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1.
White spot disease (WSD) is an important viral disease of penaeid shrimp caused by white spot syndrome virus (WSSV). WSSV isolated from WSD outbreaks in commercial shrimp (Penaeus monodon) farms in India were propagated in the laboratory in healthy shrimp. The virus was purified from the infected tissues by sucrose gradient centrifugation. The VP28 was electroeluted from SDS-PAGE gels and was used to immunize Balb/c mice to produce hybridomas secreting monoclonal antibodies (MAb) against WSSV. A total of five hybridoma clones secreting MAbs to VP28 were produced. The MAbs were of the isotypes IgG1, IgG2b and IgM. The MAbs reacted with VP28 of WSSV and not with any other viral or shrimp protein in western blot. The MAbs were used to develop dot immunoblot assay using an immunocomb to detect WSSV from field samples. The test developed had an analytical sensitivity of 625 pg and a diagnostic sensitivity of 100% compared to single step polymerase chain reaction (PCR). The test can be used as an alternate for first step PCR to detect WSSV from field samples.  相似文献   

2.
White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost‐effective strategy for long‐term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 103 fold higher in disease susceptible than that in disease resistant shrimps using real‐time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease‐free shrimp aquaculture industry.  相似文献   

3.
Hematological and Immunological parameters of aquatic animals may be changes due to acute, subacute and chronic exposure to marine pollutants. The purpose of this study was to determine the experimental effects of water-soluble fraction (WSD) doses (0, 10, 100, 500 and 1,000?ppm) for 0, 48?h and 7?days on hematological and immunological features of juvenile great sturgeon Huso huso. Fish exposed after 48?h and 7?days showed a significant change in the white blood cell (P?<?0.01) in contrast to red blood cell (P?>?0.05). Only MCV, neutrophil and lymphocyte show significant change within 48-h exposure to WSD (P?<?0.05), whereas among significant indices, MCV and lymphocyte had been decreased and neutrophil had increased. Seven-day exposures showed a significant change in MCV, neutrophil, eosinophil and lymphocyte concentration in relation to the respective control (P?<?0.05), whereas among significant indices, neutrophils were significantly greater and MCV, eosinophil and lymphocyte were significantly lower than those in control groups (P?<?0.05).  相似文献   

4.
Weissella ceti is an emerging bacterial pathogen that affects rainbow trout, Oncorhynchus mykiss (Walbaum), farms. The aims of this study were to genotype W. ceti strains isolated from distinct geographical origins and to determine the efficacy of an oil-adjuvanted vaccine against the disease. Between 2010 and 2012, outbreaks were recorded in five Brazilian farms, and 34 W. ceti isolates were genetically characterized by repetitive extragenic palindromic PCR, enterobacterial repetitive intergenic consensus sequences PCR and pulsed-field gel electrophoresis. Two different W. ceti vaccines were tested: an aqueous-based whole-cell inactivated vaccine (bacterin) and oil-adjuvanted vaccine. Their efficacy was evaluated in rainbow trout at 30 and 60 days post-vaccination (d.p.v.). W. ceti was found to be a highly homogeneous population in Brazil, with clonally related genotypes. Oil-adjuvanted vaccine exhibited the best (P < 0.05) protection against disease, reaching relative percentage survival (RPS)values of 92% at 30 and 60 d.p.v. Bacterin resulted in RPS values of 67% and 58% at day 30 and 60, respectively. The oil-adjuvanted vaccine provided effective protection against W. ceti infection in rainbow trout.  相似文献   

5.
To reduce disease impact, Philippine farmers developed the green-water (GW) system, which has been spreading rapidly since 2008. In the most applied GW-system, the shrimp pond receives water from a reservoir stocked with tilapia. We collected financial data on GW and non-GW systems from farms having similar management and environmental context. All farms had more than one pond; the average pond area was < 1 ha, and total pond areas per farm ranged from 1.4 to 139 ha. The total variable and fixed costs per ha of shrimp pond were not different for GW and non-GW farms, but on GW farms the cost per kg of shrimp produced was lower. The latter was higher than the 2009 market price of 30 g shrimp for non-GW farms mainly. In GW farms the cost of seed was lower, and survival rates and individual shrimp weights at harvest were higher for an equally long culture period.  相似文献   

