Allicin (AL) is one of the biologically active substance in garlic. Many researchers found that AL exhibits strong antioxidant activity and considered to represent anti-aging effect in vitro. The objective of this study was to investigate the effects of allicin treatment during porcine oocyte aging and their in vitro development. The oocyte was maturated in vitro for 44 h (control) without AL or 44+24 h IVM (24 h aging) with 0, 0.1, 1, 10 and 100 μM allicin (0 AL, 0.1 AL, 1 AL, 10 AL and 100 AL). The 1 AL treated group was significantly increased on maturation rate compared to the 0 AL treated groups, but the other treated groups were not different compared to the 0 AL treated group (p < 0.05). The 1 AL treated group was significantly increased on normal spindle formation and chromosome alignment compared to 0 AL treated group. We checked the effect of AL on parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development using aged oocytes. PA embryos in 1 AL treated group significantly increased the cleavage and blastocyst formation rate compared to control (p < 0.05). However, SCNT embryos of 1 AL treated group were no significant differences in embryo development. In PA embryo quality, the total cell number was significantly higher in 0.1 AL, 1 AL and 10 AL treated groups than control and DNA fragmentation rate in 1 AL was the lowest among all groups. SCNT embryo in 1 AL significantly increased total cell number and decreased DNA fragmentation. The AL treatment on aged oocyte enhanced PA and SCNT embryo developmental capacity. Therefore, AL may be helpful for assisted reproductive technology applying aged oocytes.

Somatic cell nuclear transfer (SCNT) technique is a key point of producing transgenic animal disease models. During in vitro production of SCNT embryo, the quality of matured oocytes are one of the important factors that regulate embryo developmental capacity. In preliminary test, we confirmed the effect of fibroblast growth factor 10 (FGF10) on porcine oocyte maturation. In this study, we investigated the developmental potential of SCNT embryos treated with the 10 ng/ml FGF10 (10 F) during in vitro maturation of recipient oocytes. The polar body emission rate was significantly higher in the 10 F treated group than control group. After SCNT, although the rate of fusion was no significant difference, the rate of cleavage and blastocyst formation was significantly increased in the 10 F treated group (p<0.05). In 10 F treated group, the total cell number was increased and the percentage of apoptotic cell was decreased in the blastocyst stage at day 7 (p<0.1). The transcription level of apoptosis relative gene, Casp3 was significantly decreased, while anti-apoptosis gene BCL2l1 was increased in the 10 F treated group compared to control group. The 10 F treated group was highly expressed the reprogramming related genes, Sox2 and POU5f1. Also, the first cleaving time was more faster and the percentage of cell block was significantly lower in 10 F treated group than in control group. In this study, we confirmed that 10 ng/ml FGF10 has effect on enhance the oocyte maturation and developmental capacity. These results demonstrate that FGF10 treatment can be used for in vitro development of porcine SCNT embryos and subsequent production of transgenic animal model.

Increase of bovine embryos produced by in vitro fertilization (IVF) has been seen. The main reason for producing in vitro fertilized embryos in Korea has been to utilize the genetics of cows with higher carcass grade. Ovaries are collected from the cows in the slaughter house and the information on the carcass grade of the cow can be traced. Embryos produced from cows with higher carcass grade have been favored by the farmers. PCR has been one of the main techniques for sex determination of embryos targeting various genes. Bovine sex determining region Y (SRY) is specific to Y chromosome. However, it requires a control gene for PCR, if the embryo is female. In comparison to SRY, amelogenin can be amplified from male or female embryos with different fragment sizes due to differential splicing in all bovidae. The goal of this study was to determine whether there are any differences in the sex ratio of embryos produced in vitro and to compare the efficiency of sex determination using PCR. Ovaries of Hanwoo were collected and transported to the laboratory in thermal bottles. For in vitro maturation, oocytes were collected from the follicles with less than 8 mm of diameter and placed in either the Brackett & Oliphant media (BO), Tissue culture medium-199 (TCM-199), or IVMD101 media, containing 3% fetal bovine serum (FBS), 0.5 mg/ml FSH, 0.5 mg/ml LH, and 1 mg/ml estradiol-17β. For IVF, frozen sperm from Hanwoo bulls were used. After 22-24h IVF, embryos were transferred and cultured either in BO or TCM-199 with 10% FBS until the embryos were hatched. Hatched blastocysts were stored in PBS frozen, and later thawed and treated with embryo lysis buffer. After isolating genomic DNA, it was used for PCR using primers for casein beta (CSN2), as PCR control, or for male specific SRY primers. Alternatively, primers for amelogenin were used. Sex of embryos was determined and the sex ratio was analyzed. Out of 94 embryos, sex of 83 embryos (88.3%) was determined and there were 40 male embryos (48.2%) and 43 female embryos (51.8%). Sex of 31 embryos was determined using both SRY and amelogenin. Among those, 17 embryos were determined as having identical sex, while 1 embryo was determined as having different sex, and the sex of 11 and 2 embryos were determined only by amelogenin or SRY primers, respectively. In conclusion, the success of determining the sex of embryos by PCR was relatively high. Using amelogenin primer for PCR tends to be more efficient than SRY primer in determining the sex. Slightly higher ratio of female embryos was different from previous years and the cause for the difference may require further investigation.

