• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.51-61
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• 2001

This study was conducted to determine developmental ability of reconstructed embryos by nuclear transfer using somatic cell of Korean bull with high genetic value. Fibroblast cells obtained from ear biopsy of the bull were cultured in Dulbecco's Modified Eagle's medium (DMEM) at 37$^{\circ}C$ in air containing 5% $CO_2$. The cummulus-oocyte complexes were collected from slaughterhouse and were matured in vitro for 20 h in TCM 199 culture medium and the oocytes were then enucleated in modified phosphate buffered saline with cytochalasin B. Matured bovine oocytes were enucleated by aspirating the first polar body and metaphase chromatin using a beveled pipette in modified phosphate buffered saline. The ear fibroblast cells were fused into enucleated oocyte by electrical stimulation. The reconstructed oocytes were activated with ionomycine and 6-dimethylaminopurine, and then cultured in CR1aa medium for 7.5 days. Out of 524 bovine eggs reconstructed by nuclear transfer 65.6%(277/422) embryos were cleaved, and 30.7% (85/277) cleaved embryos were developed to the morula to blastocysts. There was no difference of developmental ability in vitro of reconstructed embryos regardless of donor cell passages. In order to determine fate of foreign mitochondria of donor nucleus, the Mito Tracker stained cells were fused into enucleated oocytes. The donor mitochondria were detected early stage of embryos, but disappeared rapidly. The developmental ability of reconstructed embryos was not impaired by Mito Tracker treatments. The results indicate that viable reconstructed embryos can be producted by nuclear transfer using somatic cell of Korean bulls.bulls.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.259-269
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• 1996

The present study was undertaken to examine the critical effect of $Ca^2$+ concentration on electrostimulation and post-electrostimulation media for electric activation of in vitro matured oocytes of Korean Native Cattle. Oocytes collected from slaughterhouse ovaries were matured in TCM 199 containing FSH, estradiol-17$\beta$ and FBS with granulosa cell monolayer for 24 hours and denuded with hyaluronidase. And then cumulus-free oocytes were submitted to a DC field of 1.0 kV/cm for 60 $\mu$sec in electroporation media(0.28 M mannito' and PBS) with different $Ca^2$+ concentations (0.00, 0.05, 0.10 and 0.15 mM). Stimulated oocytes were stained and examined for pronuclear formation after incuhation in SOF for 12 hours. The rates of pronuclear formation in hovine oocytes electrically stimulated in 0.28 M mannitol with 0.05, 0.10 and 0.15 mM $Ca^2$+(60.3, 82.2 and 75.0%) were significantly higher than without $Ca^2$+(6.3%) at 12 hours after an electric pulse(p<0.005). The activation rates of Korean Native Cattle oocytes stimulated in PBS supplemented with 0.05, 0.10 and 0.15 mM $Ca^2$+(71.0, 75.8 and 75.4%) were significantly higher than without $Ca^2$+(23.5%) after post-stimulation incubation(p<0.005). After incubation of oocytes in SOF with and without $Ca^2$+ following electric stimulation in 0.28 M mannitol with 0.10 mM $Ca^2$+, the rates of pronuclear formation of bovine oocytes in $Ca^2$+-free SOF(85.7%) was significantly higher than in SOF with 1.71 mM $Ca^2$+(62.5%, p<0.05). When oocytes were stimulated in two electrostimulation media supplemented with $Ca^2$+ and incubated in $Ca^2$+-free SOF, there were no significant differences in the rates of pronuclear formation hetween 0.28 M mannitol and PBS. These results indicate that a single electric pulse could induce activation of Korea Native Cattle oocytes in 0.28 M mannitol and PBS supplemented with $Ca^2$+. Furthermore, to improve the activation rates, it was hetter that stimulated oocytes were incubated in $Ca^2$+-free SOF after electric stimulation than in SOF with $Ca^2$+.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.760-768
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• 2004

