Ligophorus belanaki n. sp. and Ligophorus kederai n. sp. are described from Liza subviridis Valenciennes, 1836 and Valamugil buchanani Bleeker, 1854, respectively. Ligophorus kederai n. sp. has fenestrated ventral anchors, while in L. belanaki n. sp. the ventral anchor is not fenestrated. Ligophorus belanaki n. sp. is similar to L. careyensis, one of its coexisting congeners, in the overall shape and size of hard parts, but differs in having a flat median piece in the structure of the AMP (antero-median protuberance of the ventral bar), copulatory organ with non-ornamented initial part and longer vaginal tube, compared to raised median piece in the AMP, ornamented initial part and comparatively shorter vaginal tube in L. careyensis. Ligophorus kederai n. sp. is similar to L. fenestrum, a coexisting congener, in having fenestrated ventral anchors, but differs in having longer points and narrower base. Ligophorus fenestrum, unlike L. kederai n. sp., also possesses fenestrated dorsal anchors. The principal component analysis (PCA) scatterplots indicate that the two new and eight known Ligophorus species from Malaysian mugilids can be differentiated based on the morphometries of their anchors, ventral bars and copulatory organ separately and when combined together. Numerical taxonomy (NT) analyses based on Jaccard's Index of Similarity and neighbour-joining clustering, is used to facilitate comparison of these two new species with the 50 known Ligophorus based on morphological and metric characters. The two new species are different from each other and the other 50 species in the overall shapes and sizes of hard parts, as indicated by the NT analyses.
The cestode fauna of the darkspotted numbfish, Narcine maculata (Shaw) (Torpediniformes: Narcinidae), from Malaysian Bomrneo was examined for the first time. This work resulted in the discovery of a new genus and two new species of Anteroporidae (Lecanicephalidea). Sesquipedalapex comicus gen. n., sp. n. was erected on the basis of the peculiarities of its scolex, in particular its possession of an extremely long apical modification of the scolex proper, which readily distinguishes it from the other genus in the family. The genus is also distinct in its possession of acetabula that are in the form of suckers, rather than bothridiate in form. This species was found to deeply embed its elongate apical structure for much of its length within the intestinal mucosa, provoking a papilliform expansion of the outer wall of the spiral intestine at the site of attachment. The second new species, Anteropora klosmamorphis sp. n., is readily distinguished from its congeners on the basis of testis number and bothridial shape. Both new species are hyperapolytic. The diagnosis of Anteroporidae is amended to accommodate both new taxa. This increases the total number of genera in the family to two, and the total number of species to five.
Both sexes of Brachiella malayensis n. sp. are described on the basis of specimens found in the nostrils of narrow-barred Spanish mackerel Scomberomorus commerson (Lacepède) collected off Besut, Malaysia. The female of this species closely resembles those of B. magna Kabata, 1968 and B. cybii Pillai, Prabha et Balaraman, 1982 but is distinguishable mainly by the body size and the proportions of the cephalosome, posterior processes and caudal rami. While examining the male, we noticed a systematic inconsistency in some lernaeopodid genera. The genus Brachiella Cuvier, 1830, represented by its type-species Brachiella thynni Cuvier, 1830, and two monotypic genera Charopinopsis Yamaguti, 1963 and Eobrachiella Ho et Do, 1984, represented by Charopinopsis quaternia (Wilson, 1935) and Eobrachiella elegans (Richiardi, 1880), respectively, share distinct synapomorphies in the embracing (vs. pinching) elongate male maxilliped and the female trunk with a pair of long, cylindrical ventroposterior processes (in addition to a pair of modified caudal rami), both of which are involved in their unique reproductive strategy. The latter two genera are herewith relegated to junior synonyms of Brachiella.
