, DE 6/9/11 6/22/11 7/6/11 8/11/11 11/15/12 11/13/12 Brec's Baton Rouge Zoo Baker, LA 6/9/11 6/22/11 7/6/11 8/11/11 11/15/12 11/13/12 Caldwell Zoo Tyler, TX 6/9/11 6/22/11 7/6/11 8/11/11 10/4/12 Capron Park Zoo Attleboro, MA 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Chahinkapa Zoo Wahpeton, ND 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 10/7/12 Downtown Aquarium Denver, CO 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Ecotarium Worchester, MA 6/9/11 6/22/11 7/6/11 8/11/11 11/20/12 11/19/12 John G. Shedd Aquarium Chicago, IL 6/9/11 6/22/11 7/6/11 8/11/11 10/4/12 Knoxville Zoological Knoxville, TN Gardens 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Los Angeles Zoo and Los Angeles, CA Botanical Gardens 6/9/11 6/22/11 7/6/11 8/11/11 Milwaukee County Zoo Milwaukee, WI 6/9/11 6/22/11 7/6/11 8/11/11 North Carolina Aquarium at Pine Knoll Shores Northwest Trek Wildlife Park Omaha's Henry Doorly Zoo and Aquarium Oregon High Desert Museum Atlantic Beach, NC 6/9/11 6/22/11 7/6/11 8/11/11 Eatonville, WA Omaha, NE Bend, OR 6/9/11 6/22/11 7/6/11 8/11/11 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12

139 Table B3 cont d. List of Association of Zoos and Aquariums accredited institutions that do not breed their North American river otters, but completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution Location from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher Personal from researcher San Francisco Zoological San Francisco Gardens 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Seattle Aquarium Seattle, WA 6/9/11 6/22/11 7/6/11 8/11/11 11/04/12 Woodland Park Zoological Seattle, WA Gardens 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 Last contact Phone call from researcher 128

140 Table B4. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution Location from David Hamilton link Hamilton Reminder from researcher Akron Zoological Park Akron, OH 6/9/11 6/22/11 7/6/11 8/11/11 Alameda Park Zoo 129 with fixed survey Reminder from David Personal from researcher Last contact Phone call from researcher Alamogordo, NM 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 Alaska Zoo Anchorage, AK 6/9/11 6/22/11 7/6/11 8/11/11 Arizona-Sonora Desert Tucson, AZ Museum 6/9/11 6/22/11 7/6/11 8/11/11 10/2/12 Bays Mountain Park Kingsport, TN 6/9/11 6/22/11 7/6/11 8/11/11 Montreal, Biodome de Montreal Quebec 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Blank Park Zoo of Des Des Moines, IA Moines 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Boonshoft Museum of Dayton, OH Discovery 6/9/11 6/22/11 7/6/11 8/11/11 Brevard Zoo Melbourne, FL 6/9/11 6/22/11 7/6/11 8/11/11 Buttonwood Park Zoo New Bedford, MA 6/9/11 6/22/11 7/6/11 8/11/11 11/28/11 Calgary Zoo Calgary, Alberta 6/9/11 6/22/11 7/6/11 8/11/11 10/2/12 Calvert Marine Museum Solomons, MD 6/9/11 6/22/11 7/6/11 8/11/11 Cameron Park Zoo Waco, TX 6/9/11 6/22/11 7/6/11 8/11/11 Cape May County Park Zoo Cape May Court House, NJ 6/9/11 6/22/11 7/6/11 8/11/11 Central Park Zoo New York, NY 6/9/11 6/22/11 7/6/11 8/11/11

141 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution Cheyenne Mountain Zoological Park Location from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher Personal from researcher Last contact Phone call from researcher Colorado Springs, CO 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Chicago Zoological Park Brookfield, IL (Brookgreen Gardens) 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Clearwater Marine Science Clearwater, FL Center 6/9/11 6/22/11 7/6/11 8/11/11 Clinch Park Zoo and Aquarium Traverse City, MI 6/9/11 6/22/11 7/6/11 8/11/11 Coyote Point Museum San Mateo, CA 6/9/11 6/22/11 7/6/11 8/11/11 Detroit Zoological Institute Royal Oak, MI 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 11/19/12 Ellen Trout Zoo Lufkin, TX 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 11/19/12 Elmwood Park Zoo Norristown, PA 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Erie Zoological Gardens Erie, PA 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Florida Aquarium Tampa, FL 6/9/11 6/22/11 7/6/11 8/11/11 Fort Rickey Children's Discovery Zoo Rome, NY 6/9/11 6/22/11 7/6/11 8/11/11 Fort Wayne Children's Fort Wayne, IN Zoological Garden 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Henry Villas Zoo Madison, WI 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Henson Robinson Zoo Springfield, IL 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Houston Zoo Houston, TX 6/9/11 6/22/11 7/6/11 8/11/11 130

142 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. from David Hamilton 131 with fixed survey link Reminder from David Hamilton Reminder from researcher Name of institution Location Interzoo Laval, Quebec 6/9/11 6/22/11 7/6/11 8/11/11 Jackson Zoological Park Jackson, MS 6/9/11 6/22/11 7/6/11 8/11/11 Jacksonville Zoo and Gardens Jacksonville, FL 6/9/11 6/22/11 7/6/11 8/11/11 John Ball Zoological Garden Personal from researcher Last contact Grand Rapids, MI 6/9/11 6/22/11 7/6/11 8/11/11 11/28/11 10/3/12 Lake Superior Zoological Gardens Duluth, MN 6/9/11 6/22/11 7/6/11 8/11/11 Lehigh Valley Zoo Phone call from researcher Schnecksville, PA 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Maritime Center at Norwalk South Norwalk, CT 6/9/11 6/22/11 7/6/11 8/11/11 Mesker Park Zoo Evansville, IN 6/9/11 6/22/11 7/6/11 8/11/11 Miller Park Zoo Bloomington, IL 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Montgomery Zoo Montgomery, AL 6/9/11 6/22/11 7/6/11 8/11/11 New York State Zoo at Watertown, NY Thompson Park 6/9/11 6/22/11 7/6/11 8/11/11 NEW Zoo Green Bay, WI 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Niabi Zoo Coal Valley, IL 6/9/11 6/22/11 7/6/11 8/11/11 North Carolina Aquarium on Roanoke Island Manteo, NC 6/9/11 6/22/11 7/6/11 8/11/11

