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Pakistan J. Zool., vol. 46(6), pp. 1679-1683, 2014. Parasitic Infection in an Imported Fish Fantail, a Variety of Goldfish, Carassius auratus L. in Pakistan Zafar Iqbal* and Husna Noreen Department of Zoology, University of the Punjab, Quaid-i-Azam Campus, Lahore-54590, Pakistan. Abstract.- The parasitic infection in an imported ornamental fish fantail, a variety of goldfish Carassius auratus purchased from pet shop in Lahore was studied. In total eight species of parasites belonging to protozoans, monogenean and digenean trematodes and crustacean were identified. Trichodina Ehrenberg 1831 infection on skin and fins was 20%, (mean intensity 9.5); Ichthyophthirius multifiliis Fouquest 1876 infection on gills was 43.3% and on fins 30% (mean intensity 20.72 and 29.25, respectively). There was no difference in the infection of fins and gills by I. multifiliis (χ² = 1.14, df = 11, p=0.05). Tetrahymena sp. infection was 10% (mean intensity 98). Gills showed 100% infection with Dactylogyrus Diesing 1850 (mean intensity 29.3), Gyrodactylus Von Nordmann 1832, infection on skin of fish was 70% (mean intensity 11.1). Metacercariae of an unidentified digenean were also found encysted on gills (infection 66.60%, mean intensity 20.4). Argulus foliaceus Muller, 1785 infection was 26.60% and mean intensity 6.0. A cyclopoid stage of Lernaea sp. was also observed on skin. Dactylogyrus infection was the highest which caused severe damage to gill filaments and secondary lamellae. This study points our attention to a serious issue of import of infected fish. Key words: Imported fantail, parasitic infection, Dactylogyrus sp. Gyrodactylus sp. INTRODUCTION Pet fish keeping is a hobby worldwide, mainly in developed countries (Olivier, 2003). The major part of aquarium fish industry is from freshwater fish sector (Helfman, 2007). Upto 2 million people worldwide are believed to keep marine aquaria (Wabnitz, 2003). FAO (2007) reported that world export of ornamental fish was almost US $ 283 million in 2006. South East Asian countries supply over 50% of the world's ornamental fish (Olivier, 2003). Many rural communities in South America, Africa and Asia earn their livelihood from this industry. Goldfish is the most popular and commonly kept aquarium fish in the world (Komiyama, et al., 2009). Ahilan et al. (2009) listed eleven varieties of goldfish including fantail. Twenty species of ornamental fishes are imported live into Pakistan from South East Asian countries (Ahmad, 1996). According to Andrews (2006) pet fish trade is of great benefit to many countries, yet it may have adverse effects as a result of spread of different pathogens through the introduction of nonnative species. The ectoparasites * Corresponding author: dr.zafariqbal.pu@gmail.com 0030-9923/2014/0006-1679 $ 8.00/0 Copyright 2014 Zoological Society of Pakistan are generally known to comprise the largest group of pathogens in warm water fish (Snieszko and Axelrod, 1971). Many studies reported the transmission of parasites and other pathogens through ornamental fishes worldwide. Some of these studies are from Germany, Australia, Korea, Norway, Sri Lanka, Brazil, Turkey, Pakistan (Moravec et al.,1999; Evans and Lester, 2001; Kim et al., 2002; Levsen et al., 2003; Thilakaratne et al, 2003; Pizza et al., 2006; Tavares-Dias et al., 2010; Iqbal et al., 2012a,b, 2013a,b; Iqbal and Sajjad, 2013; Iqbal and Mumtaz, 2013; Iqbal and Hussain, 2013; Haroon et al., 2014; Iqbal and Haroon, 2014; Iqbal and Rehman, 2014; Koyuncu, 2009; Kayis et al., 2013). In freshwater fishes, two monogenean parasites attack gills and skin and are narrowly host specific. There are 970 species of genus Dactylogyrus which infect gills of fishes (Gibson et al., 1996). Two species namely D. extensus and D. vastator are common gill parasites of cyprinids, while Gyrodactyus salaris and G. turnbulli infect skin and fins of fishes. The common protozoan parasites of freshwater fishes are Ichthyophthirius multifiliis, Chilodenella sp, Trichodina sp. Ichthyobodo necator and Tetrahymena sp. Most of the ectoparasitc protozoan produce serious diseases and may cause mortality when their infection is very

