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1.

The present study investigated the effects of monovalent and bivalent vaccines against Streptococcus iniae and Vibrio harveyi on growth performance, blood biochemical parameters, immune system, and antioxidant defense of Asian seabass (Lates calcarifer) fingerlings. Fish were exposed to seven experimental groups containing phosphate buffer saline (control), S. iniae vaccine by injection, S. iniae vaccine by immersion method, V. harveyi vaccine by injection, V. harveyi vaccine by immersion method, S. iniae/V. harveyi vaccines by immersion method, and S. iniae/V. harveyi vaccines by injection in triplicate for 60 days. Vaccination resulted in a significant alteration of the final weight (FW), feed conversion ratio (FCR), daily weight gain (DWG), relative growth rate (RGR), glucose, lysozyme, and complement levels at 60 days post-vaccination. Also, serum glutathione activity significantly increased in group SVIM (S. iniae/V. harveyi vaccines by immersion method) compared to the control group at 60 days post-vaccination. The level of MDA in serum was significantly decreased in SIN (S. iniae vaccine by injection) and SVIM groups compared to the control group 60 days post-vaccination where the value of this parameter was significantly higher in VIM (V. harveyi vaccine by immersion method) and SVIN (S. iniae/V. harveyi vaccines by injection) groups. The results of this study suggest that exposure to monovalent and bivalent vaccines against S. iniae and V. harveyi could alter the growth performance, nonspecific immune response, and antioxidant defense of Asian seabass fingerlings.

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2.
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   

3.
The aim of this study is the development and evaluation of a rapid and accurate quantitative PCR (qPCR)‐based protocol for detection of zoonotic pathogen Streptococcus iniae in bacterial cultures and tissues of diseased fish. For this purpose, the lactate permease‐encoding (lldY) gene was selected as a target for the design of S. iniae‐specific primers based on comparative genomic analysis using 45 sequences retrieved from NCBI genome database. Specificity and applicability of these primers were tested using 115 bacterial strains and fish tissues infected with S. iniae. Sensitivity, reproducibility and efficiency of qPCR assay were also determined. The developed qPCR assay showed 100% specificity with pure bacterial cultures or DNA extracted from S. iniae or tissues of fish infected with the bacterium. The method has high sensitivity with a detection limit of 1.12 × 101 amplicon copies per assay (equivalent to 2 × 10–9 ng/µl) using bacterial DNA and of 1.44 × 101 gene copies in tissues of fish infected with S. iniae. In conclusion, this qPCR protocol provides an accurate and sensitive alternative for the identification of S. iniae and its detection on fish tissues that can be implemented as a routine tool in microbiological laboratories.  相似文献   

4.
Rainbow trout from 23 families were evaluated for growth and resistance to the bacterial coldwater disease (BCWD) caused by Flavobacterium psychrophilum and infectious haematopoietic necrosis (IHN) caused by IHN virus. Average family weights were between 161 and 263 g with an average of 225 g at 213 days post‐fertilization with specific growth rates ranging from 2.37 to 2.88. Per cent survival of fish challenged with F. psychrophilum was between 18% and 100%, while for those challenged with IHNV, the range was between 12% and 93%. Significant positive correlations were found for end body weight and resistance to IHN (P < 0.05) and for early body weight and resistance to BCWD (P < 0.1). However, no significant correlations were detected between resistance to both pathogens or disease resistance and overall genetic diversity or diversity within the major histocompatibility locus.  相似文献   

5.
A previously developed monoclonal antibody (mAb) specific for the heavy chain of hybrid striped bass (HSB) Morone chrysops×M. saxatilis immunoglobulin was used in an assay to detect the humoral response to antigens of Streptococcus iniae. In order to validate this assay, an anti‐S. iniae antibody was produced in HSB by immunization with formalin‐killed cells of S. iniae mixed with Freund's complete adjuvant (FCA). After boosting with cells mixed with Freund's incomplete adjuvant (FIA), fish were challenged with live S. iniae by i.p. injection. This resulted in mortalities of 100%, 13% and 7% for non‐immunized, immunized and non‐challenged fish respectively. Live S. iniae were recovered only from moribund non‐immunized challenged fish, indicating that the immunization conferred protection against S. iniae. Sera were taken at 28 and 42 days post challenge and anti‐S. iniae antibody was measured by both agglutination and indirect ELISA. Titres of anti‐S. iniae antibody increased only in the immunized group as measured by agglutination and ELISA. Serum complement measured in the final bleeding was significantly higher in the immunized group. Western blotting using immune HSB serum indicated that the predominant antigen in this case was the high molecular weight polysaccharide of S. iniae.  相似文献   