143 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution Oglebay's Good Children's Zoo Location Wheeling, WV from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher 6/9/11 6/22/11 7/6/11 8/11/11 Personal from researcher Last contact Phone call from researcher Oklahoma City Zoological Park Oklahoma City, OK 6/9/11 6/22/11 7/6/11 8/11/11 Pittsburgh Zoo and PPG Pittsburgh, PA Aquarium 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Potter Park Zoological Lansing, MI Gardens 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Prospect Park Zoo Brooklyn, NY 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Provincial Wildlife Park Shubenacadie, Nova Scotia 6/9/11 6/22/11 7/6/11 8/11/11 Racine Zoological Gardens Racine, WI 6/9/11 6/22/11 7/6/11 8/11/11 Riverside Zoo Scottsbluff, NE 6/9/11 6/22/11 7/6/11 8/11/11 Rocky Mountain Ark Wildlife Rehabilitation Telluride, CO Center 6/9/11 6/22/11 7/6/11 8/11/11 Sacramento Zoo Sacramento, CA 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Saint Louis Zoological Park St. Louis, MO 6/9/11 6/22/11 7/6/11 8/11/11 11/28/11 Salisbury Zoological Park Salisbury, MD 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 132

144 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution San Antonio Zoological Gardens and Aquarium Location from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher San Antonio, TX 6/9/11 6/22/11 7/6/11 8/11/11 Sciworks (Science center and Environmental Park) Winston-Salem, NC 6/9/11 6/22/11 7/6/11 8/11/11 Silver Springs Park Silver Springs, FL 6/9/11 6/22/11 7/6/11 8/11/11 Smithsonian National Zoological Park Washington, DC 6/9/11 6/22/11 7/6/11 8/11/11 South Carolina Aquarium Charleston, SC 6/9/11 6/22/11 7/6/11 8/11/11 Stamford Museum and Nature Center Stamford, CT 6/9/11 6/22/11 7/6/11 8/11/11 Personal from researcher Last contact Phone call from researcher Tallahassee Museum of Tallahassee, FL History and Natural Sciences 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Tampa's Lowry Park Zoo Tampa, FL 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Tautphaus Park Zoo Idaho Falls, ID 6/9/11 6/22/11 7/6/11 8/11/11 11/13/12 Tennessee Aquarium Chatanooga, TN 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Texas Zoo Victoria, TX 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 The Maryland Zoo in Baltimore, MD Baltimore 6/9/11 6/22/11 7/6/11 8/11/11 The Zoo, Northwest Florida Gulf Breeze, FL 6/9/11 6/22/11 7/6/11 8/11/11 133

145 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. with fixed survey Reminder from David Name of institution Location from David Hamilton link Hamilton Reminder from researcher Topeka Zoo Topeka, KS 6/9/11 6/22/11 7/6/11 8/11/11 Personal from researcher Last contact Phone call from researcher Toronto Zoo Scarborough, Ontario 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 Tulsa Zoo and Living Museum Tulsa, OK 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 West Orange, Turtle Back Zoo NJ 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 Virginia Aquarium and Marine Science Center Virginia Living Museum Virginia Beach, VA 6/9/11 6/22/11 7/6/11 8/11/11 Newport News, VA 6/9/11 6/22/11 7/6/11 8/11/11 Western North Carolina Asheville, NC Nature Center 6/9/11 6/22/11 7/6/11 8/11/11 Wildlife Prairie State Park Peoria, IL 6/9/11 6/22/11 7/6/11 8/11/11 Springfield, Wonders of Wildlife Museum MO 6/9/11 6/22/11 7/6/11 8/11/11 Zoo Boise Boise, ID 6/9/11 6/22/11 7/6/11 8/11/11 Zoo New England/Franklin Boston, MA Park Zoo 6/9/11 6/22/11 7/6/11 8/11/11 10/8/12 Zoo Sauvage de St.-Félicien St. Félicien, Quebec 6/9/11 6/22/11 7/6/11 8/11/11 10/3/12 ZooAmerica Hershey, PA 6/9/11 6/22/11 7/6/11 8/11/11 11/19/12 134

146 Table B4 cont d. List of Association of Zoos and Aquariums accredited institutions that were contacted, but never completed the study survey, showing when, how, and by whom the institutions were contacted. Name of institution Location from David Hamilton with fixed survey link Reminder from David Hamilton Reminder from researcher Personal from researcher Last contact Phone call from researcher Zoological Society of San San Diego, CA Diego 6/9/11 6/22/11 7/6/11 8/11/11 ZooMontana Billings, MT 6/9/11 6/22/11 7/6/11 8/11/11 10/25/12 Table B5. List of non - Association of Zoos and Aquariums accredited institutions that were contacted to complete the study survey, showing when, how, and by whom the institutions were contacted. Personal from researcher Phone call from researcher Successful breeders / Unsuccessful breeders Survey Name of institution Location Complete Flamingo Gardens Davie, FL 11/29/12 Yes Successful Edmonton Valley Zoo Edmonton, Alberta 10/5/12 10/2/12 No No response Homosassa Springs Wildlife State Park Homosassa, FL 11/13/12 No No response Staten Island Zoo New York, NY 11/19/12 No No response The Florida Aquarium Tampa, FL 11/13/12 No No response Zoo Montana Billings, MT 11/25/12 No No response Zoo of Acadiana Broussard, LA 11/13/12 No No response 135

