Huffmanela paronai sp. n. (Nematoda: Trichosomoididae), a new parasite from the skin of swordfish Xiphias gladius in the Ligurian sea (Western Mediterranean)

A new species of trichosomoidid nematode, Huffmanela paronai sp. n., is established on the basis of its egg morphology and biological characters. The dark-shelled, embryonated eggs of this histozoic parasite occur in masses in the epidermis of the swordfish Xiphias gladius L. (Xiphiidae, Perciformes) from the Ligurian Sea in northern Italy. The eggs are concentrated in groups appearing as black spots in the skin of the fish host, being distributed mainly on the lower part of its body (lower jaw, gill covers, pectoral, anal and caudal fins, lower half of body). The parasite's eggs are characterised mainly by their shape and markedly small size (48-51 x 21-24 microm), an aspinose surface, relatively small polar plugs, and thick egg wall (3 microm). This is the first Huffmanela species reported from fish in Europe.     

Intraoligochaete development of Myxobolus intimus (Myxosporea: Myxobolidae), a gill myxosporean of the roach (Rutilus rutilus)

The infection with Myxobolus intimus Zaika, 1965 in the gills of the roach Rutilus rutilus (L.) from Lake Balaton was recorded in 28 out of the 39 fish examined. Developing and mature plasmodia were detected on the gills exclusively in the spring. The Myxobolus intimus infection was found only in 2- to 3-year-old fish. In histological sections, young plasmodia were found in capillaries of the secondary lamellae. More mature, round plasmodia 0.4-0.6 mm in diameter, deformed the respiratory lamellae. The intraoligochaete development of M. intimus was studied in experimentally infected oligochaetes. In two experiments, uninfected Tubifex tubifex Müller and Limnodrilus hoffmeisteri (Claparède) were exposed to mature myxospores of M. intimus. In both experiments, typical triactinospores developed in T. tubifex specimens but no infection was found in L. hoffmeisteri. In semithin sections, developmental stages, pansporocysts and actinospores, were found within the proliferated gut epithelium of T. tubifex. Triactinospores were first released from oligochaetes 37 and 58 days after initial exposure in the two experiments, respectively. Each triactinospore had three pyriform polar capsules and a cylindrical sporoplasm with 32 secondary cells. The spore body joined the 3 caudal projections with a moderately long style.     

Myxozoans infecting the sharptooth catfish, Clarias gariepinus in the Okavango River and Delta, Botswana, including descriptions of two new species, Henneguya samochimensis sp. n. and Myxobolus gariepinus sp. n

During a recent investigation of parasites infecting fishes from the Okavango River and Delta, Botswana (southern Africa) fourteen sharptooth catfish, Clarias gariepinus (Burchell, 1822) (Siluriformes: Clariidae) were examined for the presence of myxozoan infections. Results revealed the presence of two species of the genus Henneguya Thélohan, 1895 and one species of the genus Myxobolus Bütschli, 1882 infecting this fish host. Two of the sampled fish exhibited large plasmodia of Henneguya suprabranchiae Landsberg, 1987 in the cartilage of the accessory breathing organ, another two individuals were infected with H. samochimensis sp. n. plasmodia in the gills and another three individuals revealed an infection with Myxobolus gariepinus sp. n. plasmodia in the ovaries.     

The development of Myxobolus pavlovskii (Myxozoa: Myxobolidae) includes an echinactinomyxon-type actinospore

Echinactinomyxon-type actinospores were found in a mixed-species oligochaete culture originating from the Temperate Water Fish Hatchery near Budapest, Hungary. On the basis of DNA sequence analysis, the actinospores were identified as Myxobolus pavlovskii (Akhmerov, 1954), the 18S rDNA sequence from myxospores of which is available in GenBank. Silver carp Hypophthalmichthys molitrix (Valenciennes) fry specimens were successfully infected by cohabitation with the echinactinomyxon-releasing oligochaetes, which confirmed the molecular data congruence. The echinactinomyxons and the myxospores that developed in the gills of exposed fish fry were analysed morphologically and on DNA basis. The infected gill tissue was examined histologically. As typical characters of M. pavlovskii, numerous small plasmodia were observed in the epithelia of gill lamellae. Plasmodia contained thousands of myxospores with polar capsules unequal in size and witl; large intercapsular processes. The 18S rDNA sequence from actinospores and those from myxospores originating from the experimentally infected fish were identical. The oligochaete species releasing actinospores was morphologically determined as Limnodrilus sp. This is the first record of an echinactinomyxon as an alternate stage within the genus Myxobolus.     