6.
Vaccination strategies have traditionally been used as preventative or prophylactic measures against disease (prophylactic immunization) in uninfected fish. Alternatively, therapeutic or remedial measures, such as antibiotic administration, are commonly employed to treat disease in infected fish. Vaccination as a therapeutic measure (therapeutic immunization), however, has not been adequately explored in sub‐clinically infected fish. Therapeutic and prophylactic immunization with three Streptococcus iniae vaccines, formalin‐killed whole S. iniae cells (FKC vaccine), concentrated S. iniae extracellular products (greater than 2 kDa) (ECP vaccine) and a combination of killed cells and extracellular products (FKC+ECP vaccine), were tested in hybrid striped bass, Morone chrysops×Morone saxatilis, previously naturally infected with S. iniae. Fish (mean weight 10.0 g) were injected intraperitoneally (IP) or intramuscularly (IM) with one of each of the vaccines, tryptic soy broth (TSB‐control) or non‐injected (non‐injected control) to evaluate therapeutic effects (Trial 1). Survivors of the natural infection and ECP and FKC+ECP vaccine immunization and another lot of non‐injected control fish were immersion challenged with 1.47 × 106 CFU of S. iniae mL?1 at 44 days post‐immunization to evaluate vaccine efficacy (Trial 2). Hybrid striped bass (1.0 g) were also IM injected with S. iniae ECP vaccine at an aquaculture facility and immersion challenged with 1.47 × 106 CFU of S. iniae mL?1 12 weeks post‐immunization (Trial 3). The ECP and FKC+ECP vaccines, regardless of injection route, significantly (P<0.001) increased survival in asymptomatic, sub‐clinically infected fish thereby providing therapeutic merit. Hybrid bass immunized IP or IM had mean per cent survival values ranging from 78 to 96 at 44 days post‐immunization (Trial 1) and 69–97 post challenge (Trial 2). Survival of fish injected with TSB or immunized with FKC vaccine was significantly lowered and ranged from 12 to 13 by IP injection and 40 to 50 by IM injection and thus, the FKC vaccine had no therapeutic effect. The survival of hybrid striped bass IM immunized with S. iniae ECP vaccine in field Trial 3 was 91 and the RPS was 83. These results demonstrate that therapeutic immunization using S. iniae ECP and FKC+ECP vaccines can control a natural S. iniae infection. Furthermore, S. iniae ECP or FKC+ECP vaccines can also be used prophylacticly to protect hybrid striped bass against subsequent pathogen challenge.  相似文献   

7.
This study was undertaken to assess the impacts of dietary astaxanthin supplementation on growth performance, feed utilization, survival, and serum growth hormone (GH) availability of Asian seabass, Lates calcarifer, with special reference to dose–response relationships and variations during different feeding phases (short‐term, medium‐term and long‐term). Fish were fed the following diets in triplicate for 90 days: the control (CD), AX50 (50 mg astaxanthin/kg diet), AX100 (100 mg astaxanthin/kg diet) and AX150 (150 mg astaxanthin/kg diet). The findings revealed that fish exhibited significant linear increments (p < .05) in specific growth rate (SGR), weight gain, feed utilization efficiency and survival when fed various diets with escalating levels of astaxanthin. Supplementation with dietary astaxanthin significantly augmented (p < .05) GH levels in fish. Significant positive associations (p < .05) were observed between circulating serum GH levels and SGR of fish from all groups following three consecutive feeding phases, denoting a robust cause‐and‐effect relationship. Circulating GH concentrations were considered as a sensitive biomarker of growth performance in Asian seabass. This study illustrated that supplemental astaxanthin could be administered in culture protocols to improve the growth rate and commercial hatchery production of Asian seabass, and possibly other teleost species.  相似文献   

