• Asian-Australasian Journal of Animal Sciences
• /
• 제20권11호
• /
• pp.1721-1728
• /
• 2007

An in vitro and a feeding trial were conducted to investigate the effect of xylanase supplementation on the feeding value of growing pig diets containing high proportions of Chinese double-low rapeseed meals (DLRM). Seven diets were formulated to meet NRC (1998) nutrient requirements. Diet 1 based on corn-soybean meal was used as positive control 1, and diet 2, a practical diet which incorporated a conventional level of Chinese DLRM (60 g/kg diet), as positive control 2. Diet 3 contained a higher level of DLRM (100 g/kg diet) as the negative control. Diet 3 plus xylanase at 0.10, 0.25, 0.50 and 0.70 g/kg diet created diets 4, 5, 6 and 7, respectively. The seven diets were incubated in triplicate with the in vitro two-stage enzyme incubation method to predict responses of diets to xylanase in terms of digestibility of dry matter (DM), crude protein (CP) and neutral detergent fibre (NDF). In vitro, the negative control had the lowest CP and NDF digestibility. Both DM and CP digestibility were increased (p<0.05) owing to xylanase supplementation either at 0.50 or 0.70 g/kg diet, and NDF digestibility was improved following xylanase addition at all of the test levels. There was a high linear correlation ($r^2>90$, p<0.05) between the activity concentration of the enzyme when transformed into its logarithmic value and in vitro digestibility coefficients of DM, CP or NDF. In the feeding trial, 112 crossbred pigs were randomly assigned to seven dietary treatments with 16 replicate pens of one pig each. An obvious dose effect on growth rate was observed ($r^2=0.79$, p<0.05) within the inclusion levels of xylanase. Compared with the negative control, xylanase addition at 0.70 g/kg diet resulted in significantly increased ADG (878 g/d vs. 828 g/d, p<0.05), and a tendency towards improved growth rate (868 g/d vs. 828 g/d, p = 0.10) was also observed following the inclusion of xylanase at 0.50 g/kg diet. It would appear that the nutrient utilization of corn and Chinese DLRM diets by pigs could be enhanced by an appropriate amount of xylanase addition. The in vitro and in vivo results suggested that the in vitro incubation method is feasible for predicting responses of pigs to exogenous enzymes and identifying those preparations that possess potential for improvement of the nutritive values of feedstuffs.

• Applied Biological Chemistry
• /
• 제41권1호
• /
• pp.47-52
• /
• 1998

3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) 환원효소는 식물의 병원균 방어물질 (phytoalexin), 광합성 색소 (the phytol of chlorophyll), 성장 호르몬 (abscisic acid와 gibberellin 등) 및 스테롤 (phytosterol) 등의 생합성에 관여하는 주효소이다. 암조건 하에서 발아 후 4일째의 벼 유묘 microsome을 재료로 비이온성 detergent 인 Brij W-1 (final 0.4%)을 사용하여 가용화 시킨 후, DEAE-Sephadex A-50 크로마토그래피 칼럼과 hexaneagarose를 matrix로 기질인 HMG-CoA를 결합시켜 제조한 친화성 크로마토그래피 칼럼을 이용하여 세포막 결합효소인 HMG-CoA 환원효소를 정제하였다. 정제된 HMG-CoA 환원효소의 최종 회수율은 7.14% 였고, 분자량은 10% SDS-PAGE에서 55 kDa이였다 HMG-CoA 환원효소의 최적 반응 온도는 $37^{\circ}C$, 최적 반응 pH는 6.9였고, 기질인 HMG- CoA에 대한 HMG-CoA 환원효소의 $K_m$ 및 $V_{max}$ 값은 $180\;{\mu}M$과 107 pmol/mg, 수소공여체인 NADPH에 대한 $K_m$과 $V_{max}$값은 $810\;{\mu}M$과 32.1 pmol/min/mg이였다.

• 대한수의학회지
• /
• 제37권3호
• /
• pp.583-593
• /
• 1997

Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

• Journal of Animal Science and Technology
• /
• 제44권5호
• /
• pp.549-560
• /
• 2002

