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1.
Abstract. An enzyme-linked immunosorbent assay using dried blood on filter paper, was developed for the detection of antibodies against the haemoflagellate Cryptobia salmositica in juvenile rainbow trout, Oncorhynchus mykiss (Walbaum). Each fish (average weight about 5g) in three experimental groups was either inoculated with 20000 attenuated live C. salmositica vaccine, or inoculated with 2000 or 20000 pathogenic parasites per fish. The vaccine was effective in protecting juvenile trout 4 weeks after vaccination and antibody titers were higher in vaccinated and challenged fish than in unvaccinated and infected ones. Specific antibodies were detected one week post-infection (w.p.i.) with the pathogen and declined to low levels at 6 w.p.i. The high-dose group (20000 per fish) had antibody titres comparable to those of the vaccinated and challenged fish. 相似文献
2.
Adult rainbow trout, Oncorhynchus mykiss (Walbaum), maintained in either fresh or sea water were vaccinated with a live Cryptobia salmositica vaccine. All vaccinated fish were protected 4 weeks later against the cryptobiosis, while unvaccinated rainbow trout developed the disease (e.g. high parasitaemia and severe anaemia) after challenge with virulent C. salmositica . There was also no disease in vaccinated fish when they were transferred from fresh to sea water immediately after vaccination. Complement fixing antibodies (CFAbs) were detected in vaccinated fish and the CFAbs lysed parasites under in vitro conditions. The antibody titres increased rapidly at one week post-challenge in vaccinated fish in fresh water and vaccinated fish transferred from fresh water to sea water after vaccination. However, the production of CFAbs was delayed by one week in vaccinated fish in sea water and the antibody titre was significantly lower than that in fish maintained in fresh water. 相似文献
3.
Hatchery-reared Atlantic salmon, Salmo salar L., were vaccinated intraperitoneally (i.p.) with a live attenuated Cryptobia salmositica vaccine (either 100 000 or 5000 parasites fish−1 ) and 4 weeks later were challenged with the parasite (either 100 000 or 5000 parasites fish−1 ). Unvaccinated, infected salmon had high parasitaemias and were anaemic. Fish given a high dose (100 000 parasites fish−1 ) had higher parasitaemias than fish given the lower dose. Vaccinated fish had low parasitaemias and a mild anaemia, but recovered quickly after challenge. Complement-fixing antibody increased in vaccinated fish after challenge and was highest at 2 weeks post-challenge. The cell-mediated response (both T cells and B cells) was depressed in infected fish until 4 weeks after infection. In vaccinated fish, the humoral response (i.e. B-lymphocytes) was greater than the cell-mediated response (i.e. T-lymphocytes). In contrast, infected fish had a greater cell-mediated than humoral immune response. 相似文献
4.
Abstract. Rainbow trout that recovered from experimental Cryptobia salmositica infection 6 and 10 weeks earlier were protected against multiple intraperitoneal challenges of 50 000 and 10 000 parasites isolated from infected fish. The immunity was non-sterile; low parasitaemias were detected following a larger challenge (112 000 parasites). The indirect haemagglutination test was used to detect C. salmositica -specific agglutimns. Antibody titers increased during the first 18 weeks of infection. The infectivity of cultured C. salmositica was neutralized by incubation in heat-inactivated immune plasma. Infectivity of C, salmositica from infected fish was not neutralized by similar treatment. Complement fixing antibody was detected using the in vitro immune lysis test. Immune lysis occurred when cultured C. salmositica were used. Adoptive transfer of both leucocytes and plasma from immune fish conferred partial protection against the parasite in naive recipients. Complement fixing antibody may be important during early acute infection while phagocytosis may be important during the later chronic phase. 相似文献
5.