6.
A new skin condition, known as puffy skin disease (PSD), emerged in farmed rainbow trout Oncorhynchus mykiss (Walbaum) in 2002. The number of new cases increased considerably from 2006. Clinical signs include white or grey skin patches, which become raised and red with excessive mucous production and scale loss. Fish are inappetant and lose condition. Histologically, the key feature is epithelial hyperplasia. We undertook a questionnaire study of trout farmers in England and Wales to investigate prevalence and risk factors. PSD was reported on 37% (n = 49) of rainbow trout sites, located in 28 river catchments. The increase in cases from 2006 onwards was mirrored by the increase in red mark syndrome (RMS). Prevalence and severity of PSD were highest in the summer months. The presence of PSD was associated with RMS (OR = 9.7, P < 0.001). Sites receiving live rainbow trout in the previous 12 months were considerably more likely to have PSD (OR = 5.3. P < 0.01), which suggests an infectious aetiology. The size of affected fish and prevalence varied between farms, indicating that farm‐level factors are important. Future research should further investigate the aetiology of PSD and practices to manage the disease.  相似文献   

7.
Infectious haematopoietic necrosis virus (IHNV) is the causative agent of infectious haematopoietic necrosis, a disease of salmonid responsible for great economic losses. The disease occurs in most parts of the world where rainbow trout is reared but has not been previously reported in Kenya. In this study, rainbow trout fry and growers from two farms in Nyeri County were screened for IHNV. Whole fry (n = 4 from each farm) and kidney samples from growers (n = 15 and n = 6 from the two farms, respectively) were collected and preserved for cell culture examination or PCR analysis. Screening of samples was done by PCR followed by sequencing of the glycoprotein gene of the virus. Demonstration of the virus was done by propagation in EPC cells followed by the indirect fluorescence antibody test (IFAT). The results revealed the presence of IHNV at low prevalence of 0.1 and 0.4 for the two farms. The virus was confirmed both by IFAT and by partial sequencing of the G gene. Phylogenetic analysis revealed that the Kenyan isolates were identical to those of the J genogroup found mostly in Asia. The findings have implications for biosecurity measures and import regulations for the Kenyan rainbow trout industry.  相似文献   

8.
White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV‐positive farms: post‐larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7–100% nucleotide sequence identity with four strains in the GenBank database – China ( AF332093 ), Taiwan ( AF440570 and U50923 ) and Thailand ( AF369029 ). This is the first broad study of WSSV infection in L. vannamei in Taiwan.  相似文献   

9.
In recent years, the shrimp industry has turned to inland freshwater culture as one method to avoid problems such as the introduction of possible vectors of viral pathogens into seawater ponds. Our experiments evaluated susceptibility to white spot syndrome virus (WSSV) in Litopenaeus vannamei held under different salinity regimens. Juvenile L. vannamei that were conditioned at salinities of 35, 25, 15, 5 and 2 g L−1 were challenged with WSSV. In order to assess the severity of white spot disease, histological analysis and nested polymerase chain reaction (PCR) tests were carried out on the challenged shrimp every 4 h after 48 h post challenge. The results indicated that significantly more severe infections resulted at 15‰ than at other salinities. Mortality could not be compared due to the sampling design and because severe WSSV infections occurred in all test groups such that few shrimp remained alive in each challenged group at the end of the test. Despite this, the results suggest that salinity may affect the course and outcome of WSSV infections.  相似文献   

10.
Integrated multi‐trophic aquaculture (IMTA) is an alternative approach to mono‐culture aquaculture that reduces environmental impacts of commercial aquaculture systems by combining the cultivation of fed species with extractive species. Shellfish play a critical role in IMTA systems by filter‐feeding particulate‐bound organic nutrients. They may also increase or decrease disease risk on farms by serving as reservoirs or barriers for important finfish pathogens such as infectious salmon anaemia virus (ISAV). This study aimed to optimize culture and molecular assays in shellfish tissues and to determine the fate of ISAV in mussels, Mytilus edulis. To determine detection limits, qRT‐PCR and culture assays in both CHSE‐ and ASK cells were optimized in ISAV‐inoculated mussel tissue homogenates. Both qRT‐PCR and culture assays performed in ASK cells had comparable detection limits of 102.8 TCID50 mL?1. The ISAV RNA genome was consistently detected in digestive gland tissue of ISAV‐exposed mussels. Viable ISAV was not detected in mussel tissues by culture analysis in CHSE‐ and ASK cells. The fact that qRT‐PCR analysis resulted in positive cycle threshold (CT) values that corresponded to the detectable range of ISAV in ASK culture assays suggests that little to no viable ISAV particles are present in the mussel tissues.  相似文献   

