Gaffkemia in the California spiny lobster, Panulirus interruptus: Infection and immunization

Infection (gaffkemia) can be induced in the California spiny lobster (P. interruptus) with injections of Pediococcus homari (formerly Gaffkya homari). An LD_{5 0} of 10³ microbes per ml hemolymph is observed at 17°C. Prior immunization of lobsters (P. interruptus) with an avirulent strain of P. homari protects lobsters against 100 × LD_{5 0}. This is the first report of acquired immunity to P. homari in a susceptible species. The high incidence of gaffkemia in American lobsters (Homarus americanus) held in live holding facilities makes prophylactic measures essential in regions outside the enzootic area of the North Atlantic to avoid inadvertent transplantation of the disease to other susceptible populations of animals.     

Aeromonas salmonicida infectivity studies in goldsinny wrasse, Ctenolabrus rupestris (L.)

Goldsinny wrasse, Ctenolabrus rupestris (L.). were experimentally and naturally infected with Aeromonas salmonicida. Goldsinny wrasse were found to be susceptible to infection with A. salmonicida but significantly more resistant to infection than Atlantic salmon, Salmo salar L. The pathology of the acute infection is described. Following infection significantly higher levels of agglutinating antibody were detected in the sera of recovered wrasse than those observed in the Atlantic salmon controls. However, a large proportion of the recovered wrasse were culture positive for A. salmonicida and appeared to have entered a chronic stage of infection quite distinct from the carrier status that can develop in Atlantic salmon. This study indicates that, although goldsinny wrasse are susceptible to A. salmonicida, these fish are unlikely to become primarily infected, but contract furunculosis from infected Atlantic salmon. However, goldsinny wrasse may act as a reservoir for subsequent infections of cultivated Atlantic salmon because of the development of a chronic form of the disease. The potential for the vaccination of goldsinny wrasse against furunculosis is discussed.     

Development of a Piscirickettsia salmonis immersion challenge model to investigate the comparative susceptibility of three salmon species

Piscirickettsia salmonis, the aetiological agent of salmonid rickettsial septicaemia (SRS), is a global pathogen of wild and cultured marine salmonids. Here, we describe the development and application of a reproducible, standardized immersion challenge model to induce clinical SRS in juvenile pink (Oncorhynchus gorbuscha), Atlantic (Salmo salar) and sockeye salmon (O. nerka). Following a 1‐hr immersion in 10⁵ colony‐forming units/ml, cumulative mortality in Atlantic salmon was 63.2% while mortality in sockeye salmon was 10%. Prevalence and levels of the bacterium in kidney prior to onset of mortality were lower in sockeye compared with Atlantic or pink salmon. The timing and magnitude of bacterial shedding were estimated from water samples collected during the exposure trials. Shedding was estimated to be 82‐fold higher in Atlantic salmon as compared to sockeye salmon and peaked in the Atlantic salmon trial at 36 d post‐immersion. These data suggest sockeye salmon are less susceptible to P. salmonis than Atlantic or pink salmon. Finally, skin lesions were observed on infected fish during all trials, often in the absence of detectable infection in kidney. As a result, we hypothesize that skin is the primary point of entry for P. salmonis during the immersion challenge.     

Outbreak of viral haemorrhagic septicaemia (VHS) in lumpfish (Cyclopterus lumpus) in Iceland caused by VHS virus genotype IV

A novel viral haemorrhagic septicaemia virus (VHSV) of genotype IV was isolated from wild lumpfish (Cyclopterus lumpus), brought to a land‐based farm in Iceland, to serve as broodfish. Two groups of lumpfish juveniles, kept in tanks in the same facility, got infected. The virus isolated was identified as VHSV by ELISA and real‐time RT‐PCR. Phylogenetic analysis, based on the glycoprotein (G) gene sequences, may indicate a novel subgroup of VHSV genotype IV. In controlled laboratory exposure studies with this new isolate, there was 3% survival in the I.P. injection challenged group while there was 90% survival in the immersion group. VHSV was not re‐isolated from fish challenged by immersion. In a cohabitation trial, lumpfish infected I.P. (shedders) were placed in tanks with naïve lumpfish as well as naïve Atlantic salmon (Salmo salar L.). 10% of the lumpfish shedders and 43%–50% of the cohabiting lumpfish survived after 4 weeks. 80%–92% of the Atlantic salmon survived, but no viral RNA was detected by real‐time RT‐PCR nor VHSV was isolated from Atlantic salmon. This is the first isolation of a notifiable virus in Iceland and the first report of VHSV of genotype IV in European waters.     

