Comparison of the Myxobolus fauna of common barbel from Hungary and Iberian barbel from Portugal.

We compared Myxobolus infection of common barbel Barbus barbus from the Danube River in Hungary with that in Iberian barbel Luciobarbus bocagei from the Este River in Portugal. In Hungary, we recorded 5 known Myxobolus species (M. branchialis, M. caudatus, M. musculi, M. squamae, and M. tauricus) and described M. branchilateralis sp. n. In Portugal we recorded 6 Myxobolus species (M. branchialis, M. branchilateralis sp. n., M. cutanei, M. musculi, M. pfeifferi, and M. tauricus). Species found in the 2 habitats had similar spore morphology and only slight differences were observed in spore shape or measurements. All species showed a specific tissue tropism and had a definite site selection. M. branchialis was recorded from the lamellae of the gills, large plasmodia of M. branchilateralis sp. n. developed at both sides of hemibranchia, M. squamae infected the scales, plasmodia of M. caudatus infected the scales and the fins, and M. tauricus were found in the fins and pin bones. In the muscle, 3 species, M. musculi, M. pfeifferi and M. tauricus were found; however they were found in distinct locations. Plasmodia of M. musculi developed intracellularly in muscle cells, plasmodia of M. tauricus were found in the dense connective tissue of the pin bones, whereas M. pfeifferi formed plasmodia in the connective tissue of the intramuscular septa. This latter species was often found in the cartilaginous gill arch as well. Comparative morphological and phylogenetic studies, as well as 18S rDNA sequences, revealed differences between the Myxobolus fauna of the 2 barbel species originating from different geographic regions.

Induced oogenesis of the European eel (Anguilla anguilla L.) in freshwater condition.

European eel is a catadromous fish species, which means that after living in freshwater premature individuals adapt to sea water, and migrate to the Sargasso Sea for spawning. Although male eel can be sexually matured even in freshwater, to date, it was believed that female eel can be matured only in seawater. Here we show that the process of sexual maturation may be induced in freshwater by treating female eels with carp pituitary (GSI = 9.87 ± 1.55%). It is thus proposed that seawater condition is not an obligatory environment for stimulating gametogenesis and for artificial maturation of the European eel in neither gender.

Complete life cycle of Myxobolus rotundus (Myxosporea: Myxobolidae), a gill myxozoan of common bream Abramis brama.

The life cycle of Myxobolus rotundus Nemeczek, 1911, a myxosporean parasite of the gills of common bream Abramis brama L., was studied under laboratory conditions. Mature Myxobolus spp. spores from plasmodia in the gills of wild bream were used to infect naïve oligochaete worms in a flow-through system of aquaria. Triactinomyxon-type actinospores were released from the oligochaetes 1 yr later and allowed to continually flow into a tank containing uninfected bream fry. The gills of the fry were checked for development of plasmodia in squash preparations 3 d postexposure, and then at weekly intervals for 8 wk. Tissue samples were fixed at each time point. Developing plasmodia were first observed 17 d post-exposure (Day 17). Mature spores were collected from plasmodia on Day 56 and were added to plastic dishes containing parasite-free Tubifex tubifex oligochaetes. Second-generation actinospores were released from these worms 8 mo post-exposure, and were morphologically identical to first-generation spores. Myxospores obtained from the bream fry were morphologically identical to those identified in wild bream as M. rotundus. Small subunit ribosomal RNA gene sequences obtained from first- and second-generation actinospores and the bream fry myxospores were 100% similar to M. rotundus spores from the original wild fish.

Myxobolus erythrophthalmi sp. n. and Myxobolus shaharomae sp. n. (Myxozoa: Myxobolidae) from the internal organs of rudd, Scardinius erythrophthalmus (L.), and bleak, Alburnus alburnus (L.).

During a survey of myxosporean parasites of cyprinid fish in Hungary, infections caused by unknown Myxobolus spp. were found in the internal organs of rudd, Scardinius erythrophthalmus, and bleak, Alburnus alburnus. Small plasmodia developed in blood vessels of the kidney, liver, testes and intestinal wall. The parasites were studied on the basis of spore morphology and by histological and molecular methods. In most cases, plasmodia were surrounded by host tissue without a host reaction; however, in advanced cases, a connective tissue capsule was seen around plasmodia. Spores collected from the two fish species differed from each other and from the known Myxobolus spp. both in their morphology and 18S rDNA sequences. The two species, described as M. erythrophthalmi sp. n. from rudd and M. shaharomae sp. n. from bleak, are characterized by a specific histotropism to blood vessels, while the organ specificity involves the kidney and for the latter species, most internal organs.

