The occurrence of known Myxobolus and Thelohanellus species (Myxozoa, Myxosporea) from Indian major carps with the description of Myxobolus bandyopadhyayi n. sp. in West Bengal.

Myxosporean infection of Indian major carps (rohu, Labeo rohita; catla, Gibelion catla; mrigal, Cirrhinus mrigala) was examined from two fish farms and two fish markets in West Bengal, India. One Thelohanellus and four Myxobolus species were detected from the fins and scales of the investigated species. Comprehensive morphological and molecular biological studies revealed four already known species, Thelohanellus caudatus from the fins of rohu, Myxobolus dermiscalis from the scales of rohu, Myxobolus chakravartyi from the fins of catla, and Myxobolus rewensis from the fins of mrigal. This study complemented the species description of M. chakravartyi and M. rewensis with the missing molecular data. Moreover, based on morphometrics and ssrDNA sequence data, a new species was documented from the scales of rohu, and named Myxobolus bandyopadhyayi n. sp.

New data on Thelohanellus nikolskii Achmerov, 1955 (Myxosporea, Myxobolidae) a parasite of the common carp (Cyprinus carpio, L.): The actinospore stage, intrapiscine tissue preference and molecular sequence.

Thelohanellus nikolskii, Achmerov, 1955 is a well-known myxozoan parasite of the common carp (Cyprinus carpio L.). Infection regularly manifests in numerous macroscopic cysts on the fins of two to three month-old pond-cultured carp fingerlings in July and August. However, a Thelohanellus infection is also common on the scales of two to three year-old common carp in ponds and natural waters in May and June. Based on myxospore morphology and tissue specificity, infection at both sites seems to be caused by the same species, namely T. nikolskii. This presumption was tested with molecular biological methods: SSU rDNA sequences of myxospores from fins of fingerlings and scales of older common carp were analysed and compared with each other and with related species available in GenBank. Sequence data revealed that the spores from the fins and scales represent the same species, T. nikolskii. Our study revealed a dichotomy in both infection site and time in T. nikolskii-infections: the fins of young carp are infected in Summer and Autumn, whereas the scales of older carp are infected in Spring. Myxosporean development of the species is well studied, little is known, however about the actinosporean stage of T. nikolskii. A previous experimental study suggests that aurantiactinomyxon actinospores of this species develop in Tubifex tubifex, Müller, 1774. The description included spore morphology but no genetic sequence data (Székely et al., 1998). We examined >9000 oligochaetes from Lake Balaton and Kis-Balaton Water Reservoire searching for the intraoligochaete developmental stage of myxozoans. Five oligochaete species were examined, Isochaetides michaelseni Lastochin, 1936, Branchiura sowerbyi Beddard, 1892, Nais sp., Müller, 1774, Dero sp. Müller, 1774 and Aelosoma sp. Ehrenberg, 1828. Morphometrics and SSU rDNA sequences were obtained for the released actinospores. Among them, from a single Nais sp., the sequence of an aurantiactinomyxon isolate corresponded to the myxospore sequences of T. nikolskii.

Molecular detection of a novel cyprinid herpesvirus in roach (Rutilus rutilus) and asp (Leuciscus aspius) showing typical signs of carp pox disease.

In the early spring of 2018, in Lake Balaton (Hungary), a roach (Rutilus rutilus) and an asp (Leuciscus aspius) were found in an fish trap at the outlet of the river Sió showing typical signs of the so-called carp pox disease, such as foci of epidermal hyperplasia on the head and the whole body surface, including the fins. Molecular tests revealed the presence of the DNA of an unknown fish herpesvirus. Three genes encoding the DNA-dependent DNA polymerase, major capsid protein and ATPase subunit of terminase were amplified and sequenced from the alloherpesviral genome. The gene sequences of the viruses obtained from the two different fish species shared 94.4% nucleotide sequence identity (98.1% amino acid sequence identity), suggesting that they belong to the same virus species. Phylogenetic analysis based on the DNA polymerase (and the concatenated sequences of the amplified genes, as well) implied that the detected virus belongs to the genus Cyprinivirus within the family Alloherpesviridae. The sequences of the novel alloherpesvirus diverge from those of the five cyprinivirus species described previously, so it putatively represents the sixth virus species in the genus.

Detection of cyprinid herpesvirus 1 (CyHV-1) in barbel (Barbus barbus): First molecular evidence for the presence of CyHV-1 in fish other than carp (Cyprinus carpio).

