Antioxidants, as reactive oxygen species scavengers, are one of the beneficial additives in serum-free defined culture medium. In this study, three separate experiments were performed to determine the effects of 3-hyroxyflavone added to the culture medium on the developmental competence of follicular bovine oocytes during in vitro maturation (IVM) and/or in vitro culture (IVC). The rate of blastocyst developed from oocytes cultured in IVM medium with 3 hyroxyflavone was significantly higher than that from control oocytes (39.0% vs. 26.3%, p<0.001), respectively. However, oocytes cultured in the medium with addition of 3-hyroxyflavone only at IVC period did not show significance in the blastocyst development when compared with control. When 3-hyroxyflavone was added to both IVM and IVC media, the rate of blastocyst formation was even significantly lower (21.1%) than control (26.5%; p<0.05). The present findings suggested that antioxidative activity of 3-hydroxyflavone added to only IVM medium beneficially affected the developmental competence of follicular bovine

본 연구에서는 메탄올과 열수를 이용해 갈색먹물버섯의 자실체로부터 추출한 물질의 항산화, 항당뇨, 항콜린에스 테라아제, 항티로시나제와 항염증 효과를 탐색하 였다. 고 속액체크로마토그래피를 이용해 추출물의 페놀성 화합물 을 분석한 결과 procatechuic acid, chlorogenic acid, (-)- epicatechin, naringin 등 총 4종류의 페놀성 화합물이 확 인되었다. 항산화 효과 실험에서 DPPH radical 소거능은 양성대조군으로 사용한 BHT에 비해 낮았지만 효과가 비 교적 우수하였고, 철 이온을 제거하는 항산화 효과는 메 탄올과 열수 추출물이 양성대조군인 BHT에 비해 30% 높게 나타났으나 환원력은 양성대조군에 비해 43% 정도 낮은 것으로 나타났다. 항당뇨 실험에서 α-amylase와 α- glucosidase에 대한 메탄올과 열수추출물의 저해효과는 2.0 mg/ml의 농도에서 각각 62.26%와 67.59%를 보여 양 성대조군인 acarbose의 81.81%에 비해 낮았다. 아세틸콜 린에스테라아제에 대한 메탄올과 열수추출물의 저해효과 는 1.0 mg/ml의 농도에서 각각 94.64%와 74.19%를 보여 양성대조군인 galanthamine의 97.80%에 비해 낮았다. 티 로시나아제에 대한 메탄올과 열수추출물의 저해효과는 2.0 mg/ml의 농도에서 각각 91.33%와 91.99%를 나타내 양성대조군인 kojic acid의 99.61%와 매우 유사한 효과를 얻었다. 염증저해 효과 실험에서는 RAW 264.7 대식세포 가 배양되고 있는 배지에 갈색먹물버섯 자실체의 메탄올 과 열수추출물을 각각 전 처리 한 후 염증매개 물질인 LPS를 투여하여 메탄올과 열수 추출물의 NO 생성 저해 효과를 조사한 결과 추출물의 농도가 증가함에 따라 생성 된 NO의 양이 감소하는 경향을 나타내었다. 따라서 갈색 먹물버섯의 자실체에는 항산화, 항당뇨, 항아세틸콜린에 스터라제 항티로시나아제 및 항염증 효과를 나타내는 물 질이 함유되어 있어서 천연 건강식품으로 이용이 가능할 것으로 사료된다.

