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1.
Abstract. A natural bath challenge method has been developed for furunculosis using Atlantic salmon, Salmo salar L., and rainbow trout, Salmo gairdneri Richardson. Fish were placed in an enclosed, continuously circulating tank system, supplemented with additional oxygen, the temperature raised gradually (overnight) to 15–16°C, a low dose of Aeromonas salmonicida (strain 184/186) introduced into the tank and the challenge maintained for 14 days. The challenge strain was characterized with respect to possible virulence factors and possessed an A-layer, ability to auto-agglutinate, haemagglutinate, adhere to Atlantic salmon cells and resist destruction by serum. No caseinase activity was detected. LD50 for salmon using this method was 1.8×103 cells per millilitre while trout had an LD50 of 9.5×104 cells per millilitre. Onset of the disease appeared to depend on fish size with larger trout (50 g) taking up to 10 days to show signs of the disease while mortalities in smaller trout (8.5g) commenced on day 1 post-challenge.  相似文献   

2.
Abstract The role of A-layer (A), protease (P) and haemolysin (H) as virulence factors of Aeromonas salmonicida, the aetiological agent of fish furunculosis, was a investigated using three strains of the bacterium. Strain MT004 lacked the A-Iayer (A−) and produced extracellular caseinase and gelatinase (P+) and haemolysin (T-lysin; H+). Strain MT028 was A−, P− and H−, and strain MT048 was A+, P+ and H−. The pathology and LD50 produced in rainbow trout by cells or extracellular products (ECP) of each strain were determined. The ECP was produced by two different methods where the protease and haemolytic activities differed in relative levels, or when the protease of MT004 ECP was inhibited by the serine protease inhibitor PMSF. The results indicate that the presence of A-layer is not essential, at least for a moderate degree of virulence; that in vitro production of extracellular proteases is not a requisite of virulent strains; that presence of protease and haemolysin in ECP can be correlated with the development of certain lesions and a rapid time to death but cannot be correlated with the lethal toxicity of the ECP. The authors conclude that an as-yet unrecognized component of ECP is responsible for killing fish.  相似文献   

3.
Abstract. Alpha foetoprotein (AFP)-like immunoreactivity was detected by double immunodiffusion in thesera of adult rainbow trout, Oncorhyachus mykiss (Walbaum) with histologically confirmed hepatocellular carcinoma experimentally induced in microinjection of embryos with aflatoxin B1 (AFB1). Partial characterization indicated that this AFP-like immunoreactivity gave immunologic cross-reaction of partial identity with human AFP. had alpha1 electrophorctic mobility, and an estimated molecular weight of 72kDa. Elevated serum AFP-like immunoreactivity levels were also measured by RIA in seven out of eight adult rainbow trout with hepatocellular carcinoma, and in three out of 17 AFB1-exposed trout without demonstrable grosws or microscopic evidence for hepatocellular neoplasms. No serum AFP-like immunoreactivity was detected or measured in normal healthy age matched DMSO-control rainbow trout. These data are consistent with the following conelusions: (1) AFP-like immunoreactivity detected and measured in the sera of rainbow trout with hepaloecllular careinoma is an analogue to human AFP; and (2) the elevated serum AFP levels measured in adult rainbow trout wilh hepatocellular careinoma resrmble those found in humans with this same malignancy. These data further suggest that serum AFP measurements might be useful to confirm the appearance of hepatocellular carcinoma in experimental fish carcinogen-assay systems, and to detect hepatocellular neoplasia in high-risk wild fish populations exposed to carcinogenie pollutants.  相似文献   

4.
Abstract. The extracellular products (ECP) of Aeromonas salmonicida , prepared by the cellophane overlay method, are lethal to rainbow trout when administered parenterally. Sublethal doses when injected i.p. or i.m. are shown to reproduce all the lesions that have been described in the literature as being associated with furunculosis. In addition, meningitis may be an important feature of furunculosi s and is reproduced by injection of ECP. A serum factor, probably an α-globulin, present in normal serum of rainbow trout, is capable of neutralizing the toxic effects of ECP. The pathology is discussed with reference to the proteolytic and leucocidal properties of the ECP and its effects upon the eosinophilic granular cells which are caused to disperse and degranulate with the possible release of histamine.  相似文献   

