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Sodium dichloroisocyanurate (NaDCC, C3Cl2N3NaO3) is a solid chlorine-containing product that is widely used as a disinfectant in living environments, which has potential toxic effects on human and rats. Phascolosoma esculenta is a species native to the southeast coast of China and can be used as an indicator organism. In the present study, 150 P. esculenta were used to determine the LC50 of NaDCC for P. esculenta, then 100 P. esculenta were used to analysis the change of histopathology, oxidative stress and transcriptome after NaDCC exposure. The results showed that the LC50 of NaDCC for 48 h was 50 mg/L. NaDCC stress induced pathological events in P. esculenta, including blisters, intestinal structural damage and epithelial cell ruptured or even loss. The highest and lowest intestinal activity of superoxide dismutase in individual survivors was detected at 12 h and 72 h, respectively. Malondialdehyde levels in the intestine declined gradually from 3 h and increased at 9 h, and peaked at 12 h. Total antioxidant capacity declined at 3 h and dropped below the levels of control group after 9 h. Transcriptome sequencing analysis yielded a total of 48.65 Gb of clean data. A total of 34,759 new genes were found including 957 differentially expressed genes (DEGs). The DEGs were significantly enriched in ferroptosis, response to chemicals, response to stress, immune system, ion transport, cell death, oxidation-reduction, cellular homeostasis, protein ubiquitination, and protein neddylation. Additionally, the levels of detoxification enzymes, such as glutathione-S-transferase, cytochrome P450, ABC, UDP-glycosyltransferase and SLC transporters of endogenous and exogenous solutes were significantly changed. Overall, the results provide reference for reasonable use of disinfectants during farming, and also provide insight into the mechanisms related to NaDCC toxicity in P. esculenta.
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Desinfectantes , Triazinas , Humanos , Animales , Ratas , Desinfectantes/toxicidad , Desinfectantes/química , Intestinos , Estrés Oxidativo , Perfilación de la Expresión GénicaRESUMEN
To understand the environmental adaptations among sessile bivalves lacking adaptive immunity, a series of analyses were conducted, with special emphasis on the widely distributed C. ariakensis. Employing Pacbio sequencing and Hi-C technologies, whole genome for each of a C. ariakensis (southern China) and C. hongkongensis individual was generated, with the contig N50 reaching 6.2 and 13.0 Mb, respectively. Each genome harbored over 30,000 protein-coding genes, with approximately half of each genome consisting of repeats. Genome alignment suggested possible introgression between C. gigas and C. ariakensis (northern China), and re-sequencing data corroborated this result and indicated significant gene flow between C. gigas and C. ariakensis. These introgressed candidates, well-represented by genes related to immunity and osmotic pressure, may be associated with environmental stresses. Gene family dynamics modeling suggested immune-related genes were well represented among the expanded genes in C. ariakensis. These outcomes could be attributed to the spread of C. ariakensis.
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Crassostrea , Animales , Crassostrea/genética , Secuenciación Completa del Genoma , ChinaRESUMEN
Heavy metal accumulation in freshwater ecosystem has become one of the major aquatic environmental concerns for freshwater flora and fauna due to their higher stability and bioaccumulation as well as bio-magnification properties. Furthermore, passing through the food web, these heavy metals affect human populations ultimately. This study assessed the heavy metal accumulation in Cirrhinus mrigala in spring, autumn, and winter at different locations (I, II, and III) of Panjnad headwork. Furthermore, the human health risk assessment for the consumption of C. mrigala from the sampling locations was also carried out. Fish were collected from upper (I), middle (II), and lower (III) stream of Panjnad on a monthly basis. The current study evaluated the accumulation of Aluminum (Al), Arsenic (As), Barium (Ba), and Lead (Pb) in various fish organs (liver, kidney, gills, fins, skin, muscles and bones) and assessed their potential hazard to human health through health risk assessment indicators. The results demonstrated a significant difference (p < 0.05) in heavy metal accumulation in different fish organs, seasons, and locations. The accumulation of Al, As, Ba, and Pb were considerably higher in liver and kidney as compared to the other body organs and followed a trend of liver > kidney > gills > fins > skin > bones > muscle and the overall mean concentrations of metals in different body tissues of C. mrigala were in the order of Al > As > Ba > Pb. The results also concluded that C. mrigala caught from the Panjnad headwork is not safe for human consumption due to higher values of TTHQIng (3.76), THQIng for Ba (3.27) and CRIng for As (6.4742).