Haliotrema susanae sp. nov. is described from the gills of the pinecone soldierfish, Myripristis murdjan off Langkawi Island, Malaysia. This species is differentiated from other Haliotrema species especially those from holocentrids in having a male copulatory organ with bract-like extensions at the initial of the copulatory tube, grooved dorsal anchors and ventral anchors with longer shafts. The maximum likelihood (ML) analysis based on partial 28S rDNA sequences of H. susanae sp. nov. and 47 closely related monogeneans showed that H. susanae sp. nov. is recovered within a monophyletic clade consisting of only species from the genus Haliotrema. It is also observed that H. susanae sp. nov. forms a clade with H. cromileptis and H. epinepheli which coincides with a similar grouping by Young based on solely morphological characteristics. The morphological and molecular results validate the identity of H. susanae sp. nov. as belonging to the genus Haliotrema.
Ninety-five specimens of 14 freshwater fish species from small streams in the Kuala Terengganu district and the Lake Kenyir Reservoir, Malaysia, were surveyed for coccidian infections. Six fish species proved to be infected with apicomplexans belonging to the genus Goussia. In all of these fishes Goussia species were found in unsporulated and semisporulated stages. Oöcysts of four species inhabiting the intestinal epithelium became sporulated in tap-water within 24 hours. In two fish species sporulation failed and only unsporulated oöcysts were recorded in the intestine. Three of the intestinal species finishing sporulation proved to be new to science and were described as Goussia malayensis n. sp., G. bettae n. sp. and G. pogonognathi n. sp. from Apocheilus panchax, Betta splendens and Hemirhamphodon pogonognatus, respectively. The fourth species, found in Trichogaster pectoralis, was identified as G. trichogasteri Székely & Molnár, 1992, a species known from aquarium-cultured T. trichopterus.
The Asian seabass is euryhaline, therefore it is interesting to describe the infestation and survival of caligids at varying salinity on the host. In this study, two different brackish water culture systems with monoculture and polyculture practices were investigated for the occurrence of Caligus spp. on Lates calcarifer. Polyculture practices mainly consisted of snapper (Lutjanus spp.), grouper (Epinephelus spp.) and seabass (L. calcarifer), while the monoculture was stocked with only seabass. A total of 777 Caligus spp. specimens were isolated from the sampling in 2009, consisting of three species; Caligus chiastos, Caligus epidemicus and Caligus rotundigenitalis. In 2011, the total specimen was increased to 3110 and two additional species were found; Caligus punctatus and one unknown species (Caligus sp.). A 98.6% of the total examination was represented by C. epidemicus. Constant presence of C. epidemicus was observed throughout the study, regardless the differences in between culturing practices and systems. This species was able to survive within wide salinity range, from 5 to 28 ppt. The other isolated species (C. chiastos, C. punctatus, C. rotundigenitalis and Caligus sp.) were only found infesting in polyculture cages with the salinity ranging from 25 to 28 ppt. Despite accounts for less than 2% of the total specimens, these species may able to produce a challenge for L. calcarifer polyculture farming activity due to their capability for host switching. The present study revealed the potential risk for cross-species transmission in polyculture practices.
The ciliated protozoan parasite Cryptocaryon irritans infecting marine fishes in Taiwan is described. Developmental characteristics and sequences of the ribosomal DNA regions such as part of 18 S, the entire first internal transcribed spacer, and part of 5.8 S of various Taiwan isolates of C. irritans were investigated. A total of 5 isolates was obtained from different fish-host species and localities, the majority from cultured fish species. C. irritans from Taiwan is able to shift its developmental characteristics, i.e. from non-adherent to adherent tomonts, from individualistic to aggregate-forming tomonts, from infection of the gills only to infection of the gills and body. Thus, it is not possible to classify strains of C. irritans on the basis of these parameters. Premature tomonts that developed from dead fishes were able to produce theronts that could infect fish host. Isolates from Pingtung and the USA had identical nucleotide sequences while an isolate from Malaysia was identical to an Israel isolate. Percentage variation among pairs of Taiwan isolates showed a higher degree of variation than isolate sequences listed in GenBank. Sequence analysis revealed highly aberrant isolates in Taiwan, and a phylogenetic tree distinguished a marine and a low-salinity variant. C. irritans from marine fishes in Taiwan, therefore, display some characteristics not previously reported. Since manipulation of salinity in brackishwater ponds and marine cage sites is not feasible, there is a need to develop new strategies for the control and prevention of cryptocaryoniasis.
