Archive of SID Case Report DOI: 10.22067/veterinary.v9i1.62662Received: 2017-Feb-17 Accepted aft er revision: 2017-Jun-15 Published online: 2017-Aug 30 Histopathologic report of infestation by Centrocestus formosanus in Iranian grass carp and

a b c b Annahita Rezaie, Zahra Tulaby Dezfuly, Mehrzad Mesbah, Amin Ranjbar

a Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran b Department of Aquatic Health, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran c Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

Keywords Abbreviations Centrocestus formosanus, grass carp, common C. formosanus: Centrocestus formosanus carp, histopathology H&E: Hematoxylin and Eosin Abstract

Flukes cause severe and lethal diseases in various animals comprising fi sh. Both adult and larval stages are found in fi sh. Centrocestiasis is an in- fection of the fi sh gills by heterophyid trematode Centrocestus formosanus. In summer 2014, 35 fi n- gerling grass carp and 30 common carp weight of 6 grams were referred to the veterinary hospital of Shahid Chamran University of Ahvaz. In the wet mount of the skin, mild infection of trichodina was observed. Wet mount of grass carp gills re- vealed large number of parasitic cysts between gill fi laments. Th e cysts were quite clear and contained pear-shaped parasites. In histopathological exam- ination, fi laments of gill were thick and distorted. According to the wet mount and histopathologic results, metacercariae was chracterised to belong to , C. formosanus.

Corresponding author: IRANIAN JOURNAL OF VETERINARY SCIENCE AND TECHNOLOGY Annahita Rezaie Department of Pathobiology, Tel: +98 51 3880 3742 Fax: +98 51 3876 3852 Faculty of Veterinary Medicine, Web: ijvst.um.ac.ir Email: [email protected] Shahid Chamran University of Ahvaz, Ahvaz, Iran

Email: [email protected]

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Introduction Wet mount of the skin and gills were prepared and studied by light microscopy. Centrocestiasis is an infection of the fi sh gills All fi sh samples submitted for histopathology by heterophyid trematode Centrocestus formosa- were fi xed in 10% buff ered formalin. Th e gill tis- nus which has been described in 1924 from adult sues were processed routinely, embedded in paraf- parasites recovered from natural and experimental fi n, sectioned, stained with Hematoxylin and Eo- defi nitive hosts in Taiwan [19]. sin (H & E), and examined using light microscopy C. formosanus is an exotic parasite native to [13]. Asia that has been introduced into warm water In the wet mount of the skin, mild infestation systems around the world. Th is digenean requires of trichodina was observed. Wet mount of grass three separate hosts to complete its complex lifecy- carp gills revealed large number of parasitic cysts cle. Snail, oft en red-rim melania Melanoides tuber- between gill fi laments. Th e parasitic cysts in wet culatus, is the first intermediate host that utilizes mount of common carp gills were low. Infected the gills of various fishes as second intermediate gill filaments appeared shortened, thickened and host [3]. Th e cycle is completed when a fi sh infect- severely damaged. A clear layer of tissue was ob- ed with metacercariae is eaten by a defi nitive host, served surrounding the cyst. Also, fusions of the piscivorous birds or fi sh-eating mammals. In the filaments were observed. digestive tract of defi nitive hosts, the adult trem- Th e cysts were quite clear and contained pear- atode develops [5]. It appears that this parasite shaped parasites with rapid movements (Figure 1). has a low intraspecifi c variability since no marked An X-shaped excretory bladder was seen (Figure morphological and biometrical diff erences among 2A). Also, they had 32 circumoral spines around worms from the diff erent zoogeographical areas the oral sucker which were arranged in two rows were observed [24]. Metacercarial infection in fi sh is with the sub- sequent economic loss. Metacercariae may aff ect growth and survival, or disfi gure fi sh so that they lose their market cost as a food or ornamental product [10]. Metacercariae have been described from brackish and marine waters in some countries in Asia, particularly from South Korea [2, 7, 25]. Mood et al. (2010) reported Centrocestus formosa- nus metacercarial infection in the gills of diff erent ornamental fish species including Dwarf gourami (Colisalalia), Goldfish (Carassius auratus), Red fin shark (Labeo erythrurus) and Arowana (Osteoglos- Figure 1 sum bicirrhosum) from Iran. Also, Shoaibi Omrani Wet mount of grass carp gill infected with metacercariae of et al. (2010) reported the infection with Ascocotyle C. formosanus. Note diff erent cysts which are located in the tenuicollis in the imported platy fi sh in Iran. fi laments (Arrows). Th e aim of this report was to present clinico- (Figure 2B). pathologic description of Heterophyidae metacer- Histopathologically, fi laments of gill were carial Centrocestus formosanus infection in the gill thick, distorted, and also they were multifocal. of cyprinids in Iran. Th is area was composed of proliferated cartilage Case description and sections of parasite in the center (Figure 3A, B). Th ese metacercarial cysts completely disrupt- In summer 2014, 35 fi ngerling grass carp ed the normal gill morphology. According to wet and 30 common carp with an average weight of 6 mount and histopathologic results, metacercariae grams were referred to the veterinary hospital of was characterized to be C. formosanus. Shahid Chamran University of Ahvaz, Depart- ment of Aquatic Animals Health. Th e clear clinical Discussion symptom was mouth breathing. According to the Unlike the majority of the digeneans, C. for- owner’s statements, the fi sh had high mortality.

