Transcriptome analysis of grass carp (Ctenopharyngodon idella) and Holland's spinibarbel (Spinibarbus hollandi) infected with Ichthyophthirius multifiliis.

Ichthyophthirius multifiliis is a protozoan ciliate that causes white spot disease (also known as ichthyophthiriasis) in freshwater fish. Holland's spinibarbel (Spinibarbus hollandi) was less susceptible to white spot disease than grass carp (Ctenopharyngodon Idella). In this study, grass carp and Holland's spinibarbel are infected by I. multifiliis and the amount of infection is 10,000 theronts per fish. All grass carp died within 12 days after infection, and the survival rate of Holland's spinibarbel was more than 80%. In order to study the difference in sensitivity of these two fish species to I. multifiliis, transcriptome analysis was conducted using gill, skin, liver, spleen and head kidney of Holland's spinibarbel and grass carp at 48 h post-infection with I. multifiliis. A total of 489,296,696 clean reads were obtained by sequencing. A total of 105 significantly up-regulated immune-related genes were obtained by Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis in grass carp. Cluster of differentiation 40 (CD40), cluster of differentiation 80 (CD 80), tumor necrosis factor-alpha (TNF-α), toll-like receptor 4 (TLR-4), interleukin 1 beta (IL-1ß) and other inflammatory-related genes in grass carp were enriched in the cytokine-cytokine receptor interaction pathway and toll-like receptor pathway. In Holland's spinibarbel, a total of 46 significantly up-regulated immune-related genes were obtained by GO classification and KEGG pathway enrichment analysis. Immune-related genes, such as Immunoglobin heavy chain (IgH), cathepsin S (CTSS), complement C1q A chain (C1qA), complement component 3 (C3) and complement component (C9) were enriched in phagosome pathway, lysosome pathway and complement and coagulation concatenation pathway. C3 was significantly up-regulated in gill and head kidney. Fluorescence in situ hybridization (FISH) showed that the C3 gene was highly expressed in gill tissue of Holland's spinibarbel infected with I. multifiliis. A small amount of C3 gene was expressed in the gill arch of grass carp after infected with I. multifiliis. In conclusion, the severe inflammatory response in vivo after infecting grass carp with I. multifiliis might be the main cause of the death of grass carp. The extrahepatic expression of the gene of Holland's spinibarbel might play an important role in the immune defense against I. multifiliis.

Local immune responses to two stages of Ichthyophthirius multifiliis in ginbuna crucian carp.

Ichthyophthirius multifiliis is a ciliated protozoan parasite and is known to infect many freshwater teleosts. Characterizing the immune system in epithelial tissues, where the parasites penetrate and settle, is key to understanding host-parasite interactions. This study examined local immune responses in vivo to the infective stage (theront and trophont) of the parasites using intra-fin administration, which has been developed to analyze in vivo immune responses using fish fin. CD8α+ and CD4+ T-cell compositions were increased significantly in the fin cavity injected with theront or trophont antigens. The expression of GATA-3 and T-bet mRNA, which regulate differentiation of helper T-cells, was upregulated significantly in leukocytes from the trophont antigen-injected site. In contrast, the percentages of macrophages and neutrophils, which are innate immunity components, were decreased significantly in the injection sites. These results suggest that I. multifiliis antigens inhibit the migration of macrophages and neutrophils, and T-cells are the first responders to I. multifiliis. Thus, to better understand the interaction of host immunity and I. multifiliis, further studies should focus on exploring the inhibitory factors from I. multifiliis or examining innate functions of teleost T-cells.

Blood feast: Exploring the erythrocyte-feeding behaviour of the myxozoan Sphaerospora molnari.

AIMS: As the most abundant cell population in the blood, erythrocytes represent an attractive source of nutrients and a protective niche to a number of pathogens. Previously, we observed the attachment of the myxozoan parasite Sphaerospora molnari to erythrocytes of its host, common carp (Cyprinus carpio), raising a number of questions about the nature of this interaction. METHODS AND RESULTS: We elucidated the impact of S molnari on the number of erythrocytes in healthy and immunocompromised fish, over a period of 6 weeks. While we observed only a mild decrease in RBC numbers in healthy individuals, we witnessed gradual and finally severe haemolytic anaemia in immunosuppressed fish. Accompanying this overt loss was increased erythropoiesis as represented by an increase of erythroblasts in the blood. In vitro, we demonstrated the uptake of host proteins from CFSE-labelled erythrocytes, ultimately inducing death of host RBCs, likely for nutrient gain of the parasite. Nevertheless, the results do not exclude a possible role of erythrocyte-derived proteins in immune evasion. CONCLUSION: Overall, the obtained data provide first evidence for the previously unknown appetite of myxozoan parasites for host erythrocytes and create an important framework for future investigations into the molecular mechanisms underlining this interaction.

