• Asian-Australasian Journal of Animal Sciences
• /
• 제32권6호
• /
• pp.757-766
• /
• 2019

Objective: MicroRNAs are a class of endogenous small regulatory RNAs that regulate cell proliferation, differentiation and apoptosis. Recent studies on miRNAs are mainly focused on mice, human and pig. However, the studies on miRNAs in skeletal muscle of sheep are not comprehensive. Methods: RNA-seq technology was used to perform genomic analysis of miRNAs in prenatal and postnatal skeletal muscle of sheep. Targeted genes were predicted using miRanda software and miRNA-mRNA interactions were verified by quantitative real-time polymerase chain reaction. To further investigate the function of miRNAs, candidate targeted genes were enriched for analysis using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment. Results: The results showed total of 1,086 known miRNAs and 40 new candidate miRNAs were detected in prenatal and postnatal skeletal muscle of sheep. In addition, 345 miRNAs (151 up-regulated, 94 down-regulated) were differentially expressed. Moreover, miRanda software was performed to predict targeted genes of miRNAs, resulting in a total of 2,833 predicted targets, especially miR-381 which targeted multiple muscle-related mRNAs. Furthermore, GO and KEGG pathway analysis confirmed that targeted genes of miRNAs were involved in development of skeletal muscles. Conclusion: This study supplements the miRNA database of sheep, which provides valuable information for further study of the biological function of miRNAs in sheep skeletal muscle.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권7호
• /
• pp.928-935
• /
• 2008

The myostatin gene of seven important meat (Beltex (Australia), Beltex$\times$Huyang (F1), Meat and Multi-Prolific Chinese Merino Fine Wool, Meat Chinese Merino Fine Wool and Dorper (South Africa)) and non-meat (Huyang and Kazak) sheep breeds was analyzed to study the genetic basis of muscular hypertrophy (double muscling) phenotype in sheep. SNPs, four in regulatory regions and several in the introns in the myostatin gene, were identified, and the former four SNPs were used for further studies. Twelve haplotypes were predicted by PHASE program, of which four main haplotypes (1, 3, 7, 9) were present in 90% of the 364 sheep in the study. Haplotypes 1-4 were mainly present in meat breeds while haplotypes 7 and 9 dominated the non-meat breeds. The association between haplotypes and average daily gain (ADG) was analyzed among 116 sheep with production data, Haplo2 (CGAA) and Haplo8 (TGAA) were identified to have significant (p<0.05) effect on ADG by the model (JMP5.1 software) taking into account the effects of breed, family background, haplotype, birth weight and sex. ADG of these haplotype groups also correlated well (r = 0.82) with hypertrophic phenotype scores. In conclusion, the mutations -956 (T$\rightarrow$C), -41 (C$\rightarrow$A) and 6223 (G$\rightarrow$A) involved in Haplo2 and 8 may be associated with the double-muscling trait by influencing myostatin function and be suitable markers in selecting meat sheep.

• Asian-Australasian Journal of Animal Sciences
• /
• 제32권10호
• /
• pp.1540-1547
• /
• 2019

Objective: This study was conducted to evaluate the effects of tannins and cellulase on growth performance, nutrient digestibility, blood profiles, intestinal morphology, and carcass characteristics in Hu sheep. Methods: A total of 48 three-month-old meat Hu sheep ($25.05{\pm}0.9kg$) were blocked based on body weight, and randomly allotted to 4 treatments with 3 replicates of 4 sheep each. The experiment lasted for 80 d, and dietary treatments were as follows: i) CON, control diet; ii) TAN, CON+0.1% tannins; iii) CEL, CON+0.1% cellulase; iv) TAN+CEL, CON+0.1% tannins and 0.1% cellulase. Results: Compared with CON, CEL, and TAN+CEL had greater (p<0.05) final body weight (FBW) and average daily gain but lower (p<0.05) feed conversion ratio, while FBW of TAN+CEL was lower (p<0.05) than that of CEL. The apparent total tract digestibility (ATTD) of dry matter in TAN, CEL, and TAN+CEL groups were higher (p<0.05) than that in CON. CEL and TAN+CEL groups had greater (p<0.05) ATTD of crude fiber compared with TAN and CON, while TAN group had lower (p<0.05) ATTD of crude protein than other treatments. TAN, CEL, and TAN+CEL groups increased (p<0.05) serum globulin and alkaline phosphatase but decreased (p<0.05) albumin/globulin. Serum total protein was greatest for TAN+CEL, intermediate for TAN and CEL and least for CON (p<0.05). TAN+CEL group increased (p<0.05) dressing percentage compared with CON, while the backfat thickness of CEL was lower (p<0.05) than that of CON. The villus height of jejunum and ileum in CEL and TAN+CEL groups were greater (p<0.05) than that in CON, and the crypt depth and villus height: crypt depth of jejunum were increased (p<0.05) in TAN, CEL, and TAN+CEL groups. Conclusion: The addition of tannins and cellulase together promoted nutrient digestion, liver protein synthesis and intestinal development and thus improved growth performance and carcass characteristics.