5.
Abstract. Controversy exists concerning the efficacy of vaccinating fish against furunculosis. Where success is claimed, there has been little attempt to characterize the protective antigens or confirm their immunogenicity. In this report, the immunogenicity of native extracellular products (ECP) of Aeromonas salmonicida and a formalin-inactivated toxoid of ECP (f-ECP) was studied in rainbow trout and rabbits, with particular attention to the putative bacterial virulence factors protease and haemolysin. Using crossed immunoelectrophoresis and Protein-A absorption, antibodies to seven ECP components were detected in the rabbit following immunization with native ECP; antihaemolysin antibodies were found but antibodies to the protease could not be detected. Antibodies to at least 14 components of ECP, including haemotysin and protease, were detected in the rabbit following immunization with f-ECP. In trout immunized either with native ECP or f-ECP, antibodies to only four ECP antigens were detected and no antibodies to haemolysin or protease were found. The results may explain previous reports that passive immunization with rabbit antisera gave superior protection against furunculosis compared with antisera raised in fish, and indicate that many extracellular antigens of A. salmonicida may require modification in order to improve their immunogenicity in fish.  相似文献   

6.
Abstract. An enzootic, Australian, atypical strain of Aeromonas salmonicida isolated from diseased goldfish, Carassius auratus (L.), was inoculated into Atlantic salmon, Salmo salar L., brown trout, S. trutta L., rainbow trout, S. gairdneri Richardson, and brook trout, Salvelinus fontinalis (Mitchill), fingerlings by intraperitoneal injection (i.p.) and by bath challenge, the latter with and without prior abrasion of skin. The 10-day LD50 (i.p.) was estimated to be 7·4 × 10-3 colony forming units (cfu) for Atlantic salmon, 3·0 × 10-2 cfu for brown trout, 3·7 × 102 cfu for brook trout and 6·4 × 103 cfu for rainbow trout. Brown, rainbow and brook trout succumbed to bath challenges with between 105–106 cfu/ml, developing ulcers of the skin and septicaemia. The organism was trasmitted from inoculated fish to five of 195 within-tank control fish via water and established a carrier state in one of 14 Atlantic salmon. It was concluded that the organism poses a significant threat to the salmonid farming industry and wild salmonid fisheries in Australia.  相似文献   

7.
Abstract. The pathogenicity of Renibacterium salmoninarum to carp, Cyprinus carpio L., and rainbow trout, Salmo gairdneri Richardson, was investigated. All carp injected with 4·8 × 108 cells/fish, or 4·8 × 107 cells/fish survived for 38 days. R. salmoninarum was isolated from all moribund fish, but not from the kidney of surviving fish, although R. salmoninarum antigen was detected in several of these fish by the dot blot assay. On the other hand, mortality in rainbow trout was 95% in the fish injected with 4·8 × 108 cells/fish, and 15% in those which received 4·8 × 107 cells/fish. R. salmoninarum antigen was detected by the dot blot assay in all surviving rainbow trout. The number of R. salmoninarum cells was immediately decreased by carp or rainbow trout serum, and the serum bactericidal activity of carp was higher than that of rainbow trout. Carp blood leucocytes had higher phagocytic activity than those of rainbow trout.  相似文献   