Identification of Intracellular Bacteria in the Ciliate Balantidium ctenopharyngodoni (Ciliophora, Litostomatea).

The ciliate Balantidium ctenopharyngodoni is the most prominent protist in the guts of grass carp, where it mainly inhabits the creamy luminal contents of the hindgut. Ciliates are generally colonized by microorganisms via phagotrophic feeding. In order to study the intracellular bacteria in this ciliate, we have successfully established it in in vitro culture. Herein, we investigated and compared the bacterial community structures of cultured and freshly collected B. ctenopharyngodoni. The results showed that these two groups exhibited different bacterial communities. The most abundant bacterial family in freshly collected samples was Enterobacteriaceae, while in cultured samples it was Fusobacteriaceae. In addition, a key intracellular bacterium, Cetobacterium somerae, was identified in the cytoplasm of cultured ciliates using fluorescence in situ hybridization (FISH). This study shows that ciliates can retain the intracellular bacteria acquired in the natural habitat for quite a long time, but the bacterial community structure of ciliates eventually changes after a long period of cultivation.

Oral delivery of Bacillus subtilis spores expressing Clonorchis sinensis paramyosin protects grass carp from cercaria infection.

Clonorchis sinensis (C. sinensis), an important fishborne zoonotic parasite threatening public health, is of major socioeconomic importance in epidemic areas. Effective strategies are still urgently expected to prevent against C. sinensis infection. In the present study, paramyosin of C. sinensis (CsPmy) was stably and abundantly expressed on the surface of Bacillus subtilis spores. The recombinant spores (B.s-CotC-CsPmy) were incorporated in the basal pellets diet in three different dosages (1 × 105, 1 × 108, 1 × 1011 CFU/g pellets) and orally administrated to grass carp (Ctenopharyngodon idella). The immune responses and intestinal microbiota in the treated grass carp were investigated. Results showed that specific anti-CsPmy IgM levels in sera, skin mucus, bile, and intestinal mucus, as well as mRNA levels of IgM and IgZ in the spleen and head kidney, were significantly increased in B.s-CotC-CsPmy-1011 group. Besides, transcripts levels of IL-8 and TNF-αin the spleen and head kidney were also significantly elevated than the control groups. Moreover, mRNA levels of tight junction proteins in the intestines of B.s-CotC-CsPmy-1011 group increased. Potential pathogenetic bacteria with lower abundance and higher abundances of candidate probiotics and bacteria associated with digestion in 1 × 1011 CFU/g B.s-CotC-CsPmy spores administrated fishes could be detected compared with control group. The amount of metacercaria in per gram fish flesh was statistically decreased in 1 × 1011 CFU/g B.s-CotC-CsPmy spores orally immunized group. Our work demonstrated that B. subtilis spores presenting CsPmy on the surface could be a promising effective, safe, and needle-free candidate vaccine against C. sinensis infection for grass carp.

The role of fish helminth parasites in monitoring metal pollution in aquatic ecosystems: a case study in the world's most productive platinum mining region.