147 Appendix C. Surveys I. North American River Otter Questionnaire 1 II. North American River Otter Questionnaire 2 III. North American River Otter Questionnaire 3 136

148 I. North American River Otter Questionnaire 1 My name is Allison Haar. I am a graduate student at Colorado State University at Pueblo working towards a Masters in Biology. For my research project, I am conducting a study on the captive breeding of the North American River Otter (Lontra canadensis). Using this questionnaire, I will compare and statistically analyze the individual otters as well as zoological practices. I hope to gain useful and applicable knowledge to improve captive breeding techniques. This questionnaire has been approved by David Hamilton and is sanctioned by the SSP. All participating zoos will receive a copy of the results. QUESTION 1 1. Please state your name, institution, job title and contact information. Otter Details 2. Female(s) age, average weight and length if possible 3. Male age(s), average weight and length if possible QUESTION 2 Geography 1a. What is the ancestry of your otter(s)? 1b. Is the otter wild-caught or captive born? 1c. If wild-caught, what is the area of origination (state, region or latitude if known)? 2. How long have your otters been at your zoo? 1a. 1b. 1c

149 QUESTION 3 Successful Breeding 1a. How many litters has the female had? 1b. What is the known age of the female when she had her first litter? 1c. Are the litters with the same or different males? 1d. Number of surviving young per litter. 2. How many years were the male and female together before they successfully mated? 3. How many pseudo-pregnancies have occurred? 4. Have any of your otters been contracepted? If so, when? 1a. 1b. 1c. 1d QUESTION 4 Housing 1. How many males and females are together on exhibit? 2a. Are the males and females housed together or separately? 2b. Is the housing arrangement year round or seasonal? If seasonal, please explain. 2c. If separated do they have access to each other s areas or in completely separate locations?

150 2a. 2b. 2c. QUESTION 5 Diet and Enrichment 1. Please describe the daily otter diet. 2. Please describe enrichment foods. 3. Please list all vitamins and supplements QUESTION 6 Exhibit What are the specifics of the otter exhibit? 1a. Square footage: 1b. % water: land area 1c. Types of vegetation: 2. Is there an off-exhibit and if so what are the specifics of the off exhibit? 3. Do they have access outside at night? 4. Do they have access to a large pool of water both on and off exhibit? 5. How close is the pool to public viewing? 1a. 139

151 1b. 1c QUESTION 7 Den/Nestbox Please describe the type of den/nestbox provided for the female. 1a. Dimensions: 1b. Types of material: 1c. Indoors/outdoors: 1d. Diggable or a pen area: 2. Is more than one den/nestbox available? 3. When does the female have access to the den/nestbox area? For example, all year or seasonally during pupping season. 1a. 1b. 1c. 1d

152 QUESTION 8 4. What kind of nesting material is provided? 5. How does the type and amount of nesting material differ between the breeding and non-breeding seasons? 6. Does the female have her own outdoor area, where only her scent would be found? QUESTION 9 Keeper Involvement 1a. How much social interaction do the keepers have with the otters? 1b. Is there direct contact? 2. Do you think your keeper/otter relationship has an impact on reproduction? 1a. 1b

153 II. North American River Otter Questionnaire 2 My name is Allison Haar. I am a graduate student at Colorado State University at Pueblo working towards a Masters in Biology. For my research project, I am conducting a study on the captive breeding of the North American River Otter (Lontra canadensis). Using this questionnaire, I will compare and statistically analyze the individual otters as well as zoological practices. I hope to gain useful and applicable knowledge to improve captive breeding techniques. This questionnaire has been approved by David Hamilton and is sanctioned by the SSP. All participating zoos will receive a copy of the results. QUESTION 1 Please state your name, institution and contact information. Name: Institution: Address: Address 2: City/Town: State: ZIP: Country: Address: Phone Number: QUESTION 2 Please enter your job title QUESTION 3 Please enter the following information related to your female otter(s). Age. Average Weight. Average Length. QUESTION 4 142

154 Please enter the following information related to your male otter(s). Age. Average weight. Average length. QUESTION 5 Geography With regard to your female otter(s: What is the ancestry of your female otter(s)? Is/Are the otter(s) wild-caught or captive born? If wild-caught, what is the area of origination (state, region or latitude if known)? QUESTION 6 Geography How long has/have your female otter(s) been at your facility? Less than 1 year. > 1 to < 3 years. > 3 to < 5 years. More than 5 years. QUESTION 7 Geography In regard to your male otter(s): What is the ancestry of your male otter(s)? Is/Are the otter(s) wild-caught or captive born? If wild-caught, what is the area of origination (state, region or latitude if known)? QUESTION 8 143

155 How long has/have your male otter(s) been at your facility? Less than 1 year. > 1 to < 3 years. > 3 to 5 years. More than 5 years. QUESTION 9 Successful Breeding How many litters has the female had? None. One. Two. Three. More than 3. QUESTION 10 Successful Breeding What is the known age of the female when she had her first litter? QUESTION 11 Successful Breeding Are the litters with the same or different male(s)? Same. Different. QUESTION 12 Successful Breeding Number of young per litter more than 6 144

156 Litter 1 Litter 2 Litter 3 Other litters QUESTION 13 Successful Breeding Number of surviving young per litter. Litter 1 Litter 2 Litter 3 Other litters more than 6 QUESTION 14 Successful Breeding How many years were the male and female together before they successfully mated? More than Years. QUESTION 15 Successful Breeding How many pseudo-pregnancies have occurred? More than 3 Pseudopregnancy QUESTION 16 Successful Breeding 145

