Genes In Common Carp Research Articles

Effect of aflatoxin B1 and sterigmatocystin on DNA repair genes in common carp

The present study aimed to investigate the short-time (24 h) effect of aflatoxin B1 (AFB1) and sterigmatocystin (STC) on the expression of hsp70, p53, gadd45, and ogg1 genes in common carp hepatopancreas. Our results showed that aflatoxin B1 and sterigmatocystin can stimulate the expression of DNA repair genes, mainly by hour 24. This significant finding contributes to our understanding of the short-term effects of these mycotoxins on ogg1 genes in common carp hepatopancreas. One-year-old common carp juveniles were randomly distributed into five groups (Control, AFB1 0.4 mg kg−1 feed, STC1 1 mg kg−1 feed, STC2 2 mg kg−1 feed, and STC3 3 mg kg−1 feed). Hepatopancreas samples were collected three times (8, 16, and 24 h) in each group. No significant ogg1 and p53 expression changes were observed at 8 and 16 h after exposure. All measured genes were upregulated by the 24th hour in aflatoxin and STC3 groups. An increase in hsp70 gene expression was detected in all groups and all sampling. A significant decrease in gadd45aa gene expression was observed in the aflatoxin B1 group at hour 8. At hour 16, there was no significant change, while at hour 24, all treated groups were significantly different from the control. In summary, our results suggest that aflatoxin B1 and sterigmatocystin can stimulate the expression of DNA repair genes, mainly by hour 24. Further investigations are needed to get information about DNA damage parallel to the DNA repair mechanisms.

Effect of dietary nutmeg (Myristica fragrans) on growth performance, antioxidant status, immune response, and gene expression of common carp (Cyprinus carpio)

The aim of this study was to assess the effects of dietary nutmeg (Myristica fragrans (MF)) on growth, innate immunity parameters of serum and mucus, antioxidant defence, and expression of immune genes in common carp (Cyprinus carpio). Three hundred fish (3.79 ± 0.02 g) in four groups were distributed in twelve glass aquariums with a capacity of 150-L. The groups were T1 or control (0% MF), T2 (0.5% MF), T3 (1% MF), and T4 (2% MF), and the fish were sampled after 42 days. According to the results, diet supplementation showed significantly higher weight gain (WG) and lower food conversion ratio (FCR). Final weight (FW) increased in groups fed 1% and 2% MF and specific growth rate (SGR) improved in those provided 1% MF (P < 0.05). Alternative complement activity (ACH50) and the total immunoglobulin (total Ig) level were found to be significantly higher in T4 and T3 groups, respectively than in the T1 (P < 0.05). However, lysozyme (LYZ) and total protein (TP) in all treatment groups showed no significant variation compared with the T1. Similarly, feeding fish with supplemented diets did not affect the antioxidant status and mucus immune parameters. Additionally, fish fed 1% MF showed a significantly higher intestinal catalase (CAT) and superoxide dismutase (SOD) gene expression (P < 0.05). However, LYZ expression significantly decreased in the fish fed MF (P < 0.05). The results of the present study revealed the addition of MF to the diet affects the expression of both antioxidant and immune genes in common carp.

New reports on iron related proteins: Molecular characterization of two ferroportin genes in common carp (Cyprinus carpio L.) and its expression pattern

Iron uptake, transport, and storage require the involvement of several proteins, including ferroportin (fpn), the sole known iron efflux transporter. Due to its critical function fpn has been studied, particularly in humans. Here, we characterized the ferroportin gene in common carp (Cyprinus carpio L.) and performed RNA-seq analysis to evaluate its constitutive transcription levels across different tissues. Our results indicate that C. carpio possesses two functional fpns with distinct expression patterns, highlighting the potential for functional divergence and expression differentiation among fpns in this species.

Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio).