본 시험은 패스틴$^{(R)}$ 을 첨가하였을 때, in vitro 상에서 단백질 fraction과 분해율에 미치는 영향과, in vivo 상에서 반추위 성상, 미생물 군집, 암모니아태 질소 농도 및 영양소 소화율에 미치는 영향을 구명하고자 실시하였다. In vitro 실험에서는 1mm로 분쇄된 대두박을 기질로 하여 패스틴$^{(R)}$ ((주)은진인터내셔날)을 첨가하여 borate-phosphate buffer와 중성세제에서의 조단백질 분해율을 측정하였으며, exogenous enzyme (Streptomyces griseus 유래 protease)를 이용하여 39$^{\circ}C$에서 0, 2, 4, 8, 12, 48 시간동안 배양 후 조단백질 분해율을 측정하였다. 반추위 발효성상과 영양소 소화율은 반추위 fistula가 부착된 평균체중 300kg의 홀스타인 수소 4두를 이용하여 무첨가구, 패스틴$^{(R)}$ 첨가구의 두개 처리구에 2마리씩 4마리를 배치하여 측정하였다. Buffer Soluble Protein fraction은 패스틴$^{(R)}$ 첨가 수준별로 차이가 없었으나, 무첨가구에 비해 패스틴$^{(R)}$ 첨가구에서 감소하는 경향을 보였다. 단백질 분해율은 배양 0 시간대에서 4시간대까지는 처리구간 유의성이 없었지만, 12 h과 48 h에서는 패스틴$^{(R)}$ 첨가로 시험구에서 감소되었다. 용해 단백질 분해율 ‘a’는 패스틴$^{(R)}$ 시험구에서 경미하게 높은 수치를 나타내었지만, 소화 가능한 단백질 분해율 ‘a+b’는 패스틴$^{(R)}$ 시험구에서 낮은 경향을 보였다. 패스틴$^{(R)}$ 첨가로 pH와 $NH_3$- N 농도는 증가하는 경향이었으며 휘발성지방산, 미생물 수 및 enzyme activity는 감소하였고 영양소 소화율은 높았으나 유의적인 차이는 없었다.

• 한국미생물·생명공학회지
• /
• 제44권3호
• /
• pp.333-340
• /
• 2016

호알칼리성 protease를 분비하는 토양에서 분리된 Bacillus sp. DK1122 균주로부터 효소의 생산조건을 검토 후, 효소를 정제하고 특성을 알아보았다. 본 균주의 효소생산 최적 배지조성은 0.5% (w/v) glucose, 0.8% (w/v) yeast extract, 0.5% (w/v) polypeptone, 0.1% (w/v) K₂HPO₄, 0.02% (w/v) MgSO₄· 7H₂O, 1% (w/v) Na₂CO₃, 3% (w/v) NaCl, pH 9.0이었으며, 종배양액 0.5% 접종시 40℃에서 24시간 배양했을 때 효소 생산량이 가장 높았다. Bacillus sp. DK1122가 생산하는 alkaline protease를 70% 포화 ammonium sulfate로 침전시키고, CM-Sepharose column chromatography에 의해 23.9%의 수율에 2.8배의 정제도를 지니는 효소를 얻을 수 있었다. SDS-PAGE를 통해 정제된 protease는 27 kDa의 크기의 단일 subunit으로 확인되었고, 정제된 효소의 최적 pH는 9.0, 최적온도는 60℃였으며, 50℃에서 1시간까지 열에 안정하였고, 60℃에서 10 mM CaCl₂ 첨가 후 3시간까지 90%의 활성을 유지하여 Ca²⁺에 의해 열안정성이 증가하였다. 본 연구를 통해 정제된 호알칼리성 protease는 식품, 세제 및 관련산업에서의 응용성이 매우 높을 것으로 기대된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권10호
• /
• pp.1416-1424
• /
• 2017

Objective: The objective of this study was to examine the effects of alfalfa flavonoids on the production performance, immunity, and ruminal fermentation of dairy cows. Methods: The experiments employed four primiparous Holstein cows fitted with ruminal cannulas, and used a $4{\times}4$ Latin square design. Cattle were fed total mixed ration supplemented with 0 (control group, Con), 20, 60, or 100 mg of alfalfa flavonoids extract (AFE) per kg of dairy cow body weight (BW). Results: The feed intake of the group receiving 60 mg/kg BW of AFE were significantly higher (p<0.05) than that of the group receiving 100 mg/kg BW. Milk yields and the fat, protein and lactose of milk were unaffected by AFE, while the total solids content of milk reduced (p = 0.05) linearly as AFE supplementation was increased. The somatic cell count of milk in group receiving 60 mg/kg BW of AFE was significantly lower (p<0.05) than that of the control group. Apparent total-tract digestibility of neutral detergent fiber and crude protein showed a tendency to increase (0.05<$p{\leq}0.10$) with ingestion of AFE. Methane dicarboxylic aldehyde concentration decreased (p = 0.03) linearly, whereas superoxide dismutase activity showed a tendency to increase (p = 0.10) quadratically, with increasing levels of AFE supplementation. The lymphocyte count and the proportion of lymphocytes decreased (p = 0.03) linearly, whereas the proportion of neutrophil granulocytes increased (p = 0.01) linearly with increasing levels of dietary AFE supplementation. The valeric acid/total volatile fatty acid (TVFA) ratio was increased (p = 0.01) linearly with increasing of the level of AFE supplementation, the other ruminal fermentation parameters were not affected by AFE supplementation. Relative levels of the rumen microbe Ruminococcus flavefaciens tended to decrease (p = 0.09) quadratically, whereas those of Butyrivibrio fibrisolvens showed a tendency to increase (p = 0.07) quadratically in response to AFE supplementation. Conclusion: The results of this study demonstrate that AFE supplementation can alter composition of milk, and may also have an increase tendency of nutrient digestion by regulating populations of microbes in the rumen, improve antioxidant properties by increasing antioxidant enzyme activities, and affect immunity by altering the proportions of lymphocyte and neutrophil granulocytes in dairy cows. The addition of 60 mg/kg BW of AFE to the diet of dairy cows was shown to be beneficial in this study.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권12호
• /
• pp.1764-1772
• /
• 2017

