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1.
Extranuclear histones and histone fragments (13–15 kDa) in gill microsomes of a phylogenetic series of teleostean fishes (tilapia, trout, carp, eel and elephant fish) were studied. Histones H2A and H2B are the first occurring of these antimicrobial agents. Halfway the evolution of teleosts (carp, trout) H3 fragments emerge, which disappear in later stages of development and are replaced by truncated H1 (tilapia). The hypothesis is forwarded that variation in composition of extranuclear histones is caused by changes in specificity of histone transporters.  相似文献   

2.
The ability of rainbow trout, Oncorhynchus mykiss (RBT), to produce a localized mucosal immune response was investigated following intraperitoneal (i.p.) or peranal (p.a.) immunization with a protein-hapten carrier, fluorescein isothiocyanate conjugated to keyhole limpet haemocyanin (FITC/KLH). Antibody levels in serum, mucus, tissue culture supernatant from blood and spleen leucocytes, and excised skin, intestine and gill tissues were determined by ELISA. Significantly, elevated antigen-specific antibodies were elicited in both serum and mucus of fish immunized i.p. Mucosal antibody responses, in general, paralleled serum responses over time. Leucocytes isolated from spleen and blood of i.p. immunized fish at week 10 produced significantly elevated antibody levels against FITC when cultured in vitro. Excised skin, intestine and gill tissues from these fish also exhibited significantly elevated antibody responses indicating localized production in the mucosa from tissue-specific B cells. A localized mucosal immune response was elicited only after i.p. and not p.a. immunization, suggesting that systemically stimulated B cells migrate to mucosal tissues where they produce antibodies locally.  相似文献   

3.
During a 3‐month period from June to the end of August 2016, ~5% mortalities were observed in a farm with rainbow trout (Oncorhynchus mykiss Walbaum) and one farm of common carp (Cyprinus carpio L.) in Bulgaria. The disease was manifested by gill ulcers/rot, asphyxiation and bloody ascites. Aeromonas hydrophila was isolated from the internal organs of all the diseased fish. Bacillus mycoides or B. pseudomycoides were recovered from the gill lesions on diseased carp and rainbow trout, respectively, with identification achieved by conventional phenotyping and by sequencing of the 16S rRNA gene. In vivo experiments confirmed that all three organisms were pathogenic to rainbow trout.  相似文献   

4.
The objective of this study was to determine whether exposure of rainbow trout (Oncorhynchus mykiss) to water containing a stressed trout or skin extract from stressed and non-stressed trout would elicit a stress response in conspecifics. Juvenile rainbow trout were exposed for 1 hour to water containing a stressed fish, homogenized skin extracts from a non-stressed fish, skin extract from a stressed fish and water with none of these factors. The stress response was measured over a 24-h period (1, 6, 12, 24 h after exposure). Plasma cortisol levels increased at 12 h in fish exposed to water from a stressed fish and skin extract from a stressed fish. Plasma glucose and hepatic hsp70 levels were not affected by treatments. The results suggest that rainbow trout elicit a stress response when exposed to stress-related alarm cues released from conspecifics.  相似文献   

5.
Abstract.— The effect of temperature on production and affinity of antibodies against antigens from the parasitic ciliate Ichthyophthirius multifiliis were studied in rainbow trout ( Oncorhynchus mykiss ). Fish were immunized with I. multifiliis antigens and reared at three different temperatures, 5, 12, and 20 C for 56 d. The production of specific antibodies was examined by enzyme-linked immunosorbent assay (ELISA). The results showed that temperature has a pronounced effect on the assay result of the ELISA technique. Plasma samples analyzed at 5 C showed that the specific antibody response of fish reared at 5 C was similar to fish reared at 12 and 20 C. However, when samples were assayed at 12 and 20 C, the measured antibody response tended to be higher for the samples from trout reared at 12 and 20 C. Additionally, it was found that rainbow trout reared at 5 C showed a delayed but not hampered antibody response compared to fish reared at 12 and 20 C, indicating a correlation between low temperature and delayed antibody production.  相似文献   