In vitro production of mammalian embryos has been achieved with the oocytes derived from middle-size follicles (MF, mainly 3-6 mm in diameter) in many species including domestic animals. In the ovaries, however, there are more small-size follicles with less than 3 mm in diameter (SF). If we can develop an efficient system to produce embryos in vitro from the oocytes from SF. In this presentation, I would like to review about embryo production in vitro from the oocytes derived from SF. As well as the diameter of oocytes, the number of cumulus cells surrounding the oocyte derived from SF is significantly smaller those of oocytes from MF. The comparative analysis in electrophoresis about secretions of cumulus-oocyte complexes derived from SF and MF demonstrated a significant difference in the proteins with a molecular weight. Proteins secreted from cumulus cells, vascular endothelial growth factor (VEGF), are 34- to 42-kDa proteins, including seven family members. The molecular weight of VEGF was similar with the secretion we observed. Supplementation of medium for in vitro maturation with VEGF significantly improved the oocytes competence not only to complete the meiosis in vitro but also to develop to the blastocyst stage following parthenogenetical activation. Removing cumulus cells 20 h after the start of culture for in vitro maturation also significantly improved the competence of oocytes derived from SF to achieve the meiosis. A combination of these new techniques may improve more the meiotic and developmental competences.

금(Au)나노물질은 광학 및 바이오 분야에서 다양하게 응용될 수 있을 뿐만 아니라 식품 농작물 재배에 무기비료제 로 사용 시 기능성 및 품질 향상에 긍정적인 영향을 줄 수 있는 것으로 알려지면서 금 나노입자를 이용한 건강기능식 품 또는 기능성 농작물 재배에 대한 연구개발이 진행되고 있다. 따라서 식품 농작물 잔류와 체내 흡수 가능성에 따른 식품 및 생체 내에서의 정확하고 정밀한 분석법 확립과 안전성검증이 요구된다. 본 연구에서는 유도결합플라즈마 질 량분석기(ICP-MS)를 이용한 금 나노입자의 정량 분석을 확립하고자 전처리 방법을 최적화하고, 회수율, 정확성 및 정 밀성을 분석하여Au의 분석법을 확립하였다. 확립된 Au 정량 분석법을 이용하여 생체시료 내에서 금 나노입자의 정량 분석을 수행하여 생체 매트릭스의 영향을 확인하였다. 이러한 분석법을 바탕으로 장내 micro fold (M) 세포를 모사한 follicle associated epithelium (FAE) 모델과 일반 장관 상피세포 조건을 모사한 Caco-2 monolayer 모델을 이용하여 금 나노입자의 유입량을 비교 연구하였다. 그 결과, 생체 매트릭스 존재에 따른 금 나노입자 정량분석에 미치는 영향은 없는 것으로 나타났으며, 금 나노입자는 장관 상피세포 내 M 세포에 의해 에너지 의존적 endocytosis 기작에 의해 체 내 유입될 수 있는 것으로 확인되었다. 이러한 식품 농작물 무기비료로서 금 나노입자의 식품 및 체내 분석법 확립연 구 및 체내 흡수기작 규명은 추후 안전성 평가에 기초자료로 활용될 수 있을 것이다.