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Figure 2 Wet mount of grass carp gill. Metacercariae of C. formosanus. A: Note the X-shaped excretory vesicles (black arrow) and oral sucker (red arrow). B: Spines are located in two rows.

mosanus causes morbidity and mortality in many Th ien et al. (2007) found that severity of me- wild and cultured fish [24]. Th e cercariae of C. for- tacercarial infection was signifi cantly correlated mosanus are highly pathogenic to piscine hosts be- with the smaller body weights in common carp, so cause they encyst in the gills and cause respiratory that small fi sh have a higher prevalence of meta- problems, however this trematode is not highly cercarial infection than larger fi sh [29]. Th e fi nger- pathogenic in mammals, they can cause intestinal lings are more susceptible to trematodes infection. pain, diarrhea, and chronic enterocolitis in man, Th is is may be due to their relatively thin skin, or death in experimental animals if heavily infect- lack of previous contact to infection [12], less gill ed [11]. surface area, thus, requiring fewer metacercariae A massive metacercariae infestation of gill to cause greater disruption of respiratory process, with C. formosanus was the cause of farmed Cich- and having slower defense response than larger lid fi sh’s mortalities [23]. Lauckner (1984) showed fi sh [14]. Diff erences in the age-related quantity or that only a metacercariae of digenean trematode structure of biochemical compounds expelled by was suffi cient to kill a fi sh larva. A Centrocestus fi sh might be another reason for higher infection species was the cause of mortality in aff ected com- in small fi sh [6]. mon carp fry in India [17] and so the mortality Identifi cation of the metacercariae was based of fi sh in this report was probably due to parasitic on characteristic features (Shape of cysts, existence infestation. and size of suckers, and fi gure of excretory blad-

Figure 3 Gill of grass carp. A: Note to distortion of gill lamellae due to multiple hyperplastic tissue (C) and parasitic cysts (aster- isks). B: Parasite is obvious in the center of chondrodysplasia (C) (H & E).