Genetic variation in susceptibility of Atlantic salmon, Salmo salar L., to furunculosis, BKD and cold water vibriosis

Genetic variation in susceptibility of Atlantic salmon, Salmo salar L., to furunculosis, bacterial kidney disease (BKD) and cold water vibriosis was studied by challenge testing one-year-old fingerlings. Fish from 81 full-sib families within 32 sire progeny groups were infected with Aeromonas salmonicida, Renibacterium salmoninarum and Vibrio salmonicida. Estimated heritabilities were relatively low, being highest for BKD (h²= 0.23) and lowest for cold water vibriosis (h²= 0.13). Genetic correlations between the ability to survive the diseases were all positive, but the magnitude of the genetic correlation between furunculosis and BKD may be biased upwards because some of the dead BKD fish were also infected with furunculosis. The application of selection to develop resistant populations of Atlantic salmon is advocated. Challenge testing seems to be a feasible method, with relatively low costs and easy management. The future response to selection will depend on the relationships between results from a challenge test and mortalities under farming conditions and between disease resistance and other traits in the breeding goal.     

脂肪和蛋白质水平对工业养殖大西洋鲑消化酶、非特异性免疫及水质的影响

为探讨脂肪和蛋白质水平对工业化养殖大西洋鲑相关消化酶活力、免疫及对水质指标的影响,采用3×2双因素试验设计(3脂肪水平:18%、21%、24%,即F18、F21、F24;2蛋白质水平:38%、48%,即P38、P48),形成6种实验处理的膨化配合颗粒料,每处理3重复,在工业化封闭循环海水养殖(RAS)条件下,选用初重(650.0±45.50)g大西洋鲑720尾进行56 d的养殖实验。结果显示:(1)F21可显著提高大西洋鲑胃、肠、肝脂肪酶活力,比低脂肪分别提高11.52%、14.63%、4.31%;P48使肠、肝胰蛋白酶活力分别提高8.23%、8.33%,且发现其活力水平远高于胃蛋白酶。(2)F18、P48可显著提高肠道AKP酶活力,F18比F21、F24分别提高18.61%、31.70%,P48比P38提高13.69%。(3)F21、F24可显著改善血清的抗氧化能力,F21比F18的SOD活力提高10.32%,同时MDA含量降低4.49%;P48有利于提高血清LZM活力及补体C3含量,P48比P38分别高9.49%、5.93%。(4)低蛋白质水平可显著降低水体中氨氮、硝酸盐的含量,P38比P48分别降低61.70%、28.36%;提高脂肪水平可降低水体氨氮含量,F21、F24比F18分别降低10.00%、8.20%。研究表明,饲料脂肪和蛋白质水平与消化吸收酶、非特异免疫、养殖水氨氮之间关系特征明显;适当提高脂肪水平有利于提高消化道脂肪酶活力,提高蛋白质水平有利于提高免疫力;低蛋白质和中高脂肪组合可有效降低大西洋鲑的氨氮排泄量。     

Saprolegnia diclina IIIA and S. parasitica employ different infection strategies when colonizing eggs of Atlantic salmon,Salmo salar L.