(-)-Epicatechin gallate (ECG) is a polyphenol compound of green tea exhibiting biological activities, such as antioxidant and anticancer effects. To examine the effect of ECG on porcine oocytes during in vitro maturation (IVM), oocytes were treated with 0-, 5-, 15-, and 25 μM ECG. After maturation, we investigated nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels and subsequent embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF). After 42 hours of IVM, the 5 μM group exhibited significantly increased (p< 0.05) nuclear maturation (89.8%) compared with the control group (86.1%). However, the 25 μM group observed significantly decreased (p< 0.05) nuclear maturation (83.5%). In intracellular maturation assessment the 5-, 15-, and 25 μM groups had significantly increased (p< 0.05) GSH levels and decreased ROS levels compared with the controls. The 5- and 15 μM group showed significantly increased (p< 0.05) embryo formation rates and total cell number of blastocysts after PA (18% and 68.9, 15% and 85.1 vs. 12% and 59.5, respectively) compared with controls. Although the 25 μM group observed significantly lower blastocyst formation rates after PA (27.6% vs. 23.2%) than control group, the 5 μM group showed significantly increased blastocyst formation rates after PA (37.2% vs. 23.2%) compared to the control group. Furthermore, the 5 μM group measured significantly increased blastocyst formation rates (20.7% vs. 8.6%) and total cell number after IVF (88.3±1.5 vs. 58.0±3.6) compared to the control group. The treatment of 5 μM ECG during IVM affectively improved the porcine embryonic developmental competence by regulating intracellular oxidative stress during IVM.

This study was conducted to examine the effects of medium composition on organogenesis towards in-vitro cultured diploid and tetraploid Codonopsis lanceolata and obtain in-vitro mass propagation of superior species of C. lanceolata. Regarding MS medium composition for each concentration, diploid C. lanceolata was found to be declined. However, shoot and adventitious root formation were suppressed with higher mineral salt concentration, and active growth of shootand adventitious root was exhibited as 4.9 cm and 3.2 cm respectively in 1/2 MS medium. While in tetraploid C. lanceolata, it showed 2.9 cm and 3.2 cm respectively in 1/4 MS medium. In the case of sucrose concentration, no consistent decrease was observed for growth of shoot and adventitious root of diploid both at high and low concentration. The growth of shoot (at 3% concentration) and adventitious root (at 7% concentration) was 2.3 cm and 2.0 cm respectively. Although there was no difference in shoot formation of tetraploid C. lanceolata in all concentrations with the range of 1.7 ~ 1.8, there was a slight decrease in shoot growth at high concentration. Results revealed that the adventitious root formation was suppressed at high concentration. Concentration of agar exhibited no significant difference in shoot formation of diploid C. lanceolata at all concentrations. The highest result of adventitious growth (4.1 cm) was observed at 0.8% concentration. Slight inhibition of shoot formation and root formation of tetraploid C. lanceolata was observed at higher concentration. Shoot formation of diploid C. lanceolata also exhibited inhibition at higher concentration. Shoot formation of diploid C. lanceolata was increased at lower pH and shoot growth was the highest (2.3 cm) at pH 3.8. Adventitious root formation was higher at lower pH. Although there was no difference in shoot formation of tetraploid C. lanceolata presenting 1.7 ~ 1.8 regardless of high and low pH, growth inhibition was showed at higher pH. Adventitious root formation and growth showed a little higher result at pH 5.8.

