59 RESEARCH NOTE

NEMATODE INFESTATION IN THE INDONESIAN SHORTFIN () (: ANGUILLIFORMES: ) HARVESTED FROM ACEH WATERS, INDONESIA

Zainal A. Muchlisin, BS, MS, PhD1*, Bahtiar Lubis, BS, MS1, Agung Setia Batubara, BS, MS1, Irma Dewiyanti, BS, MS1, Muzailin Affan, BS, MS, PhD2 and Mufakkir Sidqi, BS, MS3

1Department of Aquaculture, Faculty of Marine and Fisheries; 2Faculty of Mathematics and Natural Sciences; 3Department of Integrated Coastal Zone Management, Institute of Graduate Studies, Syiah Kuala University, Banda Aceh, Indonesia

ABSTRACT

The study was conducted to determine the nature of infestation of on Indonesian shortfin eel (Anguilla bicolor) harvested from Aceh Province waters, Indonesia. The sampling was conducted in Aceh Singkil, Aceh Jaya and Aceh Besar Districts from August to September 2016 and January 2017. Twenty-two samples from Aceh Singkil, one sample from Aceh Jaya, and 23 samples from Aceh Besar were analyzed for gastrointestinal worms. Two species of worms were found in the gastrointestinal tract of fish samples, namely, Anisakis sp. and Procamallanus sp. Anisakis sp. was found in the stomach of the eel sample from Aceh Singkil, while Procamallanus sp. was found in the intestine of the eel samples from Aceh Jaya. No fish samples from Aceh Besar were infested with .

