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1.
采用电子显微镜观察和细胞培养等技术, 从湖北黄陂某养殖场患病大口黑鲈(Micropterus salmoides)体内发现并分离到一株蛙病毒。患病大口黑鲈的临床症状主要表现为体表出血、溃疡, 肝脏发白。将病鱼内脏组织匀浆超微滤液接种鳜脑细胞系(mandarin fish brain, MFB)细胞能产生典型细胞病变效应(cytopathic effect, CPE), 病毒滴度达到 108.36±0.15 TCID50/mL。细胞培养病毒的超薄切片电镜观察结果显示, 细胞质中存在大量直径约为 150 nm 左右的正六边形病毒粒子, 呈晶格排列。细胞培养病毒的人工感染大口黑鲈试验结果显示, 7 d 内试验鱼死亡率高达 100%, 其临床症状与自然发病鱼相似。采用大口黑鲈病毒(largemouth bass virus, LMBV)的特异性 PCR 检测方法对患病鲈组织样品和细胞培养病毒样品进行检测, 均能扩增出 241 bp 的单一目的条带。进一步根据 GenBank 中 LMBV 主衣壳蛋白(major capsid protein, MCP)基因序列设计特异性引物, 均能从上述样品中扩增出 1392 bp 的 MCP 基因开放阅读框(open reading frame, ORF)全长。将 MCP 氨基酸全序列进行比对, 结果显示其与 Santee-Cooper 蛙病毒、孔雀鱼病毒 6 型及大口黑鲈溃疡综合征病毒的 MCP 氨基酸序列同源性高达 100%。系统进化结果显示, 与感染鱼类的虹彩病毒科蛙病毒属病毒, 如鳜鱼蛙病毒、Santee-Cooper 蛙病毒、孔雀鱼病毒 6 型和大口黑鲈溃疡综合征病毒等聚成一支。这些结果证明, 该分离株为虹彩病毒科蛙病毒属的成员, 暂命名为大口黑鲈蛙病毒(largemouth bass ranavirus, LMBRaV)湖北株 LMBRaV-HB001。病毒敏感细胞系筛选试验结果表明, 病毒 LMBRaV-HB001 感染鲤上皮瘤细胞(epithelioma papulosum cyprinid, EPC)、草鱼性腺细胞(grass carp ovary, GCO)、大鲵肌肉细胞(giant salamander muscle, GSM)和鲫脑组织细胞(gibel carp brain, GiCB)均能产生典型 CPE, 病毒滴度可达 108.0 TCID50/mL 以上。本研究首次在湖北省养殖大口黑鲈体内分离与鉴定了 LMBRaV 病毒, 建立了病毒的细胞培养方法, 为进一步研究该病毒的传播、诊断和防控技术提供了重要参考。  相似文献   

2.
In 2013, an outbreak of ulcerative disease associated with ranavirus infection occurred in barcoo grunter, Scortum barcoo (McCulloch & Waite), farms in Thailand. Affected fish exhibited extensive haemorrhage and ulceration on skin and muscle. Microscopically, the widespread haemorrhagic ulceration and necrosis were noted in gill, spleen and kidney with the presence of intracytoplasmic eosinophilic inclusion bodies. When healthy barcoo grunter were experimentally challenged via intraperitoneal and oral modes with homogenized tissue of naturally infected fish, gross and microscopic lesions occurred with a cumulative mortality of 70–90%. Both naturally and experimentally infected fish yielded positive results to the ranavirus‐specific PCR. The full‐length nucleotide sequences of major capsid protein gene of ranaviral isolates were similar to largemouth bass virus (LMBV) and identical to largemouth bass ulcerative syndrome virus (LBUSV), previously reported in farmed largemouth bass (Micropterus salmoides L.), which also produced lethal ulcerative skin lesions. To the best of our knowledge, this is the first report of a LMBV‐like infection associated with skin lesions in barcoo grunter, adding to the known examples of ranavirus infection associated with skin ulceration in fish.  相似文献   

