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The present study aimed to compare the nutritional effects of cholesterol, bile acids, and combination of cholesterol with bile acids in plant-based diets on juvenile genetically improved farmed tilapia (GIFT; Oreochromis niloticus). The isonitrogenous (321 g/kg crude protein) and isolipidic (76 g/kg crude fat) diets (Con diet) were based on plant protein sources, which included corn gluten meal, soybean meal, cottonseed meal and rapeseed meal. The Con diet was supplemented with 12 g/kg cholesterol (CHO diet), 0.2 g/kg bile acids (BAs diet), a combination of 12 g/kg cholesterol and 0.2 g/kg bile acids (CHO–BAs diet), respectively. Each diet was fed to three tanks in an indoor recirculating aquaculture system for 9 weeks. Results showed that compared to the Con group, fish had a higher weight gain rate, hepatosomatic index, and a lower feed conversion ratio in the CHO–BAs group. The highest levels of whole-fish fat and ash were found in the Con group. Serum parameters, including activities of alanine aminotransferase (ALT) and aspartate transaminase (AST), along with levels of glucose (GLU) and triglyceride (TG) except for total cholesterol (TCHO), were lower in the CHO, BAs, and CHO–BAs groups than those in the Con group (P < 0.001). Histological examination revealed that fish in the Con group exhibited severe hepatocyte vacuolization and diminished hepatocyte proliferation. Gene expression analysis indicated that the transcriptional levels of bile acid metabolism-related genes (including fxr, fgf19, bsep) were up-regulated in the CHO–BAs group (P < 0.05), whereas cholesterol metabolism-related genes (acly and hmgcr) were down-regulated in both CHO and CHO–BAs groups (P < 0.001). Moreover, UPLC-MS/MS analysis revealed that the higher taurine-conjugated bile acids (T-BAs), followed by free bile acids (Free-BAs) and glycine (G-BAs) were determined in tilapia bile. Among these, taurochenodeoxycholic bile acid was the predominant bile acid. Dietary bile acids supplementation also increased the proportion of T-BAs (tauro β-muricholic acid and taurodehydrocholic acid) while decreasing Free-BAs in the fish bile. In conclusion, the incorporation of cholesterol with bile acids into plant-based diets can effectively reduce cholesterol uptake, suppress bile acids synthesis, enhance bile acids efflux, and promote hepatocyte proliferation, which is helpful for maintaining the normal liver morphology in tilapia, and thus improving its growth performance.

Nile tilapia (Oreochromis niloticus) is a worldwide farmed fish and has been widely used for the study on comparative immunology in teleosts. It is well known that cadmium (Cd) can cause a variety of adverse effects in fish. However, data on the effects of Cd in fish liver and the defensive mechanisms of these effects using transcriptome approach are relatively scarce to date. In this study, by using an RNA sequencing approach, the gene expression profiling was performed in livers of tilapia exposed to 0 (control), 50, 100, and 200 μg/L of Cd for 2 months. The results showed that exposure to 50 μg/L Cd altered the expressions of 911 genes, while exposure to 100 and 200 μg/L Cd resulted in 4318 and 3737 differentially expressed genes compared to the control. Weighted correlation network analysis (WGCNA) and gene ontology (GO) analysis identified a 14-gene network linked to the immune system development. Further, in a fuzzy analysis, the GO term immune system development was enriched in cluster 3, and gene expression decreased with increasing Cd levels in a concentration-dependent manner. The qPCR and RNA-seq results identified 4 genes, i.e., dnmt3bb.1, sf3b1, SMARCAL1, and zap70, as convenient potential biological indicators for detecting waterborne Cd. The present results help systematically understand the effects of Cd on the hepatic transcriptome in tilapia.

