ARGULOSIS, IN SOME ORNAMENTAL FISHES IMPORTED TO LAHORE, PAKISTAN

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Available online at http://scik.org European Journal of Veterinary Medicine, 2 (2013), No. 3, 171-178 ISSN 2051-297X ARGULOSIS, IN SOME ORNAMENTAL FISHES IMPORTED TO LAHORE, PAKISTAN ZAFAR IQBAL *, RABIA MUMTAZ AND RAMSHA SAJJAD Department of Zoology, University of the Punjab, Quaid-E- Campus, Lahore. P. code. 54590. Pakistan Abstract: Three imported ornamental fishes; black moor and shubunkin two varieties of goldfish, Carassius auratus L. and the third fish koi, a variety of Cyprinus carpio L. were obtained from pet shops in Lahore, Pakistan from August to October 2011. These fishes were examined for parasitic infection, argulosis. Black moor (n=24) showed 12.50% infection (mean intensity 4.5). Koi (n=22) had 13.63% infection (mean intensity 8.66). No infection was recorded from shubunkin (n=18). The causative agent, Argulus foliaceus L. a crustacean parasite, was attached on the head, fins and skin of black moor and Koi respectively. The infected fishes were listless, had faded skin color, eroded scales and excessive mucus secretion was evident. The lesions were present at the point of attachment of parasite on the skin of fish. The infection level was low (<14%) in both black moor and koi. These findings show a typical example of introduction of potentially pathogenic parasite such as, A. foliaceus from one geographical location to another along with its host. This situation may be alarming as the unchecked import of infected and diseased ornamental fishes may become threat to local aquatic ecosystem. So, the strict regulations on import of live, healthy and disease free fish may be implemented by the authorities. Key words: Argulus foliaceus, black moor, Koi, infection, threat. Introduction Argulus species are ectoparasites of freshwater fishes that are distributed worldwide in Europe, North America, Asia, Australia except in Antartica [2.16]. Argulus foliaceus has low host specificity and cause mortality in fishes [26, 27]. This parasite attach on various sites on fish, such as oral cavity, skin, gills and fins and result in debility, skin lesions, flicking of fins. Even the infected fish rub against the walls of aquarium [18, 14]. * Corresponding author Received January 22, 2013 171

172 ZAFAR IQBAL, RABIA MUMTAZ AND RAMSHA SAJJAD Argulus spp. produce large number of eggs in many clutches in one breeding season. The parasite has potential to reach high density quickly and spread throughout the water body. Low infection may leads to epizootics in densely held fish population, which causes large mortalities. Fish kill due to argulosis has been reported from fish hatcheries and nurseries, rearing ponds, fishing lakes, in home and public aquariums [12, 13, 9, 23,15]. The ornamental fishes such as goldfish, Koi carp, platy, Xiphophorus maculates; Oscar, Artronotus ocellatus [ 21, 25, 26, 1] have also been reported to have Argulus infection. Argulus japonicus is considered to be distributed in the world through the trade of carp and ornamental fish such as goldfish. Both A. japonicus and A. foliaceus can potentially survive off-host periods in water transport for 11-14days as adults but their survival is shorter as larvae [3, 27]. The aim of this study was to observe parasitic infection argulosis in three ornamental fishes, black moor and shubunkin two varieties of goldfish and koi carp imported from Thailand to Lahore as healthy and diseased free stock. Materials and Methods Three ornamental fishes, black moor, koi and shubunkin were obtained from pet shop in Lahore, Pakistan from April to October 2011. The fishes were brought live in sterile polyethylene bags in aerated aquarium water to the Fish Disease and Health Management Lab. The fishes were kept in 40 liters glass aquarium containing aerated water till examination within 2-3 hours. Fishes were weighed and measured. The body surface, fins and gills were examined for parasitic infection, any lesion or any other clinical sign. The Argulus parasites were collected and preserved in 70% alcohol. Some parasites were fixed in glacial acetic acid and preserved in glycerine-gel under cover slip according to [17]. Argulus foliaceus were identified according to [4, 3]. The parasites were measured with eye piece graticule fitted in microscope and photographed. Results and Discussion The causative agent of argulosis was identified as Argulus foliaceus, commonly called fish lice. The dimension of the parasite was 1.79-2.80 x 2.84-4.16 mm (Fig.2). Twenty four black moor were studied for argulosis. The fish had low prevalence (12.50%) (Table 1) the mean intensity was 4.66. The infected fishes were listless, with faded skin color and lesions on body (Fig.1). A lesion measuring 2.3x2.6 cm was prominent near caudal peduncle of one fish. At the point of attachment the scales were eroded and epidermis was exposed