11.
White leg shrimp, Penaeus vannamei, were collected on a monthly basis from grow‐out ponds located at Tamil Nadu and Andhra Pradesh states along the east coast of India for screening of viral and other pathogens. Totally 240 shrimp samples randomly collected from 92 farms were screened for white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV), infectious myonecrosis virus (IMNV) and Enterocytozoon hepatopenaei (EHP). The number of shrimp collected from shrimp farms ranged from 6 to 20 based on the body weight of the shrimp. All the shrimp collected from one farm were pooled together for screening for pathogens by PCR assay. Among the samples screened, 28 samples were WSSV‐positive, one positive for IHHNV and 30 samples positive for EHP. Among the positive samples, four samples were found to be positive for both WSSV and EHP, which indicated that the shrimp had multiple infections with WSSV and EHP. This is the first report on the occurrence of multiple infections caused by WSSV and EHP. Multiplex PCR (m‐PCR) protocol was standardized to detect both pathogens simultaneously in single reaction instead of carrying out separate PCR for both pathogens. Using m‐PCR assay, naturally infected shrimp samples collected from field showed two prominent bands of 615 and 510 bp for WSSV and EHP, respectively.  相似文献   

12.
Kudoa thyrsites (Myxozoa) encysts within myocytes of a variety of fishes. While infected fish appear unharmed, parasite‐derived enzymes degrade the flesh post‐mortem. In regions of British Columbia (BC), Canada, up to 4–7% of fillets can be affected, thus having economic consequences and impacting the competitiveness of BC's farms. K. thyrsites was monitored in two farms having high (HP) or low (LP) historical infection prevalence. At each farm, 30 fish were sampled monthly for blood and muscle during the first year followed by nine samplings during year two. Prevalence and intensity were measured by PCR and histology of muscle samples. In parallel, fillet tests were used to quantify myoliquefaction. Infections were detected by PCR after 355 and 509 degree days at LP and HP farms, respectively. Prevalence reached 100% at the HP farm by 2265 degree days and declined during the second year, whereas it plateaued near 50% at the LP farm. Infection intensities decreased after 1 year at both farms. Blood was PCR‐positive at both farms between 778 and 1113 degree days and again after 2000 degree days. This is the first monitoring project in a production environment and compares data between farms with different prevalence.  相似文献   

13.
Abstract. A disease survey was conducted in North East Sumatera, Indonesia, during June and July 1991. The objeet was to obtain information regarding the diseases present in shrimp culture faeilities. Shrimp were sampled for histopathological examination and details of the farm management and environment were recorded. The stocks sampled included some reported to be suffering from disease problems and some reported to be in good health. Samples were taken from 24 pond farms, five hatcheries and a broodstock fisherman. All farms and hatcheries cultured Penaeus monodon Fabricius, one farm also cultured Penaeus merguiensis de Mann, and two cultured Metapenaeus spp. Several wild caught P. monodon female broodstoek and some wild rice shrimp ( Acetes spp.) were also sampled. The disease conditions detected in P. monodon were monodon baculovirus (MBV), hepatopancreatic parvo-likc virus (HPV), septic hcpatopancreatic necrosis (SHN), bacterial septicaemia (BS), haernocytie enteritis (HE), lymphoid organ pathology (LOP), external fouling organisms (EFO), and a single unconfirmed case of infectious hypodcrmal and haemopoietic necrosis (IHHN). The Acetes spp. from one site had a microsporidean infection in the striated muscle of the abdomen. EFO was the most common disease condition, and was identified in 16 farms and three hatcheries. MBV was detected in 14 farms, three hatcheries and in one broodstoek. SHN was found in 13 farms with one case of concurrent BS and LOP was found in eight farms. HE was found in four farms and HPV was identified in shrimp with MBV on two farms and as the only disease in one broodstoek. The significance and implications of these findings for the developing industry in Indonesia arc discussed.  相似文献   

