The early life stages of marine organisms are pivotal in shaping community dynamics and resource availability.In this study,we focused on Portunus trituberculatus,a crustacean integral to China's fisheries economy...The early life stages of marine organisms are pivotal in shaping community dynamics and resource availability.In this study,we focused on Portunus trituberculatus,a crustacean integral to China's fisheries economy,and examined the effect of sea surface temperature(SST)in its critical early life stages on subsequent yields.To analyze the correlation between SST in different larval stages and the corresponding yield of P.trituberculatus,we simulated the transport and distribution of larvae from 2014 to 2022 by employing circulation models and Lagrangian particle tracking experiments(LPTE).In the five years(2014,2015,2016,2019,and 2020),particles were transported in a northwestern direction and moved in the direction of low SST.The distribution of particles in the megalopa stage(M stage)were located in the region of the lower temperature.In 2017,2018,and 2021,the particles were transported in a northeastern direction but they did not move with the gradient of low SST in these years,and the particles in the last M stage were located in the region where the SST was at the peak of the time period.In 2022,the distribution was observed for most of the particles in the southwestern part of Zhejiang coast,a small part of them were transported in the northwestern direction and a small amount of particles was distributed offshore along the northern area of the Zhejiang coast.The correlations between the SST at each stage of larvae with the corresponding year's yield showed that the yield of P.trituberculatus decreased significantly(R=-0.772,P=0.015)with increasing SST at the M stage.This study preliminarily explains the correlation between SST at the larval stage and the yield of P.trituberculatus and provides essential information for scientific stock enhancement in the future.展开更多
Swimming crab(Portunus trituberculatus)eggs contain abundant phospholipids and polyunsaturated fatty acids with specific bioactivities.However,more studies on the functions of other nutrients are needed.Here,a novel s...Swimming crab(Portunus trituberculatus)eggs contain abundant phospholipids and polyunsaturated fatty acids with specific bioactivities.However,more studies on the functions of other nutrients are needed.Here,a novel sialoglycopeptide from P.trituberculatus eggs,which was termed as Pt-SGP,was isolated with enzymolysis.Pt-SGP was a 6480-Da pure sialoglycopeptide.O-glycan units existed between peptides and sugars.Pt-SGP was composed of 10.69%protein,71.43%hexose,and 11.27%Neu5Gc acid.Analyzed with HPLC,GC-MS,and NMR spectrometry,the carbohydrate chain of Pt-SGP was{(4GlcNAc1-2Man1-3)/(4GlcNAc1-2Man1-6)Man1-6}/{4Glc1-4}Glc1-4Glc1-3GlcNAc1.The MS spectrometry analysis result showed that the peptide chain was Val-Ala-Thr*-Val-Ser*-Leu-Pro-Arg.The results on mice showed Pt-SGP could reduce body weight gain by 18.13%,white adipose weight by 28.57%,serum TC by 22.32,serum TG by 9.46%,serum LDL-C by 27.69%,hepatic TC by 73.21%,and hepatic TG by 25.20%.Furthernore,it could increase serum HDL-C by 75.96%,mitigate adipocyte size and hepatic lipid accumulation in high fat diet-fed mice.Pt-SGP significantly down-regulated C/EBPαsignaling and up-regulated AMPKαsignaling in the epididymal fat tissues.The findings indicate that Pt-SGP exhibits notable anti-obesity properties,and can potentially become a promising dietary supplement for anti-obesity.展开更多
The agglutination characteristics of serum agglutinin from Portunus trituberculatus were studied in this paper. The results showed that the serum agglutinin had no agglutination to crucian carp, Chinese soft shell tur...The agglutination characteristics of serum agglutinin from Portunus trituberculatus were studied in this paper. The results showed that the serum agglutinin had no agglutination to crucian carp, Chinese soft shell turtle, grass carp, chicken or human group A, B or O blood cells, but had strong agglutination to blood cells of mice and rabbits. The activities of serum agglutinin from Portunus trituberculatus to mice blood cells reached 210, and to rabbit blood cells reached 28. Salinity had a greater effect on agglutinin activity. When the Na Cl concentration exceeded 0.6 mol/L,the serum agglutinin from Portunus trituberculatus was basically inactivated. The optimum p H for agglutinin activity was 6.0-7.4. The serum agglutinin from Portunus trituberculatus had obvious dependence to Ca2 ＋and Mg2 ＋, and EDTA could significantly inhibit its activity. The results of sugar inhibition test showed that the activity of agglutinin from Portunus trituberculatus can be specifically inhibited by N-acetylglucosamine and N-acetylmannosamine. The serum agglutinin from Portunus trituberculatus was isolated by ammonium sulfate gradient precipitation, and its activity was highest by the 25% ammonium sulfate precipitation system. The SDS polyacrylamide gel electrophoresis（SDS-PAGE） showed that the protein bands were mainly distributed within 72-95 ku.展开更多