Abstract— Due to species introductions, brook charr (Salvelinus fontinalis) and rainbow trout (Oncorhynchus mykiss) occur together in many North American streams and typically exhibit a pattern of distribution in which brook charr numerically dominate headwaters and rainbow trout dominate downstream reaches. It has been suggested that 1) the two species compete or 2) the two species do not compete because they are differentially adapted to environmental conditions found in upstream and downstream zones. We assessed whether there were differences in growth and macrohabitat (pool, run and riffle) selection of brook charr and rainbow trout in upper, middle and lower stream zones of a small Pennsylvania stream. Brook charr and rainbow trout placed in replicate paired enclosures set in upstream and downstream reaches showed no significant differences in growth and survival rates upstream, but brook charr had significantly greater growth rates than rainbow trout downstream. Enclosed fish and free-ranging fish both had negative growth rates during the summer. Enclosed fish lost significantly less weight than free-ranging fish. Instantaneous growth rates of free-ranging adult brook charr and rainbow trout from May to August were negative for both species in all stream zones. Underwater observations of adult brook charr and rainbow trout showed both species occurred significantly more often in pool macrohabitats than expected on the basis of macrohabitat availability, except for rainbow trout in the upstream zone. The proportion of pool macrohabitat was not significantly different among stream zones. Brook charr do not appear to be better adapted to upstream environments in Powdermill Run based on growth, survival and macrohabitat selection during summer. Negative biotic interactions acting along with differential environmental adaptations may explain the pattern of distribution of brook charr and rainbow trout in streams, but long-term transplant experiments with additional life stages will be necessary to examine this hypothesis. 相似文献
6.
Abstract. An experimental Cryptobia salmositica infection in rainbow trout, Salmo gairdneri Richardson, produced suppression of the humoral response against sheep red blood cells as measured by direct haemagglutination. Two-month and 5-month infections produced equal suppression. The parasite also produced suppression of the humoral response against a bacterial pathogen, Yersinia ruckeri . Anti- Y. ritckeri titres were significantly lower in most fish infected with C. salmositica than in non-infected fish. Immunosuppression became evident when C. salmositica first appeared in the blood (first 2 weeks of infection), Immunosuppression was confirmed by challenge with Y. ruckeri . Mortality at challenge occurred in 64·3% to 83·3% of the fish already infected with C. salmositica at the time of initial Y. ruckeri exposure. There was no mortality at challenge if fish were not infected with C. salmositica at initial bacterial exposure, nor in those concurrently infected with both pathogens. Antigenic competition may have caused the immunosuppression. 相似文献
7.
Abstract. Intraperitoneal implants of cortisol (cortisol suspended in hydrogenated coconut oil) were used to induce a graded hypercortisolism in rainbow trout, Salmo gairdneri Richardson. There was no obvious reduction in circulating lymphocytes in cortisol-implanted rainbow trout (70, 140 or 210μg/g body weight). Cortisol-implanted fish infected with Cryptobia salmositica had significantly higher parasitaemia and lower antibody litres compared with controls infected with haemonagellate but given coconut oil implants. These confirm the immunodepressive effects of the steroid. The parasite was also more readily detected at the early stage of the infection (shorter prepatent period, more infected fish and higher parasitaemia) in cortisol-implanted fish (140 and 210 μg/g body weight) than in controls. The mortality of the infected cortisol-implanted fish was higher than that of the infected fish implanted with only coconut oil, or the cortisol-implanted but non-infected fish. This in vivo study suggests that protective immunity against C. salmositica is, in part, due to a humoral response. 相似文献
8.