In February 2013, forty-seven Notched threadfin bream, the Nemipterus peronii, were sampled from the eastern coastal waters of the South China Sea. The concentration of various elements, namely cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), strontium (Sr), manganese (Mn), selenium (Se), Lead (Pb), nickel (Ni), aluminum (Al), arsenic (As), iron (Fe), and Zinc (Zn) were analyzed in the liver, muscle, and kidney organs of the host, as well as in their parasites Hysterothalycium reliquens (nematode) and the Paraphilometroides nemipteri (nematode), using inductively coupled plasma mass spectrometry (ICP-MS). The former group of parasites showed highest accumulation capacity for Cr, Cu, Fe, Mn, Se, Ni, and Zn while the latter group had high accumulation potential of As, Hg, Cd, Al, Pb, and Sr. The divergence in heavy-metal accumulation profiles of both nematodes is linked with the specificity of microhabitats, cuticle morphology, and interspecific competition. The outcome of this study indicates that both parasite models can be used for biomonitoring of metal pollution in marine ecosystems.
The complete life cycle of a pennellid copepod Peniculus minuticaudae Shiino, 1956 is proposed based on the discovery of all post-embryonic stages together with the post-metamorphic adult females infecting the fins of threadsail filefish Stephanolepis cirrhifer (Monacanthidae) cultured in a fish farm at Ehime Prefecture, Japan. The hatching stage was the infective copepodid. The life cycle of P. minuticaudae consists of six stages separated by moults: the copepodid, four chalimi and adult. In this study, the adult males were observed frequently in precopulatory amplexus with various stages of females however, copulation occurs only between adults. Fertilized pre-metamorphic adult females carrying spermatophores may detach from the host and settle again before undergoing massive differential growth into the post-metamorphic adult female. Comparison of the life cycle of P. minuticaudae has been made with three known pennellids: Lernaeocera branchialis (Linnaeus, 1767), Cardiodectes medusaeus (Wilson, 1908) and Lernaeenicus sprattae (Sowerby, 1806). Among the compared species, P. minuticaudae is the first ectoparasitic pennellid to be discovered to complete its life cycle on a single host without any change in infection site preferences between infective copepodid and fertilized pre-metamorphic female.
Cryptocaryoniasis (also known as marine white spot disease) is mediated by Cryptocaryon irritans. This obligate ectoparasitic protozoan infects virtually all marine teleosts, which includes Lates calcarifer, a highly valuable aquaculture species. Little is known about L. calcarifer-C. irritans interactions. This study was undertaken to gain an informative snapshot of the L. calcarifer transcriptomic response over the course of C. irritans infection. An in-house fabricated cDNA microarray slides containing 3872 probes from L. calcarifer liver and spleen cDNA libraries were used as a tool to investigate the response of L. calcarifer to C. irritans infection. Juvenile fish were infected with parasites for four days, and total RNA was extracted from liver tissue, which was harvested daily. We compared the transcriptomes of C. irritans-infected liver to uninfected liver over an infection period of four days; the comparison was used to identify the genes with altered expression levels in response to C. irritans infection. The greatest number of infection-modulated genes was recorded at 2 and 3 days post-infection. These genes were mainly associated with the immune response and were associated in particular with the acute phase response. Acute phase proteins such as hepcidin, C-type lectin and serum amyloid A are among the highly modulated genes. Our results indicate that an induced acute phase response in L. calcarifer toward C. irritans infection is similar to the responses observed in bacterial infections of teleosts. This response demonstrates the importance of first line defenses in teleost innate immune responses against ectoparasite infection.
Cryptocaryonosis is a major problem for mariculture, and the absence of suitable sero-surveillance tools for the detection of cryptocaryonosis makes it difficult to screen Cryptocaryon irritans-infected fish, particularly asymptomatic fish. In this study, we proposed a serum-based assay using selected C. irritans proteins to screen infected and asymptomatic fish. Eight highly expressed genes were chosen from an earlier study on C. irritans expressed sequence tags and ciliate glutamine codons were converted to universal glutamine codons. The chemically synthesized C. irritans genes were then expressed in an Escherichia coli expression host under optimized conditions. Five C. irritans proteins were successfully expressed in E. coli and purified by affinity chromatography. These proteins were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to screen sera from experimentally immunized fish and naturally infected fish. Sera from both categories of fish reacted equally well with the expressed C. irritans recombinant proteins as well as with sonicated theronts. This study demonstrated the utility of producing ciliate recombinant proteins in a heterologous expression host. An ELISA was successfully developed to diagnose infected and asymptomatic fish using the recombinant proteins as antigens.