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der) [27]. In this case, X-shaped excretory bladder Confl ict of Interest and 32 circumoral spines help to diagnose type of metacercariae which belongs to C. formosanus. Th e authors have no confl ict of interest to de- Th e histopathological changes of the gills were clare. chondroblastic hyperplasia of the primary lamel- lae and fusion of them in the aff ected gill fi laments. References Th ey envelope the cysts composing parasite. Th is histopathological fi nding was in agreement with 1. Blazer VS, Gratzek JB. Cartilage proliferation in re- Mitchell et al. (2000), Mitchell et al. (2002), Blazer sponse to metacercarial infections of fi sh gills. Journal of and Gratzek (1985) and Olson and Pierce, (1997). Comparative Pathology. 1985 1; 95 [2]:273-80. Th e passage of the cercaria into the gill tissue and formation of a metacercariae induces a reactive 2. Chai JY, Lee SH. Food-borne intestinal trematode infec- tions in the Republic of Korea. Parasitology internation- chondroplasia and inflammationin in the gill. Th e al. 2002 30; 51 [2]:129-54. lesions vastly diminish respiratory capacity. Exposing to unfavorable conditions for ex- 3. Fleming BP, Huff man DG, Bonner TH, Brandt TM. ample environmental factors, and stress increase Metacercarial distribution of Centrocestus formosanus the parasite infection [8]. Recently Alves Pinto et among fi sh hosts in the Guadalupe River drainage of al. (2015) reported that corticosteroid therapy in- Texas. Journal of Aquatic Health. 2011 1; 23 creased development and fecundity of C. formosa- [3]:117-24. nu in treated mice in comparison with untreated mice. Th us, immunosuppressed host may spread 4. Fried B, Graczyk TK, Tamang L. Food-borne intestinal trematodiases in humans. Parasitology Research. 2004 1; fluke eggs in the environment and/or show great- 93 [2]:159-70. er severity of the disease caused by heterophyids, and perhaps other intestinal trematodes. Th us, the 5. Gjurčević E, Petrinec Z, Kozarić Z, Kužir S, Gjurčević monitoring, diagnosis and treatment are essential. Kantura V, Vučemilo M, Džaja P. Metacercariae of In conclusion, this report descried clinico- Centrocestus formosanus in goldfi sh [Carassius auratus pathologic characteristics of metacercarial C. for- L.] imported into Croatia. Helminthologia. 2007 1; 44 [4]:214-6. mosanus infection in the gill of cyprinids in Iran. As the cysts remain for years and they are the source of infection for the defi nitive hosts [4], it 6. Haas W. Physiological analysis of cercarial behavior. Th e Journal of parasitology. 1992 1:243-55. might have important impacts on aquaculture. In order to prevent and control the infection, food 7. Joo CY. Infestation of larval trematodes from fresh-wa- safety in fi sh nurseries, are recommended by Phan ter fi sh and brackish-water fi sh in River Hyungsan, et al. (2010), as keeping fi sh fry in cement or com- Kyungpook Province, Korea. Kisaengch’unghak chapchi. posite tanks in fi ltered water [28], and removal of Th e Korean journal of parasitology. 1984; 22 [1]:78-84. the vector snail by using chemical molluscicides such as copper sulphate in fi sh ponds and circula- 8. Kadlec D, Šimková A, Jarkovský J, Gelnar M. Parasite tion systems [26]. communities of freshwater fi sh under fl ood conditions. Parasitology Research. 2003 1; 89 [4]:272-83. Acknowledgements 9. Lauckner G. Impact of trematode parasitism on the Th e authors wish to thank Dr. Seyed Moham- fauna of a North Sea tidal fl at. Helgoländer Meeresun- tersuchungen. 1984 1; 37 [1-4]:185-99. mad Mousavi for his consultation, and Research Council of Shahid Chamran University of Ahvaz 10. Leatherland JF. Fish diseases and disorders. Woo PT, for fi nancial support. editor. CABI; 2006. Author Contributions 11. Lo CT, Lee KM. Infectivity of the cercariae of Centro- cestus formosanus and pumilio [: Conceived and designed the experiments, Heterophyidae] in Cyprinus carpio. ZOOLOGICAL performed the experiments, and wrote the paper: STUDIES-TAIPEI-. 1996 1; 35: 305-9. Z.T.D., M.M, A.Rajabi. Analyzed the data: A.Re- zaei. 12. Lun ZR, Gasser RB, Lai DH, Li AX, Zhu XQ, Yu XB, Fang YY. Clonorchiasis: a key foodborne zoonosis in

