Pacific white shrimp has become a major aquaculture and fishery species worldwide.Although a large scale EST resource has been publicly available since 2008,the data have not yet been widely used for SNP discovery or ...Pacific white shrimp has become a major aquaculture and fishery species worldwide.Although a large scale EST resource has been publicly available since 2008,the data have not yet been widely used for SNP discovery or transcriptome-wide assessment of selective pressure.In this study,a set of 155 411 expressed sequence tags(ESTs) from the NCBI database were computationally analyzed and 17 225 single nucleotide polymorphisms(SNPs) were predicted,including 9 546 transitions,5 124 transversions and 2 481 indels.Among the 7 298 SNP substitutions located in functionally annotated contigs,58.4%(4 262) are non-synonymous SNPs capable of introducing amino acid mutations.Two hundred and fifty nonsynonymous SNPs in genes associated with economic traits have been identified as candidates for markers in selective breeding.Diversity estimates among the synonymous nucleotides were on average 3.49 times greater than those in non-synonymous,suggesting negative selection.Distribution of non-synonymous to synonymous substitutions(Ka/Ks) ratio ranges from 0 to 4.01,(average 0.42,median 0.26),suggesting that the majority of the affected genes are under purifying selection.Enrichment analysis identified multiple gene ontology categories under positive or negative selection.Categories involved in innate immune response and male gamete generation are rich in positively selected genes,which is similar to reports in Drosophila and primates.This work is the first transcriptome-wide assessment of selective pressure in a Penaeid shrimp species.The functionally annotated SNPs provide a valuable resource of potential molecular markers for selective breeding.展开更多
The aim of the present study is to detect the potential of the base population from diallel crosses of eight introduced strains of the Pacific white shrimp（Litopenaeus vannamei） for improving the yield. Heterosis an...The aim of the present study is to detect the potential of the base population from diallel crosses of eight introduced strains of the Pacific white shrimp（Litopenaeus vannamei） for improving the yield. Heterosis and heritability were estimated for pond survival at commercial farm conditions for the base population that included 207 full-sib families from a nested mating design by artificial insemination. Among all the hybrids,the heterosis ranged from –11.37%（UA1×UA2） to 20.53%（UA3×SIN） with an average of 0.953%. The results showed that more than half of the hybrids（51.85%） have negative heterosis for survival rate, but most of the hybrids with positive heterosis have high estimates. The high proportion of negative heterosis for survival rate reminders us that the survival trait also should be considered in the crossbreeding program to avoid yield decrease. However, high positive heterosis manifested in most of the hybrids for survival indicates the usefulness of these hybrids for improving the survival to obtain higher yield by crossbreeding in this breeding program. The heritability estimate for pond survival was 0.092±0.043 when genetic groups were included in the pedigree, and it was significantly different from zero（P〈0.05）. The results from this study also indicated that significant improvement for survival is possible through selection in L. vannamei.展开更多
Fortilin is a multifunctional protein implicated in many important cellular processes. Since injection of Pm-fortilin reduces shrimp mortality caused by white spot syndrome virus (WSSV), there is potential application...Fortilin is a multifunctional protein implicated in many important cellular processes. Since injection of Pm-fortilin reduces shrimp mortality caused by white spot syndrome virus (WSSV), there is potential application of fortilin in shrimp culture. In the present study, in order to improve trans-membrane transportation efficiency, the protein transduction domain of the transactivator of transcription (TAT) peptide was fused to fortilin. The Pichia pastoris yeast expression system, which is widely accepted in animal feeds, was used for production of recombinant fusion protein. Green fluorescence protein (GFP) was selected as a reporter because of its intrinsic visible fluorescence. The fortilin, TAT and GFP fusion protein were constructed. Their trans-membrane transportation efficiency and effects on immune response of shrimp were analyzed in vitro. Results showed that TAT peptide improved in vitro uptake of fortilin into the hemocytes and midgut of Litopenaeus vannamei. The phenoloxidase (PO) activity of hemocytes incubated with GFP-Fortilin or GFP-Fortilin-TAT was significantly increased compared with that in the control without expressed fortilin. The PO activity of hemocytes incubated with 200 μg mL-1 GFP-Fortilin-TAT was significantly higher than that in the group with the same concentration of GFP-Fortilin. Hemocytes incubated with GFP-Fortilin-TAT at all concentrations showed significantly higher nitric oxide synthase (NOS) activity than those in the control or in the GFP-Fortilin treatment. The present in vitro study indicated that TAT fusion protein improved the immune effect of fortilin.展开更多