Abstract. There are two basic antigenic components in Cryptobia that are responsible for the anaemia in infected rainbow trout. A 'lytic component', which is dosage-dependent, causes lysis of red blood cells independent of antibody or complement. The second, an 'immune-complex-forming component', attaches to red blood cells, forms immune complexes with specific antibody and activates complement resulting in haemolysis. These two antigenic components, from both live and lysed Cryptobia , were present in the serum of infected fish. When sonicated antigen or heat-inactivated antiserum (from infected fish) was incubated with red cells from uninfected fish, a portion of the red cells was lysed and a positive Coombs' reaction was observed with the remaining intact red cells. The positive Coombs' reaction was due to immune complexes adsorbed onto the red cells and these lysed when incubated with complement. Antibody by itself did not adsorb onto the red cells. From the fourth week post-infection, a positive Coombs' reaction was observed in all infected fish and haemolysis occurred with complement. The authors suggest that, in infected fish, one or more components of the complement cascade is depleted continually during infection and that the anaemia is due to the lytic action of the antigen and immune complex formation on red cells. These lead to intra-vascular haemolysis as well as erythrophagocytosis. In general, the mechanism of anaemia in cryptobiosis appears similar to that in African trypanosomiasis. 相似文献
9.
A monoclonal antibody (designated MAb-007) was produced against the pathogenic haemoflagellate Cryptobia salmositica Katz. This IgG3 antibody recognized the 47-kDa antigenic polypeptide of C. salmositica (SDS-PAGE and Western immuno-blotting). The antibody did not agglutinate live parasites, and there was no change in the staining intensity of the 47-kDa band on Western immunoblots after immunoabsorption of MAb-007 with live intact parasites. The 47-kDa antigen recognized by MAb-007 was localized in the cytoplasm of the parasite (immunogold labelling and electron microscopy). The monoclonal antibody cross- reacted with the 47-kDa polypeptides of C. bullocki Srrout and C. catostomi Bower & Woo. It was used in an antigen-capture ELISA for the detection of parasite antigen in the plasma of rainbow trout inoculated with the parasite, or with an attenuated vaccine strain of C. salmositica. All pre-infection plasma were negative while all infected fish with detectable parasitaemias were positive for antigen at 1–9 weeks after infection. Parasite antigen was even detected in vaccinated fish that were negative for parasites using the wet mount microscopic technique. The antigen-capture ELISA detected C, salmositica antigen in whole cell lysate preparations at concentrations as low as 0.5 μg ml-1. Fifty microlitres of fish plasma was required in the antigen-capture ELISA, and the use of a plate reader and 96-well plates facilitated rapid analysis of a large number of plasma samples. The sensitivity of the assay makes it a potentially useful tool for detection of Cryptobia infections. 相似文献
10.
Abstract. Delayed-type hypersensitivity (DTH) reaction (an in vivo manifestation of cell-mediated immunity) was detected in Oncorhynchus mykiss maintained on a pantothenic-acid-supplemented diet 2 weeks after infection with Cryptobia salmositica. The reaction was similar to that in mammals with mononuclear cell infiltration into the dermis and muscle layers and the presence of oedema. DTH reaction was also displayed by fish on a pantothcnic-acid-supplemented diet that had recovered from the infection and were protected against further infection. The reaction was less marked in infected or protected fish on a pantothenie-acid-deficient diet. Inhibition of macrophage migration (an in vitro expression of cell-mediated immunity) was observed when head kidney cell suspensions from protected fish maintained on either pantothenic acid supplemented or deficient diets were incubated with Cryptobia antigen. No inhibition of migration was evident when head kidney cell suspensions from the above fish were incubated without antigen, nor was it evident when cells from uninfected fish were used. The occurrence of a typical DTH reaction in rainbow trout and the feasibility of assessing it by measuring the thickness of the induration provides a simple and practical method for assessing cell-mediated immunity in large scale vaccination programmes against pathogens. 相似文献
11.