Highly unsaturated fatty acids (HUFAs) are essential for mammalian health, development and growth. However, most mammals, including humans, are incapable of synthesizing n-6 and n-3 HUFAs. Fish can convert C18 unsaturated fatty acids into n-6 and n-3 HUFAs via fatty acid desaturase (Fads), in which Fads2 is a key enzyme in HUFA biosynthesis. The allo-tetraploid common carp theoretically encode two duplicated fads2 genes. The expression patterns and desaturase functions of these two homologous genes are still unknown. In this study, the full length of the fads2a and fads2b were identified in common carp (Cyprinus carpio). Expression analyses indicate that both genes were mainly expressed in the liver and the expression of fads2b is higher than fads2a at different developmental stages in carp embryos. Heterogenous expression and 3D docking analyses suggested that Fads2b demonstrated stronger ∆6 and ∆5 desaturase activities than Fads2a. The core promotor regions of fads2a and fads2b were characterized and found to have different potential transcriptional binding sites. These results revealed the same desaturase functions, but different activities of two homologues of fasd2 genes in common carp. The data showed that fads2b played a more important role in HUFA synthesis through both expression and functional analyses.

The Promoter SNPs Were Associated with Both the Contents of Poly-Unsaturated Fatty Acids (PUFAs) and the Expressions of PUFA-Related Genes in Common Carp.

The allo-tetraploid common carp encodes two duplicated fads2 genes (fads2a and fads2b) and two duplicated elovl5 genes (elovl5a and elovl5b). The coding SNPs (cSNPs) of these genes were reported to be significantly associated with the PUFA contents. Whether the promoter SNPs (pSNPs) were associated with the PUFA contents has not been reported yet. In this study, after sequencing the promoters of these four genes, we identified six pSNPs associated with the contents of PUFAs in common carp, including one elovl5a pSNP, one elovl5b pSNP, and four fads2b pSNPs. The pSNPs were predicted in the locations of transcriptional factor binding sites. Together with previously identified cSNPs in fads2b and elovl5b, the pSNPs and cSNPs of these two genes had the joint effects on the PUFA contents with higher explained percentage of phenotypic variation of the PUFA contents than single gene. The expression levels of both fads2a and fads2b were significantly positively correlated with the contents of six PUFAs. The fads2b pSNPs corresponding to higher fads2b expression levels were associated with higher PUFA contents. The pSNPs and cSNPs will be useful for the future selection breeding of common carp with higher PUFA contents.

Disruption of T-box transcription factor eomesa results in abnormal development of median fins in Oujiang color common carp Cyprinus carpio.

Median fins are thought to be ancestors of paired fins which in turn give rise to limbs in tetrapods. However, the developmental mechanisms of median fins remain largely unknown. Nonsense mutation of the T-box transcription factor eomesa in zebrafish results in a phenotype without dorsal fin. Compared to zebrafish, the common carp undergo an additional round of whole genome duplication, acquiring an extra copy of protein-coding genes. To verify the function of eomesa genes in common carp, we established a biallelic gene editing technology in this tetraploidy fish through simultaneous disruption of two homologous genes, eomesa1 and eomesa2. We targeted four sites located upstream or within the sequences encoding the T-box domain. Sanger sequencing data indicated the average knockout efficiency was around 40% at T1-T3 sites and 10% at T4 site in embryos at 24 hours post fertilization. The individual editing efficiency was high to about 80% at T1-T3 sites and low to 13.3% at T4 site in larvae at 7 days post fertilization. Among 145 mosaic F0 examined at four months old, three individuals (Mutant 1-3) showed varying degrees of maldevelopment in the dorsal fin and loss of anal fin. Genotyping showed the genomes of all three mutants were disrupted at T3 sites. The null mutation rates on the eomesa1 and eomesa2 loci were 0% and 60% in Mutant 1, 66.7% and 100% in Mutant 2, and 90% and 77.8% in Mutant 3, respectively. In conclusion, we demonstrated a role of eomesa in the formation and development of median fins in Oujiang color common carp and established an method that simultaneously disrupt two homologous genes with one gRNA, which would be useful in genome editing in other polyploidy fishes.