8.
Mycobacterium spp. isolated from food and ornamental fish in Thailand (TB1, TB40, TB267 and TB268), and the type strains Mycobacterium marinum (NCIMB 1298), M. fortuitum (NCIMB 1294) and M. chelonae (NCIMB 1474) were cultured in Long's medium, Eagle's minimum essential medium, Sauton's medium and modified Sauton's medium. The latter enabled excellent growth and production of extracellular products (ECP) from TB 40, TB267, TB268 and M. marinum NCIMB 1298 in particular, whereas growth and production of ECP for all strains was limited in Long's medium. SDS-PAGE protein profiles of ECPs from 14-day culture supernatants showed major bands at 65 and <14 kDa. After 2 days culture at the higher temperature of 37°C (heat shock), the production of ECP from all mycobacteria strains except M. marinum averaged approximately four- to 10-fold higher than from strains cultured for 14 days at 28°C. Enzyme testing for the type strains indicated only mucinase activity for M. marinum, while lipase and RNase activities were detected for M. chelonae and M. fortuitum . Protease and DNase activities could not be detected for any of the Mycobacterium spp. tested. The medium lethal dose (LD50) of ECP to rainbow trout and Nile tilapia was greater than 400 μg protein fish–1.  相似文献   

9.
Abstract. The specificity of the major protease secreted by Aeromonas salmonicida has been explored using a number of proteins and p-nitroanilides as substrates. The 70kDa protease was found to hydrolyse two p-nitroanilides which have been reported to be specific substrates for thrombin. Kinetic parameters (kcat, and Km) were compared for the 70kDa protease and for thrombin as were the effects of a number of inhibitors. The 70kDa protease is able to degrade proteins which have a relatively open structure, for example, caseins or denatured bovine scrum albumin, to small fragments mostly of Mr<2500. However, proteins with a more compact structure are more resistant to the protease. It was concluded that the 70kDa protease shows some of the specificity features of thrombin, although it is less discriminating in its choice of both low and high Mr substrates than thrombin. In preliminary experiments, the 70 kDa protease was found, like thrombin, to decrease the clotting time of rainbow trout blood. The possible physiological significance of these results is discussed.  相似文献   

10.
ABSTRACT:   Thermal tolerance was studied in a rainbow trout strain successively selected through high temperature breeding at 20–27°C since 1966 in Miyazaki Prefecture, Japan. The hatching rate and fry mortality at high temperatures were examined in the selected strain along with normal strains cultured at water temperature of 9–17°C. The hatching rate of embryos fertilized at either 10 or 14°C and subsequently subjected to high temperatures in the blastula or neurula stage of the selected strain, was marginally higher than that of the normal strain counterparts. The upper 50% lethal temperatures (LT50) for embryos in the early segmentation, blastula and neurula stages of the selected strain were also higher than those of the normal strain counterparts. Death temperatures and LT50 of fries acclimated to 20°C of the selected strain were significantly higher than those of the normal strains. However, no difference in the critical thermal maximum was detected between the different strains. These results suggest that the selected strain of rainbow trout established by selecting successively for many generations at high temperatures acquired a degree of thermal tolerance.  相似文献   

11.
Haemorrhagic areas in the mouth of farmed turbot, Scophthalmus maximus (L.)   总被引：4，自引：0，他引：4  
Abstract. An epizootic in juvenile turbol reared on a farm located in The Ria de Vigo, Galicia, Northwest Spain, is described. The diseased turbot did not display unusual swimming behavior; the external signs of the disease were abdominal distension and haemorrbagic areas in the mouth. Internal examination of the fist showed an accumulation of reddish fluid in the peritoneal cavity. Microbiological analysis of the diseased fish revealed the presence, in pure culture in all the organs and lesions examined, of a bacterium which was characterized biochemically as Vibrio splendidus biotype I. The virulence tests showed that the V. splendidus biotype I isolate was pathogenic for rainbow trout (LD50: 2.2 × 104) and also for turbot (LD50: 1.2 × 104). The treatment of the fish using flumequine incorporated into the feed was effective in offsetting the mortality rate.  相似文献   