Gill infestation by Centrocestus formosanus IJVST 2017; VOLUME 9; NUMBER 1 52

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China. Th e Lancet infectious 19. Nishigori M. On a new trematode 2000; 143 [1]:185-200. diseases. 2005 31; 5 [1]:31-41. Stamnosoma formosanum n. 3 p., and its life history. Taiwan lgakkai 25. Seo BS, Lee SH, Chai JY, Hong SJ. Zasshr. 1924; 234:181-238. 13. Luna LG. Manual of histologic Studies on intestinal trematodes in staining methods of the Armed Korea XIII. Two cases of natural Forces Institute of Pathology. 20. Olson RE, Pierce JR. A trematode human infection by Heterophy- metacercaria causing gill cartilage opsis continua and the status of metacercarial infection in brackish 14. McDonald DL, Bonner TH, proliferation in steelhead trout water fi shes. Kisaengch’unghak Brandt TM, Trevino GH. Size sus- from Oregon. Journal of wildlife chapchi. Th e Korean journal of ceptibility to trematode-induced Diseases. 1997; 33[4]:886-90. parasitology. 1984; 22 [1]:51-60. mortality in the endangered foun- tain darter [Etheostoma fonticola]. 21. Phan, VT., Ersboll, AK., Nguyen, Journal of Freshwater Ecology. KV., Madsen, H. and Dalsgaard, 26. Shoaibi Omrani B, Ebrahimza- 2006 1; 21 [2]:293-9. A. [2010] Farm-level risk factors deh Mousavi HA, Sharifpour I. for fi sh-borne zoonotic trematode Occurrence and histopathology of Ascocotyle tenuicollis metacercar- 15. Mitchell AJ, Goodwin AE, Salmon infection in integrated small-scale ia in gill of platyfi sh [Xiphophorus MJ, Brandt TM. Experimental fi sh farms in northern Vietnam. maculatus] imported to Iran. Ira- infection of an exotic heterophyid PLoS Neglected Tropical Diseases. nian Journal of Fisheries Sciences. trematode, Centrocestus formo- 4, 742. 2010; 9[3]:472-7. sanus, in four aquaculture fi shes. North American Journal of Aqua- 22. Pinto HA, Mati VL, de Melo AL. culture. 2002 1; 64 [1]:55-9. Experimental centrocestiasis: 27. Sukantason KL, Sukontason K, Worm burden, morphology and Kuntalue B, Boonsriwong N, Piangjai S, Chaithong U, Vanit- 16. Mitchell AJ, Salmon MJ, Huff man fecundity of Centrocestus formo- tanakom P. Surface ultrastructure DG, Goodwin AE, Brandt TM. sanus [: Heterophyidae] of excysted metacercariae of Prevalence and pathogenicity of a in dexamethasone immunosup- Haplorchis taichui [Trematoda: heterophyid trematode infecting pressed mice. Parasitology interna- Heterophyidae]. the gills of an endangered fi sh, tional. 2015 31; 64 [5]:236-9. the fountain darter, in two central Texas spring-fed rivers. Journal of 23. Ramadan RA, Sakr SF, El-Gamal 28. Th ien CP, Dalsgaard A, Nhan NT, Aquatic Animal Health. 2000 1; 12 RM. Prevalence and distribution Olsen A, Murrell KD. Prevalence [4]:283-9. of metacercariae of Centrocestus of zoonotic trematode parasites in Sp., Nishigori, 1924, Trematoda: fi sh fry and juveniles in fi sh farms of the Mekong Delta, Vietnam. 17. Mohan CV, Shankar KM, Ra- Heterophyidae on the gills and Aquaculture. 2009 1; 295[1]:1-5. mesh KS. Mortalities of juvenile other organs of Oreochromis common carp, Cyprinus carpio niloticus fi ngerlings. Suez Canal Veterinary Medicine Journal associated with larval trematode 29. Th ien PC, Dalsgaard A, Th anh BN, [Egypt]. 2002. infection-a case study. Journal of Olsen A, Murrell KD. Prevalence Aquaculture in the Tropics. 1999; of fi shborne zoonotic parasites in 14 [2]:137-42. 24. Scholz T, Salgado-Maldonado G. important cultured fi sh species Th e introduction and dispersal of in the Mekong Delta, Vietnam. Parasitology research. 2007 1; 18. Mood SM, Mousavi HE, Mokhayer Centrocestus formosanus [Nishi- 101[5]:1277-84. B, Ahmadi M, Soltani M, Sharif- gori, 1924] [Digenea: Heterophy- pour I. Centrocestus formosanus idae] in Mexico: a review. Th e American Midland Naturalist. metacercarial infection of four ornamental fi sh species import- ed into Iran. Bull. Eur. Ass. Fish Pathol. 2010 1; 30 [4]:146.