BackgroundInterspecies hybridization is an important breeding method to generate fishes with heterosis in aquaculture. Using this method, hybrid Nile tilapia (Oreochromis niloticus, ♀) × blue tilapia (Oreochromis aureus, ♂) has been produced and widely farmed due to its growth and appetite superiorities. However, the genetic mechanism of these advanced traits is still not well understood. Ghrelin is a crucial gene that regulates growth and appetite in fishes. In the present study, we focused on the expression characteristics and its regulation of ghrelin in the hybrid.ResultsThe tissue distribution analysis showed that ghrelin was predominantly expressed in the stomach in the hybrid. Ghrelin was more highly expressed in the stomach in the hybrid and Nile tilapia, compared to blue tilapia, showing a nonadditive pattern. Two single-nucleotide polymorphism (SNP) sites were identified including T/C and C/G from the second exon in the ghrelin gene from Nile tilapia and blue tilapia. By pyrosequencing based on the SNP sites, the allele-specific expression (ASE) of ghrelin in the hybrid was assayed. The result indicated that ghrelin in the hybrid showed higher maternal allelic transcript ratios. Fasting significantly increased ghrelin overall expression at 4, 8, 12, 24, and 48 h. In addition, higher maternal allelic transcript ratios were not changed in the fasting hybrids at 48 h. The cis and trans effects were determined by evaluating the overall expression and ASE values in the hybrid. The expression of ghrelin was mediated by compensating cis and trans effects in hybrid.ConclusionIn summary, the present lines of evidence showed the nonadditive expression of ghrelin in the hybrid tilapia and its regulation by subgenomes, offering new insight into gene expression characteristics in hybrids.

The escalating problem of copper (Cu) and cadmium (Cd) pollution in aquatic environments poses a significant threat to the ovarian tissue and reproductive capacity of fish, hindering the development of the aquaculture industry. However, the combined effects of Cu and Cd on fish gonadal development remain unclear. In this study, the fish species Nile tilapia was stressed with rearing water containing 300 μg/L Cu2+ and 100 μg/L Cd2+ for 30 days, followed by an intraperitoneal injection of luteinizing hormone-releasing hormone (LHRH-α) and human chorionic gonadotropin (HCG) at various concentrations. We investigated the ovarian transcriptome profiles before and after injection. Prior to injection, combined treatment with Cu and Cd resulted in reproductive dysfunction and metal ion imbalance in tilapia. Transcriptomic profiling revealed differential gene annotation concentrated in the MAPK signaling pathway and regulation of GTPase activity. Post-injection, all concentrations of LHRH-α and HCG groups showed an upregulated gonadosomatic index (G.S.I) and higher levels of vitellogenin (VTG), gonadotropin-releasing hormone (GnRH), gonadotropin (GTH), and estrogen (E2) in serum compared to the negative control group. Transcriptomic analysis revealed alterations in various ovarian signaling pathways, preliminarily revealing the in vivo molecular mechanisms and differences in LHRH-α and HCG. The findings from this study could help us better understand how to counteract the effects of combined Cu and Cd exposure on tilapia ovarian development, which has significant implications for the Nile tilapia aquaculture industry.

【Objective】The study was conducted to explore the effects of compound probiotics on the growth performance and muscle quality of tilapia and aquaculture water environment under zero-water exchange condition, with a view to providing references for emission reduction as well as efficient and healthy aquaculture in fishery industry.【Method】Taking tilapia as research object and regular water exchange aquaculture mode as control group, a full-cycle aquaculture test of tilapia was carried out by adding compound probiotics under zero-water exchange condition (experimental group), and a comparative analysis on growth performance, muscle quality of tilapia and main water quality, microbial community structure was conducted.【Result】Under zero-water exchange condition, the survival rate of tilapia of the experimental group was 83.3%, significantly higher than the control group; feed coefficient was 2.3, significantly lower than the control group; as for the muscle quality, crude protein content of the experiment group was 22.17%, significantly higher than the control group. Main water quality of the experimental group stayed stable through the full-cycle of aquaculture period, the water temperature was 26.5-30.5℃, pH was 7.0-8.0, nitrite of water sample was 0.039 mg/L, significantly lower than the control group. The dominant bacteria was similar between experimental and control groups, however, the diversity index of the experimental group was higher than that of the control group.【Conclusion】Under zero-water exchange condition, adding compound probiotics in aquaculture water improved the survival rate of tilapia, reduced the feed conversion rate, and improved the muscle quality of tilapia. Meanwhile, the experimental group had strong improving effects on the water environment of aquaculture to some certain extent, and increased the diversity index of bacterial community. During the whole aquaculture production cycle, the zero-water exchange aquaculture of tilapia with high-activity compound probiotics had a good substitution effect for the conventional aquaculture model which relies on frequent water changes to release environmental pressure.