8.
A feeding trial was conducted to study the effect of mannan oligosaccharide (MOS) supplementation on growth, body composition, haematology, biochemical parameters and histology of Asian seabass (Lates calcarifer) fingerlings (8.13 ± 0.06 g average weight). Mannan oligosaccharide was supplemented at five different concentrations viz., 0%, 0.5%, 1%, 1.5% and 2% levels in the diet (40% protein and 9% lipid) of L. calcarifer. The results of the 60 days feeding trial showed that there was a significant (P < 0.05) increase in the final body weight, weight gain (WG), WG%, average daily gain, survival, specific growth rate, daily growth coefficient, hepatosomatic index and viscerosomatic index in the fish fed diet containing 1% MOS. Whole body composition of post‐fed animals showed non‐significant differences (P > 0.05) among the various treatment groups. The analysis of haematological parameters showed that there was no significant (P > 0.05) differences among different treatments but for the haemoglobin content which was significantly (P < 0.05) higher in the fish fed 1% MOS. Biochemical parameters revealed that glucose, urea, cholesterol and triglyceride content showed significant (P < 0.05) difference between control and MOS‐supplemented group. Histological observations of post‐fed animals revealed that MOS supplementation resulted in increased absorptive surface area of the intestine and increased glycogen deposition in liver. The result of this experiment infers that MOS supplementation has got a beneficial effect in the diet of seabass and supplementation at 1% level is optimal for improving the growth.  相似文献   

9.
Asian seabass (Lates calcarifer) has been recognized as an economically important aquaculture species which can be adapted to and cultivated in wide range of salinities. The number of freshwater intensive seabass farms in Thailand is increasing annually. Here, we first describe the susceptibility of Asian seabass, which were cultured in freshwater, to Streptococcus inae (SI) and their pathological changes. Three isolates of putative SI were identified using a combination of standard biochemical assays and species‐specific PCR prior subjected to in vivo challenge. Accumulated mortalities of the fish which received 107 CFU fish?1 of either SI1J, SI SGSA or SI2J were 90%, 90% and 100% at 7 days‐post infection (dpi), respectively, and mortalities increased sharply between 3 and 5 dpi. Clinical signs such as erratic swimming and opaque eyes were identified from a few infected fish, while most died rapidly without any abnormal signs. Histopathological manifestations were observed in the multiple organs (kidney, liver and brain). Haemorrhage, hyperhemia, cellular degeneration and inflammatory cells infiltration were commonly found within the internal organs. Notably, the formation of numerous encyst‐like lesion aggregated by eosinophilic cells, resembling macrophages, were typically found in the brain of the infected fish. Summarily, this study first revealed that freshwater reared Asian seabass is highly susceptible to SI infection and haemorrhagic septicaemia was a major pathological change that could be found in the infected fish.  相似文献   

10.
This is the first study to isolate, identify and characterize Streptococcus iniae as the causative disease agent in two tilapia (Oreochromis aureus) populations. The populations were geographically isolated, of distinct origins, and did not share water sources. Affected fish showed various external (e.g., exophthalmia and cachexia, among others) and internal (e.g., granulomatous septicaemia and interstitial nephritis, among others) signs. All internal organ samples produced pure cultures, two of which (one from each farm, termed S‐1 and S‐2) were subjected to biochemical, PCR and 16S rRNA sequencing (99.5% similarity) analyses, confirming S. iniae identification. The two isolates presented genetic homogeneity regardless of technique (i.e., RAPD, REP‐PCR and ERIC‐PCR analyses). Pathogenic potentials were assessed through intraperitoneal injection challenges in rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). Rainbow trout mortalities were respectively 40% and 70% at 104 and 106 CFU per fish with the S‐1 isolate, while 100% mortality rates were recorded in zebrafish at 102 and 104 CFU per fish with the S‐2 isolate. The obtained data clearly indicate a relationship between intensified aquaculture activities in Mexico and new disease appearances. Future studies should establish clinical significances for the tilapia industry.  相似文献   

