Edwardsiella ictaluri invasion of IEC-6, Henle 407, fathead minnow and channel catfish enteric epithelial cells.
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Invasion of Edwardsiella ictaluri into cultured mammalian, fish and enzymatically harvested catfish enteric epithelial cells is described. Gentamicin survival assays were used to demonstrate the ability of this catfish pathogen to invade IEC-6 (origin: rat small intestinal epithelium), Henle 407 (origin: human embryonic intestinal epithelium), fathead minnow (FHM, minnow epithelial cells) and trypsin/pepsin-harvested channel catfish enteric epithelial cells. Invasion of all cell types occurred within 2 h of contact at 26 degrees C, in contrast to Escherichia coli DH5 alpha, which did not invade cells tested. Eight Edwardsiella ictaluri isolates from diseased catfish and the ATCC (American Type Culture Collection) strain were evaluated for invasion efficiency using FHM cells. All isolates were invasive, but at differing efficiencies. Invasion blocking assays using chemical blocking agents were performed on a single isolate (LA 89-9) using IEC-6 epithelial cells. Preincubation of IEC-6 cells with cytochalasin D (microfilament depolymerizer) and monodansylcadaverine (blocks receptor-mediated endocytosis) significantly reduced invasion by E. ictaluri, whereas exposure to colchicine (microtubule depolymerizer) had no effect on bacterial internalization. Results indicate that actin polymerization and receptor-mediated endocytosis are involved in uptake of E. ictaluri by IEC-6 epithelial cells. Invasion trials using freshly harvested cells from the intestine of the natural host, Ictalurus punctatus, show that invasion occurs, but at a low efficiency. This is possibly due to loss of outer membrane receptors during enzymatic cell harvest. This study provides the first documentation of the invasion of cultured mammalian and fish cells by E. ictaluri, and identifies possible mechanisms used for intracellular access. Additionally, the study describes several functional in vitro invasion models using commercially available cell lines as well as cells from the natural host (channel catfish, I. punctatus). (+info)
Site preference of myxosporean spp. on the fins of some Hungarian fish species.
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Some species of the known fish myxosporeans select fins as type location for their development. During the plasmodial development of Myxobolus, Henneguya and Thelohanellus species in cyprinid fishes and eels, 3 main locations of plasmodial development were distinguished inside the fins. M. portucalensis, M. alburni, M. caudatus and a Myxobolus sp. formed plasmodia in the dermis of the skin doublets between finrays, while plasmodia of M. diversus and H. cutanea developed inside the lumen of the cartilaginous finrays. The third type of development was represented by T. nikolskii, which started its development on the outer surface of the finrays. Plasmodia both in the lumen and on the surface of the finrays were surrounded by a cartilaginous capsule, but plasmodia of T. nikolskii also had a thick connective tissue capsule containing pieces of cartilage. (+info)
Parasitic infections in live freshwater tropical fishes imported to Korea.
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We examined 15 species of ornamental tropical fishes originating from Southeast Asia to determine the cause of losses among 8 fish farms in Korea. A total of 351 individuals belonging to 5 different families (1 species of Characidae, 6 of Cichlidae, 3 of Cyprinidae, 1 of Heleostomatidae, and 4 of Poecilidae) were collected for the purpose of detecting metazoan and protozoan parasites. Parasites were fixed and stained using routine methods, and identified. We found 3 ciliates, 2 monogeneans, 1 nematode, and 1 copepod from 7 host species. Of these, Ichthyophthirius multifiliis was the most common parasite in our study, and together with Trichodina sp., caused mass mortality of Sumatra barb Puntius tetrazona at 1 farm. We also found Camallanus cotti and Tetrahymena corlissi from guppies Poecilia reticulata, both for the first time in Korea. Farmers consider these 2 pathogens to be the most serious ones in Korea. Gussevia asota from oscar Astronotus ocellatus, and Gyrodactylus bullatarudis from platy Xiphophorus maculatus were also found in Korea for the first time. We believe that appropriate quarantine practices for tropical ornamental fishes should be introduced because the failure to require and implement quarantines has already resulted in the accidental introduction of exotic parasites to fish farms, and because these parasites can cause further economic losses if they become established in the wild. (+info)
Epizootic cutaneous papillomatosis in roach Rutilus rutilus: sex and size dependence, seasonal occurrence and between-population differences.
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Epidemiology of epidermal papillomatosis in roach Rutilus rutilus was studied in 11 roach populations in Finland in 1999 and 2000 during the spawning period. In addition, the seasonal pattern of the disease was described in 1 population. Papilloma tumours were observed on the skin and fins of roach in 8 populations out of 11. Prevalence of papillomatosis varied from 0 to 62%, being higher in populations subject to industrial or sewage effluents. Males and large fish had higher prevalence of papilloma tumours. The disease outbreak peaked during the spawning period and signs of recovered skin on fish were observed later during the year. The gender dependence in the disease prevalence found in the study may indicate differences in stress and immunocompetence between roach males and females during the spawning period. (+info)
Mutation patterns of mitochondrial H- and L-strand DNA in closely related Cyprinid fishes.