Phylogeny of Coccomyxa (Myxosporea: Myxidiidae) spp. with the description of a new species from Bathygobius cyclopterus (Gobiidae) in the northern Red Sea

Two species of Coccomyxa Léger et Hesse, 1907, one of the least studied myxosporean genera, are reported from shallow coastal waters in the Gulf of Eilat, Red Sea, Israel. A new species, Coccomyxajirilomi sp. n. is described from the spotted frillgoby Bathygobius cyclopterus (Valenciennes) (Gobiidae). It forms polysporous plasmodia that invade the liver and form packed clusters inside the bile ductules. Plasmodia also occur in the bile ducts and gall bladder of the host, attached to the epithelial lining or free floating in the bile. Infected hepatic bile ductules packed with plasmodia were partially occluded, with evidence of cholestasis, periductular fibrosis and pericholangitis. The mature spore is ellipsoid, has smooth valves and contains a single polar capsule with the polar filament arranged in 4-5 oblique coils. Spore dimensions are 9.0-11.3 x 5.0-7.0 microm. A second species, Coccomyxa sp., with smaller 7.6-9.6 x 4.2-5.2 microm and more delicate spores, was found in the gall bladder of the rippled rockskipper, Istiblennius edentulus (Forster et Schneider) (Blenniidae). The small subunit (SSU) rDNA sequence analysis of both Coccomyxa species suggests that they are closely related to members of the genera Myxidium, Zschokkella and Auerbachia, whose members infect the gall bladder of marine fish.     

Temporal and spatial distribution of glochidial larval stages of European unionid mussels (Mollusca: Unionidae) on host fishes

Glochidia are the larval stage of freshwater unionid mussels that parasitize the fins and gill apparatus of fish. A total of 22 fish species were examined for the presence of glochidia whose distribution on individual hosts was studied on three common fish species, the roach Rutilus rutilus (L.), perch Percafluviatilis L. and bitterling Rhodeus sericeus (Pallas). Between 1997 and 1999, the fish were obtained from the rivers Morava and Kyjovka and surrounding water pools in the Czech Republic. The glochidia of two genera, Unio and Anodonta, were found. Anodonta glochidia were observed on 10 fish species, Unio glochidia on 17 fish species. There was a difference in spatial distribution of glochidia on the body of the host fish. Unio glochidia were predominantly located on the gills, whereas most Anodonta glochidia were found on the fins, with the highest numbers of glochidia were observed on the margin of the pectoral fins. For the gill apparatus, Unio glochidia were found predominantly on the second and third arch. Anodonta glochidia were predominantly found during winter and spring (November-May), whereas Unio glochidia were more abundant during May and June. The number of glochidia was positively correlated with fish length in perch highly infected by Anodonta glochidia and perch infected by Unio glochidia. Of the three fish species, the highest occurrence of parasites was found on perch with fewer observed on roach. In spite of the close relationship between bitterling and unionid mussels, glochidiosis was rare on this fish species.     

Fine structure of Henneguya pilosa sp. n. (Myxozoa: Myxosporea), parasite of Serrasalmus altuvei (Characidae), in Brazil

Henneguya pilosa sp. n., a new species of myxosporean from the gill filaments of the white piranha, Serrasalmus altuvei Ramirez, 1965 (Characidae), a freshwater teleost fish collected in the Zoological Garden of the city of Teresina (Piauí), Brazil, is described from light and transmission electron microscope observations. This myxosporean produced small plasmodia (up to 0.2 mm in diameter), each one containing all life-cycle stages of the parasite, including numerous spores. The spores, laterally compressed, averaged 54.2 (52.3-56.0) microm in total length and consisted of two unequal valves adhering together along the suture line and two caudal processes. The spore body measured 21.1 (20.0-23.1) microm in length, 5.9 (5.5-6.3) microm in width, and 2.2 (1.9-2.6) microm in thickness. The two equal ellipsoidal polar capsules of 7.4 (7.1-7.6) microm long and 1.2 (1.0-1.3) microm wide possessed a polar filament with 11-12 (rarely 13) turns. All surfaces of the spores were covered with a tightly adherent complex network of numerous densely ramified granulo-fibrillar masses, the longest measuring 1.5 microm long, observed around the caudal processes. The prevalence of infection was 30%. The taxonomic affinities of this parasite with other of the same genus in freshwater South American fish species are discussed.     

