Growth factors improve the proliferation of Jeju black pig muscle cells by regulating myogenic differentiation 1 and growth-related genes

Park, Jinryong,Lee, Jeongeun,Song, Ki-Duk,Kim, Sung-Jo,Kim, Dae Cheol,Lee, Sang Cheol,Son, Young June,Choi, Hyun Woo,Shim, Kwanseob 아세아태평양축산학회 2021 Animal Bioscience Vol.34 No.8

Objective: The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells. Methods: Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5×10⁵ cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (MyoD) expression and growth-related gene expression in JBP muscle cells, respectively. Results: We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (EGF+bFGF) than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of collagen type XXI alpha 1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells. Conclusion: Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.

Effect of Buffalo Follicular Fluid Alone and in Combination with PMSG and M199 on in vitro Buffalo Oocyte Maturation

Gupta, P.S.P.,Nandi, S.,Ravindranatha, B.M.,Sarma, P.V. 아세아태평양축산학회 2001 Animal Bioscience Vol.14 No.5

The effect of replacement of in vitro maturation medium completely with the buffalo follicular fluid (buFF) on in vitro oocyte maturation of buffalo oocytes was studied. 5 to 8 buffalo cumulus oocyte complexes were cultured in a single drop with each of the eight media studied i.e., M199+steer serum (10% v/v), M199+steer serum (10% v/v)+PMSG, M199+buFF (10% v/v), M199+buFF (10% v/v)+PMSG, M199+buFF (50% v/v), M199+buFF (50% v/v)+ PMSG, buFF (100%) and buFF+PMSG at $39^{\circ}C$ and 5% $CO_2$ in air for 24 h. Supplementation of M199 with Steer serum alone resulted in IVM rate of 35% only. When the above medium was supplemented with PMSG, the maturation rate rallied to 82%. Significant increase in the maturation rates were observed when M199 was supplemented with increasing levels of buFF. A further increase in the maturation rate was also obtained when PMSG was incorporated into the medium of M199 supplemented with buFF. The rate of maturation was to the tune of 91% when oocytes were matured in buFF alone which was increased non significantly on the addition of PMSG. Highest maturation rate (97%) obtained with M199+buFF (50%v/v)+PMSG did not differ significantly from that obtained by either M199+buFF (10%v/v)+PMSG or buFF+PMSG. It is suggested that buFF alone without any supplementation can form the effective in vitro maturation medium for buffalo oocytes.

Induction of Ski Protein Expression upon Luteinization in Rat Granulosa Cells

Kim, Hyun,Kim, Dong-Hun,Park, Soo-Bong,Ko, Yeoung-Gyu,Kim, Sung-Woo,Do, Yoon-Jun,Park, Jae-Hong,Yang, Boh-Suk 아세아태평양축산학회 2012 Animal Bioscience Vol.25 No.5

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells; however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to locate Ski protein in the rat ovary during luteinizationto predict the possible role of Ski. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rats, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of the corpus luteum (CL). Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggests that its expression is regulated post-transcriptionally.

Protein molecular structure, degradation and availability of canola, rapeseed and soybean meals in dairy cattle diets

Yujia Tian,Xuewei Zhang,Rongcai Huang,PeiqiangYu 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.9

Objective: The aims of this study were to reveal the magnitude of the differences in protein structures at a cellular level as well as protein utilization and availability among soybean meal (SBM), canola meal (CM), and rapeseed meal (RSM) as feedstocks in China. Methods: Experiments were designed to compare the three different types of feedstocks in terms of: i) protein chemical profiles; ii) protein fractions partitioned according to Cornell Net Carbohydrate and Protein System; iii) protein molecular structures and protein second structures; iv) special protein compounds-amino acid (AA); v) total digestible protein and energy values; vi) in situ rumen protein degradability and intestinal digestibility. The protein second structures were measured using FT/IR molecular spectroscopy technique. A summary chemical approach in National Research Council (NRC) model was applied to analyze truly digestible protein. Results: The results showed significant differences in both protein nutritional profiles and protein structure parameters in terms of α-helix, β-sheet spectral intensity and their ratio, and amide I, amide II spectral intensity and their ratio among SBM, CM, and RSM. SBM had higher crude protein (CP) and AA content than CM and RSM. For dry matter (DM), SBM, and CM had a higher DM content compared with RSM (p<0.05), whereas no statistical significance was found between SBM and CM (p = 0.28). Effective degradability of CP and DM did not demonstrate significant differences among the three groups (p>0.05). Intestinal digestibility of rumen undegradable protein measured by three-step in vitro method showed that there was significant difference (p = 0.05) among SBM, CM, and RSM, which SBM was the highest and RSM was the lowest with CM in between. NRC modeling results showed that digestible CP content in SBM was significantly higher than that of CM and RSM (p<0.05). Conclusion: This study suggested that SBM and CM contained similar protein value and availability for dairy cattle, while RSM had the lowest protein quality and utilization.

