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The effect of plants and probiotics on the survival and immune response of Litopenaeus vannamei challenged with the white spot syndrome virus (WSSV) was evaluated. A probiotic mixture (PM), plant extract (PE) or powdered plants (PP) were added to feed with the attractant Dry Oil®. An experiment was conducted with five treatments in triplicate. Shrimp (weighing 11.70±2.5 g) were cultured in 120 L plastic tanks and fed twice a day with commercial feed plus additives or with commercial feed plus WSSV. Animals were monitored for the occurrence of WSSV using single‐step and nested PCR. The PM and PP added to the commercial feed showed high survival, a decrease in WSSV prevalence in shrimp and an increase in the activity of lysosomal enzymes, N‐acetyl‐β‐glucosaminidase and acid phosphatase. The total haemocyte count in shrimp treated with PM was significantly higher than that in the control group (treatment I) and in shrimp fed with PE. The results of the present work indicate that PP and PM are good candidates for use as feed additives against WSSV in shrimp cultures.  相似文献   

3.
In this study, we tested the lower salinity tolerance of juvenile shrimps (Litopenaeus vannamei) at a relatively low temperature (20 °C). In the first of two laboratory experiments, we first abruptly transferred shrimps (6.91 ± 0.05 g wet weight, mean ± SE) from the rearing salinity (35 000 mg L?1) to salinities of 5000, 15 000, 25 000, 35 000 (control) and 40 000 mg L?1 at 20 °C. The survival of L. vannamei juvenile was not affected by salinities from 15 000 to 40 000 mg L?1 during the 96‐h exposure periods. Shrimps exposed to 5000 mg L?1 were significantly affected by salinity, with a survival of 12.5% after 96 h. The 24‐, 48‐ and 96‐h lethal salinity for 50% (LS50) were 7020, 8510 and 9540 mg L?1 respectively. In the second experiment, shrimps (5.47 ± 0.09 g wet weight, mean ± SE) were acclimatized to the different salinity levels (5000, 15 000, 25 000, 35 000 and 40 000 mg L?1) and then maintained for 30 days at 20 °C. Results showed that the survival was significantly lower at 5000 mg L?1 than at other salinity levels, but the final wet weight under 5000 mg L?1 treatment was significantly higher than those under other treatments (P<0.05). Feed intake (FI) of shrimp under 5000 mg L?1 was significantly lower than those of shrimp under 150 00–40 000 mg L?1; food conversion efficiency (FCE), however, showed a contrasting change (P<0.05). Furthermore, salinity significantly influenced the oxygen consumption rates, ammonia‐N excretion rates and the O/N ratio of test shrimps (P<0.05). The results obtained in our work provide evidence that L. vannamei juveniles have limited capacity to tolerate salinities <10 000 mg L?1 at a relatively low temperature (20 °C). Results also show that L. vannamei juvenile can recover from the abrupt salinity change between 15 000 and 40 000 mg L?1 within 24 h.  相似文献   

4.
Growth, immunological and physiological parameters of white shrimp Litopenaeus vannamei reared at different salinity levels (1, 10, 15, 25 and 35 g/L) at stocking density of 214 shrimp/m3 were examined at 1, 30 and 63 days. Results showed that the total haemocyte count (THC) of shrimp decreased with time at all salinity levels, indicating a potential reduction in the resistance of shrimp against pathogens, since a low value of THC indicates a perturbation of the immune system. Glucose and protein values observed in the haemolymph throughout the study indicate that shrimp adapted well to low salinities (1, 10 and 15 g/L). Although of those shrimp reared at 10 g/L only 83.3% survived, at this salinity, shrimp depicted a higher glucose concentration in haemolymph at the beginning and end of the study.  相似文献   

