이 연구는 축산물 중 포레이트 및 대사산물 5종의 안전관리를 위한 공정분석법을 확립하기 위하여 수행하였으며 분석법의 선택성, 검출한계 및 정량한계, 회수율에 대한 검증을 통하여 포레이트 및 대사산물 5종의 공정시험법으로의 유효성을 확인하였다. 포레이트 및 대사산물 5종을 신속하고 효과적으로 동시에 분석하기 위하여 LC-MS/MS를 사용하였고, 1% 아세트산 포함 아세토니트릴 추출 후PSA, C18을 이용해 정제하였다. 개발된 분석법의 평균 회수율은 79.2-113.9%였으며, 분석오차는 19.2% 이하로 정확성 및 재현성이 우수함을 확인할 수 있었다. 개발된 분석법은 국제적 잔류농약 분석 가이드라인에 적합한 수준이었으며 축산물 중 포레이트 및 대사산물 5종의 잔류검사를 위한 공정분석법으로 활용할 수 있을 것으로 판단된다.

저온스트레스에 따른 차나무 잎의 변화와 카테킨, 카페인, 아미노산대사 그리고 환원당 등 다양한 대사물의 함량 변화를 구명하였다. 저온스트레스 피해를 입은 실외포장의 차나무 잎은 세포크기, 기공 및 엽맥세포의 크기가 저온스트레스를 입지 않은 온실 재배 녹차잎에 비해 작았다. 저온스트레스 피해를 입은 실외포장의 차나무 잎에서 카페인과 카테킨의 함량은 온실 재배 차나무보다 높았다. 그러나 아미노산의 함량은 저온스트레스 피해를 입은 실외포장의 차나무 잎보다 온실에서 재배한 차나무가 높았다. 환원당 함량은 실외포장의 차나무에서 낮았다. 인위적 저온스트레스 처리 결과 환원당의 함량은 저온스트레스 처리 2시간까지는 증가하였다가 이후는 감소하였다. 이 결과는 저온스트레스가 차나무 잎의 형태적 변화와 더불어 카페인과 카테킨 그리고 당 등 대사물의 영향을 미친다는 것을 나타낸다.

반추위 pH 조정제로서 일반적으로 중조가 많이 사용되고 있으나 사료에 중조의 첨가량을 증가시키는 경우에 기호성을 저하시키는 경향이 있다. 따라서 본 연구에서는 사료의 기호성에 영향을 미치지 않으면서 반추위 완충효과를 나타내도록 개발된 액상형 완충제의 젖소사료 첨가제로서의 이용효과를 구명하기 위하여 수행되었다. 시험축으로 착유 중기 젖소 27두를 공시하였으며, 처리는 대조구와 중조 첨가구, 그리고 액상형 완충제 첨가구로 하여 처리별 9두씩 배치하였다. 대조구는 착유우용 자가배합 TMR 사료를 급여하였으며, 중조 첨가구는 대조 사료에 중조를 두당 1일 150g 추가하여 급여하였고, 액상형 완충제 첨가구는 같은 방식으로 액상형 완충제를 두당 15㎖ 추가하여 사양시험을 수행하였다. 시험결과 산유량은 대조구, 중조 및 액상형 완충제 첨가구의 경우에 각각 23.82, 23.91, 24.91kg으로 대조구에 비하여 액상형 완충제 처리구에서 다소 증가하는 경향이었으나 처리 간에 뚜렷한 유의 있는 결과를 보여주지는 않았다. 처리에 따른 유성분 변화에서는 유단백질 및 무지고형분에서 액상형 완충제를 첨가한 경우에 대조구에 비해 유의 있는 증가(p < 0.05)를 보여주었고, 유지율에서도 대조구에 비하여 처리구가 다소 증가하는 경향을 보였다. 혈액성분 중 포도당과 콜레스테롤은 대조구에 비해 액상형 완충제의 첨가구에서 다소 증가하였으나, 처리 간에 유의 있는 차이를 나타내지는 않았다. 따라서 개발된 액상형 완충제의 추가급여 시 생산성 증대, 특히 유성분 증가 효과가 있어 반추위 pH 조정을 위하여 일반적으로 이용되고 있는 중조를 대체하여 반추동물의 사료첨가제로 이용이 가능할 것으로 평가되었다.

