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1.
Megalocytivirus belongs to the Iridoviridae family and is known to affect finfish. Megalocytivirus epizootics have been reported to occur in several cultured fish species in China; however, no megalocytivirus associated with rock bream, Oplegnathus fasciatus (Temminck & Schlege), has been documented. In this study, we characterized for the first time a megalocytivirus, rock bream iridovirus (RBIV)‐C1, detected in cultured rock bream in a fish farm in China that had been inflicted with a high‐mortality disease outbreak. Sequence analysis of three conserved genes showed that RBIV‐C1 shares over 90% overall identities with a number of known megalocytiviruses. Electron microscopic examination revealed RBIV‐C1 as hexagonal particles similar to those reported for megalocytiviruses. In vivo infection study indicated that, following inoculation into rock bream, RBIV‐C1 induced 100% mortality and upregulated the expression of Mx, IL‐1β and IL‐8. The infected fish exhibited pathological signs similar to those observed in naturally diseased fish. Furthermore, studies in a turbot (Scophthalmus maximus L.) model indicated that RBIV‐C1 induced acute infection in turbot that led to 100% mortality. These results indicate that RBIV‐C1 is highly virulent to rock bream as well as turbot and that RBIV‐C1 is closely related to a number of previously reported megalocytivirus and likely a genetic variant of the latter.  相似文献   

2.
The genus Megalocytivirus is known to infect a wide range of cultured marine fish. In this study, we examined the pathogenicity of FLIV (Megalocytivirus from olive flounder, genotype III) and RBIV (Megalocytivirus from rock bream, genotype I) to their homologous and heterologous host species. Olive flounder (7.5 ± 1.3 cm) injected with FLIV [major capsid protein (MCP) gene copies, 6.8 × 103–6.5 × 106/fish] at 24 °C did not die until 90 days post‐infection (dpi). The average virus replication in the spleen peaked (1.27 × 106/fish) at 20 dpi. Rock bream (6.5 ± 1.5 cm) injected with FLIV (8.8 × 105 and 6.5 × 106/fish of MCP copies) showed no mortality until 50 dpi. The rock bream that survived after FLIV infection were rechallenged with RBIV at 50 dpi had 100% mortality, showing that there is no cross‐protection between FLIV and RBIV. Temperature shifting (26 °C and 20 °C at 12 h intervals) did not cause FLIV‐specific mortality into olive flounder, but higher virus copies were observed in the fish exposed to higher stocking density. This study demonstrates that FLIV and RBIV have different antigenic and pathogenic characteristics and that FLIV has low pathogenicity to olive flounder.  相似文献   

3.
As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL‐8 and COX‐2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL‐1β, IL‐8 and COX‐2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)‐induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation‐related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.  相似文献   

4.
The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.  相似文献   

5.
Scuticociliatosis is a devastating and intractable protozoal disease in olive flounder, leading to a significant loss throughout the year. This study aimed to investigate a systemically effective antiscuticociliatosis agent for olive flounder for better absorption into the infected internal organs. The in vitro and in vivo antiscuticociliatosis effects of clioquinol (CQ) were examined after screening 30 biocidal agents against the highly pathogenic scuticociliate Miamiensis avidus. CQ was the most potent in vitro drug of those tested against cultured M. avidus. CQ was the least toxic in healthy olive flounder among the drugs that exhibit high potencies. In olive flounder, a single intramuscular injection of 40 mg/kg CQ significantly reduced mortality caused by artificial infection with M. avidus, and 10–20 mg/kg CQ increased fish survival times. CQ was also effective in naturally infected scuticociliatosis. Ciliate cell numbers were lower when CQ was injected in most organs, including the brain. CQ was well absorbed by the internal organs after intramuscular injection. This study suggests that CQ can be considered as a potential antiscuticociliatosis agent for systemic administration in olive flounder.  相似文献   