5.
为探讨转vp28蓝藻(Anabaena sp.PCC7120)口服剂对凡纳滨对虾抗白斑综合征病毒能力及其相应的免疫反应,本研究将此口服剂免疫幼虾7 d,再分别通过投喂攻毒和浸泡攻毒,测定其存活率及相应的免疫指标。投喂攻毒和浸泡攻毒的实验组存活率分别为78.8%和83.19%,表明该口服剂能显著增强对虾抗白斑综合征病毒的能力。蓝藻口服剂免疫对虾的酶活性检测结果显示,超氧化物歧化酶(SOD)、酚氧化酶(PO)、过氧化氢酶(CAT)和碱性磷酸酶(AKP)活性在免疫后2 h均有上升趋势,且在48或96 h达到最高值,这表明该口服剂能引起对虾体内酶活性变化。投喂攻毒的对虾酶活性检测结果显示,实验组攻毒后的对虾肝胰腺SOD活性分别比阳性对照组、野生型组、空载体组显著提高42.10%、32.26%和16.04%,且攻毒后的肌肉SOD活性分别比阴性对照组、阳性对照组、野生型组和空载体组略微提高17.70%、11.50%、15.00%以及10.00%。实验组攻毒后的对虾肝胰腺PO、CAT和AKP活性比阳性对照组分别提高12.17%、88.80%和240.07%,比野生型组分别提高21.49%、30.90%和100%;酸性磷酸酶(ACP)活性比阴性对照组略微提高,而在肌肉中各组ACP活性无显著性差异。同时浸泡攻毒组结果与投喂攻毒组具有类似的趋势。浸泡攻毒的实验组CAT和AKP活性显著高于其余处理组,且CAT活性比投喂攻毒更为显著。浸泡攻毒的实验组肝胰腺PO活性显著高于阳性对照组、野生型组和空载体组,而各组肌肉ACP活性无显著性差异。研究表明,转vp28蓝藻口服剂能够增强凡纳滨对虾抗病能力并延缓对虾死亡。转vp28蓝藻PCC7120本身可作为幼虾饵料直接投喂,无需提取纯化,有望大规模应用于对虾养殖产业。  相似文献   

6.
In the field, moulting and salinity drop in the water due to excessive rainfall have been mentioned to be risk factors for WSSV outbreaks. Therefore, in this study, the effect of an acute change in environmental salinity and shedding of the old cuticle shell on the susceptibility of Penaeus vannamei to WSSV was evaluated by immersion challenge. For testing the effect of abrupt salinity stress, early premoult shrimp that were acclimated to 35 g L?1 were subjected to salinities of 50 g L?1, 35 g L?1, 20 g L?1, 10 g L?1 and 7 g L?1 or 5 g L?1 and simultaneously exposed to 105.5 SID50 mL?1 of WSSV for 5 h, after which the salinity was brought back to 35 g L?1. Shrimp that were transferred from 35 g L?1 to 50 g L?1, 35 g L?1 and 20 g L?1 did not become infected with WSSV. Shrimp became infected with WSSV after an acute salinity drop from 35 g L?1 to 10 g L?1 and lower. The mortality in shrimp, subjected to a salinity change to 10 g L?1, 7 g L?1 and 5 g L?1, was 6.7%, 46.7% and 53.3%, respectively (P < 0.05). For testing the effect of moulting, shrimp in early premoult, moulting and post‐moult were immersed in sea water containing 105.5 SID50 mL?1 of WSSV. The resulting mortality due to WSSV infection in shrimp inoculated during early premoult (0%), ecdysis (53.3%) and post‐moult (26.72%) demonstrated that a significant difference exists in susceptibility of shrimp during the short moulting process (P < 0.05). The findings of this study indicate that during a drop in environmental salinity lower than 10 g L?1 and ecdysis, shrimp are at risk for a WSSV infection. These findings have important implications for WSSV control measures.  相似文献   

