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Fish nodavirus (betanodavirus), a viral pathogen responsible for viral nervous necrosis (VNN) was isolated from infected Asian sea bass (Lates calcarifer). The distribution, clearance and expression of nodavirus vaccine, on the basis of DNA vaccine (pFNCPE42 DNA‐pcDNA3.1) construction, were analysed in tissues of the Asian seabass by PCR, RT‐PCR, ELISA and Immunohistochemistry. Fish immunized with a single intramuscular injection of 20 μg of the pFNCPE42‐DNA vaccine showed a significant increase in the serum antibody level in the 3rd week after vaccination, compared to control eukaryotic expression vector pcDNA3.1 vaccinated fish. Results from PCR studies indicated that the vaccine‐containing plasmids were distributed in heart, intestine, gill, muscle and liver 10 days after vaccination. Clearance of pFNCPE42‐DNA vaccine was studied at 10, 25, 50, 75 and 100 days of post vaccination (d p.v). At 100 days p.v. pFNCPE42‐DNA was cleared from muscle of vaccinated sea bass. In vitro and in vivo expression of fish nodavirus capsid protein gene (FNCP) was determined by fluorescent microscopy. Asian seabass was immunized with pFNCPE42‐DNA vaccine at a dose of 20 μg per fish and were challenged with betanodavirus by intramuscular injection. The vaccinated seabass was protected from nodaviral infection and 77.33% of relative percent survival (RPS) was recorded.  相似文献   

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To optimize broodstock management for Seriola rivoliana, a survey over 5 years was performed to evaluate the effects of successive hormonal inductions with GnRHa or dietary regime on spawning quality. Running males and females with an oocyte diameter > 500 μm were injected with GnRHa. The spawning quality was compared among consecutive years and different moments along the spawning season. Besides, three different feeding regimes were tested. Spawning quality parameters were not significantly affected by time along the spawning season, whereas they were clearly influenced by diet. Particularly, broodfish fed the mackerel regime (M) showed a significantly higher number of eggs than other treatments. Besides, dietary protein content significantly improved broodstock fecundity of S. rivoliana. Moreover, hatching rates were also higher in broodfish fed mackerel and the dietary docosahexaenoic acid (DHA) levels significantly increased this parameter. Despite polar lipids were not significantly affected, neutral lipids fatty acid composition of S. rivoliana eggs showed higher 18:2n‐6 in eggs, whereas ARA content was not affected. The results of this study denoted the high fecundity and good spawning quality of S. rivoliana broodfish and suggested the importance of high protein, energy and DHA levels in broodstock diets.  相似文献   

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The collection of fertilized eggs for mass culture of Pacific bluefin tuna (PBT) relies on the spontaneous spawning of broodstock in captivity, and the spawning season of the broodstock is generally from mid‐May to September. The diameter of fertilized eggs from the broodstock decreases during the spawning season. To investigate the influence of three potential factors, the egg diameter (larger or smaller), the rearing water temperature (25 or 28 C), and the aeration rate during the night (strong or weak), on early survival and on growth of PBT larvae, replicate trials were conducted to assess these three factors until 7 d after hatching (d.a.h.). At 7 d.a.h., survival rates of larvae reared with strong nighttime aeration were found by a three‐way ANOVA to be significantly higher than for larvae reared with weak aeration. Furthermore, growth rates of PBT larvae hatched from larger eggs were significantly faster than those from smaller eggs. However, a significant difference in the survival rate was not detected for the factors, egg diameter and rearing water temperature. The results indicate that rearing with strong nighttime aeration significantly improved the early survival of PBT larvae and the egg diameter (a proxy for egg quality) and influenced growth.  相似文献   