6.
Rock bream iridovirus (RBIV) causes huge losses, especially in rock bream Oplegnathus fasciatus. Rock bream injected with RBIV and held at 29, 26, 23 or 20 °C had 100% mortality. Conversely, all infected fish held at 17 °C survived even after the temperature was progressively increased to 26 °C at 100 dpi. Rock bream exposed to virus and held for 2, 4 and 7 days at 23/26 °C before the temperature was reduced to 17 °C had mortality rates of 26.6/73.2%, 66.6/100% and 93.4/100%, respectively, through 100 dpi. When surviving fish had the water temperature increased from 17 to 26 °C at 100 dpi, they did not exhibit signs of disease and had low virus copy numbers (below 103). To investigate the development of a protective immune, rock bream were infected with RBIV and held at 23 °C before shifting the water temperature to 17 °C at 4 dpi. All injected fish survived until 120 dpi. While 100% of the previously unexposed fish died, 80.2% of the previously infected fish survived. When the survivors were rechallenged again at 160 dpi, no further mortality occurred. The high survival rate of fish following rechallenge with RBIV indicates that protective immunity was established in the surviving rock bream.  相似文献   

7.
This study tested the onion powder (OP) supplementation in the diet of the olive flounder on the growth, body composition, and lysozyme activity. Thirty‐five fish averaging 5.1 g were randomly stocked into 18 individual 180‐L flow‐through tanks. A commercially available OP was used as a dietary additive. Six experimental diets were prepared to contain OP at the concentrations of 0, 0.5, 1, 2, 3, and 5% for diets OP‐0, OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5, respectively. After the 8‐wk feeding trial, 20 fish from each tank were infected with Edwardsiella tarda and mortality was monitored for the following 96 h. No distinctive improvement in survival, weight gain, or feed efficiency of fish was observed at the end of the 8‐wk feeding trial. Lysozyme activity in fish fed the OP‐0.5 diet was higher than that of fish fed the OP‐0, OP‐1, OP‐2, OP‐3, and OP‐5 diets. The cumulative mortality of fish fed the OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5 diets was lower than that of fish fed the OP‐0 diet at 72 h after E. tarda infection. Dietary inclusion of 0.5% OP was effective at improving lysozyme activity of fish, and OP seemed to be an effective immunostimulant to lower mortality upon E. tarda infection.  相似文献   

8.
Pathological characteristics of olive flounder Paralichthys olivaceus experimentally infected with Streptococcus parauberis were studied. Various stressful conditions, aeration and netting stress in particular, led to induced mortality by S. parauberis. Netting stress-induced mortality was positively correlated to bacterial dose and stressful conditions. Inflammation of the heart and pericarditis was the major pathological change observed in olive flounder experimentally infected with S. parauberis. During the infected period, the number of bacteria in the infected olive flounder was recorded over time. S. parauberis remained in all fish organs tested, especially in the heart and brain.  相似文献   

9.
Turbot iridovirus (TBIV), a member of the genus Megalocytivirus in the family Iridoviridae, was isolated from diseased turbot, Psetta maximus (L.), in Korea in 2003. In this study, experimental infection of turbot, Japanese flounder, Paralichthys olivaceus (Temminck & Schlegel), and rock bream, Oplegnathus fasciatus (Temminck & Schlegel), with TBIV was performed to evaluate the viral susceptibility of these fish species. After virus exposure, the mortalities of turbot reared at 22 and 25 degrees C were 60% and 100%, respectively, suggesting that TBIV is the causative agent of the mass mortality of turbot that occurred in Korea in 2003. Moreover, TBIV was detected in Japanese flounder and rock bream by polymerase chain reaction after experimental infection (26 days post-inoculation) despite no viral pathogenicity in these fish, suggesting that these two fish species are also susceptible to the virus. It is possible that horizontal transmission of TBIV occurs among these three fish species because turbot is routinely cultured with Japanese flounder and rock bream in Korea.  相似文献   

10.
This experiment was conducted to study the effects of different dietary levels of vitamin C, L‐ascorbyl‐2‐polyphosphate (ASPP), on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Fish were fed one of six semi‐purified diets containing an equivalent of 0, 25, 50, 75, 150, or 1500 mg ascorbic acid (AA) kg?1 diet (C0, C25, C50, C75, C150 or C1500) in the form of ASPP for 12 weeks. Weight gain (WG) and protein efficiency ratio (PER) of fish fed the C0 diet were significantly lower than those of fish fed the other diets (P < 0.05), and WG and PER of fish fed the C25, C50 and C75 diets were significantly lower than those of fish fed the C1500 diet (P < 0.05). Fish fed the C0 diet exhibited vitamin C deficiency symptoms such as anorexia, scoliosis, cataract, exophthalmia and fin hemorrhage at the end of the 12‐week test. After 12 weeks of the feeding trial, AA concentrations from gill, kidney, and liver of fish fed the C0, C25, C50 and C75 diets were significantly lower than those of fish fed the C150 and C1500 diets (P < 0.05). Based on broken line analyses for WG and PER, the optimum dietary levels of vitamin C were 91 and 93 mg AA kg?1 diet respectively. These findings suggest that the dietary vitamin C requirement could be 93 mg AA kg?1 diet to support reasonable growth, and greater than 150 mg AA kg?1 diet may be required for AA saturation of major tissues for juvenile olive flounder under experimental conditions.  相似文献   