6.
Glugea plecoglossi (Microspora) is a significant cause of economic loss in cultured ayu, Plecoglossus altivelis, in Japan, due to the unsightly appearance of infected fish harbouring xenomas in the body cavity. Modes of transmission of G. plecoglossi via the skin and digestive tract were studied in an experimental infection model using rainbow trout, Oncorhynchus mykiss. Combined with Uvitex 2B and in situ hybridization (ISH) assays, the early development of G. plecoglossi was successfully traced. Following a bath exposure of fish Uvitex 2B-labelled G. plecoglossi spores were observed to attach to microscopic injuries (trypan blue-positive sites) of fish skin, after which ISH-positive sporoplasms were found to invade the epidermis as early as 5 min post-infection (PI), migrating rapidly to the subdermis. It was also shown that G. plecoglossi entering via the skin does not spread into the internal organs but develops into subdermal xenomas. After rainbow trout were exposed to G. plecoglossi spores by oral intubation, spores germinated in the intestinal lumen, followed by penetration of sporoplasms into the gut mucosal epithelium 5 min PI. In vitro trials determining stimulation factors (fish mucus, changes in pH, digestive enzymes) for the extrusion of the polar tube were inconclusive. The present study indicates that skin wounds and the gut epithelium can be portals of entry of G. plecoglossi and that natural infection in fish seems to occur perorally rather than via the skin.  相似文献   

7.
Serum and mucosal antibody responses of juvenile rainbow trout, Oncorhynchus mykiss, were characterized by enzyme‐linked immunosorbent assay (ELISA) following immunization with various preparations of formalin‐killed Flavobacterium psychrophilum cells. The protective nature of these preparations was then determined by immunizing rainbow trout fry and challenging with the bacterium. Juvenile rainbow trout immunized intraperitoneally (i.p.) with formalin‐killed F. psychrophilum emulsified with Freund's complete adjuvant (FCA), and i.p. with formalin‐killed F. psychrophilum either with or without culture supernatant generated significant serum antibody responses by 6 and 9 weeks, respectively. Significant mucosal antibody responses were detected by 9 weeks only in fish immunized i.p. with killed F. psychrophilum/FCA. Following immunization and bacterial challenge of rainbow trout fry, protective immunity was conferred in F. psychrophilum/FCA and saline/FCA groups with relative per cent survival values of up to 83 and 51, respectively. Significant protection was not observed in treatment groups immunized by immersion or i.p. without adjuvant at the challenge doses tested. Results suggest that stimulation of non‐specific immune factors enhances the ability of fish to mount a protective immune response, but specific antibody appears necessary to provide near complete protection. In this study, an ELISA was developed to monitor anti‐F. psychrophilum antibody production in trout. The relationship of such responses to protective immunity suggests that future vaccination strategies against coldwater disease may require stimulation of both the innate and adaptive arms of the immune response.  相似文献   

8.
Efficacy of mineral oil-based experimental injection vaccines against Flavobacterium psychrophilum were tested in rainbow trout, Oncorhynchus mykiss (Walbaum), under laboratory and field conditions. The vaccines consisted of formalin- or heat-inactivated whole bacterium cell preparations of two different serotypes (Fd and Th) or a combination of serologically different F. psychrophilum (Fd and/or Th and/or Fp(T);Th). Specific antibody responses against the bacterium in plasma and skin mucus were evaluated post-vaccination with enzyme-linked immunosorbent assay. Efficacy of the vaccinations was determined by challenge trials to F. psychrophilum with the vaccinated rainbow trout. Significantly higher antibody levels in plasma were detected in vaccinated fish compared with mock-vaccinated fish. Injection vaccination did not trigger specific antibody production in the skin mucus. Significantly higher survival of i.p. vaccinated fish compared with non-vaccinated fish was observed during the challenge. The results suggest that mineral oil-based injectable vaccines containing formalin- or heat-inactivated virulent cells of F. psychrophilum effectively triggered specific antibody production and protected the fish against bacterial cold water disease.  相似文献   

9.
Japanese people consume crop bran, which contains relatively high amounts of gamma-oryzanol (ORZ), as foodstuffs and food materials. We have recently confirmed that ORZ inhibits NF-κB activation, activates peroxisome proliferator-activated receptor gamma (PPARγ), and increases plasma adiponectin levels. ORZ is therefore expected to improve lipid and carbohydrate metabolisms and various inflammatory diseases, i.e., metabolic syndrome including type 2 diabetes. ORZ administration also allowed fish, such as rainbow trout, yellowtail, and red sea bream, to accumulate protein through enhancement of lipid and carbohydrate metabolisms. In the present study, we have investigated ORZ accumulation levels in mouse and rainbow trout administered ORZ-containing feed. Although mouse muscle and liver hardly contained ORZ, muscle tissues of every fish species accumulated higher amounts of ORZ. These findings suggest that rainbow trout, red sea bream, and yellowtail accumulate ORZ in muscle.  相似文献   