Growth and movement of secondary plasmodia of Plasmodiophora brassicae in turnip suspension-culture cells

Growth of secondary plasmodia of the clubroot pathogen Plasmodiophora brassicae was studied in dual culture of P. brassicae and turnip suspension cells. Suspension culture of P. brassicae -infected turnip cells was achieved by using P. brassicae -infected callus in Murashige and Skoog medium supplemented with 0·1 mg 2,4-D L⁻¹ and 0·02 mg kinetin L⁻¹. The shape of secondary plasmodia in suspension cells was spherical-to-subspherical. A few young plasmodia divided and became numerous spherical, small plasmodia which eventually formed a plasmodial cluster. The plasmodia fused and became vegetative plasmodia. Infected cells were significantly larger than noninfected cells. Secondary plasmodia moved within transformed turnip suspension host cells by cytoplasmic streaming of the host cells. Secondary plasmodia divided in synchrony with the transformed turnip cells.     

鱼类肝脏酯酶的同工酶构成及其对磷酸三苯酯和马拉氧磷的敏感性

用聚丙烯酰胺凝胶电泳法分离了麦穗鱼 (Pseudorasbora p arva)、金鱼 (Carassius auratus)、尼罗罗非鱼 (Tilapja nilotica)、食蚊鱼 (Gambusia affinis)、虹鳟 (Salmo gairdneri)等五种鱼的肝脏酯酶的同工酶 ,以乙酸α-萘酯为底物测定了它们的活性。在麦穗鱼、金鱼、尼罗罗非鱼体内发现两条酶带 ,在食蚊鱼和虹鳟体内发现一条酶带。离体抑制率实验表明 ,五种鱼肝脏内均含有对磷酸三苯酶敏感的酶带 ,而对马拉氧磷敏感的酶带只存在于麦穗鱼、金鱼、尼罗罗非鱼体内。酯酶同工酶分布型的意义及与马拉硫磷选择毒性的关系在本文中进行了讨论。     

Catalase activity as a potential vital biomarker of fish intoxication by the herbicide aminotriazole

The objective of this study was to investigate the effects of the herbicide 3-amino-1,2,4-triazole (AMT) on the activities of catalase and lactate dehydrogenase (LDH) in blood (plasma and erythrocytes) and eight solid tissues of goldfish, Carassius auratus. Injection of goldfish with AMT (0.5 mg/gww AMT in 0.9% NaCl) resulted in a significant decrease in catalase activity 24 h post-injection in most tissues investigated. In white and red muscle, kidney, heart, liver, brain and erythrocytes the activity of catalase decreased by 61%, 69%, 64%, 48%, 40%, 27% and 26%, respectively, in comparison to the values seen in animals injected with physiological saline (0.9% NaCl). However, the activity of LDH decreased only in red muscle (by 19%) after AMT injection, whereas in plasma it increased by 137%. Protein carbonyl levels, a measure of oxidative damage to proteins, did not change in plasma in goldfish injected with AMT and total hemoglobin levels in AMT-injected fish, although lower compared with uninjected controls, did not differ from values in saline-injected controls. It is proposed that catalase activity in erythrocytes and white muscle might be usefully developed as a potential marker for fish intoxication by aminotriazole and other related herbicides.     

Myxobolus buckei sp. n. (Myxozoa), a new pathogenic parasite from the spinal column of three cyprinid fishes from the United Kingdom

