We have developed a novel polymerase chain reaction (PCR)-microchip gel electrophoresis (MGE) method based on the sieving gel mixture of commercially available poly(vinylpyrrolidone) (PVP) and hydroxy ethyl cellulose (HEC) for the rapid detection and diagnosis of the orf virus (ORFV) from Korean indigenous goat. After amplification of 594-bp DNA fragment from the B2L gene of ORF virus, the amplicon was analyzed by the MGE separation. The glass microfluidic chip (64 mm total length (36 mm effective length)${\times}$90 ${\mu}$m width${\times}$20 ${\mu}$m depth) allowed the fast detection and diagnosis of ORFV in the mixture of 1.0% PVP ($M_r$ 360,000) and 1.0% HEC ($M_r$250,000) as a sieving matrix with better resolution and reproducibility of DNA fragments. Under the electric field of 277.8 V/cm, the 594-bp DNA was analyzed within 4 min. Compared to traditional slab gel electrophoresis, the PCR-MGE method was twenty times faster and an effective clinical method for the quantitative analysis of ORFV.
Hong, Sung Wook;Bae, Hyo Ju;Chang, Jin Hee;Kim, So-Young;Choi, Eun-Young;Park, Beom Young;Chung, Kun Sub;Oh, Mi-Hwa
Journal of Dairy Science and Biotechnology
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v.31
no.2
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pp.153-159
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2013
Lactic acid bacteria are microorganisms that are closely associated with human and/or animal environments, and are categorized as generally recognized as safe (GRAS) organisms due to their ubiquitous appearance in foods and their contribution to the healthy microflora of mucosal surfaces. This study was performed to isolate and identify lactic acid bacteria with antagonistic effects against food-borne pathogens. A total of 3,000 acid-producing bacteria were isolated from infant feces, cattle feces, goat feces, dog feces, pig feces, vaginal tracts, vegetables, fruits, Kimchi, Jeotgal, fermented sausages, raw milk, cheese, yogurt, Cheonggukjang, Meju, and Makgeolli cultured on MRS agar with 0.05% bromocresol purple. For the isolation of bacteriocin-producing bacteria, the diameter of the clear zone was measured on MRS agar plates. Twenty-six isolates exhibited strong antibacterial activity against indicator strains such as Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis. Lactic acid bacteria were identified as Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus curvatus, Lactobacillus plantarum, and Pediococcus acidilactici by 16S rDNA gene sequence analysis. The results of this study suggest that the isolates could be used as potential probiotic starters for functional food applications.
This trial was carried out to determine effects of organic feeds in comparison to conventional diet on feed intake, digestibility, and nitrogen retention in Korean native goats. Twelve Korean native goats were allotted to treatments in four groups of three goats and then they were housed in separate metabolism cages for 21 days. Treatments included conventional diet (A) as a control group and three organic feed groups (B: organic rice straw, C: organic nee leaves, D: organic mixture of rice straw and tree leaves). The A treatment, conventional diet, consisted of common rice straw and commercial concentrates at a proportion of 60 and 40%, respectively. All ingredients of organic feeds treatments were organically produced-agricultural products without any application of chemical fertilizer and pesticide. Four experimental diets were formulated to have the same ratio of forage to concentrate and similar contents for protein and carbohydrate across treatments and they were offered to goats ad libitum. Feed intake, apparent nutrient digestibility and nitrogen retention were investigated. For chemical compositions of experimental diets, all nutrients except crude ash and ether extract were not significantly different across treatments as we expected. Crude ash content was highest in the A treatment (P<0.05), however, it was not significantly different among organic feeds treatments. Ether extract content was higher (P<0.05) in C and D treatments than in A and B. Even if dry matter intakes for organic feeds treatments were not significantly different among them, they were significantly higher (P<0.05) compared with conventional diet. Fecal excreta were not significantly different across treatments, resulting in significantly higher digestible dry matter (g/day) in treatments of organic feeds (P<0.01). Average daily gain (ADG) and feed efficiency (FE) were more increased (P<0.01) in treatments of organic feeds compared with conventional diet. Digestibilities for most of nutrients except NFC had the same trend as ADG and FE, however, NFC digestibilities for C and D treatments were significantly lower (P<0.01) than those of A and B. Nitrogen intakes for organic feeds treatments were significantly higher (P<0.001) than conventional diet, with no difference among organic feeds treatments. Fecal nitrogen loss was higher (P<0.05) for C and D treatments than for A and B. Retained nitrogen contents were significantly higher (P<0.05) for organic feeds treatments than for conventional diet, but nitrogen retention rate did not show any difference across treatments. The results showed that organic feed supplementation more improved feed intake, digestibility and nitrogen retention in comparison with conventional diet, and thus they could be concluded that organic feeds might contribute to animal performance and a safer production of animal product.