Two adult barbels (Barbus barbus) with visible skin tumours were subjected to histopathological and molecular examinations. The fish were caught in the River Danube near Budapest. Papillomas were found around their oral cavity, at the operculum and at the pectoral fins, while epidermal hyperplasias were seen on the body surface. Cyprinid herpesvirus 1 (CyHV-1) was detected in the kidney of the specimens by polymerase chain reaction (PCR), and barbel circovirus 1 (BaCV1) was found in all internal organs and in the tissues of the tumours. The whole genome of BaCV1 and three conserved genes from the genome of CyHV-1 were sequenced. Previously, BaCV1 had been reported only once from a mass mortality event among barbel fry. The whole genome sequence of our circovirus shared 99.9% nucleotide identity with that of the formerly reported BaCV1. CyHV-1 is known to infect common carp and coloured carp (Cyprinus carpio), and has been assumed to infect other cyprinid fish species as well. We found the nucleotide sequences of the genes of CyHV-1 to be identical in 98.7% to those of the previous isolates from carp. To the best of our knowledge, this is the first molecular confirmation of the presence of CyHV-1 DNA in cyprinid fish species other than carp.

Description of Henneguya jaczoi sp. n. (myxosporea, myxobolidae) from Perca fluviatilis (L.) (pisces, percidae) with some remarks on the systematics of Henneguya spp. of european fishes.

A new Henneguya species, H. jaczoi sp. n., is described from perch (Perca fluviatilis) from Lake Balaton, Hungary. This species infects the palatal region of the fish, forming large plasmodia in the thickened caudal part of the buccal cavity and at the dorsal ends of the cartilaginous gill arches. The species differs from the gill-dwelling Henneguya species of perch and pike (Esox lucius) both morphologically and in molecular aspects. The authors conclude that the type species H. psorospermica Thélohan is a specific parasite of pike, while the species forming plasmodia in the gills of perch corresponds to H. texta Cohn, which was hitherto regarded as a synonym of H. psorospermica. Besides the above-mentioned species, H. creplini was frequently found in pikeperch (Sander lucioperca) and Volga pikeperch (Sander volgensis), but no Henneguya infection has been recorded in ruffe (Gymnocephalus cernua), which is a common percid fish of the lake and is known to be the type host species for H. creplini.

Myxobolus infection in the cornea of the roach (Rutilus rutilus) in Lake Balaton.

Infection of the cornea in fishes by Myxobolus plasmodia is a common but still little known site preference of myxosporeans. A sporadic but striking infection in the cornea of the roach (Rutilus rutilus) was observed in Lake Balaton, Hungary. Relatively small, round plasmodia 250 to 500 µm in diameter developed in the dense connective tissue of the cornea. Morphological and molecular biological examination of spores collected from cysts in the cornea demonstrated that this infection is caused by Myxobolus fundamentalis, a species hitherto reported only from the cartilaginous gill arch of the roach. The 18S rDNA sequences of spores from the cornea showed 99.9% identity to the sequences of spores from the gill arch, and they also shared 99.9% identity with the sequences of triactinomyxon actinospores obtained from the oligochaete Isochaetides michaelseni.

Complete genome sequence and analysis of ictalurid herpesvirus 2.

Ictalurid herpesvirus 2 (IcHV-2) has been causing substantial losses in the black bullhead aquaculture industry since the 1990s. Using next-generation sequencing, the genome of IcHV-2 was completely sequenced and analysed in this study. The complete genome was found to be 142,925 bp in size, containing 77 predicted protein-coding regions, including 12 ORFs that appear to have a homologue in every alloherpesvirus genome sequenced to date. The genome organization of the IcHV-2 shows high similarity to that of IcHV-1, the founding member of the genus Ictalurivirus within the family Alloherpesviridae. A unique sequence region of 101 kbp is flanked by terminal direct repeats of 20 kbp. Thirteen of the 77 putative genes do not show homology to any known genes with sequences in public databases; six of them are found in the repeat regions. Analysis of the whole genome confirms the previously established taxonomic position of IcHV-2.

Molecular detection and genome analysis of circoviruses of European eel (Anguilla anguilla) and sichel (Pelecus cultratus).

The prevalence and distribution of piscine circoviruses (CVs) were tested in a routine virus monitoring programme in Lake Balaton, Hungary. A high prevalence of European eel CV (EeCV) was found in the apparently healthy eel population (35.5%). The copy number of the viral DNA in different organs was determined by quantitative real-time PCR. The results suggested that some eel specimens were in active viraemic status despite their asymptomatic condition. Furthermore, a novel, previously undescribed CV was also detected in eel and sichel samples. Full genome characterisation confirmed that the virus represents a novel EeCV species (EeCV-2). The genome contains an integrated eel chromosome-derived fragment, suggesting that the original host of the virus was the eel and it probably emerged subsequently in the sichel by host switching. In some samples, an additional, 1,111-nt-long circular ssDNA was also observed involving a CV-like stem-loop structure and an ORF showing homology to CV capsid protein genes, without any sign of a replication initiator protein sequence.

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.

Three new species of Myxobolus Bütschli, 1882 (Myxozoa: Myxobolidae) infecting the common nase Chondrostoma nasus (L.) in the River Danube.