Key words: Anisakis, Indonesia, nematode, Procamallanus, shortfin eel

Philipp. J. Vet. Med., 55(1): 59-64, 2018

INTRODUCTION

Eels of the genus Anguilla are widely found Unfortunately, several species of are throughout the world (Arai and Wong, 2016). threatened by overfishing and habitat loss According to Miller and Tsukamoto (2004), resulting in decrease of glass eel production there are 19 species of eels worldwide. Thirteen (Pujolar et al., 2007; Silfvergrip, 2009). species of eels are reported in tropical regions Information on the distribution and migration (Watanabe et al., 2009); of these, seven species of tropical shortfin eels A. bicolor bicolor and are found in Indonesian waters (Sugeha et al., A. bicolor pacifica have been reported by Arai 2006). Based on biometric characteristics, two et al. (1999), Sugeha and Suhartati (2008), and species of eels have been reported from Aceh Muchlisin et al. (2016); while genetic studies waters, Indonesia: Anguilla bicolor and A. have been conducted by Tanaka et al. (2014) marmorata (Muchlisin and Siti-Azizah, 2009). and Muchlisin et al. (2017). However, no Of the two species, the Indonesian shortfin eel report on parasites, particulary on nematode A. bicolor is the most abundant and distributed infestation in Indonesian shortfin eelA. bicolor widely in Aceh waters (Muchlisin et al., 2017). from Indonesian waters, is available. Eels are catadromous fishes; they migrate The world eels market is very promising. between inland or coastal growth habitats Japan needs about 130,000 tons of eels per and spawn offshore (Arai and Wong, 2016). year, but only 21,000 tons (16.8%) can be met (KKP, 2011). In addition, the global market requires more than 268,000 tons of eels per *FOR CORRESPONDENCE: year (FAO, 2014). Presently, most of the eels (email: [email protected]) produced come from wild populations, and 60 MUCHLISIN et al. production is seasonally dependent. Therefore, on Hurst (1984), Moravec and Justine (2006) eel production has a very good potential for and Koinari et al. (2013). The prevalence and aquaculture business (Muchlisin, 2013). intensity of infection were calculated based The extensive culture of eels has been on Kabata (1985) as follows: prevalence = initiated in Indonesia, where the larvae (total infected fish/total examined fish) × 100; are collected from the wild (Affandi, 2005); intensity = total parasites/total infected fish. therefore, the eels are very susceptible to diseases and parasites harmful to humans. Tumbo et al. (2011) reported that the giant RESULTS AND DISCUSSION marbled eel A. marmorata from North Sulawesi, Indonesia, has been infested with Trichodina A total of 46 eels were examined for sp., Myxobolus sp. and Vorticella sp., while gastrointestinal nematode infestation. Anguillicola crassus is a common parasite of the Twenty-two samples were obtained from Aceh European eel A. anguilla (Evan and Matthews, Singkil, one sample from Aceh Jaya and 23 1999; Kirk, 2003; Kennedy, 2007; Muñoz et samples from Aceh Besar. The fish samples al., 2015). Grano-Maldonado et al. (2011) also from Aceh Singkil and Aceh Jaya were infected reported that the European eel was infected by nematodes with intensity of 1.74-18.0 and by Gyrodactylus anguillae. However, there is prevalence of 86.4-100 (Table 1). No fish sample no information on the parasites that infect the from Aceh Besar was infested with nematode. Indonesian shortfin eelA. bicolor in the waters Further analysis of fish samples from Aceh of Aceh, Indonesia. Thus, the present study Singkil and Aceh Jaya showed that two species was conducted to examine the gastrointestinal of nematodes were detected, namely, Anisakis nematode infestation in Indonesian shortfin sp. and Procamallanus sp. Anisakis sp. was eel A. bicolor harvested from Aceh waters, found in the stomach while Procamallanus Indonesia. sp. was observed in the intestines (Table 2). Anisakis sp. was found in 84% the fish samples from Aceh Singkil, while Procamallanus sp. MATERIALS AND METHODS infested only one fish sample from Aceh Jaya. Eighteen Procamallanus sp. worms were Sampling was conducted from September reported in the fish sample from Aceh Jaya, 2016 to March 2017 in Singkil peat swamp, Aceh while 33 Anisakis sp. worms were found in 19 Singkil District (2o 16’ 32.55” N; 97o 50’ 39.47” fish samples (average of 1.74 worms per fish). E), the Lamno River, Aceh Jaya District (5o 3’ Procamallanus sp. is brown-orange with a 3.79” N; 95o 20’ 26.26” E) and Tibang Reservoir length of 22.35-26.22 mm, with buccal capsule Aceh, Besar District (5o 36’ 4.46” N; 95o 23’ 3.46” in the mouth, esophagus, nerve ring and E). Parasite identification was conducted at intestine. There are alae at the rearmost parts the Laboratory of Fish Quarantine, Aceh Besar of the posterior (Figs. 1 and 2). The length District. Fish samples were captured using of Anisakis sp. ranges from 10.3-12.02 mm. line hooks; they were kept alive individually in It has a mouth with no lip, with ventriculus plastic bags and transported to the laboratory and intestine and a mucron at the end of the for further analysis. All procedures involving posterior part (Figs. 3 and 4). were conducted in compliance with the Anisakis sp. was found in the stomach, and Syiah Kuala University Research and Ethics Procamallanus sp. was detected in the intestine. Guidelines, Section of Care and Use in This indicates that the gastrointestinal tract Research (Ethics Code No: 958/2015). is the most frequently infested organ by these The fishes were measured for total length worms because this organ is rich in nutrients. and body weight and then anesthetized using According to McClelland (2005) Procamallanus MS2222 overdose. Each fish was dissected sp. is not only found in freshwater but also in abdominally, and the gastrointestinal tract marine fishes, and they are commonly detected was removed, placed in a jar, then perforated in the stomach, intestine and pylorus cecum. with a needle, and the gastrointestinal contents This is the first report on the nematode were placed in a petri dish and observed for infection in Indonesian shortfin eel A. bicolor worms with a stereo microscope (Olympus CX- from Aceh Region, Indonesia. The previous 31, Japan) at 100x magnification (Muchlisin et report by Jabal et al. (2015) shows that the al., 2015). Identification of the worm was based eel Anguilla spp. from Lindu lake, Central NEMATODE INFESTATION IN INDONESIAN SHORTFIN EEL 61