Due to the increasing consumption of platinum (Pt), especially in automobile exhaust catalysts, environmental concentrations of Pt are of emerging concern worldwide. Limited information exists on environmental concentrations, particularly in Pt mining regions, while South Africa is the world's main supplier of Pt. Moreover, other metals are also released as by-products of Pt mining, which might also cause environmental concern. Certain fish parasite taxa have the ability to accumulate metals orders of magnitude higher than their hosts and can be used to reliably detect metals with naturally low abundance. Studies on Pt accumulation in parasite-host systems are limited. Therefore, the aims of the present study were (1) to determine the accumulation of a variety of metals (cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), lead (Pb), platinum (Pt), and zinc (Zn)) in helminth fish parasites compared with their hosts from a reference site and an impoundment impacted by Pt mining activities; (2) to assess whether there is a difference between bioaccumulation of metals in infected and uninfected hosts, as well as between hosts with different infection intensities; and (3) to compare the biomarker responses (acetylcholine esterase activity (AChE), metallothionein content (MT), catalase activity (CAT), reduced glutathione content (GSH), malondialdehyde content (MDA), protein carbonyls induction (PC), superoxide dismutase activity (SOD), and cellular energy allocation (CEA)) between infected and uninfected hosts. The cestode Atractolytocestus huronensis accumulated significantly higher concentrations of Cr, Ni, and Pt than their host Cyprinus carpio, while the nematode Contracaecum sp. accumulated significantly higher concentrations of Pt and Zn than their host Clarias gariepinus. Infected fish showed lower metal concentrations compared to uninfected fish, while the parasites had no significant effects on their hosts' biomarker responses. The parasites demonstrated the bioavailability of metals derived from Pt mining activities and their ability to resist its toxic effects. Thus, these parasites are promising sensitive accumulation indicators for Cr, Ni, Pb, and Pt contaminations from Pt mining activities.

High diversity of metazoan parasites in carp gudgeons (Eleotridae: Hypseleotris spp.) from Eastern Australia.

Knowledge of the parasite fauna of Australian freshwater fish is fragmentary and incomplete. An understanding of fish hosts and their associated parasites is vital for the successful management of aquatic ecosystems. In this study, we surveyed the parasite fauna of carp gudgeons (Hypseleotris spp.), a complex of species of Australian freshwater fishes, using morphology and molecular data for the 18S and 28S ribosomal RNA genes. We examined 137 individuals of three different taxa in the carp gudgeon species complex and found 16 parasitic taxa of the Digenea, Cestoda, Nematoda and Arthropoda (five adults and 11 larvae). Eleven parasites are reported for the first time from the carp gudgeons (Pseudodactylogyrus sp., Gyrodactylus sp., Clinostomum sp., Paradilepis patriciae, P. cf. kempi, two unidentified species of Paradilepis, Dendrouterina sp., Parvitaenia sp., two lineages of Cyclophyllidea gen. sp., Procamallanus sp., larvae of a spirurine nematode and Lernaea sp.), in addition to Apatemon cf. hypseleotris Negm-Eldin & Davies, 2001 and the invasive tapeworm Schyzocotyle acheilognathi (Yamaguti, 1934), which were previously reported from these fish hosts. Parasite species richness was double in Lake's and Midgley's carp gudgeons relative to western carp gudgeon. These findings highlight the key role of carp gudgeons as intermediate hosts for multiple parasites with complex life cycles using native birds as definitive hosts and the usefulness of DNA data for the identification of parasite larvae.

Digenean Holostephanus (Trematoda: Digenea: Cyathocotylidae) metacercariae in common carp (Cyprinus carpio Linnaeus, 1758) muscle: zoonotic potential and sensitivity to physico-chemical treatments.

Metacercariae of various species within the genus Holostephanus Szidat, 1936 (Trematoda: Digenea: Cyathocotylidae) occur in muscles of both farmed and wild fish, including common carp (Cyprinus carpio Linnaeus, 1758). The life cycle includes a snail as first intermediate host, fish as second intermediate host and birds or mammals as final hosts. We studied the zoonotic potential and the viability of Holostephanus metacercariae from common carp following exposure to various physical and chemical treatments. Muscle tissue samples of common carp specimens from a fish farm in the north-eastern part of Hungary were examined and metacercariae recovered. The zoonotic potential was evaluated experimentally by using small mammals as models (albino mice, n = 2; and Syrian hamsters, n = 4) infected per os with Holostephanus cysts. Parallelly, Metagonimus metacercariae were used as positive controls. We could not confirm the zoonotic potential of Holostephanus metacercariae as they did not survive in the mammalian intestine whereas Metagonimus metacercariae developed to the adult stage. We assessed the viability of metacercariae isolated from common carp specimens during exposure to different physical treatments (temperatures of -18°C, +20°C, +40°C and +60°C) and chemical agents (5% and 10% acetic acid and 10% sodium chloride (NaCl)). Metacercariae lost viability by freezing at -18°C (2 h), heating at 60°C (20 min), incubation in 5% and 10% acetic acid (5 min) and 10% NaCl (2 h). These methods served as models to investigate the effectiveness of food preparation techniques (such as cold and hot smoking, freezing, salting and pickling) on the survival of metacercariae.