The study aims to assess the embryo development and survivability of bovine embryos cultured in vitro by addition of cysteine. The rates of metaphase II formation are not significantly different among the three groups(73.8% for TCM199, 76.9% for TCM199 with 0.3mM cysteine and 83.8% for TCM199 with 0.5mM cysteine). No differences on cleavage rate(70.6~74.6%) was observed among three culture medium(70.6% for TCM199, 71.3% for CR1aa, and 74.6% for SOF) with 0.5mM cysteine. However, significantly(P<0.05) higher development rate was obtained in the blastocyst stage by adding 0.5mM cysteine in SOF medium(35.6%) than in TCM199(27.6%) or CR1aa(26.6%). No significant differences in the cleavage rates were observed among the three culture. After freezing the blastocysts cultured with 0.5M cysteine, the re-expansion rates ranged from 61.3% to 86.4% among groups, and hatching rates are from 26.3% to 46.9% among groups. The rates of re-expansion and hatching are significantly(P<0.05) higher in SOF medium(86.4% and 46.9%, respectively) than those in TCM199(61.3% and 26.3%) and CR1aa medium(87.1 and 44.4%). After thawing, the blastocyst re-expansion rate become significantly(P<0.05) higher in in vivo (87.1%) and in vitro (70.3%) embryos. In conclusion, our results demonstrate that supplementation of IVM and IVC media with 0.5mM cysteine improved the quality of in vitro production of embryo and post-thawed embryo. Future studies comparing these culture systems in well-designed trials should be performed.

Historically, Korea old cattle had been consisted with various lines of coat color brindle, black and white-brown breeds or more. The two rare lines of black and white coat color are maintained for animal resources and preserved critically. The present study was carried out to evaluate potential usage of cysteamine supplementation during in vitro matration (IVM) and in vitro culture/production of embryo (IVP) by transvaginal ultrasound-guided follicle aspiration (Ovum Pick-Up: OPU) for the establishment of cryo-banking system. Immature slaughterhouse-derived cumulus-oocyte complexes (SL-COCs) were matured in IVM medium supplemented with 0, 0.1, 0.3 or 0.9 mM cysteamine, and then cultured in mSOF-BAS for 8 days after in vitro fertilization. The treatment of 0.1 mM cysteamine on SL-COCs showed higher rate of blastocyst, so OPU-derived COCs from rare breeds were matured in TCM media supplemented with or without 0.1 mM cysteamine, FSH and 5% FBS. The embryos were evaluated their developmental stages on day 8. During IVM, cysteamine treatment significantly increased the embryo production rate of slaughterhouse-derived COCs (19.6% vs. 30.5%). The presence of cysteamine during IVM of OPU-derived COCs from rare Korean cattle breeds (albino white and black line) also increased embryo production rates than those from SL-COCs (27.4% vs. 41.9% and 36.4%). With these results, cysteamine treatment during IVM is one of key factors IVP of blastocysts to establish banking system of endangered rare Koarean cattle with OPU derived transferable blastocysts.

During the ovary preservation in low temperature, the cumulus oocyte complexes(COCs) lose their developmental competences after in vitro fertilization. We used phosphate-buffered saline (PBS) as a basic solutions of at various temperatures (25, 15 or 5 ℃) and supplemented them with 1mM glucose and 0.5mM glutamine as a source of carbohydrate metabolites. After recovery of COCs and in vitro fertilization, a significantly higher number of oocytes developed into blastocysts. The developmental competence of embryos that were originated from ovaries preserved at 15 ℃ was increased compared to those of 25 or 5 ℃. The maturation rate of oocytes was not differed between 24 and 36 h at 15 ℃ but showed lower than control group (71% versus 78%). In vitro-fertilized oocytes from ovaries stored at 25 ℃ for 24 h or at 5 ℃ for 24 h had a significantly decreased developmental potentials, but at 15 ℃ did not (27% versus 29% of blastocysts to develop into day 8). With these results, bovine ovaries can be preserved at 15 ℃ for 36 h without decreasing developmental capacity of in vitro-fertilized oocyte at least to the blastocyst stage. This information provides valuable information of preserving ovaries for embryo transfer or in vitro embryo production.

This study analyzed the antioxidant properties of Equisetum arvense and its effects on serum factor levels in mice fed a high-fat diet. The aim was to establish a new effective resource for biologically active materials. E. arvense stem and root extracts were obtained using deionized water at 95℃, and 70.5% ethanol at 85℃. These extracts were used to analyze the total phenolic compounds and antioxidant (ABTS, DDPH, and FRAP) activities. The effects of prepared ground samples were evaluated by feeding them to mice. E. arvense extracts showed strong antioxidant effects. The caffeic acid content was highest in the 70.5% ethanol extract of the vegetative stem, as determined by high-performance liquid chromatography. The blood concentrations of insulin and leptin were significantly lower in mice fed a high-fat diet supplemented with extracts of the root, reproductive stem, and vegetative stem of E. arvense than in mice fed only a high-fat diet. These results suggest that the polyphenolic compounds in E. arvense extracts exert various antioxidant effects. The stems and root of E. arvense can lower the blood levels of insulin and leptin, even after consumption of a high-fat diet.

Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at 38.5 ˚C under 5% CO2 in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at 118.3 ± 2.5 days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

Carbon source, an essential nutrient for plant growth, mainly includes exogenous sugar and CO2 of the environment in vitro. Therefore, the exogenous sugar and CO2 of the environment make the important roles in tissue culture. The aim of this study is to investigate the effects of different sugar concentrations (0, 10, 15 and 30 g·L-1) on the growth of colored Zantedeschia in vitro under certain CO2 concentration and explore the optimal sugar concentration. The plantlets in vitro of colored Zantedeschia had the largest root number, root weight, and root vigor under 0 g·L-1 (sugar-free culture) treatment. And they had the largest plant height, leaf length and leaf chlorophyll content, but p oor r oot v igor under 3 0 g·L-1 sugar. This study indicated that the optimal condition for proliferation and seedling culture of colored Zantedeschia plantlets in vitro was MS medium with 30 g·L-1 sugar, and the suitable medium for rooting culture and transplanting of colored Zantedeschia was MS medium with sugar-free culture under CO2 enrichment condition.

In the present study, the plantlets in vitro of Paeonia suffruticosa ‘Wu Long Peng Sheng’ were used as laboratory materials. The proteome during adventitious root induction process was investigated to sift the related proteins by two-dimensional electrophoresis and mass spectrometry. The results indicated that the protein spots were concentrated in the acidity gel region (pH 4 - 7) and the spots number had a dynamic change ranged from 373 to 462 at the process of root induction (0 – 7 d). 8 spots significantly changed were analyzed with a mass spectrometer and identified using associated software and databases. The peptide information of the 8 spots was similar to the ATP synthase β-subunit of P. suffruticosa (Spots 1 - 4 and 8), P. tenuifolia (Spots 5), P. californica (Spot 6) and P. brownie ( Spots 7) r espectiv ely. T he expression levels of protein spots 1, 4, 5, 6 and 7 was dramatically downregulated, and that of protein spots 2 and 3 had a slightly opposite tendency on the 3rd day. The obviously decreased period is particularly interesting as it was consistent with the induction period of adventitious root primordial of tree peony plantlet in vitro. The ATP synthase β-subunit could be consumed for assembling the ATP synthase in order to supply energy to the rooting process. Therefore, we speculated that the ATP synthase β-subunit was involved in adventitious root initiation of tree peony plantlets in vitro and we expect that further studies should be carried out in order to export its action mechanism.

This study was conducted to evaluate effects of supplementation of Bifidobacterium ruminantium on in vitro ruminal fermentation and methane production. Ruminal fermentation characteristics of Timothy hay (C1), whole barley (C2), Timothy hay + B. ruminantium (T1) and whole barley + B. ruminantium (T2) were evaluated at 0, 3, 6, 9, 12, 24 and 48 h incubation at 39℃. The amount of B. ruminantium culture added into T1 and T2 was 0.3 ml. The pH values ranged from 5.99 to 6.83 in all the treatments. Concentration of NH3-N of C2 and T2 was higher than C1 and T1 at 48 h (p<0.05). The total gas production of C2 and T2 was higher than C1 and T1 at 9 h (p<0.05). The total methane production of treatments with B. ruminantium was not significantly different at 24 and 48 h (p<0.05). Concentration of lactic acid was significantly different between both substrates (p<0.05). Therefore, B. ruminantium supplementation was determined to be insignificant in the in vitro ruminal fermentation and methane production, while a further study was required to investigate relation to lactic acid production with different forage sources.