White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacter...White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect L itopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus(WSSV+Vp), or we fi rst infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection(Vp+WSSV). The effect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of V ibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction(PCR) method. L v My D88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the different mortality rates. Our results show that(1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection;(2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed signifi cant increases in the numbers of Vibrio( P <0.05);(3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10 5 to 10 7 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.展开更多
The optimal quantity of dietary lysine (Lys) and arginine (Arg), and the optimal ratio of dietary Lys to Arg for Pacific white shrimp Litopenaeus vannamei were investigated. Coated Lys and Arg were added to a basal di...The optimal quantity of dietary lysine (Lys) and arginine (Arg), and the optimal ratio of dietary Lys to Arg for Pacific white shrimp Litopenaeus vannamei were investigated. Coated Lys and Arg were added to a basal diet (37.99% crude protein and 7.28% crude lipid) to provide graded levels of Lys and Arg. The experimental diets contained three Lys levels (2.51%, 2.11%, and 1.70% of total diet), and three Arg levels (1.41%, 1.80%, and 2.21% of total diet) and all combinations of these levels were tested. Pacific white shrimp, with a mean weight of 3.62±0.1 g, were randomly distributed in 36 fiberglass tanks with 30 shrimp per tank and reared on the experimental diets for 50 days. After the feeding trial, the growth performance, survival, feed conversion rate (FCR), body composition and protease and lipase activities in the hepatopancreases of the experimental shrimps were determined. The results show that weight gain (WG), specific growth rate (SGR), FCR, body protein, body Lys and Arg content were significantly affected by dietary Lys and Arg (P<0.05) and improved when dietary Lys and Arg levels were 2.11% 2.51% and 1.80% 2.21%, respectively. Protease and lipase activities in the hepatopancreases of the shrimps appeared higher when dietary Lys and Arg quantities were 2.11% 2.51% and 1.80% 2.21%, although the difference was not statistically significant (P>0.05). Therefore, according to our results, the optimal Lys and Arg quantities in the diet of Pacific white shrimp, L. vannamei, were considered to be 2.11%-2.51% and 1.80%- 2.21%, respectively, and the optimal ratio to be 1:0.88-1:1.05.展开更多
In this study, specific growth rate(SGR), ingestion rate(IR), food conversion ratio(FCR), apparent digestion ratio(ADR) and ammonium-nitrogen excretion were determined for sea cucumber(Stichopus monotuberculatus) rear...In this study, specific growth rate(SGR), ingestion rate(IR), food conversion ratio(FCR), apparent digestion ratio(ADR) and ammonium-nitrogen excretion were determined for sea cucumber(Stichopus monotuberculatus) reared in plastic containers(70 L; 4 containers each diet treatment). Sea cucumbers were fed with five diets containing different amounts of farming waste from shrimp(Litopenaeus vannamei)(100%, 75%, 50%, 25% and 0) and a formulated compound(20% sea mud and 80% powdered algae). Sea cucumbers grew faster when they were fed with diet D(25% shrimp waste and 75% formulated compound) than those fed with other diets. Although IR value of sea cucumber fed with diet A(shrimp waste) was higher than those fed with other diets, both the lowest SGR and the highest FCR occurred in this diet group. The highest and the lowest ADR occurred in diet E(formulated compound) and diet A group, respectively, and the same to ammonium-nitrogen excretion. The contents of crude protein, crude lipid and total organic matter(TOM) in feces decreased in comparison with corresponding diets. In the feces from different diet treatments, the contents of crude protein and TOM increased gradually as the contents of crude protein and TOM in diets increased, while crude lipid content decreased gradually as the crude lipid content in diets increased.展开更多
In the present study, the applicability of the wet waste collected from shrimp(Litopenaeus vannamei) to the culture of sea cucumber(Stichopus monotuberculatus) was determined. The effects of dietary wet shrimp waste o...In the present study, the applicability of the wet waste collected from shrimp(Litopenaeus vannamei) to the culture of sea cucumber(Stichopus monotuberculatus) was determined. The effects of dietary wet shrimp waste on the survival, specific growth rate(SGR), fecal production rate(FPR), ammonia- and nitrite-nitrogen productions of sea cucumber were studied. The total organic matter(TOM) level in the feces of sea cucumber was compared with that in corresponding feeds. Diet C(50% wet shrimp waste and 50% sea mud mash) made sea cucumber grow faster than other diets. Sea cucumber fed with either diet D(25% wet shrimp waste and 75% sea mud mash) or sole sea mud exhibited negative growth. The average lowest total FPR of sea cucumber occurred in diet A(wet shrimp waste), and there was no significant difference in total FPR between diet C and diet E(sea mud mash)(P > 0.05). The average ammonia-nitrogen production of sea cucumber in different diet treatments decreased gradually with the decrease of crude protein content in different diets. The average highest nitrite-nitrogen production occurred in diet E treatment, and there was no significant difference in nitrite-nitrogen production among diet A, diet B(75% wet shrimp waste and 25% sea mud mash) and diet C treatments(P > 0.05). In each diet treatment, the total organic matter(TOM) level in feces decreased to different extent compared with that in corresponding feeds.展开更多