7.
Two experiments addressed the spontaneous spawning of red snapper, Lutjanus campechanus , under controlled temperatures and photoperiods and the effect of broodstock diets supplemented with oils rich in polyunsaturated fatty acids. In Experiment 1, broodfish were fed a standard diet (ST1) and one enriched with menhaden oil (ER1) over a 355-d period. ER1-influenced egg fatty acid profile, however, did not positively influence egg production. Both diets produced highly viable eggs and larvae but results varied within treatments. In Experiment 2, broodfish were fed either a standard diet (ST2) or one supplemented with oils (ER2) rich in docosahexaenoic and arachidonic acid using a 203-d cycle. Both treatments produced eggs but fertilization rates ranged 0–10%. There was no clear influence of the diets on egg fatty acid profiles. These results indicate that red snapper can spawn spontaneously in tanks under controlled environmental conditions and produce viable eggs and larvae when fed diets based on squid, shrimp, and fish. The fatty acid composition of the diets was reflected in the eggs to some degree, but the oil enrichments did not further enhance the reproductive performance and egg quality under the conditions of this study.  相似文献   

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It is hypothesized that the frequency of bacterial coldwater disease outbreaks can be reduced through the detection of the aetiologic agent, Flavobacterium psychrophilum, in broodstock followed by culling of eggs from heavily infected broodstock. Before a culling programme can be instituted, however, it is necessary to determine the sensitivity and specificity of existing assays for the detection of F. psychrophilum. In this study, tissue and ovarian fluid samples were collected from 224 fish at five hatcheries and screened using an enzyme‐linked immunosorbent assay (ELISA), a membrane‐filtration fluorescent antibody test (MF‐FAT), bacteriological culture and nested PCR. Latent class analysis was used to estimate sensitivity and specificity of kidney culture, kidney ELISA, nested PCR and MF‐FAT. Analytical sensitivity of the ELISA varied but was greatest when bacteria were cultured under iron‐limiting conditions. Diagnostic sensitivity estimates ranged from 0.02 (kidney culture) to 0.97 (kidney ELISA). Specificity estimates ranged from 0.02 (MF‐FAT) to 0.98 (kidney ELISA). In a separate challenge experiment, the ELISA confirmed the presence of F. psychrophilum in sub‐clinically infected fish. Results from this study demonstrate that the ELISA is an appropriate tool to screen broodstock and provides an indication of infection severity, which is crucial for implementation of a screening/culling programme.  相似文献   

9.
We previously established a method for spawning induction in Eastern little tuna (ELT) Euthynnus affinis (Cantor, 1849) by administering a gonadotropin‐releasing hormone analog (GnRHa) during the natural spawning season in Japan (August–October). In order to establish seed production of ELT in the off‐spawning season, we first conducted three spawning induction trials by GnRHa administration from October 2011 to January 2012 using ELT broodstock (2 years old; three females and four males) maintained in a 10‐m3 tank with a semi‐closed recirculation system and static elevated temperature. Average water temperature and daily egg production in three trials lasting 11–15 days were 27.0 ± 0.09°C and 268 173 eggs (Trial 1), 27.0 ± 0.11°C and 277 9098 eggs (Trial 2), and 25.5 ± 0.39°C and 291 113 eggs (Trial 3) respectively. Mean fertilization rate and mean hatching rate were 70.4% and 60.5% (Trial 1), 83.9% and 79.6% (Trial 2), and 62.5% and 57.4% (Trial 3) respectively. We also succeeded in producing ELT larvae in the pre‐spawning season (April–July), although the quantity and quality of larvae produced were inferior to those produced in other calendar months. In trials involving periodic GnRHa administration during the off‐spawning seasons, hatched larvae were obtained in the 10‐m3 tank after six of nine administrations in the 2011–2012 off‐spawning season and in 16 of 19 administrations in the 2012–2013 off‐spawning season. The findings of this study demonstrated that hormonal treatment and thermal control could be used to extend the spawning period in ELT, potentially allowing larval production in the post‐ and pre‐spawning seasons.  相似文献   

