Evidence of the American Myxobolus dechtiari was introduced along with its host Lepomis gibbosus in Europe: Molecular and histological data.

The American pumpkinseed Lepomis gibbosus, was introduced to Europe more than one hundred years ago. Currently it is a common fish in European freshwaters but relatively few specific parasites infect this fish in this new habitat. In Europe only a single species, Myxobolus dechtiari seems to represent the American myxosporean fauna of centrarchid fishes. M. dechtiari was found in both Portugal and Hungary. This species forms plasmodia with elongated shape inside the cartilaginous rays of gill filaments. In the advanced stage of infection, after disruption of plasmodia, small groups of myxospores remain enclosed in the cartilaginous gill rays causing distortions in the filaments. Myxospores were ellipsoidal in frontal view and lemon-shape in sutural, length 12.5 ± 0.46 (12-13.4) µm, width 10 ± 0.37 (9.6-10.4) µm, and thickness 7.4 ± 0.37 (7-8) µm; the polar capsules were pyriform, equal in size, length 5.6 ± 0.21 (5.3-6) µm, width 3.2 ± 0.16 (3-3.6) µm; Seven to eight polar tube coils were arranged perpendicularly to the capsule length. There was a small, round, 0.4 ± 0.1 (0.3-05) (N = 50) intercapsular appendix in the spores. The small subunit ribosomal DNA (ssrDNA) of M. dechtiari differed from other myxozoans sequenced to date. Phylogenetic analysis of the ssrDNA gene sequence placed this species in a clade including actinospores and Myxobolus species: Raabeia type1, Triactinomyxon sp., and Myxobolus osburni infecting the same host fish. The focus of our study was to prove that the pumpkinseed, a fish originated from North-America introduced one of its myxosporean parasite to Europe. Emphasis was put on to demonstrate the unique feature of this parasite causing infection in the cartilaginous gill rays.

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.

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

INTRODUCTION: 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. OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Malformations of the gill filaments of the ruffe Gymnocephalus cernuus (L.) (Pisces) caused by echinostomatid metacercariae.

In parasite surveys of fishes from Lake Balaton and its tributaries in Hungary, infections with metacercariae of a species of the digenean genus Echinochasmus (Trematoda: Echinostomatidae) were found in seven species of fish. In ruffe, Gymnocephalus cernuus, malformations of the gill filaments apparently caused by these infections were observed. These malformations were in the form of bifurcations of the filaments at about their mid-length. At the point where the filaments bifurcate, an Echinochasmus metacercaria was always embedded in the cartilaginous ray of the gill filament. All specimens of the ruffe were found to be infected by these metacercariae, and each ruffe specimen was infected by 30-300 metacercariae. Such a bifurcation was found in all of the ruffe specimens, but, apart from these gill malformations, the metacercariae produced only local changes in the cartilage. In the other six infected fish species, only local signs were observed in the cartilage. Experimental infections of chicks with metacercariae resulted in the finding of the sexual adult (marita) of an unidentified species of Echinochasmus. ITS sequences of the adult and metacercaria corresponded with each other, and also with a cercaria isolated from a gravel snail (Lithoglyphus naticoides), with a 99.5-100% similarity.

Diversity and activity of cultivable aerobic planktonic bacteria of a saline Lake located in Sovata, Romania.

Aerobic bacterial strains from the salt water of Lake Red (Sovata, Romania) were cultivated. More than half of the 80 strains were G(-) and formed motile straight rods. Only a few strains produced acid from D-glucose and reduced nitrate to nitrite. Optimum NaCl concentration for growth varied between 5 and 15 % in the majority of the strains, so the isolates were regarded moderately halophilic. On the basis of the 16S rRNA gene sequence similarity almost half of the strains were identified as members of genus Halomonas. Other strains belonged to genera Marinobacter, Psychrobacter, Serratia, Morganella (γ-Proteobacteria), Bacillus, Exiguobacterium, Planococcus (Firmicutes), and Arthrobacter, Micrococcus, Microbacterium, and Nesterenkonia (Actinobacteria).

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.

Infection of the heart of the common bream, Abramis brama (L.), with Myxobolus s.l. dogieli (Myxozoa, Myxobolidae).

Myxobolus dogieli Bykhovskaya-Pavlovskaya & Bykhovski, 1940 is regarded as a site specific myxosporean, infecting the heart of cyprinid fish. During a survey of the myxosporean fauna of Lake Balaton fish, heart myxobolosis was found in the common bream, Abramis brama, with heavy infection of the ventricle and the bulbus arteriosus in some infected bream. Developing and mature plasmodia were mostly in the connective tissue of the subepicardium and subendocardium. Plasmodia developing in the subendocardium protruded into the lumen of the heart, while plasmodia developing in the subepicardium protruded over the epicardium forming large sausage-like outgrowths. Plasmodia with mature spores were found in the summer. The shape and size of the spores corresponded to those of the original description. Phylogenetic analysis based on the 18S rDNA sequence of M. dogieli showed that this species fit well in the genus Myxobolus. As no molecular data are available on spores from the type host, common carp, the species studied by us is temporarily designated as Myxobolus s.l. dogieli.

Occurrence and regional distribution of SHV-type extended-spectrum beta-lactamases in Hungary.

In order to determine the presence and geographical distribution of SHV-type extended-spectrum beta-lactamase genes among Enterobacteriaceae strains in Hungary, isolates from 25 microbiology laboratories throughout the country were collected between January 2002 and August 2003 and examined. Sequencing of the genes showed that SHV-5 and SHV-2a are the dominant SHV-types in extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in this country. The SHV-2 gene, which is prevalent in many European countries, was not detected, but one isolate carried the SHV-12 gene. The results show that these genes are circulating among Enterobacteriaceae strains in Hungary and indicate that strict infection control measures are warranted in order to prevent their spread.