The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus （WSSV） in shrimp. In this...The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus （WSSV） in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei （LvPT） was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigenTM design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at -27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA （4 pg per shrimp） was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection （hpi）. Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.展开更多
Microfungi have been explored further as supplement constituents in shrimp culture due to their rich protein,lipid,essential amino acids,fatty acids,and various bioactive compounds.The researchers have evaluated the e...Microfungi have been explored further as supplement constituents in shrimp culture due to their rich protein,lipid,essential amino acids,fatty acids,and various bioactive compounds.The researchers have evaluated the effects of two microfungi powders(Mucor circinelloides and Trichoderma harzianum)as feed supplements on growth performance,bacterial communities,and the biomass nutritional profile of white shrimp(Litopenaeus vannamei).The results showed that commercial/artificial feed(Feng-Li)with supplementation microfungi powder had a different effect on white shrimp growth performance than control after 62 days of the grow-out period.Supplementation with 1.5%M.circinelloides powder(M-2)could improve the growth performance of white shrimp,especially the survival rate(81.33%),final body weight(5.21 g),and shrimp biomass(211.35 g).In contrast,supplementation with 1.5%T.harzianum powder(T-2)did not improve the growth performance of white shrimp compared to control(p>0.05).Furthermore,it also affected the profile of the bacterial community in culture water as well as in shrimp intestines.Supplementation with both microfungi could reduce the total number of bacteria in culture water at the end of the experiment,where the lowest number was obtained in treatment T-2(3.9×10^(5)CFU/mL),followed by M-2(9.9×10^(5)CFU/mL)and control(4.4×10^(6)CFU/mL).In the culture water,20 isolates of culturable bacteria were found during the trial,with the dominant bacteria in M-2:Bacillus cereus,Bacillus velezensis,Gordonia polyisoprenivorans,T-2:Acinetobacter sp.,Gordonia polyisoprenivorans,Micrococcus luteus.Based on average well colour development(AWCD),the highest bacterial activity was in the M-2(0.297-0.707),followed by T-2(0.226-0.717)and control(0.094-0.694).The highest substrate group utilised in M-2 and T-2 were polymers(0.896 and 1.348)and control:amines(0.100).The highest Shannon diversity index was obtained in the M-2(2.508),followed by control(2.286)and the T-2(2.157).As many as 11 isolates of culturable bacteria were found in the shrimp intestine,with dominant bacteria in M-2:Bacillus velezensis,Micrococcus luteus,Bacillus cereus,T-2:Bacillus velezensis,Micrococcus sp.2,Demequina activiva.The highest bacterial activity was in M-2(2.259),followed by control(2.020)and T-2(0.050).The highest substrate group utilisation was in M-2:polymers(2.564),T-2:carbohydrates(0.094),and control:polymers(2.185).The highest Shannon diversity index was in the M-2(3.234),followed by control(3.230)and the T-2(0.751).In addition,1.5%M.circinelloides and T.harzianum supplementation increased white shrimp muscle protein content up to 79.21%and 80.40%,respectively,compared to control(76.73%).In conclusion,using Mucor circinelloides powder as a feed supplement can improve the growth performance of white shrimp,increase the functional/probiotic bacteria:B.cereus and B.velezensis in cultur and shrimp’s intestine,and increase the protein content in shrimp muscle tissue.However,more in-depth research is needed,especially on anti-nutritional compounds from fungi that affect shrimp growth.展开更多
