A. Interim Report Form For Research Projects Funded through the National Science, Technology and Innovation Plan (NSTIP)

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1. PROJECT INFORMATION Project Code Project Title A. Interim Report Form For Research Projects Funded through the National Science, Technology and Innovation Plan (NSTIP) 11-ENV1918-02 Living on the edge: unravelling how Arabian Oryx (Oryx leucoryx) survive in the extreme deserts of the Empty Quarter (Rub Al-Khali) and Mahazat as-sayd protected area, Saudi Arabia. Dr. Abdulaziz N. Alagaili King Saud University Environment Nov. 2013 Nov. 2015 Nov. 2013 Dec. 2014 Principal Investigator Institution Strategic Technology Area Project Period (Starting Month/Year Ending Month/Year) Reporting Period (Starting Month/Year Ending Month/Year) Project Budget (SR) SR 1,990,200 Project Summary (max. 300 words) The Arabian Oryx is an iconic species of the deserts of the Middle East and is one of the success stories for conservation biology, being brought back from the brink of extinction. Despite the prominence of this animal from a conservation aspect we know relatively little about the survival strategies practiced in the wild which includes the inhospitable Empty Quarter (EQ). We have used state of the art research technologies the use of satellite collars (global positioning system) and associated thermal body temperature loggers that relay to the collar to monitor the movement and body temperature patterns of two populations of Arabian oryx in the extreme challenging environment of the Uruq Bani Ma arid (UBM) within the EQ and the Mahazat as-sayd (MS), a fenced protected area. The Arabian oryx has been kept in the protected area for more than twenty years and as a consequence, the population being monitored in MS will be used as a control against which to measure a number of variables that we will also measure in the population in the EQ. The Global Positioning System equipment has been used to investigate movement patterns in the two localities so that we can investigate whether the population in the harsher region (EQ) is more nomadic compared to the population in the less harsh region (MS). Furthermore, we are currently investigating the daily energy expenditure (DDE) of the Arabian oryx to ascertain what the food, water and energy requirements of these animals are and whether there are differences in metabolic requirements between seasons (summer and winter). For this latter work we are using the doubly labelled water technique. Finally, we are using actiwatches to monitor if there are seasonal changes in the sleep patterns of the Arabian oryx. The diversity of specialist scientists from both within Saudi Arabia and from other parts of the world will ensure shared knowledge transfer that is essential to the success of the study. At the end of February we will have collected an entire calendar year on individual animal movement, body temperature data patterns as well as energy budgets. In year 2 (2015), we plan to continue the winter season for the doubly labelled water, analyze the collected data, and write up three primary research papers on sleep, energetics and Arabian oryx home range and movement patterns, as well as a compound paper comparing the movement patterns and physiological variables in the two localities. 1

2. PROJECT PROGRESS Describe the objectives, phases, and tasks outlined in the original proposal and indicate the status of each task. This information can be inserted from Form RE-D1-3 of the original proposal, as long as the information contained therein was comprehensive. Otherwise, pertinent information from the original proposal can be entered in the table below (limit table to one page). Objectives Phases Tasks Movement of Arabian Oryx in the Mahazat as-sayd and Uruq Bani Ma arid protected areas of the Arabian Peninsula over a one year period to investigate if Arabian Oryx possess distinct home ranges or whether they are nomadic. Obtaining hourly body temperature (T b) measurements using implanted temperature transmitters that will be placed in the abdominal cavity (one year). Determining daily energy expenditure DEE and water turnover WTO of Oryx during both summer and winter in the protected area of Mahazat as-sayd. Investigating the daily activity patterns of Oryx using Actiwatch activity event monitors to construct a daily pattern of activity in relation to movement and body temperature exhibited in the Mahazat as-sayd. One, Two, Three, Four One, Two, Three, Four Two, Three, Four One, Three, Four Deployment of the collars, downloading and processing data for analysis, analysis Implant of body temperature transmitters, downloading and processing data for analysis, analysis Injections with doubly labelled water (summer/winter), lab analysis Implant of Actiwatch, downloading and processing data for analysis, analysis (*) Give details below if modified or discontinued. Status (Completed, Ongoing, Planned, Modified *, Discontinued*) Ongoing 70% Ongoing 70% Ongoing 50% Ongoing 80% Percentage of achievement 2