Deoxynivalenol (DON) and related trichothecene mycotoxins are extensively distributed in the cereal-based food and feed stuffs worldwide. Recent climate changes and global grain trade increased chance of exposure to more DON and related toxic metabolites in poorly managed production systems. Monitoring the biological and environmental exposures to the toxins are crucial in protecting human and animals from toxicities of the hazardous contaminants in food or feeds. Exposure biomarkers including urine DON itself are prone to shift to less harmful metabolites by intestinal microbiota and liver metabolic enzymes. De-epoxyfication of DON by gut microbes such as Eubacterium strain BBSH 797 and Eubacterium sp. DSM 11798 leads to more fecal secretion of DOM-1. By contrast, most of plant-derived DON-glucoside is also easily catabolized to free DON by gut microbes, which produces more burden to body. Phase 2 hepatic metabolism also contributes to the glucuronidation of DON, which can be useful urine biomarkers. However, chemical modification could be very typical depending on the anthropologic or genetic background, luminal bacteria, and hepatic metabolic enzyme susceptibility to the toxins in the diet. After toxin exposure, effect biomarkers are also important in estimating the linkage and mechanisms of foodborne diseases in human and animal population. Most prominent adverse effects are demonstrated in the DON-induced immunological and behavioral disorders. For instance, acutely elevated interleukin-8 from insulted gut exposed to dietaty DON is a dominant clinical biomarker in human and animals. Moreover, subchronic exposure to the toxins is associated with high levels of serum IgA, a biological mediator of IgA nephritis. In particular, anorexia monitoring using mouse models are recently developed to monitor the biological activities of DON-induced feed refusal. It is also mechanistically linked to alteration of serotoin and peptide YY, which are promising biomarkers of neurological disorders by the toxins. As animalalternative biomonitoring, huamn enterocyte-based assay has been developed and more realistic gut mimetic models would be useful in monitoring the effect biomarkers in resposne to toxic contaminants in the future investigations.

Rutin is one of the major flavonoids found in buckwheat (Fagopyrum esculentum Moench). While rutin is already known to exhibit anti-oxidative, anti-inflammatory, and anti-carcinogenic activities. However, the health beneficial function of rutin metabolites is not well understood. In DPPH radical scavenging assays, the present study found that 3,4-dihydroxyphenyl acetic acid had the highest total anti-oxidant activity, followed by 3,4-dihydroxyphenylacetic acid, rutin, homovanillic acid, and 3-hydroxyphenyl acetic acid. Further, 3,4-dihydroxyphenylacetic acid strongly reduced LPS-induced IL-6 production in RAW 264.7 cells, compared with other metabolites. Therefore, these results suggest that rutin metabolites have potential to be utilized as food ingredients with anti-oxidant and anti-inflammatory activities.

An experiment was conducted to study effects of abomasal infusions of casein, glucose or starch on milk production and blood metabolites in dairy cows. Four lactating cows (559±41.9kg) fitted with 100 mm ruminal cannulas were used in a 4 × 4 Latin square experiment. The cows were given access ad libitum to grass silage and received 6.2 kg dry matter (DM) of a mixture of rapeseed meal (5.3kg/d DM) and barley grain (0.9kg/d DM) as a basal diet (CON), the basal diet plus intra-abomasal infusions of 270 g sodium caseinate (CAS), 300 g of glucose (GLU) or 243 g of starch (STA) starting at 09:00 h each day. Abomasal infusion of different nutrients did not affect (p>0.05) grass silage intake and rumen fermentation parameters (rumen pH, ammonia N and volatile fatty acids). As unexpected, milk yield and composition data did not differ between CON and the treated groups (p>0.05). Most of plasma amino acids were not significantly (p>0.05) affected by the abomasal infusions of CAS, GLU or STA except for histidine, tryptophan and glycine. Although histidine, known as the first limiting amino acid in dairy cows fed grass silage based diets, was significantly (p=0.003) increased by CAS, we failed to result in a clear increase in milk yield and protein yield. This discrepancy between plasma AA profiles and milk production could be presumably explained by relatively high concentration of total amino acids in the present study (2,039μmol/l).

This study was carried out to investigate comparision anaylsis of metabolites in mandarin leave inoculated by Elsinoe fawcetii. For the analysis of metabolites, we performed using by UPLC-MS spectrometry in gradient eluent condition by acetonitrile and water. Flow rate was 0.4ml/min. Instruments conditions were followed by detection ion mode(positive ion), scan range(MS:m/z100-1000), spray voltage (4kV) ,capillary voltage(35v), capillary temp(300℃). Several flavones were detected in inoculated pathogen compared to control.

Entomopathogenic fungi are natural pathogens of insects and contribute to the regulation of host insect populations in the environment. Several these fungi produce a wide range of secreted enzymes, secreted protein toxins and secondary metabolites to overcome host defenses and ultimately kill the host, and to defend host resources against competing pathogens and saprophytes. This study was performed to evaluate the antimicrobial activity of 207 entomopathogenic fungi form Korea soils against plant pathogenic bacteria Ralstonia solanacearum and plant pathogenic fungi Botrytis cinerea using dual culture technique on SDYA. As results, twelve isolates (5.7%) and six isolates (2.8%) showing the greatest inhibition against R. solanacearum and B. cinerea, respectively. The culture supernatant of these selected isolates completely suppressed the growth of the pathogen, indicating that suppression was due to the presence of antimicrobial compound in the culture filtrate. The stability test of the culture filtrate showed that the antimicrobial component was heat stable and not protein. These entomopathogenic fungal metabolites may be a good feature to be used in the development of a new biocontrol method of R. solanacearum and B. cinerea.

