• Asian-Australasian Journal of Animal Sciences
• /
• 제25권7호
• /
• pp.1021-1028
• /
• 2012

Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose- and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan ($10{\mu}g/ml$, $20{\mu}g/ml$, $30{\mu}g/ml$, $40{\mu}g/ml$ and $50{\mu}g/ml$) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond $30{\mu}g/ml$ dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-${\alpha}$ and IFN-${\gamma}$ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.

• 한국식품저장유통학회지
• /
• 제23권6호
• /
• pp.765-771
• /
• 2016

전통적인 저온숙성 당절임 방법을 이용한 오미자청의 제조과정에서 최적의 배양시간을 찾기 위하여 파일럿 규모(1톤)의 연구를 시행하였다. 생오미자와 동일한 량으로 투입된 설탕은 배양시간에 따라 과당과 포도당으로 전환되었으며 오미자청에 존재하는 유리당의 조성은 68일 이상의 배양기간에서는 일정하게 유지되었다. 배양기간에 따른 오미자청의 pH는 일정한 수준을 유지한 반면, 적정 산도와 유기산의 함량은 배양시간 37일까지 지속적으로 증가한 이후 68일 이후에는 일정한 조성을 유지하였다. 생오미자에 존재하는 주요 유기산은 숙신산으로 확인되었으며 오미자청에 존재하는 유기산의 조성 역시 생오미자와 유사하였다. 오미자에 존재하는 주요 생리활성물질인 총 플라보노이드와 폴리페놀화합물은 60일 이상 배양한 오미자청에서 높은 함량을 보이고 138일까지 일정한 수준을 유지하였다. 특히, 오미자청에 함유된 총 플라보노이드와 폴리페놀화합물의 함량은 생오미자 과육에 비하여 각각 9배와 5배 정도 높은 수준으로 증가하였다. 생오미자를 저온숙성 당절임 방법을 적용하여 오미자청으로 제조 가공하는 과정에서 유리당, 유기산 및 생리활성물질 등에 대한 일정한 품질을 확보를 위해서는 최소 60일 이상의 배양기간이 필요한 것으로 확인되었다. 이러한 연구결과는 전통적인 저온숙성 당절임 방법으로 제조 가공되는 오미자청의 고품질화와 기능성 음료 개발을 위한 제조 가공공정의 표준화에 유용한 정보가 될 것으로 판단된다.

• 한국유기농업학회지
• /
• 제20권3호
• /
• pp.327-343
• /
• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• Journal of Animal Science and Technology
• /
• 제46권3호
• /
• pp.355-372
• /
• 2004

식물성유 첨가시 배양시간에 따른 지방산염의 형성 정도와 발효성상 및 NDF 소실율에 미치는 영향을 규명하고자 in vitro 시험을 실시 하였다. 기질은 알팔파 건초로 하고 1) oil 무첨가구, 2) 대두유 10%첨가구 및 3) 옥수수유 10%첨가구로 하였으며, oil 첨가 수준은 기질중량 건물 기준으로 첨가하였다. 발효성상 및 NDF 소실율을 위한 시험에서는 120ml serum bottle을 사용하여 3반복 실시하여 pH, $NH_3-N$, VFA농도 및 건물과 NDF 소실율을 분석하엿다. 지방산 분획을 위한 시험에서는 60ml serum bottle을 사용하요 3반복 실시하여 NEFA, EFA 및 FAS의 형성 정도를 분석하였다. pH는 배양시간이 증가함에 따라 유읭적으로 감소하였고, $NH_3-N$ 농도는 유의하게 증가하였다(P<0.0001). 건물 및 NDF 소실율은 처리구, 배양시간 및 oil간 유의하게 차이를 보여주었다(P<0.05). 총 휘발성 지방산 농도는 배양시간이 증가함에 따라 유의하게 증가하였다(P<0.0001). Acetate는 시험구 모두 배양시간이 증가함에 따라 유의적으로 감소하였고(P<0.0001), 대조구에 비해 oil 첨가구가 유의적으로 낮아졌다(P<0.05). Propionate는 시험구 모두 배양시간이 증가함에 따라 유의적으로 증가하였지만(P<0.0001), 처리구 및 oil간 효과는 없었다. Oil 첨가구에서 A/P비율이 유의하게 낮아졌다(P<0.05). EFA는 배양시간이 증가함에 따라 유의하게 감소하였고(P<0.0001), NEFA는 EFA의 가수분해로 인하여 증가하였다. 배양시간 48시간에서는 NEFA가 양이온과 결합하여 FAS의 형성 비율이 유의적으로 증가하였다(P<0.0001). 특히 $C_{18:0)$의 FAS 형성은 배양시간 48시간에서 대두유 및 옥수수유 첨가구에서 각각 12.53 및 15.17mg/g DM으로 0시간에 비해 각각 27.5 및 32.5배 증가하였다(P<0.0001). 배양시간이 증가할수록 칼슘을 포함한 양이온 함량이 높은 알팔파가 분해되면서 양이온이 용해되어 유리된 지방산과 결합하여 지방산염을 형성하게 된다. 이때 불포화지방산보다는 포화지방산이 양이온과 반응하여 지방산염을 형성하기 쉽다. 또한 $C_{16:0}$ 이상의 장쇄지방산이 주로 지방산염을 형성하였다. 이러한 특성 때문에 배양액내에 유리된 지방산이 염을 형성하기 쉽고 oil 첨가시 발생하는 발효성상 및 NDF소실율에 미치는 부의 영향을 최소화 한 것으로 사료된다.