강원도에서 육성한 유색칼라 ‘Lip Glow’와 외국품종인 ‘Captain Safari’ 조직배양묘의 최적 순화 기술 개발로 종구 자 급화 및 농가 실질 소득 증대에 기여하고자 본 연구를 실시하 였다. 순화실은 하우스 형태(1.2 × 6 × 1.5m)이며, 차광(0, 50%)과 흑색차광망(50, 90%) + PE 필름으로 밀폐한 것을 비 교하였다. ‘Lip Glow’의 순화는 흑색차광망 90% + PE 필름 처 리에서 23일째부터 초장이 가장 크고 엽수가 많았고 이후 생 육속도도 빨랐다. 정식 후 45일째의 근수가 많고 근장도 길었 으며, 생체중은 무차광의 1.6배, 건조중은 1.4배 무거웠고, 괴 경 생존율은 81.1%로 무차광 34.2%에 비해 우수하였다. 유색 칼라 ‘Captain Safari’의 순화는 흑색차광망 50% + PE 필름과 흑색차광망 90% + PE 필름 처리에서 23일과 30일째 초장이 크고 엽수가 많았고 이후 생육속도도 빨랐다. 정식 후 45일째 흑색차광망 50% + PE 필름 처리의 초장이 가장 크고 및 엽장 도 가장 많았다. 정식 후 61일째 차광 50% + PE 필름 처리에 서 근수 8.9개로 가장 많고, 근장 13.1cm로 가장 길고, 생체중 5.6g로 무차광의 1.9배, 건조중 1.0g로 무차광의 2.6배 더 무 거웠다. 수확 후 괴경중은 차광 50% + PE 필름 처리에서 5.6g으로 가장 무겁고, 구주도 6.7cm로 가장 컸으며, 생존율 60.6%로 가장 높았지만 차광 50%와 차광 90% + PE 필름 처 리와 유의차는 없었다. 따라서 유색칼라 조직배양묘의 최적 순화 기술로 국내품종인 ‘Lip Glow’는 차광 90% + PE 필름 처 리가, 외국품종인 ‘Captain Safari’ 는 PE 필름 여부와 관계없 이 차광 50% 이상 처리가 효과적이었다.

본 시험은 질산염함유 식물 추출물을 이용하여 in vitro 반추위 발효성상 및 반추위 메탄 발생에 미 치는 영향을 알아보기 위해 수행되었다. 공시재료는 감자, 당근, 배추, 상추 및 시금치 추출물을 사용하 였으며, 발효 기질은 2mm로 분쇄된 timothy 0.3g을 사용하였다. In vitro 실험은 조사료(timothy) 및 배합사료를 6:4의 비율로 급여한 반추위 cannnula가 시술된 한우 암소에서 채취하였다. 반추위액과 McDougall’s buffer를 1:2의 비율로 혼합한 발효액을 0.3g timothy와 식물 추출물이 담긴 50mL serum bottle에 혐기상태로 20mL 분주한 뒤, 39℃에서 9, 12, 24, 48시간 동안 발효하였다. pH는 발 효 시간이 증가함에 따라 점점 감소하며, 6.31~6.96으로 정상범위였다. 건물 소화율은 9시간 배추 추 출물구에서 유의적으로 낮았고, 암모니아 농도는 감자, 배추, 상추 추출물구에서 대조구에 비해 첨가구 에서 유의적으로(p<0.05) 낮았으며, 반추위 미생물 성장량은 발효 24시간대의 당근 추출물구에서 높았 다(p<0.05). Acetate와 propionate 농도는 대조구에 비해 첨가구에서 유의적으로 낮았으며(p<0.05) butyrate 농도는 첨가구간에 발효시간별로 각각 다른 양상을 보였다. 총 가스 발생량은 발효 12시간 및 24시간대에서 배추, 상추, 시금치 추출물구에서 유의적으로(p<0.05) 낮았으며, 메탄 발생량은 감자, 배 추, 시금치 추출물구가 대조구에 비해 유의적으로(p<0.05) 낮았고, 이산화탄소 발생량 또한 첨가구에 비해 대조구에서 유의적으로(p<0.05) 낮았다. 결과적으로 본 시험에 사용된 5종의 질산염 함유 식물 추 출물은 반추위 발효에 악영향을 미치지 않으면서 메탄저감 효과를 나타내었다. 특히 배추 추출물은 반 추위내 메탄발생을 저감시킬 수 있는 식물추출물로서 가능성이 높은 것으로 사료된다.