An entomopathogenic bacterium, Xenorhabdus nematophila, secretes at least eight bacterial metabolites, which have suppressive effects on insect immunity. This study quantified their sequential production during bacterial growth and analyzed their individual immunosuppressive activities against an insect host, Spodoptera exigua. X. nematophila exhibited a typical bacterial growth in both insect host and culture medium, in which eight metabolites were secreted in different time points. At early growth phase (6 to 12 h), Ac-FGV, Cis-cPY, PHPP and indole metabolites were detected in the culture broth. During early growth phase, PHPP was highly potent to inhibit phenoloxidase activity as well as nodule formation. At late growth phase (24 to 48 h), BZA, HPA, PY were detected at 10 – 140 ppm in the culture broth, their metabolites were highly potent to inhibit phospholipase A2 and to induce cytotoxicity to hemocytes. These results suggest that X. nematophila sequentially produces the immune suppressive metabolites, which cooperatively inhibit different steps of insect immune responses.

An entomopathogenic bacteria, Xenorhabdus nematophila (Xn) and Photorhabdus temperata subsp temperata (Ptt), suppresses insect immune responses and facilitates its symbiotic nematode development in target insect. Benzylideneacetone (BZA), PY, cPY, Ac-FGV, indole, 2-oxindole and 3-(4-hydroxyphenylpropionic) acid (PHPP) were compounds derived from the bacterial. Their immunosuppressive activities have been induced by inhibitory activity against eicosanoid biosynthesis and used to develop an additive to enhance control efficacy of other commercial microbial insecticides. This study investigated any cytotoxicity of their culture broth and bacterial metabolites on Spodoptera exigua hemocyte. When Xn or Ptt (<100 cells per larva) were injected to larval of S. exigua, the bacteria increased in density with incubation time, while the insent hemocyte numbers significantly and the resulting culture broths were sampled for analysis of their cytotoxicity against S. exigua hemocytes. In addition, the sequential culture broth samples were analyzed in active component chemicals using a reverse phase HPLC. Finally, seven bacterial metabolites were analyzed in relative cytotoxicity against S. exigua. These results suggest that BZA is a major cytotoxic compound.

본 시험은 포도부산물의 첨가가 젖소의 산유량, 유성분 및 혈액 성상에 미치는 영향을 조사하기 위하여 실시하였다. 비유중기 젖소 8두(평균 월령: 89.8, 평균 유기: 164.4, 평균 산차: 2.8산)를 공시하였으며, 포도부산물을 첨가하지 않은 대조구와 1일 1 kg을 첨가한 시험구로 두 처리구를 설정하여 3주간 급여하였다. 유지방, 유단백, 유당 및 총 고형물 함량은 처리간에 차이가 없었다. 그러나 산유량은 포도부산물 을 첨가한 처리구가 23.13 kg/d으로 대조구의 21.16 kg/d에 비해 높은 반면(p<0.05), 체세포 수는 대조구가 포도부산물 처리구에 비해 높았다(p<0.05). 혈중 glucose, alanine aminotransferase, total protein 및 blood urea nitrogen 함량은 처리구간 차이가 없었으나, AST와 총콜레스테롤 함량은 대조구 (123.75 IU/L, 200.82 mg/dl)가 포도부산물 처리구(95.34 IU/L, 180.63 mg/dl)에 비해 높았다(p<0.05). 이상의 결과에서, 다양한 생리활성을 가진 포도부산물은 젖소를 위한 기능성 사료로 이용 가능성이 높을 것으로 사료된다.

Bacillus thuringiensis (Bt) is a bacterial biopesticide against insect pests, mainly lepidopterans. Spodoptera exigua and Plutella xylostella exhibit significant decreases in Bt susceptibility in late larval instars. To enhance Bt pathogenicity, we used a mixture treatment of Bt and other bacterial metabolites which possessed significant immunosuppressive activities. Mixtures of Bt with culture broths of Xenorhabdus nematophila (Xn) or Photorhabdus temperata ssp. temperata (Ptt) significantly enhanced the Bt pathogenicity against late larval instars. Different ratios of Bt to bacterial culture broth had significant pathogenicities against last instar P. xylostella and S. exigua. Five compounds identified from the bacterial culture broth also enhanced Bt pathogenicity. After determining the optimal ratios, the mixture was applied to cabbage infested by late ins tar P. xylostella or S. exigua in greenhouse conditions. A mixture of Bt and Xn culture broth killed 100% of both insect pests when it was sprayed twice, while Bt alone killed less than 80% or 60% of P. xylostella and S. exigua, respectively. Other Bt mixtures, including Ptt culture broth or bacterial metabolites, also significantly increased pathogenicity in the semi-field assays. These results demonstrated that the Bt mixtures collectively names 'Bt-Plus' can be developed into potent biopesticides to increase the efficacy of Bt.

본 연구는 반추위 cannula를 장착한 거세한우 4두(평균체중 600 48.4 kg)를 공시하여 수확시기별(유숙기, 호숙기, 황숙기 및 완숙기) 총체벼 사일리지의 급여가 한우 반추위 발효패턴 및 혈액대사물질 변화에 미치는 영향에 대해서 조사하여 총체벼 가축사료화를 위해 최적의 수확기를 제시코자 실시하였다. 시간대별 채취한 반추위 소화액의 평균 pH는 처리구 간 통계적 유의성이 나타나지 않았다(p>0.05). 일일 반추위 pH 패턴에서 완숙기 처리구의

Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.