Differential survival of 3rd stage larvae of Contracaecum rudolphii type B infecting common bream (Abramis brama) and common carp (Cyprinus carpio).

The main fish host reaction to an infection with third stage anisakid nematode larvae is a response in which host immune cells (macrophages, granulocytes, lymphocytes) in affected internal organs initially are attracted to the parasite whereafter fibroblasts may enclose the parasite forming granuloma. Generally, the reaction is non-lethal to the parasite which may survive for years in the fish host retaining infectivity to the final host. This may also apply for the anisakid nematode Contracaecum rudolphii (having the adult stage in cormorants, using copepods as first intermediate/paratenic host and zooplankton feeding fish as paratenic hosts). The present study has shown that most Contracaecum rudolphii larvae survive in bream (Abramis brama) (from Lake Balaton, Hungary) whereas the majority of the nematode larvae die in Cyprinus carpio (from Lake Hévíz, directly connected to Lake Balaton). Both cyprinid host species interacted with the nematode larvae through establishing a marked cellular encapsulation around them but with different effects. The differential survival in common carp and bream may theoretically be explained by ecological factors, such as the environmental temperature which either directly or indirectly affect the development of nematode larvae, and/or intrinsic host factors, such as differential immune responses and host genetics.

First ultrastructural and cytochemical data on the spermatozoon and its differentiation in progenetic and adult Archigetes sieboldi Leuckart, 1878 (Cestoda, Caryophyllidea, Caryophyllaeidae).

Spermiogenesis in progenetic and adult stages of Archigetes sieboldi Leuckart, 1878, a tapeworm parasitic in oligochaetes and fish respectively, has been examined using transmission electron microscopy and cytochemical staining for glycogen. General pattern of spermiogenesis is essentially like that of other caryophyllideans, i.e., apical dense material in the zone of differentiation in the early stages of spermiogenesis, rotation of free flagellum and a flagellar bud, and proximo-distal fusion. Interestingly, rotation of a free flagellum and flagellar bud to the median cytoplasmic process (MCP) has been observed unconventionally at > 90° only in progenetic stages. Typical striated roots associated with the centrioles occur rarely in A. sieboldi, and only in form of faint structures in advanced stages of spermiogenesis. In contrast to most caryophyllideans studied to date, penetration of the nucleus into the spermatid body has started before the fusion of the free flagellum with the MCP. This feature has been reported rarely but exclusively in the family Caryophyllaeidae. The unipartite mature spermatozoon of A. sieboldi is composed of one axoneme of the 9 + '1' trepaxonematan pattern with its centriole, parallel nucleus, and parallel cortical microtubules which are situated in a moderately electron-dense cytoplasm with glycogen particles. An unusual arrangement of cortical microtubules in the two parallel rows in region I of the spermatozoon is described here for the first time in the Caryophyllidea. Ultrastructural data on spermiogenesis and the spermatozoon in A. sieboldi from tubuficids and carp are compared and discussed with those in other caryophyllideans and/or Neodermata.

Morphological and molecular characterization of a new species Myxobolus gutturocola n. sp. (Myxozoa: Myxobolidae) from the throat of Hypophthalmichthys molitrix in China.

Myxobolus gutturocola n. sp. was isolated from the throat of silver carp, Hypophthalmichthys molitrix, in Chongqing, China. Myxospore valves are unsymmetrical and smooth. Mature spores are ellipsoidal in frontal view, measuring 12.5 ± 0.2 µm (n = 25) in length, 8.4 ± 0.2 µm (n = 25) in width and 7.1 ± 0.2 µm (n = 25) in thickness. Each spore has two pyriform and unequal sizes polar capsules, the large one with 5.7 ± 0.2 µm in length × 3.6 ± 0.2 µm in width and the small one with 4.6 ± 0.2 µm in length × 2.6 ± 0.1 µm in width. Polar filaments are coiled seven or eight turns in the large polar capsule and four or five turns in the small polar capsule. The coils are arranged almost perpendicularly to the longitudinal axis of the polar capsule. Morphological analysis revealed that M. gutturocola n. sp. is distinct from related species of Myxobolus Bütschli, 1882. Molecular analysis has demonstrated that its SSU rDNA sequences do not match with any available sequences in GenBank. Phylogenetic analysis of the SSU rDNA sequences indicated this species clustered in a clade composed exclusively of parasites infecting the fishes of the Leucisini lineage and most closely related to Myxobolus pavlovskii isolated from the gill filaments of silver carp in Hungary.

