Myxozoan Infections Research Articles

Identification and Expression Profiling of Toll-Like Receptors of Brown Trout (Salmo trutta) during Proliferative Kidney Disease.

Proliferative kidney disease is an emerging disease among salmonids in Europe and North America caused by the myxozoan parasite Tetracapsuloides bryosalmonae. The decline of endemic brown trout (Salmo trutta) in the Alpine streams of Europe is fostered by T. bryosalmonae infection. Toll-like receptors (TLRs) are a family of pattern recognition receptors that acts as sentinels of the immune system against the invading pathogens. However, little is known about the TLRs’ response in salmonids against the myxozoan infection. In the present study, we identified and evaluated TLR1, TLR19, and TLR13-like genes of brown trout using data-mining and phylogenetic analysis. The expression pattern of TLRs was examined in the posterior kidney of brown trout infected with T. bryosalmonae at various time points. Typical Toll/interleukin-1 receptor protein domain was found in all tested TLRs. However, TLR13-like chr2 had a short amino acid sequence with no LRR domain. Phylogenetic analysis illustrated that TLR orthologs are conserved across vertebrates. Similarly, a conserved synteny gene block arrangement was observed in the case of TLR1 and TLR19 across fish species. Interestingly, all tested TLRs showed their maximal relative expression from 6 to 10 weeks post-exposure to the parasite. Our results suggest that these TLRs may play an important role in the innate defense mechanism of brown trout against the invading T. bryosalmonae.

Myxobolus episquamalis infection in farmed flathead grey mullet Mugil cephalus L. and thin-lipped mullet Liza ramada

Egypt is the largest producer of farmed mullets worldwide. The mullet seeds are collected from natural waters at the north and northern east of Egypt. Thus, the occurrence of parasitic diseases is relatively high even after being stocked in aquaculture facilities. The current study was conducted on a fish farm located at the coastal area of the Suez Canal region, Egypt. Farmed flathead grey mullet (Mugil cephalus L.) and thin-lipped grey mullet (Liza ramada) showed pinkish-white granular cysts on their body surfaces that measured approximately 5 mm (4–6 mm) in width and 7 mm (5–9 mm) in length and contained a milky coloured exudate. The proximal portion of the fins and apical part of the scales were the only sites of infection. These cysts are randomly scattered throughout the fish body surface. No mortalities were detected among moribund mullets, but these cysts make the fish unsightly, causing commercial rejection. The overall prevalence of infection was 40%. Giemsa-stained wet smears initially indicated that the spores were of Myxobolus species. Subsequently, the phylogenetic analysis of the 18S rRNA gene confirmed the identification of spores as being those of Myxobolus episquamalis. Furthermore, amprolium-salinomycin mixture, which has an established track record for treating myxozoan infections in multiple fish species, was successfully tested in a large-scale field trial. Moribund mullets displayed significant reduction in myxosporean infection following treatment with amprolium-salinomycin mixture. To the best of our knowledge, this is the first report describing the detection of M. episquamalis in farmed flathead grey mullet (Mugil cephalus L.) and thin-lipped grey mullet (Liza ramada) collected from fish farms in the Suez Canal, Egypt.

First Myxozoan Infection (Cnidaria: Myxosporea) in a Marine Polychaete from North America and Erection of Actinospore Collective Group Saccimyxon

In a survey of marine annelids for myxosporean infection in Charleston Harbor, South Carolina, we collected 3,214 polychaetes from 21 families and found infections in 6 spionid individuals. Based on gross morphology and COI sequencing, all infected spionids were identified as Streblospio benedicti. Infection prevalence was 0.8% (6/734) of that species of spionid, and 0.2% of all 3,214 polychaetes examined. Pansporocysts contained 8 actinospores and developed in the tegument of the annelid host. This is the first myxozoan infection recorded from this polychaete species, second in the family, and the first marine myxozoan found in the Americas. It is the first marine species found to develop in the tegument of its annelid host; a site of development observed only once before, in Ceratonova shasta infections of its freshwater sabellid polychaete host. Mature actinospores were morphologically simple, truncated ellipsoids, lacking processes or ornamentation, 9.0 ± 0.5 μm × 6.0 ± 0.4 μm. Their sack-like shape was similar to 9 of the 12 actinospores described previously from polychaetes; 10/12 had been and ascribed originally to the morphological collective group Tetractinomyxon despite 9 of these having few similarities to the original description of this group. We propose to name the simple, spherical to ellipsoidal spore morphotype Saccimyxon to encompass both our novel actinospore and the 9 other sack-like polychaete actinospore types in the literature. In the present study, 18S rDNA sequencing demonstrated that the myxozoans that infected the 6 spionids were genetically the same species (type sample 1,737 nucleotides, GenBank accession number MH791159) and was not >95% similar to any sequence in GenBank. Phylogenetic analysis showed that the myxozoan species we encountered is basal to the kudoids and thus likely to have a morphologically simple myxospore stage with fewer than 4 valves. However, without a genetic match, the presumptive vertebrate host remains unknown.