실험에서는 꽃송이버섯 (Sparassis crispa, formerly S. crispa) 에탄올 추출물의 소수성 분획을 분리하고 각 분획의 DPPH 항산화 활성과 위암 (AGS), 폐암 (A529), 간암 (HepG2) 세포주를 이용한 항암 활성을 MTT assay 를 통해 확인 하였다. SOCC를 사용하여 총 18개의 fraction으로 분획하였고 TLC와 세포주를 이용한 항암 활 성 확인을 통하여 5개의 fraction으로 압축하였다. 항암 활성이 높은 5개의 fraction은 HPLC-MS를 통해 각 분획 물을 분석한 결과 항산화 활성을 보이지 않았으며 약 181.0의 분자량을 가진 물질이 지표물질로 확인되었으므 로 이 물질의 화학식 동정을 위하여 추가실험이 필요하다. 세포주를 이용한 항암실험 결과 꽃송이버섯 추출물은 위 암 (AGS), 폐암 (A529), 간암 (HepG2) 세포주 모두에서 양성대조군인 paclitaxel보다 낮은 세포 생존율을 보여 주 었으며(IC50 value), 이것은 추후 꽃송이버섯 추출물에 포 함된 항암 물질 분리 연구를 위한 기초연구 결과로서 무 척 의미가 크다고 할 수 있다.

전 세계적으로 최근 아토피 피부염의 유병율이 증가하면서 이 문제에 대한 관심이 증가하고 있지만 근본적으로 개선시킬 수 있는 효과적인 치료법이 개발되지 못한 상태로 단순히 증상을 경감시키는 대증요법인 스테로이드 같은 약물치료만이 현재 표준 치료로 시행되고 있는 실정이다. 이러한 약물치료는 장기적인 측면에서 내성 증가와 증상의 악화 등의 문제점으로 인해 최근에는 천연성분을 이용한 항아토피 소재에 대한 연구들이 활발히 진행되고 있다. 그 중 식용버섯인 Hypsizigus marmoreus(HM, 느티만가닥 버섯)는 많은 생리활성 물질을 포함하고 있으며 특히, 항염효과가 우수한 것으로 밝혀져 있는데 본 연구에서는 HM이 아토피 피부염에 미치는 효과를 측정하였다. 동결건조한 버섯 시료를 85% 에탄올로 추출하여 추출물을 제조하였으며, 이를 동결건조하여 시료로 사용하였다. 마우스 유래의 대식세포인 RAW264.7에 염증 유발 물질인 LPS(lipopolysaccharide)와 HM 추출물을 처리하여 염증 관련 싸이토카인 TNF-α, IL-6, IL-2, IL-4 및 IFN-γ과 nitric oxide(NO)생성량을 확인하였다. RAW264.7 세포주에 HM 추출물을 농도별로 처리한 결과, 고농도로 처리된에서 높은 NO 생성 저해능을 보여주었다.

In this study we conducted to rear worker honey bee (Apis cerana) from larvae to adult stage in the laboratory by using plastic well plates. Our study results were showed that honey bee larvae Apis cerana could be reared in the laboratory. The adult worker bee started to emerge on day 17 from grafting. The emergence of worker bee peak on day 18 and declined thereafter. The average survival rate from larvae to pre-pupae stage was 74.6%. The average survival rates from pre-pupae to adult stage and from larvae to adult stage were 40.7 % and 30.4 % respectively.

Cockroaches are insects of the order Blattodea, sometimes also called Blattaria, of which about 30 species out of 4,600 total are associated with human habitats. About four species are well known as pests. Among the best-known pest species are the American cockroach, Periplaneta americana, which is about 30 mm long; the German cockroach, Blattella germanica, about 15 mm long. We researched growing condition of in-vitro(temperature and humidity) for P. americana and B. germanica. The cockroach is divided in three sections; the body is flattened and broadly oval, with a shield-like pronotum covering its head. A pronotum is a plate-like structure that covers all or part of the dorsal surface of the thorax of certain insects. They also have chewing mouth parts, long, segmented antennae, and leathery fore wings with delicate hind wings. The third section of the cockroach is the abdomen. We measured size of head, abdomen, and body length during differential stages. Also, we checked number of egg, size of egg, hatching rates, period of former laying eggs, and laying periods etc.