Twenty Arabian oryx were fitted with iridium satellite collars and abdomen body temperature sensors during February 2014. In March 2014, animals were released in Mahazat as-sayd (10 animals) and Uruq Bani Ma arid (UBM) protected areas (10 animals). Within a month of the release two Arabian oryx were killed by hunters in the UBM, an incident that confirmed illegal poaching of oryx in that unfenced protected area that has happened over the past 20 years. As a consequence, the Saudi Wildlife Authority increased their activities to keep the animals within the core area of UBM, which in turn affected the free roaming of these animals. The use of these collars allowed us to bring back those animals leaving the protected area to nearby towns (an individual moved over 70 km within few hours). We also were pleased to be able to observe other un-collared Arabian oryx joining the collared ones. Our team highly recommends the use of a few satellite collars during every future release event taking place within UBM. Body temperatures have been recorded every ten minutes for close on one year now. At the beginning of March we should have a full calendar year for at least 6 animals from the MS and 8 for the UBM. These body temperature measurements and the associated ambient temperatures recorded from the collar should enable us to determine the extremes of ambient temperatures that the oryx experience and the concomitant body temperatures experienced by the animals under such environmental conditions. A glance at the raw data from one Arabian oryx occurring in the UBM reveals that during the summer months, the Arabian oryx experiences a greater range of body temperature than those experienced in the winter. The Arabian oryx thus employs behavioural heterothermy by potentially heating up in the day and unloading heat during the relatively cooler evenings, in much the same way as the camel. We shall, in March, analyze the body temperature profiles of the two populations in line with the ambient temperatures experienced in the two areas. A glance at Figure 1 reveals that during winter the body temperature extremes are less than those recorded for the summer period. Unfortunately, we experienced a 40% failure rate in the deployment of the body temperature thermistors shortly after the release of the Arabian oryx into the two study areas. Although 40% of loggers failed, the data are still good enough to determine the range of core body temperatures experienced by oryx throughout the year. We have been fortunate to be able to track 18 Arabian oryx (10 at MS, 8 at EQ) for a period of close on a calendar year at two areas that have historically supported this animal (Fig. 2, 3,4,5). We plan to investigate sex differences in movement patterns as well as the home range of each individual at the UBM and MS. We plan to investigate seasonal changes in movement of the two populations with a view to assessing if the Arabian oryx in the harsher habitat (UBM) exhibit a different pattern of movement to those in the less arid habitat (MS). We have been fortunate to be able to track 18 Arabian oryx for a period of close on a calendar year at two areas that have historically supported this animal (Fig. 2, 3,4,5). We plan to investigate sex differences in movement patterns as well as the home range of each individual at the UBM and MS. We plan to investigate seasonal changes in movement of the two populations with a view to assessing if the Arabian oryx in the harsher habitat (UBM) exhibit a different pattern of movement to those in the less arid habitat (MS). We plan to analyze these pattern movements in April, May and June. We will attempt to assess home range of individual Arabian oryx and the groups using minimum convex polygons and kernels. A total of ten Arabian oryx, five in January and five in June were used for investigating the activity patterns and sleep at MS. Following anaesthesia, two actiwatches were placed into subcutaneous pockets (one on the neck and one on the hindleg) of each Arabian oryx. The Arabian oryx were then released into a 2 x 2 km pen, where there was no provisioning of food, but the Arabian oryx were allowed access to water. Following 3 months of free ranging, the 3

Fig. 1: Body temperatures record for a female Arabian oryx at MS (10 months). Fig. 2: Movement of a female Arabian oryx at MS (10 months). Fig. 3: Movement of a male Arabian oryx at MS (10 months). 4

Fig. 4: Movement of a female Arabian oryx at UBM (10 months). Fig. 5: Movement of a male Arabian oryx at UBM (10 months). animals were immobilized and the implants removed. During the winter, the average daily inactivity or sleep was 4.87 hours. The average daily onset of the major sleep bout occurred at 3:16 am, with this bout having an average length of 124 minutes. Thus, during winter, the Arabian oryx displayed a diurnal activity pattern, with the vast majority (93%) of inactivity or sleep, occurring during the night. During summer the average daily inactivity or sleep was 6.03 hours. The average daily onset of the major sleep bout occurred at 12:48 pm with an average length of 119 minutes. Thus, when compared to winter, the summer daily activity and sleep 5

