Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficienc...Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficiency is limited. For this purpose, the embryonic and larval development of reciprocal crosses of Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) were studied under laboratory conditions. The fertilization rate varied from 63.33% to 92%, while the hatching rate ranged from 55.68% to 76% with the highest value in hybrids ♀Cg × ♂Cj. Crosses between ♀Cj × ♂Cj, ♀Cg × ♂Cj and ♀Cj × ♂Cg had embryonic stages similar to those of the pure sib ♀Cg x ♂Cg. All crosses, however, had different timing for the various embryological stages. Hatching occurred at 32 h 15 min and 38 h for ♀Cj × ♂Cj and ♀Cj × ♂Cg, and 23 h and 23 h 30 min, respectively, for ♀Cg × ♂Cg and ♀Cg × ♂Cj. However, both crosses produced viable larvae until the first external feeding. The external morphological features of the larvae were completely formed by the 10th day after hatching. The body forms of the crosses at this time were indistinguishable from the pure sib. This study thus laid the groundwork for further comparative studies on hybrid performance and characterization.展开更多
Bacterial community presumably plays an essential role in inhibiting pathogen colonization and maintaining the health of scallop larvae, but limiting data are available for Yesso scallop （Patinopecten yessoensisis Ja...Bacterial community presumably plays an essential role in inhibiting pathogen colonization and maintaining the health of scallop larvae, but limiting data are available for Yesso scallop （Patinopecten yessoensisis Jay, 1857） larval development stages. The aim of this study was to characterize and compare the bacterial communities associating with Yesso scallop larval development at fertilized egg S l, trochophora S2, D-shaped larvae S3, umbo larvae S4, and juvenile scallop S5 stages by Illumina high-throughput sequencing. Genomic DNA was extracted from the larvae and their associating baetera, and a gene segment covering V3-V4 region of 16S rRNA gene was amplified and sequenced using an Illumina Miseq sequencer. Overall, 106760 qualified sequences with an average length of 449 bp were obtained. Sequences were compared with those retrieved from 16S rRNA gene databases, and 4 phyla, 7 classes, 15 orders, 21 families, 31 genera were identified. Proteobacteria was predominant phylum, accounting for more than 99%, at all 5 larval development stages. At genus level, Pseudomonas was dominant at stages S1 （80.60%）, S2 （87.77%） and S5 （68.71%）, followed by Photobacterium （17.06%） and Aeromonas （1.64%） at stage S1, Serratia （6.94%）, Stenotrophomonas （3.08%） and Acinetobacter （1.2%） at stage S2, Shewanella （25.95%） and Pseudoalteromonas （4.57%） at stage S5. Moreover, genus Pseudoal- teromonas became dominant at stages S3 （44.85%） and S4 （56.02%）, followed by Photobacterium （29.82%）, Pseudomonas （11.86%）, Aliivibrio （8.60%） and Shewanella （3.39%） at stage S3, Pseudomonas （18.16%）, Aliivibrio （14.29%）, Shewanella （4.11%）, Psychro- monas （4.04%） and Psychrobacter （1.81%） at stage S4. From the results, we concluded that the bacterial community changed sig- nificantly at different development stages of Yesso Scallop larvae.展开更多
We evaluated the effect of pH on larval development in larval Pacific oyster(Crassostrea gigas) and blood cockle(Arca inflata Reeve).The larvae were reared at pH 8.2(control),7.9,7.6,or 7.3beginning 30 min or 24 h pos...We evaluated the effect of pH on larval development in larval Pacific oyster(Crassostrea gigas) and blood cockle(Arca inflata Reeve).The larvae were reared at pH 8.2(control),7.9,7.6,or 7.3beginning 30 min or 24 h post fertilization.Exposure to lower pH during early embryonic development inhibited larval shell formation in both species.Compared with the control,larvae took longer to reach the D-veliger stage when reared under pH 7.6 and 7.3.Exposure to lower pH immediately after fertilization resulted in significantly delayed shell formation in the Pacific oyster larvae at pH 7.3 and blood cockle larvae at pH 7.6 and 7.3.However,when exposure was delayed until 24 h post fertilization,shell formation was only inhibited in blood cockle larvae reared at pH 7.3.Thus,the early embryonic stages were more sensitive to acidified conditions.Our results suggest that ocean acidification will have an adverse effect on embryonic development in bivalves.Although the effects appear subtle,they may accumulate and lead to subsequent issues during later larval development.展开更多
The swimming crab Portunus trituberculatus is an economically important marine crustacean species.Here isobaric tags for relative and absolute quantitation(iTRAQ)analysis were used to identify proteins that are differ...The swimming crab Portunus trituberculatus is an economically important marine crustacean species.Here isobaric tags for relative and absolute quantitation(iTRAQ)analysis were used to identify proteins that are differentially expressed during larval development of P.trituberculatus to elucidate the underlying mechanisms.In comparison with the first zoea stage(Z1),3980 proteins were identified from 32789 peptides,which were matched with 115497 spectrums.A total of 241 proteins were screened with significantly differential expressions in all development stages.These 241 proteins are involved in various biological processes,such as cytoskeleton organization,protein synthesis,energy production and substance metabolism,physiological activities,and transport.Cluster analyses of the 241 differentially expressed proteins led to the generation of four protein clusters based on the overall similarity in protein expression patterns.Exactly 54,70,36,and 45 proteins clustered in profiles 10(0,0,0,−1,0,0),15(0,1,0,1,0,1),18(0,1,2,2,1,0),and 19(0,1,2,3,4,5),respectively.Muscle development and exoskeleton renewal were important processes throughout the development stages.In addition,protein synthesis,degradation,and digestion actively occurred,especially at the Z4 stage.These results provide novel insights into the mechanisms underlying larval development of swimming crab and can assist in larval rearing.展开更多