7.
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   

8.
White spot syndrome virus (WSSV) is a major threat for farmed shrimp worldwide. RNA interference (RNAi) is the most recent tool against viral diseases. Rab7 silencing effectively inhibited virus infections in juvenile shrimp, but the antiviral effect in brooders remains unknown. This study found a homologue Penaeus monodon Rab7 gene in Litopenaeus vannamei brooders from Mexico. Sequence identity was >99% to a Thai LvRab7 sequence and >94% to Rab7 sequences from P. monodon or Marsupenaeus japonicus. Animals treated with a partial (494 bp) or a complete (618 bp) LvRab7 dsRNA sequences and challenged 48 h post treatment (hpt) with a high WSSV dose showed 80–88% mortality respectively. Shrimp treated with 4 or 20 μg LvRab7 dsRNA and challenged with a WSSV high dose had 80% mortality each, but it was reduced to 33% and 40%, respectively, with a low dose. Efficacy of dsRNA to reduce shrimp mortality was dependent on virus dose used regardless of dsRNA concentration. A significant reduction in LvRab7 mRNA levels was observed at 120 hpt. In conclusion, silencing LvRab7 in brooders showed a mild antiviral effect against a WSSV challenge at 48 hpt.  相似文献   

9.
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   

10.
Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) are two widespread shrimp viruses. The interference of IHHNV on WSSV was the first reported case of viral interference that involved crustacean viruses and has been subsequently confirmed. However, the mechanisms underlying the induction of WSSV resistance through IHHNV infection are practically unknown. In this study, the interference mechanisms between IHHNV and WSSV were studied using a competitive ELISA. The binding of WSSV and IHHNV to cellular membrane of Litopenaeus vannamei was examined. The results suggested that there existed a mutual competition between IHHNV and WSSV for binding to receptors present on cellular membrane of L. vannamei and that the inhibitory effects of WSSV towards IHHNV were more distinct than those of IHHNV towards WSSV.  相似文献   

11.
Dendronereis spp. (Peters) (Nereididae) is a common polychaete in shrimp ponds built on intertidal land and is natural food for shrimp in traditionally managed ponds in Indonesia. White spot syndrome virus (WSSV), an important viral pathogen of the shrimp, can replicate in this polychaete (Desrina et al. 2013); therefore, it is a potential propagative vector for virus transmission. The major aim of this study was to determine whether WSSV can be transmitted from naturally infected Dendronereis spp. to specific pathogen‐free (SPF) Pacific white shrimp Litopenaeus vannamei (Boone) through feeding. WSSV was detected in naturally infected Dendronereis spp. and Penaeus monodon Fabricius from a traditional shrimp pond, and the positive animals were used in the current experiment. WSSV‐infected Dendronereis spp. and P. monodon in a pond had a point prevalence of 90% and 80%, respectively, as measured by PCR. WSSV was detected in the head, gills, blood and mid‐body of Dendronereis spp. WSSV from naturally infected Dendronereis spp was transmitted to SPF L. vannamei and subsequently from this shrimp to new naïve‐SPF L. vannamei to cause transient infection. Our findings support the contention that Dendronereis spp, upon feeding, can be a source of WSSV infection of shrimp in ponds.  相似文献   

12.
White spot syndrome virus has been a threat to the global shrimp industry since it was discovered in Taiwan in 1992. Thus, shrimp-producing countries have launched regulations to prevent import of WSSV-infected commodity shrimp from endemic areas. Recently, cooked shrimp that is infected with WSSV tested positive by PCR. However, there is no study to determine the infectivity of WSSV in cooked shrimp that tested positive by PCR. In the present study, WSSV-infected shrimp were cooked at boiling temperature for different times including 0, 1, 3, 5, 10 and 30 min. Upon exposure to boiling temperature, WSSV-infected shrimp were fed to SPF shrimp (Litopenaeus vannamei). The result showed experimentally challenged shrimp from 0-min treatment (positive control) indeed got infected with WSSV. However, experimentally challenged shrimp that were fed tissues boiled at 1, 3, 5, 10 and 30 min were not infected with WSSV. Mortality data showed that only the positive control (0-min) treatment displayed high mortality, whereas no mortality was observed in any other treatment category. These findings suggest that cooking shrimp at boiling temperature for at least 1 min might prevent any potential spread of WSSV from endemic countries to other geographical areas where WSSV has not yet been reported.  相似文献   