Effects of a specific blend of essential oils on apparent nutrient digestion, rumen fermentation and rumen microbial populations in sheep fed a 50:50 alfalfa hay:concentrate diet

N. Khateri,O. Azizi,Jahani-Azizabadi 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.3

Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.

Newborn traits associated with pre-weaning growth and survival in piglets

Morakot Nuntapaitoon,Ramon Muns,Padet Tummaruk 아세아·태평양축산학회 2018 Animal Bioscience Vol.31 No.2

Objective: Piglet pre-weaning mortality is an important variable indicating the efficacy of farrowing management and animal well-being during lactation. The present study determined the association of newborn traits measured soon after birth with piglet pre-weaning mortality and growth. Methods: In total, 805 piglets born from 57 multiparous sows were investigated. Their blood oxygen saturation, blood glucose and rectal temperature at 24 h after birth (RT24h) were monitored. Birth order, sex, skin color, integrity of the umbilical cord, attempts to stand and birth intervention were monitored. Piglets were weighed at day 0, 7, and 21 to evaluate average daily gain (ADG). Results: Piglet pre-weaning mortality for lactation period was 12.6% and cumulative mortality during the first 7 days of age was 8.6%. A higher proportion of piglets with pale skin color died compared to piglets with normal skin color (26.7% vs 7.7%, p<0.001). A higher (p<0.001) proportion of piglets that attempted to stand after 5 min (38.5%) died compared to piglets that attempted to stand within 1 min (6.3%) after birth. Piglet body weight at birth (BWB), blood glucose and the number of piglets born alive (BA) were correlated with ADG (p<0.05). Piglets with BWB <1.30 kg had higher (p<0.001) mortality rate than piglets with BWB≥1.80 kg (19.0% vs 3.3%) and piglets with BWB 1.30 to 1.79 kg (4.0%). Piglet with RT24h <37.0°C had higher (p<0.001) mortality rate (86.2%) than piglets with RT24h >38.5°C (3.9%). Conclusion: Low BWB and low RT24h compromise piglet survival during the lactation period in the tropical conditions. Piglets in the litters with a high BA, low BWB and low blood glucose have reduced ADG.

Effects of season and single layer centrifugation on bull sperm quality in Thailand

Nongbua Thanapol,Utta Apirak,Am-in Nutthee,Suwimonteerabutr Junpen,Johannisson Anders,Morrell Jane M 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.9

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

Dietary supplementation with different types of fiber in gestation and lactation: effects on sow serum biochemical values and performance

Weng Ruey-Chee 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.8

Objective: Three types of dietary fiber were fed to sows during gestation and lactation stages to monitor their physiological and metabolic adaptations during the pre-partum period and to determine how these effects may influence the lactation period and sow performance. Methods: Soon after breeding, 54 sows were selected and were fed with 20% supplementation as fed of wheat bran (WB), soya hulls (SH), or rice hulls (RH) in diets during gestation and lactation. Sows were weighed, backfat thickness was measured ultrasonically and jugular blood samples were collected from all sows. The litter size was equalized to 10, by fostering piglets from sows on the same treatment. Results: Sows gained 22.0, 21.8, and 25.5 kg of net maternal body weight during gestation (for WB, SH, and RH sows, respectively; p = 0.007). There was no treatment effect on the body weight change during lactation (p = 0.158), however RH sows consumed an average of 133.66 kg of feed, WB sows took 121.29 kg and SH sows took 126.77 kg during lactation (p<0.001). The SH litters gained an average of 59.34 kg of weight during lactation, while other litters gained 51.58 and 49.98 kg (for WB and RH litters, respectively; p<0.001). Exception for aspartate aminotransferase and alanine aminotransferase, measured serum biochemical values were broadly in agreement with earlier reports. Despite the use of additional vegetable oil to balance the energy level, RH sows still had lower concentrations of serum triglycerides in late gestation. Conclusion: Different types of fibrous ingredients in the gestation diet influenced most of the investigated reference values for sows. The values of serum biochemical parameters were generally not affected by fiber type during the lactation stage. The SH supplementation for sows is an effective approach to give heavier litters at birth and weaning and to increase voluntary feed intake in early lactation.