ARGULOSIS, IN SOME ORNAMENTAL FISHES IMPORTED TO LAHORE, PAKISTAN 173 indicating old infection. The parasites attachment on skin was higher (78.57%) compared to all the fins (14.29%) and head (7.14%) (Table 2). Appearance of inflammation and secretion of mucus was very evident and common in infected fishes. Twenty two Koi carps were examined, and the fish showed low prevalence (13.63%). But the mean intensity was high (8.66). Argulus foliaceus were attached on the body, fins and head of the fish. Skin harboured higher number of parasites (61.53%) compared to all the fins (26.92%) and head (11.53%) (Table 2). The parasites were observed escaping from the fish and moving into petri dish containing tap water. The infected fishes had superficial lesions on the skin. Excessive secretion of mucus at the point of attachment of parasite was evident. Even the scales were eroded. No infection was observed in shubunkin. The infection level of argulosis was low in both fish species (<14) but the mean intensity of infection was higher in koi carp as compared to black moor. Low infection (1-2 parasites per fish) has no noticeable impact on fish, unless fish is small [18]. High parasite burden (800-1000 parasites/fish) in pond reared C. carpio, resulted 100% prevalence and high mortalities [15]. In the present study, mildly infected fishes showed mucus secretion, small hemorrhages and localized inflammation at the site of attachment of the parasite. In heavy infection the fish become lethargic, stop feeding and skin pigmentation is reduced. Attachment of A. foliaceus causes lesions on skin of fish, which are susceptible to secondary, bacterial and fungal infections [7]. Even Aeromonas salmonicida and Saprolegnia spp. infection on the damaged skin of fish is often concurrent with Argulus spp. infection [10]. The attachment of Argulus sp. on fish results skin and gills irritation due to insertion of pre-oral sting into the host and sucking body fluid with proboscis like mouth [18]. Argulosis is threat to commercial fisheries; both in warm and cold waters and Argulus spp. represent potentially a serious pathogen [8, 11, 2]. Infection by Argulus has been reported in tilapia, Oreochromis niloticus [19], rainbow trout, Oncorhynchus mykiss and C. carrpio and goldfish [12, 20, 1, 22, 15], rudd, Scardinius erythrophthalmus [14]. Argulus inducus, A. japonicus and Argulus siamenses have been reported to cause mortality in major and Chinese carps [3, 13, 9, 2]. Argulus foliaceus and Argulus bengalensis have been reported to infect 11 freshwater fishes in Bangladesh [6, 5]. The low host specificity of fish lice make it more dangerous. So, it may infect any freshwater fish. [23] pointed out risk factors associated with Argulus infection in trout fisheries in the UK; such as the presence of an algal bloom, slow rates of stock turnover and

174 ZAFAR IQBAL, RABIA MUMTAZ AND RAMSHA SAJJAD water level dropping < 1 m during summer months. Moreover, high abundance of A. foliaceus shows significant association with high temperature and low water clarity [24]. The infection of two ornamental fishes by Argulus foliaceus observed in the present study may be associated with accidental escape of infected fish into the shipping consignment. Probably the parasites escaped in water containing the fish and remained unnoticed by the consignment dispatchers. Furthermore, inspection of fishes for transport to other places does not normally include conscientious efforts to locate fish lice. The adult fish lice may be obvious on fish being inspected, but intermediate stages may be passed over. Even, the basic health management practices might be easily over looked due to dearth of trained personal, which may cause the shifting of pathogens along with their hosts. This is the first report of argulosis in ornamental fishes imported to Lahore, Pakistan and is a typical example of introduction of potentially serious pathogenic fish parasites from one country to another with their host. This is an alarming situation for the local fish species, aquatic ecosystem and for fish health management experts. Strict regulations on import of healthy and disease free fish are required to be implemented at fish landing sites in the country. This will help to save the emerging ornamental fish trade industry in Pakistan. Acknowledgements: We acknowledge the financial support provided by University of the Punjab, Lahore under Research Project Grant to faculty members for the year 2010-11. Thanks to Dr. Ozturk of Department of Biology, Afyon Kocatepe University, Turkey for his valuable comments on the manuscript. REFERENCES [1]Al-dulaimi, F. H. A., 2010. Infection with fish louse Argulus foliaceus L. in a goldfish (Carassius auratus) at Earthen Ponds and Aquarium Fish in Babylon Province, Iraq. J. Babylon Univ. / Pure and Appl. Sci., 2 (18): 468-673. [2] Anev, N. 2006. New Tendencies for Therapy against parasitic crustacean in fish farms. Bulg. J. Agri. Sci., 12(2): 301-309. [3]Bauer, O. N., V. Musselius and YU. A. Stsrelvok, (1973). Disease of Pond fishes. Israel Program for Scientific Translation, Keter Press Jerusalem. pp230.