Molecular Genetic Studies on Myxobolus cylindricus and Henneguya mystasi (Myxosporea: Myxobolidae) Infecting Two Indian Fish Species, Channa gachua and Mystus vittatus, Respectively.

Myxozoan infections of Indian freshwater fishes are relatively well studied, but their validity is supported with molecular and phylogenetic data only for a few species. The primary objective was to present molecular data for two Myxozoan species, Myxobolus cylindricus and Henneguya gachua collected from Indian freshwater fishes, the dwarf snakehead Channa gachua and the striped dwarf catfish Mystus vittatus, respectively. Various organs of 56 C. gachua and 48 M. vittatus were dissected. Myxozoan plasmodia with mature spores were collected from the gills under a dissecting microscope. Spores obtained from plasmodia from infected hosts were fixed in 80% ethanol in vials and sent for further morphological and molecular examinations to Hungary. The 18S rDNA gene of Myxobolus and Henneguya spp. was amplified using nested polymerase chain reaction (PCR). Phylogenetic analysis was performed using maximum likelihood (ML) and Bayesian inference (BI) methods. Morphological characteristics of M. cylindricus and H. mystasi spores corresponded to the original descriptions made by Sarkar, Mazumdar and Pramanik, 1985 and Haldar, Samal, and Mukhopadhyay, 1997, respectively. Phylogenetic analysis of the 18S rDNA gene revealed that the sequences of M. cylindricus differed from those of most Indian Myxobolus sp., known mostly from cyprinid fishes and formed a subgroup with Myxobolus neurophilus, a parasite of a perciform host, and with Henneguya chaudhuryi, a species belonging to a different genus but described from a closely related channid host. It was also closely related to another Henneguya species, H. lesteri, described from Sillago analis, a coastal fish. Henneguya mystasi had the closest similarity to Henneguya bicaudi, a species described from an Indian cyprinid fish and to Henneguya pellucida reported from a characid fish known from South America. Molecular data received by us gives a solid basis for further identification of these myxozoans, the pathogenicity of which probably plays an economic role at culturing the hosts.

Morphological, histological, and molecular description of Myxobolus ompok n. sp. (Myxosporea: Myxobolidae), a kidney myxozoan from Pabdah catfish Ompok pabda (Hamilton, 1822) (Siluriformes: Siluridae) in India.

In a parasitological survey of freshwater fishes near Meerut, Uttar Pradesh, India, myxozoan infections in Pabdah catfish Ompok pabda Ham. (Siluriformes: Siluridae) were found. Round plasmodia and scattered spores of Myxobolus ompok n. sp. were found in the kidney of the host. Plasmodia measuring 150-200μm were located in the renal interstitium. Spores of Myxobolus ompok n. sp. were elongated pyriform, 13.6-14.4 (14.8 ± 0.42) μm long, 5.6-6.4 (6.5 ± 0.33) μm wide, and 5.2-6.4 (5.9 ± 0.43) μm thick with two equal polar capsules measuring 8.0-8.5 (8.2 ± 0.2) μm in length and 1.5-2.4 (1.8 ± 0.33) in width having six filamental turns. Both the morphology and DNA analysis of the 18S rRNA gene revealed that Myxobolus ompok n. sp. is distinct from previously described species of Myxobolus and shares no significant similarity with any other Myxobolus deposited in the GenBank database. Phylogenetic analysis inferred that this species showed the closest similarity to Myxobolus miyarii (KT001495). This is the first record of any Myxobolus sp. from O. pabda in India.

