• Asian-Australasian Journal of Animal Sciences
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• 제15권10호
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• pp.1412-1421
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• 2002

To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1374-1388
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• 2007

Three experiments were conducted to investigate the hypothesis that cows grazing on pasture produce the highest proportion of c-9 t-11 CLA in milk fat and no further increase can be achieved through supplementation of diets rich in linoleic acid, such as full-fat extruded soybeans or soybean oil. In experiment 1, 18 lactating Holstein cows were used in a randomized complete block design with measurements made from wk 4 to 6 of the experiment. In experiment 2, three cannulated lactating Holstein cows were used in a $3{\times}3$ Latin square design. Each period was 4 wk with measurements made in the final wk of each period. Cows in both experiments were assigned at random to treatments: a, conventional total mixed ration (TMR); b, pasture (PS); or c, PS supplemented with 2.5 kg/cow per day of full-fat extruded soybeans (PES). In both experiments, feed intake, milk yield, milk composition, and fatty acid profile of milk and blood serum were measured, along with fatty acid composition of bacteria harvested from rumen digesta in experiment 2. In experiment 3, 10 cows which had continuously grazed a pasture for six weeks were assigned to two groups, with one group (n = 5) on pasture diet alone (PS) and the other group (n = 5) supplemented with 452 g of soy oil/cow per day for 7 d (OIL). In experiment 1, cows in PS treatment produced 350% more c-9, t-11 CLA compared with cows in TMR treatment (1.70 vs. 0.5% of fat), with no further increase for cows in PES treatment (1.50% of fat). Serum c-9, t-11 CLA increased by 233% in PS treatment compared with TMR treatment (0.21 vs. 0.09% of fat) with no further increase for cows in PES treatment (0.18% of fat). In experiment 2, cows in PS treatment produced 300% more c-9 t-11 CLA in their milk fat compared with cows in TMR treatment (1.77 vs. 0.59% of fat), but no further increase for cows in PES treatment (1.84% of fat) was observed. Serum c-9, t-11 CLA increased by 250% for cows in PS treatment compared with cows in TMR treatment (0.27 vs. 0.11% of fat), with no further increase for cows in PES treatment (0.31% of fat). The c-9, t-11 CLA content of ruminal bacteria for cows in PS treatment was 200% or more of TMR treatment, but no further increase in bacterial c-9, t-11 CLA for cows in PES treatment was observed. Supplementation of soy oil in experiment 3 also did not increase the c-9 t-11 CLA content of milk fat compared with cows fed a full pasture diet (1.60 vs. 1.54% of fat). Based on these findings, it was concluded that supplementing with feeds rich in linoleic acid, such as full-fat extruded soybeans or an equivalent amount of soy oil, to cows grazing perennial ryegrass pasture may not increase milk fat c-9 t-11 CLA contents.

• 한국방사선학회논문지
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• 제5권6호
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• pp.391-400
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• 2011

폐렴은 호흡기계의 감염이고 원인균, 병인, 침범부위, 그 밖의 여러 가지 상황에 따라서 다양하게 분류된다. 비정형 병원균주 페렴으로 의심되어 내원한 46세의 남자 환자에서 이학학적 소견이나 혈액검사, 객담도말검사, 소변검사, 기생충검사, 기관지내시경검사, 침생검 등에서 특이할만한 원인균을 찾지 못했으며, 청진이상, 고열, 고혈압, 객담, 호흡곤란 등의 증상 또한 보이지 않았다. 세균성 또는 비정형 병원균의 광범위치료 항생제 복용이나 기생충제제를 복용하였으나 재발되었으며, 자연치유 및 재발이 반복되며 호전되었다. 반흔을 남기며 호전되고 새로운 부위에 결절이 재발하기를 반복하면서 서서히 없어지는 기간은 평균 20일 정도였다. 재발 이후 흉부엑스선 촬영과 흉부 고해상 전산화단층촬영을 추적 검사한 결과 흉부엑스선 촬영에서는 특이한 징후를 관찰하지 못했으나 고해상 전산화단층촬영에서는 병변이 호전되어가고 새로운 부위에 재발되는 모습을 관찰할 수가 있었다. 양측 하부 폐에 재발성 경과를 취한 비정형병원균주 폐렴이 의심되는 환자의 경우, 흉부엑스선 촬영 소견은 횡격막이나 간(Liver), 척추 등에 의해 숨기 때문에 추적검사로서 도움을 주는 데는 한계가 있으며, 흉부 고해상 전산화단층촬영 검사를 하여 비교하는 것이 바람직하다. 저자는 재발성 경과를 취한 비정형 병원균주 폐렴1예에 대한 문헌고찰과 함께 보고한다.