10.
Abstract. The degree of protection in immunized rainbow trout against the infective larval stages of Diplostomum spathaceum was investigated. Immunized rainbow trout demonstrated a 70–80% reduction in the number of metacercariae establishing in their eye-lenses following challenge with live cercariae compared with unimmunized fish. Both normal trout serum and anti–diplostomule serum weTC cytotoxic to diplostomules in vitro . However, there was no correlation between the degree of protection and the level of serum antibody within individual fish. Reduced establishment of migrating diplostomules, indicative of in vivo killing, occurred in fish possessing activated macrophages and challenged with cercariae opsonized with specific antibody. An antibody–dependent cell–mediated cytotoxic (ADCC) mechanism is proposed, conferring protection in immunized fish. This is supported by the finding that cercarial and diplostomule antigens are capable of activating macrophages in immunized rainbow trout. The role of macrophage activation and specific antibody in conferring protection in immunized fish is discussed.  相似文献   

11.
A feeding experiment was conducted to evaluate the supplemental effect of bile salts to a defatted soybean meal-based non-fish meal diet for rainbow trout Oncorhynchus mykiss. A fish meal-based diet (FM) and two non-fish meal diets with and without bovine bile salts (SC+B and SC, respectively) were fed to fish (13 g initial weight) for 10 weeks. Fish fed diet SC showed inferior growth and feed efficiency, while bile salt supplementation improved the parameters to the same levels as fish fed diet FM. Crude fat and starch digestibility of diet SC-fed fish decreased after the 10-week feeding trial compared to the data obtained with fish that had no experience of the diet. Total biliary bile salt content and intestinal maltase activity of fish fed diet SC were the lowest among treatments, while these parameters were improved by bile salt supplementation. Morphological changes occurred in the distal intestine and liver of the diet SC group, although the histological features of fish fed diet SC+B were similar to those of fish fed diet FM. These results suggest that bile salt supplementation to a soybean meal-based diet improves the nutrient utilization by normalizing digestive processes in rainbow trout.  相似文献   

12.
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   

13.
Three experiments were conducted in order to evaluate the effects of urea and trimethylamine oxide (TMAO) supplementation in diets for seawater‐farmed rainbow trout Oncorhynchus mykiss. In all experiments, commercial extruded feed was used as basal control diet, whereas experimental diets were the basal diet supplemented with either 10 g kg–1 TMAO, 10 g kg–1 urea or 10 g kg–1 TMAO and 10 g kg–1 urea. The experiments were conducted both in research units and in a commercial fish farm. Quality was evaluated in three different commercial slaughterhouses, sorting the rainbow trout into superior, ordinary or processing grade. Significantly more TMAO was observed in muscle of rainbow trout given dietary supplementation of TMAO. However, the levels were low compared with levels in marine fish in general. Trimethylamine (TMA) was not found. No increase in muscle urea content was observed in trout fed urea in the diet. Significantly reduced fat belching, seen as reduced number of fat droplets surfacing in the net pen, was observed in groups given TMAO‐supplemented feeds. At least twice as many rainbow trout with skin vesicles were observed in the control group as in the group fed urea‐supplemented diet. In addition, skin vesicles developed into open sores in the winter only in rainbow trout fed control diet. In a large‐scale trial, quality grading of almost 200 000 market‐sized rainbow trout showed significantly more (88 vs. 83%) trout of superior grade in the group fed dietary urea compared with fish fed control diet. The main reason for downgrading from superior to ordinary and processing grade was skin lesions, supporting the observations made in the experimental units.  相似文献   

