• Reproductive and Developmental Biology
• /
• 제38권3호
• /
• pp.123-127
• /
• 2014

This study was conducted for SNPs in the 5'-regions of estrogen receptor-${\alpha}$ (ESR-${\alpha}$), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genetic improvement was incurred low reproduction performance. The objective of this study was to investigate connections between single nucleotide polymorphisms (SNP) of Estrogen receptor-${\alpha}$ (ESR-${\alpha}$) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to genotype the cattle. The result showed two variants of the ESR-${\alpha}$ gene. The Bgl I cut the 492 bp amplification product into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction fragments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in these breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was $382.18{\pm}10.03$ days, and G/G was $381.69{\pm}5.22$ days. The SPC of A/G was $1.62{\pm}0.16$, and G/G was $1.32{\pm}0.04$. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was $7,253.00{\pm}936.00kg$ and of G/G was $8,747.51{\pm}204.88kg$, showing no significant difference.

• Asian-Australasian Journal of Animal Sciences
• /
• 제23권8호
• /
• pp.993-999
• /
• 2010

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

• 한국동물생명공학회지
• /
• 제34권2호
• /
• pp.117-122
• /
• 2019

The objective of this study was to establish an in vitro culture system for ovarian preantral follicles of B6D2F1. First, we optimized the in vitro preantral-follicle culture by culture duration, follicle stimulating hormone (FSH) type, and activin A concentration. Duration of in vitro culture for 9, 11, and 13 days was sufficient for the normal development of preantral follicles to antral follicles. Formation of cumulus cell-oocyte complex (COC) was induced by treatment with human chorionic gonadotropin (hCG; 2.5 IU/mL) and epidermal growth factor (EGF; 5 ng/mL). In addition, metaphase II (MII) oocytes formed during this in vitro culture of preantral follicles. In vitro preantralfollicle culture for 9 days showed higher rates of growth and maturation, thus yielding a greater number of antral follicles, and there were significant differences (p < 0.05) in the number of MII oocytes (that formed from these preantral follicles via differentiation) between the 9-day culture and 11-day or 13-day culture. The follicles cultured for 9 days contained a tightly packed well-defined COC, whereas in follicles cultured for 11 days, the COC was not well defined (spreading was observed in the culture dish); the follicles cultured for 13 days disintegrated and released the oocyte. Second, we compared the growth of the preantral follicles in vitro in the presence of various FSH types. There were no significant differences in the growth and maturation rates and in differentiation into MII oocytes during in vitro culture between preantral follicles supplemented with FSH from Merck and those supplemented with FSH from Sigma. To increase the efficiency of MII oocyte formation, the preantral follicles were cultured at different activin A concentrations (0 to 200 ng/mL). The control follicles, which were not treated with activin A, showed the highest rate of differentiation into antral follicles and into MII oocytes among all the groups (0 to 200 ng/mL). Therefore, activin A (50 to 200 ng/mL) had a negative effect on oocyte maturation. Thus, in this study, we propose an in vitro system of preantral-follicle culture that can serve as a therapeutic strategy for fertility preservation of human oocytes for assisted reproductive medicine, for conservation of endangered species, and for creation of superior breeds.

• Journal of Animal Science and Technology
• /
• 제57권3호
• /
• pp.8.1-8.8
• /
• 2015