• 한국식품영양과학회지
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• 제43권5호
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• pp.756-762
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• 2014

전통 장류 유래 효소 활성 및 항균 활성 우수 Bacillus sp.를 스타터로 한 콩 발효물의 품질 특성을 분석하였다. 총균수는 HJ5-2를 접종한 시료가 $3.00{\times}10^9CFU/mL$로 가장 높았으며, 유산균은 스타터 첨가 콩 발효물에서 $2.50{\times}10^2{\sim}7.30{\times}10^4CFU/mL$를 나타나는 것으로 보아 접종된 균주가 우점종이 된 것을 확인하였다. ${\alpha}$-Amylase 활성은 RD7-7(20.82 unit/mL)과 HJ18-4(18.45 unit/mL), protease 활성은 RD7-7(647.92 unit/mL)을 접종한 콩 발효물이 높은 활성을 나타내었다. 환원당 함량은 HJ18-4(2.35%), 아미노태 질소 함량은 RD7-7(227.96 mg%) 접종 콩 발효물에서 높은 함량을 나타내었다. 아미노산 함량은 glutamic acid, aspartic acid, leucine, lysine, arginine 순으로 검출되었다. 유기산은 oxalic acid(36.51~63.57 mg/100 g)가 모든 실험군에서 검출되었으며, succinic acid(600.15~429.49 mg/100 g)는 control과 RD7-7 접종 콩 발효물에서 검출되었다. 향기 성분을 분석한 결과 총 28종(esters 3종, ketones 6종, aldehydes 2종, alcohols 4종, hydrocarbons 2종, pyrazines 2종, acids 4종, 기타 4종)이 검출되었으며, RD7-7과 HJ18-4를 접종한 콩 발효물에서 23종의 향기 성분이 검출되었다. 본 연구는 고품질 장류용 스타터 개발을 위한 단일균 접종 콩 적용 시험으로 향후 혼합배양 스타터 개발 및 기능성 콩 발효소재 개발을 위한 기초자료로 제시하고자 한다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Asian-Australasian Journal of Animal Sciences
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• 제14권4호
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• pp.479-484
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• 2001

The effect of dietary supplementation of sodium salt of isobutyric acid in low protein (10% CP) wheat straw based diet on nutrient utilization and rumen fermentation was studied in ruminally fistulated male crossbred cattle. The study included a 7 day metabolism and a 3 day rumen fermentation trials. The cattle were distributed into two equal groups of 4 each. The animals of control group were fed a basal diet consisting of wheat straw, concentrate mixture and green maize fodder in 40:40:20 proportion whereas branched chain volatile fatty acid (BCFA) supplemented group received a basal diet + isobutyric acid at 0.75 percent of basal diet. The duration of study was 36 days. The feed intake between experimental groups did not differ significantly and the average total DMI (% BW) was 2.01 and $2.28kg\;day^{-1}$ in control and BCFA supplemented diets. The dietary supplementation of BCFA improved (p<0.05) the DM, OM, NDF and cellulose digestibility by 4.46, 6.63, 10.57 and 11.31 per cent over those fed control diet. The total N retention on BCFA supplementation was improved (p<0.01) due to decreased (p<0.05) urinary N excretion. The concentrations of ruminal total N was 37.07 and $34.77mg\;100ml^{-1}$ in control and BCFA fed groups, respectively. Dietary supplementation BCFA significantly (p<0.01) reduced the ruminal ammonia N concentration as compared to control and the mean values ($mg\;100ml^{-1}$) were 13.18 and 9.42 in control and BCFA fed groups. The total VFA concentration was higher (p<0.01) in BCFA supplemented group (101.14 mM) than the control (93.05 mM). Among the VFAs, the molar proportion of acetate was higher (p<0.01) in BCFA supplemented group (71.07 mM) as compared to control (64.98 mM). However, the concentration of propionate and butyrate remained unchanged. Amino acids composition of bacterial hydrolysates was similar in both the groups. Ruminal outflow rate of liquid digesta was higher (p<0.01) in BCFA fed group ($67.56l\;day^{-1}$) than control ($52.73l\;day^{-1}$). It is concluded that the dietary supplementation of Na-salt of isobutyric acid in low protein diet improved the nutrient utilization and ruminal fermentation characteristics.