3.
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.  相似文献   

4.
This study investigated the use of alkaline hydrolysis at ambient temperature for inactivation of selected fish pathogens in fish tissues under conditions approximating those that are likely to be found in the aquaculture industry. Infectious salmon anaemia virus (ISAV) and Lactococcus garvieae have been determined in a previous study to be the most resistant virus and bacteria to pH 12 from a wide range of viruses and bacteria tested. They were spiked at high titres into fish extracts that were then treated with 1 m sodium hydroxide (NaOH). Viable L. garvieae was not detected in the treated fish extract after 1 h, and ISAV was not detected after 24‐h exposure. Field mortalities of Atlantic salmon, Salmo salar L., caused by infectious pancreatic necrosis virus were treated by alkaline hydrolysis at ambient temperature. The macerated fish mortalities contained a high titre of virus (3.38 × 108 TCID50 g?1) that was reduced to approximately 2.2 × 103 TCID50 g?1 after 24‐h exposure to NaOH, and virus was not detected after exposure for 48 h. The results suggest that alkaline hydrolysis at ambient temperature has potential as a biosecure treatment method for fish by‐products containing fish pathogens.  相似文献   

5.
The main objectives of this study were to approximate the maximum yield and evaluate size dispersion of stocker size largemouth bass, Micropterus salmoides reared in a semi‐closed recirculating system for 60 days. Fingerlings with an average body weight of 36.7 g were utilized for the study. An experimental system consisting of 18 square plastic tanks (165 L) equipped with a radial flow settler, a sump, a moving bed filter, a centrifugal pump, a rapid sand filter, a down‐flow oxygen saturator and a UV sterilizer was utilized for the trial. The system was operated semi‐closed, accounting for a daily exchange rate of 30–50% of total system water volume. Experimental stocking densities were 4.5, 9.1, 18.8, 36.5, 54.6 and 73 kg m?3 with three replicates per treatment. At the end of the experimental trial, largemouth bass showed acceptable feed conversion (1.00–1.48), specific growth rate (1.16–1.45% day?1) and survival rate (81.8–96.6%) in all treatments, displaying the highest performance at an initial stocking density range of 18–36 kg m?3. Based on a piecewise regression model with breakpoint analysis, maximum yield of largemouth bass fingerlings should not exceed 70 kg m?3. As stocking density increased, relatively more underweight fingerlings were produced with a higher uniformity of fatness.  相似文献   

6.
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.  相似文献   

7.
The influence of environmental variables on largemouth bass Micropterus salmoides (Lacépède) populations in 22 shallow (mean depth 1.0–2.9 m) Nebraska lakes was evaluated. Largemouth bass exhibited density‐dependent size structure and growth, but not condition. Total annual mortality averaged only 30%. Deeper lakes contained low density largemouth bass populations with a high proportion of fish >380 mm, whereas larger lakes with little submerged vegetation had faster growth. The proportion of largemouth bass >380 mm and relative abundance tended to increase with emergent vegetation coverage. More stable recruitment was evident in shallower lakes with increased emergent vegetation coverage. Strong year classes were associated with cooler September air temperatures. Largemouth bass populations exhibited density‐dependent effects in lakes up to 332 ha. Lake depth, emergent vegetation and autumn air temperatures may influence largemouth bass populations more than previously suggested.  相似文献   

8.
Activity patterns of largemouth bass in a subtropical US reservoir   总被引：1，自引：0，他引：1  
Abstract  Nine largemouth bass, Micropterus salmoides Lacepède, >1.5 kg were implanted with radio tags and tracked for 1 year in an embayment of Lake Seminole, Georgia, USA. Most largemouth bass movement was <50 m h−1 in all seasons and at all times of day. During the day largemouth bass were offshore in deeper water near large woody structures and moved little. Movement was lower during dusk and night periods, and a general movement towards shoreline areas was evident. Water depths where largemouth bass were located had bimodal distributions in all seasons and diel periods, with one mode typically at depths <1 m and another around 3 m. Home ranges were typically largest in the summer and smallest in the winter. Largemouth bass appeared to divide their time between an offshore resting area, primarily occupied during the day, and a near-shore area, where foraging presumably occurred, primarily used during low-light periods. The largemouth bass population in Lake Seminole consists of multiple overlapping subpopulations, offering the opportunity for embayment-specific management.  相似文献   

9.
Abstract –  A dietary analysis of largemouth bass ( Micropterus salmoides ), an exotic, piscivorous species, was conducted in large South Korean river systems (>third order streams, 31 sites). Micropterus salmoides larger than 100 mm exhibited intense piscivory on native Korean juvenile fishes, with levels of piscivory amongst the highest recorded globally, for native and introduced populations. Largemouth bass exhibited an ontogenetic shift in diet, showing a progressive increase in piscivory with size, typical of this species within its native range but unlike several studies on introduced largemouth bass in Europe and Africa. Sampling of fish communities at the same sites used for diet studies showed that native piscivores and the main food fish species of largemouth bass were significantly less abundant ( P  = 0.049 and 0.045, respectively) at sites where bass were present than at sites where no bass were recorded. Largemouth bass may pose a threat to the structure of fish assemblages in Korean river systems and further study of their population interactions and how to minimise their spread is needed.  相似文献   