• Asian-Australasian Journal of Animal Sciences
• /
• 제29권3호
• /
• pp.413-418
• /
• 2016

Myostatin (MSTN) is a secreted growth factor expressed in skeletal muscle and adipose tissue that negatively regulates skeletal muscle mass. Gene knockout of MSTN can result in increasing muscle mass in sheep. The objectives were to investigate whether myostatin gene can be edited in sheep by transcription activator-like effector nucleases (TALENs) in tandem with single-stranded DNA oligonucleotides (ssODNs). We designed a pair of TALENs to target a highly conserved sequence in the coding region of the sheep MSTN gene. The activity of the TALENs was verified by using luciferase single-strand annealing reporter assay in HEK 293T cell line. Co-transfection of TALENs and ssODNs oligonucleotides induced precise gene editing of myostatin gene in sheep primary fibroblasts. MSTN gene-edited cells were successfully used as nuclear donors for generating cloned embryos. TALENs combined with ssDNA oligonucleotides provide a useful approach for precise gene modification in livestock animals.

• Asian-Australasian Journal of Animal Sciences
• /
• 제31권10호
• /
• pp.1550-1557
• /
• 2018

Objective: Circular RNAs (circRNAs) are a newfound class of non-coding RNA in animals and plants. Recent studies have revealed that circRNAs play important roles in cell proliferation, differentiation, autophagy and apoptosis during development. However, there are few reports about muscle development-related circRNAs in livestock. Methods: RNA sequencing analysis was employed to identify and annotate circRNAs from longissimus dorsi of sheep. Reverse transcription followed by real-time quantitative (q) polymerase chain reaction (PCR) analysis verified the presence of these circRNAs. Targetscan7.0 and miRanda were used to analyse the interaction of circRNA-microRNA (miRNA). To investigate the function of circRNAs, an experiment was conducted to perform enrichment analysis hosting genes of circRNAs using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Results: About 75.5 million sequences were obtained from RNA libraries of sheep skeletal muscle. These sequences were mapped to 729 genes in the sheep reference genome. We identified 886 circRNAs, including numerous circular intronic RNAs and exonic circRNAs. Reverse transcription PCR (RT-PCR) and DNA sequencing analysis confirmed the presence of several circRNAs. Real-Time RT-PCR analysis exhibited resistance of sheep circRNAs to RNase R digestion. We found that many circRNAs interacted with muscle-specific miRNAs involved in growth and development of muscle, especially circ776. The GO and KEGG enrichment analysis showed that hosting genes of circRNAs was involved in muscle cell development and signaling pathway. Conclusion: The study provides comprehensive expression profiles of circRNAs in sheep skeletal muscle. Our study offers a large number of circRNAs to facilitate a better understanding of their roles in muscle growth. Meanwhile, we suggested that circ776 could be analyzed in future study.