• Animal Bioscience
• /
• 제37권5호
• /
• pp.908-917
• /
• 2024

Objective: Although pork loins is not a tough meat, they need to develop meat products with a soft texture for the elderly. This study focused on the physicochemical properties and tenderness characteristics of pork loin injected with green kiwifruit juice (GRJ) and gold kiwifruit juice (GOJ) during various incubation times. In addition, the antioxidant activities of hydrolysate derived from the hydrolysis of pork loin by kiwifruit juice protease were evaluated. Methods: The pork loin was injected with 10% and 20% GRJ and GOJ, under various incubation times (0, 4, 8, and 24 h). Then, the physicochemical properties and tenderness of pork loins were measured. 2,2- diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power were conducted to determine hydrolysate's antioxidant activities derived from pork loin's hydrolysis by kiwifruit juice protease. Results: GRJ had greater tenderizing ability than GOJ, even at the 10% addition. When kiwifruit juice was injected into pork loin, the tenderness increased with increasing incubation time. This was confirmed by the decrease in intensity of the myosin heavy chain (MHC) band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In particular, the MHC band decreased at 8 h for both 10% GRJ and 20% GOJ and at 4 h for 20% GRJ alone. The highest myofibril fragmentation index and peptide solubility were observed in pork loin treated with 20% GRJ compared to the other treatments during incubation. The 10% GRJ and 20% GOJ treatments showed similar levels of antioxidant activity of the protein hydrolysates in pork loin, and 20% GRJ showed the highest activity among the treatments. Conclusion: Kiwifruit juice had protease activity, and GRJ was more useful for tenderizing meat products than GOJ. Thus, GRJ at 10% could be a potential agent to tenderize and enrich the natural antioxidant activity through the proteolysis of pork loin.

• Archives of Pharmacal Research
• /
• 제18권3호
• /
• pp.153-158
• /
• 1995

The levels of extracellualr glutamate, intracellular $Ca^{2+}\;([Ca2+]_i)$ and cGMP were determined for 1 h with the excitatory amino acids, N-methyl-D-aspartate (NMDA) or kainate in cultured cerebellar granule cells. Both NMDA and kainate produced a time-dependent release of glutamate, and kainate was more potent than NMDA in glutamate elevation. The elevation of extracellular glutamate was not purely governed by intracellular $Ca^{2+}$ concentration. However, in opposite to the time-dependent elevation of glutamate, the elevation of cGMP by NMDA and kainate were at maximum level in short-time (1 min) incubation then remarkably decreased with longer incubation times. Post-applications (30 min after agonist) of EAA antagonist did not block EAAs-induced glutamate elevation. However, NMDA antagonist, phencyclidine (PCP), blocked NMDA-induced cGMP elevation at pre- or post-application, but kainate antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX), paradoxically augmented kainate-induced cGMP elevation for 1 h incubation. These results show that NMDA or kainate induces time-dependent elevations of extracellular glutamate, while the elevations of cGMP by these EAAs are remarkably decreased with longer incubation times. However, NMDA- arid kainate-indcued glutamate release was blocked by pre-application of each receptor antagonist but not by post-application while EAA-induced $[Ca^{2+}]_i$ was blocked by post-application of antagonist. These observations suggest that EAA-induced elevation of $[Ca^{2+}]_i$ is not parallel with elevation of glutamate release or cGMP.

