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1.
An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV) of shrimp in different organs of freshwater rice‐field crab, Paratelphusa hydrodomous (Herbst), using bioassay, PCR, RT‐PCR, ELISA, Western blot and real‐time PCR analyses, and also to use this crab instead of penaeid shrimp for the large‐scale production of WSSV. This crab was found to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT‐PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real‐time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production.  相似文献   

2.
为探究浙江省舟山市某养殖池塘中三疣梭子蟹(Portunus trituberculatus)暴发疾病的病因, 应用组织病理学、 分子生物学和荧光原位杂交等技术手段, 对患病三疣梭子蟹组织进行检验。研究发现, 患病三疣梭子蟹的临床症状主要表现为食欲下降, 行动迟缓, 鳃水肿; 光学显微镜下观察鳃及血淋巴液未发现寄生虫, 肝胰腺等组织中也未分离到致病菌; 采用 PCR 方法对病蟹进行血卵涡鞭虫(hematodinium)、白斑综合征病毒(white spot syndrome virus, WSSV)、青蟹双顺反子病毒(mud crab discistrovirus-1, MCDV-1)以及青蟹呼肠孤病毒(Scylla serrata reovirus, SSRV) 蟹类常见病原 PCR 检测, 结果均为阴性; 病蟹的肝胰腺、心脏、鳃等组织的病理切片中可观察到明显的细胞病变和嗜酸性包涵体; 超薄切片电镜观察显示: 病蟹的肝胰腺、心脏和鳃组织中均存在六边形病毒颗粒, 粒子直径 150 nm 左右, 与已报道的十足目虹彩病毒 1 (decapoda iridescent virus 1, DIV1)形态特征相似。采用特异性套式 PCR 检测方法对患病蟹组织样品进行 DIV1 病原检测, 所有样本均扩增出 457 bp 和 129 bp 大小的目的片段。进一步根据 GenBank 中 DIV1 的主要衣壳蛋白(major capsid protein, MCP)表达基因和三磷酸腺苷酶(ATPase)表达基因序列设计特异性引物, 均能从病蟹样品中扩增出预期大小的 MCP 和 ATPase 基因开放阅读框(open reading frame, ORF)区全长。将扩增获得的 MCP 和 ATPase 基因 ORF 区全长进行测序和同源序列比对分析, 进化树分析结果表明其与十足目虹彩病毒属(Decapodiridovirus)病毒的 MCP 和 ATPase 基因序列自然聚为一支, 判定导致此次三疣梭子蟹发病病原为 DIV1。根据原位杂交探针设计原则, 以 DIV1 的 MCP 和 ATPase 基因的保守区域为靶位点分别设计探针, 通过荧光原位杂交获得了病毒粒子在病蟹肝胰腺、心脏、肌肉和鳃组织的分布情况, 与电镜切片观察和套式 PCR 检测结果相符。研究结果可为海水养殖三疣梭子蟹十足目虹彩病毒 1 病诊断与防控提供参考。  相似文献   

3.
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4.
While the effects of ammonia on fish and prawn larvae are well documented, little is known of its effect on mud crab (Scylla serrata) (Forsskål, 1755) larvae. Two experiments were conducted in 5 L hemispherical plastic bowls, containing 3 L of ultra‐filtered and settled seawater and various larval stages of mud crab to (1) determine the acute median lethal concentration (LC50) of unionized ammonia and (2) to determine the chronic effects of unionized ammonia on survival and percentage moulting to zoea and megalop stages. The larval stages that exhibited the highest tolerance to ammonia over 24 h were zoea 1 (LC50 of 4.05 mg L?1 of unionized ammonia) and zoea 5 (LC50 of 6.64 mg L?1 of unionized ammonia). The megalop stage had the lowest total ammonia LC50 at both 24 and 48 h, making it the larval stage most susceptible to total ammonia. Exposure to 6.54 mg L?1 of unionized ammonia resulted in 100% death of all larvae within 24 h. The tolerance of S. serrata larvae to total ammonia did not appear to increase with ontogenetic development. The results indicate that the concentrations at which total ammonia produces an acute or chronic response in mud crab larvae are far higher than those experienced in current larval production systems (0–0.5 mg L?1 of total ammonia) used as industry standards in Australia.  相似文献   