The cultivation of the marine species of shrimp Litopenaeus vannamei has emerged as one of the segments that best develop in the Brazilian aquaculture, representing the agribusiness that grew the most in recent years....The cultivation of the marine species of shrimp Litopenaeus vannamei has emerged as one of the segments that best develop in the Brazilian aquaculture, representing the agribusiness that grew the most in recent years. Regarding the economic importance of the farming of this species in Brazil, further studies on genetic improvement were conducted. A F2 segregating population consisting of 192 samples for each G1 and G2 families, from crossing inbred lines was used for the studies performed in the present work. The genetic linkage analysis was based on polymorphic markers derived from nine AFLP (Amplified Fragment Length Polymorphism) primers. Fourteen genetic linkage groups including 103 segregating polymorphic markers were constructed covering350 cMfor G1 and four genetic linkage groups including 59 markers were constructed covering300 cMfor G2. Simple marker analyses were performed among individuals evaluated phenotypically, finding markers linked to genes that may be potentially important and useful to assess characteristics of economic importance for traits related to weight and the disease IMN (idiopathic muscle necrosis). The statistical model including markers explained major proportion of phenotypic characteristic weight in relation to disease incidence IMN.展开更多
The phosphatidylinositol 3-kinase(PI3K)-AKT pathway is involved in various cellular functions, including anti-apoptosis, protein synthesis, glucose metabolism and cell cycling. However, the role of the PI3K-AKT pathwa...The phosphatidylinositol 3-kinase(PI3K)-AKT pathway is involved in various cellular functions, including anti-apoptosis, protein synthesis, glucose metabolism and cell cycling. However, the role of the PI3K-AKT pathway in crustaceans remains unclear. In the present study, we cloned and characterized the AKT gene lvakt from Litopenaeus vannamei. The 511-residue LVAKT was highly conserved; contained a PH domain, a catalytic domain and a hydrophobic domain; and was highly expressed in the heart and gills of L. vannamei. We found, using Real-Time Quantitative PCR(Q-PCR) analysis, that lvakt was upregulated during early white spot syndrome virus(WSSV) infection. Moreover, the PI3K-specific inhibitor, LY294002, reduced viral gene transcription, implying that the PI3K-AKT pathway might be hijacked by WSSV. Our results therefore suggest that LVAKT may play an important role in the shrimp immune response against WSSV.展开更多
Anti-lipopolysaccharide factors （ALFs） are basic components of the crustacean immune system that defend against a range of pathogens. The cDNA sequence of a new ALF, designated nLvALF2, with an open reading flame en...Anti-lipopolysaccharide factors （ALFs） are basic components of the crustacean immune system that defend against a range of pathogens. The cDNA sequence of a new ALF, designated nLvALF2, with an open reading flame encoding 132 amino acids was cloned. Its deduced amino acid sequence contained the conserved functional domain of ALFs, the LPS binding domain （LBD）. its genotnic sequence consisted of three exons and four introns, nLvALF2 was mainly expressed in the Oka organ and gills of shrimps. The transcriptional level of nLw4LF2 increased significantly after white spot syndrome virus （WSSV） infection, suggesting its important roles in protecting shrimps from WSSV. Single nucleotide polymorphisms （SNPs） were found in the genomic sequence of nLvALF2, of which 38 were analyzed for associations with the susceptibility/resistance of shrimps to WSSV. The loci g.2422 A〉G, g.2466 T〉C, and g.2529 G〉A were significantly associated with the resistance to WSSV （P〈0.05）. These SNP loci could be developed as markers for selection of WSSV-resistant varieties ofLitopenaeus vannamei.展开更多
基金Supported by the Key Program of National Natural Science Foundation of China (No.30730071)the National High Technology R&D Program of China(863 Program)(No.2012AA10A404)the Agricultural Science and Technology Achievements Transformation Funds Project(No.2010GB24910700)
文摘Pacific white shrimp has become a major aquaculture and fishery species worldwide.Although a large scale EST resource has been publicly available since 2008,the data have not yet been widely used for SNP discovery or transcriptome-wide assessment of selective pressure.In this study,a set of 155 411 expressed sequence tags(ESTs) from the NCBI database were computationally analyzed and 17 225 single nucleotide polymorphisms(SNPs) were predicted,including 9 546 transitions,5 124 transversions and 2 481 indels.Among the 7 298 SNP substitutions located in functionally annotated contigs,58.4%(4 262) are non-synonymous SNPs capable of introducing amino acid mutations.Two hundred and fifty nonsynonymous SNPs in genes associated with economic traits have been identified as candidates for markers in selective breeding.Diversity estimates among the synonymous nucleotides were on average 3.49 times greater than those in non-synonymous,suggesting negative selection.Distribution of non-synonymous to synonymous substitutions(Ka/Ks) ratio ranges from 0 to 4.01,(average 0.42,median 0.26),suggesting that the majority of the affected genes are under purifying selection.Enrichment analysis identified multiple gene ontology categories under positive or negative selection.Categories involved in innate immune response and male gamete generation are rich in positively selected genes,which is similar to reports in Drosophila and primates.This work is the first transcriptome-wide assessment of selective pressure in a Penaeid shrimp species.The functionally annotated SNPs provide a valuable resource of potential molecular markers for selective breeding.