Background: The demand for healthy, lean and consistent meat products containing low saturated fatty acid content and high quality polyunsaturated fatty acids (PUFA), especially long-chain (${\geq}C_{20}$) omega-3 PUFA, has increased in recent times. Fat deposition is altered by both the genetic background and dietary supplements, and this study aimed to assess the effect of dietary Spirulina supplementation levels on the mRNA expression patterns of genes controlling lipid metabolism in the subcutaneous adipose tissue (SAT) and Longissimus dorsi (ld) muscle of Australian crossbred sheep. Methods: Twenty-four weaned lambs belonging to four breeds under the same management conditions were maintained on ryegrass pasture and fed three levels of Spirulina supplement (control, low and high). In terms of nutrient composition, Spirulina is a nutrient-rich supplement that contains all essential amino acids, vitamins and minerals. It also is a rich source of carotenoids and fatty acids, especially gamma-linolenic acid (GLA) that infer health benefits. After slaughter, subcutaneous adipose tissue (SAT) and ld samples were subjected to mRNA extraction and reverse transcription using quantitative polymerase chain reaction (RT-qPCR) to assess the mRNA expression levels of the Aralkylamine N-acetyltransferase (AANAT), Adrenergic beta-3 receptor (ADRB3), B-cell translocation gene 2 (BTG2) and Fatty acid synthase (FASN) genes, which are associated with lipid metabolism. Results: Both low and high Spirulina supplementation levels strongly up-regulated the transcription of all the selected genes in both SAT and ld tissues (mostly in the subcutaneous adipose), but sheep breed and sex did not influence the gene expression patterns in these tissues. Conclusions: The evidence indicates that high Spirulina supplementation level resulted in a decrease in intramuscular fat content in Australian purebred and crossbred sheep due to the enhanced production of melatonin in sheep muscle tissues and strong up-regulation of mRNA expression of BTG2 in SAT which negatively affected fat deposition. In contrast, low Spirulina supplementation level strongly up-regulated the ADRB3 and FASN genes responsible for fat production. These findings are consistent with the observed phenotypic data suggesting that low Spirulina supplementation level can increase lamb production, with higher long-chain PUFA content.

• Journal of Animal Science and Technology
• /
• 제56권3호
• /
• pp.8.1-8.6
• /
• 2014

Background: Artificial insemination (AI) can serve as a powerful tool to the sheep owners for making rapid genetic progress of their flock. The AI in sheep is mostly performed using fresh semen with two reasons i) lambing rate following trans-cervical AI with frozen semen is limited by the inability of frozen-thawed sperm to transit the cervix and ii) the need of circumventing the cervical barrier through laparoscope aided intrauterine AI. Therefore, AI with frozen-thawed semen is not as widespread in sheep as it is in other domestic species. However, to get maximum benefits through the use of AI, frozen-thawed semen is a prerequisite because instead of high fertility, the short shelf life of fresh semen coupled with a limitation on the number of insemination doses achievable per unit time restricts the widespread use of individual sires. Therefore, in order to enhance lambing rate, a total of 240 trans-cervical artificial inseminations with frozen-thawed semen were performed in Bharat Merino ewes during autumn season either once in the evening (G-I, 10 h after onset of estrus, n = 100) or twice (G-II, 14 h and 22 h after onset of estrus, n = 140) i.e. once in the morning and again in the evening. Results: The pregnancy rate (proportion of pregnant ewes confirmed by ultrasonography at day 40) and lambing rate (proportion of ewes lambed) were higher in G-II as compared to G-I (26.4 vs 20% and 19.3 vs 10%, respectively). The difference in lambing rates was statistically (P < 0.05) significant. The depth of insemination within cervico-uterine tract had no significant effect on pregnancy and lambing rates. Conclusions: The results indicate that lambing rate in sheep following TCAI with frozen-thawed semen was significantly influenced by time of inseminations. Two inseminations after 14 and 22 h of onset of estrus enhanced the lambing rates of Bharat Merino sheep as compare to single insemination after 10 h of onset of estrus. The TCAI technique with frozen-thawed ram semen is promising and may serve as a valuable tool for genetic improvement of sheep breeds. Research efforts are going on worldwide to overcome the poor fertility following TCAI with frozen-thawed semen.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권5호
• /
• pp.627-632
• /
• 2007

FoxO1, FoxO3a and FoxO4 belong to the FoxO gene family, which play important roles in the PI3K/PKB pathway. In this study, we cloned the porcine FoxO1, FoxO3a and FoxO4 sequences and assigned them to SSC11p11-15, SSC1p13 and SSC xq13 using somatic cell hybrid panel (SCHP) and radiation hybrid panel (IMpRH). RT-PCR results showed that these three genes are expressed in multiple tissues. Sequencing of PCR products from different breeds identified a synonymous T/C polymorphism in exon 2 of FoxO3a. This FoxO3a single nucleotide polymorphism (SNP) can be detected by AvaII restriction enzyme. The allele frequencies of this SNP were investigated in Dahuabai, Meishan, Tongcheng, Yushan, Large White, and Duroc pigs. Association of the genotypes with growth and carcass traits showed that different genotypes of FoxO3a were associated with carcass length and backfat thickness between 6th and 7th ribs (BTR) and drip loss (p<0.05).