With increasing demand in China for the mitten crab larvae, understanding its survival mechanism gets more important. This research focused on the effects of temperature and Ca 2+ on the larval growth and development....With increasing demand in China for the mitten crab larvae, understanding its survival mechanism gets more important. This research focused on the effects of temperature and Ca 2+ on the larval growth and development. Eriocheir sinensis larvae were reared in laboratory under 21 different combinations of temperature (15, 20, 25℃) and Ca 2+ content (120, 130, 140, 150, 160, 170, 180 mg/l) and constant salinity (20) and pH (8). The results suggested that the survival rate increases with temperature and Ca 2+ content. These combinations of temperature and Ca 2+ content maximized survival rate in our study and it may be the optimum water environmental conditions for culturing the larvae. To predict surviving larvae number under different water environmental conditions, 21 dynamic mathematical models were developed. This for the first time observation of the zoeal Ⅵ larvae of the Changjiang River E. sinensis population showed that they occurred under stressed water environmental conditions: temperature of 15℃ and Ca 2+ content of 120,130 mg/l.展开更多
Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The...Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The solute carrier family 7(SLC7)genes are involved in activating the target of rapamycin complex 1(TORC1)signaling pathway and protein translation process,but the physiological roles of SLC7 genes in silk-producing insects have not been reported.Here,we found that amino acid signaling regulates silk protein synthesis and larval development via the L-type amino acid trans-porter 1(LAT1;also known as SLC7A5)in Bombyx mori.A total of 12 SLC7 homologs were identified in the silkworm genome,among which BmSLC7A5 was found to be a silk gland-enriched gene and may be involved in leucine transport.Bioinformatics analy-sis indicated that SLC7A5 displays high homology and a close phylogenetic relationship in silk-producing insects.Subsequently,we found that leucine treatment significantly in-creased silk protein synthesis by improving the transcription and protein levels of silk genes.Furthermore,systemic and silk gland-specific knockout of BmSLC7A5 led to de-creased silk protein synthesis by inhibiting TORC1 signaling,and somatic mutation also resulted in arrested development from the 5th instar to the early pupal stage.Altogether,our study reveals that BmSLC7A5 is involved in regulating silk protein synthesis and larval development by affecting the TORC1 signaling pathway,which provides a new strategy and target for improving silk yield.展开更多
MicroRNAs(miRNAs)are important regulators of nearly all aspects of biological processes in eukaryotes.During the biogenesis of miRNAs,the RNase III enzyme Dicer processes double-strand precursor miRNAs into mature miR...MicroRNAs(miRNAs)are important regulators of nearly all aspects of biological processes in eukaryotes.During the biogenesis of miRNAs,the RNase III enzyme Dicer processes double-strand precursor miRNAs into mature miRNAs and promotes the assembly of RNA-induced silencing complexes(RISCs).Dicer has been reported to participate in a wide range of physiological processes,including development and immunity,in some insect species.However,the physiological roles of Dicer in lepidopterans remain poorly understood.In this study,we investigated the function of Bombyx mori Dicer1.We first performed sequence alignment and found that the sequence of functional domains of Dicer1 are varied among Lepidoptera,Diptera,Coleoptera,Blattaria,and Orthoptera.Using a binary clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 genome editing approach,we showed that BmDicer1 mutants have arrested development from the 3rd instar into the 4th instar.RNA sequencing analysis indicated that the defects in BmDicer1 mutants are due to dysregulation of genes that encode proteins involved in metabolism,protein degradation,absorption,and renin–angiotensin pathways.Analysis using quantitative real-time polymerase chain reaction showed that mutation of BmDicer1 altered expression of miRNAs and their target genes.Therefore,our study demonstrates the critical roles of BmDicer1 in miRNA biogenesis and larval development in silkworm.展开更多