The aim of the present study was to compare two different serum-free media, modified synthetic oviduct fluid (mSOF) and modified potassium simplex optimization medium (mKSOM) containing 20% RD (RPMI1640 + DMEM, 1:1 v/v) (RD-mKSOM), for in vitro culture (IVC) of bovine embryos. After in vitro maturation and fertilization, the presumptive zygotes were cultured in two different serum-free conditions for 7 days and 9 days to evaluate blastocyst formation and hatching, respectively. Serum supplemented conventional CR2 medium was used as control. After 7 day of culture, there was no significant difference in cleavage and blastocyst formation rates among three groups (mSOF, 59.3 and 30.1%; RD-mKSOM, 65.0 and 41.5%; control, 51.6 and 38.0%, respectively). Hatching rate was significantly higher in control (69.0%) than other experimental groups (mSOF, 22.0%; RD-mKSOM, 39.5%) (P＜0.0001 and P＜0.001, respectively). Although both serum-free conditions showed lower hatching rates than serum-added control, in serum-free groups, RD-mKSOM showed significantly higher hatching rate than mSOF (P＜0.001). In addition, one-step using RD-mKSOM may facilitate IVC procedure than two-step culture system. In conclusion, the results indicate that one-step RD-mKSOM is more suitable defined culture system for IVC of bovine embryos than two-step mSOF.

This study was conducted to examine the effects of defaunation (removal of live protozoa) on fermentation characteristics, degradation of ryegrass hay and CH4 (methane) production by rumen microbes when incubated with plant oils (SO, sunflower oil and LO, linseed oil) in vitro. Sodium lauryl sulfate (0.000375 g/ml) as a defaunation reagent was added into the culture solution and incubated anaerobically up to 24 h at 39℃. pH from defaunation was increased for all treatments from 6 h incubation times (p<0.01-0.001) compared with those from fauantion. Concentration of ammonia-N from defaunation is higher than that from faunation at 3 h (p<0.001), 12 h (p<0.05) and 24 h (p<0.001) incubation times. Defaunation decreased (p<0.01-0.001) total volatile fatty acid concentration at all incubation times. Molar proportions of C2 (acetate, p<0.05-0.001) and butyrate (p<0.01-0.001) were also decreased by defaunation at all incubation times. Molar proportion of C3 (propionate), however, was increased by defaunation at all incubation times (p<0.001). Thus the rate of C2 to C3 was decreased by defaunation at all incubation times (p<0.001). Defaunation decreased ED (effective degradability) of dry matter (p<0.001) and ED of neutral detergent fiber (p<0.001) of ryegrass hay. Defaunation decreased total gas, CH4 production, CH4 % in total gas and CH4/CO2 at all incubation times (p<0.001). Oil supplementation decreased total gas (p<0.05-0.001), CH4 production (p<0.001) and CH4 % in total gas (p<0.001) compared with control at all incubation times. The result of this study showed that defaunation combined with oil supplementation may cause an alteration of microbial communities and further medicate the fermentation pattern, resulting in both reduction of degradation of ryegrass hay and CH4 production. No difference, however, was observed in all the examinations between SO and LO.

During bone remodeling, there is requirement of differentiation of osteoblastic cells. Previously, we identified proteins differentially expressed in soft tissue during bone healing. Of these proteins, we focused the effect of LTF on differentiation of osteoblast. In order to analyze the osteogenic ability of LTF, we treated conditioned media collected from human LTF-stably transfected HEK293T cells into osteoblastic MC3T3-E1. The results showed that the activity and expression of alkaline phosphatase were increased in MC3T3-E1 cells treated with conditioned media containing LTF in dose- and time-dependent manner. At the same time, we observed the significant increase of the expression of osteoblastic genes, such as ALP, BSP, COL1A1, and OCN, and along with matrix mineralization genes, such as DMP1 and DMP2, in LTF conditioned media-treated groups. Moreover, the result of treating recombinant human LTF directly into osteoblastic MC3T3-E1 showed the same pattern of treating conditioned media containing LTF. Our study demonstrated that LTF constitutively enhances osteoblastic differentiation via induction of osteoblastic genes and activation of matrix mineralization in MC3T3-E1 cells.