12.
The present study evaluates if abdominal dis-tension caused by a water-filled stomach in seawater farmed rainbow trout, Oncorhynchus mykiss (Walbaum), and Atlantic salmon, Salmo salar L., could be provoked experimentally by live chilling in sea water (0.5 °C). Fifty rainbow trout and 50 salmon were visually classified, either as normal or suffering from the condition. Prior to chilling, no rainbow trout or salmon suffered from a water-filled stomach. During chilling, 25% of the rainbow trout and 2.5% of the salmon developed water-filled stomach. Affected rainbow trout had significantly higher blood plasma osmolality and significantly lower body weight than normal trout. The frequency of trout with a water-filled stomach increased significantly with increasing chilling time and increasing plasma osmolality. The regression coefficients revealed that the proportion of affected rainbow trout in-creased by 0.82% h−1 of chilling and by 0.46% per unit increase in mosmol L−1. The present study revealed that abdominal distension in seawater farmed rainbow trout was far more severe than in Atlantic salmon and could be provoked experimentally by osmoregulatory stress. This is consistent with the observation that water-filled stomach appear only occasionally in seawater farmed Atlantic salmon.  相似文献   

13.
Abstract. Intravascular injection of purified 70-kDa protease or GCAT-LPS from Aeromonas salmonicida , or both, invariably led to consumptive coagulopathy within 2h in Atlantic salmon, Salmo salar L. By entering the coagulation cascade at two different levels, the two enzymes work in concert in thrombus formation, the significance being that circulatory failure is probably the major cause of death in acute furunculosis. Only intravascular injection of GCAT-LPS led to consumptive coagulopathy in rainbow trout, Oncorhynchus mykiss (Walbaum), despite the fact that trout received only half the GCAT dose, and/or four times the protease activity administered to salmon. These results indicate that salmon is, by far, the most susceptible to protease and suggest that rainbow trout is the most susceptible to GCAT-LPS. The substrate profile of the purified protease gave supporting evidence that it works as activated coagulation factor X. The protease is inhibited in vitro by antithrombin and by α2-macroglobulin and both inhibitors are consumed in vivo in response to intravascular administration of the enzymes, thus showing a potential for AT and α2M to inhibit the protease also in vivo. Provided such plasma antiprotease activities are correlated with resistance to the disease, and the inhibitors show genetic variation they would be promising candidates for indirect selection and hence a means to prevent furunculosis independently of vaccines. Circulating inhibitors of GCAT-LPS remain to be identified.  相似文献   

14.
Abstract. Both fingerling and adult rainbow trout were given intraperitoneal inoculations of the 131P2 reovirus which did not result in mortalities but produced a subclinical disease in 56% of the adults and in 65% of the juvenile fish. Sera from virus infected adult trout contained virus neutralizing and precipitating antibodies and demonstrated immunofluorescence of antigen(s) specific for the 13P2 virus. Virus infection was characterized by increased virus titres and a multifocal reticulo-endothelial granulomatous hepatitis, occasionally accompanied by a fibrosing pancreatitis. Electron microscopy of infected liver tissues demonstrated arrays of 13P2 virus particles in a reticuloendothelial cell phagosome and in a hepatocyte lipid inclusion body from one of three juvenile fish examined. These results show that 13P2 virus inoculation of rainbow trout causes a non-virulent virus infection accompanied by specific humoral antibodies. These antibodies are sufficient in titre for use in future serologic studies of the virus. Further discussion regarding the pathogenesis of this virus in bluegills versus rainbow trout is presented.  相似文献   

15.
The effect of hydrogen peroxide (H2O2) treatment on the clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout was examined. In untreated control fish, clearance was characterized by a rapid initial phase, with 50% reduction in bacterial antigen in the first 12 h after exposure. The bacteria then cleared more gradually, with total clearance being achieved by 40 h. Treatment of fish with 100 p.p.m. H2O2 did not influence bacterial clearance compared to untreated controls. Exposure for 1 h to H2O2 concentrations ≤ 100 p.p.m. caused no detectable clinical signs or evidence of ultrastructural damage to the respiratory epithelium. However, levels in excess of this caused significant gill damage.  相似文献   