• Asian-Australasian Journal of Animal Sciences
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• 제20권4호
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• pp.569-576
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• 2007

Four hundred and fifty tilapias ($6.77{\pm}0.23$ g) were assigned randomly to six groups to evaluate the feasibility of the tested antibacterial peptides (ABPs) and oligosaccharides as substitutes for antibiotics. The control group was fed with a commercial tilapia diet; other five groups were fed with the same commercial diet supplemented with konjac glucomannan (KGLM), cluster bean galactomannan (CBGAM), and three animal intestinal ABPs derived from chicken, pig and rabbit at 100 mg/kg respectively. After 21 days of feeding, growth, disease resistance, and in vivo anti-adherence were determined. Furthermore, the inhibitory effect of tested agents on adhesion of Aeromonas veronii biovar sobria (A.vbs) strain BJCP-5 to tilapia enteric epithelia in vitro was assessed by cell-ELISA system. As a result, the tested agents supplemented at 100 mg/kg show significant benefit to tilapia growth and disease resistance (p<0.05), and the benefit may be correlated with their interfering in the contact of bacteria with host mucosal surface. Although none of the tested agents did inhibit the growth of BJCP-5 in tryptic soy broth at $100{\mu}g/ml$, all of them did inhibit the adhesion of A.vbs to tilapia enteric epithelia in vivo and in vitro. In vitro mimic assays show that three ABPs at low concentrations of $25{\mu}g/ml$ and $2.5{\mu}g/ml$ have the reciprocal dose-dependent anti-adherence effect. The inhibition of ABPs may be correlated with a cation bridging and/or receptor-ligand binding, but not with hydrophobicity. The KGLM and CBGAM inhibited the adherence of BJCP-5 to tilapia enteric epithelia with dose-dependent manner in vitro, and this may be through altering bacterial hydrophobicity and interfering with receptor-ligand binding. Our results indicate that the anti-adherence of the tested ABPs and oligosaccharides may be one of the mechanisms in promoting tilapia growth and resistance to A.vbs.

• Asian-Australasian Journal of Animal Sciences
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• 제25권8호
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• pp.1124-1131
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• 2012

Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.

• Asian-Australasian Journal of Animal Sciences
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• 제26권6호
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• pp.788-794
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• 2013

Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. ${\alpha}_1$-anti trypsin, ${\alpha}_1$-acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with $SCC{\geq}2{\times}10^5$ cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that ${\alpha}_1$-anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of ${\alpha}_1$-acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC{\geq}2{\times}10^5$ cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that ${\alpha}_1$-anti trypsin indicates SCM irrespective of etiology, whereas ${\alpha}_1$-acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good indicator of SCM. It is recommended measuring both ${\alpha}_1$-anti trypsin and ${\alpha}_1$-acid glycoprotein in milk to diagnose SCM in buffalo irrespective of etiology.

• 한국버섯학회지
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• 제18권3호
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• pp.208-213
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• 2020

양송이배지로부터 분리한 인산가용화균 Burkholderia contaminans PSB-A와 Burkholderia ambifaria PSB-B의 단일접종 및 동시접종에 따른 인산가용화 능력의 시너지 효과 및 상추생육효과를 조사하였다. 인산캄슘이 함유된 NBRIP 액체배지에 분리균을 접종하고 해리된 가용인산(soluble phosphorus)함량을 HPLC에 의해 분석하여 인산가용화능을 측정한 결과, 배양 3일차에 동시접종구 166.3 ㎍ mL^-1, B. contaminans 143.7 ㎍ mL^-1, B. ambifaria 127.1 ㎍ mL^-1의 순으로 나타나 두 종의 인산가용화세균간의 동시접종에 의한 시너지효과가 다소 관찰되었다. 배지 내 pH 와 잔류 glucose 함량 변화도 모든 접종구에서 배양 1 - 3일 후 pH 7.0에서 pH 4.0 수준으로 감소하였으며, glucose 함량은 초기 10 mg mL^-1 에서 4.3 mg mL^-1 수준으로 검출되어 인산가용화능과 거의 유사한 경향을 보였다. 배양여액의 유기산 분석결과 gluconic acid 약 12.3 mg mL^-1 와 malic acid 약 7.9 mg mL^-1 수준으로 가장 높게 검출되었으며, 결과적으로 두 세균이 분비하는 유기산이 배지의 pH를 감소시키고, 인산 가용화를 유도하는 주요 원인임을 확인할 수 있었다. 접종 4주 후 상추 생육차이를 조사한 결과, TCP를 첨가하지 않은 B. contaminans 접종구, B. ambifaria 접종구 와 동시 접종구에 비해 TCA 첨가한 모든 처리구에서 약 7-9% 수준의 생육증진효과가 보였으나, 단일접종과 동시접종구간에 유의수준의 차이를 확인할 수 없었다.