157 Have any of your otters been contracepted? If so, when? Yes. No. When did this occur? QUESTION 17 Housing How many males and females are together on exhibit? More than 4 Female. Male. QUESTION 18 Housing Are the males and females housed together or separately? Together. Separately. QUESTION 19 Housing Is the housing arrangement year round or seasonal? If seasonal, please explain. Year round. Seasonal. Comment if seasonal arrangement QUESTION 20 Housing If separated do they have access to each other s areas or are these completely separate locations? Access to each other's areas. Completely separate locations. 146

158 QUESTION 21 Diet and Enrichment Please describe the daily otter diet. QUESTION 22 Diet and Enrichment Please describe enrichment foods. QUESTION 23 Diet and Enrichment Please list all vitamins and supplements. QUESTION 24 Exhibit What are the specifics of the otter exhibit? Square Footage. % water/land area. Types of vegetations. QUESTION 25 Exhibit Is there an off-exhibit? Yes. No. Describe the off-exhibit. QUESTION 26 Exhibit 147

159 Do the otters have access outside at night? Yes. No. QUESTION 27 Exhibit Do the otters have access to a large pool of water both on and off exhibit? Yes, on-exhibit. No, on-exhibit. Yes, off-exhibit. No, off-exhibit. QUESTION 28 Exhibit How close is the pool to public viewing? Less than 5 feet. 5 feet to 15 feet. More than 15 feet. QUESTION 29 Den/Nestbox Please describe the type of den/nestbox provided for the female. Dimensions. Types of material. Indoors/Outdoors. Diggable or a pen area. Number of available dens/nestboxes. QUESTION 30 Den/Nestbox 148

160 When does the female have access to the den/nestbox area? For example, all year or seasonally during pupping season. All year. Seasonally. Other. If other, please describe the timing. QUESTION 31 Den/Nestbox What kind of nesting materials are provided? QUESTION 32 Den/Nestbox Does the type and amount of nesting material differ between the breeding and nonbreeding seasons? Yes, nesting material differs between seasons. No, nesting material does not differ between seasons. If yes, in what way do they differ? QUESTION 33 Den/Nestbox Does the female have her own outdoor area, where only her scent would be found? Yes. No. QUESTION 34 Keeper Involvement How much social interaction do the keepers have with the otters? Minimal, < 1/2 hour daily. Limited, > 1/2 up to 1 hour daily. 149

161 Extensive, > 1 hour daily. Rationale for degree of interaction QUESTION 35 Keeper Involvement Is there direct contact between the Keeper and the otter(s)? Yes. No. Type of contact and rationale for contact. QUESTION 36 Keeper Involvement Do you think your keeper/otter relationship has an impact on reproduction? Yes. No. Why? 150

162 III. North American River Otter Questionnaire 3 My name is Allison Haar. I am a graduate student at Colorado State University at Pueblo working towards a Masters in Biology. For my research project, I am conducting a study on the captive breeding of the North American River Otter (Lontra canadensis). Using this questionnaire, I will compare and statistically analyze the individual otters as well as zoological practices. I hope to gain useful and applicable knowledge to improve captive breeding techniques. This questionnaire has been approved by David Hamilton and is sanctioned by the SSP. All participating zoos will receive a copy of the results. QUESTION 1 Name Institution Job Title Phone Number Address QUESTION 2 Female Otter(s) Breeding female age. Breeding female weight. Is the otter wild-caught or captive born? What is the area of origination (zoo, state, region or latitude, if known)? QUESTION 3 Male Otter(s) Breeding male age. Breeding male weight. Is the otter wild-caught or captive born? What is the area of origination (zoo, state, region or latitude if known)? 151

163 QUESTION 4 Successful Breeding Information What is the known or approximate age of the female when she had her first litter? Please state the number of young born in each litter (ex. 1-3, 2-2, 3-5). How many years were the male and female together before they successfully mated? QUESTION 5 Diet and Enrichment Please describe the daily diet. Please describe enrichment foods. Please list all vitamins and supplements. QUESTION 6 Housing How many males and females are together on exhibit? Are the males and females housed together or separately? If separated, do they have access to each others' areas? Is the housing arrangement year round or seasonal? If seasonal, please explain. QUESTION 7 Exhibit Is there an off-exhibit? Do they have access outside at night? Do they have access to a large pool of water both on and off exhibit? 152

164 QUESTION 8 Den/Nestbox Please describe the den/nestbox material: Is the den/nestbox indoors or outdoors: Is it a diggable area or a pen area? Number of dens/nestboxes available? Is access to dens/nestboxes given all year or seasonally? Please describe the nesting materials provided? QUESTION 9 Keeper Involvement Approximately how much interaction occurs between the otters and the keepers on a daily basis? Is there direct contact between keepers and otters? Do you think your keeper/otter relationship has an impact on reproduction? 153

165 Appendix D. Data Table D1. Successful Captive Breeding Institution Data Table D2. Unsuccessful Captive Breeding Institution Data 154

166 Table D1. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Alexandria Zoological Park 4 Louisiana Wild 16 Binghamton Zoo at Ross Park 14 Unknown Captive 4.33 Brookgreen Gardens 5 Florida Wild 5 Cincinnati Zoo & Botanical Garden 14 Nashville, TN Captive 14 Columbus Zoo & Aquarium 18 St. Louis Zoo, St. Louis, MO Captive 10 Denver Zoo 13 Louisiana Wild