Although information about embryonic and larval development of garpike,Belone belone(Linnaeus,1761),is present in the published literature,the bulk of research concerns garpike from the northeastern Atlantic Ocean and...Although information about embryonic and larval development of garpike,Belone belone(Linnaeus,1761),is present in the published literature,the bulk of research concerns garpike from the northeastern Atlantic Ocean and the Baltic Sea.The present work describes the embryonic and larval development of garpike,Belone belone,from the Adriatic Sea,and methods used for incubation of fertilized eggs in aquarium conditions.Because garpike is,as suggested by some authors,divided into subspecies,we conclude that some differences in embryonic development could also be expected.In the present study,eggs were fertilized using the dry fertilization method and were incubated in a tank equipped with aeration and constant sea water flow.Salinity and content of dissolved oxygen were constant,and the temperature varied between 19.4 and 22.3°C.Eggs were spherical,measuring 3071.9±75.73μm in diameter.Yolk sacs were homogeneous and did not contain oil globules.The first larvae hatched 329 h and 47 min after fertilization.Absorption of the yolk sac occurred 17 h–48 h after hatching and the total length of newly hatched larvae was 9.78 mm.The peculiarities observed in the embryonic and early larval development are evidence of an exceptional plasticity and adaptive potential,which could be considered as helpful features in extending the natural range of occurrence of this species.展开更多
We describe organogenesis at a histological level in American shad( Alosa sapidissima) larvae from 0 until 45 days after hatching(DAH). Larval development was divided into four stages based on the feeding mode,externa...We describe organogenesis at a histological level in American shad( Alosa sapidissima) larvae from 0 until 45 days after hatching(DAH). Larval development was divided into four stages based on the feeding mode,external morphological features,and structural changes in the organs: stage 1(0–2 DAH),stage 2(3–5 DAH),stage 3(6–26 DAH) and stage 4(27–45 DAH). At early stage 2(3 DAH),American shad larvae developed the initial digestive and absorptive tissues,including the mouth and anal opening,buccopharyngeal cavity,oesophagus,incipient stomach,anterior and posterior intestine,differentiated hepatocytes,and exocrine pancreas. The digestive and absorptive capacity developed further in stages 2 to 3,at which time the pharyngeal teeth,taste buds,gut mucosa folds,differentiated stomach,and gastric glands could be observed. Four defined compartments were discernible in the heart at 4 DAH. From 3 to 13 DAH,the excretory systems started to develop,accompanied by urinary bladder opening,the appearance and development of primordial pronephros,and the proliferation and convolution of renal tubules. Primordial gills were detected at 2 DAH,the pseudobranch was visible at 6 DAH,and the filaments and lamellae proliferated rapidly during stage 3. The primordial swim bladder was first observed at 2 DAH and started to inflate at 9 DAH; from then on,it expanded constantly. The spleen was first observed at 8 DAH and the thymus was evident at 12 DAH. From stage 4 onwards,most organs essentially manifested an increase in size,number,and complexity of tissue structure.展开更多
Cathepsin L gene is a member of the cysteine proteinase gene group. In this study Cathepsin L gene was isolated from Kuruma shrimp Marsupenaeusjaponicus （Mj-Cathepsin L） and the full-length DNA sequence was 1 963 bp...Cathepsin L gene is a member of the cysteine proteinase gene group. In this study Cathepsin L gene was isolated from Kuruma shrimp Marsupenaeusjaponicus （Mj-Cathepsin L） and the full-length DNA sequence was 1 963 bp. Mj-Cathepsin L protein showed high homologies with other Cathepsin L proteins documented in vertebrates, mollusks and other crustaceans. Expression analysis of Mj-Cathepsin L gene in different tissues revealed that it was predominant in hepatopancreas. During early ontogenetic development stages Mj-Cathepsin L showed a development-regulated expression, and the Mj-Cathepsin L showed a molting stage-regulated expression during the five molting stages, inferring its role in the ontogenic development of M.japonicus. Two kinds of forms of Mj- Cathepsin L protein： pro-Cathepsin L and Cathepsin L were measured in hepatopancreas, stomach and intestine by Western Blotting.展开更多
Phospholipids are used to improve the growth and survival of Portunus trituberculatus,a widely cultured crab species in China.However,only total phospholipids or several classes are applied in crab diets.In this study...Phospholipids are used to improve the growth and survival of Portunus trituberculatus,a widely cultured crab species in China.However,only total phospholipids or several classes are applied in crab diets.In this study,we employed a targeted lipidomic method to investigate the comprehensive phospholipid composition in P.trituberculatus larvae and reveal the changing phospholipid profile over the larval development.Results showed that P.trituberculatus larvae contain 112 phospholipid species belonging to 10 phospholipid classes,in which phosphatidylcholine(PC)and phosphatidylethanolamine(PE)species are the most abundant,and PC,PE,phosphatidic acid,and phosphatidylserine(PS)are with high concentrations.The levels of all phospholipids significantly changed with larval development,which was highlighted by the downward parabolic changes in PE,phosphatidylglycerol,phosphatidylino-sitol,PS,lysophosphatidic acid,and sphingomyelin levels.In addition,nearly all phospholipid species were depleted at the M stage,which probably contributed to the mass mortality of crab larvae.These findings on the composition and alterations of phospholipids in P.trituberculatus larvae provide novel perspectives for the targeted supplementation of phospholipids in crab diets.Our work also highlights the use of targeted UHPLC-MS lipidomics in understanding the changes of phospholipids during crab development.展开更多