Here, we address the morphological changes of eyed eggs of Atlantic salmon, Salmo salar L. infected with Saprolegnia from a commercial hatchery and after experimental infection. Eyed eggs infected with Saprolegnia spp. from 10 Atlantic salmon females were obtained. Egg pathology was investigated by light and scanning electron microscopy. Eggs from six of ten females were infected with S. parasitica, and two females had infections with S. diclina clade IIIA; two Saprolegnia isolates remained unidentified. Light microscopy showed S. diclina infection resulted in the chorion in some areas being completely destroyed, whereas eggs infected with S. parasitica had an apparently intact chorion with hyphae growing within or beneath the chorion. The same contrasting pathology was found in experimentally infected eggs. Scanning electron microscopy revealed that S. parasitica grew on the egg surface and hyphae were found penetrating the chorion of the egg, and re‐emerging on the surface away from the infection site. The two Saprolegnia species employ different infection strategies when colonizing salmon eggs. Saprolegnia diclina infection results in chorion destruction, while S. parasitica penetrates intact chorion. We discuss the possibility these infection mechanisms representing a necrotrophic (S. diclina) vs. a facultative biotrophic strategy (S. parasitica).     

Mycobacterium salmoniphilum infection in farmed Atlantic salmon, Salmo salar L

Multiple greyish‐white visceral nodules containing abundant rapidly growing and acid‐fast bacteria, subsequently identified as Mycobacterium salmoniphilum, were detected in moribund and newly dead market‐sized fish during a period of increased mortality in an Atlantic salmon, Salmo salar, farm in western Norway. Isolates cultured from diseased fish were phenotypically consistent with Mycobacterium sp. previously isolated from Atlantic salmon [MT 1890 (= NCIMB13533), MT1892, MT1900 and MT1901] in the Shetland Isles, Scotland. Partial sequences of 16S rDNA, ribosomal RNA internal transcribed spacer (ITS1), 65‐kDa heat‐shock protein (Hsp65) and β subunit of RNA polymerase (rpoB) revealed 97‐99% similarity with M. salmoniphilum type strain ATCC 13758^T. The source of infection was not confirmed. Koch’s postulates were fulfilled following experimental challenge of Atlantic salmon with field isolate NVI6598 ( FJ616988 ). Mortality was recorded in experimentally infected fish; however, the infection remained subclinical in the majority of affected fish over the 131‐day challenge period.     

Comparison of NADH-dependent cytochrome b5 reductase activity and in vitro methemoglobin induction by sodium nitrite in Oncorhynchus mykiss, Salmo salar, and Salvelinus fontinalis

Methemoglobin is hemoglobin containing ferric iron. Methemoglobin cannot bind to oxygen and at high concentrations causes tissue hypoxia. Brook trout (Salvelinus fontinalis) develop significantly greater methemoglobinemia than Atlantic salmon (Salmo salar) or rainbow trout (Oncorhynchus mykiss) following general anesthesia with benzocaine or tricaine methanesulfonate. The objective of this study was to compare the activity of the major methemoglobin reducing enzyme, NADH-dependent cytochrome b5 reductase (CB5R), in brook trout erythrocytes to the activity of CB5R in Atlantic salmon and rainbow trout erythrocytes. Methemoglobin levels were compared using co-oximetry following in vitro incubation of erythrocytes with sodium nitrite (NaNO₂). The CB5R activity was measured using a ferricyanide assay. There was significantly greater methemoglobin at time 0 in brook trout erythrocytes than in rainbow trout or Atlantic salmon erythrocytes (2.79 ± 0.29 %, 2.19 ± 0.23 %, 2.08 ± 0.14 %), (P < 0.001). There was significantly greater methemoglobin induction by NaNO₂ in brook trout erythrocytes (33.14 ± 3.32 %) than in rainbow trout or Atlantic salmon erythrocytes (28.73 ± 2.92 % and 24.85 ± 1.40 %, respectively), (P < 0.001). The CB5R activity was significantly less in brook trout erythrocytes (median of 3.05 μmol/min/μl) than in rainbow trout erythrocytes (median of 6.73 μmol/min/μl). The CB5R activity in Atlantic salmon erythrocytes (median 4.09 μmol/min/μl) was not significantly different than in brook or rainbow trout erythrocytes. Total methemoglobin at any one time is a balance between induction by oxidants and reduction by antioxidants. Lower CB5R activity in brook trout erythrocytes may contribute to a species-specific sensitivity to methemoglobin induction; however, there are likely additional factors.     