167 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of Female Captive born or Wild caught Age of female when she had her first litter Number of litters Alexandria Zoological Park Louisiana Wild 3 1 Binghamton Zoo at Ross Park Beardsley Zoo, Bridgeport, Ct Captive 3 1 Brookgreen Gardens South Carolina Captive Cincinnati Zoo & Botanical Garden Nashville, TN Captive 8 1 Columbus Zoo & Aquarium Paducah, KY Wild 5 3 Denver Zoo Florida Wild

168 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Number of young per litter Number of years the male and female were together before successful captive breeding Male and female together Year-round or Seasonally Foods included in the regular daily diet Alexandria Zoological Park 3 1 Year-round capelin fish, herring fish, AAA brand ground meat Binghamton Zoo at Ross Park 2 3 Year-round fish, meateater, produce Brookgreen Gardens 3 Cincinnati Zoo & Botanical Garden Unknown, eaten at birth 8 Year-round capelin, herring, Nebraska feline diet, Purina cat chow, hard boiled, egg, mouse Columbus Zoo & Aquarium 3, 3, 3 2 Year-round Nebraska carnivore diet, lake smelt, capelin, trout Denver Zoo 2 Year-round fish, Toronto brand small carnivore diet 157

169 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Enrichment foods Daily supplements Is the den indoors or outdoors and is it on exhibit or off exhibit? Is the den a pen or diggable area? Alexandria Zoological Park Binghamton Zoo at Ross Park crickets, squid, feeder fish, bloodsicles, rodents, produce Pet Tab vitamin E and vitamin B1 Indoors Pen 50mg vitamin B1, 1000 IU vitamin E Indoors Pen Brookgreen Gardens Cincinnati Zoo & Botanical Garden live fish, clams, shrimp thiamin E paste Indoors Pen Columbus Zoo & Aquarium Denver Zoo clams, krill, live fish, frozen fish, scallops, shrimp, mice ½ g thiamin E Paste 2 Indoors, 2 Off exhibit All are pen areas tuna smears, peanut butter smears, cat food smears, melons, or other fruit or veggie pieces vitamin E, vitamin B1 Indoors, Off exhibit no 158

170 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Number of dens Nest materials provided Are the nesting materials seasonal? If seasonal, how do the nesting materials differ? Alexandria Zoological Park 3 hay, grass they bring indoors Yes They have materials in winter, not in summer Binghamton Zoo at Ross Park 3 hay Yes hay Brookgreen Gardens Cincinnati Zoo & Botanical Garden 2 wood shavings, straw Columbus Zoo & Aquarium 4 straw Yes straw Denver Zoo 1 grass hay No 159

171 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Alexandria Zoological Park Medium (> 1/2 up to 1 hr) No Binghamton Zoo at Ross Park Medium (> 1/2 up to 1 hr) No Brookgreen Gardens Cincinnati Zoo & Botanical Garden Medium (> 1/2 up to 1 hr) [30-40 min] No Unknown Columbus Zoo & Aquarium High (> 1 hr) No Yes Denver Zoo Medium (> 1/2 up to 1 hr) Yes No 160

172 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Dickerson Park Zoo 3 Tulsa, OK Wild 7 Flamingo Gardens 10 St. Augustine, FL Wild 28 Lee Richardson Zoo 4 Baton Rouge, LA Wild 9 Little Rock Zoo 15 Nisswa, MN Wild 12 Minnesota Zoo 10 Florida Captive 8 Oakland Zoo 12 Florida Wild 4 161

173 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of Female Captive born or Wild caught Age of female when she had her first litter Number of litters Dickerson Park Zoo Taney County, MO Wild 4 5 Flamingo Gardens Florida Wild 4 Lee Richardson Zoo Dickerson Park Zoo, Springfield, MO Captive 8 1 Little Rock Zoo Montreal, Canada Wild 6 3 Minnesota Zoo Sedgwick County Zoo, Wichita, KS Captive 7 1 Oakland Zoo Buttonwood Park Zoo, New Bedford, MA Captive

174 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Number of young per litter Number of years the male and female were together before successful captive breeding Male and female together Year-round or Seasonally Foods included in the regular daily diet Dickerson Park Zoo 3, 4, 4, 4, 4 4, 2 Year-round Flamingo Gardens less than 1 Year-round Lee Richardson Zoo 4 1 Little Rock Zoo 3, 4, 4 Seasonal Minnesota Zoo 4 2 Seasonal Oakland Zoo 2 2 Year-round meat, fish (mackerel, herring, smelt) capelin, smelt, herring, raw meat Nebraska special beef, carrot, smelt Nebraska feline diet, dry polar bear diet Toronto carnivore diet, fish capelin, Natural Balance 5, Mazuri Ferret Diet, hard-boiled eggs, chicks, bones, mealworms/crickets, goldfish, mice, fruit/vegis 163

175 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Enrichment foods Daily supplements Is the den indoors or outdoors and is it on exhibit or of exhibit Is the den a pen or diggable area? Dickerson Park Zoo Flamingo Gardens carrots, dog food, omnivore, bell pepper, crawdads, live minnows thiamin E paste Both Pen live fish, birds they catch, ice blocks with fish, and meat Oasis vitamin Outdoors Pen, diggable Lee Richardson Zoo rib bones, live fish, raw hides vitamin E, vitamin B1, thiamine Indoors Diggable Little Rock Zoo various fish, hard boiled eggs, carrots, pumpkins none Indoors Pen Minnesota Zoo live minnows, chicks Omega 3 and 6 tablets Indoors Oakland Zoo hard-boiled eggs, chicks, bones, mealworms, crickets, goldfish, mice, fruits/vegetables Mazuri thiamin-e paste, 0.1 ml per kg fish fed Indoors Pen area 164