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(1) Background: Singapore grouper iridovirus (SGIV) can cause extensive fish deaths. Therefore, developing treatments to combat virulent SGIV is of great economic importance to address this challenge to the grouper aquaculture industry. Green tea is an important medicinal and edible plant throughout the world. In this study, we evaluated the use of green tea components against SGIV infection. (2) Methods: The safe working concentrations of green tea components were identified by cell viability detection and light microscopy. Additionally, the antiviral activity of each green tea component against SGIV infection was determined with light microscopy, an aptamer (Q5c)-based fluorescent molecular probe, and reverse transcription quantitative PCR. (3) Results: The safe working concentrations of green tea components were green tea aqueous extract (GTAE) ≤ 100 µg/mL, green tea polyphenols (TP) ≤ 10 µg/mL, epigallocatechin-3-gallate (EGCG) ≤ 12 µg/mL, (-)-epigallocatechin (EGC) ≤ 10 µg/mL, (-)-epicatechin gallate (EGC) ≤ 5 µg/mL, and (-)-epicatechin (EC) ≤ 50 µg/mL. The relative antiviral activities of the green tea components determined in terms of MCP gene expression were TP > EGCG > GTAE > ECG > EGC > EC, with inhibition rates of 99.34%, 98.31%, 98.23%, 88.62%, 73.80%, and 44.31%, respectively. The antiviral effect of aptamer-Q5c was consistent with the results of qPCR. Also, TP had an excellent antiviral effect in vitro, wherein the mortality of fish in only the SGIV-injection group and TP + SGIV-injection group were 100% and 11.67%, respectively. (4) Conclusions: In conclusion, our results suggest that green tea components have effective antiviral properties against SGIV and may be candidate agents for the effective treatment and control of SGIV infections in grouper aquaculture.
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Lubina , Infecciones por Virus ADN , Enfermedades de los Peces , Iridovirus , Ranavirus , Animales , Antivirales/farmacología , Antivirales/uso terapéutico , Infecciones por Virus ADN/veterinaria , Iridovirus/genética , Ranavirus/fisiología , TéRESUMEN
Aquaculture offers a promising source of economic and healthy protein for human consumption, which can improve wellbeing. Viral diseases are the most serious type of diseases affecting aquatic animals and a major obstacle to the development of the aquaculture industry. In the background of antibiotic-free farming, the development and application of antibiotic alternatives has become one of the most important issues in aquaculture. In recent years, many medicinal plants and their active pharmaceutical ingredients have been found to be effective in the treatment and prevention of viral diseases in aquatic animals. Compared with chemical drugs and antibiotics, medicinal plants have fewer side-effects, produce little drug resistance, and exhibit low toxicity to the water environment. Most medicinal plants can effectively improve the growth performance of aquatic animals; thus, they are becoming increasingly valued and widely used in aquaculture. The present review summarizes the promising antiviral activities of medicinal plants and their active pharmaceutical ingredients against aquatic viruses. Furthermore, it also explains their possible mechanisms of action and possible implications in the prevention or treatment of viral diseases in aquaculture. This article could lay the foundation for the future development of harmless drugs for the prevention and control of viral disease outbreaks in aquaculture.