Myxozoan infections in fishes of the Tasik Kenyir Water Reservoir, Terengganu, Malaysia.

During a survey on fishes of the Tasik Kenyir Reservoir, Malaysia, 5 new Myxobolus spp. and 2 known Henneguya spp. were found. The specific locations for 2 Myxobolus spp. were the host's muscles, while 2 other Myxobolus spp. were found to develop in the host's kidney and gills, respectively. Of the species developing intracellularly in muscle cells, M. terengganuensis sp. nov. was described from Osteochilus hasselti and M. tasikkenyirensis sp. nov. from Osteochilus vittatus. M. csabai sp. nov. and M. osteochili sp. nov. were isolated from the kidney of Osteochilus hasselti, while M. dykovae sp. nov. was found in the gill lamellae of Barbonymus schwanenfeldii. Henneguya shaharini and Henneguya hemibagri plasmodia were found on the gills of Oxyeleotris marmoratus and Hemibagrus nemurus, respectively. Description of the new and known species was based on morphological characterization of spores, histological findings on locations of plasmodia and DNA sequence data.

Myxozoan pathogens of Malaysian fishes cultured in ponds and net-cages.

We describe new myxosporean species from Malaysian fishes cultured in pond farms and net-cages. Myxobolus omari sp. nov. and M. leptobarbi sp. nov. were found in the muscles of Pangasianodon hypophthalmus and Leptobarbus hoevenii, respectively, while plasmodia and spores of Thelohanellus zahrahae sp. nov. and Henneguya daoudi sp. nov. were detected in the gills of Barbonymus gonionotus and Trichogaster trichopterus, respectively. Plasmodia and spores found in these fishes differed from the known myxosporean species in respect of their morphology, tissue tropism and 18S rDNA structure. No major pathological changes were found, but in the future these species might pose a potential threat to more intensified fish culture.

The role of copepods (Cyclops spp.) in eliminating the actinospore stages of fish-parasitic myxozoans.

The actinospore consumption of copepods (Cyclops spp.) was demonstrated by laboratory observations. It was observed that in experimental dishes the number of actinospores floating in the water decreased, or such actinospores were completely eliminated, in the presence of copepods. The ingestion of actinospores by copepods and their further fate were monitored by fluorescent staining and by conventional histological techniques. The actinospores were observed to have got caught on the filters of Cyclops spp. Two and a half hours after the copepods had been placed into water containing actinospores, their digestive tract was found to contain spores that had extruded their filaments from the polar capsules. After copepods having ingested the actinospores of the species Myxobolus pseudodispar had been fed to roaches, no muscle infection developed in the fish host. It is likely that Cyclops spp. can filter out actinospores floating in the water also from natural waters, thus decreasing the chance of development of myxosporean infections.

Comparative morphological and molecular studies on Myxobolus spp. infecting chub from the river Danube, Hungary, and description of M. muellericus sp. n.

During a survey on fishes from the River Danube, the occurrence of 8 Myxobolus species (Myxozoa: Myxobolidae) was registered in chub Leuciscus cephalus L. Most species had a specific location within the fish host. M. cycloides was found in the wall of the swimbladder; the branched plasmodia of M. dujardini were located typically in the epithelium of the non-lamellar part of gill filaments; the plasmodia of M. ellipsoides infected fins between 2 fin rays; M. muelleri and Myxobolus sp. 2 formed large elongated plasmodia in the afferent gill artery of filaments, while the round cysts of M. muellericus sp. n. filled the capillary network of the gill lamellae. Intramuscular plasmodia of M. pseudodispar proved to be the most common, although large cysts of Myxobolus sp. 1 were also frequently found in the intestinal wall. Despite similarities of some species in spore morphology, 18S rDNA sequences showed clear differences between the species examined.

Description of a new synactinomyxon type from the River Sousa, Portugal.

Actinospore infection of oligochaetes collected from the mud of 2 freshwater biotopes in Portugal was studied. Using the 'cell-well plate method', a new synactinomyxon type was found in 2 specimens (1.3%) of the examined Tubifex tubifex oligochaetes from the River Sousa north of Porto, Portugal. In Criodrilus lacuum and Dero digitata specimens collected from the same river, no actinosporeans were released during the 12 wk observation period. Infected oligochaetes were only found immediately post-collection, and no further actinosporean release was recorded in Tubifex specimens kept alive for several weeks. Actinospore infection showed high intensity in oligochaetes in both positive cases. No actinosporean stages of myxosporeans have as yet been described from Portugal. On the basis of spore morphology and 18S rDNA sequence data, the synactinomyxon type presented in this paper differs from those already known and described in the literature.