As an organophosphorus compound that frequently detected in water samples,triphenyl phosphate(TPhP)has been showed to have multiple toxicological effects on aquatic species.However,no attention has been paid to its po...As an organophosphorus compound that frequently detected in water samples,triphenyl phosphate(TPhP)has been showed to have multiple toxicological effects on aquatic species.However,no attention has been paid to its potential impact on non-model amphibian species.Here,tadpoles of the Zhenhai brown frog(Rana zhenhaiensis)were exposed to different concentrations of TPh P(0,0.02 and 0.1 mg/L)throughout the developmental period to assess physiological and meta bolic impacts of TPh P exposure on amphibian larvae.After 30-day TPh P exposure,the developmental stage of tadpoles from the high-concentration treatment appeared to be more advanced than that from the other two treatments,but other measured traits(including body size,tail length and liver weight)did not differ among treatments.Metabolite profiles in tadpole livers based on liquid chromatographymass spectrometry(LC-MS)revealed a distinct metabolic disorder in exposed animals.Specifically,significant changes in various hepatic amino acids(such as glutamine,glutamate,valine and leucine)were observed.Overall,our results indicated that chronic TPhP exposure potentially caused developmental and hepatic physiological changes in R.zhenhaiensis tadpoles,although its impact on tadpole growth appeared to be minor.展开更多
Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profi...Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements.Larvae were collected at the egg stage and 0,2,4,7,12,17,24 days after hatching(DAH)for analysis.Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH.Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH.Pepsin activity increased sharply beginning 17 DAH.Lipase activity increased significantly 4 DAH and increased progressively with larval growth.Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease.High free amino acid(FAA) content was detected in starry flounder eggs(110.72mg/g dry weight).Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae.Most FAAs(except lysine and methionine) decreased >50%in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae.Changes in the protein amino acid(PAA) profile were much milder than those observed for FAAs.Most PAAs increased gradually during larval development,except lysine and phenylalanine.The percentages of free threonine,valine,isoleucine,and leucine decreased until the end of the trial,whereas the protein forms of these four AAs followed the opposite trend.A comparison of the essential AA composition of live food(rotifers,Artemia nauplii,and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA.These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food.Moreover,the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed.展开更多
Assessing the determinants of reproductive success is critical but often complicated because of complex interactions between parental traits and environmental conditions occurring during several stages of a reproducti...Assessing the determinants of reproductive success is critical but often complicated because of complex interactions between parental traits and environmental conditions occurring during several stages of a reproductive event.Here,we used a simplified ecological situation-an amphibian species lacking post-oviposition parental care-and a laboratory approach to investigate the relationships between parental(both maternal and paternal)phenotypes(body size and condition)and reproductive success(fecundity,egg size,embryonic and larval duration,larval and metamorphic morphology).We found significant effects of maternal phenotype on fecundity,hatching success,and tadpole size,as well as on the duration of larval development.Interestingly,and more surprisingly,we also found a potential contribution of the paternal phenotype occurring during early(embryonic development duration)offspring development.Although our study focused on life-history traits such as body size and development duration,additional mechanisms involving physiological costs of development may well mediate the relationships between parental phenotypes and offspring development.Future studies are required to decipher the mechanisms underlying our findings in order to clarify the mechanistic basis of the links between parental phenotypes and offspring development.展开更多
文摘Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficiency is limited. For this purpose, the embryonic and larval development of reciprocal crosses of Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) were studied under laboratory conditions. The fertilization rate varied from 63.33% to 92%, while the hatching rate ranged from 55.68% to 76% with the highest value in hybrids ♀Cg × ♂Cj. Crosses between ♀Cj × ♂Cj, ♀Cg × ♂Cj and ♀Cj × ♂Cg had embryonic stages similar to those of the pure sib ♀Cg x ♂Cg. All crosses, however, had different timing for the various embryological stages. Hatching occurred at 32 h 15 min and 38 h for ♀Cj × ♂Cj and ♀Cj × ♂Cg, and 23 h and 23 h 30 min, respectively, for ♀Cg × ♂Cg and ♀Cg × ♂Cj. However, both crosses produced viable larvae until the first external feeding. The external morphological features of the larvae were completely formed by the 10th day after hatching. The body forms of the crosses at this time were indistinguishable from the pure sib. This study thus laid the groundwork for further comparative studies on hybrid performance and characterization.