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Plantas Medicinales , Virosis , Virus , Animales , Antibacterianos/uso terapéutico , Acuicultura , Preparaciones Farmacéuticas , Virosis/tratamiento farmacológico , Virosis/prevención & control , Virosis/veterinariaRESUMEN
The genus Dendronereis Peters, 1854 is characterized in the polychaete family Nereididae by its feather-shaped branchiae on the anterior segments. In this study, we present the first complete mitogenome of Dendronereis, represented by D. chipolini Hsueh, 2019, collected from Beibu Gulf, China. The nucleotide composition is biased toward A + T nucleotides, accounting 31.5% for A, 22.3% for C, 14.7% for G and 31.5% for T. The assembled mitogenome is 15,763 bp in length, with a typical set of 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA), 2 ribosomal RNA (rRNA), and 1 non-coding control region. All genes are encoded on H-strand. The control region is 1260 bp in length and located between tRNA-Gly and tRNA-Met. Phylogenetic study showed that D. chipolini is arranged with high support into the clade of Namanereidinae. The complete mitogenome provides important molecular data for investigating the phylogeny and evolution of the nereid animals.
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It is difficult to distinguish the sexes of Trachinotus ovatus based on appearance, and little data about sex-determining genes are available for this species. Here, we generated 200 F2 individuals using the parents R404 and R403. DNA samples were collected from 50 individuals of each sex and aggregated into sex-specific DNA pools. Specific-locus amplified fragment sequencing was integrated with bulked segregant analysis to detect candidate sex-associated genes. Approximately 3,153,153 high-quality single-nucleotide polymorphism (SNP) markers and 135,363 high-quality insertion-deletion (Indel) markers were generated. Six candidate regions within chromosome 14, encompassing 132 candidate genes, were identified as closely related to sex. Based on annotations, six genes (EVM0019817, EVM0004192, EVM0001445, EVM0005260, EVM0014734, and EVM0009626) were predicted to be closely associated with sex. These results present an efficient genetic mapping approach that lays a foundation for molecular sex discrimination in T. ovatus.
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Proteínas de Peces/genética , Peces/genética , Polimorfismo de Nucleótido Simple , Análisis para Determinación del Sexo/métodos , Animales , Mapeo Cromosómico/métodos , Femenino , Mutación INDEL , Masculino , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Scavenger receptor class B (SRB) is a multifunctional protein in animals that participates in physiological processes, including recognition of a wide range of ligands. Astaxanthin is a major carotenoid found in shrimp. However, the molecular mechanism of astaxanthin and SRB protein binding has not been reported. RESULTS: In the present study, a member of the SRB subfamily, named PmSRB, was identified from the transcriptome of black tiger shrimp (Penaeus monodon). The open reading frame of PmSRB was 1557 bp in length and encoded 518 amino acids. The structure of PmSRB included a putative transmembrane structure at the N-terminal region and a CD36 domain. Multiple sequence alignment indicated that the CD36 domain were conserved. Phylogenetic analysis showed four separate branches (SRA, SRB, SRC, and croquemort) in the phylogenetic tree and that PmSRB was clustered with SRB of Eriocheir sinensis. Quantitative real-time polymerase chain reaction showed that the PmSRB gene was widely expressed in all tissues tested, with the highest expression level observed in the lymphoid organ and brain. Subcellular localization analysis revealed that PmSRB-GFP (green fluorescent protein) fusion proteins were predominantly localized in the cell membrane. The recombinant proteins of PmSRB showed binding activities against astaxanthin in vitro. CONCLUSIONS: PmSRB was identified and characterized in this study. It is firstly reported that PmSRB may take as an important mediator of astaxanthin uptake in shrimp.