Morphological and molecular characterization of Myxobolus dibombensis sp. n. (Myxozoa: Myxobolidae), a parasite of the African carp Labeobarbus batesii (Teleostei: Cyprinidae) from Dibombe River, Cameroon.

Myxobolus dibombensis sp. n. (Cnidaria: Myxosporea: Bivalvulida) is described from the fins of the African carp, Labeobarbus batesii, based on morphological and molecular data. Prevalence of infection was 51.9% (67/129). Ovoid to spherical cyst-like plasmodia were found in the intrasegmental region and among the fin rays. No pathological changes were found in the fish host tissue surrounding the cyst-like plasmodia. Mature myxospores were ovoid in frontal view and lenticular in lateral view, with slightly truncated anterior and rounded posterior ends. Myxospores measured 16.8 (15.8-18.0) µm long and 11.4 (10.0-13.0) µm wide. There was a triangular intercapsular appendix measuring 3.8 (2.6-4.5) µm long. Polar capsules were ovoid and slightly unequal in size, occupying approximately one-third of the myxospore length. The larger polar capsule measured 7 (6-8) µm long and 3.6 (3-4) µm wide, while the smaller one measured 5.8 (4.8-7.0) µm long and 3 (2-4) µm wide. The larger polar capsule contained nine to 11 filament coils, whereas the smaller one contained seven to nine coils. SSU rDNA gene sequence of M. dibombensis sp. n. did not match any sequences available in the GenBank. The similarity with available Myxobolus spp. sequences ranged from 65 to 81%. The novel species clustered with M. algonquinensis, which infects the cyprinid Luxilus cornutus from Canada.

Morphological variation in Myxobolus drjagini (Akhmerov, 1954) from silver carp and description of Myxobolus paratypicus n. sp. (Cnidaria: Myxozoa).

There is uncertainty in the identification of Myxobolus drjagini, the causative agent of silver carp twist disease, in the literature. An investigation of fish parasites in Lake Taihu, China, revealed several Myxobolus drjagini-like myxosporeans infecting the subcutaneous tissue of the head skin, the olfactory and oculomotor nerves in the cranial cavity, and the intrafilamental epithelium of the gills of silver carp Hypophthalmichthys molitrix (Valenciennes, 1844). Myxospores from the head skin and the nerves were identified as conspecific to M. drjagini based on morphological and molecular data; although the spores from each of the two organs presented morphological variations. SSU rDNA sequence analysis revealed that the sequence of M. drjagini previously deposited in GenBank (AF085179) was invalid. Myxospores from the gills were identified as Myxobolus paratypicus n. sp. The spores were oval, asymmetric in frontal view, 13.8 (12.9-14.9) µm long, 9.9 (9.2-11.1) µm wide, and 7.0 µm thick. Two pyriform polar capsules were unequal in size (ratio above 4:1) with slightly converging anterior ends, and the posterior end of the large polar capsule extended beyond the middle of the spore. The large polar capsule was 7.5 (6.2-8.2) µm long and 5.0 (4.2-5.6) µm wide; the small polar capsule was 2.7 (2.1-3.6) µm long and 1.4 (1.1-1.9) µm wide. Polar filaments were coiled with 7-8 turns in the large polar capsule. The SSU rDNA sequence of M. paratypicus n. sp. was not identical to that of any myxozoan available in GenBank and showed highest similarity with M. drjagini (96%) and Myxobolus pavlovskii (95%) collected from bighead carp and silver carp, respectively.

18S and 28S rDNA identity and phylogeny of two novel myxosporeans infecting gills of cyprinid carps inhabiting a cold water wetland in northern India.

The sequences of 18S and 28S rDNA have been extensively used as molecular markers to resolve phylogenetic relationships among various myxosporeans infecting fishes. Two novel myxosporeans have been identified during the present study namely, Henneguya latiusii sp. nov. infecting gills of Crossocheilus latius and Myxobolus atkinsoni sp. nov. infecting gills of Labeo rohita inhabiting Ranjit Sagar Wetland, Punjab (India). These myxosporeans formed plasmodia in the gill arch and gill lamellae respectively and infection was moderate (gill plasmodium index, GPI: 2). Myxospores of both the species possessed distinct morphological and morphometric characteristics hence validated as new species. Phylogenetically, H. latiusii sp. nov. was closest to H. doneci infecting gill filaments of Carassius auratus gibelio, with 96% and 74% similarity in 18S and 28S rDNA respectively. Similarly, M. atkinsoni sp. nov. showed close homogeneity of 90% with M. turpisrotundus and 83% with M. nielii infecting gills of Carassius auratus. The phylogenetic analyses of species of Myxobolus and Henneguya showed a tendency to cluster according to the order or family of the host, tissue and geographical location. This study is the first report on LSU gene marker (28S rDNA) standardized on myxozoans infecting fresh water fishes from Indian subcontinent.

