Ellipsomyxa Ariusi Sp. Nov. (Myxosporea: Ceratomyxidae), a New Myxosporean Infecting the Gallbladder of Threadfin Sea Catfish Arius Arius in India
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Vol. 142: 83–97, 2020 DISEASES OF AQUATIC ORGANISMS Published online November 19 https://doi.org/10.3354/dao03529 Dis Aquat Org Ellipsomyxa ariusi sp. nov. (Myxosporea: Ceratomyxidae), a new myxosporean infecting the gallbladder of threadfin sea catfish Arius arius in India Archana Chandran*, P. U. Zacharia, T. V. Sathianandan, N. K. Sanil Central Marine Fisheries Research Institute, PO Box 1603, Ernakulam North PO, Kerala 682018, India ABSTRACT: The present study describes a new species of myxosporean, Ellipsomyxa ariusi sp. nov., infecting the gallbladder of the threadfin sea catfish Arius arius (Hamilton, 1822). E. ariusi sp. nov. is characterized by bivalvular, ellipsoid or elongate-oval myxospores with smooth spore valves and a straight suture, arranged at an angle to the longitudinal spore axis. Mature myxospores measured 10.1 ± 0.8 µm in length, 6.8 ± 0.5 µm in width and 7.7 ± 0.7 µm in thickness. Polar cap- sules are equal in size and oval to pyriform in shape. They are positioned at an angle to the longi- tudinal myxospore axis and open in opposite directions. Polar capsules measured 2.8 ± 0.3 µm in length and 2.5 ± 0.4 µm in width; polar filaments formed 4−5 coils, and extended to 32.2 ± 2.1 µm in length. Monosporic and disporic plasmodial stages attached to the wall of gallbladder. Molec- ular analysis of the type specimen generated a 1703 bp partial SSU rDNA sequence (MN892546), which was identical to the isolates from 3 other locations. In phylogenetic analyses, genus Ellip- somyxa appeared monophyletic and E. ariusi sp. nov. occupied an independent position in maxi- mum likelihood and Bayesian inference trees with high bootstrap values. The overall prevalence of infection was 54.8% and multiway ANOVA revealed that it varied significantly with location, year, season, sex and size of the fish host. Histopathological changes associated with E. ariusi sp. nov. infection included swelling, vacuolation and detachment of epithelial layer, re duced mucus production and altered consistency and colour of bile. Based on the morphologic, morphometric and molecular differences with known species of Ellipsomyxa, and considering differences in host and geographic locations, the present species is treated as new and the name Ellipsomyxa ariusi sp. nov. is proposed. KEY WORDS: Myxospores · Gallbladder · Molecular phylogeny · Seasonality · Histopathology · Myxozoa Resale or republication not permitted without written consent of the publisher 1. INTRODUCTION 1995, Shpirer et al. 2018). The genus Ellipsomyxa was established with E. gobii Køie, 2003 re covered Myxosporeans are a diverse group of microscopic from Pomatoschistus microps as the type species endoparasites with more than 2596 nominal species (Køie 2003). Members of the genus are characterised (Okamura et al. 2018). Besides invertebrates, they by smooth, thin-walled, hemispherical or elongate infect a wide range of fishes, and rarely amphibians, myxospores with straight or slightly curved central reptiles, birds and even mammals (Feist & Longshaw transverse sutures (Køie 2003, Whipps & Font 2013). 2006, Chang et al. 2015). Though earlier treated as Mature myxospores are bivalvular with 2 equal, protists, close morphological and molecular affinities spherical polar capsules, with their axes parallel to place them in the phylum Cnidaria (Siddall et al. the sutural line and filaments discharging in opposite *Corresponding author: [email protected] © Inter-Research 2020 · www.int-res.com 84 Dis Aquat Org 142: 83–97, 2020 directions. Species of Ellipsomyxa are generally bralis decrease with increasing age and size of their coelozoic, infecting the gallbladder of fishes (Lom & fish hosts. Haaparanta et al. (1994) reported a higher Dyková 2006). Of the 15 nominal species reported so prevalence of Henneguya creplini in the gills of far, E. apogoni Heiniger & Adlard, 2014, E. manilen- younger Perca fluviatilis and suggested that a lack sis Heiniger & Adlard, 2014, E. nigropunctatis of specific immune re sponses may play a role. Schall Heiniger & Adlard, 2014 and E. arothroni Heiniger & et al. (2018), while studying the spatial and temporal Adlard, 2014 infect ornamental fishes; E. fusiformis variability of Myxo bolus inornatus in smallmouth Davis, 1917 infects elasmobranchs; and E. arariensis bass, observed that the prevalence varied more spa- Zatti et al., 2018, E. amazonesis da Silva et al., 2018, tially than temporally. E. plagioscioni Zatti et al., 2020 and E. paraensis Zatti The present study is part of an ongoing project et al., 2020 infect fresh water fishes. screening the myxosporean parasites infecting fish Though morphological characters play an important destined for human consumption along the south- role in the taxonomy of myxosporeans, limited and west cost of India. Though much information is avail- overlapping morphological features observed in many able on myxosporeans infecting freshwater fishes in genera including Myxidium, Zschokkella, Ellipsomyxa