Circular RNA (circRNA) is an endogenous biomolecule in eukaryotes. It has tissue- and cell-specific expression patterns and can act as a microRNA sponge or competitive endogenous RNA. Although circRNA has been found in several species in recent years, the expression profiles in fish gonad are still not fully understood. We detected the expression of circRNA in the ovary, testis, and sex-changed gonad of tilapia by high-throughput deep sequencing, and circRNA-specific computing tools. A total of 20,607 circRNAs were obtained, of which 141 were differentially expressed in the testis and ovary. Among these circRNAs, 135 circRNAs were upregulated and 6 circRNAs were downregulated in female fish. In addition, GO annotation and KEGG pathway analysis of the host genes of circRNAs indicated that these host genes were mainly involved in adherens junction, androgen production, and reproductive development, such as ZP3, PLC, delta 4a, ARHGEF10, and HSD17b3. It is worth noting that we found that circRNAs in tilapia gonads have abundant miRNA-binding sites. Among them, 935 circRNAs have a regulatory effect on miR-212, 856 circRNAs have a regulatory effect on miR-200b-3p, and 529 circRNAs have a regulatory effect on miR-200b-5p. Thus, our findings provide a new evidence for circRNA–miRNA networks in the gonads in tilapia.

Parachromis managuensis is a native cichlid fish from Central America that has been recently introduced to Southern China. The present study aimed to identify the adaptive evolution of P. managuensis using transcriptome data and provided genetic information regarding this novel fish in China. We obtained the transcriptome sequences from a mixed cDNA library of P. managuensis. A total of 19,419,739 raw reads were obtained from the cDNA library. The de novo assembly generated a total of 102,977 unigenes. The genomic data of Atlantic cod (Gadus morhua) and Nile tilapia (Oreochromis niloticus) were used to identify orthologs by phylogenetic analysis. Based on the phylogeny, we detected 105 positively selected genes in P. managuensis from 6197 orthologous unigenes. The GO annotation revealed that the T-box protein 20 (TBX20) in the category ‘Immune system process’ was positively selected and four positively selected genes were found in the ‘Immune system’ pathway by KEGG analysis. Among them, the genes encoding for phosphoinositide 3-kinase adapter protein 1, ras-related protein Rap-1b, complement factor I and probable ATP-dependent RNA helicase DHX58 were associated with immune adaptation. These findings can provide additional information regarding the natural selection of P. managuensis and can be used as a representative example for future comparative transcriptome and immunological studies.

Estradiol is a steroid hormone that regulates multiple physiological processes in teleosts. The present study aimed to investigate the role of 17β-estradiol (E2) in lipid metabolism in the liver of Nile tilapia. Waterborne exposures were performed using different concentrations of E2 (0 ng/L, 10 ng/L, 25 ng/L and 50 ng/L) and its receptor inhibitor ICI 182,780 (ICI) (1 μM/L). Subsequently, the survival rate and hepatosomatic index (HSI) of tilapia were measured and no significant changes could be found following E2 exposure. Gene expression of VLDL assembly, lipoprotein lipase and lipoprotein receptor, fatty acid uptake and triacylglycerol metabolism were detected by qPCR. E2 exposure up-regulated the expression of lipid metabolism-related genes MTP, LPL, CD36 and DGAT2. ICI exposure inhibited the level of LDLR in the liver of tilapia. This showed that E2 stimulated the MTP expression and promoted VLDL synthesis. In addition, E2 waterborne exposure induced secretion of LDLR and LPL and promoted fatty acid production. Moreover, E2 up-regulated the expression of CD36 to accelerate the transport of fatty acids. Overall, our study provides new information for evaluating Environmental estrogens (EEs) effects on farming fishes and endocrinological regulation on lipid metabolism in fish.

Hybridization is a common breeding technique that can improve germplasm through heterosis in aquaculture. However, the regulation of key gene expression, including the details of transcriptional level changes at the beginning of hybridization events, remains largely undefined, especially in teleosts. In this study, by interspecies crossing between two pure lines of Nile tilapia and blue tilapia, we obtained a hybrid tilapia population as a model to elucidate heterosis, and we traced the molecular outcomes of growth hormone (GH) expression and allele-specific expression (ASE) in hybrids. The hybrids display growth vigor compared to their parents in the 120-day growth trial. GH mRNA expression was uniquely expressed in the pituitary. Higher GH expression was found in the hybrid than the midparent value, in both males and females, showing a nonadditive pattern. We identified four single-nucleotide polymorphism sites between Nile tilapia and blue tilapia. Subsequently, by pyrosequencing, we found asymmetric allelic expression in hybrids with higher maternal allelic transcript ratios in both males and females. Fasting significantly increased GH expression in hybrids, but asymmetric allelic expression was not affected by feeding or fasting conditions. Finally, we identified cis and trans effects via overall expression and ASE values in the hybrid, which showed that the cis and trans effects promoted the expression of maternal subgenome in the hybrid, contributing to the expression superiority of GH in hybrid tilapia. Taken together, the results of our study first illustrated the concept of GH expression superiority and its formation mechanism in hybrid fish with growth vigor.