5.
The significant decrease in wild mud crab population highlights the need to manage the resources and domesticate crabs. This paper presents the initial results of the domestication of mud crab Scylla serrata aimed at producing good-quality captive broodstock. The analysis of the genetic structure of the base population was done as a prerequisite for domestication. Adult S. serrata from the northern to southern parts of the Philippines (Cagayan, Camarines, Samar, and Surigao) were obtained for genetic diversity analysis and domestication. Analysis of molecular variance showed that differences in the genetic variability between the four populations were not significant. Moreover, no significant deviation from Hardy–Weinberg Equilibrium was observed in each sample population and even in pooled populations. Body weight was positively correlated with the carapace width. Second spawning occurred 41–46 days after the first spawning and 34 days from second to third spawning. However, there was a decrease in the number of zoea in repeat spawnings. Twenty-four first-generation (F1) families were produced from the four sites. The duration from spawning of the base population (P0) to attainment of broodstock size F1 was 10–14 months. Four second-generation (F2) families were produced after 11–12 months. Up to the F2, crabs tested negative for six viruses: white spot syndrome virus, infectious hypodermal and hematopoietic necrosis virus, gill-associated virus, yellow head virus, Taura syndrome virus, and infectious myonecrosis virus. The reproductive performance of P0 was comparable to the succeeding generations. Several families were obtained from one population in a year. However, due to the cannibalistic behavior of crabs, more space is required for the nursery and grow-out phase. The domestication of S. serrata is the first study done on any mud crab species in the Indo-west Pacific region. The initial results would serve as guide to understand and eliminate the barriers to mud crab domestication. The breeding technology developed from this study will support the production of good-quality seedstock for farming.  相似文献   

6.
The effects of varying dietary lecithin and cholesterol levels on growth, development and survival of mud crab, Scylla serrata, megalopa were evaluated using six semi‐purified, microbound diets formulated to be iso‐energetic and containing three levels of supplemental lecithin (0, 20 and 40 g kg−1 diet dry weight) and two levels of supplemental cholesterol (0 and 7 g kg−1 diet dry weight). Fifteen megalopa were reared individually in each treatment and the nutritional value of diets was assessed on basis of mean dry weight and mean carapace width of newly settled first crab stage, as well as development time to the first crab stage and overall survival. A significant interaction between supplemental dietary lecithin and supplemental dietary cholesterol was found for final mean dry weight of newly settled crabs, and highest survival (60%) was recorded for megalopa fed diets containing the highest levels of dietary lecithin (39.7–44.1 g kg−1) (diet 5 and 6) regardless of whether diets were supplemented with cholesterol; this rate of survival was identical to that of megalopa fed live Artemia nauplii. The results indicate that supplemental dietary cholesterol may not be essential for mud crab megalopa when fed diets containing sufficient levels of supplemental dietary phospholipids.  相似文献   

7.
白斑综合征病毒(WSSV)在拟穴青蟹体内增殖的研究   总被引：1，自引：0，他引：1  
拟穴青蟹(Scylla paramamosain)因肉质鲜美、生长快速而成为我国主要的海水养殖经济蟹类,但是随着养殖规模的不断扩大,病害问题也越发严重。本研究通过流行病学调查和定量PCR跟踪检测的方法,研究了拟穴青蟹对白斑综合征病毒(WSSV)的易感性和WSSV在拟穴青蟹体内的增殖情况。结果表明,拟穴青蟹是WSSV的自然宿主,自然携带率为8.47%。WSSV可通过口服途径感染拟穴青蟹,并在拟穴青蟹体内快速增殖,注射感染5 d后,病毒量达到感染1d时的1.1×109倍,当病毒累积到一定量后即可致死拟穴青蟹。研究表明,拟穴青蟹是WSSV的天然宿主,在虾蟹混养过程中可以通过摄食WSSV感染虾而带毒或发病,并因此成为WSSV传播媒介,从而对虾蟹混养条件下WSSV的防治效果产生重要影响。  相似文献   