Myxobolus buckei sp. n. is described from the spinal column of Leuciscus cephalus (L.), Rutilus rutilus (L.) and Abramis brama (L.) from freshwater rivers in the North of England. The plasmodia develop within the remnants of the embryonic notochord in the intervertebral spaces. The spores are large, measuring (in microm) 14.0 +/- 0.7 x 11.5 +/- 0.6 (mean +/- SD), smooth, round to ellipsoid in valvular view with several sutural edge markings. The polar capsules are pyriform and of equal size, measuring 7.5 +/- 0.5 x 4.2 +/- 0.2 (mean +/- SD), with 11-12 turns of the polar filament arranged perpendicularly to the longitudinal axis of the polar capsule. The parasite has a large intercapsular appendix and large iodinophorous vacuole. The parasite can be differentiated from all known species of Myxobolus Bütschli, 1882 by a combination of the morphological characters defined. Infected fish show marked longitudinal compression of the body compared to uninfected individuals of the same year class, a feature which is pathognomonic for the disease. Histologically, host responses ranged from mild hypertrophy of the zygapophyseal process and expansion of the intervertebral membrane to complete hypertrophy and fusion of the vertebrae. Prominent notochord is present in the intervertebral spaces of infected fish and sporogony of the parasite leads to a vigorous focal inflammatory response involving proliferating fibroblast and osteogenic cells. The parasite causes a radial expansion of the centra and extensive dorsal and ventral outgrowths of the vertebrae leading to compression of the spinal cord and blood vessels running through the neural and haemal spines respectively. The parasite is considered highly pathogenic to juvenile cyprinids.     

Effects of temperature on the in vivo and in vitro multiplication of Trypanosoma danilewskyi Laveran et Mesnil.

Temperature directly affected the in vivo and in vitro multiplication of Trypanosoma danilewskyi Laveran et Mesnil. The parasitemia in experimentally infected goldfish (Carassius auratus L.) increased rapidly at 20 degrees C and reached its peak in 3 to 5 weeks post-infection. The multiplication was slower at 10 degrees C. The parasitemia declined after the peak at 20 degrees C. Lowering of temperature of the infected goldfish from 20 degrees C to 10 degrees C slowed multiplication of the parasite. Similarly, raising of temperature of infected fish from 10 degrees C to 20 degrees C resulted in rapid multiplication of the parasite. These were confirmed in the in vitro studies. It is suggested that lower temperature slows the multiplication of the parasites because T. danilewskyi is a parasite of a warm water host (Carassius auratus gibelio Bloch).     

Myxobolus intrachondrealis sp. n. (Myxosporea: Myxobolidae), a parasite of the gill cartilage of the common carp, Cyprinus carpio

A species not identifiable with any of the about 23 Myxobolus species recorded from the common carp so far, was detected in the gills of one- and two-summer-old specimens of the common carp ((Cyprinus carpio L.) cultured in pond farms in Hungary. The strictly tissue-specific plasmodia of the parasite were located, surrounded by hyaline cartilage cells, in the chondrous substance of the terminal parts of the gill arches and in the cartilage structure ventrally connecting the gill arches. The spores of the parasite described as Myxobolus intrachondrealis sp. n. developed in globular or ellipsoidal plasmodia measuring 300-600 microm. By their elongated ellipsoidal shape and similarly elongated polar capsules the spores were well distinguishable from the hitherto described Myxobolus species parasitic in the common carp and also from the cartilage-parasitic Myxobolus species of other fishes.     

Myxidium biliare sp. n. (Myxozoa) from gall bladder of Galaxias maculatus (Osmeriformes: Galaxiidae) in Patagonia (Argentina)

Myxidium biliare sp. n., a new myxosporean species parasitizing the gall bladder of Galaxias maculatus (Jenyns), in Patagonia, is described. Its coelozoic plasmodia were floating free in the bile. Spores are fusiform 13.7 +/- 0.9 microm long and 6.9 +/- 0.6 microm wide, with rounded ends in frontal view and slightly pointed ends in sutural view; shell with ridges and sinuous sutural line. Both maximum prevalence and maximum percentage of immature plasmodia occurred in summer. In winter the prevalence and the percentage of immature plasmodia fell to their lowest values. Prevalence was independent of host sex but increased with host length. Prevalence in 15 Patagonian Andean lakes (situated from 39 degrees 25'S to 41 degrees 30'S) ranged between 4.2% and 70%.     

A new species of the genus Gyrodactylus Nordmann, 1832 (Monogenea) from Gobio gobio (L.)

Gyrodactylus dykovae sp. n. (Monogenea: Gyrodactylidae) is described from the fins of the freshwater fish, Gobio gobio (L.) (Cyprinidae), from the Rokytná River near Moravsky Krumlov, Czechoslovakia. This species is most related to G. gobii Shulman, 1953 from which it differs by possessing more robust anchors with considerably larger shaft and by the morphology of the hook proper of marginal hooks.     