Dominant Elongase Activity of Elovl5a but Higher Expression of Elovl5b in Common Carp (Cyprinus carpio).

Most diploid freshwater and marine fish encode one elovl5 elongase, having substrate specificity and activities towards C18, C20 and C22 polyunsaturated fatty acids (PUFAs). The allo-tetraploid common carp is hypothesized to encode two duplicated elovl5 genes. How these two elovl5 genes adapt to coordinate the PUFA biosynthesis through elongase function and expression divergence requires elucidation. In this study, we obtained the full-length cDNA sequences of two elovl5 genes in common carp, named as elovl5a and elovl5b. Functional characterization showed that both enzymes had elongase activity towards C18, C20 and C22 PUFAs. Especially, the activities of these two enzymes towards C22 PUFAs ranged from 3.87% to 8.24%, higher than those in most freshwater and marine fish. The Elovl5a had higher elongase activities than Elovl5b towards seven substrates. The spatial-temporal expression showed that both genes co-transcribed in all tissues and development stages. However, the expression levels of elovl5b were significantly higher than those of elovl5a in all examined conditions, suggesting that elovl5b would be the dominantly expressed gene. These two genes had different potential transcriptional binding sites. These results revealed the complicated roles of elovl5 on PUFA synthesis in common carp. The data also increased the knowledge of co-ordination between two homoeologs of the polyploid fish through function and expression divergence.

Interleukin (IL)-22 in common carp (Cyprinus carpio L.): Immune modulation, antibacterial defense, and activation of the JAK-STAT signaling pathway

Interleukin (IL)-22 is an IL-10 family cytokine secreted by CD4+ T cells and plays an important role in regulating inflammation and infection elimination. IL-22 homologues have been reported in the teleost, but the functions of IL-22 are still unclear. In this study, we identified two duplicated IL-22 genes in common carp (Cyprinus carpio L.), termed Cc_IL-22A and Cc_IL-22B. Sequence analysis showed that Cc_IL-22A and Cc_IL-22B had four conserved cysteine residues, which could form two intra-chain disulfide bridges. The Cc_IL-22A and Cc_IL-22B were constitutively expressed in various tissues, with the highest expression in the gill. The mRNA expression levels of Cc_IL-22A and Cc_IL-22B were significantly up-regulated in the gill, intestine, head kidney, and spleen of common carp challenged with Aeromonas. hydrophila. In vivo study showed that the expression levels of pro-inflammatory cytokines were significantly up-regulated in the head kidney and spleen when Cc_IL-22A or Cc_IL-22B were over-expressed. Furthermore, the over-expression of Cc_IL-22A and Cc_IL-22B indicated a protective effect on tissues, with only lymphocytic infiltration observed in comparison to the control and pcN3 groups, without obvious change in tissue morphology. Similar stimulatory effects of rIL-22A and rIL-22B were observed in vitro. When HKLs were stimulated with rIL-22A or rIL-22B, the expression levels of critical signaling molecules in the Janus kinase (JAK) and signal transducer and activator of transcription (STAT) pathway were significantly induced, including JAK1, JAK3, STAT1, and STAT3, as well as pro-inflammatory cytokines (IL-1β and TNF-α). Together, these results suggest that Cc_IL-22A and Cc_IL-22B may regulate inflammatory responses through the JAK-STAT signaling pathway and have a significant impact on the immune defense of common carp against bacterial infection. Therefore, our study provides a new perspective on the functions of Cc_IL-22A and Cc_IL-22B in the immune defense mechanism of fish.