The common nase Chondrostoma nasus (L.) is a frequent cyprinid fish in the River Danube. In a survey on its infection with myxosporeans, eight different Myxobolus spp. spore types were found in the gills, swim bladder, fins and intestinal wall. Of these, spore types representing three species were studied in detail by morphological and molecular methods. Based on the differences in 18S rDNA sequences, two new species of Myxobolus Bütschli, 1882 from the gills and one from the swim bladder are described: M. arrabonensis n. sp., M. szentendrensis n. sp. and M. paksensis n. sp. The new species resembled M. muelleri Bütschli, 1882, M. intimus Zaika, 1965 and M. cycloides Gurley, 1893, all parasitic in leuciscine cyprinids, in spore size and location in the host, but exhibited differences in partial 18S rDNA sequences as follows: M. arrabonensis - M. muelleri (1.4%), M. szentendrensis - M. intimus (2.8%), M. paksensis - M. cycloides (2.4%). Based on the significant differences in rDNA sequences, the three forms are considered to represent new, hitherto undescribed species in spite of their morphological similarities to some Myxobolus spp. forming spores in identical locations in genetically closely related cyprinids of the subfamily Leuciscinae.

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.

Whole-genome sequencing of a green bush viper reovirus reveals a shared evolutionary history between reptilian and unusual mammalian orthoreoviruses.

In this study, we sequenced the whole genome of a reovirus isolated from a green bush viper (Atheris squamigera). The bush viper reovirus shared several features with other orthoreoviruses, including its genome organization. In phylogenetic analysis, this strain was monophyletic with Broome virus and baboon orthoreovirus, indicating that these viruses might have originated from a common ancestor. These new molecular data supplement previous information based mainly on biological properties of reptilian reoviruses, confirming their taxonomic position and broadening our knowledge of the evolution of members of the genus Orthoreovirus.

Zoonotic transmission of reassortant porcine G4P[6] rotaviruses in Hungarian pediatric patients identified sporadically over a 15 year period.

Genotype G4P[6] Rotavirus A (RVA) strains collected from children admitted to hospital with gastroenteritis over a 15 year period in the pre rotavirus vaccine era in Hungary were characterized in this study. Whole genome sequencing and phylogenetic analysis was performed on eight G4P[6] RVA strains. All these RVA strains shared a fairly conservative genomic configuration (G4-P[6]-I1/I5-R1-C1-M1-A1/A8-N1-T1/T7-E1-H1) and showed striking similarities to porcine and porcine-derived human RVA strains collected worldwide, although genetic relatedness to some common human RVA strains was also seen. The resolution of phylogenetic relationship between porcine and human RVA genes was occasionally low, making the evaluation of host species origin of individual genes sometimes difficult. Yet the whole genome constellations and overall phylogenetic analyses indicated that these eight Hungarian G4P[6] RVA strains may have originated by independent zoonotic transmission, probably from pigs. Future surveillance studies of human and animal RVA should go parallel to enable the distinction between direct interspecies transmission events and those that are coupled with reassortment of cognate genes.

Photobiology of sea ice algae during initial spring growth in Kangerlussuaq, West Greenland: insights from imaging variable chlorophyll fluorescence of ice cores.

We undertook a series of measurements of photophysiological parameters of sea ice algae over 12 days of early spring growth in a West Greenland Fjord, by variable chlorophyll fluorescence imaging. Imaging of the ice-water interface showed the development of ice algae in 0.3-0.4 mm wide brine channels between laminar ice crystals in the lower 4-6 mm of the ice, with a several-fold spatial variation in inferred biomass on cm scales. The maximum quantum yield of photosynthesis, F(v) /F(m), was initially low (~0.1), though this increased rapidly to ~0.5 by day 6. Day 6 also saw the onset of biomass increase, the cessation of ice growth and the time at which brine had reached <50 psu and >-2 °C. We interpret this as indicating that the establishment of stable brine channels at close to ambient salinity was required to trigger photosynthetically active populations. Maximum relative electron transport rate (rETR(max)), saturation irradiance (E(k)) and photosynthetic efficiency (α) had also stabilised by day 6 at 5-6 relative units, ~30 µmol photons m⁻² s⁻¹ and 0.4-0.5 µmol photons m⁻²s⁻¹, respectively. E(k) was consistent with under-ice irradiance, which peaked at a similar value, confirming that daytime irradiance was adequate to facilitate photosynthetic activity throughout the study period. Photosynthetic parameters showed no substantial differences with depth within the ice, nor variation between cores or brine channels suggesting that during this early phase of ice algal growth cells were unaffected by gradients of environmental conditions within the ice. Variable chlorophyll fluorescence imaging offers a tool to determine how this situation may change over time and as brine channels and algal populations evolve.