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DOI: 10.22067/veterinary.v9i1.62662

ﮔﺰارش ﻫﯿﺴﺘﻮﭘﺎﺗﻮﻟﻮژي آﻟﻮدﮔﯽ ﺗﻮﺳﻂ ﺳﻨﺘﺮوﺳﺴﺘﻮس ﻓﻮرﻣﻮﺳﺎﻧﻮس در ﮐﭙﻮر ﻋﻠﻒﺧﻮار و ﮐﭙﻮر

ﻣﻌﻤﻮﻟﯽ اﯾﺮان

آﻧﺎﻫﯿﺘﺎ رﺿﺎﯾﯽ1*، زﻫﺮا ﻃﻮﻻﺑﯽ دزﻓﻮﻟﯽ2، ﻣﻬﺮزاد ﻣﺼﺒﺎح3، اﻣﯿﻦ رﻧﺠﺒﺮ2

1ﮔﺮوه ﭘﺎﺗﻮﺑﯿﻮﻟﻮژي، داﻧﺸﮑﺪه داﻣﭙﺰﺷﮑﯽ، داﻧﺸﮕﺎه ﺷﻬﯿﺪ ﭼﻤﺮان اﻫﻮاز، اﻫﻮاز، اﯾﺮان

2 داﻧﺸﺠﻮي دﮐﺘﺮاي ﺗﺨﺼﺼﯽ ﺑﻬﺪاﺷﺖ آﺑﺰﯾﺎن، داﻧﺸﮑﺪه داﻣﭙﺰﺷﮑﯽ، داﻧﺸﮕﺎه ﺷﻬﯿﺪ ﭼﻤﺮان اﻫﻮاز، اﻫﻮاز، اﯾﺮان

3 ﮔﺮوه ﻋﻠﻮم درﻣﺎﻧﮕﺎﻫﯽ، داﻧﺸﮑﺪه داﻣﭙﺰﺷﮑﯽ، داﻧﺸﮕﺎه ﺷﻬﯿﺪ ﭼﻤﺮان اﻫﻮاز، اﻫﻮاز، اﯾﺮان

درﯾﺎﻓﺖ ﻣﻘﺎﻟﻪ: 29/11/1395 ﭘﺬﯾﺮش ﻧﻬﺎﯾﯽ: 1396/03/25

ﭼﮑﯿﺪه

ﺗﺮﻣﺎﺗﻮدﻫﺎ ﻣﻨﺠﺮ ﺑﻪ ﺑﯿﻤﺎريﻫﺎي ﺷﺪﯾﺪ و ﮐﺸﻨﺪه در ﺣﯿﻮاﻧﺎت ﻣﺨﺘﻠﻒ از ﺟﻤﻠﻪ ﻣﺎﻫﯿﺎن ﺷﺪه و در ﻫﺮ دو ﻣﺮاﺣﻞ ﺑﺎﻟﻎ و ﻧﻮزادي در

ﻣﺎﻫﯿﺎن ﯾﺎﻓﺖ ﻣﯽﺷﻮﻧﺪ. ﺳﻨﺘﺮوﺳﺴﺘﯿﺎزﯾﺲ ﯾﮏ ﻋﻔﻮﻧﺖ ﻣﺮﺑﻮط ﺑﻪ آﺑﺸﺶ ﺑﻮده ﮐﻪ ﺗﻮﺳﻂ ﺗﺮﻣﺎﺗﻮد ﺳﻨﺘﺮوﺳﺴﺘﻮس ﻓﻮرﻣﻮﺳﺎﻧﻮس ( .C formosanus) اﯾﺠﺎد ﻣﯽﺷﻮد. در ﺗﺎﺑﺴﺘﺎن ﺳﺎل 1393، 35 ﻣﺎﻫﯽ ﮐﭙﻮر ﻋﻠﻒﺧﻮار و 30 ﮐﭙﻮر ﻣﻌﻤﻮﻟﯽ اﻧﮕﺸﺖ ﻗﺪ ﺑﺎ وزن 6 ﮔﺮم ﺑﻪ