176 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Number of dens Nest materials provided Are the nesting materials seasonal? If seasonal, how do the nesting materials differ? Dickerson Park Zoo 2 pine shavings Flamingo Gardens 6 leaves that fall in exhibit, especially from the water Lee Richardson Zoo 1 hay Yes more in winter months if pregnancy suspected Little Rock Zoo 1 grass hay Minnesota Zoo 2 straw Yes bedding Oakland Zoo 1 + bed areas straw and blankets/towels No 165

177 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Dickerson Park Zoo Medium (> 1/2 up to 1 hr) [2-3 times daily] No No Flamingo Gardens Medium (> 1/2 up to 1 hr) [2-3 times daily] No, but otters are on exhibit during cleaning No Lee Richardson Zoo Yes Both Little Rock Zoo Medium (> 1/2 up to 1 hr) [2-3 times daily] No No Minnesota Zoo Medium (> 1/2 up to 1 hr) No Yes Oakland Zoo High (> 1 hr) No No 166

178 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Palm Beach Zoo at Dreher Park 9 x2 Florida Wild 11 x2 Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) 13, 15 Louisiana Wild 2 Sedgwick County Zoo Louisiana Wild

179 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of female Captive born or Wild caught Male and female together year-round or seasonally Palm Beach Zoo at Dreher Park Lowry (Tampa Bay, FL), Captive Year-round Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) Louisiana Wild Seasonally Sedgwick County Zoo Louisiana Wild Year-round 168

180 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Foods included in regular daily diet Enrichment foods Palm Beach Zoo at Dreher Park Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) Nebraska canine diet, fish, hardboiled eggs on Fridays fish, Nebraska feline, AAA brand feline, dry ferret chow, grated carrot, hardboiled egg, knuckle bone, pinky rat, chicks, dog biscuit hard-boiled eggs, carrots yams cored apples with fish inside, ice blocks crayfish, shrimp, squid, clams, salmon, tuna, sardines, corn, broccoli, banana, apple, grapes, crickets, mealworms, dry kibble mix (ferret, dog, cat, trout pellets), fish sickles, raw eggs, emu or ostrich eggs (raw or scrambled), live chickens, mussels, whip cream, cheese whiz, live fish Sedgwick County Zoo feline diet, perch, trout live fish, crawdads 169

181 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Daily supplements Is the den indoors or outdoors and is it on exhibit or off exhibit Is the den a pen or diggable area? Number of dens Palm Beach Zoo at Dreher Park Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) cod liver oil, Zoovite, Thiamine E paste on fish Indoors 1/female mazuri 1/2 # bird vitamins, cod liver oil, Thiamine (B1), vitamin E. 3 indoors; 1 only for female smell Inside is concrete floor, outside both exhibit and off exhibit is diggable dirt. 3 in, 2 man-made burrows outside Sedgwick County Zoo none Indoors pen 2 170

182 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Nesting materials provided Is the nesting material seasonal? If seasonal, how do the nesting materials differ? Palm Beach Zoo at Dreher Park hay No Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) straw, Leaves, grasses, hay, mulch, shredded paper, toilet paper, paper toweling, phone books to shred, towels yes Ravenous for nesting material during pseudopregnancy Sedgwick County Zoo prairie hay yes thick in breeding season 171

183 Table D1cont d. Table showing the answers given to the North American river otter (NARO) survey provided by successful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Palm Beach Zoo at Dreher Park Pueblo Zoo (answers for older female; information is the same concerning the new, younger female added in 2013) Medium (> 1/2 up to 1 hr) Yes Yes Medium (> 1/2 up to 1 hr) No Yes Sedgwick County Zoo Yes Yes 172

184 Table D2. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Buffalo Zoo 7 Sedgwick County Zoo, MO Captive 7 x2 Connecticut's Beardsley Zoo 9 Missouri Captive 2 Dakota Zoo 9 Louisiana Wild 6 Fort Worth Zoo 6.58 Texas Wild 1.75 Hutchinson Zoo 18 Private Citizen Captive 8 173

185 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of female Captive born or Wild caught Male and female together year-round or seasonally Buffalo Zoo Pittsburgh Zoo & Aquarium Captive Year-round Connecticut's Beardsley Zoo Dickerson Park Zoo, Springfield, MO Captive Year-round Dakota Zoo Homosassa Springs Wildlife State Park, FL Captive Seasonal Fort Worth Zoo Florida Wild Year-round Hutchinson Zoo Sedgwick County Zoo, MO Captive Year-round 174

186 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Foods included in the regular daily diet Enrichment foods Buffalo Zoo Nebraska feline diet, fish (capelin usually) carrots, hard boiled eggs, day old chicks, whole mackerel, herring, beef rib bones, cat food, crickets, superworms Connecticut's Beardsley Zoo Nebraska feline diet, capelin, herring chicken, beef, deer meat, rabbit meat, rats, chicks, mice, pumpkins Dakota Zoo Meat, fish carrots, live fish Fort Worth Zoo Capelin, chunk meat, smelt, trout, canned tuna, blue crab chew hooves, clams, crayfish, crickets, fish goo, live minnows, mussels, night crawlers, superworms, cheerios, dog biscuits, apples, carrots, coconut, pumpkin Hutchinson Zoo Feline carnivore meat, Nebraska brand with ground Science Diet dog food, carrots, and cod liver oil. hardboiled eggs, trout, and smelt 175

187 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Daily supplements Is the den indoors or outdoors and is it on exhibit or of exhibit Is the den a pen or diggable area? Number of dens Buffalo Zoo National mink pellets dog houses indoors, logs outdoors Pen, diggable mulch piles 4 dog houses Connecticut's Beardsley Zoo thiamin E paste, fatty acid Indoors Pen 2 Dakota Zoo included in AAA brand carnivore meat Indoors nest box, outdoors diggable Diggable outside 1 indoors, 1 outdoors Fort Worth Zoo thiamin E paste Indoors exhibit land is dirt 1 Hutchinson Zoo cod liver oil 2 176