基金financial support from Zhang Zidao Sland Group Co.,Ltd. for the project (99801214)
文摘Bacterial community presumably plays an essential role in inhibiting pathogen colonization and maintaining the health of scallop larvae, but limiting data are available for Yesso scallop （Patinopecten yessoensisis Jay, 1857） larval development stages. The aim of this study was to characterize and compare the bacterial communities associating with Yesso scallop larval development at fertilized egg S l, trochophora S2, D-shaped larvae S3, umbo larvae S4, and juvenile scallop S5 stages by Illumina high-throughput sequencing. Genomic DNA was extracted from the larvae and their associating baetera, and a gene segment covering V3-V4 region of 16S rRNA gene was amplified and sequenced using an Illumina Miseq sequencer. Overall, 106760 qualified sequences with an average length of 449 bp were obtained. Sequences were compared with those retrieved from 16S rRNA gene databases, and 4 phyla, 7 classes, 15 orders, 21 families, 31 genera were identified. Proteobacteria was predominant phylum, accounting for more than 99%, at all 5 larval development stages. At genus level, Pseudomonas was dominant at stages S1 （80.60%）, S2 （87.77%） and S5 （68.71%）, followed by Photobacterium （17.06%） and Aeromonas （1.64%） at stage S1, Serratia （6.94%）, Stenotrophomonas （3.08%） and Acinetobacter （1.2%） at stage S2, Shewanella （25.95%） and Pseudoalteromonas （4.57%） at stage S5. Moreover, genus Pseudoal- teromonas became dominant at stages S3 （44.85%） and S4 （56.02%）, followed by Photobacterium （29.82%）, Pseudomonas （11.86%）, Aliivibrio （8.60%） and Shewanella （3.39%） at stage S3, Pseudomonas （18.16%）, Aliivibrio （14.29%）, Shewanella （4.11%）, Psychro- monas （4.04%） and Psychrobacter （1.81%） at stage S4. From the results, we concluded that the bacterial community changed sig- nificantly at different development stages of Yesso Scallop larvae.
基金Supported by the Special Scientific Research Funds for Central Non-Profit Institutes,CAFS(No.2014A01 YYOl)the National Basic Research Program of China(973 Program)(No.2011CB409805)+1 种基金the Earmarked Fund for Modern Agro-Industry Technology Research System(No.CARS-48)the National Key Technology R&D Program of China(No.2011BAD45B01)
文摘We evaluated the effect of pH on larval development in larval Pacific oyster(Crassostrea gigas) and blood cockle(Arca inflata Reeve).The larvae were reared at pH 8.2(control),7.9,7.6,or 7.3beginning 30 min or 24 h post fertilization.Exposure to lower pH during early embryonic development inhibited larval shell formation in both species.Compared with the control,larvae took longer to reach the D-veliger stage when reared under pH 7.6 and 7.3.Exposure to lower pH immediately after fertilization resulted in significantly delayed shell formation in the Pacific oyster larvae at pH 7.3 and blood cockle larvae at pH 7.6 and 7.3.However,when exposure was delayed until 24 h post fertilization,shell formation was only inhibited in blood cockle larvae reared at pH 7.3.Thus,the early embryonic stages were more sensitive to acidified conditions.Our results suggest that ocean acidification will have an adverse effect on embryonic development in bivalves.Although the effects appear subtle,they may accumulate and lead to subsequent issues during later larval development.
基金This work was supported by the National Natural Science Foundation of China(No.41676140)the Major Agriculture Program of Ningbo(No.2017C110007)+1 种基金the Modern Technology System of Agricultural Industry(No.CARS48)the K C Wong Magana Fund in Ningbo University.