Adiponectin and adiponectin receptors in common carp (Cyprinus carpio): Tissue distribution and their expressions in response to high-carbohydrate and high-lipid diets

In order to investigate the roles of adiponectin in glucose and lipid metabolism in common carp, the tissue distribution of adiponectin and its receptor genes in common carp were firstly detected in this study, and then the effects of high-carbohydrate (45%) and high-lipid (11%) diets on their expressions were studied in the feeding trial. The results showed that adipoqa and adipoqb mRNA levels were highest in the red muscle, followed by the heart and white muscle. Adiponectin receptor genes were widely expressed in all tested tissues. Two subtypes of adiponectin receptor 1 genes (adipor1a and adipor1b) were expressed at the highest level in the brain, while adipor2 mRNA was highly expressed in the heart and red muscle. The high-carbohydrate diet significantly up-regulated adipoqa and adipoqb mRNA level in the heart of common carp, while the high-lipid diet significantly promoted adipoqa mRNA expression in the red muscle and heart, compared to the control diet. For adiponectin receptor gene expression, a high-carbohydrate diet up-regulated adipor2 mRNA level in the hepatopancreas, while a high-lipid diet significantly promoted adipor1a, adipor1b and adipor2 mRNA expression in the red muscle and hepatopancreas. The results showed that both high-carbohydrate and high-lipid diets could induce adiponectin genes expression in common carp, and AdipoRs expression was more likely to be increased by the high-lipid diet than the high-carbohydrate diet. This study suggested that adiponectin and its receptors were involved in the metabolic regulation of fish under high dietary carbohydrate and lipid levels.

Molecular characterizations, immune modulation, and antibacterial activity of interleukin-17A/F1a and interleukin-17A/F1b in common carp Cyprinus carpio

Interleukin-17 (IL-17), as a pro-inflammatory cytokine family, mediates different pro-inflammatory mediators in various cell types (e.g., epithelial cells, macrophages, endothelial cells, and fibroblasts), which play an important role in defending against pathogens. The IL-17A/F1 genes have recently been reported in fish. However, the functions of these genes are still unclear. In this study, we identified two duplicated IL-17A/F1 genes in common carp (Cyprinus carpio L.), namely, CcIL-17A/F1a and CcIL-17A/F1b. Sequence analysis showed that CcIL-17A/F1a and CcIL-17A/F1b proteins had four conserved cysteine residues, which could form two intra-chain disulfide bridges. Homology comparison displayed that the deduced amino acid sequences of CcIL-17A/F1a and CcIL-17A/F1b shared 31.1%–77.3% and 32.5%–75.7% of sequence similarity to IL-17A/F1 homologues from other fish species, respectively. The mRNA expression levels of CcIL-17A/F1a and CcIL-17A/F1b were obviously increased in gill and head–kidney of fish challenged with A. hydrophila. The recombinant protein rCcIL-17A/F1a and rCcIL-17A/F1b could enhance the expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IFN-γ, and TNF-α) and chemokines (CXCL8 and CXCL20). The 3 × Flag eukaryotic expression vectors to express protein rCcIL-17A/F1a (or rCcIL-17A/F1b) were constructed and intramuscularly injected in common carp. The rCcIL-17A/F1a (or rCcIL-17A/F1b) could be successfully expressed in vivo. Four immune-related genes, namely, CD4, CD8, TNF-α, and IgM, were also significantly induced to be expressed at higher mRNA levels compared with the control. The pretreatment with CcIL-17A/F1a or CcIL-17A/F1b could markedly increase the survival rate of common carp challenged with A. hydrophila. Our results demonstrated that CcIL-17A/F1a or CcIL-17A/F1b plays an important role in immune responses and immune defense against bacteria. CcIL-17A/F1a or CcIL-17A/F1b could also be potentially used as an immunopotentiator to prevent diseases in common carp.