Table 1. Prevalence and intensity of nematodes in Indonesian shortfin eelAnguilla ( bicolor) according to sampling locations. Intensity No. of fish No. of Prevalence Location ∑Parasites (parasite/fish samples infested fish (%) sample) Aceh Singkil 22 19 33 1.74 86.4 Aceh Jaya 1 1 18 18 100 Aceh Besar 23 0 - 0 0 Total 46 20

Table 2. Prevalence and intensity of nematodes in Indonesian shortfin eelAnguilla ( bicolor) according to host organs. Parasite No. of No. of Organ Procamallanus Anisakis infected Parasites Intensity Prevalence sp. sp. fish Operculum ------Gill ------Scales ------Pectoral ------fins Annal fins ------Stomach - 33 19 33 1.74 86.4 Intestine 18 - 1 18 18 100 Total 18 33

Sulawesi, Indonesia, were infected by several Carangidae) in New Caledonia (Moravec et al., species of protozoa, e.g. Myxidium sp., 2016). Besides infesting the fish, camallanids Myxobolus sp., Chilodonella sp., Ceratomyxa also infested amphibians in Chennai, India sp., Balantidium sp., Henneguya sp. and (Jeyathilakan et al., 2013) and Lombok Island, Glugea sp., while Gyrodactylus anguillae and Indonesia (Purwaningsih et al., 2016). Anguillicoloides crassus were the common The Anisakis sp. can be identified based on parasites found in the European eel A. anguilla the presence of mucron, boring tooth, and the (Evan and Matthews, 1999; Grano-Maldonado appearance of the esophagus and ventriculus et al., 2011; Kirk, 2003; Kennedy, 2007; Muñoz (Messina et al., 2016), but the boring tooth was et al., 2015). not observed in this study, probably due to low Procamallanus sp. is a brown nematode quality sample appearance. The first report that has a cuticle layer; it has a buccal capsule on Anisakis sp. infection on marine fishes shaped like a barrel and is not split into two (Rastrelliger kanagurta, Decapterus russelli valves (Kabata, 1985). Infestation of worms and Sardinella sirm) in Indonesia was reported in other waters include Camallanus sp., which by Hutomo et al. (1978). Besides infesting infested Awaous stamineus in Hawaii waters the Indonesian eel samples as shown in this (Gagne et al., 2015), Awaous guamensis and study, Anisakis sp. also infested Rastrelliger Kuhlia marginata in New Caledonia (Moravec sp. in Banten Bay and Pelabuhan Ratu waters and Justine, 2006), Xiphophorus maculatus in (Indaryanto et al., 2014; Indaryanto et al., Sao Paulo State, Brazil (Martins et al., 2007), 2015); largehead hairtail Trichiurus lepturus and four species of Carangoides (Osteichthyes: in Kedonganan waters, Bandung (Semarariana 62 MUCHLISIN et al.

Figs. 1-4: 1. The anterior section of Procamallanus sp. showing the mouth, esophagus and nerve ring; 2. The posterior section of Procamallanus sp.; 3. The head and trunk sections of Anisakis sp.; 4. The posterior section of Anisakis sp. showing a mucron.