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Animales , Penaeidae , Receptores Depuradores/metabolismo , Técnicas In Vitro , Western Blotting , Cromatografía Líquida de Alta Presión , Alineación de Secuencia , Xantófilas , Receptores Depuradores/aislamiento & purificación , Receptores Depuradores/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , TranscriptomaRESUMEN
Nanoparticulate and ionic Zn have potential impacts on the detoxification systems of organisms, and Gst genes play key roles in the detoxification of xenobiotics. In this study, we cloned the ChGstα and ChGstκ genes of C. hongkongensis, and studied their expression in gills under nanoparticulate and ionic Zn stress. The results showed that the coding sequences of the ChGstα and ChGstκ genes were 684 and 675 bp, respectively, and had no signal peptide; ChGstα was cytoplasmic, while ChGstκ was mitochondrial. The two genes were expressed in all 8 tested samples, with the most abundant expression observed in hemocytes for ChGstα and digestive glands for ChGstκ. After ZnCl2 or ZnoNP challenge, the expression of ChGstα decreased significantly in the ZnCl2 groups, and its expression was higher in the ZnoNP groups than in the ZnCl2 groups. The expression of ChGstκ was significantly decreased in the ZnCl2 and ZnoNP groups, and its expression was higher in the ZnoNP groups than in the ZnCl2 groups except at 3 h post metal Zn stress, which suggested that ChGstα and ChGstκ were more sensitive to ZnoNP than ZnCl2.
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Crassostrea/genética , Branquias/metabolismo , Glutatión Transferasa/metabolismo , Nanopartículas del Metal/toxicidad , Contaminantes Químicos del Agua/toxicidad , Zinc/toxicidad , Animales , Xenobióticos/toxicidadRESUMEN
Microplastic pollution in marine environments and organisms has received a great deal of international attention. However, the long-term field studies of microplastics are rare. Here, we assessed annual variation in microplastic abundance in the Maowei Sea, a classic mariculture bay in southern China, and analyzed the long-term accumulation in oyster tissues. U-shaped time trends of microplastics in water were observed from January to December in 2018 in the estuarine region, inner bay, and mouth bay sites, representing an inverse relationship with the local rainfall patterns. The common microplastic particles in Maowei Sea are PET/PE fibers, and polystyrene foams, which are mainly related to textile pollution and fishery activities. After one year of continuous monitoring, we did not find accumulation of microplastics in the whole soft tissues of oyster after 10% KOH digestion. No significant correlation of microplastic abundances between water and oysters was observed. The microplastic abundance in oyster was correlated with some environmental variables (i.e. salinity, pH, nutrients and total organic carbon) of the surrounding water following Spearman correlation analysis. The microplastic levels in oysters could probably be influenced by the environmental variables.
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Cherax quadricarinatus is seriously affected by multiple types of pathogens, including bacteria and viruses, and has been widely transplanted around the world. Heat shock proteins (Hsps) are a group of molecular chaperones that play important roles in promoting the proper refolding and blocking the aggregation of denatured proteins. In this study, CqHsp60, CqHsp70 and CqHsp90 from C. quadricarinatus were cloned, and their expression patterns were analysed. The CDS (coding sequence) lengths of the CqHsp60, CqHsp70 and CqHsp90 genes were 1731 bp, 1932 bp and 2199 bp, encoding 576, 643 and 732 amino acids, respectively. CqHsp60 was 99.13%, 98.78% and 88.63% identical to the corresponding sequences of Cherax cainii, Cherax destructor and Eriocheir sinensis, respectively. CqHsp70 showed 99.84%, 92.73% and 91.58% identity to the corresponding sequences of C. cainii, C. destructor and E. sinensis, while CqHsp90 was 98.25%, 98.51% and 91.41% identical with those of C. cainii, C. destructor and E. sinensis, respectively. The expression patterns of the three CqHsps were different between males and females. CqHsp60 and CqHsp70 exhibited the highest expression in the hepatopancreas of males and the gonads of females, and CqHsp90 presented the highest expression in the gonads of males and hepatopancreas of females. After pathogenic inoculation, the death trend of C. quadricarinatus at different time points was the same in association with different pathogens, with most deaths occurring within 6 h post-inoculation. The trend of CqHsp transcription at different time points was the same among the groups treated with Vibrio alginolyticus, Vibrio parahemolyticus and Aeromonas hydrophila, exhibiting upregulation first and then downregulation. The expression of CqHsp60 and CqHsp70 in the gills of living C. quadricarinatus was less than 3.5 times that in the PBS group, but in the gills of dead C. quadricarinatus under A. hydrophila inoculation, its expression was more than 5-9 times that in the PBS group. CqHsp90 expression changed dramatically in the V. alginolyticus, V. parahemolyticus and A. hydrophila groups, in which it exceeded 50 times the level in the PBS group. These results indicated that CqHsps could induce the activation of the immune system within a short time and that CqHsp90 could be used as a more effective molecular biomarker than CqHsp70 and CqHsp60 in a pathogenic bacterium-polluted environment.