The approaches of transcriptomic and proteomic have been widely used to study host-pathogen interactions in fish diseases, and this is comparable to the recently emerging application of metabolomic in elucidating disease-resistant mechanisms in fish that gives new insight into potential therapeutic strategies to improve fish health. Metabolomic is defined as the large-scale study of all metabolites within an organism and represents the frontline in the 'omics' approaches, providing direct information on the metabolic responses and perturbations in metabolic pathways. In this review, the current research in infectious fish diseases using metabolomic approach will be summarized. The metabolomic approach in economically important fish infected with viruses, bacteria and nematodes will also be discussed. The potential of the metabolomic approach for management of these infectious diseases as well as the challenges and the limitations of metabolomic in fish disease studies will be explored. Current review highlights the impacts of metabolomic studies in infectious fish diseases, which proposed the potential of new therapeutic strategies to enhance disease resistance in fish.
This is a catalogue and discussion of the known dactylogyridean monogenean genera of siluriform fishes of the Old World. Of a total of 38 nominal genera, only 19 are considered valid. Seventeen of these 19 genera are currently in the Ancyrocephalidae (containing the Ancyrocephalinae and Ancylodiscoidinae), whilst the other two (Neocalceostoma and Neocalceostomoides) are in the Neocalceostomatidae. The 17 genera are Anchylodiscus, Ancylodiscoides, Bagrobdella, Bifurcohaptor, Bychowskyella, Chauhanellus, Cornudiscoides, Hamatopeduncularia, Mizelleus, Paraquadriacanthus, Pseudancylodiscoides, Protoancylodiscoides, Quadriacanthus, Schilbetrema, Schilbetrematoides, Synodontella and Thaparocleidus. Clariotrema Long, 1981 and Neobychowskyella Ma, Wang & Li, 1983 are considered synonyms of Bychowskyella Akhmerov, 1952, Anacornuatus Dubey, Gupta & Agarwal, 1992 is considered a synonym of Quadriacanthus Paperna, 1961, Mizellebychowskia Gupta & Sachdeva, 1990 is considered a synonym of Neocalceostoma Tripathi, 1959 and Hargitrema Tripathi, 1959 is treated as a synonym of Hamatopeduncularia Yamaguti, 1953. It is proposed that the Ancylodiscoidinae be raised to family status within the order Dactylogyridea to accommodate these 17 'ancyrocephalid' genera from siluriforms, together with Malayanodiscoides and Notopterodiscoides from notopterids. A key and the diagnostic characteristics of the 19 recognised dactylogyridean genera from catfishes plus two from notopterids, together with a list of species and synonyms, are included. New combinations made in this work are Thaparocleidus avicularia (Chen, 1987) n. comb., T. calyciflorus (Chen, 1987) n. comb., T. choanovagina (Luo & Lang, 1981) n. comb., T. dissimilis (Chen, 1988) n. comb., T. leiocassis (Reichenbach-Klinke, 1959) n. comb., T. meticulosa (Chen, 1987) n. comb., T. parasoti (Zhao & Ma, 1999) n. comb., T. persculpus (Chen, 1987) n. comb., T. valga (Chen, 1987) n. comb. and T. wulingensis (Yao & Wang, 1997) n. comb. [all from Silurodiscoides] and Bychowskyella glyptothoraci (Ma, Wang & Li, 1983) n. comb. [from Neobychowskyella].
