Animal Biotechnology最新文献

Copy number variations (CNVs) have become widely acknowledged as a significant source of genomic variability and phenotypic variance. To understand the genetic variants in horses, CNVs from six Indian horse breeds, namely, Manipuri, Zanskari, Bhutia, Spiti, Kathiawari and Marwari were discovered using Axiom^™ Equine Genotyping Array. These breeds differed in agro-climatic adaptation with distinct phenotypic characters. A total of 2668 autosomal CNVs and 381 CNV regions (CNVRs) were identified with PennCNV tool. DeepCNV was employed to re-validate to get 883 autosomal CNVs, of which 9.06% were singleton type. A total of 180 CNVRs were identified after DeepCNV filtering with the estimated length of 3.12 Kb-4.90 Mb. The functional analysis showed the majority of the CNVRs genes enriched for sensory perception and olfactory receptor activity. An Equine CNVs database, EqCNVdb (http://backlin.cabgrid.res.in/eqcnvdb/) was developed which catalogues detailed information on the horse CNVs, CNVRs and gene content within CNVRs. Also, three random CNVRs were validated with real-time polymerase chain reaction. These findings will aid in the understanding the horse genome and serve as a preliminary foundation for future CNV association research with commercially significant equine traits. The identification of CNVs and CNVRs would lead to better insights into genetic basis of important traits.

Copy number variations (CNV) are important genetic variations. The endogenous factors cobalamin receptor (CUBN) and MIA SH3 domain ER-derived factor 3 (MIA3) are associated with bone/muscle development and intramuscular fat deposition. There have been no reports on the effects of CUBN and MIA3 CNVs on growth traits of Chinese cattle. This study aimed to determine the correlation between the CUBN and MIA3 CNVs and growth traits in Chinese cattle. qRT-PCR was used to detect the distribution of CUBN and MIA3 CNV and the expression levels of their mRNA, and correlation analysis was conducted between CNV and growth traits. The CUBN was differentially expressed in different breeds of cattle, and CUBN CNV correlated significantly with body height, hip height, body slanting length, and hip width of Grassland Red cattle (CYH); eye muscle area of Yanbian cattle (YB) and Yan Yellow cattle (YH). MIA3 showed no CNV in CYH and YB cattle, and only one deletion type occurred in YH cattle. CUBN and MIA3 mRNA have different expression patterns in different cattle breeds and tissues. In conclusion, CUBN CNV is correlated significantly with growth traits in Chinese cattle and is a novel molecular marker that could be exploited in cattle breeding.

Myostatin (MSTN) protein, lncRNAs, and circRNAs regulate skeletal muscle growth and development. This work aims to compare the expression patterns of circRNAs and lncRNAs in the gluteus maximus tissue of wild-type (WT) and MSTN gene knockout (KO) rabbits. Within the gluteus maximus tissue of three WT and four MSTN KO rabbits, we analyzed the expression profiles of circRNAs and lncRNAs. After identifying the differently expressed RNAs, the biological pathways implicated were ascertained by performing enrichment analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). We identified differences in the expression of 251 circRNAs (79 upregulated and 172 downregulated), 176 lncRNAs (53 upregulated and 123 downregulated), and 1178 mRNAs (408 upregulated and 770 downregulated) between WT and MSTN KO rabbits. Target genes were significantly enriched in pathways associated with protein synthesis and catabolism, such as oxidative phosphorylation, ubiquitin-mediated proteolysis, the FoxO signaling pathway, and the pentose phosphate pathway, as identified through GO and KEGG enrichment analyses. The constructed network indicates that a class of circRNAs and lncRNAs is engaged in MSTN-mediated regulation of skeletal muscle development. These findings provide valuable insights for innovative therapeutic, diagnostic, and preventive approaches to muscle disorders.