11.
This study evaluated the effects of dietary fermented tuna by‐product meal (FTBM) in juvenile olive flounder, Paralichthys olivaceus. Five diets were formulated to replace fishmeal (FM) with FTBM at 0% (FTBM0), 12.5% (FTBM12.5), 25.0% (FTBM25), 37.5% (FTBM37.5), or 50% (FTBM50). After 8 wk, weight gain, specific growth rate, and feed efficiency of fish fed FTBM0 and FTBM12.5 diets were significantly higher than fish fed the other diets (P < 0.05). Also, mean cumulative survival rates (%) of fish fed the FTBM0 and FTBM12.5 diets were significantly higher than those fed FTBM50 diet at Day 9 postchallenge with Edwardsiella tarda (P < 0.05). Protein efficiency ratio of fish fed FTBM0 and FTBM12.5 diets was significantly higher (P < 0.05) than fish fed diets FTBM37.5 and FTBM50. Broken‐line regression analysis of weight gain showed an optimal FM replacement level of 10.65% with FTBM. Therefore, the optimal dietary inclusion of FTBM in juvenile olive flounder diets could be greater than 10.65% but less than 12.5% without any adverse physiological effects on fish health.  相似文献   

12.
A 12‐week feeding trial was designed to assess the probiotic potential of indigenous Bacillus amyloliquefaciens and/or Bacillus subtilis singly or in combination with Bacillus licheniformis in an extruded feed for olive flounder (Paralichthys olivaceus) juveniles. A high fish meal (FM) diet (control) and a low‐FM diet containing an alternative protein blend (30% FM replacement, FM30) were formulated. Three other experimental diets were prepared by inclusion of B. amyloliquefaciens (BA), B. subtilis (BS), or a mixture of B. amyloliquefaciens, B. subtilis, and B. licheniformis (BASL) into FM30 diet, with a final concentration of 106 CFU/g diet. Results indicated that the FM30 diet was well tolerated by flounder, and the overall performance was not affected by dietary treatments. Lysozyme activity and total immunoglobulin level were significantly reduced in flounders when fed with the FM30 diet compared with the BASL and BA diets, respectively. The Bacillus additives neither enriched the relative abundance of the corresponding Bacillus spp. in the relevant gut microbiota of olive flounder nor modulated the presumptive gene functions of the gut microbiome. Despite the absence of growth‐promoting effect, the tested probiotics could still be economically viable for use as immunostimulants in commercial flounder diets with partial FM replacement.  相似文献   

13.
We monitored feeding behavior and survival of starved juvenile olive flounder experimentally infected with the gill monogenean Neoheterobothrium hirame. Infected flounder increased amount of the time spent in the water column by 117% when trying to capture live mysids, Neomysis sp. They also showed different feeding patterns from those of uninfected fish and made fewer attacks towards prey during one feeding attempt. Although the average numbers of mysids captured by individuals were similar between infected and uninfected fish, heavily infected fish tended to catch less prey. These results indicate that N. hirame reduces the feeding efficiency of the host for capturing live prey and possibly makes them more vulnerable to predation during feeding. We could not detect any obvious difference in survival rates between uninfected, lightly and heavily infected fish during 3 months of starvation. There was no evidence that starvation makes fish more susceptible to N. hirame. The present study provides first experimental evidence that N. hirame affects feeding behavior of juvenile olive flounder and supports the idea that this parasite indirectly reducing the host’s survival and may be responsible for the recent reduction of the flounder population in Japan.  相似文献   