Comparative Transcriptome Analysis of Organ-Specific Adaptive Responses to Hypoxia Provides Insights to Human Diseases

The common carp is a hypoxia-tolerant fish, and the understanding of its ability to live in low-oxygen environments has been applied to human health issues such as cancer and neuron degeneration. Here, we investigated differential gene expression changes during hypoxia in five common carp organs including the brain, the gill, the head kidney, the liver, and the intestine. Based on RNA sequencing, gene expression changes under hypoxic conditions were detected in over 1800 genes in common carp. The analysis of these genes further revealed that all five organs had high expression-specific properties. According to the results of the GO and KEGG, the pathways involved in the adaptation to hypoxia provided information on responses specific to each organ in low oxygen, such as glucose metabolism and energy usage, cholesterol synthesis, cell cycle, circadian rhythm, and dopamine activation. DisGeNET analysis showed that some human diseases such as cancer, diabetes, epilepsy, metabolism diseases, and social ability disorders were related to hypoxia-regulated genes. Our results suggested that common carp undergo various gene regulations in different organs under hypoxic conditions, and integrative bioinformatics may provide some potential targets for advancing disease research.

Effect of secondary attack by Aeromonas hydrophila on the expression level of hif genes in common carp (Cyprinus carpio).

Hypoxia-inducible factors (hifs) are involved in infectious diseases inflammatory reactions, and immune regulation. Common carp, a representative allotetraploid species that has undergone genome-wide replication events, has important research value. In this study, common carp were infected twice with Aeromonas hydrophila. Liver tissues of common carp were collected at 4h, 12h, 24h, 48h, 3days, 7days post-first infection and 4h, 12h, 24h post-second infection. The mRNA levels of hif genes were determined at different time points. The hif2a-2, hif3a-2, hif3b-1 and hif3b-2 expression levels in the infected group were upregulated when compared with those in the control group, whereas the expression levels of other genes were downregulated after the second infection. This indicates that the effect of A.hydrophila infection on gene expression pattern is dependent on the host, pathogen, infected tissue and gene. Pressure analysis of the hif gene family revealed that the non-synonymous substitution to synonymous substitution ratio of 12 hif genes was <1, which indicated that they were in a state of purification and selection. Combined with the differences between copy genes, the polyclonal antibodies against Hif1b-1 and Hif1b-2 were successfully prepared in this study. Western blot analysis showed that the protein expression of Hif1b-1 and Hif1b-2 reached to the highest level 48h after the first infection. After the second A.hydrophila infection, the protein expression levels of Hif1b-1 and Hif1b-2 reached the highest levels at 4 and 48h, respectively. This may indicate that the Hif1b-1 and Hif1b-2 genes in common carp play an important role in the immune mechanism at the protein level. The findings of this study will lay the foundation for future studies on the immune regulatory function of common carp hif genes, which may aid in devising novel therapeutic strategies for common carp diseases, such as A.hydrophila infection.

Lipotoxic Effects of Gossypol Acetate Supplementation on Hepatopancreas Fat Accumulation and Fatty Acid Profile in Cyprinus carpio

Cottonseed meal is an excellent protein source with low price and abundant nutrients, but the existence of free gossypol restricts its wide application in animal feeds. To explore the toxic effects of free gossypol, the effects of gossypol acetate on the growth performance, hepatopancreatic fat accumulation, and muscle fatty acid composition of the common carp (Cyprinus carpio) were investigated in the present study. Carp were randomly divided into four groups, which were fed a diet supplemented with 0, 170, 340, and 680 mg/kg gossypol acetate, respectively. After 60 days of feeding, the gossypol residue in the tissue, growth performance, blood biochemical indices, hepatopancreas histology, fatty acid components, and expression of key genes involved in lipid metabolism pathways were measured. It was found that the concentrations of free gossypol were increased significantly in the serum, bile, and hepatopancreas of carp fed 340 mg/kg and 680 mg/kg gossypol acetate ( P &lt; 0.05 ). Dietary gossypol acetate had a limited effect on growth performance, and high concentrations of gossypol acetate induced excessive hepatopancreatic fat deposition and hepatocyte injury in C. carpio. In addition, gossypol acetate decreased total polyunsaturated fatty acids (PUFAs) and docosahexaenoic acid in muscle. In the hepatopancreas, decreased HSL, PPARα, and CD36 may result in fat accumulation. In the muscle, decreased FADS2 may be the key reason for the decrease in PUFAs. Taken together, these findings indicated that dietary gossypol acetate can cause lipid metabolism disorders, such as hepatopancreatic damage and significant changes in tissue fatty acid profile, which are regulated by the key lipid metabolic genes in common carp.