文摘The swimming crab Portunus trituberculatus is an economically important marine crustacean species.Here isobaric tags for relative and absolute quantitation(iTRAQ)analysis were used to identify proteins that are differentially expressed during larval development of P.trituberculatus to elucidate the underlying mechanisms.In comparison with the first zoea stage(Z1),3980 proteins were identified from 32789 peptides,which were matched with 115497 spectrums.A total of 241 proteins were screened with significantly differential expressions in all development stages.These 241 proteins are involved in various biological processes,such as cytoskeleton organization,protein synthesis,energy production and substance metabolism,physiological activities,and transport.Cluster analyses of the 241 differentially expressed proteins led to the generation of four protein clusters based on the overall similarity in protein expression patterns.Exactly 54,70,36,and 45 proteins clustered in profiles 10(0,0,0,−1,0,0),15(0,1,0,1,0,1),18(0,1,2,2,1,0),and 19(0,1,2,3,4,5),respectively.Muscle development and exoskeleton renewal were important processes throughout the development stages.In addition,protein synthesis,degradation,and digestion actively occurred,especially at the Z4 stage.These results provide novel insights into the mechanisms underlying larval development of swimming crab and can assist in larval rearing.
文摘With increasing demand in China for the mitten crab larvae, understanding its survival mechanism gets more important. This research focused on the effects of temperature and Ca 2+ on the larval growth and development. Eriocheir sinensis larvae were reared in laboratory under 21 different combinations of temperature (15, 20, 25℃) and Ca 2+ content (120, 130, 140, 150, 160, 170, 180 mg/l) and constant salinity (20) and pH (8). The results suggested that the survival rate increases with temperature and Ca 2+ content. These combinations of temperature and Ca 2+ content maximized survival rate in our study and it may be the optimum water environmental conditions for culturing the larvae. To predict surviving larvae number under different water environmental conditions, 21 dynamic mathematical models were developed. This for the first time observation of the zoeal Ⅵ larvae of the Changjiang River E. sinensis population showed that they occurred under stressed water environmental conditions: temperature of 15℃ and Ca 2+ content of 120,130 mg/l.
基金Thiswork was supported bygrants fromthe NationalNaturalScience FoundationofChina(31772532)the China Postdoctoral Science Foundation(2022MD713704)the Chongqing Science and Technology Bureau(cstc2021ljcyj-bshX0222 and jbky20210004).
文摘Insects produce silk to form cocoons,nests,and webs,which are important for their survival and reproduction.However,little is known about the molecular mecha-nism of silk protein synthesis at the translation level.The solute carrier family 7(SLC7)genes are involved in activating the target of rapamycin complex 1(TORC1)signaling pathway and protein translation process,but the physiological roles of SLC7 genes in silk-producing insects have not been reported.Here,we found that amino acid signaling regulates silk protein synthesis and larval development via the L-type amino acid trans-porter 1(LAT1;also known as SLC7A5)in Bombyx mori.A total of 12 SLC7 homologs were identified in the silkworm genome,among which BmSLC7A5 was found to be a silk gland-enriched gene and may be involved in leucine transport.Bioinformatics analy-sis indicated that SLC7A5 displays high homology and a close phylogenetic relationship in silk-producing insects.Subsequently,we found that leucine treatment significantly in-creased silk protein synthesis by improving the transcription and protein levels of silk genes.Furthermore,systemic and silk gland-specific knockout of BmSLC7A5 led to de-creased silk protein synthesis by inhibiting TORC1 signaling,and somatic mutation also resulted in arrested development from the 5th instar to the early pupal stage.Altogether,our study reveals that BmSLC7A5 is involved in regulating silk protein synthesis and larval development by affecting the TORC1 signaling pathway,which provides a new strategy and target for improving silk yield.
基金supported by grants from National Natural Science Foundation of China(31830093,32100381).The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘MicroRNAs(miRNAs)are important regulators of nearly all aspects of biological processes in eukaryotes.During the biogenesis of miRNAs,the RNase III enzyme Dicer processes double-strand precursor miRNAs into mature miRNAs and promotes the assembly of RNA-induced silencing complexes(RISCs).Dicer has been reported to participate in a wide range of physiological processes,including development and immunity,in some insect species.However,the physiological roles of Dicer in lepidopterans remain poorly understood.In this study,we investigated the function of Bombyx mori Dicer1.We first performed sequence alignment and found that the sequence of functional domains of Dicer1 are varied among Lepidoptera,Diptera,Coleoptera,Blattaria,and Orthoptera.Using a binary clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 genome editing approach,we showed that BmDicer1 mutants have arrested development from the 3rd instar into the 4th instar.RNA sequencing analysis indicated that the defects in BmDicer1 mutants are due to dysregulation of genes that encode proteins involved in metabolism,protein degradation,absorption,and renin–angiotensin pathways.Analysis using quantitative real-time polymerase chain reaction showed that mutation of BmDicer1 altered expression of miRNAs and their target genes.Therefore,our study demonstrates the critical roles of BmDicer1 in miRNA biogenesis and larval development in silkworm.