본 연구는 오배자 에탄올 추출물 (GRE), 염소산나트륨 (SC) 그리고 오배자 에탄올 추출물과 염소산나트륨 합제(GS)의 B. abortus에 대한 항균효과를 확인하기 위해 수행 되었다. GRE, SC 그리고 GS를 B. abortus에 처리하여 배양한 후, B. abortus의 생존수를 확인하였으며, 마우스 탐식세포 내 감염된 B. abortus의 증식 억제효과를 경시별 (2, 24, 48시간)로 조사하였다. GRE, SC 그리고 GS는 각각 400 μg/mL 이하, 15 mM 그리고 0.6GS (GS 1, GRE 1,000 μg/mL + SC 30 mM) 이하의 농도에서 세포독성을 나타나지 않았다. 모든 처리구에서 B. abortus의 생존율은 용량- 의존적으로 현저하게 감소하는 결과를 나타내었다. 또한, GRE (400 μg/mL), SC (15 mM) 그리고 0.5GS (GRE 500 μg/mL + SC 15 mM)를 처리한 세포에서 배양 48시간 후에, B. abortus의 증식이 통계적으로 유의성 있게 감소하였으며 (GRE, p < 0.01; SC and 0.5GS, p < 0.001), 특히, GS를 처리한 경우, B. abortus의 세포내 증식이 GRE와 SC의 상승작용에 의한 강력한 항균효과를 나타내었다. 결론적으로, GS는 B. abortus에 대한 항균물질로서 유용할 뿐만 아니라, 식육과 우유 위생 분야에 적용할 수 있을 것으로 생각된다.

The objective of this study was to investigate the changes of oxidative stress and antioxidant enzyme during in vitro development with washing culture oil in porcine embryos. During the culture, the four types of culture oil such as paraffin oil with or without washing and mineral oil with or without washing were examined. The oil was washed with PZM-3 during 7 days and collected oil only. The embryos were stained with CellTrackerTM Red, DC-FDA and Hoechst 33342 to confirm the effects of the oil. As a results, Cleavage rates and total cell number were no difference among the four oil groups. However, ≥16 cell embryos were significantly different in fore type oil treat-ment and blastocyst rate was significantly higher washing paraffin treatment than in other group(p<0.05). Also, the expression of free radical were lower in washing paraffin oil than in other groups (p<0.05). On the other hand, the expression of glutathione were not significant different among paraffin oil with or without washing and mineral oil with or without washing, however washing paraffin oil and washing mineral groups were higher than other treat-ment groups. In conclusion, the washing oil was expected with positive effects on in vitro development in porcine embryos.

In this experiment, a highly porous scaffold of biphasic calcium phosphate (BCP) was prepared using the spongereplica method. The BCP scaffold was coated with 58S bioactive glass (BG) and sintered for a second time. The resulting scaffold was coated with gelatin (Gel) and cross-linked with [3-(3-dimethyl aminopropyl) carbodiimide] and N-Hydroxysuccinamide (EDC-NHS). The initial average pore size of the scaffold ranged from 300 to 700μm, with more than 85 % porosity. The coating of BG and Gel had a significant effect on the scaffold-pore size, decreasing scaffold porosity while increasing mechanical strength. The material and surface properties were evaluated by means of several experiments involving scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffraction (XRD). Cytotoxicity was evaluated using MTT assay and confocal imaging of MC3T3-E1 pre-osteoblast cells cultured in vitro. Three types of scaffold (BCP, BCP-BG and BCP-BG-Gel) were implanted in a rat skull for in vivo evaluation. After 8 weeks of implantation, bone regeneration occurred in all three types of sample. Interestingly, regeneration was found to be greater (geometrically and physiologically) for neat BCP scaffolds than for two other kinds of composite scaffolds. However, the other two types of scaffolds were still better than the control (i.e., defect without treatment).