As an important commercial species, the blue swimming crabs with different body-colors were discovered along the coast of East China in recent years. Both purple and tea-green crabs have similar, even the same physica...As an important commercial species, the blue swimming crabs with different body-colors were discovered along the coast of East China in recent years. Both purple and tea-green crabs have similar, even the same physical characteristics except for the carapace color. In order to identify the relationships between them, we amplified and compared two conserved mitochondrial gene fragments of color-different crab individuals through PCR-SSCP and DNA sequencing analyses. The results showed that there was no distinct variation of DNA bands of both the COl and 16srRNA genes detected in polyacrylamide gels through SSCP analysis between purple and tea-green crab samples, and the following sequence analysis of color-different crab individuals presented 99.81% and 99.91% nucleotide sequence identity of the COl and 16srRNA genes respectively, which indicated that there was no species or subspecies differentiation between them. In addition, the purple individuals were more conserved in DNA sequence in compadson with that of the tee-green ones.展开更多
The blue swimming crabs Portunus trituberculatus with tea-green and/or purple carapace color have been discovered on the shore of eastern part of China these days, In this paper, 40 color-different crab individuals we...The blue swimming crabs Portunus trituberculatus with tea-green and/or purple carapace color have been discovered on the shore of eastern part of China these days, In this paper, 40 color-different crab individuals were evaluated using 80 RAPD primers. 30 primers were selected using bulk segregation analysis （BSA） and RAPD-PCR analysis. The results showed that the ratio of polymorphic loci was 61.29%, and Nei＇s gene diversity was 0.375 6, and Shannon＇s information index was 0.554 5 among purple crab individuals; and the ratio of polymorphic loci was 62.44%, and Nei＇s gene diversity was 0.3451, and Shannon＇s information index was 0.5099 among tea-green crab individuals. It was clear that the genetic diversity between purple and tea-green crabs was similar, which indicated close phylogenetic relationships between the purple and tea-green crabs. In addition, two primers （SBS A09 and SBS G16） got specific DNA bands by comparing forty individuals of color-different crab individuals through RAPD-PCR analysis and the following electrophoresis. They indicated close phylogenetic relationships between purple and tea-green crabs.展开更多
This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of t...This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.展开更多
Na^＋/K^＋-ATPases are membrane-associated enzymes responsible for the active transport of Na^＋ and K^＋ ions across cell membranes, generating chemical and electrical gradients. These enzymes＇ α-subunit provides c...Na^＋/K^＋-ATPases are membrane-associated enzymes responsible for the active transport of Na^＋ and K^＋ ions across cell membranes, generating chemical and electrical gradients. These enzymes＇ α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^＋/K^＋-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs （bp）, with an open reading frame of 3 120 bp, 5＇ untranslated region （UTR） of 317 bp, and 3＇ UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^＋/K^＋-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit＇s transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab＇s Na^＋/K^＋-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.展开更多
In the survey of fishery resources,the sampling design will directly impact the accuracy of the estimation of the abundance.Therefore,it is necessary to optimize the sampling design to increase the quality of fishery ...In the survey of fishery resources,the sampling design will directly impact the accuracy of the estimation of the abundance.Therefore,it is necessary to optimize the sampling design to increase the quality of fishery surveys.The distribution and abundance of fisheries resource estimated based on the bottom trawl survey data in the Changjiang River(Yangtze River)Estuary-Hangzhou Bay and its adjacent waters in 2007 were used to simulate the"true"situation.Then the abundance index of Portunus trituberculatus were calculated and compared with its true index to evaluate the impacts of different sampling designs on the abundance estimation.Four sampling methods(including fixed-station sampling,simple random sampling,stratified fixed-station sampling,and stratified random sampling)were simulated.Three numbers of stations(9,16 and 24)were assumed for the scenarios of fixed-station sampling and simple random sampling without stratification.While 16 stations were assumed for the scenarios with stratification.Three reaction distances(1.5 m,3 m and 5 m)of P.trituberculatus to the bottom line of trawl were also assumed to adapt to the movement ability of the P.trituberculatus for different ages,seasons and substrate conditions.Generally speaking,compared with unstratified sampling design,the stratified sampling design resulted in more accurate abundance estimation of P.trituberculatus,and simple random sampling design is better than fixed-station sampling design.The accuracy of the simulated results was improved with the increase of the station number.The maximum relative estimation error(REE)was 163.43%and the minimum was 49.40%for the fixed-station sampling scenario with 9 stations,while 38.62%and 4.15%for 24 stations.With the increase of reaction distance,the relative absolute bias(RAB)and REE gradually decreased.Resource-intensive area and the seasons with high density variances have significant impacts on simulation results.Thus,it will be helpful if there are prior information or pre-survey results about density distribution.The current study can provide reference for the future sampling design of bottom trawl of P.trituberculatus and other species.展开更多