India, limited work has been carried out on these and Sigmomyxa complicate species delineation (Lom parasites in marine and brackish water fishes (Kala - & Dyková 2006, Køie & Karlsbakk 2009, Gunter & vati & Nandi 2007). In the present study, Ellipsomyxa Adlard 2010, Fiala et al. 2015, Zatti et al. 2018). A holis- infections were detected in the gallbladder of the tic approach involving molecular analysis, host/ organ threadfin sea catfish A. arius (Hamilton, 1822) col- specificity and geographical location in addition to lected from different locations along the west coast of morphological features should be taken to enable India. A. arius is a commercially important marine accurate species level identification in myxosporeans. catfish inhabiting estuaries, bar mouths and shallow The majority of myxosporeans do not cause serious coastal waters and feeding mostly on lower inverte- disease in fishes, but some of them are capable of brates (Kailola 1999). Though 2 species of Ellipso - causing mortality, leading to economic losses in many myxa have been reported from freshwater catfishes, cultured species (Yokoyama et al. 2012). Pathological no reports previously existed for marine catfishes. changes associated with gallbladder-infecting myxo - This study is also the first report of an Ellipsomyxa sporeans include swelling, vacuolization and slough- species from Indian waters. In addition, it discusses ing of epithelial cells in the gallbladder (Palenzuela the prevalence and intensity of infection and their et al. 1997). Information on the pathological changes relationship with geographic location, year/ season, induced by genus Ellipsomyxa is limited (Køie 2003, size and sex of the fish host. Køie & Karlsbakk 2009, Azevedo et al. 2013, Whipps & Font 2013, Heiniger & Adlard 2014, Thabet et al. 2016, da Silva et al. 2018, Zatti et al. 2018, 2020). This 2. MATERIALS AND METHODS study elucidates the histopathological changes asso- ciated with Ellipsomyxa infection in the gallbladder 2.1. Sample collection and morphological analysis of the threadfin sea catfish Arius arius. In addition to host size and age, season may also In total, 578 Arius arius were collected using influence the prevalence of myxosporean infections; gill nets from Ernakulam (10° 01’ N, 76° 11’ E), however, studies examining the latter relationship Kozhikode (11° 19’ N, 75° 45’ E), Karwar (14° 48’ N, are limited. Álvarez-Pellitero & Sitjà-Bobadilla (1993) 74° 07’ E) and Goa (15° 09’ N, 73° 57’ E) along the re ported that prevalence of Ceratomyxa labracis southwest coast of India (Fig. 1). The fish were col- increases up to age 1 yr and thereafter shows a lected between January 2016 and De cember 2019; decreasing trend in cultured Dicentrarchus labrax. regular monthly sampling was carried out in Erna - While discussing the frequency distribution and kulam while the frequency of sampling was lower host− parasite relationships of Zschokkella leptathe - for the other 3 sites. The collected fish (398 males rinae infecting atherinid fishes, Su & White (1996) and 180 females) ranged from 15.6 to 31.0 cm in observed a higher prevalence in larger fish hosts. length and 80 to 160 g in weight. The fish collected However, they could not observe any clear relation- from Ernakulam were brought live to the laboratory, ship between seasonal variation and prevalence or maintained in tanks and examined within a week of intensity of infection, or between prevalence and sex capture. Fish collected from other sites were pre- of the fish host. Ryce et al. (2004, 2005) observed that served in ice and brought to the laboratory for pathogenicity and prevalence of Myxobolus cere- detailed studies. Chandran et al.: Ellipsomyxa ariusi sp. nov. in Arius arius 85 2.2. Scanning electron microscopy Contents of infected gall bladders were washed 3 times in distilled water to remove bile salt and other debris. Myxospores were then allowed to adhere on poly-L-lysine-coated coverslips and fixed in 2.5% glutaraldehyde buffered with 0.1 M sodium cacody- late (pH 7.5) for 4 h. The coverslips were washed 3 times in cacodylate buffer, post-fixed in 1% osmium tetroxide for 1 h, then washed 3 times in buffer for 10 min each time. The samples were dehydrated using a series of acetone solutions with increasing concentration (30, 50, 70, 90, 95 and 100%), critical point dried using a Hitachi HCP2 Critical Point Dryer (Hitachi), gold coated using a Quorum SC76220 mini sputter coater (Quorum Technologies), and observed under a Tescan VEGA3 scanning electron micro- scope (TESCAN) at 20 kV. Photomicrographs were taken at various magnifications. 2.3. Prevalence and intensity Multiway ANOVA was carried out to reveal the in- fluence of location, year, season, host size and host Fig. 1. Sampling sites for Arius arius in southern India sex on parasite prevalence. Unlike temperate re gions, seasons in the tropics are classified as pre-monsoon (February to May), monsoon (June to September) and Live fish were killed by neural pithing and exam- post-monsoon (October to January). Fish were grouped ined externally using a Nikon SMZ 1000 stereo- into 5 size classes (11−15, 15−19, 19−23, 23−27 and microscope.