et al., 2012); mackerel Euthynnus affinis in characterized by severe abdominal pain (Bucci Lhoknga Bay, Aceh Province (Hidayati et al., et al., 2013). Buchmann and Mehrdana (2016) 2016); and several marine fishes from the stated that the clinical symptoms associated southern coast of Kulon Progo, Yogyakarta with Anisakis sp. infection vary from irritation (Setyobudi et al., 2011). of the esophagus and stomach, vomiting and Anisakis sp. is a common parasite with diarrhea, to severe epigastric and abdominal the marine mammal as its final host, marine pain. Therefore, control measures should be invertebrate as an intermediate host, and fish taken to reduce infections of fish stocks and, as transport host (Koie et al., 2005). Hence, the subsequently, the risk of human infections Indonesian shortfin eel serves as a transport (Buchmann and Mehrdana, 2016). host for Anisakis sp., and this parasite can The present study showed that tropical infest humans who consume the undercooked shortfin eel samples from Aceh waters are infested seafood. Once eaten, it can cause infested with two species of nematodes, an acute disease, anisakiasis, which is Anisakis sp. and Procamallanus sp. NEMATODE INFESTATION IN INDONESIAN SHORTFIN EEL 63

(Euthynnus affinis) di tempat pelelangan ikan lhoknga Aceh Besar. Jurnal Medika ACKNOWLEDGEMENT Veterinaria 10(1): 5-8. Hurst RJ. 1984. Identification and description of This study was made possible through the larval Anisakis simplex and Pseudoterranova decipiens (Anisakidae: Nematoda) from New support of the Ministry of Research, Technology Zealand waters. New Zealand Journal of and Higher Education of the Republic of Marine and Freshwater Research 18(2): 177- Indonesia (Kemenristekdikti RI) through the 186. University Excellent Applied Research Scheme Hutomo M, Burhanuddin and Hadidjaja P. 1978. (PTUPT) year 2017 (Contract Number: 81/ Observations on the incidence and intensity UN11.2/PP/SP3/2017). of infection of nematode larvae (Family Anisakidae) in certain marine fishes of waters around Panggang Island, Seribu Islands. REFERENCES Marine Research in Indonesia 21: 49-60. Indaryanto F, Yusli R and Risa T. 2014. Struktur komunitas cacing parasitik pada ikan kembung Affandi R. 2005. Strategi pemanfaatan sumberdaya (Rastrelliger spp.) di perairan Teluk Banten ikan sidat, Anguilla spp. di Indonesia. Jurnal dan Pelabuhan Ratu. Jurnal Ilmu Pertanian Ikhtiologi Indonesia 5(2): 77-81. Indonesia 19(1): 1-8. Arai T, Limbong D, Otake T and Tsukamoto K. Indaryanto F, Yusli R and Risa T. 2015. Inventerisasi 1999. Metamorphosis and inshore migration of cacing parasitik pada ikan kembung di tropical eels Anguilla spp. In the Indo-Pacific. Perairan Teluk Banten dan Pelabuhan Ratu. Journal of Marine Ecology Progress Series Jurnal Veteriner 16(2): 196-203. 