pattern of the Arabian oryx changed dramatically, becoming a crepuscular activity pattern, having an extra hour of sleep, with 65% of sleep occurring in the daylight period (Fig. 6). This seasonal change in the daily sleep pattern is a novel finding, not recorded in any other animal. While this portion of the study is near complete (data analysis ongoing), the results have led us to continue the sleep work by recording physiological sleep with implanted electrodes in 6 Arabian oryx (3 in winter, 3 in summer) to definitively determine when the Arabian oryx sleep in the different seasons. It appears that this change in sleep patterns would allow the Arabian oryx to overcome the problems of heat overload, and, as body temperature drops during sleep, providing them with a novel way to thermoregulate in the summer. Fig. 6: Average activity patterns and sleep of Arabian oryx To determine daily energy expenditure DEE and water turnover WTO in Arabian oryx during both summer and winter in the protected area of Mahazat as-syad (MS). We decided to 6

inject a total of 20 Arabian oryx with doubly labelled water. We selected the MS to do the work in because it is possible to collect the samples from the animals at this locality. In the UBM the constantly shifting sands makes this impossible. We hypothesized that the Arabian oryx will expend reduced energy, but will show a significantly higher water turnover during the summer compared to winter. These measurements will be related to the energy and foraging needs of wild Arabian oryx in order to determine how they are able to survive in extremely arid environments. To date we have injected 10 Arabian oryx with doubly labelled water (DLW) during August 2014 (summer) and are due to inject another 10 animals in February 2015 (winter). Prior to conducting the research we undertook a pilot study in a captive Arabian oryx at the King Khalid Wildlife Research Centre in order to determine the optimal dose of DLW, the equilibration time of isotope in the body of the animal, and also the elimination rate of the isotope. These aspects are necessary in order to use the DLW technique effectively. For this purpose, 10.0 ml of deuterated water (99.9% enriched 2 H water) was injected IV into a captive female Arabian oryx. A series of blood samples were collected at various time intervals afterwards in order to determine both the isotope equilibration rate - the time taken for isotope to mix up within the body of the animal - and the isotope elimination rates - the time taken for isotope to be eliminated from the body of the animal, both short and long term - (Fig.7,8). We plan to do undertake the winter component of the research in February, 2015 having already accomplished the summer component in August, 2014. The relative amounts of deuterium and heavy oxygen will be determined by isotopic ratio mass spectrometry and the necessary calculations undertaken to assess field metabolic rate and water flux, by examining the differential elimination rates of the two isotopes 700 KKWRC Blood Deuterium 600 500 Deut PPMs 400 300 200 100 0 19/04 00:00 20/04 00:00 21/04 00:00 22/04 00:00 23/04 00:00 Fig. 7: Isotope equilibration rate, short-term scale, to determine equilibration rates post dose and optimum times for initial sample 7

600 KKRRC Blood Deuterium Deut PPMs 500 400 300 200 100 0 01/05/2014 00:00 26/04/2014 00:00 21/04/2014 00:00 16/04/2014 00:00 16/05/2014 00:00 11/05/2014 00:00 06/05/2014 00:00 Fig. 8: Isotope elimination rate, long-term scale, to determine elimination rates and optimal times for final sampling post dose In conclusion, we are extremely happy with the progress of the research and we feet that all the objectives outlined in the original submission are being attained. The research has required a great deal of coordinated field research by a number of individuals. We are also extremely indebted to the Saudi Wildlife Authority for their continual support of the project in the form of manpower and logistics and most importantly, the donation of oryx. 8