Morphological and ssrDNA sequence based molecular characterization of a novel Thelohanellus species (Myxosporea: Myxobolidae) infecting the fins of Goldfish, Carassius auratus L. with special reference to its histopathological alteration

A new species of myxozoan, Thelohanellus goldi n. sp. is described using morphological and molecular data, parasitizing the fin filaments from 18 of 25 host specimens (72.5%) of Carassius auratus collected from different ornamental fish farms of India. Mature spore of the new species were oval to spherical in frontal view having rounded posterior ends and tapering anterior end measures 8.7–10.26 (9.50) × 4.10–7.89 (5.84) μm. The single large polar capsule, round to oval in shape but slightly pointed at the anterior end measuring 4.91–7.63 (5.60) × 2.3–3.1 (2.96) μm and located just below the anterior end of the spore. Polar filament only at distal end with 5–6 loose coils. The most differentiating feature from closely related species was carried out by morpho-taxonomic affinities with previously described species which are tremendously supported by molecular taxonomy by partial sequencing of the 18S rDNA gene resulted in a total of 2124 bp fragment of newly obtained small subunit ribosomal RNA gene sequence of the new species which Exhibit 93–95% homogeneity with other closely related species available in GenBank. The BLAST search and high genetic diversity of distance matrix of Myxobolus sp. did not properly match with any available sequences in GenBank and make sister clade with Thelohanellus caudatus and Thelohanellus habibpuri in the Thelohanellus clade including most of Thelohanellus spp. The study of evolutionary history enables us to understand the evolution of modern species and supports some uncertain topologies which are being presented regarding the morphometric analysis. The severity of myxozoan infection has been assessed in this article by observing the histopathological changes of fins of the C. auratus along with the diversity, distribution and taxonomic description of the new Thelohanellus species with their new host and locality records.

Bromodeoxyuridine DNA labelling reveals host and parasite proliferation in a fish-myxozoan model.

Enteromyxum leei is a myxozoan parasite responsible for enteritis in gilthead sea bream (Sparus aurata). The parasite proliferates in the paracellular space of the intestinal epithelium and induces an inflammatory reaction. To assess intestinal cell turnover and parasite proliferation, fish were infected with the parasite by anal intubation; after 17 and 64days, the cell proliferative marker bromodeoxyuridine (BrdU) was administered; and after 24hr, tissue samples were taken for immunohistochemical detection. Parasite exposure induced increased epithelial and immune cell proliferation in all intestinal segments at all time points, even before parasite establishment. This increased turnover was triggered early after intubation and mainly at a local level, as shown by an increased proliferating cell nuclear antigen (pcna) gene expression only at the posterior intestine after 17days (not found in lymphohaematopoietic organs). Incorporation of BrdU in parasite secondary and tertiary daughter cells indicated that parasite endogeny is not by schizogonial division, which uses de novo synthesis pathway of pyrimidines. Altogether, BrdU immunolabelling and pcna gene expression showed the rapid proliferative response of the fish intestines upon a myxozoan infection and how this response is effectively triggered even before the parasite reaches or establishes in the site.

Morphological and molecular description of Myxobolus batalhensis n. sp. (Myxozoa, Myxosporea), a liver and ovary parasite of Salminus hilarii in Brazil.

Plasmodia containing myxospores belonging to the genus Myxobolus Bütschli, 1882 were found in the ovaries and liver of Salminus hilarii. Despite its economic value, this fish host has no previous reports of myxozoan infections. Herein, we describe Myxobolus batalhensis n. sp. using morphological and ultrastructural data, as well as histological and SSU rDNA molecular data. The mature myxospores were elongated, measuring in average 15.2±0.8μm in length, 8.4±0.4μm in width, and 5.1±0.2μm in thickness. Polar capsules were elongated and measured 5.3±0.3μm in length and 2.8±0.3μm in width. Polar filaments had 6-9 coils. Histopathological analysis showed coagulation necrosis associated with cell lysis as a response of the host cell to the parasite in the ovaries. No inflammatory reaction was observed in the liver, although the presence of the plasmodia caused changes in tissue structure. The phylogenetic analysis of South American myxobolid species showed M. batalhensis n. sp. as sister species of Myxobolus aureus. This is the first report of a myxozoan species parasitizing S. hilarii and the first myxozoan species described in the Batalha river.