This study was conducted to evaluate the effects of dietary lipid sources on the growth performance and fatty acid composition of the swimming crab, P ortunus trituberculatus. Four isonitrogenous and isoenergetic expe...This study was conducted to evaluate the effects of dietary lipid sources on the growth performance and fatty acid composition of the swimming crab, P ortunus trituberculatus. Four isonitrogenous and isoenergetic experimental diets were formulated to contain four separate lipid sources, including fish, soybean, rapeseed, and linseed oils(FO, SO, RO, and LO, respectively). With three replicates of 18 crabs each for each diet, crabs(initial body weight, 17.00 ±0.09 g) were fed twice daily for 8 weeks. There were no significant differences among these groups in terms of weight gain, specific growth rate, and hepatosomatic index. However, the RO groups' survival rate was significantly lower than FO groups. The feed conversion and protein efficiency ratios of RO groups were poorer than other groups. The proximate compositions of whole body and hepatopancreas were significantly affected by these dietary treatments. Tissue fatty acid composition mainly reflected dietary fatty acid compositions. Crabs fed FO diets exhibited significantly higher arachidonic, eicosapentaenoic, and docosahexaenoic acid contents in muscle and hepatopancreas compared with VO crabs. Linoleic, oleic, and linolenic acids in muscle and hepatopancreas were the highest in the SO, RO, and LO groups, respectively. The present study suggested that SO and LO could substitute for FO in fishmeal-based diets for swimming crabs, without affecting growth performance and survival.展开更多
A bstract Gut microbiota impacts the health of crustaceans. V ibrio alginolyticus is a main causative pathogen that induces the vibriosis in farmed swimming crabs, Portunus trituberculatus. However, it remains unknown...A bstract Gut microbiota impacts the health of crustaceans. V ibrio alginolyticus is a main causative pathogen that induces the vibriosis in farmed swimming crabs, Portunus trituberculatus. However, it remains unknown whether gut bacteria perform functions during the progression of vibriosis. In this study, 16 SrRNA gene amplicon sequencing was used to investigate temporal alteration of gut bacterial community in swimming crabs in response to 72-h V. alginolyticus challenge. Our results show that V. alginolyticus infection resulted in dynamic changes of bacterial community composition in swimming crabs. Such changes were highlighted by the overwhelming overabundance of V ibrio and a significant fluctuation in the gut bacteria including the bacteria with high relative abundance and especially those with low relative abundance. These findings reveal that crab vibriosis gradually develops with the infection time of V. alginolyticus and tightly relates to the dysbiosis of gut bacterial community structure. This work contributes to our appreciation of the importance of the balance of gut bacterial community structure in maintaining the health of crustaceans.展开更多
Heavy metals,polycyclic aromatic hydrocarbons(PAHs),and perfluorinated alkyl substances(PFASs)polluted in 155 samples of two marine crustaceans(Oratosquilla oratoria and Portunus trituberculatus)harvested in the Zhous...Heavy metals,polycyclic aromatic hydrocarbons(PAHs),and perfluorinated alkyl substances(PFASs)polluted in 155 samples of two marine crustaceans(Oratosquilla oratoria and Portunus trituberculatus)harvested in the Zhoushan fishing ground in the East China Sea were assessed.The results showed that Pb,Hg,and iAs in the whole edible parts of O.oratoria and P.trituberculatus were at trace levels,far below the limits set by the China’s national standard,whereas the exceeding standard rates of Cd in the whole edible parts of O.oratoria and P.trituberculatus were 98.6%and 75.6%,respectively.Moreover,the average Cd levels and the exceeding standard rates of Cd in the visceral tissues of O.oratoria were significantly higher than those in the muscle tissues(4.64 mg kg^(−1)vs.0.49 mg kg^(−1)and 96.7%vs 30%,respectively).PAHs with high detection frequency in the two marine crustaceans included NA,AC,FL,and PHE.Total contents of 15 PAHs(abbreviated asΣ15 PAHs)in whole edible parts of O.oratoria and P.trituberculatus were 10.75–65.9μg kg^(−1)(mean=18.7μg kg^(−1)and 13.26–181μg kg^(−1)(mean=24.2μg kg^(−1),and those in the muscle and the visceral tissues of P.trituberculatus were 9.58–160μg kg^(−1)(mean=21.1μg kg^(−1)and 18.22–201μg kg^(−1)(mean=32.7μg kg^(−1),respectively.Furthermore,PFASs that were found at high contents included PFOA,PFOS,and PFOSA,and total contents of 23 PFASs(Σ23 PFASs)ranged 0.0162–5.104μg kg^(−1)(mean=0.915μg kg^(−1).Collectively,the present work provides new data on the contamination in two marine crustaceans captured in the Zhoushan fishing ground in the China East Sea,which can be useful for making suggestions on proper consumption amounts of the two marine crustaceans.展开更多
Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also in...Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ^(13)C value for all the biomass groups in polyculture ecosystem ranged from -25.61‰ to -16.60‰, and the mean δ^(15)N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ^(13)C value was found between particulate organic matter(POM) and sediment organic matter(SOM)(P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ^(13)C-enrichment factor of 1.00‰ and a ^(15)N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.展开更多
A mass mortality often occurs from molting to the megalopa stage during the larval development of the swimming crab Portunus trituberculatus. Larvae with insuffcient nutrient accumulation during the zoeal stages are p...A mass mortality often occurs from molting to the megalopa stage during the larval development of the swimming crab Portunus trituberculatus. Larvae with insuffcient nutrient accumulation during the zoeal stages are probably unable to develop into juvenile swimming crabs. However, the nutritional information such as the primary metabolites is scarce for P. trituberculatus larvae. The aim of this work is to obtain an insight into the metabolite traits of P. trituberculatus at early developmental stages. 1 H nuclear magnetic resonance spectroscopy coupled with multivariate data analysis was used to determine how the metabolite pro?les shift during larval development in P. trituberculatus. Our results show that the trend of total metabolites exhibited a rise from zoea 1 to zoea 3, followed by a drop from zoea 4 to megalopa and recovery during the ?rst juvenile stage. A large-scale depletion of total metabolites in the zoea 4 and megalopa stages suggests a deep depression of metabolic activity, which may be linked to the mass mortality from molting to the megalopa stage. These ?ndings provided essential metabolic information about the larval development of P. trituberculatus and important clues for understanding the nutritional requirements of P. trituberculatus in early developmental stages.展开更多
We established a line (screened) of Portunus trituberculatus by selectively breeding individuals that survived from challenge with Vibrio alginolyticus, and compared the response of screened and unscreened (control) P...We established a line (screened) of Portunus trituberculatus by selectively breeding individuals that survived from challenge with Vibrio alginolyticus, and compared the response of screened and unscreened (control) P. trituberculatus challenged with V. alginolyticus. We measured superoxide dismutase, catalase, acid phosphatase, alkaline phosphatase, and peroxidase activity and the content of hemocyanin in the plasma and phenoloxidase activity in serum. The cumulative survival rate after 24-h challenge with V. alginolyticus was significantly higher in the screened crabs than in the unscreened crabs (P<0.05). T-SOD and PO activity were significantly lower in the screened stock than in the unscreened stock (P<0.05). POD, CAT, and ACP activity and hemocyanin content were significantly higher in the screened stock than in the unscreened stock. Our results suggest that the screened stock was more resistant to infection. Furthermore, the indices we measured may be used to evaluate the health state of P. trituberculatus.展开更多
基金Supported by the National Key Research and Development Program of China(No.2019YFD0901304)the Public Welfare Technology Application Research Project of Zhejiang(No.LGN21C190009)the Science and Technology Project of Zhoushan(No.2022C41003)。
文摘The early life stages of marine organisms are pivotal in shaping community dynamics and resource availability.In this study,we focused on Portunus trituberculatus,a crustacean integral to China's fisheries economy,and examined the effect of sea surface temperature(SST)in its critical early life stages on subsequent yields.To analyze the correlation between SST in different larval stages and the corresponding yield of P.trituberculatus,we simulated the transport and distribution of larvae from 2014 to 2022 by employing circulation models and Lagrangian particle tracking experiments(LPTE).In the five years(2014,2015,2016,2019,and 2020),particles were transported in a northwestern direction and moved in the direction of low SST.The distribution of particles in the megalopa stage(M stage)were located in the region of the lower temperature.In 2017,2018,and 2021,the particles were transported in a northeastern direction but they did not move with the gradient of low SST in these years,and the particles in the last M stage were located in the region where the SST was at the peak of the time period.In 2022,the distribution was observed for most of the particles in the southwestern part of Zhejiang coast,a small part of them were transported in the northwestern direction and a small amount of particles was distributed offshore along the northern area of the Zhejiang coast.The correlations between the SST at each stage of larvae with the corresponding year's yield showed that the yield of P.trituberculatus decreased significantly(R=-0.772,P=0.015)with increasing SST at the M stage.This study preliminarily explains the correlation between SST at the larval stage and the yield of P.trituberculatus and provides essential information for scientific stock enhancement in the future.