182(1): 283-293. Jabal AR, Cahyaningsih U and Tiuria R. 2015. Arai T and Wong LL. 2016. Validation of the Protozoa Parasitik pada Ikan Sidat (Anguilla occurrence of the tropical eels, Anguilla spp.) Asal Danau Lindu, Sulawesi Tengah. bengalensis bengalensis and A. bicolor bicolor Jurnal Ilmu Pertanian Indonesia 20(2):103- at Langkawi Island in Peninsular Malaysia, 107. Malaysia. Tropical Ecology 57(1): 23-31. Jeyathilakan N, Maman M and Jayathangaraj MG. Bucci C, Gallotta S, Morra I, Fortunato A, Ciacci C 2013. Occurrence of Camallanus trispinosus and Iovino P. 2013. Anisakis, just think about in a captive (Geochelone it in an emergency! International Journal of elegans). Journal of Parasitic Diseases 1(1):1-3. Infectious Diseases 17(11): e1071-e1072. Kabata Z. 1985. Parasites and Diseases of Fish Buchmann K and Mehrdana F. 2016. Effects Cultured in the Tropics. London, UK: Tailor & of anisakid nematodes Anisakis simplex Francis, Ltd. (s.l.), Pseudoterranova decipiens (s.l.) and Kennedy CR. 2007. The pathogenic helminth Contracaecum osculatum (s.l.) on fish and parasites of eels. Journal of Fish Diseases consumer health. Food and Waterborne 30(6): 319-334. Parasitology 4: 13-22. Kirk RS. 2003. The impact of Anguillicola crassus Evan DW and Matthews MA. 1999. Anguillicola on European eels. Fisheries Management and crassus (Nematoda, Dracunculoidea); first Ecology 10(6): 385-394. documented record of this swimbladder KKP. 2011. Panduan budidaya ikan sidat. Pusat parasite of eels in Ireland. Journal of Fish penyuluhan kelautan dan Perikanan, Biology 55(3): 665-668. Kementerian Kelautan dan Perikanan, Gagne RB, Hogan JD, Pracheil BM, Mcintyre PB, Rupublic of Indonesia, Jakarta. Hain EF, Gilliam JF and Blum MJ. 2015. Koie M, Berland B and Burt MDB. 1995. Spread of an introduced parasite across the Development to third stage larvae occurs in the Hawaiian archipelago independent of its eggs of Anisakis simplex and Pseudoterranova introduced host. Freshwater Biology 60(2): decipiens (Nematoda, Ascaridoidea, 311-322. Anisakidae). Canadian Journal of Fisheries Grano-Maldonado MI, Gisbert E, Hirt-Chabbert J, and Aquatic Sciences 52(S1): 134–139. Paladini G, Roque A, Bron JE and Shinn P. Koinari M, Karl S, Elliot A, Ryan U and Lymbery 2011. An infection of Gyrodactylus anguillae AJ. 2013. Identification of Anisakis species Ergens, 1960 (Monogenea) associated with (Nematoda: Anisakidae) in marine fish the mortality of glass eels (Anguilla anguilla hosts from Papua New Guinea. Veterinary L.) on the north-western Mediterranean Sea Parasitology 193: 126-133. board of Spain. Veterinary Parasitology 180(3- Martins ML, Garcia F, Piazza RS and Ghiraldelli 4): 323-331. L. 2007. Camallanus maculatus n. sp. Hidayati N, Mutaqien B, Rusli R,Yudha F, (Nematoda: ) in an ornamental Muhammad H and Hambali H. 2016. fish Xiphophorus maculatus (Osteichthyes: Indentifikasi parasit pada ikan tongkol 64 MUCHLISIN et al.