Woo PT 《Journal of fish diseases》2003,26(11-12):627-646
Salmonid cryptobiosis is caused by Cryptobia (Trypanoplasma) salmositica. The haemoflagellate has been reported from all species of Pacific Oncorhynchus spp. on the west coast of North America. It is normally transmitted by the freshwater leech, Piscicola salmositica, in streams and rivers, and sculpins, Cottus spp., are considered important reservoir hosts. The pathogen can also survive on the body surface of fish because it has a contractile vacuole to osmoregulate when the fish is in fresh water. This allows for direct transmission between fish, especially in aquaculture facilities. The parasite divides rapidly by binary fission in the blood to cause disease, the severity of which is directly related to parasitaemia. Cryptobia salmositica has a mitochondrium and it normally undergoes aerobic respiration; however, if its mitochondrium is damaged it will switch to glycolysis. Its glycolytic enzymes and catalase are contained in glycosomes. Cysteine protease is a metabolic enzyme, and its neutralization inhibits oxygen consumption and multiplication of the parasite. An important virulent factor in cryptobiosis is a secretory metalloprotease. The protective mechanism involves production of complement fixing antibodies, phagocytosis by macrophages, and cell-mediated cytotoxicity. Recovered fish are protected, probably for life as the immunity is non-sterile. Clinical signs of the disease include anaemia, anorexia, splenomegaly, general oedema and abdominal distension with ascites. The metabolism and swimming performance of infected fish are significantly reduced and the bioenergetic cost of the disease is very considerable. Fish are susceptible to hypoxia and their immune system is depressed during acute cryptobiosis. Severity of the disease and mortality rates vary significantly between species and stocks of salmon. Protective strategies include selective breeding of Cryptobia-resistant fish. This is innate resistance to infection and it is controlled by a dominant Mendelian locus. In these fish the parasite is lysed via the alternative pathway of complement activation. In Cryptobia-tolerant fish (infected with the pathogen but which do not suffer from disease) the metalloprotease secreted by the parasite is neutralized by alpha2 macroglobulin. Hence, the production of a transgenic Cryptobia-tolerant salmon is an option. This strategy has the advantage in that human intervention (e.g. vaccination, chemotherapy) is not required once the transgenic fish is produced. Acquired immunity is another option; a single dose of the attenuated live vaccine protects fish for at least 2 years. The protective mechanism in vaccinated fish is similar to that in recovered fish. The trypanocidal drug, isometamidium chloride, is an effective therapeutic and prophylactic agent. It accumulates in the mitochondrium of the parasite and significantly disrupts aerobic respiration by causing lesions in the organelle. Efficacy of the drug is significantly increased after its conjugation to antibodies. This immuno-chemotherapeutic strategy has the advantage in that it will lower the drug dosage and hence side-effects of chemotherapy. It will probably reduce the accumulation of the drug in fish, an important consideration in food fish. 相似文献
12.
Rabbit anti-thymocyte serum (RATS) against thymocytes of rainbow trout was toxic to leucocytes from intact and thymectomized rainbow trout at 10 °C under in vitro conditions. The total number of leucocytes decreased significantly in 24 h after RATS was injected intraperitoneally into intact rainbow trout, but the number returned to pre-injection level within 1 week. RATS destroyed a lower percentage of leucocytes in thymectomized fish than in intact fish under both in vitro and in vivo conditions and the recovery in the number of leucocytes was slower in thymectomized fish. The parasitaemia, packed cell volume and production of complement fixing antibody in thymectomized and intact fish (injected with RATS before Cryptobia salmositica infection) were not significantly different from control fish (not injected with RATS), and they both acquired protective immunity against cryptobiosis on recovery. This indicates that RATS is not cytotoxic to B-like cells in the lymphoid tissue which produce complement fixing antibody against C. salmositica and that the protective antigen in C. salmositica seems to be thymus-independent. 相似文献
13.
A monoclonal antibody (IgG 1) (designated as MAb-001) was produced against the pathogenic haemoflagellate Cryptobia salmontica Katz. The antibody agglutinated live parasites under in vitro conditions. Live C. salmositica, incubated with MAb-001 at 10 °C, did not multiply and were dead within 4 weeks in culture. About 50% of juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), inoculated intraperhoneally with C. salmositica, incubated in MAb-001 prior to inoculation, did not become infected, while in adult rainbow trout, the peak parasitaemia was reduced. These results indicate that MAb-001 is a protective monoclonal antibody and the antigen it recognizes is located on rhe surface membrane of C. salmositica. The antibody also inhibits multiplication and affects viability of the parasite under in vitro conditions. 相似文献
14.