基金supported by the National Natural Science Foundation of China(No.41806182)the National Key R&D Program of China(No.2020YFD0900900)+1 种基金the Provincial Universities Basic Scientific Research Foundation in Zhejiang Province of China(No.2021ZJ004)the Science and Technology Project of Zhoushan(No.2020C21020).
文摘Swimming crab(Portunus trituberculatus)eggs contain abundant phospholipids and polyunsaturated fatty acids with specific bioactivities.However,more studies on the functions of other nutrients are needed.Here,a novel sialoglycopeptide from P.trituberculatus eggs,which was termed as Pt-SGP,was isolated with enzymolysis.Pt-SGP was a 6480-Da pure sialoglycopeptide.O-glycan units existed between peptides and sugars.Pt-SGP was composed of 10.69%protein,71.43%hexose,and 11.27%Neu5Gc acid.Analyzed with HPLC,GC-MS,and NMR spectrometry,the carbohydrate chain of Pt-SGP was{(4GlcNAc1-2Man1-3)/(4GlcNAc1-2Man1-6)Man1-6}/{4Glc1-4}Glc1-4Glc1-3GlcNAc1.The MS spectrometry analysis result showed that the peptide chain was Val-Ala-Thr*-Val-Ser*-Leu-Pro-Arg.The results on mice showed Pt-SGP could reduce body weight gain by 18.13%,white adipose weight by 28.57%,serum TC by 22.32,serum TG by 9.46%,serum LDL-C by 27.69%,hepatic TC by 73.21%,and hepatic TG by 25.20%.Furthernore,it could increase serum HDL-C by 75.96%,mitigate adipocyte size and hepatic lipid accumulation in high fat diet-fed mice.Pt-SGP significantly down-regulated C/EBPαsignaling and up-regulated AMPKαsignaling in the epididymal fat tissues.The findings indicate that Pt-SGP exhibits notable anti-obesity properties,and can potentially become a promising dietary supplement for anti-obesity.
基金Supported by National Natural Science Foundation of China(41106123)National High Technology Research and Development Program of China(863 Program)(2012AA10A409)+1 种基金Natural Science Foundation of Zhejiang Province(LY12C19009)Key Special Project of Department of Science and Technology of Zhejiang Province(2012C12907-3)
文摘The agglutination characteristics of serum agglutinin from Portunus trituberculatus were studied in this paper. The results showed that the serum agglutinin had no agglutination to crucian carp, Chinese soft shell turtle, grass carp, chicken or human group A, B or O blood cells, but had strong agglutination to blood cells of mice and rabbits. The activities of serum agglutinin from Portunus trituberculatus to mice blood cells reached 210, and to rabbit blood cells reached 28. Salinity had a greater effect on agglutinin activity. When the Na Cl concentration exceeded 0.6 mol/L,the serum agglutinin from Portunus trituberculatus was basically inactivated. The optimum p H for agglutinin activity was 6.0-7.4. The serum agglutinin from Portunus trituberculatus had obvious dependence to Ca2 ＋and Mg2 ＋, and EDTA could significantly inhibit its activity. The results of sugar inhibition test showed that the activity of agglutinin from Portunus trituberculatus can be specifically inhibited by N-acetylglucosamine and N-acetylmannosamine. The serum agglutinin from Portunus trituberculatus was isolated by ammonium sulfate gradient precipitation, and its activity was highest by the 25% ammonium sulfate precipitation system. The SDS polyacrylamide gel electrophoresis（SDS-PAGE） showed that the protein bands were mainly distributed within 72-95 ku.