Myxosporidiosis (Myxozoan Infections) in Warmwater Fish

Discusses myxozoans--metazoan parasites of marine and freshwater invertebrates and fishes. Provides examples of species that infect aquariumand warmwater species; morphology; general life cycle and transmission; methods for diagnosing infections in fish; and advice for treatment, prevention, and control.

The impact of Tetracapsuloides bryosalmonae\xa0and\xa0Myxobolus cerebralis co-infections on pathology in rainbow trout

BackgroundMyxozoan parasites pose emerging health issues for wild and farmed salmonid fish. Rainbow trout (Oncorhynchus mykiss) is a particularly susceptible species to Tetracapsuloides bryosalmonae (Malacosporea), the etiological agent of Proliferative Kidney Disease (PKD), and to Myxobolus cerebralis (Myxosporea), the etiological agent of Whirling Disease (WD). The objective of this study was to investigate the impact of myxozoan co-infections on the pathogenesis of PKD and WD in the rainbow trout.MethodsTwo groups of rainbow trout (96 fish each) were primarily infected with T. bryosalmonae and triactinomyxons of M. cerebralis; after 30 days half of the fish in each group were co-infected with these parasites vice versa and remaining half was continued as single infection. Mortalities and clinical signs were recorded at different time points. Histopathology and immunohistochemistry were performed to assess the extent of each infection and estimate the parasite burden between groups.ResultsFish firstly infected with M. cerebralis and co-infected with T. bryosalmonae exhibited exacerbated pathological changes of both parasitic diseases and elicited a higher mortality rate. A higher kidney swelling index (grade 4) appeared together with more severe cartilage destruction and displacement, when compared to the pathological changes in fish upon single infections with T. bryosalmonae or M. cerebralis. Conversely, fish firstly infected with T. bryosalmonae and co-infected with M. cerebralis also exhibited typical pathological changes of both parasitic diseases, but with a lower mortality rate, similar as caused by the single T. bryosalmonae or M. cerebralis infection. WD clinical signs were milder, without skeletal deformities, while kidney swelling index was similar to single infection with T. bryosalmonae (grade 2 to 3).ConclusionsIn this study, a co-infection with myxozoan parasites was for the first time successfully achieved in the laboratory under controlled conditions. The impact of co-infections in concurrent myxozoan infections mainly depends on the primary pathogen infecting the host, which could alter the outcomes of the secondary pathogen infection. The primary M. cerebralis infection followed by T. bryosalmonae had a much more serious impact and elicited a synergistic interaction. Contrasting results were instead seen in rainbow trout primarily infected with T. bryosalmonae and then co-infected with M. cerebralis.

Myxobolus sp. and Henneguya sp. (Cnidaria: Myxobolidae) natural co-infection in the kidney of Piaractus mesopotamicus (Characiformes: Serrasalmidae).

This study evaluated the myxozoan infection and histopathology of the kidney of freshwater fish Piaractus mesopotamicus from intensive fish farming in Brazil. A total of 55 fish were examined for myxozoan infection. Infected organs were processed by usual histology and stained with hematoxylin-eosin (H&E) and Ziehl-Neelsen (ZN). From the total of 55 fish analyzed, 47 (85.45%) presented myxospores, being 9.09% (5/55) only with Myxobolus sp., 5.45% (3/55) only with Henneguya sp., and 70.91% (39/55) presenting both parasites. The presence of myxospores was associated with histological alterations in both stromal and renal parenchyma. Myxospores were found mostly in the peritubular interstitial tissue and in low intensity in the glomerulus which caused nuclear hypertrophy and loss of Bowman space. An increase in the glomerular tuft and a reduction in the lumen of the collector tubules were also observed, besides the high number of melanomacrophage cells in the glomerulus. This study reports for the first time detection of myxozoan mixed infection in one organ of pacu and discuss the possible transportation of myxospores in the circulating blood.