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A novel Streptomyces strain (SSL-25T) was isolated from mangrove soil sampled at QinzhouBay, PR China. The isolate was observed to be Gram-stain-positive and to form greyish-white aerial mycelia that differentiated into straight spore chains with smooth-surfaced spores on International Streptomyces Project 2 medium. The cell-wall peptidoglycan was determined to contain ll-diaminopimelicacid. The cell-wall sugars were glucose and mannose. The predominant menaquinones were MK-9 (H6), MK-9 (H8) and MK-9 (H4). The major polar lipids contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and several unidentified phospholipids. The predominant cellular fatty acids were C16:0, iso-C16:0 and summed feature 3 (C16:1ω7c/C16:1ω6c). The genome size of strain SSL-25T was 8.1 Mbp with a G+C content of 71.5 mol%. Phylogenetic analysis indicated that strain SSL-25T is closely related to Streptomyces tsukubensis NRRL 18488T (99.4 % sequence similarity). However, the digital DNA-DNA hybridization (39.8 %) and average nucleotide identity (91.3 %) values between them showed that it represents a distinct species. Furthermore, the results of morphological, physiological and biochemical tests allowed further phenotypic differentiation of strain SSL-25T from S. tsukubensis NRRL 18488T. Therefore, based on these results, it is concluded that strain SSL-25T represents a novel Streptomyces species, for which the name Streptomyces qinzhouensis sp. nov. is proposed. The type strain is SSL-25T (=CICC 11054T=JCM33585T).
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Filogenia , Microbiología del Suelo , Streptomyces/clasificación , Técnicas de Tipificación Bacteriana , Pared Celular/química , China , ADN Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Grasos/química , Tamaño del Genoma , Hibridación de Ácido Nucleico , Peptidoglicano/química , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Suelo , Streptomyces/aislamiento & purificación , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMEN
Xyrias revulsus is one of the species in the China-Vietnam Collective Fishery Zone, which has a few relevant studies. In this study, the mitochondrial genome of X. revulsus was determined for the first time using next-generation sequencing; the overall base components of mitogenome consisting of 17,784 bp was 32.45% for A, 25.76% for T, 15.72% for G, 26.08% for C, and its GC content was 41.8%. The mitochondrial circular genome was composed of 13 protein-coding genes, 22 transfer RNAs, 2D-loop, and 2 ribosomal RNAs. Polygenetic analysis showed that the X. revulsus was very close to Ophisurus macrorhynchos. It can provide data reference for the analysis of genetic evolution of this species.
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An outbreak of suspected iridovirus disease in cultured hybrid grouper (âTiger Grouper Epinephelus fuscoguttatus × â Giant Grouper Epinephelus lanceolatus) occurred in the Guangxi Province in July, 2018. In this study, grouper iridovirus Guangxi (SGIV-Gx) was isolated from diseased hybrid grouper that were collected from Guangxi. Cytopathic effects were observed and identified in grouper spleen cells that were incubated with diseased tissue homogenates after 24 h, and the effects increased at 48 h postinfection. The transmission electron microscopy results showed that viral particles that were about 200 nm in diameter with hexagonal profiles were present in the cell cytoplasm of suspected virus-infected cells. The presence of SGIV-Gx (accession number: MK107821) was identified by polymerase chain reaction (PCR) and amplicon sequencing, which showed that this strain was most closely related to Singapore grouper iridovirus (AY521625.1). The detection of SGIV-Gx infection was further supported by novel aptamer (Q2c)-based detection technology. The effects of temperature and pH on viral infectivity were analyzed by using reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and cell culture. The results indicated that SGIV-Gx was resistant to exposure to pH levels 5, 7, and 7.5 for 1 h, but its infectivity was remarkably lower at pH levels 3 and 10 after 1 h. The analyses showed that SGIV-Gx was stable for 1 h at 4°C and 25°C but was inactivated after 1 h at 40, 50, and 60°C.