ARGULOSIS, IN SOME ORNAMENTAL FISHES IMPORTED TO LAHORE, PAKISTAN 175 [4] Bykhoskaya-pavlovskaya, I. E., A.V. Gusev, M.N. Dubinina, N. A. Izvumova, T.S. smimova, I.L. Sokolovskava, G.A. Shietin, S.S Shulman and V.M Epstein, 1964. Key to Parasites of freshwater Fish of USSR. Acad. Sci. USSR. Zool. Inst. pp. 239-245. [5]Chandra, K. J., 2004. Fish parasitological studies in Bangladesh: A review. J. Agri. Rural Dev., 4(1&2): 9-18. [6] Faruk, M. A. R., M. J. Alam, M.M.R. Sarkar and M. B. Kabir, 2004. Status of Fish Disease and Health Mangement Practices in Rural freshwater aquaculture of Bangladesh. 2004. Pak. J. Biol. Sci.,7(12): 2092-2098. [7] Gresty, K. A, G. A. Boxshell, and K. Nagasawa, 1993. The fine structure and function of the cephalic appendages of the branchiuran parasite Argulus foliaceus. Thiele Phill. Trans. Sco. London B., 339: 119-135. [8] Heckmann, R. A. 1993. Parasites of salmonid fishes. In:, Stoskopt MK (Ed). Fish Medicine., Philadelphia. pp 408-428. [9] Jaffri, S. I. H. and S.S. Ahmad, 1994. Some Observations on Mortality in Major carps due to fish lice and their chemical control. Pak. J. Zool., 26(3): 274-276. [10] Laster, R. G and F. R. Rouball, 1995. Phylum Arthropoda, In: P.T.K Woo (ed.) Fish Diseases and Disorder. Vol.1: Protozoan and Metazoon infection. CAB Inter, Wellingford, UK. pp 475-598. [11] Mickheev, V. N., A. F. Pasternak, E.T. Valtonen and Y. Lankkinen, Y. 2001. Spatial distribution and hatching of overwintered eggs of a fish ectoparasites, Argulus coregoni (Crustacea: Branchiura). Dis. Aqua.Org., 46: 123-128. [12] Menezes, J., M. A. RAMOS, T.G. Pereria and A. Moreria de silva, 1990. Rainbow trout culture in small lakes as a result of massive parasitosis related to careless introductions. Aquaculture., 89: 123-126. [13] Nandp, N. C and S. R. Das, 1991. Argulosis causing juvenile mortality in some fishes at Kakdwip, West Bengal. Ind. J. Fish., 38(2): 132-133. [14] Ozturk, O.M. 2010. An investigation on Argulus foliaceus infection of rudd, Scardinius erythrophthalmus in lake Manyas, Turkey. Scienti. Res. Essays., 5(23), 3756-3759. [15] Pekmezic, G. Z, B. Yardimi, C.S. Bolukbas, Y. E. Beyhan and S. Umur, S. 2009. Morality due to heavy infestation of Argulus foliaceus (Linnaeus, 1758) (Branchiura) in pond-reared carp, Cyprinus carpio L., 1758 (Pisces). Crustaceana., 84 (5-6): 533-557. [16] Poly, W. J., 2008. Global diversity of fish lice (Crustacea: Branchiura: Argulidae) in Fresh water. Hydrobiologia; 595: 209-212. [17] Pritchard, M. H., Kruse, GOW. 1982. The collection and preservation of animal parasites. Univ. of Nebraska Press, Lincoh. USA.