8.
White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to ammonia, using a TaqMan‐based real‐time PCR, and their subsequent susceptibility to WSSV. Shrimp were exposed to different levels of total ammonia nitrogen (TAN) (8.1, 3.8 and 1.1 mg L?1) for 10 days and challenged with WSSV by feeding WSSV‐positive shrimp. WSSV was detected simultaneously in haemolymph, gills and pereopods at four hours post‐infection. The TaqMan real‐time PCR assay showed a highly dynamic detection limit that spanned over 6 log10 concentrations of DNA and high reproducibility (standard deviation 0.33–1.42) and small correlation of variability (CV) (1.89–3.85%). Shrimp exposed to ammonia had significantly higher (P < 0.01) WSSV load compared to the positive control, which was not exposed to ammonia. Shrimp exposed to 8.1 mg L?1 of TAN had the highest (P < 0.01) WSSV load in all three organs in comparison with those exposed to 3.8 and 1.1 mg L?1 of TAN. However, haemolymph had significantly higher (P < 0.01) viral load compared to the gills and pereopods. Results showed that shrimp exposed to ammonia levels as low as 1.1 mg L?1 (TAN) had increased susceptibility to WSSV.  相似文献   

9.
Predicting growth is critical in aquaculture, but models of growth are largely missing for mud crab species. Here, we present the first model of natural growth in juvenile and adult mud crabs Scylla serrata from East Africa using a stepwise growth function based on data on intermoult periods and growth at moult from field mark‐recapture, pond and laboratory studies. The results showed a sigmoid growth pattern in carapace width and suggest that S. serrata in East Africa will reach 300 g and sexual maturity ~9.9 months after settlement, and a commercial size of 500 g after 12.4 months. Analyses of the literature identified several issues with the common praxis to compare standard growth measures between aquaculture studies with different initial size or growing periods. Using the new growth function to estimate the proportional difference between modelled and obtained growth as an alternative method, we show that growth rates of S. serrata cultured in cage systems, which are dominant in East Africa, was <40% of the estimated natural growth and growth obtained in pond systems. The analysis also indicated that growth rates of S. serrata in Southeast Asia was over 50% higher compared with similar culture systems in East Africa, and that different species of mud crabs had large differences in growth rates. This study shows that growth in the present mud crab aquaculture systems in East Africa is below their expected potential. Further work is needed to identify the factors behind this observation.  相似文献   

10.
An attempt was made to determine the replication efficiency of hepatopancreatic parvo‐like virus (HPV) of shrimp in different organs of freshwater rice‐field crab Paratelphusa hydrodomous (Herbst) using bioassay, PCR, RT‐PCR, ELISA, Western blot and q‐PCR analyses. Another attempt was made to use this crab as an alternative to penaeid shrimp for the large‐scale production of HPV. This crab was found to be highly susceptible to HPV by intramuscular injection. The systemic HPV infection was confirmed by PCR and Western blot analyses in freshwater crab. The expression of capsid protein gene in different organs of infected crab was revealed by RT‐PCR analysis. Indirect ELISA was used to quantify the capsid protein in different organs of the crab. The copy number of HPV in different organs of the infected crab was quantified by q‐PCR. The results revealed a steady decrease in CT values in different organs of the infected crab during the course of infection. The viral inoculum that was prepared from different organs of the infected crab caused significant mortality in post‐larvae of tiger prawn, Penaeus monodon (Fabricius). The results revealed that this rice‐field crab could be used as an alternative host for HPV replication and also for large‐scale production of HPV.  相似文献   