14.
Abstract. Laboratory-raised Cryptobia -susceptible brook charr, Salvelinus fontinalis (Mitchill), and rainbow trout, Oncorhynchus mykiss (Walbaum), were vaccinated intraperitoneally with a live Cryptobia salmositica vaccine (250000 parasites per fish), and 4 weeks later were challenged with the pathogen (250000 parasites per fish). Unvaccinated and infected brook charr had high parasitaemias but no clinical signs of disease, while unvaccinated and infected rainbow trout had anaemia and general oedema. Vaccinated and challenged fish had very low parasitaemias compared to unvaccinated and infected brook charr and rainbow trout. Complement fixing antibodies were detected in vaccinated and challenged fish 2 weeks after challenge. Unvaccinated and infected brook charr had consistently higher litres of complement fixing antibody than unvaccinated and infected rainbow trout. Parasitaemias were lower in all fish in which titres of complement fixing antibody were high. In a second experiment, brook charr inoculated intraperitoneally or intramuscularly with 100000 C. salmositica per fish had high parasitaemias but no anaemia or other clinical signs. The results show that susceptible brook charr do not suffer from cryptobiosis and may serve as reservoir hosts for C. salmositica in areas where the disease is prevalent. Vaccination to reduce the parasitaemia when fish become infected may be a control strategy in these areas.  相似文献   

15.
The trout fishery at Draycote Water was investigated during the 1980 fishing season. Fish stocked were batch-marked according to the date of introduction by freeze branding, and catch data were obtained by the cooperation of anglers. Population estimates were made at the end of the fishing season using gill nets and mark-recapture techniques. Of the 32,960 marked brown and rainbow trout stocked, 69.8% were caught and declared by anglers. Returns of rainbow trout were better (78.1%) than those of brown trout (44.2%). Over 90% of all fish caught were taken within 45 days of stocking. Catch per unit effort fluctuated widely but was closely associated with stockings of fish. Catchability (Q) of stocked fish was found to diminish rapidly with time after stocking. At the end of the fishing season the estimated population of marked trout was 4045 (2 × SE = 250), compared with a theoretical number (stock less total catch) of 10,802, giving a mean daily apparent natural mortality, made up of the true natural mortality plus the undeclared catch, of 1.36%.  相似文献   

16.
Preliminary studies were conducted to determine if several feed supplements with the potential to improve dietary mineral availabilities in fish meal had any measurable effect in fish feeds. In the first study with rainbow trout, 11 supplements were tested: citric acid; sodium citrate; potassium chloride; sodium chloride; histamine dihydrochloride; EDTA disodium salt; sodium bicarbonate; a mixture of amino acids; ascorbic acid; a mixture of inositol and choline; and cholecalciferol. Apparent availability of calcium, phosphorus, magnesium, sodium, iron, manganese and strontium in fish meal-based diets was determined using both yttrium oxide (Y2O3) and chromium oxide (Cr2O3) as inert dietary markers. Apparent availability was expressed as the fractional net absorption (%) of minerals from diets. After a 7-day acclimation period with test diets, fecal samples were collected for five consecutive days using passive collection systems. Apparent availability of calcium, phosphorus, magnesium, iron, manganese and strontium was increased by citric acid supplementation. Apparent availability of manganese also was increased by EDTA and sodium citrate. The other supplements had no measurable effect on the apparent availability of minerals in fish meal. In the second study, the effect of supplemental citric acid was further investigated using monogastric (rainbow trout) and agastric fish (goldfish). Fish were fed for 5 weeks (rainbow trout) or 3 weeks (goldfish) with fish meal-based diets containing either 0% (control), 2% or 5% citric acid on a dry basis. Feces were collected by settling and by stripping. Apparent availabilities of calcium and phosphorus were greatly affected by citric acid supplementation in rainbow trout but not in goldfish. Phosphorus levels in feces of fish fed a diet with 5% citric acid were approximately half of that of fish fed the control diet (0% citric acid) in the rainbow trout trial. This pattern was consistent during the 5-week feeding trial. A dietary supplement of citric acid as high as 5% did not reduce feed intake or appetite of rainbow trout. Conversely, this level of dietary acidification led to a marked reduction of feed intake in goldfish. Dietary supplementation of citric acid at 2% level did not reduce feed intake of goldfish; however, this level of dietary acidification had little effect on the apparent availability of major minerals in fish meal-based diet. Levels of non-fecal excretion of calcium and phosphorus, inorganic phosphorus in urine, and citric acid in feces were increased in rainbow trout fed 5% citric acid. The pH values of the feces and urine were decreased in rainbow trout fed citric acid. Plasma bicarbonate, plasma calcium and phosphorus, and blood pH of rainbow trout tended to increase by a 5% dietary supplementation of citric acid. The soluble inorganic phosphorus content increased in the diets and decreased in the feces of rainbow trout by supplementing the diet with 5% citric acid. Feces samples of rainbow trout collected by stripping provided similar availability values to data collected by settling for most elements except sodium, which had negative values in all dietary treatments.  相似文献   