基金supported by the grant of the Key Project in the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (2006BAD09A01)
文摘As an important commercial species, the blue swimming crabs with different body-colors were discovered along the coast of East China in recent years. Both purple and tea-green crabs have similar, even the same physical characteristics except for the carapace color. In order to identify the relationships between them, we amplified and compared two conserved mitochondrial gene fragments of color-different crab individuals through PCR-SSCP and DNA sequencing analyses. The results showed that there was no distinct variation of DNA bands of both the COl and 16srRNA genes detected in polyacrylamide gels through SSCP analysis between purple and tea-green crab samples, and the following sequence analysis of color-different crab individuals presented 99.81% and 99.91% nucleotide sequence identity of the COl and 16srRNA genes respectively, which indicated that there was no species or subspecies differentiation between them. In addition, the purple individuals were more conserved in DNA sequence in compadson with that of the tee-green ones.
基金supported by the grant of the Key Project in the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (No. 2006BAD09A01)
文摘The blue swimming crabs Portunus trituberculatus with tea-green and/or purple carapace color have been discovered on the shore of eastern part of China these days, In this paper, 40 color-different crab individuals were evaluated using 80 RAPD primers. 30 primers were selected using bulk segregation analysis （BSA） and RAPD-PCR analysis. The results showed that the ratio of polymorphic loci was 61.29%, and Nei＇s gene diversity was 0.375 6, and Shannon＇s information index was 0.554 5 among purple crab individuals; and the ratio of polymorphic loci was 62.44%, and Nei＇s gene diversity was 0.3451, and Shannon＇s information index was 0.5099 among tea-green crab individuals. It was clear that the genetic diversity between purple and tea-green crabs was similar, which indicated close phylogenetic relationships between the purple and tea-green crabs. In addition, two primers （SBS A09 and SBS G16） got specific DNA bands by comparing forty individuals of color-different crab individuals through RAPD-PCR analysis and the following electrophoresis. They indicated close phylogenetic relationships between purple and tea-green crabs.
基金supported by the Natural Science Foundation of Shandong Province, China (No. ZR2016 CM21)
文摘This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A409)the National Natural Science Foundation of China(No.41306177)the Special Scientific Research Funds for Central Non-Profit Institutes,Yellow Sea Fisheries Research Institutes(No.20603022013027)
文摘Na^＋/K^＋-ATPases are membrane-associated enzymes responsible for the active transport of Na^＋ and K^＋ ions across cell membranes, generating chemical and electrical gradients. These enzymes＇ α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^＋/K^＋-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs （bp）, with an open reading frame of 3 120 bp, 5＇ untranslated region （UTR） of 317 bp, and 3＇ UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^＋/K^＋-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit＇s transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab＇s Na^＋/K^＋-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.
基金The National Key Research and Development Program of China under contract No.2017YFA0604902the Science and Technology Project of Zhoushan under contract No.2017C41012。
文摘In the survey of fishery resources,the sampling design will directly impact the accuracy of the estimation of the abundance.Therefore,it is necessary to optimize the sampling design to increase the quality of fishery surveys.The distribution and abundance of fisheries resource estimated based on the bottom trawl survey data in the Changjiang River(Yangtze River)Estuary-Hangzhou Bay and its adjacent waters in 2007 were used to simulate the"true"situation.Then the abundance index of Portunus trituberculatus were calculated and compared with its true index to evaluate the impacts of different sampling designs on the abundance estimation.Four sampling methods(including fixed-station sampling,simple random sampling,stratified fixed-station sampling,and stratified random sampling)were simulated.Three numbers of stations(9,16 and 24)were assumed for the scenarios of fixed-station sampling and simple random sampling without stratification.While 16 stations were assumed for the scenarios with stratification.Three reaction distances(1.5 m,3 m and 5 m)of P.trituberculatus to the bottom line of trawl were also assumed to adapt to the movement ability of the P.trituberculatus for different ages,seasons and substrate conditions.Generally speaking,compared with unstratified sampling design,the stratified sampling design resulted in more accurate abundance estimation of P.trituberculatus,and simple random sampling design is better than fixed-station sampling design.The accuracy of the simulated results was improved with the increase of the station number.The maximum relative estimation error(REE)was 163.43%and the minimum was 49.40%for the fixed-station sampling scenario with 9 stations,while 38.62%and 4.15%for 24 stations.With the increase of reaction distance,the relative absolute bias(RAB)and REE gradually decreased.Resource-intensive area and the seasons with high density variances have significant impacts on simulation results.Thus,it will be helpful if there are prior information or pre-survey results about density distribution.The current study can provide reference for the future sampling design of bottom trawl of P.trituberculatus and other species.