Conservation of members of the free fatty acid receptor gene family in common carp

Accumulating evidence supports the crucial role intestinal microbiota and their metabolites play in the homeostasis of organisms. An important class of metabolites that have been shown to affect the immune system are short chain fatty acids (SCFAs). These SCFAs can affect the host cells via passive diffusion or via ligation to receptors, among others G-protein coupled receptor (GPR) 41 and 43. GPR41 and GPR43 are both part of a family of GPR40-related receptors. Mammalian studies have shown an important role for GPR41 and GPR43 in the modulation of immune responses by SCFAs. However, up till date, no validated coding sequences for orthologues of these SCFA receptors have been published for teleost fish. We used genomic resources and cDNA cloning, to identify and validate ten coding sequences for gpr40L genes in common carp. Phylogenetic analysis showed a division into three subclasses, putatively named class a, b and c, and showed the common carp genes had a closer phylogenetic relationship to mammalian GPR43 than to mammalian GPR41. Synteny analysis revealed a clear conservation of syntenic relationships between gpr40L in the genomes of spotted gar and common carp with the relevant region in the human genome. This conservation of synteny validates the genes identified, as gpr40L. Finally, presence of gpr40L genes was investigated in silico for genomes of 25 different, mostly teleost, fish species largely confirming the observations for gpr40L of common carp with regards to both, subdivision in three subclasses a-c and conservation of synteny. Our data provide an important first step towards an understanding of the role and function of receptors for SCFAs and immunomodulation in fish.

Genome-wide identification and characterization of olfactory receptor genes in common carp (Cyprinus carpio).

The environment contains a large extent of chemical information, which could be detected as olfactory sense. Olfactory in vertebrates plays important roles on many aspects during life time, including localizing prey or food, avoiding predators, mating behavior and social communication. Considering the essential role of olfactory receptors in the specific recognition of diverse stimuli, understanding the evolutionary dynamics of olfactory receptors in teleost means a lot, especially in the allotetraploid common carp, who has undergone the fourth whole-genome duplication event. Here, we identified the whole set of olfactory receptor genes in representative teleosts and found a significant contraction in common carp when compared with other teleosts. Odorant receptor genes (OR) occupy the most among four groups of olfactory receptors, including 33 functional genes and 16 pseudogenes. Furthermore, 6 trace amine-associated receptor (TAAR) genes (including 1 pseudogene), 7 odorant-related-A receptor genes, and 10 olfactory C family receptor genes (including 3 pseudogenes) were identified in common carp. Phylogenetic and motif analysis were performed to illustrate the phylogenetic relationship and structural conservation of teleost olfactory receptors. Selection pressure analysis suggested that olfactory receptor groups in common carp were all under relaxed purifying-selection. Additionally, gene expression divergences for olfactory receptor genes were investigated during embryonic development stages of common carp. We aim to determine the abundance of common carp olfactory receptor genes, explore the evolutionary fate and expression dynamics, and provide some genomic clues for the evolution of polyploid olfactory after whole-genome duplication and for future studies of teleost olfactory.

The regulatory roles of yucca extract on the growth rate, hepato-renal function, histopathological alterations, and immune-related genes in common carp exposed with acute ammonia stress