基金The National Natural Science Foundation of China under contract No.31302179the Taishan Scholar Program for Seed Industry+4 种基金the Introduction of International Advanced Agricultural Science and Technology Plan under contract No.2016-X39the Special Fund for Basic Scientific Research Business of Central Public Research Institutes under contract No.2015B04XK01China Postdoctoral Science Foundation under contract No.2015M572095Qingdao Scientific Research Foundation under contract No.14-2-4-52-jch5Jiangsu Province Science and Technology Support Project under contract No.BE2014414
文摘The aim of the present study is to detect the potential of the base population from diallel crosses of eight introduced strains of the Pacific white shrimp（Litopenaeus vannamei） for improving the yield. Heterosis and heritability were estimated for pond survival at commercial farm conditions for the base population that included 207 full-sib families from a nested mating design by artificial insemination. Among all the hybrids,the heterosis ranged from –11.37%（UA1×UA2） to 20.53%（UA3×SIN） with an average of 0.953%. The results showed that more than half of the hybrids（51.85%） have negative heterosis for survival rate, but most of the hybrids with positive heterosis have high estimates. The high proportion of negative heterosis for survival rate reminders us that the survival trait also should be considered in the crossbreeding program to avoid yield decrease. However, high positive heterosis manifested in most of the hybrids for survival indicates the usefulness of these hybrids for improving the survival to obtain higher yield by crossbreeding in this breeding program. The heritability estimate for pond survival was 0.092±0.043 when genetic groups were included in the pedigree, and it was significantly different from zero（P〈0.05）. The results from this study also indicated that significant improvement for survival is possible through selection in L. vannamei.
基金supported by the Special Fund for Agro-scientific Research in the Public Interest (No. 201103034)
文摘Fortilin is a multifunctional protein implicated in many important cellular processes. Since injection of Pm-fortilin reduces shrimp mortality caused by white spot syndrome virus (WSSV), there is potential application of fortilin in shrimp culture. In the present study, in order to improve trans-membrane transportation efficiency, the protein transduction domain of the transactivator of transcription (TAT) peptide was fused to fortilin. The Pichia pastoris yeast expression system, which is widely accepted in animal feeds, was used for production of recombinant fusion protein. Green fluorescence protein (GFP) was selected as a reporter because of its intrinsic visible fluorescence. The fortilin, TAT and GFP fusion protein were constructed. Their trans-membrane transportation efficiency and effects on immune response of shrimp were analyzed in vitro. Results showed that TAT peptide improved in vitro uptake of fortilin into the hemocytes and midgut of Litopenaeus vannamei. The phenoloxidase (PO) activity of hemocytes incubated with GFP-Fortilin or GFP-Fortilin-TAT was significantly increased compared with that in the control without expressed fortilin. The PO activity of hemocytes incubated with 200 μg mL-1 GFP-Fortilin-TAT was significantly higher than that in the group with the same concentration of GFP-Fortilin. Hemocytes incubated with GFP-Fortilin-TAT at all concentrations showed significantly higher nitric oxide synthase (NOS) activity than those in the control or in the GFP-Fortilin treatment. The present in vitro study indicated that TAT fusion protein improved the immune effect of fortilin.