Poeciliidae) cultivated in São Paulo State, Indonesia, with DNA barcoding gene cox1. Brazil. Arquivo Brasileiro de Medicina F1000 Research 6: 258. Veterinária e Zootecnia 59(5): 1224-1230. Muñoz P, Penalver J, De-Ybanes RR and Garcia McClelland G. 2005. Nematoda in Marine J. 2015. Influence of adult Anguillicoloides Parasitology. United Kingdom: CABI crassus load in European eels swimbladder Publishing. on macrophage response. Fish and Shellfish Messina CM, Pizzo F, Santuli A, Buselic I, Boban Immunology 42(2): 221-224 M, Orhanovic SO and Mladineo I. 2016. Pujolar JM, Maes GE and Volckaert FAM. 2007. Anisakis pegreffii (Nematoda: Anisakidae) Genetic and morphometric heterogeneity products modulate oxidative stress and among recruits of the European eel, Anguilla apoptosis-related biomarkers in human cell anguilla. Journal of Bulletin of Marine Science lines. Parasites and Vectors 9: 607. 81(2): 297-308. Miller MJ and Tsukamoto K. 2004. An Introduction Purwaningsih E, Dewi K and Nugroho HA. 2016. to Leptocephali Biology and Identification. Parasitic nematodes of amphibians from Tokyo: Ocean Reeserch Institute, The Lombok Island, Indonesia with description University of Tokyo. of Camallanus senaruensis sp. nov. and Moravec F and Justine J. 2006. Camallanus cotti Meteterakis lombokensis sp. nov. Journal of (Nematoda: Camallanidae), an introduced Coastal Life Medicine 4(9): 708-713. parasite of fishes in New Caledonia. Folia Semarariana I, Suratma WY and Ida INA. 2012. Parasitologica 53: 287-296. Infeksi larva cacing Anisakis spp. pada ikan Moravec F, Gey D and Justine J. 2016. Nematode layur (Trichiurus lepturus). Indonesia Madicus parasites of four species of Carangoides Ventricus 1(2): 293-304. (Osteichthyes: Carangidae) in New Caledonian Setyobudi E, Soeparno and Helmiati S. 2011. waters, with a description of Philometra dispar Infection of Anisakis sp. larvae in some marine n. sp. (Philometridae). Parasite 23: 40. fishes from the southern coast of Kulon Progo, Muchlisin ZA and Siti Azizah MNS. 2009. Diversity Yogyakarta. Biodiversitas 12(1): 34-37. and distribution of freshwater fishes in Silfvergrip AMC. 2009. CITES identification guide Aceh Waters, Northern Sumatra, Indonesia. to the freshwater eels (Anguillidae) with International Journal of Zoological Research focus on the European eel Anguilla anguilla. 5(2): 62-79. Conservation on International Trade in Muchlisin ZA. 2013. Potency of freshwater fishes Endangered Species. Stockholm, Sweden. in Aceh waters as a basis for aquaculture Sugeha HY, Aoyama J and Tsukamoto K. 2006. development program. Jurnal Iktiologi Downstream migration of tropical angullid Indonesia 13(1): 91-96. silver eels in the Poso Lake, Central Sulawesi Muchlisin ZA, Munazir AM, Fuady Z, Winaruddin Island, Indonesia. Journal Limnotek 23(1): 18- W, Sugianto S, Adlim M, Fadli N and Hendri 26. A. 2014. Prevalence of ectoparasites on Sugeha HY and Suharti SR. 2008. Discrimination mahseer fish Tor( tambra Valenciennes, 1842) and distribution of two tropical short-finned from aquacultureponds and wild population of eels (Anguilla bicolor bicolor and Anguilla Nagan Raya District, Indonesia. Human and bicolor pasifica) in the Indonesian Waters. Veterinary Medicine 6(1): 148-152. Prossiding of the Nagisa Westpac Congress Muchlisin, ZA, Fuadi Z, Munazir AM, Fadli N, 1-14. Winaruddin W, Defira CN and Hendri A. Tanaka C, Shirotori F, Sato M, Ishikawa M, Shinoda 2015. First report on Asian fish tapeworm A, Aoyama J and Yoshinaga T. 2014. Genetic (Bothriocephalus acheilognathi) infection of identification method for two subspecies of the indigenous mahseer (Tor tambra) from Nagan Indonesian short-finned eel, Anguilla bicolor, Raya District, Aceh Province, Indonesia. using an allelic discrimination technique. Bulgarian Journal of Veterinary Medicine Zoological Studies 53(1): 1-7. 18(4): 361-366. Tumbo AR, Longdong NS and Kanoli AT. 2011. Muchlisin ZA, Maulidin M, Muhammadar AA and Identifikasi, tingkat indensi, indek dominasi, Putra DF. 2016. Inshore migration of tropical dan tingkat kesukaan parasit pada sidat glass eels (Anguilla spp.) in Lambeso River, (Anguilla marmorata). Biota 16(1): 114-127. Aceh Jaya District, Aceh Province, Indonesia. Watanabe S, Aoyama J, Nishida M and Tsukamoto Aceh Journal of Animal Science 1(2): 58-61. K. 2009. Evaluation of the population structure Muchlisin ZA, Batubara AS, Fadli N, Muhammadar of Anguilla bicolor bicolor using total number AA, Utami AI, Farhana N and Siti-Azizah of vertebrae and the mt DNA control region. MN. 2017. Assessing the species composition Coastal Marine Science 29(2): 165-169. of tropical eels (Anguillidae) in Aceh waters,