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Description of Goussia Kuehae N. Sp. (Apicomplexa: Eimeriidae) Infecting

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Description of Goussia Kuehae N. Sp. (Apicomplexa: Eimeriidae) Infecting Description of Goussia kuehae n. sp. (Apicomplexa: Eimeriidae) infecting the Asian seabass, Lates calcarifer (Bloch) (Perciformes: Latidae), cultured in Malaysian fish farms Csaba Székely, Muhammad Hafiz Borkhanuddin, Faizah Shaharom, Mohd Shukri Adam Embong & Kálmán Molnár Systematic Parasitology An International Journal ISSN 0165-5752 Volume 86 Number 3 Syst Parasitol (2013) 86:293-299 DOI 10.1007/s11230-013-9448-1 Final Edited Manuscript EDIT 3 Click here to download Manuscript: SYPA-D-13-00070 EDIT 3.doc Description of Goussia kuehae n. sp. (Apicomplexa: Eimeriidae) infecting the 1 Asian seabass, Lates calcarifer (Bloch) (Perciformes: Latidae), cultured in 2 3 Malaysian fish farms 4 5 6 7 8 9 Csaba Székely • Muhammad Hafiz Borkhanuddin • Faizah Shaharom • Mohd Shukri 10 Adam Embong • Kálmán Molnár 11 12 13 14 15 16 Received: 25 July 2013 / Accepted: 16 September 2013 17 18 19 20 C. Székely () • M. H. Borkhanuddin • Kálmán Molnár 21 22 Institute for Veterinary Medical Research, Centre for Agricultural Research, HAS 23 24 e-mail: [email protected] 25 26 27 F. Shaharom • M. S. A. Embong 28 29 Institute of Tropical Agriculture (AQUATROP), University Malaysia Terengganu, Kuala 30 31 Terengganu, Malaysia 32 33 34 35 36 Abstract Culturing fishes in marine cages is a rapidly developing area of marine aquaculture. 37 38 The Asian seabass Lates calcarifer (Bloch) is a fast growing good quality fish that is readily 39 40 cultured in intensive systems in the South Asian region and in Malaysia in particular. Although 41 42 several papers have been published to date on viral, bacterial, parasitic and fungal organisms 43 44 causing diseases in the Asian seabass, the occurrence of a coccidian infection in this species has 45 only recently been recorded. We collected sporulated and unsporulated oöcysts of a new species 46 47 of Goussia Labbé, 1986, from the mucus covering the epithelium of the intestine of L. calcarifer. 48 49 This paper provides a description of Goussia kuehae n. sp. Sporulated oöcysts of this species are 50 51 ellipsoidal, 3740 μm in length and 2830 μm in width. The ellipsoidal sporocysts are relatively 52 53 small, 15.2–17 × 5.78 μm, and located loosely in the oöcyst. There are residual bodies both in 54 55 the oöcysts and the sporocysts. Goussia kuehae n. sp. differs from all known species of Goussia 56 57 in the large size of the oöcysts and in having two types of oöcyst residuum. 58 59 60 61 62 63 64 65 2 1 2 Introduction 3 4 5 6 The Asian seabass, Lates calcarifer (Bloch) (Perciformes: Latidae), is a favourite food fish 7 8 cultured mostly in the South Asian region and Australia in intensive and semi-intensive systems. 9 In Malaysia the culture of this species started in 1976, when the Malaysian government has 10 11 embarked on the production using the wild and imported fry specimens in floating net cages in 12 13 Setiu Lagoon on the east coast of Peninsular Malaysia (Norfatimah et al., 2009). Due to the high 14 15 economic importance of Asian seabass culture, its diseases have been studied intensively. 16 17 Among infectious diseases, viral nervous necrosis and a bacterial infection caused by 18 19 Streptococcus iniae Pier & Madin, 1976 are the most important (Nash et al., 1987; Bromage et 20 al., 1999; Maeno et al., 2004; Ransangan & Manin; 2010). Of the parasites infecting L. 21 22 calcarifer, the ciliophoran parasite Cryptocaryon irritans Brown, 1951 plays the highest 23 24 pathogenic role (Diggles & Lester, 1996) but monogeneans such as Benedenia melleni Meyer & 25 26 Grootaert, 1990 and Laticola spp. are also common causative agents of diseases (Denevey et al., 27 28 2001; Yang et al., 2006). In cage cultures copepods such as Lernanthropus latis Yamaguti, 1954 29 30 and Caligus spp. (Maran et al., 2009; Chu et al., 2012; Muhd-Faizul et al., 2012) are frequent 31 causative agents, but sanguinicolid blood flukes can also cause some latent infections (Herbert & 32 33 Shaharom, 1995; Herbert et al., 1995). 34 35 Recently Gibson-Kueh et al. (2011a, b) described a coccidian infection from the intestinal 36 37 epithelium of L. calcarifer in Vietnam and Australia, caused by an epicellularly developing 38 39 Eimeria sp. These authors studied ethanol-fixed sporulated oöcysts, made histological and 40 41 ultrastructure studies and presented data on the molecular structure of the 18S rDNA of this 42 parasite, but due to the lack of fresh material they failed to describe it as a named new species. 