Abstract. Oxygen consumption of juvenile rainbow trout (5 g at 13°C) at moderate swimming speeds did not change significantly when infected with Cryptobia salmositica. However, significant reductions of as much as 44% of the maximum aerobic scope for activity and 24% of the critical swimming speed were observed when the parasitaemia reached a maximum of 57.6 × 106 ml−1 fish blood at 3 weeks post- infection. Blood haematocrit was significantly reduced from the initial 34.1 to 19.7% at 4 weeks post- infection, probably as a result of haemolysis by the parasite. The destruction of red blood cells clearly led to lower oxygen carrying capacity, and reduced respiratory and swimming performance. 相似文献
15.
Triploid hybrids between female rainbow trout Oncorhynchus mykiss and male brook charr Salvelinus fontinalis, Arctic charr S. alpinus and lake charr S. namaycush, together with diploid and triploid rainbow trout controls from the same dams, were tested in freshwater farming up to their fourth year of life. All hybrids displayed lower survival rates than the controls, the weakest genotype being the Arctic charr hybrid. Mortalities were mostly observed at the embryonic and larval stages and at the adult stage as a consequence of male sexual maturation. Growth of all hybrids was hindered (compared with controls) during the first year, but only moderate differences remained after 3 years. Sexual maturation resulted in a weight inferiority of males in all genotypes. As to carcass traits, female hybrids displayed a slightly higher dressing percentage than female triploid rainbow trout, as a result of lower visceral losses. These results are discussed with reference to hybrid resistance to rhabdoviruses from the viewpoint of fish farming improvement. 相似文献
16.
Rowena Hoare Sung‐Ju Jung Thao P. H. Ngo Kerry L. Bartie Kim D. Thompson Alexandra Adams 《Journal of fish diseases》2019,42(2):229-236
Flavobacterium psychrophilum is one of the most important pathogens affecting cultured rainbow trout (Oncorhynchus mykiss). Recent information from UK salmonid farms showed country‐wide distribution of genetically and serologically divergent clones, which has hampered the development of a vaccine for rainbow trout fry syndrome. The current study assessed the efficacy of an injectable polyvalent vaccine containing formalin‐inactivated F. psychrophilum in rainbow trout. The vaccine was formulated with an oil adjuvant (Montanide ISA 760VG) or formalin‐killed cells alone. Duplicate groups of trout (60 ± 13 g) were given phosphate‐buffered saline or vaccine formulated with Montanide by intra‐peritoneal (i.p.) injection and challenged by intra‐muscular (i.m.) injection with a homologous and a heterologous isolate of F. psychrophilum at 525 degree days post‐vaccination (dd pv). Significant protection was achieved in vaccinated fish (p = 0.0001, RPS 76% homologous, 88% heterologous). Efficacy of the adjuvanted vaccine was also demonstrated by heterologous challenge at 1155 dd pv resulting in 100% protection, whereas survival in the un‐adjuvanted group was not significantly different from control fish. Levels of specific antibody at 1155 dd pv, as measured by ELISA, were significantly higher in the fish vaccinated with adjuvant when compared with unvaccinated fish. 相似文献
17.