The accumulation of ammonia in fish ponds regularly occurs in intensive aquaculture systems, and the inclusion of yucca extract is recognized as a practical solution to adsorb the waterborne ammonia. In this context, the study was planned to investigate the possible regulatory roles of yucca extract on the performances of common carp exposed to acute ammonia stress. Fish with similar initial weight were assigned to four groups (triplicates) where fish in the control group reared without ammonia exposure and without yucca treatment whereas the second group was exposed to ammonia (10 mg/L) without yucca treatment. The third group treated with yucca (0.75 mg/L) without ammonia while the fourth group exposed with ammonia and treated with yucca. The groups were named as the control, the ammonia, the yucca, and the ammonia/yucca. After 30 days, the growth performance, survival rate, total protein, albumin, and globulin of fish treated with yucca extract had the highest values (P < 0.05) followed by control and those exposed to acute ammonia stress and treated with yucca. The feed conversion ratio (FCR) displayed the lowest value in fish treated with yucca without ammonia stress while the highest FCR was in fish exposed to ammonia without yucca (P < 0.05). The uric acid and urea levels displayed the lowest value in fish treated with yucca without ammonia stress while the highest uric acid and urea were in fish exposed to ammonia without yucca (P < 0.05). The levels of blood creatinine, aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) implemented the highest values in fish exposed to acute ammonia stress without yucca while the lowest values were in the group of fish treated with yucca without ammonia exposure (P < 0.05). The expression of hepatic catalase (CAT), superoxide dismutase (SOD), and interleukin (IL-10) genes were upregulated in fish treated with yucca and downregulated in fish exposed to acute ammonia stress. Heat shock protein (HSP70), interleukin (IL-8), tumor necrosis factor-alpha (TNF-α), interferon gamma (IFN-γ), and interleukin (IL-1β) were upregulated in fish exposed to acute ammonia stress and downregulated in the group of fish treated with yucca extract (P < 0.05). The histopathological study revealed that fish exposed to acute ammonia stress had inflammatory and abnormal features while yucca extract induced anti-inflammation influences. Hence, the study concluded that the treatment of yucca extract is recommended to protect common carp from the toxicity of waterborne ammonia.

Growth Performance, Chemical Composition of Fillets, Liver and Intestinal Histology, and Expression of Lipid-Dependent Genes in Common Carp (Cyprinus carpio) Fed Artificial Diets

The study is a preface to redesign farming of carp, and the aim was to determine the influence of three artificial feeds on carp growth performance, chemical composition, intestinal and liver histology, and the expression of two main genes of fatty acid catabolism: acyl-coenzyme A oxidase 1 (acox1) and peroxisome proliferator-activated receptor  (ppar). The 61-days trials were run in set of 9 floating cages in triplicate (n=3 per diet). The results showed that zootechnical parameters (i.e. feed conversion ratio, protein efficiency ratio) significantly improved with increased level of fat in diet, for instance, the lowest FCR was noted for fish fed diet A (9.900.23% of fat). Additionally, gene expression analysis revealed that activity of acox1 and ppar in the liver depend on the fat level in carp diet. The expression of ppar and its target gene acox1 in the intestine of fish showed a distinct pattern in relation to liver samples, since feeds with high and low levels of fat exerted a comparable effect on the mRNA level of the studied genes. In conclusion, this study showed that the level of fat in common carp diets correlated with the level of lipids in the meat.

Phylogeny of Slc15 family and response to Aeromonas hydrophila infection following Lactococcus lactis dietary supplementation in Cyprinus carpio

Solute carrier 15 family (Slc15) are membrane proteins that utilize the proton gradient and negative membrane protential for the transmembrane transporter of di-/tripeptide and peptide-mimetic molecules, in addition, they also play important roles in immunoreaction. In this study, 10 Slc15 genes were identified in the common carp genome database. Comparative genomics analysis showed considerable expansion of the Slc15 genes and verified the four-round whole genome duplication (WGD) event in common carp. Phylogenetic analysis revealed all Slc15 genes of common carp were clustered into orthologous groups indicating the highly conservative during evolution. Besides, the tissues and temporal expression examined by RT-PCR and qRT-PCR showed that most of the Slc15 genes had a narrow tissue distribution and exhibited tissue-specific expression patterns. Expression divergences were observed between these copies proving function divergence after the WGD. Then, we investigated the dietary supplementation effects of three Lactococcus lactis strains on the expression of Slc15 genes in common carp infected by A. hydrophila to find an effective way to treat aquatic diseases. Almost all of the Slc15 genes had an increased expression trend in the early post-challenge stage, and reached the highest expression level at 12h post-challenge. Then, the expression level showed a bluff descent at the last two stages and the expression level reached the lowest at 48 h post-challenge. Slc15 genes expression is actively up-regulated when stimulated by inflammatory factors, which can “amplify” immune signals, and improve the body's defense against foreign invasion in the early stage of the inflammatory response. So activation of the Slc15 genes may be an effective way for infectious disease treatment. As expected, three strains improved the expression of Slc15 genes variously compared with the control/infection groups. The strain 3 of L. lactis had a better induction of Slc15 genes compared with strain 1 and strain 2. It might be applied as a potential activation of Slc15 genes for disease treatment and adding befitting L. lactis may be a good way to protect aquatilia from bacillosis.