1680 Z. IQBAL AND H. NOREEN high. Among crustacean, two parasites of freshwater fishes are Argulus foliaceus and Lernaea cyprinacea which are cosmopolitan in distribution and infect almost all fishes (Bond, 2004; Noga, 2007; Rasouli et al., 2012). The aim of present study was to investigate and identify parasitic fauna of a freshwater ornamental fish fantail, imported to Pakistan. MATERIALS AND METHODS The experimental fish fantail was purchased from pet shop in Lahore from July to October 2013. The fishes were brought live in sterilized polyethylene bag and kept in glass aquarium in aerated water in the Lab. The fish were weighed, measured and examined for the presence of any parasites or lesions on the body of fish. The mucus from skin, gills, fins and operculum was scrapped with glass slide and then spread carefully with cover slip separately for examination of protozoan and monogenean gill and skin flukes, under compound microscope at x10 and x40 magnification. The fish gills were cut and examined under microscope for monogenean gill fluke. The specimens were preserved in 10% buffered formalin and 70% ethanol for storage before further studies. The crustacean parasites were collected from fins cleaned in saline and preserved in 70% alcohol. The fresh mount preparation technique was applied to observe the motile parasites (Post, 1987; Southgate, 1994). Parasites were identified according to Kabata (1985) and Lome and Dykova (1992). Chi-square test was applied to check difference in infection level on gills and fins. Photographs of slides of parasites were taken with digital microscope camera Digiprolabomed, USA. RESULTS Thirty fantail fish were examined for parasitic infection. The total length and body weight ranged from 7.5-12.5cm and 7.2-31.1g. Clinically, 10 fish were moribund which showed sluggish movement. Two fish had patches on the skin. Tips of fins were eroded and white spots were very clearly seen on them. The infected fishes were having 2 to 6 species of parasites attached to their fins, fin rays, gills, skin and head. The infection was 100% and mean intensity of infection was 91.2 in the whole fish sample. The parasites recorded were monogeneans such as Dactylogyrus sp. and Gyrodactylus sp. protozoans such as Trichodina sp., Tetrahymenea sp., I. multifiliis; an unidentified digenean metacercaria on gills; crustacean such as A. foliaceus and a cyclopoid stage of Lernaea sp. A total of 2737 parasites were recovered. The species wise prevalence, mean intensity and site of infection in given in Table I. Dactylogyrus infection on gills was 100% and it caused damage to gill epithelium and erosion of gill filaments. Gyrodactylus sp. had high infection (70%) but low mean intensity (11.1%). Tetrahymenea showed low infection (10%) but very high mean intensity (98). Trichodina infection was 20% and mean intensity 9.5. High mean intensity of infection (20.72) of I. multifiliis on gills is serious pathological condition for the fish. This has resulted in deformation of respiratory surface. Fin infection by I. multifillis was 30% and mean intensity 29.25. No significant difference was observed in infection of gills and fins by I.multifillis (χ² = 1.14, df = 11, p=0.05). Infection of encysted metacercaria of digenean on gill filament was high (66.60 %, mean intensity 20.4). The gill filaments became swollen. Whereas, Argulus foliaceus showed 26.60% infection and mean intensity was 6.0. Only one Lernaea was found on skin of fish. DISCUSSION The most common parasite on gills with highest infection was Dactylogyrus sp. It caused destruction of gill epithelium and disruption of tissues of gill filaments. The structure of capillaries are damaged and blood supply become poor, this affects respiratory function. This parasite had been reported worldwide but is most common in carps (Nematollahi et al., 2013). Gryodactylus sp. showed lower infection than Dactylogyrus sp. and seems less pathogenic than Dactylogyrus. However, the damage it causes to skin, scales and epithelium at the point of attachment on host is prominent because it form foci for entrance of secondary pathogens. Monogenean are host specific, have direct life cycle