Variation in litter size (LS) in sheep is linked to genetic factors, including the Zona pellucida-3 (ZP3) gene, which plays a role in ovine reproductive processes. This study examined the association between ZP3 gene variations and LS in Kari sheep. Two groups of 160 Kari ewes were analysed: one consistently producing singletons and another producing twins, with occasional triplets. Additionally, Madakhlasht sheep, which sometimes produce twins, and Balkhi sheep, which produce only singletons, were used as references. The entire ZP3 gene was amplified using PCR and sequenced at 30× with Next Generation Sequencing. Bioinformatics analysis identified 70 variants across the three breeds, located in upstream regions, introns, and exons. Notably, two point mutations and a six-nucleotide insertion were found upstream of the initiation codon in twin-producing Kari ewes, potentially affecting ZP3 expression and LS. Two missense mutations (I101L in exon 1 and R408H in exon 8) were heterozygous in twin-producing Kari ewes but homozygous in other groups, correlating with LS. Protein modelling suggested that the I101L mutation alters the binding site, potentially impacting protein function. These findings indicate that ZP3 gene variations influence reproductive efficiency and LS in sheep, with specific variants serving as potential markers for selective breeding to enhance LS.

This study aims to establish an immortalized fibroblast cell line from Mongolian sheep. Primary Mongolian sheep fibroblasts (SSF) were isolated using tissue explant and enzymatic digestion methods, followed by microscopic observation, growth curve plotting, and karyotype analysis. The results confirmed the successful isolation of SSF. Human (hTERT) and sheep (sTERT) telomerase reverse transcriptase vectors were separately introduced into SSF, with cells passaged up to 36 generations following G418 selection. Microscopic examination and qRT-PCR results demonstrated that TERT transfection did not alter the morphology of SSF and led to stable, high levels of TERT expression (P < 0.01). Cell counting and flow cytometry revealed that TERT-transfected cells had higher viability and lower apoptosis rates compared to SSF (P < 0.05). Karyotype and soft agar colony formation assays indicated that hTERT and sTERT-transfected cells maintained normal characteristics without malignant transformation. β-galactosidase staining indicated that TERT transfection significantly reduced cellular senescence (P < 0.001). Additionally, sTERT-transfected cells exhibited higher TERT expression, enhanced viability, proliferation, and anti-senescence effects compared to hTERT-transfected cells (P < 0.05). In summary, the introduction of hTERT and sTERT effectively extends the lifespan of SSF, with sTERT demonstrating a more pronounced effect. This study provides critical evidence for preserving Mongolian sheep genetic resources and developing immortalized cell lines.

Insulin-like Growth Factor 2 mRNA-binding Protein 1 (IGF2BP1) is a candidate gene of significant interest for modulating economically important traits in livestock and poultry. The second intron of IGF2BP1 has been implicated in growth-related traits, though its precise mechanistic role remains elusive. Initial resequencing analyses in our laboratory indicated strong selective pressures on the IGF2BP1 genomic region, prompting the selection and identification of several single nucleotide polymorphisms (SNPs). Seven SNPs were mapped to the conserved region of the second intron, necessitating further investigation into their functional relevance and association with growth traits. In this study, 348 Nanjiang Yellow goats were analyzed, and the association analysis via the GLM program in SAS 9.4 identified five SNPs significantly correlated with growth traits. Notably, rs652062749(A > G) emerged as a critical locus influencing later-stage growth traits. Furthermore, strong linkage disequilibrium was observed among three SNPs, with the rs638185407 (T > A) variant markedly enhancing luciferase activity in H293T cells. Combination genotypes TTAACT, TTCCCC, and ATCACT were identified as superior for growth traits, offering theoretical insights for genetic co-breeding. This study underscores the potential utility of IGF2BP1 as a functional genetic marker in Nanjiang Yellow goat breeding programs.

This study aims to explore the effects of Yin and Yang Double Supplement Compound Chinese Medicine Preparations (YYSBFF) on the reproductive performance, antioxidant levels, and immunity of dairy goats. For the experiment, 36 Alps milk goats were selected and randomly divided into an experimental group and a control group, with 18 goats in each group. The ewes in the experimental group were fed with YYSBFF for 14 d prior to breeding and farrowing. The results of the experiment showed that the estrus rate, embryo acceptance rate, and lamb birth weight in the experimental group were higher than those in the control group, and the weak lamb rate was significantly lower. Moreover, the experimental group exhibited higher levels of reproductive hormones (FSH, LH), antioxidant factors (T-SOD, GSH-Px, MDA), and immunoglobulins (IgA, IgM, IgG) compared to the control group. There were no significant differences in liver and kidney function indicators (ALT, AST, TP, ALB, CREA, UREA) between the experimental and control groups (p > .05). These findings indicate that YYSBFF can enhance the reproductive performance of dairy goats by regulating the level of sex hormones, while also improving the body's antioxidant and immune abilities.