Culturing fishes in marine cages is a rapidly developing area of marine aquaculture. The Asian seabass Lates calcarifer (Bloch) is a fast growing good quality fish that is readily cultured in intensive systems in the South Asian region and in Malaysia in particular. Although several papers have been published to date on viral, bacterial, parasitic and fungal organisms causing diseases in the Asian seabass, the occurrence of a coccidian infection in this species has only recently been recorded. We collected sporulated and unsporulated oöcysts of a new species of Goussia Labbé, 1986, from the mucus covering the epithelium of the intestine of L. calcarifer. This paper provides a description of Goussia kuehae n. sp. Sporulated oöcysts of this species are ellipsoidal, 37-40 μm in length and 28-30 μm in width. The ellipsoidal sporocysts are relatively small, 15.2-17 × 5.7-8 μm, and located loosely in the oöcyst. There are residual bodies both in the oöcysts and the sporocysts. Goussia kuehae n. sp. differs from all known species of Goussia in the large size of the oöcysts and in having two types of oöcyst residuum.
Monogeneans identified as Sinodiplectanotrema malayanum n. sp. were collected from the fish Pennahia anea (Sciaenidae) off the west coast of Peninsular Malaysia. The new species is recognised on the basis of morphometrical differences in the anchors, marginal hooks and eggs and apparent differences in the 28S rDNA sequence data. The new species possesses features (ovary looping the intestinal caecum, body spines, a vagina and haptoral reservoirs) not noted in the original description of the type and only other species of the genus, S. argyrosomus Zhang, 2001, necessitating the re-assignment of the genus to the Diplectanidae Monticelli, 1903, a move which is supported by 28S rDNA evidence. Sinodiplectanotrema is redefined on the basis of the observation of several features not included in the original diagnosis.
Two new and two previously described species of diplectanid monogeneans (Heteroplectanum flabelliforme n. sp., Diplectanum sumpit n. sp., D. jaculator Mizelle & Kritsky, 1969 and D. toxotes Mizelle & Kritsky, 1969) were collected from archerfish Toxotes jaculatrix off the Island of Langkawi, Kedah and off Perak, Malaysia. The reproductive systems and squamodiscs of D. jaculator and D. toxotes are described for the first time. D. sumpit n. sp. differs from D. toxotes and D. jaculator in a having a small curved copulatory tube with a distinct accessory piece, compared to the long, tubular copulatory tube of D. jaculator and the slender tube of D. toxotes. D. sumpit n. sp. also differs from D. toxotes in having a larger ventral bar and larger squamodiscs. H. flabelliforme n. sp. differs from all known Heteroplectanum species in the shape and size of the squamodiscs, the arrangement of the sclerites in the squamodiscs, the extremely large ventral bar and the short, curved, non-spinous copulatory tube.
Sampling of a large number of elasmobranchs from coastal waters off Borneo revealed the presence of five new species of Dollfusiella Campbell & Beveridge, 1994 (Trypanorhyncha: Eutetrarhynchidae), namely D. angustiformis n. sp., D. hemispinosa n. sp., D. spinosa n. sp., D. imparispinis n. sp. and D. parva n. sp. Dollfusiella angustiformis n. sp. is described from the spiral intestines of four species of the dasyatid stingray genus Himantura Müller & Henle from both the Indonesian and Malaysian parts of Borneo. All the other species were obtained from Malaysian Borneo. Dollfusiella hemispinosa n. sp. is described from the spiral intestines of three species of Himantura, whereas D. spinosa n. sp. was obtained from several specimens of Pastinachus solocirostris Last, Manjaji & Yearsley (Dasyatidae) as well as from Taeniura lymma 1 (sensu Naylor et al., 2012) (Dasyatidae), Neotrygon kuhlii 2 (sensu Naylor et al., 2012) (Dasyatidae), and Glaucostegus cf. typus (sensu Naylor et al., 2012) (Rhinobatidae). Dollfusiella imparispinis n. sp. is described from the spiral intestine of a single specimen of Chiloscyllium punctatum Müller & Henle (Hemiscyllidae) from the South China Sea off Sarawak, whereas D. parva n. sp. was obtained from several species of Himantura. Specimens of the five novel taxa possess scoleces covered with enlarged microtriches, a morphological characteristic exhibited by several other congeners. However, the new species differ from all congeners by possessing unique patterns of oncotaxy as well as combinations of additional morphological features. The number of valid species within Dollfusiella is increased to 26. For this reason, a key for the species of Dollfusiella is provided. Furthermore, novel information on hosts and geographic distribution is provided for two previously described species of Dollfusiella, D. michiae (Southwell, 1929) and D. spinulifera (Beveridge & Jones, 2000). The latter species differs slightly from the original description and shows a much higher variability with regard to the lengths of the scolex and muscular bulbs and the number of testes. These variable characters subdivided specimens of D. spinulifera into relatively distinct groups. However, the specimens did not differ in their oncotaxy and are considered to represent a single variable species.