Humoral immune response of Japanese eel, Anguilla japonica Temminck & Schlegel, to Pleistophora anguillarum Hoshina, 1951 (Microspora)

A humoral response of Japanese eel, Anguilla japonica Temminck & Schlegel, to the microsporean Pleistophora anguillarum Hoshina was demonstrated using immunoblotting and an enzyme-linked immunosorbent assay (ELISA). Japanese eel immunoglobulin was purified by affinity chromatography. The immunoglobulin was composed of 25-kDa light chains and 72-kDa heavy chains. The ELISA values of P. anguillarum antibodies in naturally infected fish sera were significantly higher than those of clinically healthy fish. Spore proteins from the microsporean were separated by electrophoresis and subjected to analysis by Western blot. Sera from naturally infected fish showed different reaction patterns to the spore proteins. While the sera randomly selected from naturally infected eels all showed a significant positive reaction to P. anguillarum antigens, the mucus from only three out of the nine infected eels reacted positively in the ELISA test. Subsequent analyses indicated that there was no significant difference in the amount of mucus immunoglobulin among the tested eels. Therefore, the generally lower ELISA values of mucosal anti-P. anguillarum antibodies from the infected eels tested were evidently not caused by a lack of immunoglobulin per se, but seem to be the result of a lack of anti-P. anguillarum antibodies in the mucus and/or a lower affinity in the anti-P. anguillarum antibodies that were present.     

Inactivated Piscine orthoreovirus vaccine protects against heart and skeletal muscle inflammation in Atlantic salmon

Heart‐ and skeletal muscle inflammation (HSMI) caused by infection with Piscine orthoreovirus (PRV) is one of the most common viral diseases in farmed Atlantic salmon (Salmo salar) in Norway, and disease outbreaks have been reported in most countries with large‐scale Atlantic salmon aquaculture. Currently there is no vaccine available for protection against HSMI, partly due to the lack of a cell line for efficient virus propagation. Erythrocytes are the primary target cells for PRV in vivo and a potential source for isolation of PRV particles. In this study, PRV was purified from infected erythrocytes, inactivated and used in a vaccination trial against HSMI. A single immunization with adjuvanted, inactivated PRV induced protection against HSMI in Atlantic salmon infected by virus injection 6 weeks later, while a moderate protection was obtained in fish infected through natural transmission, i.e. cohabitation. The PRV vaccine significantly reduced PRV loads and histopathological lesions typical for HSMI compared to the unvaccinated control group. This is the first demonstration of protective vaccination against PRV, and promising for future control of HSMI in Atlantic salmon aquaculture.     

The effect of domestication on a brown trout (Salmo trutta m fario) broodstock in Hungary

Molecular markers (PCR–RFLP and microsatellite) were used to investigate the genetic background of the only brown trout (Salmo trutta m fario) broodstock in Hungary which due to the hydrogeography of the country should theoretically belong to the Danubian lineage. PCR–RFLP (mitochondrial DNA control region and lactate dehydrogenase and somatolactin genes) as well as microsatellite (BFRO002, OMM1064, Ssa408uos, SsoSL417, SsoSL438) markers were used to distinguish between Danubian and Atlantic lineages of brown trout in the Lillafüred broodstock. Altogether 435 fish were tagged during the experimental period. Due to mortalities, fin clips were collected from 401 individuals. According to the genetic analysis of the mitochondrial DNA, the Danubian haplotype is present in only one individual (0.2 %) of the broodstock. Analysis of the nuclear markers revealed that alleles characteristic of both the Atlantic and the Danubian lineages are found in the population. However, Atlantic alleles dominate throughout the broodstock which is in Hardy–Weinberg equilibrium according to the investigated markers. Results indicate that the original broodstock that was introduced to the farm following its construction in 1933 was of the Atlantic lineage. Although later fish from a local stream were collected and added to the broodstock, the number of these was limited and they were almost exclusively males. Fish from this farm that are stocked by anglers can have a significant genetic impact on trout populations of natural streams.     

PCR survey for Paramoeba perurans in fauna,environmental samples and fish associated with marine farming sites for Atlantic salmon (Salmo salar L.)