The trypanocidal drug isometamidium chloride (Samorin) was conjugated to polyclonal and monoclonal antibodies produced against the pathogenic haemoflagellate Cryptobia salmositica . Under in vitro conditions the unconjugated drug normally accumulates rapidly in the kinetoplast in the parasite; however, once it was conjugated to antibodies (either polyclonal or monoclonal) it was found throughout the parasite. Isometamidium conjugated to polyclonal antibodies lysed C. salmositica under in vitro conditions, but parasites were not agglutinated. In contrast, isometamidium conjugated to monoclonal antibodies (against a 200 kDa surface membrane glycoprotein) did not lyse C. salmositica , but parasites were agglutinated. Because of the low efficacy of the monoclonal conjugate against the parasite in vitro , its cryptobiocidal effect was not evaluated further. The infectivity of C. salmositica (incubated either in culture medium or whole blood) was reduced in fish after in vitro exposure to isometamidium conjugated to polyclonal antibodies. Parasitaemias were reduced in infected chinook salmon, Oncorhynchus tshawytscha, after treatment with isometamidium conjugated to polyclonal antibodies. 相似文献
18.
L. C. LLEWELLYN 《Journal of fish diseases》1980,3(1):29-39
Abstract. Blood spots in the eye and haemorrhages in the musculature of the caudal peduncle were the main indication of disease in Australian hatchery reared brook trout Salvelinus fontinalis (Mitchill) and Atlantic salmon Salmo salar L. Mortalities of 0·1-0·5% per day occurred for 3–4 weeks in fish stocked in circular ponds in the months of February and March when temperatures were falling from 23–11·C. The bacterium belonging to the Family Enterobacteriaceae had similarities with Yersinia ruckeri (the RM bacterium) and Serratia liquefaciens . As a temporary designation SBS was used. It is recommended that its taxonomic position remain sub judice for the present. Inoculation experiments on brook trout and landlocked Atlantic salmon indicated that signs appeared 2–7 days after inoculation and that rainbow trout Salmo gairdneri Richardson were resistant to the disease. 相似文献
19.
Susceptibility and antibody production against pathogenic and vaccine strains of the haemoflagellate, Cryptobia salmositica were investigated in five full‐sib families (A–E) of Atlantic salmon, Salmo salar. Humoral response and susceptibility of families were compared within three treatments: infection, vaccination and vaccination followed by challenge. Parasitaemias caused by the vaccine strain of C. salmositica were considerably lower than those caused by the pathogenic strain. All vaccinated families were protected when challenged with the pathogenic strain. Family B had significantly lower parasitaemias (with both strains) than the other families. When naïve fish were infected with the pathogenic strain, this family had a significantly lower and earlier peak parasitaemia (4.3 ±1.3 × 106 parasites mL?1 blood at 3 weeks post‐infection; w.p.i.) than the other families. Family C had the highest peak (11.1 ± 1.2 × 106 parasites mL?1 blood), which occurred at 4 w.p.i. Antibodies against C. salmositica were detected earlier in Family B (3 w.p.i.) than in Family C (5 w.p.i.). This demonstrates an association of increased susceptibility with a delayed antibody response. Western immunoblot identified antibodies against 112, 181 and 200 kDa antigens earlier in more resistant fish (Family B). Antigenic stimulation leading to a stronger antibody response was shown with the vaccine strain and in the later stages of infection. 相似文献
20.
J M Blanc Frédéric Vallée Patrick Maunas & Jean-Paul Fouriot 《Aquaculture Research》2005,36(2):120-129
Related sib‐groups of rainbow trout × brown trout and rainbow trout × brook charr triploid hybrids and monospecific diploid and triploid rainbow trout controls were obtained from a common set of rainbow trout dams. On the basis of hybrid juvenile performances, 10 sib‐groups were selected and the corresponding diploid rainbow trouts were raised up to adult stage. Females from each group of rainbow trout were used to produce a second generation of hybrid progeny, the performances of which were analysed for grandmaternal variation and relation with first‐generation relatives. Results showed that hybrid traits (alevin yield and weight, survival and growth of fingerlings) were strongly influenced by maternal origin, and could be correlated to those of rainbow trout controls, but that maternal abilities had a low rate of inheritance. It was concluded that little improvement can be expected through selective breeding within parental populations. 相似文献