176 ZAFAR IQBAL, RABIA MUMTAZ AND RAMSHA SAJJAD [18] Richards, R. 1977. Diseases of Aquarium fish-2. Skin Diseases. Vet. Rec; 101:132-135. [19] Roberts, R. J. and C. Sommerville, 1982. Diseases of tilapias in: Pullin RSV, Lowe-McConnell RH (eds.). The Biology and Culture of Tilapias. ICLARM Conference Proceedings, 7. ICRAM, Manila. Philippines, pp247-263. [20] Ruane, N., T. K. MCcarty and P. Peilly, 1995. Antibody response to crustacean ectoparasites in rainbow trout Oncorhynchus mykiss Wellbaun), immuned with Argulus foliaceus L. antigen extract. J. Fish Dis., 18: 529-735 [21] Rushton-mellor, S. K., 1992.Discovery of fish louse Argulus japonicas Thiele (Crustacea: Branchiura) in Britian. Aqua. Fish Manage., 23: 269-271. [22] Shauja, K., 2010. A study on the parasitic fauna of imported ornamental fish, Carassius auratus. MS Thesis. Department of Zoology, University of the Punjab, Lahore. Pakistan. pp 56 [23] Taylor, N.G.H., C. Sommerville, and R. Wootten, 2006. The epidemiology of Argulus spp. (Crustacea: Branchura) infections in Stillwater trout fisheries. J. Fish. Dis. 29:193-200. [24] Taylor, N.G.H, R. Wootten, and C. Sommerville, 2009. The influence of Risk factor on the abundance, egg laying habits and impact of Argulus foliaceus in Stillwater trout fisheries. J. Fish. Dis., 32:509-519. [25] Thilakarantne, I. D. S. I. P., G. Rajapaksha, A. Hewakopara, R. Rajapakse and A. C. M. Faizal, 2003. Parasitic infections in freshwater ornamental fish in Sri Lanka. Dis. Aqua. Orgs; 54: 157-162 [26] Toksen, E., 2006. Argulus foliaceus (Crustacea: Branchiura) infection on Oscar, Astronotu ocellatus Cuvier, 1829) and its treatment. E.U. J. Fish. Aqua. Sci., (1-2): 177-179. [27] Walker, P. D., I.J. Russion, R. Duijf, G. Van der velde and S.E. Wandelaar Bonga, 2011. The off-host survival and viability of a native and non-native fish louse Argulus, (Crustacea: Branchiura). Cur. Zool; 57(6): (uncorrected proof). [28] Walker PD, Harris J, van der Velde G, Wendelaar Bonga SE, 2008b. Differential host utilization by different life history stages of the fish ectoparasite Argulus foliaceus (Crustacea: Branchiura). Folia Parasitologica 55: 141 149.

ARGULOSIS, IN SOME ORNAMENTAL FISHES IMPORTED TO LAHORE, PAKISTAN 177 Fig. 1a black moor with a prominent lesion (2.3 x 2.6cm) at caudal peduncle. Fig. 2. A specimen of Argulus foliaceus from black moor

178 ZAFAR IQBAL, RABIA MUMTAZ AND RAMSHA SAJJAD Table 1. Argulus foliaceus infection in ornamental fishes S. No Fish Mean Total length(cm) Mean weight(g) Fish exam. Fish Infect. Prevalence (%) 1 Black moor 8.15±1.59 9.63± 6.53 24 3 12.50 2 Koi 9.95± 2.25 11.45 ±1.5 22 3 13.63 3 Shubunkin 8.44±1.8 6.11±2.62 20 0 - Table 2. Site of attachment of Argulus foliaceus on two ornamental fishes S. Fish Infect. No. of Mean Site of attachment of parasite No fish parasites intensity skin fins head 1 Black moor 3 14 4.66 11(78.57%) 2(14.28%) 1(7.14%) 2 Koi 3 26 8.66 16(61.53%) 7(26.92%) 3(11.53%) Total 6 40 27 9 4 % 67.5 % 22.5% 10%