• Asian-Australasian Journal of Animal Sciences
• /
• 제6권4호
• /
• pp.601-605
• /
• 1993

This study was purposed to investigate the efficiency of protein utilization of rapeseed meal (RSM) and formaldehyde-treated RSM (TRSM) by sheep, and their influence on performance of growing heifers. Experiment 1 was conducted according to a double $3{\times}3$ Latin square design involving sex yearling Hu Sheep and three experimental diets. All diets contained 600 g ammoniated rice straw and 200 g concentrate mixture. Three dietary treatments were: (1) 100 g RSM + 40 g soybean meal (URUS), (2) 100 g TRSM + 40 g soybean meal (TRUS) and (3) 100 g TRSM + 40 g treated soybean meal (TRTS). Apparent nitrogen digestibility was significantly higher for diet TRUS than that for URUS or TRTS (p<0.05), but with no significant difference between URUS and TRTS (p>0.05). Proportion of nitrogen retention (NR) to the digestible nitrogen intake for diet TRUS and TRTS was 25.57 (p<0.05) and 23.44% (p<0.05) higher than that for URUS respectively. As a result, proportion of NR to nitrogen intake for diet TRUS and TRTS was 34.74 (p<0.05) and 23.78% (p<0.05) higher than that for URUS respectively. Experiment 2 was conducted with 59 Holstein heifers. They were 12-20 months of age at the start of the trial. The experiment was a $2{\times}2$ factorial trial in which the heifers were given the ammoniated rice straw ad libitum and 1.5 kg hay, and supplemented with either RSM or TRSM at a daily allowance of 1.2 or 1.8 kg per day. The live weight gains for heifers receiving 1.2 and 1.8 kg/d of RSM or TRSM were 0.491 and 0.556 or 0.564 and 0.665 kg/d, respectively. The results suggest that formaldehyde treatment can effectively improve the efficiency of protein utilization of rapessed meal and cattle's performance.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권8호
• /
• pp.1241-1245
• /
• 1999

Nine two-year old sheep fitted with rumen and duodenum cannulas were used to study the effect of controlling protozoa flora on the degradation and utilization of dietary fibre and protein in the rumen and on nitrogenous flow to the duodenum. There were three groups in this experiment: defaunation (DF); partial defaunation (PDF); faunation (F) as control. Results showed that: 1,There were no differences between treatments in dietary DM degradation in the rumen, but defaunation and partial defaunation increased the quantity of nitrogenous material in the rumen and the flow of N to duodenum. 2, partial defaunation and defaunation improved the degradabilities of dietary NDF, ADF and HC, but there were no differences between the defaunated and partially defaunated groups. 3, Partial defaunation decreased the degradability of dietary protein in the rumen. There was no difference between defaunated and faunated groups. 4, Defaunation and partial defaunation increased the quantity of total N (TN) and microbial N (MCN) in the rumen and the amounts entering the duodenum. The protozoa N (PN) flow in the faunated group was higher than that in the partially defaunated group, and the amino acid pattern in the digesta at the proximal duodenum in the defaunated group was closer to the ideal amino acid pattern. 5, There were differences in the mole percent of acetic, propionic, total-VFAs and the non-glucogenic to glucogenic VFAs ratio (NGR) value in the rumen fluids. The order was as follows: mole percent of acetate: F>PDF>DF; mole percent of propionate: DF>PDF>F; total-VFAs: PDF>F>DF; NGR: F>PDF>DF.

• Asian-Australasian Journal of Animal Sciences
• /
• 제15권11호
• /
• pp.1591-1598
• /
• 2002