188 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Nesting materials provided Is the nesting material seasonal? If seasonal, how do the nesting materials differ? Buffalo Zoo timothy hay No Connecticut's Beardsley Zoo straw Dakota Zoo hay No Fort Worth Zoo wood wool, towels Yes none offered off season Hutchinson Zoo hay No 177

189 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Buffalo Zoo No No Connecticut's Beardsley Zoo Medium (> 1/2 up to 1 hr) [3 training sessions per day] Yes and No Yes Dakota Zoo Low (< 1/2 hr) No No Fort Worth Zoo Medium (> 1/2 up to 1 hr) No Hutchinson Zoo Medium (> 1/2 up to 1 hr) No No 178

190 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Kansas City Zoo 4 John Ball Zoo, Grand Rapids, MI Captive 5 Lincoln Children s Zoo 8 Manhattan, NY Captive 12 Lincoln Park Zoo 7 Florida Wild 7 North Carolina Zoological Park 5 John's Island, SC Wild 9.5 Oregon Zoo 3 Star City, AR Wild 3 179

191 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of female Captive born or Wild caught Male and female together year-round or seasonally Kansas City Zoo St. Louis Zoo, ST. Louis, MO Captive Seasonal Lincoln Children s Zoo Louisiana Wild Both Lincoln Park Zoo Racine Zoo, Racine, WI Captive Year-round North Carolina Zoological Park Florida Wild Year-round Oregon Zoo Oregon-Pacific Northwest Wild Year-round 180

192 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Foods included in the regular daily diet Enrichment foods Kansas City Zoo capelin, dog chow, Natural Balance meat tuna, salmon, hardboiled eggs, apples, carrots, pig ears, minnows, crayfish, shrimp Lincoln Children s Zoo fish, horsemeat Lincoln Park Zoo Natural Balance meat, fish meaty cow bone North Carolina Zoological Park Oregon Zoo mackerel, capelin, lake smelt feline diet, ground polar bear chow, misc fish (herring, trout, salmon, capelin, smelt) hardboiled egg, carrots, tomato, crawfish, tilapia, sardines, herring, shrimp, sweet potato, frogs, mice, chicks chicks, mice, crab, crawdads, sweet potato, rawhide, pig ear, seaweed, crickets, mealworms, goldfish, rosehips, pumpkins (play with more, with a few bites), earthworms, salmon eggs, clams, hardboiled eggs, monkey chow, peanuts or pine nuts, krill, grapes, blueberries, apple, melon, orange, corn on the cob, chicken necks 181

193 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Daily supplements Is the den indoors or outdoors and is it on exhibit or of exhibit Is the den a pen or diggable area? Number of dens Kansas City Zoo vitamin B1 and vitamin E daily Moveable no 2 Lincoln Children s Zoo Outdoors n 3 Lincoln Park Zoo thiamine E paste Indoors no 1 North Carolina Zoological Park thiamin E Paste Indoors and outdoors Diggable outdoors, straw interior 5 (3 indoors/2 outdoors) Oregon Zoo Mazuri Vitamin Supplement daily Indoors in holding no one made specially, dog kennels 182

194 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Nesting materials provided Is the nesting material seasonal? If seasonal, how do the nesting materials differ? Kansas City Zoo mostly hay, sometime wood wool Yes Doesn't get hay in summer very often Lincoln Children s Zoo straw, towels No Lincoln Park Zoo straw, towels Yes North Carolina Zoological Park straw, timothy hay, leaves Yes towels, straw during winter straw, towels, hammock, sheets, blankets, fleece, leaves, pine needles, timothy, cut grass Oregon Zoo carwash strips, woodwool No 183

195 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Kansas City Zoo Medium (> 1/2 up to 1 hr) No No Lincoln Children s Zoo Low (< 1/2 hr) Yes No Lincoln Park Zoo High (> 1 hr) No North Carolina Zoological Park High (> 1 hr) Yes No Oregon Zoo High (> 1 hr) Yes Yes 184

196 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Pueblo Zoo 11, 13 Louisiana Wild 9 Roosevelt Park Zoo 6 Florida Wild 6 Rosamond Gifford Zoo 5 Dickerson Park Zoo, Springfield, MO Captive 2, 8 Seneca Park Zoo 21.5 Louisiana Wild

197 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of female Captive born or Wild caught Male and female together year-round or seasonally Pueblo Zoo Michigan Wild Seasonal Roosevelt Park Zoo Florida Captive and Wild Year-round Rosamond Gifford Zoo Florida, New York Wild Year-round Seneca Park Zoo Kentucky Wild Year-round 186

198 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Foods included in the regular daily diet Enrichment foods Pueblo Zoo fish, Nebraska feline, AAA brand feline, dry ferret chow, grated carrot, hardboiled egg, knuckle bone, pinky rat, chicks, dog biscuit crayfish, shrimp, squid, clams, salmon, tuna, sardines, corn, broccoli, banana, apple, grapes, crickets, mealworms, dry kibble mix (ferret, dog, cat, trout pellets), fishsicles, raw eggs, emu or ostrich eggs (raw or scrambled), live chickens, mussels, whip cream, cheese whiz, live fish Roosevelt Park Zoo Capelin, smelt, AAA Brand Feline Diet mice, minnows, chicks, fruit/veggies Rosamond Gifford Zoo Toronto carnivore diet (horsemeat) and whole fish (mostly smelt and/or capelin) live fish, crayfish, fruit, ice blocks with frozen fruit or fish Seneca Park Zoo meat, fish (Recently changed from Natural Balance to Cargill) live fish occasionally 187