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Lubina , Infecciones por Virus ADN/veterinaria , Enfermedades de los Peces/virología , Ranavirus/aislamiento & purificación , Animales , China , Infecciones por Virus ADN/patología , Infecciones por Virus ADN/virología , Enfermedades de los Peces/patología , Microscopía Electrónica de Transmisión/veterinaria , Ranavirus/clasificación , Bazo/patología , Bazo/ultraestructura , Bazo/virologíaRESUMEN
Lysozyme is an important defense molecule of the innate immune system and possess high antimicrobial activities. In this study, a full-length c-type lysozyme cDNA (Fplysc) was cloned and characterized from Fenneropenaeus penicillatus. The cDNA contains an open reading frame of 477 bp encoding 158 amino acids, with 53-94% identity with those of other crustaceans. The recombinant Fplysc had antibacterial activities against Gram-positive bacteria (Streptococcus agalactiae and Micrococcus luteus) and Gram-negative bacteria (Vibrio alginolyticus and Escherichia coli), and showed antiviral activity against WSSV and IHHNV. The qRT-PCR analysis showed that Fplysc expression levels were most abundant in hemocytes and less in eyestalk. The expression levels of Fplysc were significantly upregulated in gill, intestine and hemocytes when challenged with WSSV and V. alginolyticus. Fplysc-silencling suppressed Fplysc expression in cephalothoraxes and increased mortality caused by WSSV and V. alginolyticus, and exogenous rFplysc led to a significant decrease of shrimp mortality by injecting rFplysc into Fplysc silenced shrimp, suggesting Fplysc is the important molecule in shrimp antimicrobial and antiviral response. In conclusion, the results provide some insights into the function of Fplysc in shrimp against bacterial and viral infection.
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Proteínas de Artrópodos/inmunología , Penaeidae/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/genética , Clonación Molecular , Densovirinae/fisiología , Escherichia coli/fisiología , Hemocitos , Inmunidad Innata , Micrococcus luteus/fisiología , Muramidasa/química , Muramidasa/genética , Muramidasa/metabolismo , Penaeidae/genética , Penaeidae/microbiología , Penaeidae/virología , Streptococcus agalactiae/fisiología , Vibrio alginolyticus/fisiología , Virus del Síndrome de la Mancha Blanca 1/fisiologíaRESUMEN
Cerithidea sinensis is a common and important component of mangrove ecosystem. In this study, the mitochondrial genome of C. sinensis was determined for the first time using next-generation sequencing; the overall base components of mitogenome consisting of 15633 bp was 31.14% for A, 35.70% for T, 16.65% for G, 16.51% for C, and its GC content was 33.16%. The mitochondrial circular genome was composed of 13 protein-coding genes, 22 tranfer RNAs, and 2 ribosomal RNAs. Polygenetic analysis showed that the C. sinensis was more closed to Semisulcospira libertina than Turritella bacillum and Tylomelania sarasinorum. We may speculate that the C. sinensis is evolved from freshwater species.
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Trachypenaeus curvirostris is a common and important shrimp species in the shallow waters of Indo-West Pacific. In this study, the mitochondrial genome of T. curvirostris was determined for the first time using next-generation sequencing; the overall base components of mitogenome consisting of 15968 bp was 35.16% (5614 bp) for A, 33.51% (5351 bp) for T, 11.54% (1842 bp) for G, 19.80% (3161 bp) for C, and its GC content was 31.34%. The mitochondrial circular genome was composed of 13 protein-coding genes, 22 transfer RNAs, 1 D-loop and 2 ribosomal RNAs. Polygenetic analysis showed that the T. curvirostris was more closed to Parapenaeopsis hardwickii.