基金Supported by the National Basic Research Program of China(973 Program)(No.2012CB114405)the Special Foundation Under the Construction Program for the“Taishan Scholarship”of Shandong Province of Chinathe Program for Chinese Outstanding Talents in Agricultural Scientific Research
文摘White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect L itopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus(WSSV+Vp), or we fi rst infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection(Vp+WSSV). The effect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of V ibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction(PCR) method. L v My D88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the different mortality rates. Our results show that(1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection;(2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed signifi cant increases in the numbers of Vibrio( P <0.05);(3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10 5 to 10 7 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.
基金Supported by the Research Foundation for Advanced Talents, Qingdao Agricultural University (No.631109)
文摘The optimal quantity of dietary lysine (Lys) and arginine (Arg), and the optimal ratio of dietary Lys to Arg for Pacific white shrimp Litopenaeus vannamei were investigated. Coated Lys and Arg were added to a basal diet (37.99% crude protein and 7.28% crude lipid) to provide graded levels of Lys and Arg. The experimental diets contained three Lys levels (2.51%, 2.11%, and 1.70% of total diet), and three Arg levels (1.41%, 1.80%, and 2.21% of total diet) and all combinations of these levels were tested. Pacific white shrimp, with a mean weight of 3.62±0.1 g, were randomly distributed in 36 fiberglass tanks with 30 shrimp per tank and reared on the experimental diets for 50 days. After the feeding trial, the growth performance, survival, feed conversion rate (FCR), body composition and protease and lipase activities in the hepatopancreases of the experimental shrimps were determined. The results show that weight gain (WG), specific growth rate (SGR), FCR, body protein, body Lys and Arg content were significantly affected by dietary Lys and Arg (P<0.05) and improved when dietary Lys and Arg levels were 2.11% 2.51% and 1.80% 2.21%, respectively. Protease and lipase activities in the hepatopancreases of the shrimps appeared higher when dietary Lys and Arg quantities were 2.11% 2.51% and 1.80% 2.21%, although the difference was not statistically significant (P>0.05). Therefore, according to our results, the optimal Lys and Arg quantities in the diet of Pacific white shrimp, L. vannamei, were considered to be 2.11%-2.51% and 1.80%- 2.21%, respectively, and the optimal ratio to be 1:0.88-1:1.05.
基金supported by the Key Project of National Science & Technology Pillar Program in 12th Five-year Plan (2011BAD13B02, 2012BAD18B03)the Science & Technology Promoting Project for Oceanic & Fishery in Guangdong Province (A201100D01, A201101 D02)+3 种基金the Knowledge Innovation Key Project of Chinese Academy of Sciences (KZCX2-EW-Q212)the comprehensive strategic cooperation project of Guangdong Province and Chinese Academy of Sciences (2012B09 1100269)the Cooperation Program of Guangdong Province and Chinese Academy of Sciences (2012B0911 00272)the Foundation for Distinguished Young Talents in Higher Education of Guangdong, China (2014KQNCX183)
文摘In this study, specific growth rate(SGR), ingestion rate(IR), food conversion ratio(FCR), apparent digestion ratio(ADR) and ammonium-nitrogen excretion were determined for sea cucumber(Stichopus monotuberculatus) reared in plastic containers(70 L; 4 containers each diet treatment). Sea cucumbers were fed with five diets containing different amounts of farming waste from shrimp(Litopenaeus vannamei)(100%, 75%, 50%, 25% and 0) and a formulated compound(20% sea mud and 80% powdered algae). Sea cucumbers grew faster when they were fed with diet D(25% shrimp waste and 75% formulated compound) than those fed with other diets. Although IR value of sea cucumber fed with diet A(shrimp waste) was higher than those fed with other diets, both the lowest SGR and the highest FCR occurred in this diet group. The highest and the lowest ADR occurred in diet E(formulated compound) and diet A group, respectively, and the same to ammonium-nitrogen excretion. The contents of crude protein, crude lipid and total organic matter(TOM) in feces decreased in comparison with corresponding diets. In the feces from different diet treatments, the contents of crude protein and TOM increased gradually as the contents of crude protein and TOM in diets increased, while crude lipid content decreased gradually as the crude lipid content in diets increased.