Light and scanning electron microscopy of Myxobolus porofilus sp. n. (Myxosporea: Myxobolidae) infecting the visceral cavity of Prochilodus lineatus (Pisces: Characiformes: Prochilodontidae) cultivated in Brazil

Myxobolus porofilus sp. n. is described infecting the visceral cavity of Prochilodus lineatus (Valenciennes, 1836) cultivated in S?o Paulo State, Brazil. The plasmodial form of the parasite is 3-5 mm in length and appeared compressed between the wall of the visceral cavity and the pyloric caecum, reposing on this organ. The spores are small (length 5.7 +/- 0.3 microm, width 4.8 +/- 0.2 microm; mean +/- SD) and round to elliptical in frontal view. The valve surfaces are smooth and have sutural folds. The polar capsules are ovoid, small (length 1.6 +/- 0.1 microm, width 1.1 +/- 0.1 microm) and equal in size. The polar filaments have three turns aligned perpendicularly to the longitudinal axis of the capsule. A conspicuous polar filament pore is arranged at the anterior end of the spore. The only reaction observed upon histological analysis was the presence of a capsule of connective tissue surrounding the plasmodia. This is the first report of a myxosporean parasite in the Prochilodontidae.     

Detection of Spongospora subterranea using monoclonal antibodies in ELISA

Five hybridoma cell lines secreting antibodies (MAbs) recognizing zoospores of S. subterranea were raised from splenocytes of mice. One MAb also weakly recognized plasmodia/zoosporangia and cystosori of S. subterranea , and another recognized only plasmodia/zoosporangia in plate-trapped antigen ELISA. Polymyxa graminis was recognized most strongly out of 26 micro-organisms other than S. subterranea against which the MAbs were tested. Most were recognized only weakly or not at all. The MAb that recognized zoospores of S. subterranea most strongly detected as few as three zoospores per microtitre plate well when 12 replicate wells per treatment were arranged randomly on plates and absorbance values subjected to analysis of variance. The sensitivity of detection was not improved by mixing antibodies, using a biotin-streptavidin amplification system, or by using a double antibody sandwich system. Zoospores of S. subterranea flushed from soil were detected only after unrealistically large numbers of cystosori had been added. They were not detected in samples of naturally infested soil removed from a field shortly after a severely scabbed potato crop had been harvested.     

Morphology and ultrastructure of Sphaeromyxa noblei sp. n. (Myxozoa), parasite of Heteroclinus whiteleggii (Pisces) from Australian New South Wales coast

A new species, Sphaeromyxa noblei sp. n., is described from Heteroclinus whiteleggii (Perciformes: Clinidae), a marine fish from the coast of New South Wales in Australia. This raises the number of Sphaeromyxa species to 38; their list is presented. The species is characterised by a layer of branched glycostyles, which is about 2.4 microm thick and is a feature rather unique in Myxosporea. Pansporoblasts form one or two spores. The study of ultrastructure of this species and of those described to date result in recognition of a combination of patterns characterising the genus: plasmodia have marked surface projections, the endoplasm is full of vacuoles larger than in any other myxosporean genus, and contains a special kind of cells, the lobocytes. Sections through polar capsule reveal different appearance of subsequent stretches of the polar filament unlike in other Myxosporea.     

Henneguya cynoscioni sp. n. (Myxosporea: Bivalvulida), an agent of severe cardiac lesions in the spotted seatrout, Cynoscion nebulosus (Teleostei: Sciaenidae)

A new myxosporean species, Henneguya cynoscioni sp. n., is described from the spotted seatrout, Cynoscion nebulosus (Cuvier) (Sciaenidae) as a causative agent of cardiac henneguyosis. This new myxosporean species is characterized by the morphology of spores and the sequence of SSU rDNA. Examination of 227 spotted seatrout from four South Carolina estuaries in 2008-2010 revealed a 33.5% total prevalence of H. cynoscioni. Henneguya cynoscioni produces lesions in the bulbus arteriosus, its specific site of infection. The severity of lesions and their impact on the bulbus arteriosus is proportional to the number of plasmodial stages developing in this segment of the heart, being most pronounced in host reaction directed against spores liberated from plasmodia.