Grass carp which survive Dactylogyrus ctenopharyngodonid infection also gain partial immunity against Ichthyophthirius multifiliis.

Dactylogyrus ctenopharyngodonid and Ichthyophthirius multifiliis are 2 important ectoparasites of fish. Both parasites can induce an immune response in fish that leads to a decrease in parasitic infection intensity and the development of resistance against parasitic reinfection. The present study evaluated whether grass carp Ctenopharyngodon idella that survived a D. ctenopharyngodonid infection could develop immunity against infection by D. ctenopharyngodonid and I. multifiliis. The results demonstrated that when grass carp were infected with D. ctenopharyngodonid, the number of red blood cells and the percentages of thrombocytes, monocytes, and neutrophils in the white blood cells increased significantly in the early stage of infection. The percentage of lymphocytes increased over time following parasitic infection. The mean infection intensity of D. ctenopharyngodonid decreased to 0 on Day 28. The activities of serum acid phosphatase, alkaline phosphatase, lysozyme, and superoxide dismutase increased significantly after D. ctenopharyngodonid infection. In addition, the grass carp that survived a previous D. ctenopharyngodonid infection could completely resist D. ctenopharyngodonid reinfection and partially resist I. multifiliis infection.

First record of the invasive tapeworm, Schyzocotyle acheilognathi (Pseudophyllidea; Bothriocephalidae) in native freshwater fish, Brazil.

Schyzocotyle acheilognathi is a tapeworm cestode commonly found in native freshwater fishes from Asia. This cestode has low host specificity and for that reason it has been registered parasitizing more than 200 cultured and wild fish species, besides amphibians, reptiles and birds from different regions of the world. With a high pathogenic potential, S. acheilognathi may cause mortalities in highly infected fish. In South America, Schyzocotyle was reported in Cyprinus carpio from a Brazilian fish farm at the municipality of Cornélio Procópio, northern Paraná State and from the natural environment in C. carpio from Neuquen River, Patagonia, Argentina. So far, there has been no report of this parasite in South American native fishes. Herein we report the first occurrence of the invasive tapeworm S. acheilognathi in Rineloricaria pentamaculata (Siluriformes, Loricariidae), a native armored freshwater catfish from southern Brazil.

Sequencing, characterization and phylogenomics of the complete mitochondrial genome of Dactylogyrus lamellatus (Monogenea: Dactylogyridae).

Despite the worldwide distribution and pathogenicity of monogenean parasites belonging to the largest helminth genus, Dactylogyrus, there are no complete Dactylogyrinae (subfamily) mitogenomes published to date. In order to fill this knowledge gap, we have sequenced and characterized the complete mitogenome of Dactylogyrus lamellatus, a common parasite on the gills of grass carp (Ctenopharyngodon idella). The circular mitogenome is 15,187 bp in size, containing the standard 22 tRNA genes, 2 rRNA genes, 12 protein-encoding genes and a long non-coding region (NCR). There are two highly repetitive regions in the NCR. We have used concatenated nucleotide sequences of all 36 genes to perform the phylogenetic analysis using Bayesian inference and maximum likelihood approaches. As expected, the two dactylogyrids, D. lamellatus (Dactylogyrinae) and Tetrancistrum nebulosi (Ancyrocephalinae), were closely related to each other. These two formed a sister group with Capsalidae, and this cluster finally formed a further sister group with Gyrodactylidae. Phylogenetic affinity between Dactylogyrinae and Ancyrocephalinae was further confirmed by the similarity in their gene arrangement. The sequencing of the first Dactylogyrinae, along with a more suitable selection of outgroups, has enabled us to infer a much better phylogenetic resolution than recent mitogenomic studies. However, as many lineages of the class Monogenea remain underrepresented or not represented at all, a much larger number of mitogenome sequences will have to be available in order to infer the evolutionary relationships among the monogeneans fully, and with certainty.