ﺑﯿﻤﺎرﺳﺘﺎن داﻣﭙﺰﺷﮑﯽ داﻧﺸﮕﺎه ﺷﻬﯿﺪ ﭼﻤﺮان اﻫﻮاز، ﺑﺨﺶ ﺑﯿﻤﺎريﻫﺎي آﺑﺰﯾﺎن ارﺟﺎع داده ﺷﺪ. در ﮔﺴﺘﺮش ﻣﺮﻃﻮب ﺗﻬﯿﻪ ﺷﺪه از

ﭘﻮﺳﺖ، آﻟﻮدﮔﯽ ﺧﻔﯿﻒ ﺑﺎ ﺗﺮﯾﮑﻮدﯾﻨﺎ ﻣﺸﺎﻫﺪه ﺷﺪ. ﮔﺴﺘﺮش ﻣﺮﻃﻮب آﺑﺸﺶﻫﺎي ﻣﺎﻫﯿﺎن ﮐﭙﻮر ﻋﻠﻒﺧﻮار ﺗﻌﺪاد زﯾﺎدي اﻧﮕﻞ ﮐﯿﺴﺘﯽ

ﺑﯿﻦ رﺷﺘﻪﻫﺎي آﺑﺸﺸﯽ را ﻧﺸﺎن داد. ﮐﯿﺴﺖﻫﺎ ﮐﺎﻣﻼً ﺷﻔﺎف ﺑﻮد و اﻧﮕﻞ ﮔﻼﺑﯽ ﺷﮑﻠﯽ درون ﻫﺮ ﮐﯿﺴﺖ ﻣﺸﺎﻫﺪه ﺷﺪ. ﺑﺮرﺳﯽ

ﻫﯿﺴﺘﻮﭘﺎﺗﻮﻟﻮژي، ﺿﺨﯿﻢ ﺷﺪن، ﺧﻤﯿﺪه ﺷﺪن رﺷﺘﻪﻫﺎي آﺑﺸﺸﯽ و ﻣﻘﺎﻃﻊ اﻧﮕﻞﻫﺎ را ﻧﺸﺎن داد. ﺑﺮ اﺳﺎس ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ از ﮔﺴﺘﺮش

ﻣﺮﻃﻮب و ﻫﯿﺴﺘﻮﭘﺎﺗﻮﻟﻮژي، ﻣﺘﺎﺳﺮﮐﺮ ﻣﺸﺎﻫﺪه ﺷﺪه ﻣﺮﺑﻮط ﺑﻪ ﺧﺎﻧﻮاده ﻫﺘﺮوﻓﯿﺪه و ﺳﻨﺘﺮوﺳﺴﺘﻮس ﻓﻮرﻣﻮﺳﺎﻧﻮس ﺑﻮده ﮐﻪ ﻣﻨﺠﺮ ﺑﻪ

ﻣﺮگ و ﻣﯿﺮ ﺑﺎﻻي ﻧﺎﺷﯽ از آﻟﻮدﮔﯽ اﻧﮕﻠﯽ ﺷﺪه ﺑﻮد.

واژﮔﺎن ﮐﻠﯿﺪي: ﺳﻨﺘﺮوﺳﺘﻮس ﻓﻮرﻣﻮﺳﺎﻧﻮس، ﮐﭙﻮر ﻋﻠﻒﺧﻮار، ﮐﭙﻮر ﻣﻌﻤﻮﻟﯽ، ﻫﯿﺴﺘﻮﭘﺎﺗﻮﻟﻮژي

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