17.
Abstract. The epidemiology of epizootic haematopoietic necrosis virus (EHNV) infection was studied in farmed rainbow trout, Oncorhynchus mykiss (Walbaum). Estimates of mortality during five outbreaks on a commercial farm from 1986 to 1992 ranged from 0033 to 0.2% per day and total mortality did not exceed 3–4% in any outbreak. Affected fish were 0+ and less than 125 mm forklength. Clinical signs were non-specific, and laboratory examination was required to confirm the diagnosis. At the height of the outbreak in 1992, EHNV was demonstrated by virus isolation and antigen capture ELISA in 89% of clinically affected fish and 51% of dead fish, while the prevalence of infection in apparently healthy in-contact fish was 4%. Two and 4 months later the virus was not detected in a group of apparently healthy fish that had been affected earlier. Antibodies specific for EHNV were not found in rainbow trout from the infected farm; however, strong humoral responses were detected by ELISA in two immunized fish, indicating that the virus was immunogenic. These data suggested that EHNV was poorly infective but highly virulent in rainbow trout. Clinical EHNV infection was positively correlated with high rearing density and a low rate of water exchange, and therefore, with presumed poor water quality. Water temperature, which ranged from 11 to 17°C during outbreaks, did not appear to determine the incidence of clinical infection. EHNV infection in farmed rainbow trout was preceded by infection in free-living redfin perch, Perca fluviatilis L., in the water catchment, but it was uncertain whether this represented the source of infection for rainbow trout.  相似文献   

18.
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   

19.
Heat shock proteins (Hsps) can promote integrity of damaged intestinal epithelial tissues. The aim of this study was to determine the effects of a commercially available anti‐phospholipase A2 antibody on growth, feed efficiency, Hsp expression, and intestinal health of rainbow trout fed diets containing high levels of soybean meal (SBM). Six practical‐type diets (45% protein; 15% fat) were formulated to contain 0, 17.5, or 35% SBM with and without antibody. Each diet was fed to three replicate tanks of fish (20 fish/tank; initial weight 21 g) for 9 wk. RNA was isolated from liver and intestinal tissues (n = 3/tank) to detect alterations in Hsp 27, Hsp 70, and Hsp 90 expression. Fish fed a diet containing 35% SBM had decreased growth and intestinal damage. Hsp 27 and Hsp 90 mRNA was elevated in fish fed 35% SBM. Fish fed 35% SBM with antibody had lower Hsp 90 mRNA expression than fish without antibody. These data indicate that feeding rainbow trout high levels of SBM alters the expression of some Hsps and anti‐phospholipase A2 antibody supplementation does not normalize growth or prevent distal enteritis in fish fed 35% SBM.  相似文献   

20.
Viral haemorrhagic septicaemia (VHS) is one of the most important viral diseases in rainbow trout that has caused great losses to Iranian rainbow trout aquaculture industry in the last 3 years. Therefore, rapid and reliable diagnosis of VHS virus infections is of great importance. An enzyme linked immunosorbent assay (ELISA) method was performed to study serum antibodies against viral haemorrhagic septicaemia virus (VHSV) using recombinant fragments of their N protein. For this purpose, the virus was first isolated from an infected farm. A part of the nucleocapsid (1–505 bp) gene was amplified by RT‐PCR using specific primers. The amplified fragment was ligated to pMALc2x vector and transferred to DH5α strain of Escherichia coli. Then, recombinant plasmids were tested for protein expression in E. coli Rosetta strain. SDS‐PAGE analysis indicated the production of a recombinant protein with an expected molecular weight of 61 KDa. Analysis of trout serum samples from seven previously infected farms and two VHS free farms showed that the designed ELISA method was effective in diagnosing the infected fish. The results revealed that the developed serological assay using designed ELISA based on recombinant protein (N) has the potential to be used in monitoring studies and to determine the prevalence of VHS in rainbow trout farms. The present data allow evaluating the levels of nonneutralizing antibodies without crude virus preparations.  相似文献   

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