基金Supported by the Agriculture Science Technology Achievement Transformation Fund(No.2014GB2C22015)the Public Projects of Zhejiang Province(Nos.2013C32037,2013C31032)+3 种基金the Zhejiang Major Special Program of Breeding(No.2012C12907-3)the Ningbo Innovation and Entrepreneurship Project(No.2014C92011)the Zhejiang Provincial Oceanic and Fishery Bureau ProjectZhoushan Science and Technology Bureau Project(No.2013C41007)
文摘This study was conducted to evaluate the effects of dietary lipid sources on the growth performance and fatty acid composition of the swimming crab, P ortunus trituberculatus. Four isonitrogenous and isoenergetic experimental diets were formulated to contain four separate lipid sources, including fish, soybean, rapeseed, and linseed oils(FO, SO, RO, and LO, respectively). With three replicates of 18 crabs each for each diet, crabs(initial body weight, 17.00 ±0.09 g) were fed twice daily for 8 weeks. There were no significant differences among these groups in terms of weight gain, specific growth rate, and hepatosomatic index. However, the RO groups' survival rate was significantly lower than FO groups. The feed conversion and protein efficiency ratios of RO groups were poorer than other groups. The proximate compositions of whole body and hepatopancreas were significantly affected by these dietary treatments. Tissue fatty acid composition mainly reflected dietary fatty acid compositions. Crabs fed FO diets exhibited significantly higher arachidonic, eicosapentaenoic, and docosahexaenoic acid contents in muscle and hepatopancreas compared with VO crabs. Linoleic, oleic, and linolenic acids in muscle and hepatopancreas were the highest in the SO, RO, and LO groups, respectively. The present study suggested that SO and LO could substitute for FO in fishmeal-based diets for swimming crabs, without affecting growth performance and survival.
基金Supported by the National Natural Science Foundation of China(No.41673076)the Major Agriculture Program of Ningbo(No.2017C110007)the K.C.Wong Magna Fund in Ningbo University
文摘A bstract Gut microbiota impacts the health of crustaceans. V ibrio alginolyticus is a main causative pathogen that induces the vibriosis in farmed swimming crabs, Portunus trituberculatus. However, it remains unknown whether gut bacteria perform functions during the progression of vibriosis. In this study, 16 SrRNA gene amplicon sequencing was used to investigate temporal alteration of gut bacterial community in swimming crabs in response to 72-h V. alginolyticus challenge. Our results show that V. alginolyticus infection resulted in dynamic changes of bacterial community composition in swimming crabs. Such changes were highlighted by the overwhelming overabundance of V ibrio and a significant fluctuation in the gut bacteria including the bacteria with high relative abundance and especially those with low relative abundance. These findings reveal that crab vibriosis gradually develops with the infection time of V. alginolyticus and tightly relates to the dysbiosis of gut bacterial community structure. This work contributes to our appreciation of the importance of the balance of gut bacterial community structure in maintaining the health of crustaceans.
基金the National Key R&D Program of China(No.2020YFD0900900)the Public Interest Science and Technology Plan of Ningbo(No.2019C10007)the Basic Public Welfare Project of Zhejiang Provincial Department of Science and Technol ogy(No.LGN20C200015).
文摘Heavy metals,polycyclic aromatic hydrocarbons(PAHs),and perfluorinated alkyl substances(PFASs)polluted in 155 samples of two marine crustaceans(Oratosquilla oratoria and Portunus trituberculatus)harvested in the Zhoushan fishing ground in the East China Sea were assessed.The results showed that Pb,Hg,and iAs in the whole edible parts of O.oratoria and P.trituberculatus were at trace levels,far below the limits set by the China’s national standard,whereas the exceeding standard rates of Cd in the whole edible parts of O.oratoria and P.trituberculatus were 98.6%and 75.6%,respectively.Moreover,the average Cd levels and the exceeding standard rates of Cd in the visceral tissues of O.oratoria were significantly higher than those in the muscle tissues(4.64 mg kg^(−1)vs.0.49 mg kg^(−1)and 96.7%vs 30%,respectively).PAHs with high detection frequency in the two marine crustaceans included NA,AC,FL,and PHE.Total contents of 15 PAHs(abbreviated asΣ15 PAHs)in whole edible parts of O.oratoria and P.trituberculatus were 10.75–65.9μg kg^(−1)(mean=18.7μg kg^(−1)and 13.26–181μg kg^(−1)(mean=24.2μg kg^(−1),and those in the muscle and the visceral tissues of P.trituberculatus were 9.58–160μg kg^(−1)(mean=21.1μg kg^(−1)and 18.22–201μg kg^(−1)(mean=32.7μg kg^(−1),respectively.Furthermore,PFASs that were found at high contents included PFOA,PFOS,and PFOSA,and total contents of 23 PFASs(Σ23 PFASs)ranged 0.0162–5.104μg kg^(−1)(mean=0.915μg kg^(−1).Collectively,the present work provides new data on the contamination in two marine crustaceans captured in the Zhoushan fishing ground in the China East Sea,which can be useful for making suggestions on proper consumption amounts of the two marine crustaceans.