3. PERSONNEL INVOLVEMENT Provide details addressing the contributions of each person in accomplishing the project tasks/objectives during the reporting period. Include personnel involved that are not supported by NSTIP award. Team Members Name Contribution Investigator Dr. Abdulaziz Alagaili Directing the project in all its aspects. Co-investigator/s Consultant/s Student/s (specify undergraduate, M.S. or Ph.D.) Research Staff (e.g., Postdoctoral Associate, Technician, Manager) Administrative Staff Others (provide title and role) 1)Prof. N. Bennett, 2)Prof. P. Manger, 3)Prof. J. Speakman, 4)Dr. D. Scantlebury, 5)Dr. O. Mohammed. None Due to the delay in providing funds, a graduate student could not be identified. Mr. Abdulrahman Alohali None Workers 1) Physiological analysis and coordinating with PI and other team members. 2) Actiwatch analysis. 3,4) Doubly labelled water field work and analysis. 5) Field work assistance and data collection. Data management Ground work, moving equipment, capturing animals, field work assistance. Describe any changes in personnel and/or their roles from the original proposal. Please also include individuals not included in your original proposal that provided significant contribution to the project to date. (max. 150 words) No changes in the project personnel except the involvement of Dr. Mads Bertelsen, a veterinary expert from Copenhagen Zoo who became the project vet. Dr. Bertelsen undertook the anaesthetic work and the implanting of the abdominal sensors. He has made several trips to Saudi Arabia to ensure the welfare of the animals, working on over 40 animals during the course of the 1 st year. Thankfully, none of the animals died during or post the surgery. Two Arabian oryx died a month post-surgery due to old age. 9

4. RESEARCH OUTPUTS Include information in the following categories that directly relates to your NSTIPfunded project. Include details, status (e.g., accepted, published, submitted, under preparation), and dates for each category that applies to your project. OUTPUTS: Patents: (List details on a separate sheet) Quantity Status Date Patent applications Patent registrations Licenses Other research commercialization activities Publications: (List on a separate sheet, the details of the publication, e.g., title, journal, impact factor with source such as Thomson Quantity Status Date Reuters ISI) Refereed publications 4 ongoing 2015/16 Non-refereed publications such as journal articles, reviews, conference papers, books and book chapters Presentations Quantity Status Date State the quantity and list & specify on a separate sheet whether they were conference talks, seminars, lectures, invited talks, etc., and whether they were institutional, regional, national or international. None up this point. Technical Outputs Quantity Status Date List below any technical outputs such as CDs, software programs, databases, algorithms, and measurement instruments. Service to the Research Community Quantity Status Date List any membership on national and international science committees, advisory boards, journal editorial boards, conference organizing committees, etc. 10

Impact on Policy Quantity Status Date List any contributions to development of research or clinical 1 Regulations to protect oryx against poaching are under guidelines, review protocols, consideration at Saudi 2015/16 indicators, membership on Wildlife Authority. The government advisory committees, project PI has been assigned by the president of SWA to commissioned governmental lead a team to prepare a agencies, meetings with policy strategic plan for future makers, etc. release/monitoring of Arabian oryx. Awards and Honors Quantity Status Date List any major awards, indicating their scale regional, national, international and honors such as academic chairs and endowed positions. Other Quantity Status Date List any other forms of research dissemination that is intended for non-scientific audiences (such as radio talks, newspaper articles, television appearances). 2 * Local TV and newspaper interviews before/after the release of Arabian oryx at the Empty Quarter. *SWA announced this project during international conventions. 2014 11

` 5. BROADER IMPACTS OF THE PROJECT Include information in the following categories that directly relates to your NSTIPfunded project. Include details and dates for each category that applies to your project. Teaching and Training Describe courses, classes, and workshops that were developed, and your role in the activity (for example, teacher, organizer, developer). - The project provided an opportunity to train Saudi Wildlife Authority personnel (researchers and rangers) on field techniques (capturing, anaesthesia process, monitoring anesthetized animals, release techniques, on ground monitoring after release, and online monitoring of animals through satellite. - A workshop regarding the project implemented techniques and outcome will be held at King Saud University. KSU president, SWA president, and KACST vice president will be invited for part of this workshop where the results will be presented. All three individuals have expressed a great interest in finding out about this project results, which shows a great interest in decision makers to such projects in this field. Infrastructure If you purchased equipment, describe how it adds to the capability of the institution and training of researchers outside the project. Indicate whether this equipment is available elsewhere in the institution and why its purchase was necessary for this project. The nature of this project is based on field work, thus most purchased equipment was attached/implanted in the studied animals, which were then released to the wild. It would be a challenging task to recover this equipment, especially the collars, but we will try to recover these collars if possible from animals released in MS and refurbish them to be used in future release activities. The project fund enabled us to secure basic equipment needed at most laboratories such as sensitive electronic scale, animal scales, and dissection tools amongst other items of small equipment. Collaborations Describe the institution, disciplinary focus, research expertise, and nature of any new or unforeseen partnerships that were developed during the project. Our research group at King Saud University focuses on biological and ecological studies on mammalian species that occur in Saudi Arabia. The current project is not only a continuation of a strong collaboration with colleagues from South Africa, but also has provided us the opportunity to expand our network to be working closely with other colleagues from United Kingdom and Denmark. Beside the fruitful data coming out of this project, the team has decided to proceed with future work aiming to study three other endangered Artiodactyl species found in Saudi Arabia (Arabian and sand gazelles, and the Nubian Ibex). Funding Describe how this research has led to additional funding or prospects for future funding awards and contracts for project investigators, students and staff. Please list the applications, dates, reference numbers, and amount awarded, if applicable. None. 12