This study investigated the molecular mechanisms involved in ovarian transcriptomic responses in Nile tilapia under different photoperiod regimes. Histological analysis indicated that ovarian development was significantly affected by photoperiod. The photoperiods tested were as follows: LD (12 h light:12 h dark), LL (24 h light:0 h dark), and DD (0 h light:24 h dark). The longer photoperiod (LL) was shown to induce ovary development earlier than LD and DD. Next, ovary transcriptome levels were sequenced and analyzed. These data indicated that 988, 992, and 1,036 differentially expressed genes (DEGs) were detected by comparing LD–LL, LD–DD, and LL–DD. A number of genes that may be involved in photoperiod-specific regulation of ovarian development were observed. These findings may be useful for investigating the molecular mechanisms underlying light-induced ovarian development in Nile tilapia.

MicroRNAs (miRNAs) are endogenous non-coding small RNAs which play important roles in the regulation of gene expression by cleaving or inhibiting the translation of target gene transcripts. Thereinto, some specific miRNAs show regulatory activities in gonad development via translational control. In order to further understand the role of miRNA-mediated posttranscriptional regulation in Nile tilapia (Oreochromis niloticus) ovary and testis, two small RNA libraries of Nile tilapia were sequenced by Solexa small RNA deep sequencing methods. A total of 9,731,431 and 8,880,497 raw reads, representing 5,407,800 and 4,396,281 unique sequences were obtained from the sexually mature ovaries and testes, respectively. After comparing the small RNA sequences with the Rfam database, 1,432,210 reads in ovaries and 984,146 reads in testes were matched to the genome sequence of Nile tilapia. Bioinformatic analysis identified 764 mature miRNA, 209 miRNA-5p and 202 miRNA-3p were found in the two libraries, of which 525 known miRNAs are both expressed in the ovary and testis of Nile tilapia. Comparison of expression profiles of the testis, miR-727, miR-129 and miR-29 families were highly expressed in tilapia ovary. Additionally, miR-132, miR-212, miR-33a and miR-135b families, showed significant higher expression in testis compared with that in ovary. Furthermore, the expression patterns of the miRNAs were analyzed in different developmental stages of gonad. The result showed different expression patterns were observed during development of testis and ovary. In addition, the identification and characterization of differentially expressed miRNAs in the ovaries and testis of Nile tilapia provides important information on the role of miRNA in the regulation of the ovarian and testicular development and function. This data will be helpful to facilitate studies on the regulation of miRNAs during teleosts reproduction.

[Obbectives] This study aimed to determine the infection pathway and taget organs of Streptocoocus agalactia! in GIFT strain of Nile tilapia, thus providing theoretical basis for the breeding of disease-resistant tilapia and development of S. agalactiae vaccines. [Methods] GIFT strain of Nile tilapia was inoculated by S. agalactia! through intraperitoneal injection, oral gavage and in vitro immersion. The giil, spleen, liver and small intestine tissues of infected tilapia were collected for pathomorphological observation. Immunohistochemical localization was conducted using rabbit anti-S. agalactiae serum to identify the distribution pattern of S. agalactiae in various tiisues of tilapia and its taget organs via diiferent infection pathways. [Results] GIFT strain of Nile tilapia could be infected by S. agalactiae via three artificial inoculation modes. SpecificHy, pathological changes occurred at 2 h post-inoculation in intraperitoneal injection and oral gavage groups, whereas tilapia in in vitro immersion group showed pathological changes at 5 h post-inoculation, and the lesion intensity in in vitro immersion group was slighter than that in intraperitoneal injection and oral gavage groups. Immunohistochemical localization indicated that the appearance time of positive signals in intraperitoneal injection group demonstrated an order of spleen--diver and giil--•smaH intestine; positive signals in oral gavage group appeared in the order of smaH intestine--»gill and spleen--diver; the appearance time of positive signals in in vitro immersion group showed an order of giil--• spleen--• liver and smaH intestine. [Conclusions] GIFT strain of Nile tilapia could be infected by S. agalactiae via intraperitoneal injection, oral gavage and in vitro immersion. The corresponding positive signas for pathogen infection were preferentiaHy present in the spleen, intestine and giil tiisues. Thus, preventing S. agalactiae contamination in aquaculture water and food sources is an efective measure to control the outbreak of S. agalactiae infections in tilapia under natural aquaculture conditions. [ABSTRACT FROM AUTHOR]