13.
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   

14.
Litopenaeus vannamei (Boone) grown in ponds are exposed to salinities of less than 5 g L?1 during inland shrimp culture or to more than 40 g L?1 from evaporation and reduced water exchange in dry, hot climates. However, dietary requirements for shrimp grown in low or high salinities are not well defined, particularly for fatty acids. Feeding shrimp postlarvae with highly unsaturated fatty acids (HUFA) enhances tolerance to acute exposure to low salinity, as a result of better nutritional status, or/and specific effects of HUFA on membrane function and osmoregulation mechanisms. This study analysed the effect of HUFA supplementation (3% vs. 34%) on L. vannamei juveniles reared for 21 days at low (5 g L?1), medium (30 g L?1) and high salinities (50 g L?1). Juveniles grown at 5 g L?1 had lower survival compared with controls (30 g L?1) or shrimp grown at 50 g L?1, but no significant effect on survival was observed as a result of HUFA enrichment. In contrast, growth was significantly lower for shrimp grown at 50 g L?1, but this effect was compensated by the HUFA‐enriched diet. Osmotic pressure in haemolymph was affected by salinity, but not by HUFA enrichment. Shrimp fed HUFA‐enriched diets had significantly higher levels of eicosapentaenoic acid and docosahexaenoic acid in hepatopancreas and gills. These results demonstrate that growth at high salinities is enhanced with diets containing high HUFA levels, but that HUFA‐enriched diets have no effect on shrimp reared at low salinities.  相似文献   

15.
The white spot syndrome virus (WSSV) is a pathogen of great concern to the worldwide shrimp culture. In comparative studies on the WSSV genome, regions such as the open reading frame (ORF) 14–15 and ORF 23–24, prone to deletions and recombination, had been useful to study the evolutive relationships among viral strains. When looking for the WSSV strains infecting Litopenaeus vannamei (Boone) in northwest Mexico, we found evidence of a genetic similarity in ORF 14–15 to a strain from India and a recombination involving ORFs 78, 79 and 80. Two genotypes were found involving the insertion of a 265 base‐pair segment of ORF 108 into ORF 78 with inversions and deletions within ORFs 78, 79 and 80. The WSSV has an Asian origin and the mutations found could be an adaptation strategy to infect L. vannamei and other crustacean species of the American continent.  相似文献   

16.
Temperature and salinity are two factors known to influence the growth potential and survival of the Pacific white shrimp, Litopenaeus vannamei, acclimated to low salinity waters. In west Alabama, farmers suspect low water temperatures at stocking, in conjunction with low salinity and suboptimal ionic profiles, might be responsible for reduced survival and production at harvest. To determine the influence of temperature and salinity on post‐larval (PL) L. vannamei, a series of bioassays were conducted at the E.W. Shell Fisheries Research Station in Auburn, Alabama and Claude Peteet Mariculture Center in Gulf Shores, Alabama. PL L. vannamei of ages 11, 13, and 20 (PL11, PL13, and PL20) were acclimated down to salinities of 12, 4, 2, 1, 0.5, and 0.2 ppt at different temperatures ranging from 17.6 to 24.0 C. During the acclimation bioassays survivals were assessed at 24 and 48 h. PL survival of the three age groups examined were significantly reduced at salinities of 1, 0.5, and 0.2 ppt. These results correspond well to those reported at higher temperatures confirming that across the tested temperature range salinity endpoint was the driving factor in determining survival and that suboptimal temperatures had a minimal influence on survival.  相似文献   