基金supported by the Key Projects in the National Science & Technology Pillar Program during the 12th Five-year Plan Period (2011BAD13B02, 2012BAD 18B03)the Science & Technology Promoting Projects for Oceanic & Fishery in Guangdong Province (A2011 00D01, A201101D02)Cooperation Program of Guangdong Province & Chinese Academy of Sciences (2012B091100272)
文摘In the present study, the applicability of the wet waste collected from shrimp(Litopenaeus vannamei) to the culture of sea cucumber(Stichopus monotuberculatus) was determined. The effects of dietary wet shrimp waste on the survival, specific growth rate(SGR), fecal production rate(FPR), ammonia- and nitrite-nitrogen productions of sea cucumber were studied. The total organic matter(TOM) level in the feces of sea cucumber was compared with that in corresponding feeds. Diet C(50% wet shrimp waste and 50% sea mud mash) made sea cucumber grow faster than other diets. Sea cucumber fed with either diet D(25% wet shrimp waste and 75% sea mud mash) or sole sea mud exhibited negative growth. The average lowest total FPR of sea cucumber occurred in diet A(wet shrimp waste), and there was no significant difference in total FPR between diet C and diet E(sea mud mash)(P > 0.05). The average ammonia-nitrogen production of sea cucumber in different diet treatments decreased gradually with the decrease of crude protein content in different diets. The average highest nitrite-nitrogen production occurred in diet E treatment, and there was no significant difference in nitrite-nitrogen production among diet A, diet B(75% wet shrimp waste and 25% sea mud mash) and diet C treatments(P > 0.05). In each diet treatment, the total organic matter(TOM) level in feces decreased to different extent compared with that in corresponding feeds.
基金Supported by the National Basic Research Program of China(973 Program)(No.2012CB114403)the China Agriculture Research System-47(CARS-47)
文摘The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus （WSSV） in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei （LvPT） was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigenTM design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at -27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA （4 pg per shrimp） was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection （hpi）. Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.
文摘Microfungi have been explored further as supplement constituents in shrimp culture due to their rich protein,lipid,essential amino acids,fatty acids,and various bioactive compounds.The researchers have evaluated the effects of two microfungi powders(Mucor circinelloides and Trichoderma harzianum)as feed supplements on growth performance,bacterial communities,and the biomass nutritional profile of white shrimp(Litopenaeus vannamei).The results showed that commercial/artificial feed(Feng-Li)with supplementation microfungi powder had a different effect on white shrimp growth performance than control after 62 days of the grow-out period.Supplementation with 1.5%M.circinelloides powder(M-2)could improve the growth performance of white shrimp,especially the survival rate(81.33%),final body weight(5.21 g),and shrimp biomass(211.35 g).In contrast,supplementation with 1.5%T.harzianum powder(T-2)did not improve the growth performance of white shrimp compared to control(p>0.05).Furthermore,it also affected the profile of the bacterial community in culture water as well as in shrimp intestines.Supplementation with both microfungi could reduce the total number of bacteria in culture water at the end of the experiment,where the lowest number was obtained in treatment T-2(3.9×10^(5)CFU/mL),followed by M-2(9.9×10^(5)CFU/mL)and control(4.4×10^(6)CFU/mL).In the culture water,20 isolates of culturable bacteria were found during the trial,with the dominant bacteria in M-2:Bacillus cereus,Bacillus velezensis,Gordonia polyisoprenivorans,T-2:Acinetobacter sp.,Gordonia polyisoprenivorans,Micrococcus luteus.Based on average well colour development(AWCD),the highest bacterial activity was in the M-2(0.297-0.707),followed by T-2(0.226-0.717)and control(0.094-0.694).The highest substrate group utilised in M-2 and T-2 were polymers(0.896 and 1.348)and control:amines(0.100).The highest Shannon diversity index was obtained in the M-2(2.508),followed by control(2.286)and the T-2(2.157).As many as 11 isolates of culturable bacteria were found in the shrimp intestine,with dominant bacteria in M-2:Bacillus velezensis,Micrococcus luteus,Bacillus cereus,T-2:Bacillus velezensis,Micrococcus sp.2,Demequina activiva.The highest bacterial activity was in M-2(2.259),followed by control(2.020)and T-2(0.050).The highest substrate group utilisation was in M-2:polymers(2.564),T-2:carbohydrates(0.094),and control:polymers(2.185).The highest Shannon diversity index was in the M-2(3.234),followed by control(3.230)and the T-2(0.751).In addition,1.5%M.circinelloides and T.harzianum supplementation increased white shrimp muscle protein content up to 79.21%and 80.40%,respectively,compared to control(76.73%).In conclusion,using Mucor circinelloides powder as a feed supplement can improve the growth performance of white shrimp,increase the functional/probiotic bacteria:B.cereus and B.velezensis in cultur and shrimp’s intestine,and increase the protein content in shrimp muscle tissue.However,more in-depth research is needed,especially on anti-nutritional compounds from fungi that affect shrimp growth.