Light and scanning electron microscopic studies of Myxobolus indica n. sp. and a report of three Myxozoan (Myxosporea: Bivalvulida) parasites of cultured ornamental goldfish, Carassius auratus L. for the first time in India

The ornamental fish industry is an economically viable sector in India which suffers from different ectoparasitic infestations, including the myxozoan parasites. An icthyoparasitological survey of myxozoan infections in ornamental fish farms in India revealed the presence of four myxozoan parasites belonging to the family Myxobolidae, in the genera Myxobolus and Thelohanellus. The myxozoan spores were small to large, spherical to ellipsoidal in size. The plasmodia measured 0.5–3.0mm in diameter with disporic pansporoblasts and mature spores. During the survey the authors identified for the first time in India, three previously described species, namely, M. mehlhorni, T. nikolskii and T. batae; and one new species M. indica n. sp., all infecting the ornamental goldfish, Carassius auratus. The present study thus reports a new host, and a new locality for T. batae and M. mehlhorni. The description of T. nikolskii is the first record found in India. The spore of M. indica n. sp. measures 5.8±0.2×4.1±0.5μm in size, having two equal shaped pyriform polar capsules measuring 4.1±0.4×2.7±0.6μm. The results from a combination of light and scanning electron microscopic observations along with a comparison with closely related species were incorporated here. Molecular data is needed to complete the description of the new species.

Myxobolus lepomis n. sp. (Cnidaria: Myxobolidae), a gill myxozoan infecting Lepomis marginatus Holbrook and Lepomis miniatus Jordan (Perciformes: Centrarchidae), in the Big Thicket National Preserve, Texas, USA.

A parasitological survey of freshwater fishes in the Big Thicket National Preserve in southeast Texas revealed myxozoan infections in two species of sunfish, Lepomis marginatus Holbrook and Lepomis miniatus Jordan (Perciformes: Centrarchidae). Pseudocysts were elongate-oval, 988 × 485 µm (ex L. marginatus) and 800 × 606 µm (ex L. miniatus) and demonstrated a predilection to the edge of the primary gill lamellae. Myxospores consistent with the genus Myxobolus were oblong, 16.8-21.3 (19.0 ± 0.9) µm long, 7.0-8.8 (7.9 ± 0.5) µm wide and 5.3-6.1 (5.8 ± 0.3) µm thick (ex L. marginatus) and 17.2-20.3 (18.8 ± 0.7) µm long, 7.5-9.9 (8.7 ± 0.6) µm wide, and 6.8-7.2 (7.0 ± 0.2) µm thick (ex L. miniatus); with 2 pyriform polar capsules 8.3-9.8 (9.0 ± 0.5) µm long, 2.2-2.7 (2.5 ± 0.2) µm wide (ex L. marginatus) and 9.2-10.5 (10.0 ± 0.4) µm long, 2.2-3.0 (2.8 ± 0.2) µm wide (ex L. miniatus). Statistically, the measurements of spore body width, polar capsule length, and polar capsule width were significantly different between myxospores from L. marginatus and L. miniatus. However, intraspecific genetic variability between isolates at the 18S rRNA gene was negligible, with < 0.8% variability across > 2,000 bp of sequence. The isolates shared no significant sequence similarity with any myxozoan deposited in the GenBank nucleotide database. Phylogenetic analysis inferred from the 18S rRNA gene from both L. marginatus and L. miniatus placed the isolates within a clade of myxozoan parasites of perciform fishes. Based on shared tissue and host family tropism, overlapping morphological characters and high degrees of sequence conservation at the 18S rRNA gene, we propose these isolates as morphologically distinct, genetically conspecific representatives of M. lepomis n. sp. from the gills of L. marginatus and L. miniatus in the Big Thicket National Preserve in Texas, USA.

Studies on histopathological alteration of three major organs of the goldfish, Carassius auratus L., of India due to myxozoan infection with special reference to scanning electron microscopic observation.

Severe infections by the protozoan parasite belonging to the genera Myxozoa are causing serious problems in ornamental fish reared in breeding farms. Histopathological study is being used for diagnosis of the severity of diseases. Myxozoan infections result in large scale histopathological damages in different fish tissues. No information is available regarding the histopathological changes of goldfish due to infection with myxozoans. The present study deals with the histopathological changes of the gill, fin, and skin of goldfish, infected with myxozoan parasites along with an ultrastructural study using scanning electron microscope. Several histological alterations have been observed in goldfish, like tissue damage, epithelial hyperplasia, necrosis, anoxia, localized lymphocytic infiltration, excess mucus, cellular necrosis, and epithelial proliferation. The present study revealed the invasion intensified by the occurrence of morphological lesions in the gill, skin, and fin exposed to Myxosporidia, which may lead to respiratory insufficiency in fish and even cause mass mortality.