Short-term effects of deoxynivalenol, T-2 toxin, fumonisin B1 or ochratoxin on lipid peroxidation and glutathione redox system and its regulatory genes in common carp (Cyprinus carpio L.) liver

The effects of a single oral dose of 1.82 mg kg−1 bw of T-2 and HT-2 toxin (T-2), 1.75 mg kg−1 bw deoxynivalenol (DON) and 15-acetyl DON, 1.96 mg kg−1 bw fumonisin B1 (FB1) or 1.85 mg kg−1 bw ochratoxin A (OTA) were investigated in common carp juveniles on lipid peroxidation, the parameters of the glutathione redox system including the expression of their encoding genes in a short-term (24 h) experiment. Markers of the initiation phase of lipid peroxidation, conjugated dienes, and trienes, were slightly affected by DON and OTA treatment at 16-h sampling. The termination marker, malondialdehyde, concentration increased only as an effect of FB1. Glutathione content and glutathione peroxidase activity showed significantly higher levels in the T-2 and FB1 groups at 8 h, and in the DON and FB1 groups at 16 h. The expression of glutathione peroxidase genes (gpx4a, gpx4b) showed a dual response. Downregulation of gpxa was observed at 8 h, as the effect of DON, FB1, and OTA, but an upregulation in the T-2 group. At 16 h gpx4a upregulated as an effect of DON, T-2, and FB1, and at 24 h in the DON and T-2 groups. Expression of gpx4b downregulated at 8 h, except in the T-2 group, and upregulation observed as an effect of T-2 at 24 h. The lack of an increase in the expression of nrf2, except as the effect of DON at 8 h, and a decrease in the keap1 expression suggests that the antioxidant defence system was activated at gene and protein levels through Keap1–Nrf2 independent pathways.

Combined effects of aflatoxin B1 and deoxynivalenol on the expression of glutathione redox system regulatory genes in common carp.

The purpose of the present study was to evaluate the short-term effects of aflatoxin B1 (AFB1 ) and deoxynivalenol (DON) exposure on the expression of the genes encoding the glutathione redox system glutathione peroxidase 4a (gpx4a), glutathione peroxidase 4b (gpx4b), glutathione synthetase (gss) and glutathione reductase (gsr) and the oxidative stress response-related transcription factors Kelch-like ECH-associated protein 1 (keap1) and nuclear factor-erythroid 2 p45-related factor 2 (nrf2) in liver, kidney and spleen of common carp. During the 24-hr long experiment, three different doses (5µg AFB1 and 110µg DON; 7.5µg AFB1 and 165µg DON or 10µg AFB1 and 220µg DON/kg bw) were used. The results indicated that the co-exposure of AFB1 and DON initiated free radical formation in liver, kidney and spleen, which was suggested by the increase in Nrf2 dependent genes, namely gpx4a, gpx4b, gss and gsr. Expression of keap1 gene showed upregulation after 8hr of mycotoxin exposure, and also upregulation of nrf2 gene was found in kidney after 8hr of exposure, while in the liver, only slight differences were observed. The changes in the expression of the analysed genes suggest that level of reactive oxygen species reached a critical level where other signalling pathway was activated as described by the hierarchical model of oxidative stress.