The effect of different dietary structural carbohydrate (SC) to nonstructural carbohydrate (NSC) ratios on fiber degradation, digestion, flow, apparent digestibility and rumen fluid characteristics was studied with a design using 18 wethers fitted with permanent rumen and duodenum cannulae. All sheep were divided into six groups randomly, receiving six diets with varying SC to NSC ratios. All diets contained the same proportion of wheat straw and concentrate. The dietary SC to NSC ratios were adjusted by adding cornstarch to the concentrate supplements. The duodenal and fecal flows of dry matter (DM), neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose (HC) and cellulose (CEL) were estimated using chromium-mordanted wheat straw as a flow marker. The degradation parameters of wheat straw DM, NDF, ADF, HC and CEL were determined by incubating the ground wheat straw in nylon bags in the rumen for different periods of time. There was no effect (p>0.05) of the different dietary SC to NSC ratios on rumen pH or $NH_3$-N, but acetate, propionate and butyrate concentrations were significantly affected (p<0.05 or p<0.01) by dietary SC to NSC ratios in the rumen fluid. When the dietary SC to NSC ratio was 2.86, the highest rumen degradability of wheat straw DM, NDF, ADF and CEL was found, but the highest apparent rumen digestibilities of DM, NDF, ADF, HC and CEL occurred at a 2.64 SC to NSC ratio. However, because of compensatory digestion in the hindgut, the apparent digestibilities of DM, NDF, ADF, HC and CEL were highest when the dietary SC to NSC ratio was 2.40. In conclusion, there is a optimal range of dietary SC to NSC ratios (between 2.86 and 2.40) that is beneficial to maximize wheat straw fiber degradation and apparent digestibility.

• Asian-Australasian Journal of Animal Sciences
• /
• 제14권10호
• /
• pp.1374-1382
• /
• 2001

Fifteen Inner Mongolian wethers with permanent ruminal and duodenal cannulas were used to study the effects of dietary rumen-undegradable protein (RUP) to rumen-degradable protein (RDP) ratios or protein sources on fiber digestion in the gastrointestinal tract and ruminal fluid characteristics. Fiber digestion and ruminal fermentation were not affected (p>0.05) by dietary RUP to RDP ratios (from 1.54 to 0.72). Soybean meal supplementation improved ruminal digestion. Fish meal supplementation increased (p<0.05) the ruminal degradability of fiber. The different RUP to RDP ratios (from 1.54 to 0.72) did not influence (p>0.05) ruminal fluid pH, but there were differences (p<0.05) in ruminal fluid $NH_3-N$ concentration because of urea replacement. Soybean meal as a dietary protein source decreased (p<0.05) ruminal fluid pH and increased (p<0.05 or p<0.01) $NH_3-N$, acetate, propionate and butyrate concentrations in the rumen. Fish meal as a dietary protein source decreased (p<0.05 or p<0.01) ruminal $NH_3-N$ and acetate concentrations and increased (p<0.05) ruminal propionate concentration. It can be concluded that dietary protein sources have more significant effect on fiber digestion and ruminal fermentation than different dietary RUP to RDP ratios, when the dietary crude protein requirements of growing sheep are satisfied.

• Asian-Australasian Journal of Animal Sciences
• /
• 제11권4호
• /
• pp.455-461
• /
• 1998

This study was proposed to investigate effects of inclusion of Chinese milk vetch silage (MVS) and rapeseed meal (RSM) on the growth and rumen fermentation of Hu-sheep. Fifty weanling lambs were randomly divided into five equal groups and offered ammoniated rice straw (ABRS) ad libitum along with 100 g concentrate (Trial 1). The animals in $T_0$, $T_1$, $T_2$, $T_3$ and $T_4$ group were respectively supplemented with MVS at levels of 0, 0, 7, 14 or 21% and with RSM at levels of 0, 15, 10, 5 or 0%. Daily gain of lambs was significantly (p<0.05) higher in $T_1$, $T_2$ and $T_3$ group than that in $T_0$ and $T_4$ group. Feed conversion ratio was greatly reduced in supplemented groups as compared with $T_0$ group. In trial 2, five sheep with rumen cannulae were used in a $5{\times}5$ Latin square design. The experimental treatments were as described in Trial 1, but without concentrate. The intake of AVRS was significantly (p<0.05) lower in $T_4$ group than that in $T_0$ group, and also significantly (p<0.05) lower than those in $T_1$ and $T_2$ group. Little difference among all treatments was found in 48h DM degradability of ABRS, MVS and RSM, and in rumen pH value and microbial protein concentration. Rumen concentrations of individual and total VFA tended to be higher in supplemented groups than those in $T_0$ group.These rusults suggest that supplementation with RSM or RSM plus MVS can effectively improve the performance of lambs, and may fail to influence markedly the rumen digestion of ABRS and rumen environments.