본 시험은 효소제 및 효모추출물 급여가 사양성적, in vitro 반추위 발효 및 혈액성상에 미치는 영향을 구명하기 위해 23개월령의 한우 거세우 48두를 공시하여 사료용 첨가제로 농후사료의 0.1% 수준으로 급여하여 출하시까지 대조구(무첨가), 처리구 1(효소제), 처리구 2(효모추출물), 처리구 3(효소제+효모추출물)으로 구분하여 급여하였다. 사양성적에서는 급여 4개월과 8개월의 일당증체량과 사료요구율이 처리구 1에서 가장 향상되었고 (p<0.05), 대조구에서 가장 낮았다(p<0.05). 도축성적에서는 육량지수를 포함하여 처리구 1>처리구 3>처리구 2>대조구 순으로 나타났다. in vitro 배양액에 따른 처리구별 pH는 대조구>처리구 2>처리구 3>처리구 1 순으로 유의적(p<0.05)인 차이를 나타냈고, VFA와 사료소화율은 처리구 1과 3에서 높았고 (p<0.05), 대조구에서는 낮았다(p<0.05). 혈액성상에서는 면역 첨가제 급여군인 처리구 2와 3에서 급여기간에 따른 백혈구 수치가 유의적(p<0.05)으로 감소하였는데, 이는 면역글로블린 G의 유의적(p<0.05)인증가에 따라서 상대적으로 나타난 것으로 판단된다. 결과적으로, 한우 비육후기 거세우의 사료첨가제 급여 시험에서 처리구 1은 급여 4개월부터 대조구 대비 사양성적 및 도체성적을 향상시켰고, 처리구 2는 급여 2개월부터 면역에 관련된 면역글로블린 G의 혈액성상을 개선시키는 것으로 분석되었다.

K+ channels are involved in the regulation of a variety of physiological functions, including proliferation, apoptosis and differentiation, in mammalian cells. Our previous study demonstrated that the blockage of K+ channels inhibits mouse early embryonic development. This study was designed to identify the effect of K+ channels during bovine embryonic development. K+ channel blockers (tetraethylammonium (TEA), BaCl2, quinine, ruthenium red and fluoxetine) were added to the culture medium during in vitro fertilization (IVF) for 6 h to first identify the short-term effect of these chemicals. Among K+ channel blockers, fluoxetine, which is used as a selective serotonin reuptake inhibitor, significantly increased the blastocyst formation rate by approximately 6% when compared to control. During the in vitro maturation (IVM) of immature oocytes and the in vitro culture (IVC) of embryos, the oocytes and embryos were exposed to fluoxetine for either a short-term (6 h) or a long-term (24 h) to compare the embryonic development in response to exposure time. The 6 h exposure to fluoxetine during IVM did not affect the blastocyst formation rate, but the rate of blastocyst formation was reduced after the 24 h exposure. On the other hand, embryonic development increased approximately 10% in both groups of embryos exposed to fluoxetine for 6 and 24 h during IVC. Taken together, fluoxetine treatment during IVF and IVC, but not IVM, enhances bovine embryonic development. These results suggest that fluoxetine-modulated signals in oocytes and embryos could be an important factor towards enhancing bovine embryonic development.