The present study was undertaken to evaluate the accuracy of ultrasonography in early pregnancy diagnosis in goats. Ultrasonographic scanning with real time B-mode ultrasound machine having 5 MHz linear array transducer was performed on gravid uterus (n=24) obtained from slaughterhouse (Group I). Crown rump length (CRL) measured by ultrasound was found significantly different (p<0.05) with actual CRL measured after dissection in early pregnancy. However, age predicted by ultrasound through the measurement of CRL was found highly correlated (r=0.92) with age measured after dissection through CRL and the weight of fetus. Ages predicted by ultrasound through the measurement of trunk diameter (TD) and uterine diameter (UD) and ages measured after dissection were found highly and equally correlated (r=0.98) and did not differ significantly. Data from six does synchronized (Group II) with PGF2$\alpha$ (Estrumate) at 11 days apart were collected through ultrasound from 17 to 42 days post breding. The correlation between CRL and gestational age was high (r=0.97) in day 30 to 42 post breeding. A high coefficient of correlation (r=0.98) was also observed between predicated age by ultrasound and actual age calculated after kidding. The correlation between CRL and gestational age by the formula Y=(a+bX) i.e. Y=24.42+0.39 X where Y=gestational age and X=CRL, was recorded very high (r=0.99). Accuracy of ultrasonography was lowest on day 17 to 19 (66%) and reached 100% on day 34. Data from 30 does (group III) randomly subjected to only one time ultrasounds scanning to assess the accuracy of pregnancy diagnosis were also obtained. Ages predicted by TD and UD measurements were observed to be non-significantly different with actual age obtained after kidding and correlation between ages predicted by TD and UD measurement with actual age after kidding was found equally and highly correlated (r=0.98). The operator's accuracy in the whole experiment including all three groups was found to be 92%. The sensitivity was 93% and specificity was 86%. From the present study, it was observed that CRL was the most reliable parameter to find out gestational age in early pregnancy and the new formula derived was found very accurate to find out gestational age. TD and UD were also found to be equally reliable parameter to find out gestational age in mid and late stage of pregnancy through ultrasonography. It was concluded that ultrasonography by real time B mode with 5 MHz transrectal transducer was found to be reliable, safe and accurate and practicable means in diagnosing early pregnancy diagnosis as early as 25 days post breeding.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.10
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• pp.1404-1410
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• 2008

This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

• Korean Journal of Veterinary Service
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• v.25 no.2
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• pp.117-126
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• 2002

Contamination levels of pathogenic microorganisms in 145 cases of beef, which were distributed in Gwangju province, had been investigated in each distributed stage and also monitored by general bacterial count and E coli count index. General bacterial count of beef from the slaughterhouse was 10$^4$cfu/g less than the level of promotion(10 cfu/$\textrm{cm}^2$) and E coli count index was also under the level of 10$^2$cfu/$\textrm{cm}^2$ recommended level of the ministry of agriculture and forestry. Pathogenic microorganisms were detected from 23.2% of samples in the consumption stage, 12.5% in the slaughtering stage and 5.6% in the transporting and processing stage. Staphylococcus aureus was isolated in the largest number and its ratio was 9.0%, listeria monocytogenes 5.5% and salmonella spp 1.4%. There were no samples that bacteria had been detected dually. E coli O157:H7 and campylobacter jejuni were not isolated. In raw and chilled beef, isolation rate of pathogenic microorganisms were 13.3% and 16.5% each. Especially in raw beef, L monocytogenes was. isolated in 3 samples among 30 cases (10%) and S aureus in one sample (3.3%). According to a scale of meat store, isolation rates of pathogenic microorganisms were different. It was 28.6% in the small-scale meat store and 16.7% in the large-scale meat store each. Four cases (16.7%) of S aureus were isolated in the large-scale meat store and seven cases (20.0%) of L monocytogenes and 2 cases (5.7%) of salmonella spp were isolated in the small-scale meat store. S aureus was isolated in two places among 10 feeding facilities of the elementary school. This result shows that the sanitation of elementary school feeding facilities is so poor and more careful policy consideration is needed. Eleven strains of S aureus isolated showed ${\beta}$-hemolysis on blood agar, 1 strain ${\alpha}$-hemolysis, and 1 strain ${\gamma}$-hemolysis. Isolated strains of L monocytogenes were reconfirmed in 560 bp by PCR. Conclusively, these results show that the sanitary condition in the stages of slaughtering, transportation-processing and consumption influences the degree of pathogenic microorganisms contamination in beef severely It is necessary to apply thoroughly hazard analysis critical control point in a process of beef distribution and also to develop rapid test methods for microorganism diagnosis. This effort is very important for the supply of safe and clean meat from farm to table and helpful for the improvement of public health.

• Korean Journal of Agricultural Science
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• v.42 no.1
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• pp.29-35
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• 2015

This study was performed to analyze the distribution and antimicrobial resistance of pathogenic microorganisms isolated from the carcass and environments of chicken processing plant located in Gyeonggi province from October to November in 2010. Chicken slaughterhouse was visited 3 times and totally 40 samples were collected from chicken carcass before and after washing (n=14), chicken cuts (n=7), cooling water (n=8), brine (n=2), cutting knives (n=7) and working plate (n=2). Whole-chicken rinsing technique (for chicken carcasses) and swab technique (for working plate and knives) were used to analyze the distribution of pathogenic microorganisms. In addition, brine and chilling water from storage tanks were gathered using sterilized tubes and used as samples. The matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for whole cell fingerprinting in combination with a dedicated bioinformatic software tool was used to identify the isolated microorganisms. The pathogenic microorganisms, such as Bacillus cereus (n=8) and Staphylococcus aureus (n=9), were isolated form the chicken processing process (chicken carcasses of before and after chilling, chicken cuts, and working plate). The antimicrobial susceptibility of those isolated microorganisms was analyzed using 21 antimicrobial agents. In the case of B. cereus, it showed 100% of resistance to subclasses of penicillins and peptides, and it also resistant to cephalothin, a member of critically important antimicrobials (CIA), however there was no resistance (100% susceptible) to vancomycin and chloramphenicol. S. aureus showed 100% resistance to subclasses of peptides and some of penicillins (penicillin and oxacillin), however, it showed 100% susceptibility to cephalosporins (cefazolin and cephalothin). All of the tested pathogens showed multi drug resistance (MDR) more than 4 subclasses and one of B. cereus and S. aureus showed resistance to 9 subclasses. After the ban on using the antimicrobials in animal feed in July 2011, there would be some change in microbial distribution and antimicrobial resistance, and it still has a need to be analyzed.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.69-69
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• 2002

Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1695-1701
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• 2002

The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.10
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• pp.1656-1665
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• 2020

Objective: This study aimed to determine the effects of stress during slaughter of beef cattle on physiological parameters, carcass, and meat quality at a Federal Inspection Type slaughterhouse located in the southeast of Mexico. Methods: A total of 448 carcasses of male Zebu×European steers with an average age of 36 months were included. Carcass assessment of presence of bruises and bruise characteristics was carried out on each half-carcass. Blood variable indicators of stress (packed cell volume, neutrophil to lymphocyte ratio, glucose, cortisol concentration) and meat quality parameters (pH, color, shear force, drip loss) were evaluated. Results: Of the 448 carcasses evaluated, 81% of the carcasses showed at least one bruise; one bruise was detected in 36.6% and two bruises in 27.0% of animals. Of the 775 bruises found, 69.2% of the bruises were grade 1 in region 3. Of the 448 carcasses studied, 69.6% showed hyperglycemia (6.91 mmol/L); 44.3% and 22.7% showed high (74.7 ng/mL) and extremely high (108.8 ng/mL) cortisol levels, respectively, indicative of inadequate handling of animals during preslaughter and slaughter. Of the carcasses evaluated, 90.4% had a pH ≥5.8 with an average of pH 6.3. In both pH groups, meat samples showed L^⋆ values >37.0 (81.6%) and a shear force >54.3 N; meat pH≥5.8 group showed a drip loss of 2.5%. These findings were indicative of dark, firm, and dry (DFD) meat. According to principal component analysis, grades 1 and 2 bruises in region 3 and grade 1 bruises in region 5 were highly associated with cortisol, drip loss, and color parameters b^⋆ and h^⋆ and were negatively associated with L^⋆, a^⋆, and C^⋆. Conclusion: The bruises probably caused by stress-inducing situations triggered DFD meat. Appropriate changes in handling routines in operating conditions should be made to minimize stress to animals during the slaughter process to improve animal welfare and meat quality.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.987-997
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• 2016

The objective of the study was to compare the carcass characteristics, meat quality, and blood parameters of slow and fast grown female broiler chickens fed in organic or conventional production system. The two genotypes tested were medium slow-growing chickens (SG, Hubbard Red JA) and commercial fast-growing chickens (FG, Ross 308). Both genotypes (each represented by 400 chickens) were divided into two sub-groups fed either organic (O) or conventional (C) systems. Chickens of each genotype and system were raised in a semi environmentally controlled poultry house until 21 d of age and were assigned to 5 pens of 40 chickens each. Then, O system chickens were transferred into an open-side poultry house with an outdoor run. At 81 d of age, 10 female chickens from each genotype and from each production system (n = 40) were randomly chosen to provide material for analysis, and were weighed and brought to the slaughterhouse to assess carcass characteristics and meat quality. The blood parameters were determined by using 5 female chickens from each genotype and from each production system (n = 20). FG had the higher live weight, along with carcass, breast, and thigh-drumstick weights compared to SG (p<0.05). FG had the higher breast yield, whereas SG had the higher thigh-drumstick yield (p<0.05). The O system resulted in a higher amount of abdominal fat (p<0.05). In addition, the O system values were higher for dry matter, crude ash, crude protein, and $pH^{15}$ values in breast meat, and for crude ash, crude protein, and $pH^{15}$ values in drumstick meat (p<0.05). In addition, total saturated fatty acids, total mono-unsaturated fatty acids, and total omega 3 were significantly higher in the O system than in the C system. Thus, the O system showed a positive advantage compared to the C system regarding female chicken meat quality, primarily within the ash, protein, and total omega 3 fatty acid profiles. In conclusion, the present study indicated that the main factor affecting the carcass characteristics of female chickens was genotype, whereas the organic system contributed to enhanced meat quality. These findings provide a better understanding of the relative roles of genotype and production systems in female broiler characteristics, and might aid producers in designing their facilities to optimize yield and quality while maintaining acceptable animal welfare standards.