14.
Enzymatic hydrolysis of soy protein has the potential to be an excellent protein source in marine fish feeds. Here we assessed the effects of different inclusion levels of hydrolyzed soy protein (HSP) on growth performance, nutrient retention, body composition, and serum hormone levels in juvenile starry flounder to define HSP utilization. Five isoenergetic and isoprotein diets were formulated with 0 (control), 10, 20, 30, and 40% HSP (SP10–40) to replace 0, 25, 50, 75, and 100% soy protein concentrate (SPC), respectively. At the end of the feeding trial, fish fed diets SP20 and SP30 had significantly increased weight gain, percent weight gain, and feed efficiency (FE) compared with fish fed the control diet (P < 0.05). Diets SP20 and SP30 supported significantly higher levels of protein retention, and diet SP20 significantly increased energy retention (P < 0.05) in the fish offered these diets. In terms of nutritional composition of whole fish, feeding diets SP20 and SP30 increased crude protein content (P < 0.05) and diets SP10, SP20, and SP30 increased ash content (P < 0.05). Fish fed diets SP20 and SP30 had higher serum insulin‐like growth factor 1 (IGF‐1) level than that of the control treatment (P < 0.05). When SPC was included at 40% of the diet, 20–30% HSP (43% soluble nitrogen) is recommended to replace 50–75% SPC for maximal growth performance and FE of starry flounder.  相似文献   

15.
Two feeding trials were carried out to determine the optimum feeding rates in juvenile olive flounder, Paralichthys olivaceus, at the optimum rearing temperature. Fish averaging 5.0 ± 0.11 g (mean ± SD) in experiment 1 and 20.2 ± 0.54 g (mean ± SD) in experiment 2 were fed a commercial diet at the feeding rates of 0%, 3.0%, 4.0%, 4.25%, 4.5% and 4.75% body weight (BW) day?1 and satiation (5.52% BW day?1) in experiment 1 and 0%, 1.0%, 2.0%, 3.0% and 3.5% BW day?1 and satiation (4.12% BW day?1) in experiment 2 at 20 ± 1 °C. Both feeding trials lasted for 2 weeks. Results from experiment 1 indicated that weight gain (WG) and specific growth rate (SGR) of fish fed to satiation were significantly higher than those of fish fed at other feeding rates while feed efficiency (FE) and protein efficiency ratio (PER) of fish fed at 4.25% BW day?1 were significantly higher than those of fish fed to satiation and fish fed at 3.0% BW day?1 (< 0.05). In experiment 2 WG, SGR and PER leveled out after the feeding rate of 3.5% BW day?1 whereas FE reached a plateau at 3.0% BW day?1. anova of FE indicated that the optimum feeding rates in 5.0 and 20 g juvenile olive flounder could be 4.25% and 3.0% BW day?1, respectively. Broken line analysis of WG suggested the optimum feeding rates of 5.17% and 3.47% BW day?1 in 5.0 and 20 g fish, respectively. Therefore, these results indicated that the optimum feeding rates could be >4.25 but <5.17% BW day?1 for 5.0 g, and it could be >3.0 but <3.47% BW day?1 for 20 g size of juvenile olive flounder at the optimum rearing temperature.  相似文献   

16.
Megalocytiviruses cause high mortality diseases that have seriously impacted aquaculture, with the most frequent outbreaks occurring in East and South‐East Asia. The international trade of juvenile fish for food and ornamental aquaculture has aided the spread of these viruses, which have spread to Europe and Australia and other regions. Australian freshwater fishes were examined for susceptibility to infection with the exotic megalocytivirus, dwarf gourami iridovirus (DGIV), which belongs to a group with the type species, Infectious spleen and kidney necrosis virus (ISKNV). Fish were held at 23 ± 1 °C and challenged by intraperitoneal (IP) injection or by cohabitation with Murray cod, Maccullochella peelii (Mitchell) infected with DGIV. A species was deemed to be susceptible to DGIV based on evidence of viral replication, as determined by qPCR, and megalocytic inclusion bodies observed histologically. Horizontal transmission occurred between infected Murray cod and golden perch, Macquaria ambigua (Richardson), Macquarie perch, Macquaria australasica (Cuvier) and Murray cod. This indicated that DGIV shed from infected fish held at 23 °C can survive in fresh water and subsequently infect these naïve fish. Further, DGIV administered IP was highly pathogenic to golden perch, Macquarie perch and Murray cod. Compared to these species, the susceptibility of southern pygmy perch, Nannoperca australis (Gunther) was lower. Freshwater catfish (dewfish), Tandanus tandanus (Mitchell), were not susceptible under the experimental conditions based on the absence of clinical disease, mortality and virus replication. This study showed the potential risks associated with naïve and DGIV‐infected fish sharing a common water source.  相似文献   