Amoebic gill disease (AGD) caused by the amoeba Paramoeba perurans is an increasing problem in Atlantic salmon aquaculture. In the present PCR survey, the focus was to identify reservoir species or environmental samples where P. perurans could be present throughout the year, regardless of the infection status in farmed Atlantic salmon. A total of 1200 samples were collected at or in the proximity to farming sites with AGD, or with history of AGD, and analysed for the presence of P. perurans. No results supported biofouling organisms, salmon lice, biofilm or sediment to maintain P. perurans. However, during clinical AGD in Atlantic salmon, the amoeba were detected in several samples, including water, biofilm, plankton, several filter feeders and wild fish. It is likely that some of these samples were positive as a result of the continuous exposure through water. Positive wild fish may contribute to the spread of P. perurans. Cleaner fish tested positive for P. perurans when salmon tested negative, indicating that they may withhold the amoeba longer than salmon. The results demonstrate the high infection pressure produced from an AGD‐afflicted Atlantic salmon population and thus the importance of early intervention to reduce infection pressure and horizontal spread of P. perurans within farms.     

Experimental infection of Atlantic halibut, Hippoglossus hippoglossus L., yolk-sac larvae with infectious pancreatic necrosis virus: detection of virus by immunohistochemistry and in situ hybridization

Three different concentrations (10⁷, 10⁵ and 10³ TCID₅₀ ml^-1) of infectious pancreatic necrosis virus (IPNV) serotype Sp isolated from Atlantic halibut, Hippoglossus hippoglossus L., were used to bath-challenge Atlantic halibut yolk-sac larvae. The larvae challenged with 10⁷ TCID₅₀ ml^-1 suffered significantly higher cumulative mortality than the other challenged groups and the control group, and affected individuals displayed necrosis of the intestine, liver and kidney. In larvae from the groups challenged with 10⁷ and 10⁵ TCID₅₀ ml^-1, IPNV was detected by immunohistochemistry and in situ RNA/DNA hybridization in the intestine, liver and kidney. In addition, some individuals stained IPNV-positive in the heart and eye/brain region. Detection by in situ hybridization did not appear to be more sensitive than immunohistochemistry. However, background staining was virtually absent in comparison with immunohistochemistry, and the staining seemed to be more distinctly localized to the cytoplasm of infected cells. The results show that farmed halibut yolk-sac larvae can be infected by IPNV immediately after hatching, with resulting high mortality. As the larvae are not immunologically mature at this stage of development, vaccination is not recommended.     

Use of encounter data to model spatio‐temporal distribution patterns of endangered smalltooth sawfish,Pristis pectinata,in the western Atlantic

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Experimental infection of Atlantic halibut,Hippoglossus hippoglossus L., yolk-sac larvae with infectious pancreatic necrosis virus: detection of virus by immunohistochemistry and in situ hybridization*

Three different concentrations (10⁷, 10⁵ and 10³ TCID₅₀ ml^?1) of infectious pancreatic necrosis virus (IPNV) serotype Sp isolated from Atlantic halibut, Hippoglossus hippoglossus L., were used to bath-challenge Atlantic halibut yolk-sac larvae. The larvae challenged with 10⁷ TCID₅₀ ml^?1 suffered significantly higher cumulative mortality than the other challenged groups and the control group, and affected individuals displayed necrosis of the intestine, liver and kidney. In larvae from the groups challenged with 10⁷ and 10⁵ TCID₅₀ ml^?1, IPNV was detected by immunohistochemistry and in situ RNA/DNA hybridization in the intestine, liver and kidney. In addition, some individuals stained IPNV-positive in the heart and eye/ brain region. Detection by in situ hybridization did not appear to be more sensitive than immunohistochemistry. However, background staining was virtually absent in comparison with immunohistochemistry, and the staining seemed to be more distinctly localized to the cytoplasm of infected cells. The results show that farmed halibut yolk-sac larvae can be infected by IPNV immediately after hatching, with resulting high mortality. As the larvae are not immunologically mature at this stage of development, vaccination is not recommended.     