PARASITIC INFECTION IN FANTAIL AN ORNAMENTAL FISH 1681 Table I.- Parasites recovered from Fantail, Carassius auratus. Parasite Infected fish Prevalence (%) Site of infection Total parasites Range of parasites Mean intensity Dactylogyrus sp. 30 100 Gills 879 5-72 29.3 Gyrodactylus sp. 21 70 Skins/fins 233 1-30 11.1 Trichodina sp. 6 20 Skins/fins 57 2-25 9.5 I. multifiliis 22 43 Gills 456 1-160 20.72 I. multifiliis 12 30 Fins 351 1-275 29.25 Tetrahymena sp. 3 10 Skins/fins/gills 294 65-120 98.0 Digenean metacercariae? 20 66.6 Gills 408 2-56 20.4 A. foliaceus 8 26.6 Fins 48 1-17 6.0 Lernaea sp. 1 3.3 Skin 1 1.0 1.0 2737 91,2 and their transmission is enhanced in poorly managed fish ponds (Thilakaratne et al., 2003; Woo, 2006). The results of the present study are comparable to Thilakaratne et al. (2003), Chand et al. (2011) and Kayis et al. (2013), who reported clear hemorrhagic areas on the skin of C. auratus infested with Gyrodactylus sp. and destruction of gill of fish infested with Dactylogyrus sp. High infection of encysted metacercariae of digenean on gills filaments as observed here may also prove to be fatal to fish too. Infection of I. multifiliis on skin and fins results in irritation, detachment of epidermis and mucus production, whereas gill infection cause gill damage and respiratory stress (Tavares-Dias et al., 2010). In heavy infection 100% mortality can occur. Infection by I. mulltifiliis cause white spot disease in fishes. This parasite is low host specific and widely distributed in freshwater fishes (Thilakaratne, et al., 2003; Tavares-Dias et al., 2010; Chanda et al, 2011; Iqbal et al., 2013b). Trichodina sp. occur in large numbers on fish under stressed conditions in ponds due to poor water quality and overcrowding, these parasites can proliferate massively and become serious ectoparasites (Lom and Dykova, 1992). Trichodina sp. has also been reported by Iqbal and Hussain (2013) in shubunkin C. auratus. High mean intensity of infection of Tetrahymena sp. was recorded from skin of the fish. Tetrahymena sp. infection has also been reported from six ornamental fishes viz., Poecilia reticulate, C. auratus, Poecilia sphenops, Puntius sp. Pterophyllum scalare, C. carpio from Sri Lanka (Thilakaratine et al., 2003) and from Carnegiella strigata in Brazil (Tavares- Dias et al., 2010) and in guppy, P. reticulata (Leibowitz and Zilberg, 2009). Tetrahymena infection is also known as guppy disease. Low infection of A. foliaceus was observed in fantail. However, it is potentially a serious pathogen of commercial fish (Iqbal et al., 2013a). High infection up to 800-1000 parasites per fish may cause mortality in C. carpio and other carps (Pekmezic et al., 2009). One Lernaea sp. was found on fish. This is the infective stage of parasite. Lernaea infection is serious problem in culturable and ornamental fishes (Iqbal et al., 2012a; Iqbal and Haroon (2014). This study has highlighted the issue of introduction of parasites with ornamental fishes imported into Pakistan. This unchecked import of diseased fish must be strictly regulated and controlled so as to safeguard our biodiversity and ecosystem. AKNOWLEDGEMENTS We are grateful to University of the Punjab, Lahore for providing funds for this study. The Conflict of interest declaration There is no conflict of interest or otherwise. REFERENCES AHILAN, B., FELIX, N. AND JAMESSON, J. D., 2009. Goldfish. Daya Publishing House, Delhi, India. AHMAD, N., 1996. Extraction, Exploration and Demand

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PARASITIC INFECTION IN FANTAIL AN ORNAMENTAL FISH 1683 RASOULI, S.A., NEKUIFARD, D., AZADIKHAH, H., AHARI, A., ANVAR, A., KHODADADI, A. AND GHASEMI, A., 2011. Ectoparasitic infection of Carassius auratus in water resources of West Azerbijan. Iran. Iran. J. Fish. Sci., 4: 156-164. SNIESZKO, S.F. AND AXELROD, R., 1971. Diseases of fishes. TFH Publication, New Jersey, USA. SOUTHGATE, P., 1994. Laboratory diagnosis of fish disease. Practice, 9:252-255. TAVARES-DIAS, M., LEMOS, G.R.J. AND MARTIN, L.M., 2010. Parasitic fauna of eight species of ornamental freshwater fish species from the middle Negro River in the Brazilian Amazoan Region. Rev. Bras. Parasitol. Vet. Jabotical, 19: 29-33. THILAKARATNE, I.D., RAJAPAKSHA, G., HEWAKOPARA, A., RAJAPAKSE, R.P. AND FAIZAL, A.C., 2003. Parasitic infections in freshwater ornamental fish in Sri Lanka. Dis. Aquat. Org., 54: 157-162. WABNITZ, C., 2003. From ocean to aquarium: The global trade in marine ornamental species (No. 17). UNEP / Earth print. WOO, P.T.K., 2006. Fish diseases and disorders. 1: Protozoan and metazoans, 2 nd ed., CABI Publication. Cambridge, UK. pp.791 (Received 15 May 2014, revised 7 August 2014)