To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.
Park, Jong-Keun;Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
Food Science of Animal Resources
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v.27
no.2
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pp.209-215
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2007
In order to develop a sensitive and reliable method for the species-specific molecular markers, PCR-RFLP assay of the mitochondrial DNA(mt DNA) 12S rRNA gene was exploited for the identification of the origin of animal meat species including cattle, pig, sheep, goat, horse, deer, chicken, duck and turkey. A specific primer pairs were designed, based on the nucleotide sequences of mt 12S rRNA gene, for the amplification of the highly conserved region in the gene of the animal species using PCR-RFLP technique. mt DNA was isolated from meat samples followed by DNA amplification using PCR with the specific primers. PCR amplification produced an approximately 455 bp fragment in each of these animal meats. To distinguish pleat species, the PCR amplicons were digested with restriction endonucleases Tsp5091 and MboI, respectively, which generates distinct RFLP profiles. The DNA profiles digested with Tsp5091 allowed the clear discrimination in the mammalian meat species and the DNA profiles digested with MboI in poultry meat species. Therefore, the PCR-RFLP profiles of mt 12S rRNA gene could be very useful to identify the origin of the raw materials in the raw meats as well as the processed meat products.
The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.
Pang, X.S.;Wang, Z.Y.;Zhu, T.G.;Yin, D.Z.;Zhang, Y.L.;Meng, L.;Wang, F.
Asian-Australasian Journal of Animal Sciences
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v.23
no.2
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pp.188-196
/
2010
The objective of this study was to characterize the litter sizes in Huanghuai goats with high prolificacy (HP, five or more kids born per litter on at least one occasion), and to compare their peripheral blood concentrations of progesterone and estradiol with those of goats with poor prolificacy (PP, up to three kids born per litter on any occasion). The circulating concentrations of progesterone and estradiol were measured by radioimmunoassay from daily blood samples taken during natural estrus cycles and at 1-5 days after ovariectomy. Estrus was synchronized using two doses of a prostaglandin analog. Litter size for the HP goats increased up to a parity of five and decreased thereafter. The percentage of goats with litter sizes of $\geq$4 from parities 3 to 6 ranged from 44.5% to 58.3%. Although small differences in litter size were obtained for goats of parities three, four and six relative to five, parity five does had the highest mean litter size. Progesterone concentrations began to rise earlier and were higher in the HP than in the PP goats on most days of the luteal phase, but not during the follicular phase of the cycle or after ovariectomy. There was a significant difference between the two groups (p<0.05) in the magnitude of the progesterone plateau. Mean estradiol concentrations in the HP group remained significantly higher than in the PP group (p<0.05) during the estrus cycle. There were two estradiol peaks in the HP goats during the early luteal phase, but only one in the PP group. Measurements of individual corpora lutea (CL) in vitro showed that there was a greater prevalence of small CL (<6 mm in diameter) in the HP group than in the PP group (p<0.05). After ovariectomy, the estradiol level on day 1 was significantly higher than at the nadir during the estrus cycle in both the HP (p<0.01) and PP (p<0.05) goats, while levels decreased by 12.3% and 26.2% respectively compared with the mid-luteal period in HP and PP goats (p<0.05). The overall mean estradiol concentrations in HP goats were lower than in the PP group, but no significant differences were found between groups at 1-5 days after ovariectomy.
This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.