Evaluation of Antimicrobial Activities of Sequential Spray Applications of Decontamination Treatments on Chicken Carcasses

Hakan Benli,Marcos X. Sanchez-Plata,Osman Irfan Ilhak,Maryuri T. Nunez De Gonzalez,Jimmy T. Keeton 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.3

The objective of this study was to evaluate the effects of sequential applications of ε-polylysine (EPL) or lauramide arginine ethyl ester (LAE) sprays followed by an acidic calcium sulfate (ACS) spray on inoculated chicken carcasses to reduce Salmonella (Salmonella enterica serovars including Salmonella typhimurium and Salmonella enteritidis) contamination during 6 days of storage (4.4°C). Secondly, reductions of the resident microflora were studied on uninoculated chicken carcasses following the sequential application of the treatments, chilling and 10 days of storage at 4.4°C. The treatment of Salmonella inoculated carcasses with 300 mg/L EPL followed by 30% ACS (EPL300-ACS30) sprays reduced Salmonella counts initially by 1.5 log cfu/mL and then by 1.2 log cfu/mL (p<0.05) following 6 days of storage at 4.4°C. Likewise, 200 mg/L LAE followed by 30% ACS (LAE200-ACS30) treatment reduced initial Salmonella counts on poultry carcasses by 1.8, 1.4 and 1.8 log cfu/mL (p<0.05), respectively, after 0, 3, and 6 days storage. Immediately after the treatments, EPL300-ACS30 and LAE200-ACS30 both reduced Escherichia coli counts significantly by 2.6 and 2.9 log cfu/mL, respectively. EPL300-ACS30 and LAE200-ASC30 were effective in lowering psychrotroph counts by 1 log cfu/mL on day 10 when compared to the control and distilled water treatments. This study demonstrated that EPL300-ACS30 and LAE200-ACS30 were effective in reducing Salmonella on inoculated chicken carcasses both after treatment and during the storage at 4.4°C for up to 6 days. In addition, reductions in psychrotroph counts indicated that these treatments might have the potential to increase the shelf-life of poultry carcasses.

Correlations among Stress Parameters, Meat and Carcass Quality Parameters in Pigs

Marija Dakmanovic,Milan Z. Baltic,Jelena Duric,Jelena Ivanovic,Ljuba Popovic,Milica Todorovic,Radmila Markovic,Srdan Pantic 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.3

Relationships among different stress parameters (lairage time and blood level of lactate and cortisol), meat quality parameters (initial and ultimate pH value, temperature, drip loss, sensory and instrumental colour, marbling) and carcass quality parameters (degree of rigor mortis and skin damages, hot carcass weight, carcass fat thickness, meatiness) were determined in pigs (n = 100) using Pearson correlations. After longer lairage, blood lactate (p<0.05) and degree of injuries (p<0.001) increased, meat became darker (p<0.001), while drip loss decreased (p<0.05). Higher lactate was associated with lower initial pH value (p<0.01), higher temperature (p<0.001) and skin blemishes score (p<0.05) and more developed rigor mortis (p<0.05), suggesting that lactate could be a predictor of both meat quality and the level of preslaughter stress. Cortisol affected carcass quality, so higher levels of cortisol were associated with increased hot carcass weight, carcass fat thickness on the back and at the sacrum and marbling, but also with decreased meatiness. The most important meat quality parameters (pH and temperature after 60 minutes) deteriorated when blood lactate concentration was above 12 mmol/L.