10.
Australian bass Macquaria novemaculeata were challenged by immersion with nervous necrosis virus (NNV) at different ages and under controlled conditions to investigate factors affecting disease expression. Fish challenged at 3 weeks of age with 103 TCID50/ml and higher doses developed clinical disease; a lower dose of 102 TCID50/ml resulted in incidence below 100% and 101 TCID50/ml was insufficient to cause infection. Additionally, fish were challenged at 5, 6 and 13 weeks of age at 17 and 21°C to assess the role of the age of the host and water temperature on disease expression. Although Australian bass challenged at all ages had evidence of replication of NNV, only those challenged at 3 weeks of age (20 and 24 days post?hatch [dph]) developed clinical disease. Higher water temperature had an additive effect on disease expression in larvae challenged at 24 dph, but it did not affect the disease outcome in older fish. Finally, isolates of NNV derived from fish with clinical or subclinical disease presentations caused similar cumulative mortality and clinical signs when larvae at 24 dph were challenged, suggesting that agent variation was not responsible for variation in clinical presentation in these field outbreaks of NNV infection.  相似文献   

11.
Yellowfin sea bream (Acanthopagrus latus) is an important economic fish, which is seriously threatened by various fish viruses. In this study, a cell line designated as ALL derived from the liver of yellowfin sea bream was developed and characterized. The cell line grew well in Dulbecco's modified Eagle's medium containing 10%–20% foetal bovine serum at 28°C. Amplification of the cytochrome B gene indicated that ALL cells originated from yellowfin sea bream. The modal chromosome number of ALL cells was 48. ALL cells were efficiently transfected with pEGFP-N3 plasmids, indicating the potential application of ALL cells in exogenous gene manipulation studies. ALL cells were susceptive to three main fish viruses, including viral haemorrhagic septicaemia virus (VHSV), red-spotted grouper nervous necrosis virus (RGNNV) and largemouth bass virus (LMBV). The replication of VHSV, RGNNV and LMBV in ALL cells was confirmed by quantitative real-time polymerase chain reaction, virus titre and transmission electron microscopy assays. Moreover, ALL cells could respond to VHSV, RGNNV and LMBV infections, as indicated by the differential expression of antiviral genes involving in the innate immune response. In conclusion, the newly established ALL cell line will be an excellent in vitro platform for the study of the virus–yellowfin sea bream interaction.  相似文献   

12.
Sanguinarine (SA), with antimicrobial and antiparasitic activities against fish pathogens, exhibits great potential commercial use in aquaculture. However, little information on pharmacokinetics of SA restricts further application in aquaculture. In this study, pharmacokinetics of SA in common carp (Cyprinus carpio) following a single intraperitoneal administration [10 mg kg?1 BW (body weight)] was evaluated by high‐performance liquid chromatography (HPLC). The peak concentration (Cmax) of SA in kidney was 11.8 μg g?1, which was higher than in other tissues and plasma. The terminal half‐life in fish tissue and plasma was as follows: 42.3 h (kidney) > 37.2 h (liver) > 20.1 h (gill) > 18.8 h (muscle) > 10.9 h (spleen) > 10.0 h (plasma). Additionally, we determined the bacterial loads in tissues of common carp infected with Aeromonas hydrophila after i.p. administration of SA at 0, 5, 10 and 20 mg kg?1 BW. The results showed that i.p. administration of SA at 10 mg kg?1 BW significantly enhanced antibacterial efficacy against A. hydrophila, where the antibacterial ratio in the gill, kidney, spleen and liver on day 5 was 95.13%, 93.33%, 90.09% and 92.82%, respectively. Overall, these results suggested the potential of SA to treat A. hydrophila infection in common carp farming industry.  相似文献   

13.
雷存科  陈中元  张奇亚 《水产学报》2016,40(10):1643-1647
利用3个不同物种的水生动物细胞系,包括爪蟾肾细胞系(A6)、大鲵胸腺细胞系(GSTC)和鲤上皮瘤细胞系(EPC),分别用沼泽绿牛蛙蛙病毒(RGV)和大鲵蛙病毒(ADRV)感染,进一步研究细胞病变显微形态、病毒滴度、细胞病变与不同感染时间的相关性等。结果显示,在光镜下可见感染病毒的细胞发生病变,A6和EPC细胞肿胀或破裂;GSTC细胞收缩或聚在一起形成多层。同种水产动物细胞系对不同蛙病毒的敏感性不同,在A6、EPC和GSTC细胞中,RGV的滴度分别为10~(3.6)、10~(5.9)和10~(6.6) TCID_(50)/m L;ADRV的滴度分别为10~(4.3)、10~(5.4)和10~(6.1) TCID_(50)/m L,表明GSTC细胞系对两种蛙病毒都更敏感。研究为后续蛙病毒致病机理提供了有用的信息和重要实验材料。  相似文献   