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권2호
• /
• pp.167-176
• /
• 2008

Galectins are a group of animal lectins consisting of galectin-type carbohydrate recognition domains (CRD) with relatively minor domains. The biological properties of galectins include the regulation of inflammation, intercellular adhesion, cell differentiation and cell death. The diverse kinds of galectin suggest variety in their biological roles. Galectin-1 is released during adipocyte differentiation and is associated with fat which is one of the important factors for meat quality. To verify expression level, a 0.5 kb clone of galectin-1 was obtained from cDNA prepared from back fat tissue of a Sancheong Berkshire pig with good quality meat, and the galectin-1 gene identified. The deduced amino acid sequence of the galectin-1 gene was compared with those obtained from other species. By using RT-PCR and Real time-PCR, an attempt was made to determine the expression level of galectin-1 and to compare with various tissues (tenderloin and back fat) taken from pigs in different groups. Grouping of pigs was based on growth-stage (weighing 60, 80, and 110 kg) and the sub-speciation (Yorkshire and Sancheong Berkshire pigs). We attempted to determine influences of pig species, growth stages and tissue variations on the expression level of the galectin-l gene and it was revealed that the expression pattern of the galectin-1 gene was significantly different (p<0.01 or p<0.05). Galectin-1 genes were expressed more highly in the back fat tissues of pigs weighing 110 kg than in those weighing 60 kg or 80 kg. However, the lowest expression was seen in the tenderloin tissues of pigs weighing 110 kg. Sancheong Berkshire pigs showed higher expression of the galectin-1 gene compared to Yorkshire pigs. Accordingly, it is considered that the expression pattern of the galectin-1 gene influences the growth of back fat tissues and the pig speciation relationship. Previous studies suggested that different expression of galectin-1 genes represents variety among the breeds and is closely related to fat tissue growth, conjugation and catabolism. Further, this study suggests that the expression of galectin-1 at a specific growth stage and tissue contributes significantly to the overall meat quality of Sancheong Berkshire pigs.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권2호
• /
• pp.245-251
• /
• 2008

The effects of feeding diets containing 20% (L) or 30% (H) neutral detergent fiber (NDF) (DM basis) on performance and carcass traits were studied in three breeds of pig, including pure Mong Cai (MC), crossbred Landrace$\times$Yorkshire (LY) and crossbred MC$\times$Yorkshire (F1). The experiment had a factorial design with two factors, breed and diet. Eighteen piglets of each breed ($60{\pm}3days$) were randomly allocated to three treatments: L-L, low fiber diet in both growing and finishing periods; L-H, low and high fiber diet in the growing and finishing period, respectively; and H-H, high fiber diet in both periods. The diets were iso-energetic and iso-nitrogenous within feeding period. The main fibrous ingredients of the diets were rice bran and cassava residue. There were no effects of fiber level on daily dry matter feed intake (DMI), expressed as g/kg metabolic body weight (BW0.75), in both feeding periods (p>0.05). DMI was highest for MC, followed by F1 and LY (p<0.001). Average daily gain (ADG) in L-L and L-H was higher than in H-H in the growing period (p<0.001) and overall (p<0.05), while feed conversion ratio (FCR) was higher in H-H than in L-L and L-H in the growing period (p<0.05) and overall, but no significant differences between treatments were found in the finishing period. In both periods, Landrace$\times$Yorkshire had the highest ADG and the lowest FCR, followed by F1 and Mong Cai (p<0.001). There were no interactions between breed and diet for performance and carcass traits. Carcass and dressing percentage was lower for L-H and H-H than for L-L (p<0.05). There were no significant differences among treatments in back fat thickness and lean meat percentage, or in crude protein and ether extract contents of lean meat. Carcass, dressing and lean meat percentage was highest for LY, lowest for MC and intermediate for F1 (p<0.001). It can be concluded that feeding a high fiber diet in the growing period reduced pig performance, but there was no effect in the finishing period. Pure Mong Cai pigs are not particularly suitable for meat purposes, although the F1 cross with Large White had reasonably good growth performance and carcass quality.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권3호
• /
• pp.424-435
• /
• 2020