11.
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12.
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13.
Brackishwater pond culture has been a major factor in mangrove loss in Southeast Asia, hence, the need to develop environment‐friendly technologies such as mud crab Scylla (Portunidae) culture in mangrove pens exists. This study evaluated the effects of mud crab netpen systems in central Philippines on mangrove macroflora, and the replacement of dietary fish with low‐cost pellets. Wild or hatchery‐sourced Scylla olivacea and Scylla serrata were stocked at 0.5–0.8 m−2 in 167–200 m2 nylon netpens (2.3 cm stretched mesh) in Avicennia‐dominated mangrove habitats. The feeding treatments were: (A) Zarraga: (1) no feeding (natural productivity), (2) no feeding for 1 month+supplementary feeding, (3) fish biomass and (4) low‐cost pellets, and (B) Batan: (1) fish biomass and (2) pellets+fish biomass. Feeds were given ad libitum twice daily. Growth and survival rates of S. olivacea in Zarraga pens were not significantly different among treatments, although crabs fed fish biomass had the highest survival, body weight and production. Similarly, growth and survival of S. serrata were not significantly different between the Batan treatments. Economic analysis of the latter gave a 38.5% return on investment (ROI) and 2.6 years payback period (PP) for pellets+fish biomass treatment compared with 27.5% ROI and 3.6 years PP for fish alone. Sensitivity analysis showed an improved economic performance of the pellets+fish biomass treatment by increasing the survival rate. Evaluation of mangrove community structure showed that crab culture reduced species diversity, numbers and biomass of seedlings and saplings, but not of mangrove trees. Therefore, mud crab pen culture is recommended for mangrove sites with mature trees, but not seedlings and saplings, and low‐cost pellets can reduce dependence on fish biomass.  相似文献   

14.
对虾WSSV病是亚洲对虾养殖业中的一个棘手问题。本研究采用Kimura引物 ,用PCR技术对不同生长期的中国对虾 (Penaeuschinensis)进行了WSSV的检测 ,同时也检测了对虾发病时养殖池中多见的野生厚蟹 (Helicesp .)和矛尾刺虎鱼 (Acanthogobiushasta)。检测结果表明 :分别在检测的 5尾亲虾中的 1尾 ,6尾仔虾中的 1尾 ,5尾稚虾中的 3尾及所检测的 5尾病虾和 2只厚蟹中获得到 982bp的PCR扩增产物 ,说明为WSSV感染阳性。在检测的 2尾矛尾刺虎鱼中均未获得PCR扩增产物 ,说明为WSSV感染阴性。在亲虾、虾苗以及虾池内的野生厚蟹中检测到WSSV感染的阳性结果表明 :WSSV感染的亲虾有可能是病毒的储主 ,WSSV感染的野生厚蟹有可能是病毒中间宿主或病毒的携带者 ,它们在对虾WSSV病的感染、传播中起了重要的作用  相似文献   

15.
Experimental studies were conducted by injecting or feeding white spot syndrome virus (WSSV) derived from infected shrimp, Penaeus monodon (Fabricius), collected from the south-east coast of India, to five species of shrimp, two species of freshwater prawns, four species of crabs and three species of lobsters. All species examined were susceptible to the virus. Experimental infections in the shrimp had the same clinical symptoms and histopathological characteristics as in naturally infected P. monodon . A cumulative mortality of 100% was observed within 5–7 days in shrimp injected with WSSV and 7–9 days in shrimp fed with infected tissue. Two species of mud crab, Scylla sp., survived the infection for 30 days without any clinical symptoms. All three species of lobsters, Panulirus sp., and the freshwater prawn, Macrobrachium rosenbergii (De Man), survived the infection for 70 days without clinical symptoms. However, bioassay and histology using healthy P. monodon revealed that crabs, prawns and lobsters may act as asymptomatic carriers/reservoir hosts of WSSV. This is the first report to suggest the carrier/reservoir capacity of these hosts through histological and bioassay evidences. Ultrastructural details of the virus in experimentally infected shrimp, P. vannamei , (Boone), were also studied.  相似文献   

16.
This is an initial paper in a series of overviews of biological research and aquaculture development of the mud crab, Scylla serrata, in China. Results of experimental ecological studies on mud crabs are reported here. As a result of these experimental studies, results that are important for mud crab culture were also discovered and these include, methods to condition and manage broodstock, determination of ecological conditions that are suitable for embryonic development, and the influence of temperature, salinity, diet and larval density on development and survival of larvae. Results of this work will be useful in establishing a good method for artificial mass culture of larvae.  相似文献   