基金supported by the National Great Project of Scientific and Technical Supporting Programs(No.2011BAD13B03)the Programs for Excellent Youth Foundation of Shandong Province(No.JQ201009)
文摘Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ^(13)C value for all the biomass groups in polyculture ecosystem ranged from -25.61‰ to -16.60‰, and the mean δ^(15)N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ^(13)C value was found between particulate organic matter(POM) and sediment organic matter(SOM)(P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ^(13)C-enrichment factor of 1.00‰ and a ^(15)N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.
基金Supported by the National Natural Science Foundation of China(No.41673076)the Major Agriculture Program of Ningbo(No.2017C110007)+1 种基金China Agriculture Research System-CARS48the K.C.Wong Magna Fund in Ningbo University
文摘A mass mortality often occurs from molting to the megalopa stage during the larval development of the swimming crab Portunus trituberculatus. Larvae with insuffcient nutrient accumulation during the zoeal stages are probably unable to develop into juvenile swimming crabs. However, the nutritional information such as the primary metabolites is scarce for P. trituberculatus larvae. The aim of this work is to obtain an insight into the metabolite traits of P. trituberculatus at early developmental stages. 1 H nuclear magnetic resonance spectroscopy coupled with multivariate data analysis was used to determine how the metabolite pro?les shift during larval development in P. trituberculatus. Our results show that the trend of total metabolites exhibited a rise from zoea 1 to zoea 3, followed by a drop from zoea 4 to megalopa and recovery during the ?rst juvenile stage. A large-scale depletion of total metabolites in the zoea 4 and megalopa stages suggests a deep depression of metabolic activity, which may be linked to the mass mortality from molting to the megalopa stage. These ?ndings provided essential metabolic information about the larval development of P. trituberculatus and important clues for understanding the nutritional requirements of P. trituberculatus in early developmental stages.
基金Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2012AA10A409)the National Natural Science Foundation of China (No. 41106123)+10 种基金the Transformation Fund for Agricultural Science (No. 2010GB2C220537)the Research Fund for the Doctoral Program of Higher Education (No. 20103305120002)the Scientific Research Fund of Zhejiang Provincial Education Department (No.Z201121258)the Zhejiang Key Innovative Team of Marine Aquaculture(No. 2010R50025-08)the Ningbo Natural Science Foundation (No.2010A610004)the Ningbo Innovative Program of Agriculture (No.2011C92014)the Ningbo Special Program of Agriculture (No. 201015)the Ningbo Special Program of Marine Fishing (No. 2011-1-7)the Public Interest Program of Zhejiang Province (No. 2011C33025)the Program for Science and Technology Innovative Research Team of Ningbo (No.2011B81003)the K C Wong Magana Fund in Ningbo University
文摘We established a line (screened) of Portunus trituberculatus by selectively breeding individuals that survived from challenge with Vibrio alginolyticus, and compared the response of screened and unscreened (control) P. trituberculatus challenged with V. alginolyticus. We measured superoxide dismutase, catalase, acid phosphatase, alkaline phosphatase, and peroxidase activity and the content of hemocyanin in the plasma and phenoloxidase activity in serum. The cumulative survival rate after 24-h challenge with V. alginolyticus was significantly higher in the screened crabs than in the unscreened crabs (P<0.05). T-SOD and PO activity were significantly lower in the screened stock than in the unscreened stock (P<0.05). POD, CAT, and ACP activity and hemocyanin content were significantly higher in the screened stock than in the unscreened stock. Our results suggest that the screened stock was more resistant to infection. Furthermore, the indices we measured may be used to evaluate the health state of P. trituberculatus.