An attempt at crossbreeding Atlantic salmon and pink salmon (preliminary report)

The results of artificial crossbreeding of Salmo salar and Oncorhynchus gorbuscha of the Kola river (Murmansk region) are discussed. Observations are presented on developmental characteristics of the hybrid and the Atlantic salmon parent, as well as karyological data on hybrid and control embryos.Crosses of Atlantic salmon and pink salmon of the Kola river produced non-viable embryos with various morphological anomalies. The diploid chromotype of the hybrid ranged from 52 to 56. The number of two-armed chromosomes in the karyotype of the hybrid embryos was greater than that in the karyotype of Atlantic salmon. It is concluded that crosses of Atlantic salmon and pink salmon could not occur in nature.     

Enhanced production of the sea urchin Paracentrotus lividus in integrated open-water cultivation with Atlantic salmon Salmo salar

Survivorship and performance was investigated for two size classes of the sea urchin Paracentrotus lividus grown adjacent to open-water Atlantic salmon (Salmo salar) cultivation on the north-west coast of Scotland. Juvenile P. lividus were maintained for 12 months at 0 m, 50 m and 2.5 km from the mariculture activity. The sea urchins at the 0 m station showed higher survivorship than at the 50 m and 2.5 km stations and had significantly greater test diameter than at the 50 m station. Only urchins at 0 m developed gonads and, although small, these were of acceptable or excellent colouration in terms of their marketability. Adult P. lividus were maintained for 3 months at 0 m and 15 m from the mariculture activity, either with or without additional macroalgae Laminaria spp. Sea urchins at the 0 m station and fed additional macroalgae showed higher gonadal growth than sea urchins at the 15 m station held without additional food. Gonad colouration in the adult urchins, irrespective of the treatment, was acceptable or excellent. The 0 m station also received significantly greater quantities of particulate organic matter (POM) than the other stations in both the juvenile and adult experiments. At the 0 m station, the presence of the fatty acid 22:1n-11 and elevated levels of the long chain n-3 fatty acid DHA 22:6n-3 in the gonadal tissue of the urchins confirmed the consumption of fish farm derived POM and highlighted the potential human health benefits from consuming sea urchin roe.The results show that P. lividus can thrive in the salmon culture environment and suggests that the integration of P. lividus with Atlantic salmon can provide a viable means to culture this species, even at this northerly latitude. Salmon-sea urchin co-culture would enable fish farms to diversify into producing a second highly valuable product and would reduce the increasing worldwide pressure of sea urchin fisheries on wild stocks.     

Feeding regime does not influence lysine utilisation by Atlantic salmon, Salmo salar L., parr

This study investigated whether the efficiency of lysine utilisation for liveweight gain, protein gain and lysine gain was affected by feeding regime in Atlantic salmon (Salmo salar L.) parr. Twelve diets containing from 10.15 to 20.79 g dietary digestible lysine kg^− 1 were hand-fed either to satiation or to a controlled fixed ration. The controlled ration was set to equal the feed intake of the basal (lowest) lysine diet so that any growth above that of the group fed the basal diet at the controlled ration was due entirely to the additional dietary lysine. For both feeding regimes, with increasing dietary lysine there were significant linear increases in liveweight gain (P < 0.001) and in protein (P < 0.001) and lysine (P < 0.01) concentration of liveweight gain. Increasing dietary lysine resulted in a significant (P < 0.001) linear increase in feed intake at satiation. Efficiency of lysine utilisation for liveweight gain above maintenance lysine intake was significantly (P < 0.005) higher at satiation than for the controlled ration: 47.7 compared to 34.9 mg liveweight gain per mg digestible lysine intake, respectively. However, feeding regime had no significant effect on the efficiency of lysine utilisation for protein or lysine gain. This study demonstrated that feed intake does not influence the efficiency of lysine utilisation for protein or lysine gain in Atlantic salmon parr. The change in weight-specific lysine composition suggests a robust physiological mechanism maintaining the efficiency of lysine utilisation for lysine gain in Atlantic salmon parr.