Myxozoan infection in the muscle layer of the intestine of Rhamdia quelen from the Amazon River Basin, Brazil

ABSTRACT: The present study investigated the morphology and pathology associated with the occurrence of cysts caused by Myxobolus sp. in the intestine of the silver catfish Rhamdia quelen . Comparisons with the other Myxobolus species that infect the intestines of cyprinids and siluriforms revealed that the pyriform spores of Myxobolus sp. are similar in shape to those described previously in M. miyairii, M. duodenalis and M. cunhai, but different in size. Morphometric analyses revealed that mature spores of Myxobolus sp. (10.9 x 5.1μm) are smaller than those of most species except M. cunhai (10.0 x 5.0μm), which has been described infecting the Brazilian catfish Pimelodus clarias , although the spores differ morphologically in relation to the shape of the polar capsules. Further research is necessary in order to clarify the taxonomic and phylogenetic relationships among these congeneric species.

Repatriation of an old fish host as an opportunity for myxozoan parasite diversity: The example of the allis shad, Alosa alosa (Clupeidae), in the Rhine.

BackgroundWildlife repatriation represents an opportunity for parasites. Reintroduced hosts are expected to accumulate generalist parasites via spillover from reservoir hosts, whereas colonization with specialist parasites is unlikely. We address the question of how myxozoan parasites, which are characterized by a complex life-cycle alternating between annelids and fish, can invade a reintroduced fish species and determine the impact of a de novo invasion on parasite diversity. We investigated the case of the anadromous allis shad, Alosa alosa (L.), which was reintroduced into the Rhine approximately 70 years after its extinction in this river system.MethodsWe studied parasites belonging to the Myxozoa (Cnidaria) in 196 allis shad from (i) established populations in the French rivers Garonne and Dordogne and (ii) repatriated populations in the Rhine, by screening the first adults returning to spawn in 2014. Following microscopical detection of myxozoan infections general myxozoan primers were used for SSU rDNA amplification and sequencing. Phylogenetic analyses were performed and cloned sequences were analyzed from individuals of different water sources to better understand the diversity and population structure of myxozoan isolates in long-term coexisting vs recently established host-parasite systems.ResultsWe describe Hoferellus alosae n. sp. from the renal tubules of allis shad by use of morphological and molecular methods. A species-specific PCR assay determined that the prevalence of H. alosae n. sp. is 100 % in sexually mature fish in the Garonne/Dordogne river systems and 22 % in the first mature shad returning to spawn in the Rhine. The diversity of SSU rDNA clones of the parasite was up to four times higher in the Rhine and lacked a site-specific signature of SNPs such as in the French rivers. A second myxozoan, Ortholinea sp., was detected exclusively in allis shad from the Rhine.ConclusionsOur data demonstrate that the de novo establishment of myxozoan infections in rivers is slow but of great genetic diversity, which can only be explained by the introduction of spores from genetically diverse sources, predominantly via straying fish or by migratory piscivorous birds. Long-term studies will show if and how the high diversity of a de novo introduction of host-specific myxozoans succeeds into the establishment of a local successful strain in vertebrate and invertebrate hosts.Graphical Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1760-6) contains supplementary material, which is available to authorized users.

Assessing myxozoan presence and diversity using environmental DNA

Amplicon sequencing on a High Throughput Sequencing platform (custom barcoding) was used to detect and characterise myxosporean communities in environmental DNA samples from marine and freshwater environments and in faeces of animals that may serve as hosts or whose prey may host myxosporean infections. A diversity of myxozoans in filtered water samples and in faeces of piscivores (otters and great cormorants) was detected, demonstrating the suitability of lineage-specific amplicons for characterising otherwise difficult to sample parasite communities. The importance of using this approach was highlighted by the lack of myxosporean detection using commonly employed, broadly targeted eukaryotic primers. These results suggest that, despite being frequently present in eDNA samples, myxozoans have been generally overlooked in “eukaryote-wide” surveys. Lineage-specific primers, in contrast, detected 107 operational taxonomic units that were assigned to both the “freshwater” and “marine” myxosporean lineages. Only 7% of these OTUs clustered with sequences in GenBank, providing evidence for substantial undescribed myxosporean diversity. Many new operational taxonomic units, including those found in otter faeces, clustered with a clade of myxosporeans previously characterised by sequences from invertebrate hosts and water samples only. Because myxozoan species identification is heavily reliant on molecular signatures, lineage-specific amplicon sequencing offers an effective and non-destructive means of improving our knowledge of myxozoan diversity. In addition, the analysis of myxozoan DNA in faeces of piscivores offers a potentially efficient method of sampling for diversity and revealing life cycles as piscivore activities may integrate myxozoan infections in fish over relatively broad spatial scales.