Update on the distribution of the co-invasive Schyzocotyle acheilognathi (= Bothriocephalus acheilognathi), the Asian fish tapeworm, in freshwater fishes of Mexico.

The Asian fish tapeworm, Schyzocotyle acheilognathi (syn. Bothriocephalus acheilognathi) represents a threat to freshwater fish, mainly cyprinids, across the globe. This tapeworm possesses an extraordinary ability to adapt to different environmental conditions and, because of that, from its natural geographical origin in mainland Asia, it has colonized every continent except Antarctica. It is thought that this pathogenic tapeworm was first co-introduced into Mexico in 1965 from China, with the grass carp Ctenopharyngodon idella, although the first formal record of its presence was published in 1981. Over the past 35 years, the Asian fish tapeworm has invaded about 22% of the freshwater fish in Mexico. Because fish communities in Mexico are characterized by high species richness and levels of endemism, S. acheilognathi is considered as a co-introduced and co-invasive species. In this review, we update the geographic distribution and host spectrum of the Asian fish tapeworm in Mexico. Up until December 2016, the tapeworm had been recorded in 110 freshwater fish species (96 native and 14 introduced), included in 51 genera, 11 families and 4 orders; it was also widely distributed in all types of aquatic environments, and has been found in 214 localities. We present novel data from a survey aimed at establishing the distribution pattern of the tapeworm in native freshwater fishes of two rivers in north-central Mexico, and the genetic variation among individuals of this co-invasive species collected from different host species and localities. We discuss briefly the factors that have determined the remarkable invasive success of this parasite in freshwater systems in Mexico.

Description and genetic characterization of a new Contracaecum larval type (Nematoda: Anisakidae) from Australia.

Nematode parasites belonging to the genus Contracaecum are economically important parasites with zoonotic significance. Adult Contracaecum spp. are found in the stomach of marine mammals or piscivorous birds, and larval stages infect a wide range of invertebrates and fish species. Human infection with Contracaecum larvae has been reported in Australia and other countries after the consumption of infected fish. Although the genus Contracaecum comprises numerous species, thus far only four Contracaecum larval types have been specifically identified, therefore their life cycle and biology are not yet fully understood. In this study, a new Contracaecum larva (type IV), found in the intestinal tissue of carp caught from Coonancoocabil Lagoon, New South Wales, Australia, is described and characterized genetically. It was identified as Contracaecum bancrofti, a unique Australian species reported previously from the Australian pelican in Northern Territory, New South Wales, Victoria and South Australia. This study highlights the role of migratory birds and introduced fish species, such as carp, in distributing zoonotic pathogens not only across the continent but also from marine to freshwater systems. Coonancoocabil Lagoon is located in the Murrumbidgee Valley National Park, a highly managed conservation area with native fish in wetlands located in close proximity to several fish farms and hatcheries. Infection of a highly resilient invasive fish species, such as carp, with a zoonotic parasite of low host specificity, such as Contracaecum larvae, should be alarming for aquaculture and environmental authorities.

First record of myxozoan parasites from fresh water fishes of Jammu and Kashmir and their pathogenecity.

The present study was aimed to isolate myxozoan parasites infecting the native carp Schizothorax esocinus (Heckel) and pond reared carp Labeo rohita (Ham.) collected from fresh water ecosystems of Jammu and Kashmir from July 2013 to June 2014. Various organs such as eyes, gills, scales, fins, intestines, visceral organs and pericardial cavity of as many as 100 live specimens were examined. Results revealed that 14% of the examined L. rohita were infected with Myxobolus rocatlae (Ham.) and 6% of S. esocinus (Heckel) were infected with M. kashmirensis sp. nov. Clinical signs revealed mucous laden gills while the parasitological observation under stereozoom binocular microscope revealed multiple minute plasmodia on the gills. On rupturing these plasmodia with sharp needle 100-500 myxospores were liberated. The identification to species level was done on the basis of morphological and morphometric attributes of the myxospores. This is the first record of myxozoan parasites in fishes from the state of Jammu and Kashmir, India. Histological observations of infected gills revealed plasmodia of two types (LV4 intralamellar vascular type) in M. rocatlae and (LV3 intralamellar vascular type) in M. kasmirensis causing necrosis, hyperplasia, hypertrophy and vacuolization of the epithelial and vascular endothelium.