14.
Tilapia lake virus disease (TiLVD) has emerged to be an important viral disease of farmed Nile tilapia (Oreochromis niloticus) having the potential to impede expansion of aquaculture production. There is a need for rapid diagnostic tools to identify infected fish to limit the spread in individual farms. We report the first detection of TiLV infection by PCR in farmed and wild Nile tilapia from Lake Victoria. There was no difference in prevalence between farmed and wild fish samples (p = .65), and of the 442 samples examined from 191 fish, 28 were positive for TiLV by PCR. In terms of tissue distribution, the head kidney (7.69%, N = 65) and spleen (10.99%, N = 191), samples had the highest prevalence (p < .0028) followed by heart samples (3.45%, N = 29). Conversely, the prevalence was low in the liver (0.71%, N = 140) and absent in brain samples (0.0%, N = 17), which have previously been shown to be target organs during acute infections. Phylogenetic analysis showed homology between our sequences and those from recent outbreaks in Israel and Thailand. Given that these findings were based on nucleic acid detection by PCR, future studies should seek to isolate the virus from fish in Lake Victoria and show its ability to cause disease and virulence in susceptible fish.  相似文献   

15.
Water as a habitat is highly variable in terms of the physico‐chemical properties. The aim of the studies was to analyze the quality of the technology by evaluating the non‐specific cellular defence potential of rainbow trout (Oncorhynchus mykiss) in diversified intensity culture systems and in different periods of the breeding cycle. The evaluation was conducted in order to establish the scientific basis for the development of principles for recognition of the culture of this species as ‘organic’. Six rainbow trout rearing farms were selected for the studies and divided into two equal groups according to the production technology: farms with a flow through system (with single water usage – OOH) and facilities with multiple water usage via its backflow (re‐circulating system – RAS). The samples for tests were taken from 20 individuals from each fish farm. In all fish that originated both from OOH‐type farms and RAS operations, an increase in the activity of non‐specific cellular immunity in the autumn was observed. Statistically significant reduction in the activity of cellular defensive mechanisms in small (300–500 g) and big (501–850 g) fish occurred on two 3‐OOH and 3‐RAS farms in spring and autumn in 2011. During these periods, the research has shown the presence of Infectious Pancreatic Necrosis Virus (IPNV), which provoked an immune suppression. Based on the obtained results it is concluded that there were no significant differences in the parameters of non‐specific cellular immunity in rainbow trout originating from different farming systems OOH and RAS.  相似文献   

16.
The culture of the Eastern oyster (Crassostrea virginica) in containment gear has become a viable component of restoration programs in many states on the East Coast of the United States, and it has been suggested that these operations may provide many of the same ecological services as natural or restored reefs. Our two-part study comparing the diversity and abundance in macro-epifaunal communities associated with a sub-tidal-created oyster reef and ‘modified rack and bag’ cage system (Part I) and floating oyster cages for restoration (Part II) occurred over the summer and fall of 2006 and 2007, respectively. In Part I, a greater total abundance and species richness (P < 0.05) was found to be associated with the cages, but greater evenness (P < 0.05) was found on the reef. No significant difference (P > 0.05) was found in species diversity according to Simpson’s index by habitat type, but it was significant (P < 0.05) by month. These samples were dominated by naked goby (Gobisoma bosc) and Atlantic mud crab (Panopeus herbstii). Spaghetti worm (Ampharetidae, P < 0.01), sheepshead (Archosargus probatocephalus, P < 0.01), blue crab (Callinectes sapidus, P < 0.01), grey snapper (Lutjanus griseus, P < 0.05), gag grouper (Mycteroperca microlepis, P < 0.01), and Atlantic oyster drill (Urosalpinx cinera, P < 0.05) were unique to the cages, while the skilletfish (Gobiesox strumosus) was unique to the reef. Part II revealed that the floating cages supported 13 species of fish and invertebrates, although no significant differences in species richness, evenness, or diversity were found by month or by bay area (P > 0.05). These results suggest that created reefs in conjunction with ‘rack and bag’ cage systems, as well as floating cage systems, support ecologically and economically important macro-epifauna, even at very small scales.  相似文献   