43 44 Eimeria spp. infection in Malaysia was first published by Boulard (1977) who described Eimeria 45 46 quentini Boulard, 1977 from the marine fish Aetobatus narinari (Euphrasen). Some freshwater 47 48 coccidians were mentioned by Molnár et al. (2003) who described three new species and 49 50 recorded Goussia trichogasteri Székely & Molnár, 1992 found by Székely & Molnár (1992) in 51 52 an aquarium cultured gouramy in Europe. 53 In a study on parasites of specimens of L. calcarifer cultured in Malaysian cage cultures 54 55 we also found coccidian oöcysts in the intestine of this species which we were able to examine 56 57 from fresh material. The structure of the oöcysts of this coccidium corresponded to that reported 58 59 earlier by Gibson-Kueh et al. (2011a, b). Sporulated oöcysts of this species bear the typical signs 60 61 62 63 64 65 3 of the genus Goussia Labbé, 1986. Based on the morphology of the oöcysts we hereby describe this species as Goussia kuehae n. sp. 1 2 3 4 5 6 Materials and methods 7 8 9 Both the Institute for Veterinary Medical Research in Hungary and University Malaysia 10 11 Terengganu received official approval for the use of fish specimens for parasitological surveys 12 13 from their National Animal Experiment Committees. Fifteen Asian seabass individuals with 14 15 standard length (SL) 1732 cm were selected from cages and carried to the laboratory alive for 16 17 health inspection in February, 2013. Fish were transported to the laboratory in oxygen-filled 18 19 plastic bags and kept in an aquarium for 1–2 days. Following sedation with clove oil they were 20 21 killed with a cervical cut and subjected to complete parasitological examination. To detect 22 coccidian infections, fresh mucus from the intestinal tract was examined for the presence of 23 24 oöcysts. 25 26 Drawings of some sporulated oöcysts were made and measurements were taken 27 28 immediately after the detection of the oöcysts using a Nikon microscope (model MM-800). For 29 30 further studies and for measuring more oöcysts, fresh oöcysts were photographed with a Nikon 31 32 Microscope Model Eclipse 80i advanced microscope; digitised images were obtained and 33 measurements were taken from digitised photos. All measurements are given in micrometres as 34 35 the range followed by the mean and standard deviation in parentheses. 36 37 38 39 40 41 Results 42 43 44 Of the 15 individuals studied for coccidian infections, two specimens (standard length 19 and 23 45 46 cm) proved to be infected. In the larger individual only non-sporulated oöcysts (Fig. 1) were 47 48 recovered, whereas in the smaller specimen oöcysts in advanced stage of sporulation were found, 49 50 in which the sporozoites in sporocysts have already been formed but a relatively large sporocyst 51 52 residuum covered them (Fig. 2). The sporulated oöcysts of the new species are described below. 53 54 55 56 57 58 59 60 61 62 63 64 65 4 Family Eimeriidae Minchin, 1903 1 2 3 4 5 6 Goussia kuehae n. sp. 7 8 Syn. Eimeria sp. of Gibson-Kueh et al. (2011a) 9 10 11 Type-host: Lates calcarifer (Bloch) (Perciformes: Latidae). 12 13 Type-locality: Brackish water cages at Setiu Wetland, Terengganu State, Malaysia. 14 15 Site: Mucus and epithelium of the anterior part of intestine. 16 17 Prevalence of infection: 13.3 %. 18 19 Type-material: Photo types have been deposited in the parasitological collection of the 20 Hungarian Natural History Museum, Budapest (Coll. No. HNHM-70399). 21 22 Etymology: The nomen triviale derives from the name of the first author, Ms S. Gibson-Kueh of 23 24 the Australian-Vietnamese research team who first studied this parasite. 25 26 27 28 Description (Figs. 13) 29 30 31 32 Non-sporulated oöcysts round, 1416 in diameter or slightly elongate-oval, 1820 1416 (Fig. 33 34 1). Sporulated oöcysts (Figs. 2, 3A) (n = 50) ellipsoidal, 3740 (37.9 ± 1.49) in length and 35 36 2830.3 (29.3 ± 0.97) in width. Wall of the oöcyst thin, smooth, colourless. Micropyle absent. 37 Two or 3 pale rounded oöcyst residuum-like bodies 78.5 (7.8 ± 0.54) in size and some round or 38 39 amorphous compact oöcyst residuum-like bodies 13.5 in diameter present (Fig. 2). Oöcyst 40 41 contains 4 ellipsoidal sporocysts (Fig. 3B), 15.2–17.0 × 5.78.0 (16.2 ± 0.7 × 6.7 ± 0.8), located 42 43 very loosely in the oöcyst, leaving large space among them. In most cases they are arranged in 44 45 different directions. Sporocyst wall thin, single-layered. The two sporocyst valves connected by 46 47 an indistinct, longitudinally running suture. Stieda body or Stieda-body-like structures not seen. 48 49 Sporozoites elongate, banana-shaped, located head to tail in the sporocyst, 1415.7 × 2.32.7 50 51 (14.7 ± 0.71 × 2.6 ± 0.19). Sporocyst residuum rough scattered dots (0.8–1.2) arranged in the 52 centre of the sporocyst, partially obscuring sporozoites; size of sporocyst residuum tends to 53 54 permanently decrease during sporulation.
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