基金Fundacao de Amparo a Pesquisa do Estado de Sao Paulo(Fapesp)and Conselho Nacional de Desenvolvimento Cientifico e Tecnologico(CNPq)for financial support
文摘The cultivation of the marine species of shrimp Litopenaeus vannamei has emerged as one of the segments that best develop in the Brazilian aquaculture, representing the agribusiness that grew the most in recent years. Regarding the economic importance of the farming of this species in Brazil, further studies on genetic improvement were conducted. A F2 segregating population consisting of 192 samples for each G1 and G2 families, from crossing inbred lines was used for the studies performed in the present work. The genetic linkage analysis was based on polymorphic markers derived from nine AFLP (Amplified Fragment Length Polymorphism) primers. Fourteen genetic linkage groups including 103 segregating polymorphic markers were constructed covering350 cMfor G1 and four genetic linkage groups including 59 markers were constructed covering300 cMfor G2. Simple marker analyses were performed among individuals evaluated phenotypically, finding markers linked to genes that may be potentially important and useful to assess characteristics of economic importance for traits related to weight and the disease IMN (idiopathic muscle necrosis). The statistical model including markers explained major proportion of phenotypic characteristic weight in relation to disease incidence IMN.
基金Supported by the Scientific Research Foundation of Third Institute of Oceanography,SOA(No.2011018)the Natural Science Foundation of Fujian Province of China(No.2011J05079)+3 种基金the National Natural Science Foundation of China(No.31001125)the Major State Basic Research Development Program of China(973 Program)(No.2012CB114403)the National High Technology Research and Development Program of China(863 Program)(No.2012AA092205)the China Agriculture Research System-47
文摘The phosphatidylinositol 3-kinase(PI3K)-AKT pathway is involved in various cellular functions, including anti-apoptosis, protein synthesis, glucose metabolism and cell cycling. However, the role of the PI3K-AKT pathway in crustaceans remains unclear. In the present study, we cloned and characterized the AKT gene lvakt from Litopenaeus vannamei. The 511-residue LVAKT was highly conserved; contained a PH domain, a catalytic domain and a hydrophobic domain; and was highly expressed in the heart and gills of L. vannamei. We found, using Real-Time Quantitative PCR(Q-PCR) analysis, that lvakt was upregulated during early white spot syndrome virus(WSSV) infection. Moreover, the PI3K-specific inhibitor, LY294002, reduced viral gene transcription, implying that the PI3K-AKT pathway might be hijacked by WSSV. Our results therefore suggest that LVAKT may play an important role in the shrimp immune response against WSSV.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(Nos.2012AA10A404,2012AA092205)the General Program of National Natural Science Foundation of China(Nos.31272683,31072203)to Dr.LI Fuhua
文摘Anti-lipopolysaccharide factors （ALFs） are basic components of the crustacean immune system that defend against a range of pathogens. The cDNA sequence of a new ALF, designated nLvALF2, with an open reading flame encoding 132 amino acids was cloned. Its deduced amino acid sequence contained the conserved functional domain of ALFs, the LPS binding domain （LBD）. its genotnic sequence consisted of three exons and four introns, nLvALF2 was mainly expressed in the Oka organ and gills of shrimps. The transcriptional level of nLw4LF2 increased significantly after white spot syndrome virus （WSSV） infection, suggesting its important roles in protecting shrimps from WSSV. Single nucleotide polymorphisms （SNPs） were found in the genomic sequence of nLvALF2, of which 38 were analyzed for associations with the susceptibility/resistance of shrimps to WSSV. The loci g.2422 A〉G, g.2466 T〉C, and g.2529 G〉A were significantly associated with the resistance to WSSV （P〈0.05）. These SNP loci could be developed as markers for selection of WSSV-resistant varieties ofLitopenaeus vannamei.