199 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Daily supplements Is the den indoors or outdoors and is it on exhibit or of exhibit Is the den a pen or diggable area? Number of dens Pueblo Zoo Mazuri 1/2 # bird vitamins, cod liver oil, vitamin B1, vitamin E. 3; 1 only for female smell Inside is concrete floor, outside both exhibit and off exhibit is diggable dirt. 3 indoors, 2 manmade burrows outdoors Roosevelt Park Zoo none Indoor Pen 1 Rosamond Gifford Zoo In holdings 2/female Seneca Park Zoo Outdoors 3 188

200 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Nesting materials provided Is the nesting material seasonal? If seasonal, how do the nesting materials differ? Pueblo Zoo straw, leaves, grasses, hay, mulch, shredded paper, toilet paper, paper toweling, phone books to shred, towels Yes Ravenous for nesting material during pseudopregnancy Roosevelt Park Zoo blankets No Rosamond Gifford Zoo straw, grass hay No Seneca Park Zoo 189

201 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Pueblo Zoo Medium (> 1/2 up to 1 hr) No Yes Roosevelt Park Zoo High (> 1 hr) No Yes Rosamond Gifford Zoo High (> 1 hr) Yes No Seneca Park Zoo High (> 1 hr) Yes 190

202 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Male age Origin of male Captive born or Wild caught Female age Texas State Aquarium 2, 4, 8 Unknown, Unknown, Louisiana Wild 9, 15 Trevor Zoo 5 Buttonwood Zoo, New Bedford, MA Captive 3.58 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Origin of female Captive born or Wild caught Male and female together year-round or seasonally Texas State Aquarium Louisiana, Texas Wild Year-round Trevor Zoo Louisiana Wild Year-round 191

203 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Foods included in the regular daily diet Enrichment foods Texas State Aquarium fish (capelin), Natural Balance feline diet, fruits and vegetables All fruits and vegetables, Jell-O, fish pops, ice cube filled with food frozen Trevor Zoo mainly Nebraska feline diet fish, mice, chicks Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Daily supplements Is the den indoors or outdoors and is it on exhibit or of exhibit Is the den a pen or diggable area? Number of dens Texas State Aquarium Mazuri mammal, no vitamin A, Linatone Trevor Zoo Clovite and Linatone Outdoors pen around box 2 192

204 Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Nesting materials provided Is the nesting material seasonal? If seasonal, how do the nesting materials differ? Texas State Aquarium Trevor Zoo hay, but the otters bring in leaves No Table D2 cont d. Table showing the answers given to the North American river otter (NARO) survey provided by unsuccessful captive breeding institutions. Results collected from Association of Zoos and Aquariums accredited institutions from Name of Institution Amount of keeper- NARO interaction Is there direct contact? Does the respondent think their keepers have an impact on successful captive breeding? Texas State Aquarium High (> 1 hr) Yes Trevor Zoo Low (< 1/2 hr) Yes No 193

205 Appendix E. Pedigree Analysis I. Pedigree Legend II. Successful Captive Breeding Institutions (n = 15) III. Unsuccessful Captive Breeding Institutions (n = 16) 194

206 I. Pedigree legend Institution Name Institution Location 195

207 II. Successful Captive Breeding Institutions (n = 15) Alexandria Zoological Park Binghamton Zoo at Ross Park Brookgreen Gardens Cincinnati Zoo and Botanical Gardens Columbus Zoo and Aquarium Denver Zoo Dickerson Park Zoo Flamingo Gardens Lee Richardson Zoo Little Rock Zoo Minnesota Zoo Oakland Zoo Palm Beach Zoo at Dreher Park Pueblo Zoo Sedgwick County Zoo 196

208 Alexandria Zoological Park Alexandria, VA Binghamton Zoo at Ross Park Binghamton, NY 197

209 Brookgreen Gardens Zoo Pawleys Island, SC Cincinnati Zoo and Botanical Gardens Cincinnati, OH 198

210 Columbus Zoo and Aquarium Powell, OH Denver Zoological Gardens Denver, CO 199

211 Dickerson Park Zoo Springfield, MO Flamingo Gardens Davie, FL 200

212 Lee Richardson Zoo Garden City, KS Little Rock Zoological Gardens Little Rock, AR 201

213 Minnesota Zoological Gardens Apple Valley, MN 202

214 Oakland Zoo Oakland, CA 203

215 Palm Beach Zoo at Dreher Park West Palm Beach, FL Pueblo Zoo Pueblo, CO 204

216 Sedgwick County Zoo Wichita, KS 205

217 Unsuccessful Captive Breeding Institutions (n = 16) Buffalo Zoo Connecticut s Beardsley Zoo Dakota Zoo Folsom Children s Zoo Fort Worth Zoo Hutchinson Zoo Kansas City Zoo Lincoln Park Zoo North Carolina Zoological Park Oregon Zoo Pueblo Zoo Roosevelt Park Zoo Rosamond Gifford Zoo Seneca Park Zoo Texas State Aquarium Trevor Zoo 206

218 Buffalo Zoological Gardens Buffalo, NY Connecticut s Beardsley Zoo Bridgeport, CT 207

219 Dakota Zoo Bismarck, ND Folsom Children s Zoo Lincoln, NE 208

220 Fort Worth Zoological Park Fort Worth, TX Hutchinson Zoo Hutchinson, KS 209

221 Kansas City Zoo Kansas City, MO 210

222 211

223 Lincoln Park Zoological Park Chicago, IL North Carolina Zoological Park Asheboro, NC 212

224 Oregon Zoo Portland, OR Pueblo Zoo Pueblo, CO Roosevelt Park Zoo Minot, ND 213

225 Rosamond Gifford Zoo at Burnet Park Syracuse, NY Seneca Park Zoo Rochester, NY 214

226 Texas State Aquarium Corpus Christi, TX Trevor Zoo Millbrook, NY 215