17.
Streptococcus dysgalactiae is an emerging fish pathogen that is responsible for outbreaks of disease on fish farms around the world. Recently, this bacterium was associated with an outbreak at a Nile tilapia, Oreochromis niloticus (L.), farm in Brazil. The aim of this study was to evaluate the genetic diversity, best genotyping method and aspects of molecular epidemiology of S. dysgalactiae infections in Nile tilapia farms in Brazil. Twenty‐one isolates from four farms located in different Brazilian states were characterized genetically using pulsed‐field gel electrophoresis (PFGE), ERIC‐PCR, REP‐PCR and sodA gene sequencing. The discriminatory power of the different typing methods was compared using Simpson's index of diversity. Identical sodA gene sequences were obtained from all isolates, and ERIC‐PCR and REP‐PCR were unable to discriminate among the isolates. PFGE typing detected three different genetic patterns between the 21 strains evaluated; thus, it was the best genotyping method for use with this pathogen. The strains from Ceará State were genetically divergent from those from Alagoas State. The S. dysgalactiae isolates analysed in this study constituted a genetically diverse population with a clear association between geographical origin and genotype.  相似文献   

18.
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19.
The objective quality indices of squid (Photololigo duvaucelii) and cuttlefish (Sepia aculeata) stored in ice were compared with the subjective counterparts. Sensory (overall quality rating, quality index method [QIM], and multisample difference test), microbiological (total viable count [TVC], psychrophilic count), chemical (trichloroacetic acid-soluble peptide [TCA-soluble peptide], trimethylamine nitrogen [TMA-N], total volatile bases nitrogen [TVB-N], ammonia content, and protein pattern), and physical analyses (expressible drip, color, and texture) were determined in both species during 16 days of iced storage. As storage time increased, TCA-soluble peptide, TVB, ammonia content, and expressible drip were increased (p < 0.05). TMA content was markedly increased after 10 and 8 days of storage in squid and cuttlefish, respectively. Both TVC and psychrophilic count increased as the storage time increased (p < 0.01). Myosin heavy chain was degraded with coincidental decrease in shear force and sensory texture during storage (p < 0.05). According to the overall rating score, shelf life of both species in ice was estimated to be 6 days. The increases in ammonia content and expressible drip were highly correlated with the decrease in overall quality rating and increase in quality index score of squid and cuttlefish (p < 0.01).  相似文献   

20.
The effect of the inclusion of probiotics (Vibrio alginolyticus) and β-1,3/1,6-glucans in Penaeus vannamei larviculture was evaluated by measuring the immune response and survival of shrimp juveniles subjected to white spot syndrome virus (WSSV) challenge tests (per os) and pond culture. Treatments were designed to contrast the probiotic factor (inclusion vs non-inclusion) and β-1,3/1,6-glucans supplementation in various larvae stages; starting from early stage (Zoea II), to middle stage (PL 12), late stage (15 days pre-challenge) or non-supplemented. In larviculture, the highest survival was obtained in animals treated with β-1,3/1,6-glucans from Zoea II. The use of probiotics enhanced survival during the first 0-52 h post-WSSV challenge period. During 56-156 h post-WSSV challenge period, interactions were observed between β-1,3/1,6-glucans × time and β-1,3/1,6-glucans × time × probiotics. Significant differences in final survival rates between treatments were not observed. In the second WSSV challenge, immune parameters were analysed. Significant interaction between probiotics and β-1,3/1,6-glucans was observed for plasmatic protein (PP) concentration, super oxide anion (O2−) generation, antibacterial activity (AA), and total haemocyte count (THC). The use of probiotics in larviculture had a negative effect on the PP, but increased the AA and THC, while β-1,3/1,6-glucans had a negative effect on the O2− generation. The most relevant results were obtained from treatments T2 (probiotics in larviculture, β-1,3/1,6-glucans from Z II) and T4 (probiotics in larviculture, β-1,3/1,6-glucans 15 days before challenge). Treatment T2 presented the highest survival rate in larviculture. After WSSV infection, the animals of this treatment displayed resistance to the virus, a strong AA and increase of THC. Treatment T4 increased the amount of PP, increased the O2− generation and THC. Histological analysis showed that the animals of treatment T2 and T4 were able to limit the spread of the virus during the first hour after challenge with WSSV. The survivors from treatments T2 and T4 had a high THC, accompanied by a lack of white spot disease (WSD) injuries. A bioassay was carried out under farm conditions during the warm-rainy season using larvae from treatments T2 and T4. The animals were stocked at 18 animals/m2 in earth ponds of 0.20 ha (three ponds/larvae kind). WSD outbreak was not detected, and the survival was significantly higher in ponds stocked with larvae from treatment T4 (70 ± 3%) than in ponds stocked with larvae from treatment T2 (49 ± 9%).  相似文献   

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