Cichlid fishes have played an important role in evolutionary biology and aquaculture industry. Nile tilapia ( Oreochromis niloticus), blue tilapia ( Oreochromis aureus) and Mozambique tilapia ( Oreochromis mossambicus), the useful models in studying evolutionary biology within Cichlid fishes, are also mainly cultured species in aquaculture with great economic importance. In this paper, the complete nucleotide sequence of the mitochondrial genome for O. niloticus and O. aureus were determined and phylogenetic analyses from mitochondrial protein-coding genes were conducted to explore their phylogenetic relationship within Cichlids. The mitogenome is 16,625 bp for O. niloticus and 16,628 bp for O. aureus, containing the same gene order and an identical number of genes or regions with the other Cichlid fishes, including 13 protein-coding genes, two rRNA genes, 22 tRNA genes and one putative control region. Phylogenetic analyses using three different computational algorithms (maximum parsimony, maximum likelihood and Bayesian method) show O. niloticus and O. mossambicus are closely related, and O. aureus has remotely phylogenetic relationship from above two fishes. [ABSTRACT FROM AUTHOR]

Background: The Chinese mitten crab, Eriocheir sinensis, is one of the most studied and economically important crustaceans in China. Its transition from a swimming to a crawling method of movement during early development, anadromous migration during growth, and catadromous migration during breeding have been attractive features for research. However, knowledge of the underlying molecular mechanisms that regulate these processes is still very limited. Findings: A total of 258.8 gigabases (Gb) of raw reads from whole-genome sequencing of the crab were generated by the Illumina HiSeq2000 platform. The final genome assembly (1.12 Gb), about 67.5 % of the estimated genome size (1.66 Gb), is composed of 17,553 scaffolds (>2 kb) with an N50 of 224 kb. We identified 14,436 genes using AUGUSTUS, of which 7,549 were shown to have significant supporting evidence using the GLEAN pipeline. This gene number is much greater than that of the horseshoe crab, and the annotation completeness, as evaluated by CEGMA, reached 66.9 %. Conclusions: We report the first genome sequencing, assembly, and annotation of the Chinese mitten crab. The assembled draft genome will provide a valuable resource for the study of essential developmental processes and genetic determination of important traits of the Chinese mitten crab, and also for investigating crustacean evolution. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae is an important pathogenic bacterium causing great economic loss in Nile tilapia (Oreochromis niloticus) culture. Resistant and susceptible groups sharing the same genome showed significantly different resistance to S. agalactiae in the genetically improved farmed tilapia strain of Nile tilapia. The resistance mechanism is unclear. We determined genome-wide DNA methylation profiles in spleen of resistant and susceptible O. niloticus at 5 h postinfection with S. agalactiae using whole-genome bisulfite sequencing. The methylation status was higher in the spleen samples from resistant fish than in the susceptible group. A total of 10,177 differentially methylated regions were identified in the two groups, including 3725 differentially methylated genes (DMGs) (3129 hyper-DMGs and 596 hypo-DMGs). The RNA sequencing showed 2374 differentially expressed genes (DEGs), including 1483 upregulated and 891 downregulated. Integrated analysis showed 337 overlapping DEGs and DMGs and 82 overlapping DEGs and differentially methylated region promoters. By integrating promoter DNA methylation with gene expression, we revealed four immune-related genes (Arnt2, Nhr38, Pcdh10, and Ccdc158) as key factors in epigenetic mechanisms contributing to pathogen resistance. Our study provided systematic methylome maps to explore the epigenetic mechanism and reveal the methylation loci of pathogen resistance and identified methylation-regulated genes that are potentially involved in defense against pathogens. [ABSTRACT FROM AUTHOR]