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Mitochondrial genome replication is asymmetric. Replication starts from the origin of heavy (H)-strand replication, displacing the parental H-strand as it proceeds along the molecule. The H-strand remains single stranded until light (L)-strand replication is initiated from a second origin of replication. It has been suggested that single-stranded H-strand DNA is more sensitive to mutational damage, giving rise to substitutional rate differences between the two strands and among genes in mammalian mitochondrial DNA. In this study, we analyzed sequences of the cytochrome b, ND4, ND4L, and COI genes of cyprinid fishes to investigate rates and patterns of nucleotide substitution in the mitochondrial genome. To test for strand-asymmetric mutation pressure, a likelihood-ratio test was developed and applied to the cyprinid sequences. Patterns of substitution and levels of strand-asymmetric mutation pressure were largely consistent with a mutation gradient between the H- and L-strand origins of replication. Significant strand bias was observed among rates of transitional substitution. However, biological interpretation of the direction and strength of strand asymmetry for specific classes of substitutions is problematic. The problem occurs because the rate of any single class of substitution inferred from one strand is actually a sum of rates on two strands. The validity of the likelihood-ratio test is not affected by this problem. (+info)
Induction of apoptosis in frog virus 3-infected cells.
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The ability of frog virus 3 (FV3), the type species of the family Iridoviridae, to induce apoptosis was examined by monitoring DNA cleavage, chromatin condensation, and cell-surface expression of phosphotidylserine (PS) in fathead minnow (FHM) and baby hamster kidney (BHK) cells. In productively infected FHM cells, DNA fragmentation was first noted at 6-7 h postinfection and was clearly seen by 17 h postinfection, while chromatin condensation was detected at 8.5 h postinfection. As with some other viruses, FV3-induced apoptosis did not require de novo viral gene expression as both heat-inactivated and UV-inactivated virus readily triggered DNA fragmentation in FHM cells. Moreover, FV3-induced apoptosis was blocked in FHM cells by the pan-caspase inhibitor Z-VAD-FMK, suggesting that virus infection triggers programmed cell death through activation of the caspase cascade. FV3 infection also triggered apoptosis in BHK cells as monitored by TUNEL and annexin V binding assays. To determine whether FV3, similar to other large DNA viruses, encoded proteins that block or delay apoptosis, mock- and FV3-infected FHM cells were osmotically shocked and assayed for DNA fragmentation 3 hours later. DNA fragmentation was clearly seen whether or not shocked cells were previously infected with FV3, indicating that infection with FV3 did not block apoptosis induced by osmotic shock in FHM cells. The above results demonstrate that iridoviruses triggered apoptosis and that the induction of programmed cell death did not require viral gene expression. However, it remains to be determined if virion attachment to target cells is sufficient to induce cell death, or if apoptosis is triggered directly or indirectly by one or more virion-associated proteins. (+info)
Immunohistochemistry, ultrastructure and pathology of gills of Abramis brama from Lake Mondsee, Austria, infected with Ergasilus sieboldi (Copepoda).
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Immunohistochemical, ultrastructural and pathological studies were carried out on the gills of bream Abramis brama (L.) from Lake Mondsee, Austria, that were naturally infected with Ergasilus sieboldi Nordmann, 1832. Of a total of 14 specimens of bream examined, the gills of 7 (50%) were parasitized with this copepod and the intensity of infection ranged from 1 to 23 crustaceans per host. Histopathological investigations of infected gill showed extensive tissue damage due to attachment and feeding of the crustacean. Parasites attached close to the base of filaments near the gill arch. Pressure exerted by the ectoparasite attached to the lateral margin of the gill filaments induced atrophy of the secondary lamellae. Tissue reactions included hyperplasia and mucous cell proliferation of the respiratory epithelium. Mucous cells displayed an intense immunohistochemical reactivity with the anti-nitric oxide synthase antibody. In parasitized primary and secondary lamellae, a high number of eosinophilic granular cells and rodlet cells were noticed. Rodlet cells represent an inflammatory cell type closely linked to other piscine inflammatory cells. Presence of a high number of inflammatory cells at the site of E. sieboldi attachment is related to intense host cellular reaction. (+info)
Early interactions of marine birnavirus infection in several fish cell lines.
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Marine birnavirus (MABV), a member of the genus Aquabirnavirus, family Birnaviridae, is an unenveloped icosahedral virus with two genomes of double-stranded RNA. The mechanisms of MABV adsorption and penetration are still undetermined. This work examined MABV infection in susceptible and resistant fish cell lines. MABV adsorbed not only onto the cell surfaces of susceptible (CHSE-214 and RSBK-2) cells but also onto resistant (FHM and EPC) cells. Furthermore, the virus entered the cytoplasm through the endocytotic pathway in CHSE-214, RSBK-2 and FHM cells but did not penetrate EPC cells. Thus, restriction of the MABV replication cycle is different between resistant FHM and EPC cells. The virus was found to bind to an around 250 kDa protein on CHSE-214, RSBK-2, FHM and EPC cells. Thus, this 250 kDa protein may be a major MABV receptor that exists in the plasma membranes of all four cell lines examined. This result suggests further that another receptor for virus penetration may exist in CHSE-214, RSBK-2 and FHM cells but not in EPC cells. (+info)