본 연구는 농․식품부산물 중 비지박, 사자발약쑥 그리고 커피박의 유효미생물(L. acidophilus ATCC 496, L. fermentum ATCC 1493, L. plantarum KCTC 1048, L. casei IFO 3533)발효 사료원이 젖소 급여용 TMR(대조구) 중 대두박을 주로 대체한 각 비지박(SC), 발효 비지박(FSC), 발효 비지박+발효 사자발약쑥 부산물(1:1, DM basis, FSCS) 3, 5, 10% 그리고 발효 비지박+발효 커피박(1:1, DM basis, FSCC) 3, 5, 10%의 반추위 내 미생물 발효특성을 알아보기 위해 9처리구를 이용하여 3반복으로 in vitro 시험이 실시되었다. 배양 6~8시간대에 FSCS와 FSCC 처리구는 SC와 FSC 처리구에 비해 높은 pH 수준을 유지하였고, 대조구에 비해 낮은 pH 수준을 유지하였다(p<0.05). 또한 배양 24시간대 FSC와 FSCC 3% 처리구는 가장 높은 pH를 유지하였다(p<0.05). Gas 생성량은 배양 4~10시간대까지 FSCS와 FSCC 처리구가 대조구 보다 유의적으로 높았으나 SC와 FSC 처리구 보다 낮은 결과를 나타내었다(p<0.05). 건물소화율은 배양 12시간대까지 FSC 처리구에서 유의적으로 높았고(p<0.05), 24시간대에는 FSCS 10% 처리구가 가장 높은 결과를 나타내었다(p<0.05). NH3-N 함량은 배양 4시간대까지 대조구에서 가장 낮았고(p<0.05), 24시간에서는 FSC 처리구가 가장 높은 결과를 나타내었다(p<0.05). 미생물단백질 합성량은 배양 시작 시 FSC, FSCS 그리고 FSCC처리구가 4종의 Lactobacillus spp.에 의한 발효 때문에 대조구와 SC 처리구에 비해 높았으며(p<0.05), 배양 10시간대 FSC 처리구 보다 SC, FSCS 그리고 FSCC 처리구에서 유의하게 높은 결과를 나타내었다(p<0.05). Total VFA 농도는 배양 6~12시간대에는 FSC 처리구에 유의하게 높았고(p<0.05), 배양 24시간대에는 FSCS 처리구에서 가장 높았다(p<0.05). Acetate 농도는 배양 0~12시간대에 FSC 처리구에서 가장 높았으며(p<0.05), 24시간대에는 FSCS 5, 10% 처리구에서 가장 높은 결과를 나타내었다(p<0.05). Propionate 농도는 FSC 처리구가 배양 0~10시간대까지 가장 높았으며(p<0.05), 전체 배양기간 동안 FSCS 5, 10% 처리구와 FSCC 처리구에 비해 높은 경향을 나타내었다. Acetate/Propionate 비율은 배양 12시간대에 SC 처리구에서 가장 높았으며, 배양 24시간대에는 FSCS 3% 처리구가 가장 낮은 결과를 나타내었다(p<0.05). CH4 가스 생성량은 FSC 처리구에서 배양 0~10시간대까지 가장 높았으며(p<0.05), 24시간대에는 가장 낮은 결과를 나타내었다(p<0.05). 배양 4~10시간대까지 FSCS와 FSCC 처리구는 SC와 FSC 처리구에 비해 유의하게 낮은 CH4 가스 생성량을 나타내었고(p<0.05), 사자발약쑥과 커피박 함량이 증가할수록 CH4 가스 생성량은 배양 종료 시까지 낮은 경향을 나타내었다.따라서, 본 in vitro 시험에서 젖소 급여용 TMR에 SC, FSC, FSCS 그리고 FSCC의 대체효과는 FSC 처리구에서 배양 초기부터 12시간대까지 반추위 내 미생물 발효 특성이 가장 높았다. FSCS와 FSCC 처리구는 배양 6~8시간대부터 반추위 내 미생물 작용이 높아지는 특징을 나타내었으나, 배양 시작 후 6~8시간대까지 gas 생성량 및 CH4 가스 발생량은 낮아지는 특징을 나타내었다. 이와 같은 결론은 FSC는 젖소용 TMR 사료원 대체 효과가 높으며, TMR사료원 중 FSCS와 FSCC 대체효과는 배양 초기 다소 낮은 반추위 내 미생물 발효 특성을 나타내었으나 CH4 저감효과를 나타내었다. 또한 배양 후기에는 SC와 FSC의 반추위 내 미생물 발효 특성과 비슷한 경향을 나타내어 TMR 사료원으로써 이용 가치가 높을 것으로 사료된다.