Description of two new species of Myxobolus Bütschli, 1892, M. peleci n. sp. and M. cultrati n. sp., detected during an intensive mortality of the sichel, Pelecus cultratus (L.) (Cyprinidae), in Lake Balaton, Hungary

In the summer of 2014, mass mortality of the sichel, Pelecus cultratus (L.), was observed in Lake Balaton, Hungary. Parasitological examination conducted in the framework of a complete diagnostic survey revealed myxozoan infections. Two species of Myxobolus Bütschli, 1892 were found, one in the gill lamellae and another in the eyes. Following this mass mortality, 113 sichel specimens were examined during a 14-month period. Gill infection with scattered spores in the lamellae was found in 51 fish, while infection in the eyes was recorded in three specimens only. Based upon the morphological and molecular biological data the species from the gills is described here as Myxobolus peleci n. sp. and the species from the eye as M. cultrati n. sp. The 18S rDNA sequences of the two species proved that they differ from all known Myxobolus spp. with sequence data available in the GenBank database. Histological examinations revealed that the spores found in the gill lamellae were derived from plasmodia developing in and around the afferent branchial arteries of the gill arches. No mortality of sichel was recorded in 2015. Infection with these two Myxobolus spp. does not seem to play a role in the mortality of the host fish.

Myxozoan infections of caecilians demonstrate broad host specificity and indicate a link with human activity

Myxozoans are parasitic cnidarians that infect a wide variety of hosts. Vertebrates typically serve as intermediate hosts whereas definitive hosts are invertebrates, including annelids and bryozoans. Myxozoans are known to exploit species in two of the three extant amphibian orders (Anura: frogs and toads; Caudata: newts and salamanders). Here we use museum collections to determine, to our knowledge for the first time, whether myxozoans also exploit the third amphibian order (Gymnophiona: caecilians). Caecilians are a poorly known group of limbless amphibians, the ecologies of which range from aquatic to fully terrestrial. We examined 12 caecilian species in seven families (148 individuals total) characterised by a diversity of ecologies and life histories. Using morphological and molecular surveys, we discovered the presence of the myxozoan Cystodiscus axonis in two South American species (one of seven examined families) of aquatic caecilians – Typhlonectes natans and Typhlonectes compressicauda. All infected caecilians had been maintained in captivity in the United Kingdom prior to their preservation. Cystodiscus axonis is known from several Australian frog species and its presence in caecilians indicates a capacity for infecting highly divergent amphibian hosts. This first known report of myxozoan infections in caecilians provides evidence of a broad geographic and host range. However, the source of these infections remains unknown and could be related to exposure in South America, the U.K. or to conditions in captivity.

Myxozoan infections of the three Indian major carps in fish ponds around Meerut, UP, India, with descriptions of three new species, Myxobolus basuhaldari sp. n., M. kalavatiae sp. n. and M. meerutensis sp. n., and the redescription of M. catlae and M. bhadrensis.

New myxosporean species are described from Indian fishes cultured in pond farms of Meerut, Uttar Pradesh (UP) state. Based upon plasmodia found in the Indian major carps (Catla catla, Cirrhinus cirrhosus, Labeo rohita and their hybrids), three new Myxobolus spp., Myxobolus basuhaldari sp. n., Myxobolus kalavatiae sp. n. and Myxobolus meerutensis sp. n., are described, and two species, Myxobolus catlae and Myxobolus bhadrensis, are redescribed. Plasmodia of M. basuhaldari sp. n., M. kalavatiae sp. n., M. meerutensis sp. n. and M. catlae developed in small cysts in the gill lamellae, while plasmodia and scattered spores of M. bhadrensis were found in the muscles and kidney, respectively. Plasmodia and spores found in these fishes differed from each other with respect to their morphology, tissue tropism and 18S ribosomal DNA (18S rDNA) sequence. No major pathological changes were found, but severe infections were observed.