11.
The Pacific oyster Crassostrea gigas is a representative bivalve mollusc that is widely cultured in the world and is the largest molluscan group cultured in China. In order to assess the feasibility of improving survival of C. gigas through genetic selection, the heritability and genetic correlations for growth and survival traits between different life stages were examined. Genetic parameters were estimated based on intraclass correlations of 49 full‐sib families (29 half‐sib families) in larvae (4 and 20 days after fertilization) and spat (140 days after fertilization) stages. The heritability for growth traits in larvae and spat was 0.30–0.86 and 0.53–0.59, respectively, and varied with ages. The heritability of survival was low in larvae (0.13 ± 0.05 and 0.17 ± 0.04, respectively for 4 and 20 days after fertilization) but medium (0.39 ± 0.07) in spat, suggesting that selection for increasing spat survival was feasible. The genetic correlation between growth traits within age was medium to high and positive (ranging from 0.47 to 0.96, respectively, between shell length (SL) and shell height (SH) at 20 days and between SL and SH at 140 days after fertilization), suggesting that selection to improve single growth trait will cause positive response in another growth traits in C. gigas. The genetic correlations between survival and growth traits at 140 days were low but positive (ranging from 0.23 to 0.27, respectively, between survival and SH and between survival and SL at 140 days after fertilization), suggesting that selection for survival may not have a negative response in growth. Overall, this study suggests that survival traits should be taken as improving target of next selection breeding programme in C. gigas.  相似文献   

12.
Francisella orientalis is a highly virulent, emerging bacterium that causes mass mortalities in tilapia. This pathogen also affects numerous other warm-water fish species, including three-line grunt, hybrid striped bass and various ornamental fish. This study sheds light on two new species of fish that are susceptible to F. orientalis. Asian seabass and largemouth bass showed variable levels of susceptibility in a bacterial challenge experiment. After intraperitoneally injected with a dose of 106 CFU/fish, a total of 64.28% and 21.42% mortalities were obtained in Asian seabass and largemouth bass, respectively. Meanwhile, Nile tilapia showed acute mortality of 100%. All fish showed typical lesions of francisellosis, including multifocal granulomas in the spleen and head kidney. Immunohistochemical analysis revealed strong positive signals inside the granulomas of all fish. The bacterial recovery in solid media from infected fish was highest in Nile tilapia (85.71%), followed by Asian seabass (35.71%) and largemouth bass (21.42%). PCR results tested 100% positive for Nile tilapia, and 78.57% and 21.42% for Asian seabass and largemouth bass, respectively. In conclusion, Asian seabass and largemouth bass are susceptible to this pathogen, which warrants new management strategies when employing predation polyculture systems of these species with tilapia.  相似文献   

13.
This study determined the growth performance and acquired resistance of Nile tilapia, Oreochromis niloticus (L.) that survived Streptococcus iniae infection. Tilapia were challenged with three doses of S. iniae (8.8 × 103, 8.8 × 104 and 8.8 × 105 CFU fish?1 for low, medium and high challenges respectively). Groups of non‐injected and tryptic soy broth‐injected fish were maintained as controls. Significantly (P<0.05) higher mortality (45.0%) occurred in the high challenge treatment than in the low challenge treatment group (29.6%). The medium challenge group had mortality (36.3%) that did not differ significantly from the high or low treatment. Few fish died in the non‐injected and broth‐injected treatments (3.4% and 0.8% respectively). The tilapia that survived S. iniae infection used to assess growth performance were selected from survivors without gross clinical signs of disease. These fish were randomly stocked at a rate of 30 fish into each 57 L aquarium in triplicate and fed to apparent satiation for 8 weeks. No significant differences were detected in weight gain, feed intake, feed efficiency ratio or survival between S. iniae‐survived tilapia and the control treatments following the 8‐week growth performance trial. Following the 8‐week feeding study, tilapia were challenged with 1 × 106 CFU fish?1 of S. iniae to assess acquired immunity. Mean cumulative mortality was significantly higher (P<0.05) in the control treatments (41.7% for the non‐injected and 43.3% for the broth‐injected fish) than in the low, medium and high challenge treatments (7.4%, 3.3% and 8.3% respectively). Serum protein was significantly (P<0.05) elevated in the S. iniae‐survived tilapia that were subsequently challenged when compared with controls challenged for the first time. Agglutinating antibody titre was significantly higher in the fish in the medium and high challenge treatments, compared with the control fish challenged for the first time. The results suggest tilapia that survive S. iniae challenge without showing overt disease signs performed as well as non‐infected tilapia. Further, the S. iniae‐survived tilapia challenged following the 8‐week growth performance trial gained acquired resistance to homologous S. iniae challenge.  相似文献   