The study assessed the inclusion level effects of a phytogenic blend (PB) on performance and critical molecular biomarkers related to detoxification (Aryl hydrocarbon receptor; AhR) and antioxidant (Nuclear factor-erythroid 2-related factor 2; Nrf2) responses in layers' ovary and liver. Layers (n = 385; 21-week-old; Hy-Line Brown) were allotted to 5 treatments with 7 replicates of 11 hens each, for a 12-week feeding trial. Treatments were: control (CON) without PB or supplementation with PB at 250 (PB250), 750 (PB750), 1000 (PB1000) and 1500 mg/kg diet (PB1500), respectively. Performance was determined weekly for the 12-week experimental period. At the 6^th and 12^th experimental week ovarian and liver samples were collected for gene expression analysis. Increasing PB inclusion level, improved linearly and quadratically overall laying rate, egg mass and FCR, with optimal (P ≤ 0.001) performance notable in the PB750 compared to CON. The nutrigenomic analysis revealed that PB inclusion resulted (P ≤ 0.05) in consistent beneficial modulation of the AhR/Nrf2 pathway-related genes assessed at the 6^th and 12^th experimental week, both in the ovary and the liver of laying hens. In conclusion, PB beneficially modulated the ovarian and hepatic adaptive cytoprotection and supported the laying performance improvements recorded, with PB750 displaying the optimal benefits.

The natural flavonoid quercetin, which exhibits a range of biological activities, has been implicated in liver disease resistance in recent research. In vivo study attesting to quercetin's protective effect against metabolic-associated fatty liver disease (MAFLD) is inadequate, however. Here, our investigation explored the potential benefits of quercetin in preventing MAFLD in C57BL/6 mice fed a high-fat diet (HFD). The results revealed that quercetin ameliorated the aberrant enhancement of body and liver weight. The hepatic histological anomalie induced by MAFLD were also mitigated by quercetin. HFD-induced imbalance in serum LDL, HDL, AST, ALT, TG, and LDH was mitigated by quercetin. Mechanically, we found that quercetin improved lipid metabolism by reducing lipogenesis proteins including ACC, FASN, and SREBP-1c and enhancing β-oxidation proteins including PPARα and CPT1A. In vitro study demonstrated that quercetin regulated hepatic lipid metabolism by targeting SREBP-1c and PPARα. Additionally, quercetin enhanced the antioxidant capacity in HFD-treated mice by downregulating Nrf2 and HO-1 expressions and upregulating SOD and GPX1 expressions. The hyper-activation of inflammation was also restored by quercetin via eliminating the phosphorylation of IκBα and NF-κB p65. Collectively, our observations highlight that quercetin exerts hepatoprotective properties in MAFLD mice by regulating hepatic lipid metabolism, oxidative stress and inflammatory response.

Economy of Pakistan is heavily dependent upon agriculture and extensive use of pesticide is quiet common to enhance the crop yield. Imidacloprid is among the first choice pesticides in Pakistan and it has been reported that through run off along with water it ends up in water bodies affecting non target aquatic fauna. Through the present investigation, we are reporting the effects of Imidacloprid on the fatty acids composition of a non-target, commercially important carp: Labeo rohita. Fish were exposed to sub lethal concentration of Imidacloprid (120 mgL¹) for 2, 4 and 8 days (short term) as well as for 16, 32 and 64 days (long term experimental conditions). Pesticide untreated controls were also maintained for each treatment. Following the specific Imidacloprid exposure, fatty acid composition (%) was determined in the muscle of all experimental groups by using gas chromatography. Fish exposed to Imidacloprid for 8 days had reduced Palmitic acid (p = 0.02) and elevated muscle Arachidic acid (p < 0.001) than control group. Labeo rohita exposed to the pesticide for 32 days had elevated muscle Oleic (p = 0.02) and Linoleic acid (p = 0.02) while fish exposed to Imidacloprid to 64 days had reduced muscle Palmitic (p = 0.04) and Oleic acid (p = 0.03). In conclusion, we are reporting that the exposure to sub lethal concentration of Imidacloprid disturb the muscle fatty acid composition of Labeo rohita that may affect its food quality. The effects were more pronounced under long term experimental conditions and were probably due to potentiating lipid peroxidation and disturbed fish metabolism upon Imidacloprid exposure.