Objective: This study investigated the effects of different dietary energy sources on early postmortem muscle metabolism of finishing pigs. Methods: Seventy-two barrow ($Duroc{\times}Landrace{\times}Yorkshire$, DLY) pigs ($65.0{\pm}2.0kg$) were allotted to three iso-energetic and iso-nitrogenous diets: A (44.1% starch, 5.9% crude fat, and 12.6% neutral detergent fibre [NDF]), B (37.6% starch, 9.5% crude fat, and 15.4% NDF) or C (30.9% starch, 14.3% crude fat, and 17.8% NDF). After the duration of 28-day feeding experiment, 24 pigs (eight per treatment) were slaughtered and the M. longissimus lumborum (LL) samples at 45 min postmortem were collected. Results: Compared with diet A, diet C resulted in greater adenosine triphosphate and decreased phosphocreatine (PCr) concentrations, greater activity of creatine kinase and reduced percentage bound activities of hexokinase (HK), and pyruvate kinase (PK) in LL muscles (p<0.05). Moreover, diet C decreased the phosphor-AKT level and increased the hydroxy-hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) level, as well as decreased the bound protein expressions of HK II, PKM2, and lactate dehydrogenase A (p<0.05). Conclusion: Diet C with the lowest level of starch and the highest levels of fat and NDF could enhance the PCr utilization and attenuate glycolysis early postmortem in LL muscle of finishing pigs.

• 농약과학회지
• /
• 제5권2호
• /
• pp.1-12
• /
• 2001

다클론항체를 이용하여 환경 시료중 유기인계 살충제 acephate를 분석할 수 있는 경합적 간접효소면역측정법(competitive indirect ELISA)을 개발하기 위하여 분석물과 화학구조가 유사하며 hexanoic acid를 지닌 3종의 hapten을 합성하였다. 이들 hapten을 active ester 방법으로 담체단백질인 keyhole limpet hemocyanin과 접합시켜 면역원으로 사용하있고, bovine serum albumin과 접합시켜 homologous 혹은 heterologous ELISA를 위한 코팅항원으로 사용하였다. 생산된 항체들과 코팅항원을 최종선발하여 acephate 잔류분석을 위한 ELISA를 위하여 최적화하였다. 단백질의 농도가 $1{\mu}g/mL$인 코팅항원 (hapten-3-BSA)과 다클론항체 #8377를 16000배로 희석한 heterologous ELISA에서 최적화된 acephate의 $IC_{50}$ 값은 110 ng/mL이었고, 분석범위와 최소검출한계는 각각 $10{\sim}1000$과 4 ng/mL 이었다. Acephate의 주분해산물이고 살충제로 사용되는 methamidophos을 위시한 몇몇 유기인계 살충제의 항체에 대한 교차반응은 모두 0.02%이하였다. 이와 같은 결과는 본 ELISA가 농산물 및 환경시료중의 acephate 잔류분석을 위하여 편리한 방법이 될 수 있음을 시사한다.