This study was carried out to investigate the effects of tissue inhibitor of matalloproteinase-1 (TIMP-1), Activin A and Heparin binding epidermal growth factor (HB-EGF) on in vitro production of bovine embryos. In experiment 1, presumptive zygotes were cultured in the medium supplemented with TIMP-1 (0.5 μg/ml), Activin A (100 ng/ml), or HB-EGF (100 ng/ml) at 39 ℃ in a humidified atmosphere of 5% (v/v) CO2, 5% (v/v) O2 and 90% (v/v) N2. In experiment 2, TIMP-1 + HB-EGF or Activin A + HB-EGF combinations were supplemented in the culture medium. The developmental rate to blastocysts, hatching rate and total cell numbers of the blastocysts were evaluated in both experiments. The embryos cultured in medium without growth factor supplementation was used as control group. In experiment 1, the embryos cultured in medium supplemented with TIMP-1 and Activin A showed significantly higher developmental rate to blastocysts than those cultured with HB-EGF and control (36.9%, 34.1%, 21.2% and 23.1%, respectively) (P<0.0001). However, the hatching rate of blastocyst was significantly higher in embryos with HB-EGF than those with TIMP-1, Actvin A and Control groups (84.4%, 58.8%, 51.4% and 49.3%, respectively) (P<0.001). Total cell number per blastocyst was also significantly higher in embryos with HB-EGF group (174.3±2.5) than those with TIMP-1, Activin A (149.7 and 150.0, respectively) (P<0.05) and Control (119.0) (P<0.001). In experiment 2, embryos cultured with combined treatment of Activin A and HB-EGF resulted in significantly higher rates of blastocysts formation (48.0%), hatching rate (89.7%) and total cell number in blastocyst (182.3±2.1) than those with TIMP-1 and HB-EGF combination group (32.0%, P<0.001; 76.6%, P<0.05; 165.7±4.2, P<0.001, respectively). Our data demonstrate that in vitro production of bovine embryos could be improved by combined supplementation of Activin A and HB-EGF in culture medium.

Matrix Metalloproteinases (both MMP2 and -9) play a pivotal role of the embryos hatching and implantation. Therefore, the objective of this study was carried out to investigate the influence of MMP2 and MMP9 on embryo development potential and subsequent effect at molecular level. There was no significant difference of cleavage rate among the groups. The development competence of blastocyst was significantly higher (P<0.05) in MMP9 treatment (39.81±16.61) than that to the combined treatment of MMP2 and –9 (23.68±0.27), but there was no significant difference among the control vs. MMP2 vs. MMP9 (35.05±2.74 vs. 32.71±6.18 vs. 39.81±16.61, respectively). On the other hand, the hatching rate of blastocysts was significantly lower (P<0.05) in combined group of MMP2 and –9 (12.55±0.09) (Table1). The expression level of MMP2 and MMP9 was significantly lower (P<0.05) in the entire treatment groups than that in the control group. But the expression of MMP9 was significantly higher (P<0.05) when compared in the entire treatment groups. The relative expression embryonic developmental gene, IFNt expression level significantly lower (P < 0.05) in the MMP9 embryos. The placenta establishment genes, PLAC8 and SSLP1, expression were significantly higher (P < 0.05) in the MMP2 embryos compared to other groups. Transcription regulation gene, HNRNPA2B1, was higher (P < 0.05) in the combined group of MMP2+MMP9 than that in the other groups. In conclusion, our results suggest that MMPs to culture medium improves the blastocyst development rate and further impact on target gene expression analysis.

The objective of this study was to know the in vitro effects of supplemental anthelmintic plant extracts on the inhibition of protozoa for reducing methane production in the rumen. A fistulated Holstein cow was used as a donor of rumen fluid. The plant extracts (Lonicera japonica, Zanthoxylum piperitum, Pyrethrum, Torreya nucifera, Ruta graveolens) known to have anthelmintic effect were added to the in vitro fermentation bottles containing the rumen fluid and medium. The rumen protozoal population was depressed by the addition of Pyrethrum, Torreya nucifera and Ruta graveolens. The methane production was also significantly (p<0.05)reduced by addition of Pyrethrum (2.20 ml/g DM), Torreya nucifera (2.36 ml/g DM) and Ruta graveolens (2.20 ml/g DM). The microbial growth in the treatments of Ruta graveolens or anthoxylum piperitum was the greatest after 12 h and 24 h incubations, respectively. The results of this study indicated that anthelmintic plant extracts appeared to reduce methane production by inhibition of ruminal protozoa related with the methanogens living endosymbiotic in protozoal cells.