14.
This study was conducted to characterize the causative agent of streptococcosis in tilapia (Oreochromis niloticus) and control of Streptococcus infection by means of garlic (Allium sativum) supplementation. The morphological, biochemical and polymerase chain reaction amplification confirmed 11 isolates belong to Streptococcus iniae from the infected fish eyes and tissue samples. Random screening of 12 well‐known medicinal plant parts against S. iniae revealed the garlic extract as the most effective herbal recovery against Streptococcus infection. In vivo challenge test with dietary supplementation of garlic powder significantly improved survival rates of fish against S. iniae infections, and modulate the microbial community and cytokine gene expression profiling in the intestine of the experimental tilapia. Among the two garlic supplemented treatments, 1.0 g garlic supplemented diet significantly increased (p < 0.05) the survival rates of tilapia and the gut bacterial operational transitional units abundance for Proteobacteria and Tenericutes, the phyla associated with healthy intestinal flora. The bacterial diversity index also found high with garlic supplemented diets. Significant upregulations of IL‐10 and IL‐17F gene expression in the intestinal tissue were observed with 1.0 g garlic supplemented diet where IL‐8 and IL‐1β expression levels were relatively static. The dietary supplementation of garlic, therefore, could be effective in the prevention of S. iniae infection in fish.  相似文献   

15.
The antimicrobial activity and mode of action of chitosan were evaluated against Streptococcus iniae, a pathogenic Gram‐positive bacterium of fish worldwide. Cell proliferation kinetics were examined following exposure to varying concentrations of chitosan. The action of chitosan on S. iniae was also investigated by measuring agglutination activity, conductivity, and extracellular and intracellular bacterial adenosine triphosphate (ATP) levels. Chitosan exhibited antibacterial activity against S. iniae at concentrations of 0.1% and above and was lethal at a concentration of 0.4% and higher. The mechanism of antibacterial activity of chitosan at the inhibitory level of bacterial growth appears to hinge upon the interaction between chitosan and the oppositely charged bacterial surface. This interplay causes agglutination, which was readily observed grossly and microscopically. After interacting with the cell surface via adsorption, an efflux of intracellular ATP was documented, which suggests that chitosan disrupts the bacterial cell causing leakage of cytosolic contents and ultimately cell death. Results suggest chitosan may be worth evaluating as a natural alternative to antibiotic against S. iniae infection of fish.  相似文献   

16.
Heritabilities and genetic and phenotypic correlations were estimated for body weight (BW), body length (BL), carapace length (CL), carapace width (CW) and carapace height (CH) of the Penaeus monodon from measurements on progeny at 6 months of age resulting from a nested mating design. Estimation was made on data from 41 full‐sib and 10 half‐sib families. The estimated heritabilities for these traits were 0.13–0.24, and the significantly phenotypic and genetic correlations were 0.81–0.94 and 0.88–1.00, respectively. The present results indicate that genetic improvement through selective breeding conducted on P. monodon might be feasible and would be an efficient method to obtain a positive response to selection.  相似文献   