Objective: The study was conducted to investigate variations in the immunophysiological responses to exercise-induced stress in Jeju and Thoroughbred horses. Methods: Blood samples were collected from the jugular veins of adult Jeju (n = 5) and Thoroughbred (n = 5) horses before and after 30 min of exercise. The hematological, biochemical, and immunological profiles of the blood samples were analyzed. Blood smears were stained and observed under a microscope. The concentration of cell-free (cf) DNA in the plasma was determined using real time polymerase chain reaction (PCR). Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells were separated using Polymorphprep, and the expression of various stress-related and chemokine receptor genes was measured using reverse transcriptase (RT) and real-time PCR. Results: After exercise, Jeju and Thoroughbred horses displayed stress responses with significantly increased rectal temperatures, cortisol levels, and muscle catabolism-associated metabolites. Red blood cell indices were significantly higher in Thoroughbred horses than in Jeju horses after exercise. In addition, exercise-induced stress triggered the formation of neutrophil extracellular traps (NETs) and reduced platelet counts in Jeju horses but not in Thoroughbred horses. Heat shock protein 72 and heat shock protein family A (Hsp70) member 6 expression is rapidly modulated in response to exercise-induced stress in the PBMCs of Jeju horses. The expression of CXC chemokine receptor 4 in PBMCs was higher in Thoroughbred horses than in Jeju horses after exercise. Conclusion: In summary, the different immunophysiological responses of Jeju and Thoroughbred horses explain the differences in the physiological and anatomical properties of the two breeds. The physiology of Thoroughbred horses makes them suitable for racing as they are less sensitive to exercise-induced stress compared to that of Jeju horses. This study provides a basis for investigating the link between exercise-induced stresses and the physiological alteration of horses. Hence, our findings show that some of assessed parameters could be used to determine the endurance performance of horses.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권2호
• /
• pp.187-196
• /
• 2020

Objective: Porcine respiratory disease is one of the most important health problems causing significant economic losses. To understand the genetic basis for susceptibility to swine enzootic pneumonia (EP) in pigs, we detected 102,809 single nucleotide polymorphisms in a total of 249 individuals based on genome-wide sequencing data. Methods: Genome comparison of susceptibility to swine EP in three pig breeds (Jinhua, Erhualian, and Meishan) with two western lines that are considered more resistant (Duroc and Landrace) using cross-population extended haplotype homozygosity and F-statistic (F_ST) statistical approaches identified 691 positively selected genes. Based on quantitative trait loci, gene ontology terms and literature search, we selected 14 candidate genes that have convincible biological functions associated with swine EP or human asthma. Results: Most of these genes were tested by several methods including transcription analysis and candidate genes association study. Among these genes: cytochrome P450 1A1 and catenin beta 1 (CTNNB1) are involved in fertility; transforming growth factor beta receptor 3 plays a role in meat quality traits; Wnt family member 2, CTNNB1 and transcription factor 7 take part in adipogenesis and fat deposition simultaneously; plasminogen activator, urokinase receptor (completely linked to AXL receptor tyrosine kinase, r² = 1) plays an essential role in the successful ovulation of matured oocytes in pigs; colipase like 2 (strongly linked to SAM pointed domain containing ETS transcription factor, r² = 0.848) is involved in male fertility. Conclusion: These adverse genes susceptible to swine EP may be selected while selecting for economic traits (especially reproduction traits) due to pleiotropic and hitchhiking effect of linked genes. Our study provided a completely new point of view to understand the genetic basis for susceptibility or resistance to swine EP in pigs thereby, provides insight for designing sustainable breed selection programs. Finally, the candidate genes are crucial due to their potential roles in respiratory diseases in a large number of species, including human.