17.
This study was conducted to investigate the influence of dietary distillers dried grain (DDG) level on growth and body composition of juvenile olive flounder (Paralichthys olivaceus). Five diets (designated as DDG0, DDG7, DDG14, DDG21 and DDG28) were prepared to contain 0, 70, 140, 210 and 280 g kg?1 DDG. Three replicate groups of fish averaging 11.6 ± 0.1 g were fed one of the diets for 8 weeks. Weight gain of fish fed diets containing 70–280 g kg?1 DDG was not different to that of fish fed DDG0 diet (P > 0.05). Feed efficiency of fish fed DDG21 and DDG28 diets was lower than that of fish fed DDG0 diet (< 0.05). The antioxidant enzyme and digestive enzyme activities were not affected by dietary DDG levels. The results of this experiment suggested that DDG is a valuable ingredient as a candidate to replace wheat flour in the diet and could be used up to 280 g kg?1 for the growth of olive flounder. On the other hand, if lower feed efficiency of fish fed 210–280 g kg?1 DDG is considered, up to 140 g kg?1 DDG in diet is recommended for optimum feed utilization of fish.  相似文献   

18.
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 13758T. 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.  相似文献   

19.
This experiment was conducted to determine the optimum dietary protein level for juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel) fed a white fish meal and casein‐based diets for 8 weeks. Olive flounder with an initial body weight of 4.1 ± 0.02 g (mean ± SD) were fed one of the six isocaloric diets containing 35%, 45%, 50%, 55% and 65% crude protein (CP) at a feeding rate of 4–5% of wet body weight on a dry‐matter basis to triplicate groups of 20 fish per aquarium. After 8 weeks of feeding, per cent weight gain (WG) and feed efficiency ratios of fish fed the 55% CP diet were not significantly higher than those from fish fed the 50% and 65% CP diets, but significantly higher than those from fish fed the 35% and 45% CP diets. Fish fed the 50%, 55% and 65% CP diets had significant higher specific growth rates than did fish fed the 35% and 45% CP diets; however, there was no significant difference among fish fed the 50%, 55% and 65% CP diets. The protein efficiency ratio was inversely related to the dietary protein level; that is, maximum efficiency occurred at the lowest dietary protein level. Broken‐line model analysis indicated that the optimum dietary protein level was 51.2 ± 1.8% for maximum weight gain in juvenile olive flounder. The second‐order polynomial regression analysis showed that the maximum WG occurred at 57.7% and it revealed that the minimum range of protein requirement was between 44.2% and 46.4%. These findings suggest that the optimum dietary protein level for maximum growth could be greater than 46.4%, but less than 51.2% CP in fish meal and casein‐based diets containing 17.0 kJ g?1 energy for juvenile olive flounder.  相似文献   

20.
In 2005, massive mortality occurred in olive flounder Paralichthys olivaceus farms in Korea, and five isolates were collected from diseased fish. In this study, microbiological and pathogenic characteristics of these isolates were studied. The isolates gave negative results in lysine and ornithine decarboxylase, ortho-nitrophenyl-??-galactoside, and citrate tests, and positive results in urease, esculinase, and nitrate reduction tests. The isolates produced acid from adipate, fructose, d-glucose, and maltose, and gave positive results in alkaline phosphatase, esterase lipase, leucine arylamidase, and naphthol-AS-BI-phosphohydrolase. According to genetic analysis, 16S rRNA gene sequences showed 98?C100?% identity with both Vibrio scophthalmi and V. ichthyoenteri. The dnaJ gene sequences presented a higher identity with V. scophthalmi than with V. ichthyoenteri. Thus, the isolates were identified as V. scophthalmi. Pathogenicity of the five isolates in olive flounder was different and LD50 values were from 106 to 108?CFU/g fish. Symptoms included darkening of skin, hemorrhage of liver and intestine, ascites, and distended abdomen. Histopathological changes included hemopoiesis dilatation and epithelial hyaline droplets in kidney, macrophage infiltration and ellipsoid dilatation in spleen, vascular dilatation, submucosal edema, and serosa inflammation of intestine. Cumulative mortality was 25?% for fish singly infected by isolate A19008 or Streptococcus parauberis, and increased to 87.5?% in super-infection group with these two pathogens.  相似文献   

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