17.
The effects of ammonia and nitrite on survival, growth and moulting were investigated in juvenile tiger crab, Orithyia sinica (carapace length 3.91±0.15 mm, carapace width 3.84±0.23 mm, n=440), after 30 days exposure to ammonia‐N (0, 20, 50, 100 and 150 mg L?1) and nitrite‐N (0, 50, 100, 150, 200 and 250 mg L?1) using a continuous flow system. Survival rates of tiger crab exposed to ammonia and nitrite decreased linearly with the exposure time and concentration. The growth rate of tiger crab exposed to 50, 100 and 150 mg L?1 ammonia was significantly lower than that of control crabs. The growth rate of tiger crab exposed to nitrite decreased at 150, 200 and 250 mg L?1 nitrite. During the ammonia and nitrite exposure, the intermoult period of the juveniles of tiger crab O. sinica was shortened between the first and second moult, and the number of moulting of crabs exposed to a higher concentration were significantly higher than that of control crabs.  相似文献   

18.
It is yet unclear whether sub‐lethal ammonia‐N levels cause irreparable damage to aquatic crustaceans, or if recovery is possible, the potential factors involved. The aim was to investigate the effect of 0.706 and 2.798 mmol L?1 ammonia‐N exposure on the haemolymph osmolality, Na+, K+, Ca2+, pH, ammonia‐N, total haemocyte counts (THC) and gill histopathology of Portunus pelagicus juveniles at 0, 3, 6, 12, 24 and 48 h respectively. Following 48 h, crabs were transferred to pristine seawater allowing a recovery period up to 96 h and similarly measured. In addition moribund crabs, induced from lethal ammonia‐N levels of 7.036 and 10.518 mmol L?1, were measured for haemolymph osmolality/ions and pH levels. The results demonstrate that despite severe gill damage within 6‐ and 1 h of 0.706 and 2.798 mmol L?1 ammonia‐N exposure, respectively, no significant change (P>0.05) in the haemolymph osmolality, Na+, K+, Ca2+ or pH levels occurred or by ammonia‐N‐induced morbidity. Although the gills can completely recover within 24 and 48 h post exposure to 0.706 and 2.798 mmol L?1 ammonia‐N, respectively, likely facilitated by significant haemocyte increases (P<0.05) within the haemolymph and gill lamellae, dependent factors were the previous ammonia‐N concentration and recovery duration while individual variability was also noticed.  相似文献   

19.
The fishery biology of mud crabs Scylla spp. was examined using baited traps and gill nets from September 2001 to August 2005 at Iriomote Island, Japan. To elucidate the growth of the crabs, artificially produced S. serrata juveniles were released and recaptured at the study site. The sizes at which 50% of females and males of S. serrata reached sexual maturity (SM50) were estimated as an external carapace width (ECW) based on the morphology of the abdomen and the chela respectively. Two species, S. serrata and S. olivacea, were identified in the area with S. serrata being the dominant species (>95% of the catch). Changes in the mean ECW and the results of the release and recapture experiments suggested that the recruitment of young crabs to the fishery occurred from December/January to April/May. The SM50 of females and males occurred at 132.4 and 150.7 mm ECW respectively. The body size composition of S. serrata revealed that immature crabs comprised approximately 40 and 65% of the catch for females and males respectively. To maintain a sustainable fishery for S. serrata, a minimum landing size based on the SM50 estimates should be implemented as a fishing regulation.  相似文献   

20.
The VP 28 gene encoding a structural envelope protein of the white spot syndrome virus (WSSV) was cloned into a pET32a(+) expression vector for the production of the recombinant VP28 protein. A purified recombinant protein of 39.9 kDa size was used for polyclonal antibody production in rabbit. Specific immunoreactivity of the rabbit anti rVP28 antiserum to the viral antigen was confirmed by a Western blot. The specificity of this polyclonal anti‐rVP28 antiserum to detect the presence of the virus in WSSV‐infected Penaeus monodon was verified using a immunodot blot assay. Immunodot blot showed a positive reaction in infected shrimp tissues with prominent colour development using 3,3′,5,5′‐tetramethylbenzidine (TMB) as a chromogenic substrate when compared with 3–3′ diaminobenzidine tetrahydrochloride (DAB). Highest signal intensities of the immunodots were observed in infected shrimp pleopod extracts and haemolymph. On comparison with polymerase chain reaction (PCR), immunodot blot could detect 76% of PCR‐positive WSSV‐infected shrimp samples. Immunodot blot was found to be equivalent to first‐step PCR sensitivity to detect WSSV particles estimated to contain 1.0 × 105 viral DNA copies.  相似文献   

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