10.
Effects of dietary protein level and water salinity on spawning performance of Nile tilapia broodstock and growth of their larvae were studied. Four isocaloric (400 kcal/100 g) diets containing 25%, 30%, 35% and 40% crude protein were prepared. The diets were fed to broodfish (25.7 g) reared at three water salinities (0‰, 7‰ and 14‰) at a female/male ratio of 3:1, to satiation twice a day for 195 days. The size at first maturation increased with increasing dietary protein at all salinities. At 25% and 30% protein levels, broodstock reared at 0‰ reached their sexual maturity at bigger sizes than those reared at 7‰ and 14‰. At 0‰, spawning intervals were not significantly affected by dietary protein levels. At 7‰ and 14‰, spawning intervals significantly decreased with increasing dietary protein level. Spawning frequency and number of eggs per spawn were increased with increasing dietary protein level. The total number of spawnings per female and absolute fecundity were better in fish fed 40% protein in freshwater than at 7‰ and 14‰ salinity. The relationship of dietary protein and water salinity on egg size was significant, but showed irregular patterns. The chemical composition of broodstock muscles, eggs and fry were not significantly affected by dietary protein and water salinity, except for body water and crude protein of broodstock which were significantly affected; but showed irregular trends. At each water salinity, egg hatchability was linearly increased with increasing dietary protein level. Eggs produced from broodstock fed 25% protein at 7‰ and 14‰ needed more time for hatching and yolk-sac absorption and resulted in poorer larval weight than those reared in freshwater. Fry growth was improved with increasing protein level at all salinities. This result revealed that 40% dietary protein is required for optimum spawning performance of Nile tilapia reared at 0‰, 7‰ and 14‰ salinity. It also indicated that spawning performance and larval growth were better in freshwater than at 7‰ and 14‰.  相似文献   

11.
This study was conducted to investigate the effects of maternal inherited immunity acquired from crustacean‐enhanced diets on the vitality and profitability of sea bass offspring. Newly hatched larvae produced from three groups of broodstock were evaluated. The broodstock were fed (a) a basal diet (BD), (b) a Palaemon‐supplemented diet (PSD), and (c) an Artemia‐supplemented diet (ASD) for 42 days. A total of 400,000 larvae at 3 days posthatch (DPH) produced from each treatment were stocked in larval rearing tanks at 40 larvae/L for 42 days. Survival (%) was improved by 37 and 9.96% in the groups fed ASD and PSD compared with the control group. The growth, swim bladder (%), and condition factor all significantly (p ≤ 0.05) improved in the postlarvae produced from broodstock enhanced with crustacean diets. Compared with the BD group, the serum lysozyme activities of the fish groups fed ASD and PSD increased by 45.6 and 11.7%, respectively. Sea bass fry (90DPH) produced from broodstock fed ASD showed the best tolerance to salinity/temperature stress tests. Furthermore, the profitability improved in ASD and PSD compared with the BD group. In conclusion, sea bass broodstock enhanced with Artemia biomass produced offspring of superior quality with less cost and greater profit margins.  相似文献   

12.
Hormone‐induced spawning of southern flounder Paralichthys lethostigma has produced substantial numbers of viable eggs, but wide variations in fertilization and hatch rates have been reported. Recently, sustained natural spawning of southern flounder broodstock, without hormone induction, has been achieved in our laboratory. Adults (average weight = 1.12 kg; N= 25), including 6 captured as juveniles in 1993 and 19 captured as adults during September 1998, were stocked in two 4.8‐m3 controlled‐environment tanks in October 1998 and held under natural photothermal conditions until January 1999, when an artificial winter photo‐period of 10 L:14 D was initiated and then maintained through April 1999. Sex ratio was approximately 13 females:8 males:7 unknown. Natural spawning was observed in early December 1998 and increased in frequency to a peak in March 1999, before declining in late April. Water temperature ranged from 13.9 to 24.5 C during the spawning period. Natural spawnings over 142 d produced a total of 18.3 × 106 eggs, with a mean fertilization rate of 28.0% (range = 0–100%), yielding 4.94 × 106 fertilized eggs. The mean percentage of eggs that remained buoyant in full‐strength seawater (34 ppt) was 41.3% (0–98%), while hatching rate of buoyant eggs was 37.3% (0–99%) and survival of yolksac larvae to the first‐feeding stage was 30.2% (0–100%). Gonadal biopsies in late April identified six females from both tanks as probable spawners. A preliminary comparison suggests that natural spawning produced much larger numbers of viable eggs per female, with higher egg quality (i.e., fertilization and hatching success) than hormone‐induced spawning. In contrast to natural spawning, hormone‐induced strip‐spawning enabled timing of spawnings to be more precisely controlled. These results suggest that a combination of both natural and hormone‐induced spawning of photothermally conditioned fish will help produce the large numbers of eggs required to support commercial production.  相似文献   