17.
In this study, four essential oils—cinnamon oil, leech lime oil, lemongrass oil, and turmeric oil—were examined for their antimicrobial activities against Streptococcus iniae, a bacterium that is pathogenic in fish, in which it causes streptococcosis. Cinnamon oil was the most potent antimicrobial agent among these oils, with a minimal inhibitory concentration (MIC) of 40 μg/ml. By using gas chromatography–mass spectrometry (GC–MS), it was found that the major components of cinnamon oil were cinnamaldehyde (90.24), limonene (2.42%), cinnamyl acetate (2.03%), linalool (1.16%), and α-terpineol (0.87%). Of these compounds, only cinnamaldehyde exhibited antimicrobial activity against S. iniae, with an MIC of 20 μg/ml. In an in vivo trial, no mortality was apparent in fish fed on fish diets supplemented with 0.4% (w/w) of cinnamon oil and with 0.1% (w/w) of oxytetracycline 5 days prior to infection with S. iniae. These results indicate that cinnamon oil had a protective effect on experimental S. iniae infection in tilapia, and thus has the potential to replace the antibiotics used to control this disease.  相似文献   

18.
Cannibalism has been described in 390 teleost fish species belonging to 104 families. In rearing conditions, intra‐cohort cannibalism is one of the major bottlenecks during the early life stages of pikeperch (Sander lucioperca). This study aims to describe precisely the occurrence and the onset of cannibalism of this species during the first two post‐hatching months. In a first step, the cannibalistic behaviour was analysed through observations in three experiments. In each experiment, the number of cannibalistic cases was similar. From 14 to 48 days post‐hatching (dph), more than 60 % of prey ingestions were realized by tail‐first, but after 48 dph, the number of prey ingestions by head‐first increased. In a second step, the behavioural sequence exhibited to attack and to capture a conspecific was analysed, taking into account the size ratio between the cannibal and its prey. The behavioural sequence to capture was always the same, similar to a predatory sequence and it seems that the cannibal chose its prey about 10 s before attacking. Moreover, prey choice by a cannibal was based more on the size ratio between the cannibal and its prey than the previous activity of the potential prey. These new results could lead to solutions to decrease cannibalism in pikeperch rearing.  相似文献   

19.
A long-term environmental study was conducted to evaluate the significance of water temperature, pH and salinity on the prevalence of Streptococcus iniae mortalities in barramundi sea-cage facilities. It was determined that there is a strong association between temperature and increased mortality, specifically between 25 and 28 °C. Temperatures outside of that range result in decreased mortalities attributed to S. iniae. There was no statistical significance between pH or salinity and S. iniae induced mortality (p > 0.05), although acidic conditions (< pH 6) occurring due to anthropogenic disturbances did result in acute mortalities and S. iniae was recovered from approximately 70% these fish. Laboratory challenge studies confirmed the temperature dependence of S. iniae infections as well as the increased susceptibility of barramundi to S. iniae during acid water conditions.  相似文献   

20.
Skipjack tuna viscera hydrolysate (TVH) was produced by enzymatic hydrolysis using alcalase. A study of the effect of time and enzyme concentration was performed and formed the basis for preparing TVH with the desired degree of hydrolysis (DH). An enzyme to substrate ratio of 5 g kg−1 (TVH A1) and 16.4 g kg−1 (TVH A2) at pH 8.0 with a temperature of 50 °C, and hydrolysis time of 60 min were found to be optimal conditions to obtain hydrolysis of 42.9% and 62.9%, respectively. Both TVH A1 and TVH A2 were analysed for chemical and amino acid composition and peptide size. The effectiveness of TVH at different DH as feeding attractants was estimated in a feeding trial at different inclusion levels. Ten experimental diets , including a fish meal‐based reference diet, were fed to Asian seabass for 28 days. Fish fed the diet supplemented with 30 g kg−1 of TVH A2, and the reference diet showed the highest feed intake (P < 0.05). The growth performance of fish fed diets supplemented with TVH was slightly higher than those of fish fed the control diet, but the reference group showed a significantly higher growth rate (P < 0.05). These findings suggest that the supplementation of feed with TVH that has been processed under suitable conditions can improve feed palatability and growth performance when utilizing alternative protein sources in Asian seabass diets.  相似文献   

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