文摘Although information about embryonic and larval development of garpike,Belone belone(Linnaeus,1761),is present in the published literature,the bulk of research concerns garpike from the northeastern Atlantic Ocean and the Baltic Sea.The present work describes the embryonic and larval development of garpike,Belone belone,from the Adriatic Sea,and methods used for incubation of fertilized eggs in aquarium conditions.Because garpike is,as suggested by some authors,divided into subspecies,we conclude that some differences in embryonic development could also be expected.In the present study,eggs were fertilized using the dry fertilization method and were incubated in a tank equipped with aeration and constant sea water flow.Salinity and content of dissolved oxygen were constant,and the temperature varied between 19.4 and 22.3°C.Eggs were spherical,measuring 3071.9±75.73μm in diameter.Yolk sacs were homogeneous and did not contain oil globules.The first larvae hatched 329 h and 47 min after fertilization.Absorption of the yolk sac occurred 17 h–48 h after hatching and the total length of newly hatched larvae was 9.78 mm.The peculiarities observed in the embryonic and early larval development are evidence of an exceptional plasticity and adaptive potential,which could be considered as helpful features in extending the natural range of occurrence of this species.
基金Supported by the National Spark Program of China(No.2012GA690001)
文摘We describe organogenesis at a histological level in American shad( Alosa sapidissima) larvae from 0 until 45 days after hatching(DAH). Larval development was divided into four stages based on the feeding mode,external morphological features,and structural changes in the organs: stage 1(0–2 DAH),stage 2(3–5 DAH),stage 3(6–26 DAH) and stage 4(27–45 DAH). At early stage 2(3 DAH),American shad larvae developed the initial digestive and absorptive tissues,including the mouth and anal opening,buccopharyngeal cavity,oesophagus,incipient stomach,anterior and posterior intestine,differentiated hepatocytes,and exocrine pancreas. The digestive and absorptive capacity developed further in stages 2 to 3,at which time the pharyngeal teeth,taste buds,gut mucosa folds,differentiated stomach,and gastric glands could be observed. Four defined compartments were discernible in the heart at 4 DAH. From 3 to 13 DAH,the excretory systems started to develop,accompanied by urinary bladder opening,the appearance and development of primordial pronephros,and the proliferation and convolution of renal tubules. Primordial gills were detected at 2 DAH,the pseudobranch was visible at 6 DAH,and the filaments and lamellae proliferated rapidly during stage 3. The primordial swim bladder was first observed at 2 DAH and started to inflate at 9 DAH; from then on,it expanded constantly. The spleen was first observed at 8 DAH and the thymus was evident at 12 DAH. From stage 4 onwards,most organs essentially manifested an increase in size,number,and complexity of tissue structure.
基金The National High-tech R&D Program of China(863 Program)under contract No.2012AA10A409-03the Project of China Agriculture Research System under contract No.CARS-47+1 种基金the Project of Xiamen Southern Ocean Research Center under contract No.14CZY033HJ07China Spark Program under contract No.2015GA720002
文摘Cathepsin L gene is a member of the cysteine proteinase gene group. In this study Cathepsin L gene was isolated from Kuruma shrimp Marsupenaeusjaponicus （Mj-Cathepsin L） and the full-length DNA sequence was 1 963 bp. Mj-Cathepsin L protein showed high homologies with other Cathepsin L proteins documented in vertebrates, mollusks and other crustaceans. Expression analysis of Mj-Cathepsin L gene in different tissues revealed that it was predominant in hepatopancreas. During early ontogenetic development stages Mj-Cathepsin L showed a development-regulated expression, and the Mj-Cathepsin L showed a molting stage-regulated expression during the five molting stages, inferring its role in the ontogenic development of M.japonicus. Two kinds of forms of Mj- Cathepsin L protein： pro-Cathepsin L and Cathepsin L were measured in hepatopancreas, stomach and intestine by Western Blotting.
基金the National Natural Science Foundation of China(Nos.41673076,32073024)the Col-laborative Promotion Program of Zhejiang Province Ag-ricultural Technology of China(No.2020XTTGSC03)the China Agriculture Research System-CARS48 and K.C.Wong Magna Fund in Ningbo University are acknowle-dged.