13.
Losses due to cardiomyopathy syndrome (CMS) keep increasing in salmon‐producing countries in the North‐Atlantic. Recently, Piscine myocarditis virus (PMCV) has been detected in post‐smolts shortly after sea‐transfer, indicating a possible carry‐over from the hatcheries. In addition, there are reports of prevalences of PMCV as high as 70%–90% in certain groups of broodfish, and a recent outbreak of CMS in the Faroe Islands has been linked to the importation of eggs from a CMS‐endemic area. Thus, there is a need to investigate whether PMCV can be transmitted vertically from infected broodstock to their progeny. In the present study, samples from eggs, larvae, fingerlings and presmolt originating from PMCV‐positive broodstock from two commercial Atlantic salmon producers were tested for PMCV. The prevalence of PMCV in the broodstock was 98% in the hearts, 69% in the roe and 59% in the milt. Piscine myocarditis virus was detected in all stages of the progeny until and including the 40 g stage. Piscine myocarditis virus was also detected in presmolt sampled for tissue tropism. This provides farmers with several options for minimizing the risk of transfer of PMCV from broodstock to progeny, including screening of broodstock and aiming to use only those that are negative for PMCV or have low levels of virus.  相似文献   

14.
Commercial intensive fry production of Atlantic cod will be dependent on production of viable eggs independent of season. This can only be done by manipulation of maturation by photoperiod, but little is known about potential effects on egg characteristics and larval viability. In two cod broodstocks, maturation was successfully advanced or delayed 6 months compared with normal spawning season (March–April) by manipulation of photoperiod. The advanced broodstock spawned both in spring and autumn the same year. In two of the spawning tanks during autumn, ambient temperature was reduced after reaching 13.7°C during the first half of the spawning period. Egg quality and viability were monitored, and several egg batches were incubated, hatched and start‐fed for examination of growth and survival. Temperatures above 9.6°C resulted in significant reductions in fertilization and normal egg development. Concurrently, fractions of dead and unfertilized eggs increased with elevated temperature. Actual relative fecundity was not affected by temperature. Egg characteristics improved when temperature was controlled and lowered below 9.6°C. Occurrence of irregular spawners suggests that handling of broodstock fish should be avoided during maturation and spawning. Cod larvae originating from eggs of the advanced or delayed broodstocks were successfully reared beyond metamorphosis. Survival was 9.0–46.6% and 29.3% in green and clear water respectively. Survival correlated with both initial and average feeding conditions, but growth rate did not correlate with either of survival and feeding conditions. Specific growth rates (8.3–13.6% day−1) is comparable with other intensive rearing trials with cod, but were lower than reported from nature‐like systems.  相似文献   

15.
Eggs from a F1 cultured broodstock of sole were compared with eggs from wild‐caught breeders throughout one spawning season, to evaluate if egg quality may be affected by culture‐related conditions. Fourteen batches of eggs from cultured broodstock and 17 batches from wild‐caught sole were compared with respect to fatty acid (FA) composition, egg size, fertilization rate and hatching rate. Based on a multivariate analysis of the FA profiles, it was possible to discriminate between culture and wild inheritance. Eggs from cultured broodstock had high levels of C20:1(n‐9), C18:2(n‐6) and C18:3(n‐3), whereas eggs from wild fish had high levels of C16:1(n‐7), C20:4(n‐6) and C20:5(n‐3). Differences in FA profiles were most likely related to dietary differences. Fertilization and hatching rates were generally low and lowest in eggs from cultured broodstock, but not related to FA composition. Larval growth of one batch from each group was compared. Larval growth was not correlated to broodstock origin, FA composition or egg or larval size. However, larval survival was significantly lower for larvae from cultured broodstock.  相似文献   