14.
Infectious dose and shedding rates are important parameters to estimate in order to understand the transmission of infectious pancreatic necrosis virus (IPNV). Bath challenge of Atlantic salmon post‐smolts was selected as the route of experimental infection as this mimics a major natural route of exposure to IPNV infection. Doses ranging from 102 to 10?4 50% end‐point tissue culture infectious dose (TCID50) mL?1 sea water were used to estimate the minimum infectious dose for a Scottish isolate of IPNV. The minimum dose required to induce infection in Atlantic salmon post‐smolts was <10?1 TCID50 mL?1 by bath immersion (4 h at 10 °C). The peak shedding rate for IPNV following intraperitoneal challenge using post‐smolts was estimated to be 6.8 × 103 TCID50 h?1 kg?1 and occurred 11 days post‐challenge. This information may be incorporated into mathematical models to increase the understanding of the dispersal of IPNV from marine salmon sites.  相似文献   

15.
The behavioural and metabolic responses of the largemouth bass Micropterus salmoides (Lacépède) to temperature were determined to define optimal thermal conditions. The final preferendum of largemouth bass juveniles determined with acute and gravitation methods was independent of the method (29.0–28.1 °C). The displacement velocity in the horizontal thermal gradient of bass juveniles was 22.4 cm h−1 in the light phase and 22.6 cm h−1 in the dark phase. Oxygen consumption rates in the largemouth bass increased significantly (P<0.05) from 48.8 to 69.4 mg O2 Kg−1 h−1 with an increase in the acclimation temperature from 20 to 33 °C. The temperature quotient (Q10) in the juveniles was 1.37–2.00 in the range of acclimation temperatures of 26–29 and 29–32 °C. The optimum temperature range for growth calculated using Jobling's equation was 28.1–28.6 °C and for Q10 values 26–29 °C. The results are discussed in relation to the use of this information in aquaculture.  相似文献   

16.
Francisella orientalis is a highly virulent, emerging bacterium that causes mass mortalities in tilapia. This pathogen also affects numerous other warm-water fish species, including three-line grunt, hybrid striped bass and various ornamental fish. This study sheds light on two new species of fish that are susceptible to F. orientalis. Asian seabass and largemouth bass showed variable levels of susceptibility in a bacterial challenge experiment. After intraperitoneally injected with a dose of 106 CFU/fish, a total of 64.28% and 21.42% mortalities were obtained in Asian seabass and largemouth bass, respectively. Meanwhile, Nile tilapia showed acute mortality of 100%. All fish showed typical lesions of francisellosis, including multifocal granulomas in the spleen and head kidney. Immunohistochemical analysis revealed strong positive signals inside the granulomas of all fish. The bacterial recovery in solid media from infected fish was highest in Nile tilapia (85.71%), followed by Asian seabass (35.71%) and largemouth bass (21.42%). PCR results tested 100% positive for Nile tilapia, and 78.57% and 21.42% for Asian seabass and largemouth bass, respectively. In conclusion, Asian seabass and largemouth bass are susceptible to this pathogen, which warrants new management strategies when employing predation polyculture systems of these species with tilapia.  相似文献   

17.
Abstract Factors that can influence changes in lipid class concentrations in specific tissues have not been reported for largemouth bass Micropterus salmoides. We investigated patterns of lipid composition by measuring seasonal levels of total lipid and major lipid classes in the muscle and liver tissues of juvenile and adult largemouth bass and gonad tissue of adults. Fish were collected in December, February, April and July to correspond with key stages of the sexual cycle. Total lipid concentrations were substantially greater in hepatic (32.8 [female] to 56.6 [male] mg. g?1) and gonadal tissues (34.4 [F] to 204.0 [M] mg. g?1) than in muscle (7. 5 [F] to 11.4 [M] mg. g?1). Muscle lipid concentrations in juveniles were significantly higher (10.9 [F] and 11.4 [M] mg. g?1) in summer than in winter in adult muscle tissue (8.6 [F] and 9.4 [MI mg. g?1). A 44% increase in ovary lipid was observed from preparatory to prespawn stages, followed by a 78% decline after the spawn. Additionally, seasonal hepatic lipid concentrations were substantially higher in juveniles. Concentrations of lipid classes (free cholesterol [C], triacylglycerol [TAG], free-fatty acids [FFA], esterified cholesterol [CE] and phospholipid [PL]) of all tissue types varied significantly among season, sex, and maturity categories. Changes with all lipid classes in the ovary coincided with those occurring in total ovarian lipid. Mobilization and transport of TAG and PL from the liver to the ovary appeared to occur during gametogenesis. All class concentrations in juveniles were invariably higher in liver than in muscle, but seasonally variable among hepatic and gonad tissues of adults. Juvenile males showed slightly greater hepatic C concentrations than other groups. Muscle and hepatic C were generally higher in juveniles.  相似文献   