Others Describe the benefits of your research to society that are not covered by the categories above. The Arabian oryx is an iconic species that captures the attention of people across the world, and specifically those living in the Arabian Peninsula. The establishment of SWA was a result of international efforts to save this species from extinction. The Royal Family of Saudi Arabia gave the Arabian oryx special care and attention by protecting them not only in protected areas and breeding centers but also on their private farms. This project, and hopefully future research plans, will provide more insights about the life history, behavior and physiology of the Arabian oryx that should be well received by the scientific and conservation communities across the world. Many colleagues who have learned about this ongoing project have asked to be informed about the preliminary results. The same attention and excitement was observed within the country and many decision makers (i.e. ministers and deputy ministers, government offices directors) already have asked for a report after the end of this project. We are really proud of this project and happy to see the support from colleagues and the government of Saudi Arabia. 6. BUDGET Category First Second year year Amount Awarded Amount Expended Amount Awarded Principal Investigator 72,000 0 72,000 0 Co-Investigator/s 180,000 0 180,000 0 Consultant/s Student/s (specify undergraduate, M.S., Ph.D.) Research Staff (specify 20,000 20,000 20,000 0 Postdoctoral Associate, Technician, Manager) Administrative Staff Major equipment (list items >100,000 SR below) 343,798 343,798 30,000 0 Other Equipment (list items >10,000 SR below) 178,202 178,202 78,197 0 Materials & Supplies 304,000 304,000 78,301 0 Travel (e.g., training, conference, field trips) Dissemination (e.g., publications, patents, workshops, public outreach) 100,000 100,000 240,000 0 0 0 50,000 0 13 Amount Expended

Other (specify any >1,000 SR) 0 0 10,000 0 Total 1,198,000 946,000 758,498 0 Please list equipment (>10,000 SR) purchased. In addition, provide brief descriptions of travel, dissemination, and other expenditures during the reporting period. If funding sources other than NSTIP have supported any aspect of the research program, please describe the funding amount and nature of support (financial, in-kind, facilities, etc.) Also, comment on any existing core or shared research facilities that you have used for the project to date. If category expenditures differ from budgeted amounts by more than 10%, please comment. (max. 150 words). - Satellite Collars + body temp loggers. - Travel category covered field work in the two protected areas. A training and conference attendance will follow up in the 2 nd year. 7. Describe any other concerns and comments related to the interim progress of the research project that were not covered in the sections above (max. 250 words). We have not been able to analyze in full the movement data or body temperature data currently because the project will only terminate at the end of March. In order to look at and present seasonal and sex related results we require the full calendar year of data to report accurately on these parameters measured. Similarly, we have not been able to report on the seasonal energy expenditure as winter samples will only be collected in February. The killing of two of our animals in the Empty Quarter has reduced our data set, but this was beyond our control. On a positive note, it has enabled us to report that releasing oryx into the EQ could pose some risk to the stock being released. Papers planned to be published out of this project: 1- Walk the walk: temporal and spatial patterns of movement in the Arabian Oryx. A manuscript will firstly be submitted to Journal of Animal Ecology (IF 4.7). 2- Too hot to handle: the body temperature rhythms of Arabian oryx in the extreme desert of the Empty Quarter. A manuscript will first be submitted to Biology Letters (IF 3.7) 3- From the cold of the night to the heat of the day: seasonal switching of sleep times in the Arabian oryx. A manuscript will first be submitted to PLoS Biology (IF=11.771). 4- Water and energy in extreme environments: How to Arabian Oryx survive with no water, extreme heat and little food? A manuscript will first be submitted to PLoS One (IF=3.73) Thanks 14