The effect of high and low level of feed intakes on nutrient digestibility, nutrient losses through methane, energy and protein utilization by goats fed on alfalfa (Medicago sativa L.) pellets based diets was investigated in this study. Twelve castrated Japanese goats were employed in two subsequent digestion and metabolism trials. The goats were divided into three groups, offered three diets. Diet 1 consisted of 100% alfalfa pellet, Diet 2 was 70% alfalfa pellet and 30% corn, and Diet 3 was 40% alfalfa pellet and 60% corn. The two intake levels were high (1.6 times) and low (0.9 times) the maintenance requirement of total digestible nutrients (TON). Rumen ammonia nitrogen ($NH_3$-N) level of Diet 1 was lower (p<0.001) compared to Diets 2 and 3, but the values were always above the critical level (I50 mg/liter), The pH values of rumen liquor ranged from 6.02 to 7.30. Apparent digestibility of nutrient components did not show differences (p>0.05) between the two intake levels but inclusion of corn significantly altered the nutrient digestibility. Diet 3 had highest (p<0.001) dry matter (DM), organic matter (OM), ether extract (EE) and nitrogen fee extract (NFE) digestibility followed by the Diet 2 and Diet 1. The crude protein (CP) digestibility values among the three diets were in a narrow range (70.1 to 70.8%). Crude fiber (CF) digestibility for Diet 3 was slight higher (p>0.05) than that for other two diets. When alfalfa was replaced by corn, there were highly significant (p<0.001) increases in DM, OM, EE and NFE apparent digestibility and a slight increase in the CF digestibility (p>0.05). There were no differences (p>0.05) in energy losses as methane ($CH_4$) and heat production among the diets but energy loss through urine was higher for the Diet 1. The total energy loss as $CH_4$ and heat production were higher for the high intake level but the energy loss as $CH_4$ per gram DM intake were same (0.305 kcal/g) between the high and low intake level. Retained energy (RE) was higher for Diet 3 and Diet 2. Nitrogen (N) losses through feces and urine were higher (p<0.001) for Diet 1. Consequently, N retention was lower (p>0.05) for Diet 1 and higher in Diets 3 and 2. It is concluded that inclusion of corn with alfalfa increased the metabolizable energy (ME) and RE, and retained N through reducing the energy and N losses. The high level of intake reduced the rate of nutrient losses through feces and urine.
A comparative study on the quality of meat from three breeds of goats reared in Bulgaria: Local Aboriginal (LA); Bulgarian White Dairy (BWD) and crossbreeds of local aboriginal with Saanen goats (LA ${\times}$ S). Eight intact young male goats from each breed have been included in the experiment. The animals from the three breeds have been reared under similar conditions. The animals have been slaughtered at the age of 2 months (at weaning) and at the age of 6 months (upon reaching sexual maturity). The following tests have been performed on samples of m. longissimus dorsi between $5^{th}$ and $6^{th}$ ribs: pH 45 min post mortum, colorimetrically color of meat, water holding capacity and fatty acid composition of the fat. The thickness of muscle fibre and the proportion of muscle, connective and adipose tissue have been microscopically determined at m. longissimus dorsi, m. semitendinosus and m. iliopsoas. After deboning and separating the bigger tendons, the chemical composition of the meat from the left half of the carcass has been determined. The results reveal lack of statistically significant differences among the three breeds concerning pH, the water holding capacity, color of meat, thickness of the muscle fibre and fatty acid composition of the fat. The carcass meat of crossbreeds of LA ${\times}$ S contains significantly more fat than the other two breeds. This influences the proportion of muscle, connective and adipose tissue in m. longissimus dorsi, m. semitendinosus and m. iliopsoas, resulting in more adipose tissue in the LA ${\times}$ S compared to young goats of the LA or the BWD. There are differences in slaughtering 2-month-old goats (at weaning) and 6-month-old ones (upon reaching sexual maturity). At a greater age the content of fat in the carcass increases, as well as the quantity of the adipose tissue, the intensity of the colour of the meat and there is a tendency towards thickening of the muscle fibre and increasing the water holding capacity of meat. The moisture content in meat decreases due to the increase of fat.
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