Molecular phylogeny of the kidney-parasitic Sphaerospora renicola from common carp (Cyprinus carpio) and Sphaerospora sp. from goldfish (Carassius auratus auratus).

The genetic relatedness of two kidney-parasitic Sphaerospora species was studied. Although S. renicola, the causative agent of swimbladder inflammation of common carp fingerlings (Cyprinus carpio), and Sphaerospora sp. originating from goldfish (Carassius auratus auratus) were indistinguishable on the basis of spore morphology, they were found to be genetically different as their 18S rDNA sequences shared only 71.9% identical nucleotides. In the phylogenetic trees, Sphaerospora sp. from goldfish grouped with Myxidium truttae (AJ582061) within the clade of the coelozoic freshwater species. Sphaerospora renicola clustered with S. molnari (AF378345) within the group of myxosporeans histozoic in gills. The topology of the six Sphaerospora species on the phylogenetic trees implied that myxospore morphology does not correlate with the genetic relationships, and the genus seems to be polyphyletic.

Occurrence and pathology of Sinergasilus lieni (Copepoda: Ergasilidae), a parasite of the silver carp and bighead, in Hungarian ponds.

Sinergasilus lieni Yin, 1949, a well-known and pathogenic parasitic copepod in China and Russia, has been detected in Hungarian carp farms for the first time. The parasite infected the third-year generation of silver carp (Hypophthalmichthys molitrix) and bighead (Aristichthys nobilis). The gills of the infected fish specimens showed severe pathological changes. At the attachment sites of female copepods clubbing and fusing of the gill filaments were observed and in some parts of the pale or whitish hemibranchia deep indentations were recorded in places where the tips of the damaged filaments had broken off. Silver carp and bighead were infected at a similar rate, having 8 to 27 copepods attached to the end of the clubbed filaments or the proliferated epithelium of 2 to 10 fused filaments. In histological sections the head part of the parasite was found in a deep cavity of the proliferated epithelium, piercing its antennae deep into the tissues. Only the end of the filaments showed changes. In this part the proliferated epithelium was infiltrated by eosinophilic granular cells. In the central and basal parts of the hemibranchia the original structure of the filaments was preserved with intact secondary lamellae.

Reno-, hepato- and splenomegaly of common carp fingerlings (Cyprinus carpio L.) diseased in swimbladder inflammation caused by Sphaerospora renicola Dyková et Lom, 1982.

The weight of internal organs (swimbladder, kidney, liver, spleen) in relation to the body weight was studied in common carp fingerlings divided into three groups on the basis of swimbladder appearance and microscopic examination of the kidney. The fish had been collected from different Hungarian fish farms at the time when swimbladder inflammation (SBI) usually occurs (in July and August). The first group comprised fish with severe signs of SBI and massive renal sphaerosporosis, the second group consisted of fish with milder swimbladder changes and/or kidney infection by a low number of Sphaerospora renicola, while the third group was constituted by infection-free common carp fry. Statistical analysis of swimbladder, kidney, liver and spleen weight in relation to the body weight revealed that in the infected groups the internal organs were substantially enlarged. This suggests that in common carp fry with SBI the swimbladder changes are accompanied by reno-, hepato- and splenomegaly.

A survey of coccidian infections of freshwater fishes of Peninsular Malaysia, with descriptions of three species of Goussia Labbé, 1896 (Apicomplexa: Eimeriidae).

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.

Development of Myxobolus macrocapsularis (Myxosporea: Myxobolidae) in an oligochaete alternate host, Tubifex tubifex.

The development of Myxobolus macrocapsularis Reuss, 1906, a myxosporean parasite of the gills of common bream Abramis brama L., was studied in experimentally infected oligochaetes. In 3 experiments uninfected Tubifex tubifex Muller and Limnodrilus hoffmeisteri (Claparéde) were exposed to mature myxospores of M. macrocapsularis. In all experiments, typical triactinospores developed in T. tubifex specimens but no infection was found in L. hoffmeisteri. Triactinospores were released from oligochaetes 66 to 99 d after initial exposure. At that time pansporocysts containing 8 triactinospores were located in the gut epithelium of experimental oligochaetes, but free actinosporean stages were also found in the gut lumen of the oligochaetes. Each triactinospore had 3 pyriform polar capsules and a barrel-shaped sporoplasm with 32 secondary cells. The spore body joined the 3 caudal projections with a stout style.