16.
Abstract. Aeromonas salmonicida was studied at fish farms producing salmonid smolts in northern and central Finland from 1982 onwards. Aeromonas salmonicida subsp. salmonicida has been causing epizootics in salmon, Salmo salar L., and trout, S. trutta L., at two coastal farms in northern Finland sinee 1986, involving 1–29% mortality in the fish-rearing units affceted. The disease causes more serious losses of sea trout yearlings and brood fish than of salmon. The achromogenic atypical A. salmonicida proved to be the most common bacterial disease in brown and sea trout at one farm in northern Finland throughout the period, causing constant heavy losses, mainly of fingerlings, especially in 1982–1986. It was found only occasionally in central Finland. Pigment-producing atypical A- salmonicida caused mortality among brown trout at one farm in northern Finland and one in central Finland.  相似文献   

17.
Abstract. Infection trials using two serotypes of VHS viruses (type 1 and 23/75) demonstrated that Atlantic salmon fry were susceptible to the disease when injected intraperitoneally (i.p.) with 103 pfu of virus/fish but resistant to infection by a bath method when exposed for 3 h in water containing 5 × 104 pfu of virus/ml. In the i.p.-infected fish, mortality reached 78 and 67% within 13 days with VHSV1 nad 23/75 serotypes, respectively. High virus yields were recovered from infected fish and virus shedding was demonstrated by the onset of VHS in rainbow trout kept in the outflow water from the aquaria containing infected salmon. Neither mortality nor virus shedding occurred in salmon infected by the water route but virus multiplication was demonstrated in 2 of 60 fish with VHSV1 and 3 of 60 fish with virus 23/75. On day 79 post-infection the sera from surviving salmon of both i.p. and bath infection trials exhibited good neutralizing titres (around 1000) against the homologous viruses.  相似文献   

18.
Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L−1, or for 10 min at 50 mg L−1, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L−1), buffered MS-222 (100 mg L−1) with NaHCO3 (100 mg L−1), and unbuffered MS-222 (100 mg L−1) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO3 treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.  相似文献   

19.
A selection of 16 field isolates of Photobacterium damselae from marine rainbow trout farms in Denmark was subjected to phenotypic and genotypic characterization and pathogenicity to fish. All isolates belonged to the subspecies damselae , being positive for haemolysis, motility and urease. There were considerable differences in haemolytic properties, some isolates presenting a broad zone of haemolysis and others only a narrow zone. Pulsed-field gel electrophoresis revealed a high diversity indicating that P. damselae subsp. damselae is an opportunistic, not clonal pathogen in Danish marine rainbow trout. Virulence of the strains to rainbow trout was highly variable with LD50 values ranging from 3.9 × 103 to 1.5 × 108 cfu at 20 °C. The virulence was significantly higher at 20 °C than at 13 °C. The strains with the strongest haemolytic properties were the most virulent suggesting a strong involvement of haemolysin in the pathogenesis. The pathological changes were consistent with a bacterial septicaemia and the haemorrhages were more pronounced than for most other bacterial infections.  相似文献   

20.
Abstract. Viral haemorrhagic septicaemia (VHS) is reported for the first time in sea-water cultured rainbow trout. Heavy mortalities with typical signs and lesions A VHS virus (serotype 1) was isolated from the diseased fish. The mortalities were caused only by the VHS virus and 80 days post transfer of trout to sea-water the mortalities reached 85%, of the initial population.
The disease was experimentally transmitted to rainbow trout, both in sea-water 3·104 pfu/ml of virus or by intramuscular injection of various doses of VHS1 (7·101 7·104 or 7·104 pfu per fish). Death occurred in all infected groups and started earlier in sea-water. Typical signs of VHS were observed in moribund fish. Viral multiplication was demonstrated to have occurred in fish organs.  相似文献   

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