The high prevalence (80-100%) of the marine leech Zeylanicobdella arugamensis (De Silva) on cage-cultured Asian sea bass Lates calcarifer (Bloch) led us to investigate the percentage of juvenile leeches hatched from deposited cocoons, survival of juvenile and adult marine leeches at different salinity and temperature. The results showed that the hatching percentage of juvenile leeches was highest at salinity of 30 ppt (32.5 ± 2.8%) followed by 20 ppt (18.0 ± 4.3%) and 10 ppt (12.1 ± 1.4%), respectively. It was found that the adult and juvenile leeches could live up to an average range of 4-7 days at salinity ranging from 10 to 40 ppt. The juvenile leeches were able to hatch at temperature ranging from 25 to 35 °C but unable to hatch at 40 °C. The survival period of adult and juvenile leeches ranged from 11 to 16 days at 25 °C, which was comparatively longer than 5-13 days and 10 h--5 days at 27-30 °C and 35-40 °C, respectively. The study provided the information on the physical parameters of salinity and temperature which are most optimal for the marine leech Z. arugamensis to propagate.
Empruthotrema stenophallus n. sp. (Monogenea: Monocotylidae) is described from specimens from the nasal tissue of the blue-spotted maskray Dasyatis kuhlii (Muller and Henle, 1841) collected in shallow waters off Pulau Banggi and Pulau Mabul, Sabah, Borneo, Malaysia. This is the first monogenean species to be described from an elasmobranch collected from Sabah. E. stenophallus can be distinguished from the other 6 members of the genus by the morphology of the sclerotized male copulatory organ, which is narrow, short, and distally tapered. E. dasyatidis Whittington and Kearn, 1992, previously documented from the nasal tissue of several of elasmobranch species from Australia, is recorded from 8 host species distributed around Malaysian Borneo. These represent new host and locality records for this monocotylid. The difficulties in identifying species of Empruthotrema and the apparent lack of host specificity by some members of the genus are discussed.
Cryptocaryon irritans causes Cyptocaryonosis or white spot disease in a wide range of marine fish including Lates calcarifer (Asian seabass). However, the immune response of this fish to the parasite is still poorly understood. In this study, quantitative polymerase chain reaction (qPCR) was performed to assess the expression profile of immune-related genes in L. calcarifer infected by C. irritans. A total of 21 immune-related genes encoding various functions in the fish immune system were utilized for the qPCR analysis. The experiment was initiated with the infection of juvenile fish by exposure to theronts from 200 C. irritans cysts, and non-infected juvenile fish were used as controls. Spleen, liver, gills and kidney tissues were harvested at three days post-infection from control and infected fish. In addition, organs were also harvested on day-10 post-infection from fish that had been allowed to recover from day-4 up to day-10 post-infection. L. calcarifer exhibited pathological changes on day-3 post-infection with the characteristic presence of white spots on the entire fish body, excessive mucus production and formation of a flap over the fish eye. High quality total RNA was extracted from all tissues and qPCR was performed. The qPCR analysis on the cohort of 21 immune-related genes of the various organs harvested on day-3 post-infection demonstrated that most genes were induced significantly (p fish that were allowed to recover from the C. irritans infection (10 days post-infection), expression of the immune-related genes was down-regulated to levels similar to the control fish. These results provide insights into the interaction between C. irritans and L. calcarifer and suggest that the innate immune system plays an important role in early defence against parasite infection allowing the fish to eventually recover from the infection.