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Neritina violacea is a common and important component of mangrove ecosystem. In this study, the mitogenome of N. violacea was determined for the first time using next-generation sequencing; the overall base components of mitogenome consisting of 15,710 bp was 31.37% for A, 34.91% for T, 19.47% for G, 14.25% for C, and its GC content was 33.72%. The mitogenome was composed of 13 protein-coding genes, 22 tranfer RNAs, and 2 ribosomal RNAs. Polygenetic analysis showed that the N. violacea was more close to N. Usnea and Theodoxus fluviatilis. We speculated that the N. violacea was evolved from freshwater species.
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Myostatin (MSTN), also called growth and differentiation factor-8 (GDF-8), is a member of the transforming growth factor-ß (TGF-ß) superfamily and an inhibitor of muscle differentiation and growth. In this report, we identified and characterized a MSTN gene (CnMSTN) from the scallop Chlamys nobilis. The open reading frame of CnMSTN was 1374bp in length, encoding 457 amino acids. The structure of CnMSTN included a putative signal peptide, a TGF-ß propeptide domain, and a conserved TGF-ß domain. Phylogenetic analysis showed that the CnMSTN gene was clustered in the same subgroup with the MSTN gene found in Mollusca. Quantitative real-time PCR showed that the CnMSTN gene was widely expressed in all tissues tested, with the highest expression level observed in the adductor muscle. Six single nucleotide polymorphisms (SNPs) were identified in the promoter region, but no SNP was detected in the exon regions. Association analysis showed that SNP g.-579A/C had significant effects on body mass, soft-tissue mass, and adductor muscle mass. The CC and AC genotypes of g.-579A/C had significantly higher growth trait values than that of genotype AA (P<0.05). These results suggest that CnMSTN could be used as a candidate gene for the selective breeding of C. nobilis.
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Miostatina/genética , Pectinidae/crecimiento & desarrollo , Pectinidae/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Regulación del Desarrollo de la Expresión Génica , Estudios de Asociación Genética , Miostatina/química , Pectinidae/metabolismo , Filogenia , Polimorfismo de Nucleótido Simple , ARN Mensajero/genética , ARN Mensajero/metabolismo , Homología de Secuencia de Aminoácido , Distribución TisularRESUMEN
Tumor necrosis factor receptor-associated factor 6 (TRAF6) is an important cytoplasm signal adaptor that mediates signals activated by tumor necrosis factor receptor (TNFR) superfamily and the Interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily. In the study, the full-length cDNA of a TRAF6 homolog (FpTRAF6) was identified from Fenneropenaeus penicillatus. The full-length cDNA of FpTRAF6 is 2033 bp long, with an open reading frame (ORF) encoding a putative protein of 594 amino acids, including a RING type Zinc finger, two TRAF-type Zinc fingers, and a conserved C-terminal meprin and TRAF homology (MATH) domain. The overall amino acid sequence identity between FpTRAF6 and other TRAF6s ranged from 62.7 to 94.1% for crustaceans and from 45.6 to 59.3% for mollusca. Real-time qRT-PCR indicated that FpTRAF6 was constitutively expressed in various tissues of F. penicillatus. The temporal expression patterns of FpTRAF6 mRNA were different in the different tissues after microbial challenge. FpTRAF6 was downregulated in the heart, no obvious changes in the gill, intestine and hemocytes, and upregulated in other tested tissues after WSSV challenge. After V. alginolyticus injection, FpTRAF6 was downregulated in the heart and intestine, upregulated in the gill, lymphoid organ and hematopoietic organ, and no obvious changes in other tested tissues. RNAi assay was carried out to investigate the function of FpTRAF6. The results showed that silencing FpTRAF6 gene could inhibit peroxinectin expression in vivo, and enhance the sensitivity of shrimps to WSSV and V. alginolyticus challenge, suggesting FpTRAF6 could play a positive role against bacterial and viral pathogens. In conclusion, the results of the study provide some insights into the function of FpTRAF6 in activating TLRs signaling pathway and the host defense against invading pathogens.