16.
Viral nervous necrosis (VNN) affects more than 120 species mostly belonging to the order Perciformes. However, none of the brackishwater species belonging to the family Cichlidae under the order Perciformes are reported to be susceptible. Hence, the present experiment was undertaken to study the susceptibility of the brackishwater cichlid, pearlspot, Etroplus suratensis to NNV. Thirty‐day‐old pearlspot larvae were infected with NNV by immersion. Mortality was recorded till 14 days post‐infection, and the infected larvae were subjected to nested RT‐PCR and histology. The virus was isolated from infected larvae using SSN‐1 cells. To study the replication of the virus in vitro, primary cultured brain cells of E. suratensis and IEK cells were infected with NNV. No mortality was observed in any of the control or experimentally infected larvae. However, the experimentally infected larvae were positive for NNV by nested RT‐PCR and the virus was isolated using SSN‐1 cells. Further, the infected pearlspot brain cells and IEK cells showed cytopathic effect at second and third passage of the virus and they were positive for NNV by nested RT‐PCR. Pearlspot is relatively resistant to VNN although the virus could replicate in the larvae and in cell culture.  相似文献   

17.
The effects of four light intensities on growth and survival of first‐feeding stage black sea bass larvae Centropristis striata were investigated in a controlled‐environment laboratory. Fertilized eggs, obtained from LHRHa‐induced spawning of captive broodstock, were stocked (72 eggs L?1) into twenty 15 L black tanks under light intensities of 100, 500, 1000 and 1500 lx, with five replicate tanks per treatment. The photoperiod was 12L:12D, the temperature was 20°C and the salinity was 35 g L?1. Larvae were fed rotifers Brachionus rotundiformis from day 2 post‐hatching (d 2ph) at 5–10 rotifers mL?1. Microalgae Nannochloropis oculata and Isochrysis sp. were added (1:1) daily to maintain a density of 300 000 cells mL?1. Hatching success and larval growth and survival from d 2ph through d 15ph were monitored. Hatching success was 28–38% under all light intensities, and notochord length at hatching ranged from 2.8 to 3.0 mm, with no significant differences among treatments. By d 15ph, growth (mg wet weight) was significantly higher in the 1000 lx (0.914) and 1500 lx treatments (0.892) than in 100 lx (0.483), and a highly significant trend (P<0.01) towards increased survival with increasing light intensities was observed, from 1.3% at 100 lx to 13.9% at 1500 lx. Higher light intensities within the range of 100–1500 lx improved growth and survival of early larval black sea bass, suggesting that even higher light intensities may improve culture performance. This is consistent with conditions in shallow, near‐shore locations where eggs and larvae are distributed in nature.  相似文献   

18.
This study aims to evaluate the efficacy of Streptococcus agalactiae vaccine preparations, administered to tilapia broodstock, in preventing streptococcosis, through specific and non-specific immunity being transferred to the offspring. The study was conducted in two phases. The first was the vaccination of the broodstock using a whole-cell vaccine, an extracellular product (ECP) vaccine, and a combination of the two with a ratio of 1:1. The vaccines were administered to the broodstock 2 and 3 weeks before spawning. The second phase was the challenge test for larvae produced by vaccinated broodstock, and larvae from the unvaccinated control broodstock, through immersion in a suspension of 107 cfu mL?1 pathogenic S. agalactiae for 30 min, at ages 7, 14, 21, and 28 days post-hatching. The parameters evaluated were the broodstock’s blood profile, antibody-lysozyme (in broodstock, eggs, and larvae), and the larvae’s relative percent survival. Treatment with the combined vaccine administered 3 weeks before spawning resulted in the broodstock having significantly better antibody levels, lysozyme activity, and hematology profiles, compared to the other treatments (p?<?0.05). In addition, the larvae produced by broodstock subjected to this treatment, when challenged with the pathogenic S. agalactiae at ages 7, 14, 21, and 28 days, had RPS values of 95.24, 83.33, 72.22, and 56.02%, respectively. It was concluded that the administration of the “whole-cell/ECP” combination vaccine preparation to tilapia broodstock in the 3 weeks before spawning can increase specific and non-specific immunity in the broodstock and protect the larvae from S. agalactiae infection.  相似文献   