文摘Phospholipids are used to improve the growth and survival of Portunus trituberculatus,a widely cultured crab species in China.However,only total phospholipids or several classes are applied in crab diets.In this study,we employed a targeted lipidomic method to investigate the comprehensive phospholipid composition in P.trituberculatus larvae and reveal the changing phospholipid profile over the larval development.Results showed that P.trituberculatus larvae contain 112 phospholipid species belonging to 10 phospholipid classes,in which phosphatidylcholine(PC)and phosphatidylethanolamine(PE)species are the most abundant,and PC,PE,phosphatidic acid,and phosphatidylserine(PS)are with high concentrations.The levels of all phospholipids significantly changed with larval development,which was highlighted by the downward parabolic changes in PE,phosphatidylglycerol,phosphatidylino-sitol,PS,lysophosphatidic acid,and sphingomyelin levels.In addition,nearly all phospholipid species were depleted at the M stage,which probably contributed to the mass mortality of crab larvae.These findings on the composition and alterations of phospholipids in P.trituberculatus larvae provide novel perspectives for the targeted supplementation of phospholipids in crab diets.Our work also highlights the use of targeted UHPLC-MS lipidomics in understanding the changes of phospholipids during crab development.
基金a grant from the Natural Science Foundation of Zhejiang Province(LY15C030006)。
文摘As an organophosphorus compound that frequently detected in water samples,triphenyl phosphate(TPhP)has been showed to have multiple toxicological effects on aquatic species.However,no attention has been paid to its potential impact on non-model amphibian species.Here,tadpoles of the Zhenhai brown frog(Rana zhenhaiensis)were exposed to different concentrations of TPh P(0,0.02 and 0.1 mg/L)throughout the developmental period to assess physiological and meta bolic impacts of TPh P exposure on amphibian larvae.After 30-day TPh P exposure,the developmental stage of tadpoles from the high-concentration treatment appeared to be more advanced than that from the other two treatments,but other measured traits(including body size,tail length and liver weight)did not differ among treatments.Metabolite profiles in tadpole livers based on liquid chromatographymass spectrometry(LC-MS)revealed a distinct metabolic disorder in exposed animals.Specifically,significant changes in various hepatic amino acids(such as glutamine,glutamate,valine and leucine)were observed.Overall,our results indicated that chronic TPhP exposure potentially caused developmental and hepatic physiological changes in R.zhenhaiensis tadpoles,although its impact on tadpole growth appeared to be minor.
基金Supported by the Scientific and Technological Development Plan Project of Yantai City(No.2013ZH348)the Aquatic Animal Nutrition and Feed Research and Innovation Demonstration Platform(No.201301001)the National Special Research Fund for Non-Profit Marine Sector(No.1205025)
文摘Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements.Larvae were collected at the egg stage and 0,2,4,7,12,17,24 days after hatching(DAH)for analysis.Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH.Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH.Pepsin activity increased sharply beginning 17 DAH.Lipase activity increased significantly 4 DAH and increased progressively with larval growth.Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease.High free amino acid(FAA) content was detected in starry flounder eggs(110.72mg/g dry weight).Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae.Most FAAs(except lysine and methionine) decreased >50%in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae.Changes in the protein amino acid(PAA) profile were much milder than those observed for FAAs.Most PAAs increased gradually during larval development,except lysine and phenylalanine.The percentages of free threonine,valine,isoleucine,and leucine decreased until the end of the trial,whereas the protein forms of these four AAs followed the opposite trend.A comparison of the essential AA composition of live food(rotifers,Artemia nauplii,and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA.These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food.Moreover,the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed.
基金Funding was provided by the CNRS,the Agence de lʹEau Loire-Bretagne,the Agence de lʹEau Adour-Garonne,the Conseil Départemental des Deux-Sèvres,the Région Nouvelle-Aquitaine(Multistress 2017-1R20110,Aquastress 2018-1R20214,Amphitox 2019-1R20216)the ANSES(BiodiTox project 2019/1/031)the Plan dʹAction National ECOPHYTO(OFB-21-0941),and the CPER Econat.
文摘Assessing the determinants of reproductive success is critical but often complicated because of complex interactions between parental traits and environmental conditions occurring during several stages of a reproductive event.Here,we used a simplified ecological situation-an amphibian species lacking post-oviposition parental care-and a laboratory approach to investigate the relationships between parental(both maternal and paternal)phenotypes(body size and condition)and reproductive success(fecundity,egg size,embryonic and larval duration,larval and metamorphic morphology).We found significant effects of maternal phenotype on fecundity,hatching success,and tadpole size,as well as on the duration of larval development.Interestingly,and more surprisingly,we also found a potential contribution of the paternal phenotype occurring during early(embryonic development duration)offspring development.Although our study focused on life-history traits such as body size and development duration,additional mechanisms involving physiological costs of development may well mediate the relationships between parental phenotypes and offspring development.Future studies are required to decipher the mechanisms underlying our findings in order to clarify the mechanistic basis of the links between parental phenotypes and offspring development.