18.
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19.
Atlantic salmon smolts, previously unexposed to infectious pancreatic necrosis virus (IPNV), were placed into tanks of sea water at 10 °C. After 4 weeks, 40 fish were injected intraperitoneally (i.p.) with homogenized and filter‐sterilized kidney material obtained from salmon with clinical IPN in a marine farm in Shetland. The injected fish were cohabited with 40 untreated fish. Mortalities began in the injected fish on day 7 and reached a peak of 48% on day 14. In the cohabitation group, mortalities began on day 14 and reached a peak of 70% on day 27. The IPNV in the Shetland kidney homogenate was cultured in Chinook salmon embryo (CHSE) cells and passed twice. This cultured virus was injected i.p. into fish at various doses ranging from 10 to 107 TCID50 fish?1 4 weeks after seawater transfer. Challenge tanks contained 30 injected fish and 30 cohabitees. Mortality rates and levels were dose‐dependent. The highest dose used resulted in a similar mortality pattern as obtained with a similar dose of the Shetland kidney homogenate, indicating that virulence was retained after two passes in tissue culture. Even with the lowest dose, mortality reached 12% in the injected group and 23% in the cohabitees. The IPNV titres were high (106?109 i.u. g?1 kidney) in fish which died during the experiment and low (<105 i.u. g?1 kidney) or undetectable in surviving fish. The cultured virus (pass 3) was used in a challenge model where the population density of fish in the tanks was high (50 injected and 50 cohabitees) or low (15 injected and 15 cohabitees). In the high stocking density tank, mortalities peaked at about 35% in the injected group and at 52% in the cohabitees. In the low stocking density tank, mortalities peaked at about 40% in the injected fish but no mortality occurred in the cohabitees. However, IPNV was detected (up to 104 i.u. g?1 kidney) in 82% of cohabitees sampled on day 30. These data suggest that lethal lateral transmission of the virus is dependent on the infectious pressure from the injected group. A further trial was conducted to investigate the effect of time post‐seawater transfer on the susceptibility of post‐smolts to IPN. Groups of fish were challenged every 2 weeks from week 0–10. Few mortalities occurred at week 0 and virus titres were high in these fish. Most survivors became carriers, some with titres >106 i.u. IPNV g?1 kidney. From 2 to 10 weeks after seawater transfer, mortalities in both injected and cohabitees were substantial with viral titres >107 i.u. g?1 kidney. Survivors had lower titres and in many virus was undetectable. Throughout the experiments, moribund fish were sampled for histology and all showed typical IPN histopathology.  相似文献   

20.
Marbled eels, Anguilla marmorata (Quoy & Gaimard), cultured in Taiwan exhibited haemorrhage and mortality in January 2012. The severely diseased eels bled from the gills and showed congestion of the central venous sinus of the gill filaments and haemorrhage throughout the body similar to viral endothelial cell necrosis of eel. In this study, a novel polyomavirus (AmPyV) was isolated from the diseased eels using the AMPF cell line established from the pectoral fin of healthy marbled eels. AmPyV was found to encode a long T‐antigen orthologous gene. Phylogenetic analysis showed that AmPyV was closely related to Japanese eel endothelial cell‐infecting virus. PCR assays revealed AmPyV infection throughout the systemic organs. AmPyV proliferated in the AMPF, EK‐1 and EO‐2 cells at temperatures 25–30 °C, and the progeny virus yields were 107.0, 107.4 and 107.7 TCID50 mL?1, respectively. The purified virions were icosahedral particles, 70–80 nm in diameter. No clinical signs or mortality was observed among the eels injected with the virus; however, the virus was reisolated from the brain, eyes, kidneys, fins and gills of infected eels 2 month after injection. Our results suggest that AmPyV exhibits a latent infection. Pathogen of the disease needs to study further.  相似文献   

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