19.
The black sea bass is a high‐value marine serranid and is a prime candidate for intensive cultivation. Reliable methods for controlled spawning are needed to accelerate the development of hatchery technologies that result in mass production of healthy juveniles. During 1998–2001, spawning studies were conducted at The University of North Carolina at Wilmington (UNCW) and at the South Carolina Department of Natural Resources (SCDNR), Charleston, using pelleted luteinizing hormone releasing hormone analogue (LHRH‐a). From April through July 2001, 28 vitellogenic‐stage females, with mean oocyte diameters (MOD) ranging from 277–448 μm, were implanted with a 95% cholesterol‐5% cellulose pellet containing LHRH‐a (‐50 μg/kg body wt) at UNCW. In 10 individual spawning trials, females with MOD of 305–448 μm and maximum oocyte diameter × 475 μm spawned volitionally beginning 2–3 d post‐implantation (PI) and continued spawning over an average of 1.9 d (range = 1–4 d). Individual females released a mean total of 149,000 eggs (117,000 eggs/kg) with a mean buoyancy rate of 40.5% (floaters). Fertilization and hatching rates were 98% and 27.2% of floaters, respectively, yielding 14,600 yolksac larvae/female (12,600 yolksac larvae/kg body wt), and overall egg viability averaged 8.9%. In eight group spawning trials (2–3 females/group), average performance of females, including fecundity (103,800 eggs/female; 105,500 eggs/kg body wt), buoyancy rate (42.5%), fertilization and hatching rates (97.7% and 24.3% of floaters), numbers of yolksac larvae produced (10,900 yolksac larvae/female; 10,100 yolksac larvae/kg body wt), and overall egg viability (10.6%) was comparable to what was seen in individual spawning trials. From 1998–2000, a total of 58 vitellogenic stage (70% of oocytes 500 pm) females were implanted with pelleted LHRH‐a (‐50 μg/kg body wt) in nine group spawning trials (2–19 females/group) at SCDNR. Volitional spawning typically began 18–42 h PI and recurred every 1–3 d for an average duration of 9 d. Female groups released a mean of 560,000 eggs (84,000/female; 132,000/kg body wt) over the spawning period, with mean buoyancy rate of 25.7% floaters. Fertilization and hatching rates were 17.7% and 11.6 % of floaters, respectively, yielding 4,300 yolksac larvae/female (4,600 yolksac larvae/kg body wt). Overall egg viability was 2.9%. Captive wild‐caught black sea bass were induced to undergo repetitive volitional spawning by implantation of pelleted‐LHRH‐a, consistent with a multiple clutch group synchronous pattern of ovarian development. Group spawning appears to be a practical way to compensate for variable fecundity and egg viability of individual females. Research is needed to identify optimum hormone treatments and eligibility requirements.  相似文献   

20.
Successful natural spawning of Chaetodontoplus septentrionalis in captivity from 19 March to 11 May, 2008 is described for the first time. A single male dominates a harem of two females, spawning with each at dusk, from 10 min before to 20 min after sunset. Each female laid an average 119 × 103 eggs during the spawning period. Fertilized eggs were spherical, buoyant and had a diameter of 0.83 ± 0.02 mm (mean ± SD). Embryonic development lasted 15–18 h at 28.1 °C. Newly hatched larvae were 1.60 ± 0.07 mm in total length (TL) with 27 myomeres. Larvae completed yolk absorption within 3 days post hatching (ph) at 3.01 ± 0.08 mm TL. Ten days ph, the larvae had attained 3.95 ± 0.12 mm TL. Larvae were fed either 100% s‐type rotifers (Brachionus rotundiformis), 100% copepods (Microsetella sp.), a combination of the two (50%:50%) or without live feed (starved control) to determine the effect of live feed on the survival rate. The survival was significantly (P<0.001) higher in larvae fed a combination of diet than the others. These results indicate that C. septentrionalis is a potential species for captive breeding programs and the use of a combination of diet (s‐type rotifers and copepods) may be a suitable first food for the larvae.  相似文献   

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