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2004년도 춘계학술발표대회
• /
• pp.256-256
• /
• 2004

The study was carried out to investigate the effects of preservation of ovaries and oocytes with and without cumulus cells and incubation time on zona penetration by canine spermatozoa. The objective of this study was to produce in vitro fertilized oocytes and solute canine sterile. (omitted)

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권5호
• /
• pp.752-757
• /
• 1999

Studies were made of digestion of timothy (Pheleum pretense) hay, tall fescue (Festuca elatior) hay, and rice (Oryza sativa) straw in pure cultures of rumen anaerobic fungus, Neocallimastix frontails MCH3. The fungus was inoculated on ground forages (1%, w/v) in an anaerobic medium and incubated at $39^{\circ}C$. Incubation was continued for 24, 48, 72 and 96 h. The losses of dry matter, xylose and glucose of forage during incubation were determined at the end of these incubation periods. Xylose and glucose were considered to be released from xylan and cellulose, respectively. The digested xylan to digested cellulose (X/C) ratios of the substrate were calculated. Xylanase and carboxymethyl cellulose (CMCase) of culture supernatant and residual substrate was measured at the same time. The X/C ratios in the cultures on timothy hay and rice straw were greater than 0.5 in the first 24-h incubation period. The values were smaller than 0.3 in tall fesque. The ratio of xylanase activity to that of CMCase in the first 24-h incubation period correlated well with the traits in X/C ratio. However xylanase activity was still superior to CMCase in the following incubation period (48 to 96 h), although the glucose (designated as cellulose) was more intensively digested than xylose (designated as xylan). The production of these polysaccharidases appeared to correlate with substrate cell-wall sugar composition, xylose to glucose ratios, at the beginning of fast growing period.

• Journal of Dairy Science and Biotechnology
• /
• 제41권2호
• /
• pp.45-56
• /
• 2023

This review serves two purposes: first, to promote the use of improved optimization techniques in response surface methodology (RSM); and second, to enhance the optimum conditions for the fermentation of probiotics. According to research in dairy science, Lactiplantibacillus plantarum K79 is a candidate probiotic that has beneficial health effects, such as lowering blood pressure. The optimum conditions for L. plantarumK79 to produce peptides with angiotensin-converting enzyme (ACE) inhibitory activity were proposed, through modeling each of ACE inhibitory activity and pH as a function of the four factors that are skim milk concentration (%), incubation temperature (℃), incubation time (hours), and starter added amount (%). To estimate optimum conditions, the researchers employed a desirability-based multi-response optimization approach, utilizing third-order models with a nonsignificant lack of fit. The estimated optimum fermentation conditions for L. plantarum K79 were as follows: a skim milk concentration of 10.76%, an incubation temperature of 36.9℃, an incubation time of 23.76 hours, and a starter added amount of 0.098%. Under these conditions, the predicted ACE inhibitory activity was 91.047%, and the predicted pH was 4.6. These predicted values achieved the objectives of the multi-response optimization in this study.

• 한국수정란이식학회지
• /
• 제26권4호
• /
• pp.251-256
• /
• 2011

Presently, the effect of 0.5 mM dibutyryl cAMP (dbcAMP)-supplemented maturation medium during different incubation time on meiotic arrest (germinal vesicle) and resumption (metaphase II) of porcine oocytes and embryonic development of porcine oocytes following in vitro fertilization (IVF) or parthenogenetic activation (PA) was determined. Porcine cumulus oocyte complexes (COCs) were cultured in 0.5 mM dbcAMP for 17, 22, 27, or 42 h, and an additional 22 h without 0.5 mM dbcAMP. The nuclear status was examined at each time point. Oocytes cultured from 39~49 h displayed more than 80% meiotic resumption. More than 85 % of meiotic arrest was presented at 17~22 h. Oocytes were cultured for 22 h with 0.5 mM dbcAMP and additional 22 h without dbcAMP to assess developmental potential following IVF or PA. There were no significant differences in blastocyst rates among the dbcAMPIVF, IVF, dbcAMP-PA, and PA groups, although cleavage rate of IVF group was significantly higher than those of dbcAMP-PA, and PA groups. In conclusion, 0.5 mM dbcAMP influenced meiotic maturation of porcine oocytes depending on incubation time of oocyte, although embryonic development was not improved in both IVF and PA.