• 한국식품위생안전성학회지
• /
• 제38권4호
• /
• pp.228-235
• /
• 2023

본 연구에서는 식품 제조시설 및 조리도구의 표면을 대상으로 식품알레르겐인 땅콩을 제거하기 위해 다양한 습식세척법을 평가하였다. 습식세척은 식품알레르겐 또는 잔류물질을 세척하는데 매우 효과적인 방법이며 식품 시설 또는 접촉 표면으로 사용되는 스테인리스, 나무, 플라스틱에 대해 온수로 5분간 불림 후 세척솔과 세척용제를 이용한 세척효과는 스테인리스> 유리> 플라스틱> 나무 순으로 확인되었다. 접촉 표면이 거친 나무는 끼임 등에 의해 세척효과가 낮게 나타났으며 표면의 세척효과를 높이기 위해 온수(50℃±2)로 5분간 불림 후 세척솔과 차아염소산 나트륨으로 세척하였을 때 나무표면으로부터 땅콩 알레르겐 제거에 효과가 큰 것으로 확인되었다. 식품 제조시설 및 조리도구의 표면에 따라 땅콩 알레르겐 세척효과는 달리 확인되어 본 연구에 사용된 재질과 세척용제 이외에 대해서도 추가의 연구는 필요하다. 식품안전현대화법에서 식품알레르기 표시 제도 및 식품알레르기 교차오염 예방 등에 관한 관리가 요구되고 있는 상황에서 식품 제조시설 및 조리도구 표면에 존재하는 식품알레르겐에 의한 교차오염의 예방과 관리는 필요할 것으로 생각된다. 본 연구 결과는 일반적인 세척 후에도 제조시설 및 도구의 표면에 땅콩 알레르겐의 잔류가능성을 제시하였고, 식품제조시설에서 취급하고 있는 땅콩 알레르겐의 세척 및 저감과 적절한 재질의 시설 및 도구 선정에 기초자료로 활용될 수 있을 것으로 판단된다.

• 한국환경농학회:학술대회논문집
• /
• 한국환경농학회 2009년도 정기총회 및 국제심포지엄
• /
• pp.273-282
• /
• 2009

The transmissible spongiform encephalopathies (TSEs) disease group are fatal neurodegenerative disorders affecting a wide range of hosts. The group includes kuru and Creutzfeldt-Jakob disease (CJD) in humans, scrapie in sheep and goats and Bovine spongiform encephalopathy (BSE) in cattle. The exact nature of the infectious agent involved in the transmission of these diseases remains controversial. However, a central event in their pathogenesis is the accumulation in infected tissues of an abnormal form of a host-encoded protein, the prion protein (PrP). Whereas the normal cellular protein is fully sensitive to protease ($PrP^{sen}$), the disease-associated prion protein ($PrP^d$) is only partly degraded ($PrP^{res}$), its amino-terminal end being removed. BSE was first reported in the mid-80s in the UK. Ten years later, a new form of human prion disease, variant CJD (vCJD) developed in the wake of the BSE epidemic, and there is now strong scientific evidence that vCJD was initiated by the exposure of humans to BSE-infected tissues, thus indicating a zoonotic disease. However, the ban on the feeding of animal-derived proteins to ruminants, and the apparent lack of vertical transmission of BSE, have led to a decline in the incidence of the disease within cattle herd and therefore, an assumed decreased risk for human contacting vCJD. The origin of the original case(s) of BSE still remains an enigma even though three hypotheses have been raised. Hypotheses are i) sheep- or goat-derived scrapie-infected tissues included in meat and bone meal fed to cattle, ii) a previously undetected sporadic or genetic bovine TSE contaminating cattle feed or iii) originating from a human TSE through animal feed contaminated with human remains. A host cellular membrane protein ($PrP^C$), which is abundant in central nervous system tissue, appear to be conformationally altered in the diseased host into a prion protein ($PrP^{Sc}$). This $PrP^{Sc}$ is detergent insoluble and partially protease-resistant ($PrP^{res}$). The term $PrP^{res}$ is normally used to describe the protein detected after protease treatment, in techniques such as Western immunoblotting, and enzyme-linked immunosorbant assay using fresh/frozen tissue. Immunohistochemistry may performed with formalin-fixed tissues. Also, clinical signs of the BSE are one of the major diagnostic indicators. Recently, atypical forms (known as H- and L-type) of BSE have appeared in several European countries, Japan, Canada and the United States. An unusual case was also reported in a miniature zebu. The atypical BSE fall into two groups based on the relative molecular mass (Mm) of the unglycosylated $PrP^{res}$ band relative to that of classical BSE, one of the higher Mm (H-type) and the other lower (L-type). Both types have been detected worldwide as rare cases in older animals, at a low prevalence consistent with the possibility of sporadic forms of prion diseases in cattle. This raises the unwelcome possibility that vCJD could increase in the human population. Now, active surveillance program against BSE is going on in Korea. In regional veterinary service lab, ELISA is applied to screen the BSE in slaughter and confirmatory tests by Western immunoblotting and immunohistochemisty are carried out if there are positive or suspect in the screening test. Also, the ruminant feed ban is rigorously enforced. Removal of specified risk materials such as brain and spinal cord from cattle is mandatory process at slaughter to prevent the infected material from entering the human food chain.