17.
The white spot syndrome virus (WSSV) remains the most devastating viral pathogen of shrimp culture worldwide. Gene silencing by RNA interference (RNAi) using double stranded RNA (dsRNA) has been considered a powerful tool for conferring protection against WSSV when viral genes are silenced, as documented in several shrimp species. However, this effect is not long lasting. Our results provide the first evidence that long‐term silencing of the LvRab7 endogen produced antiviral effect against WSSV, which endured at least 21 d after dsRNA treatment (dat). Until now, the most efficient way to implement RNAi with dsRNA into the shrimp is by injection. Consequently, its application to broodstock in hatcheries is possible, minimizing the risk of vertical transmission of the virus. We show that the expression of Rab7 in hemocytes is lowest at 2 dat and finally recovers to basal status. In contrast, in gills and pleopods, gene expression silencing continued for at least 21 d. We challenged Litopenaeus vannamei broodstock with WSSV at 7, 14, or 21 dat reaching mortality rates of 0, 40, and 27%, respectively. In conclusion, the LvRab7 gene silencing is progressive and effective against WSSV. However, further studies are necessary to elucidate the functions of Rab7 in shrimp cells before applying this methodology at a commercial level.  相似文献   

18.
Three isonitrogenous diets containing 60 g kg–1, 90 g kg–1 or 120 g kg–1 lipid were formulated and fed to the Litopenaeus vannamei (2.00 ± 0.08 g) under two salinities (25 or 3 psu) in triplicate for 8 weeks. Shrimp fed 90 g kg–1 lipid had higher weight gain and specific growth rate than shrimp fed the other two diets regardless of salinity, and the hepatosomatic index increased with increasing dietary lipid at both salinities. The shrimp at 3 psu had significantly lower survival and ash content, higher condition factor, weight gain and specific growth rate than the shrimp at 25 psu. Increasing dietary lipid level induced the accumulation of serum MDA regardless of salinity, and at 3 psu, it reduced the serum GOT and GPT activities and the mRNA expression of TNF‐α in intestine and gill of L. vannamei. The hepatopancreatic triacylglycerol lipase (TGL) and CPT‐1 mRNA expression showed the highest value in shrimp fed 90 g kg–1 lipid diet at 3 psu. This study indicates that 120 g kg–1 dietary lipid may negatively affect the growth and induce oxidative damage in shrimp, but can improve immune defence at low salinity; 60 g kg–1 dietary lipid cannot afford the growth and either has no positive impact on the immunology for L. vannamei at 3 psu.  相似文献   

19.
Increasing numbers of mislabelled seafood products in the globalized market underline the need for approaches to identify the origin of these products. Pacific white shrimp cultured in different salinities (Litopenaeus vannamei) from 16 locations in China and USA were collected and differentiated based on δ13C and δ15N values. The δ13C and δ15N values of 16 commercial feeds used in shrimp culture were measured and effects of feed on C and N stable isotope composition in shrimp were investigated. Carbon and nitrogen isotopic analysis of the shrimp did not provide means to discriminate farmed shrimp from China and USA. However, the shrimp cultured in water with high salinity were enriched in 13C compared to shrimp cultured in freshwater and this provided another basis for discrimination. The correlation between δ13C in feed and shrimp was not significant, while the δ15N in feed and shrimp showed a trend of positive correlation.  相似文献   

20.
An experiment was conducted to evaluate the effect of a hot water extract of brown seaweeds Sargassum duplicatum and Sargassum wightii on the growth and white spot syndrome virus (WSSV) resistance in shrimp Penaeus monodon postlarvae (PL). Artemia nauplii (instar II) were enriched with both seaweed extracts at various concentrations (250, 500 and 750 mg L?1) and fed to the respective P. monodon (PL15–35) group for 20 days. A control group was also maintained without seaweed extract supplementation. The weight gain of the experimental groups was significantly higher (0.274–0.323 g) than the control group (0.261 g). Similarly, the specific growth rate was also significantly higher (16.27–17.06%) in the experimental groups than in the control group (16.03%). After 20 days of the feeding experiment, the shrimp PL were challenged with WSSV for 21 days. During the challenge test, the control shrimp displayed 100% mortality within 8 days. In contrast, the mortality percentage of the highest concentration (750 mg L?1) of seaweed extract enriched Artemia nauplii fed shrimp was 54–79%. Comparatively, low mortality was observed in S. wightii extract‐enriched Artemia nauplii fed shrimp. The polymerase chain reaction analysis indicated the concentration‐dependent infection of WSSV in P. monodon PL.  相似文献   

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