• Journal of Animal Science and Technology
• /
• 제64권5호
• /
• pp.842-853
• /
• 2022

Ochratoxin A (OTA) is a well-known mycotoxin that causes disease through the ingestion of contaminated food or feed, for example, in the porcine industry. The intestinal epithelium acts as the first barrier against food contamination. We conducted a study on the exposure of the porcine intestinal epithelium to OTA. We used the intestinal porcine epithelial cell line IPEC-J2 as an in vitro model to evaluate the altered molecular mechanisms following OTA exposure. Gene expression profiling revealed that OTA upregulated 782 genes and downregulated 896, totalling 1678 differentially expressed genes. Furthermore, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting confirmed that OTA damages the tight junction protein ZO-1. Moreover, OTA activated the expression of inflammatory genes (IL-6, IL-8, IL-10, NF-kB, TLR4, and TNF-α). In summary, this study confirmed that OTA alters various molecular mechanisms and has several adverse effects on IPEC-J2 cells.

• Journal of Microbiology and Biotechnology
• /
• 제26권10호
• /
• pp.1687-1695
• /
• 2016

Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious threat to public health worldwide. Microbiological methods are known to be a promising approach for OTA biodegradation because physical and chemical methods have practical limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional Korean meju (a fermented starter for fermentation of soybeans) samples were examined for OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036 and M074 through sequence analysis of the beta-tubulin gene. After culturing both A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTA-biodegradation activity was analyzed using high-performance liquid chromatography (HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and 91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA (40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA production activity. Our data suggest that A. tubingensis isolates and their enzymes have the potential for practical application to reduce levels of OTA in food and feed.

• 미생물학회지
• /
• 제55권1호
• /
• pp.33-38
• /
• 2019

진균독소의 한 종류인 ochratoxin A (OTA)는 저장 곡물에 흔한 오염물질로 주로 Aspergillus에 의해 생성되어 인간과 가축의 건강을 위협한다. 이 연구의 목적은 분리 세균Bacillus subtilis AF13과 Streptomyces shenzhenensis YR226의 OTA 제거능을 조사하는 것이다. NB 배지에서 AF13과 YR226은 $100{\mu}g/L$ OTA를 24시간 동안 각각 94.23과 97.73% 제거하였다. 두 균주 배양액의 원심분리 후 분리된 세척 세포와 무세포 상등액의 OTA 제거능을 측정하였는데 두 균주의 상등액 모두 $100{\mu}g/L$ OTA를 수 초 내에 90% 이상 제거하였으며, YR226의 세척 세포도 OTA를 24시간에 98.88% 제거하였다. 배양 상등액의 고압멸균, proteinase K와 chymotrypsin 처리 시 그 OTA 제거능은 영향을 받지 않았으며 이는 두 균주들에 의해 분비된 열-안정성을 가진 비-단백질성 물질이 OTA 제거에 관련된 것을 추정할 수 있다. 이 결과는 AF13과 YR226 균주가 OTA로 오염된 곡물과 사료에서 OTA를 제거하는데 사용될 수 있으며 따라서 농업과 사료 산업에서 경제적 피해를 감소시킬 수 있을 것이다.

• 한국식품위생안전성학회지
• /
• 제22권4호
• /
• pp.353-358
• /
• 2007

국내에서 생산된 사료(2006-2007) 중 253점(배합사료 194, 단미사료 59)의 사료에서 오크라톡신의 오염도를 조사하였다. 단미사료 중 OTA오염도는 27%이었고 오염농도는 OTA 0.27-3.39 ppb 수준이었다. 배합사료에서 OTA 오염도가 76%로 나타났으며, 평균 0.21-13.64 ppb 수준의 검출농도로 분석되었다. 모든 사료 중 젖소사료(96%)> 닭사료(85%)> 돼지사료(79%) 순으로 OTA이 오염되어 있는 것으로 나타났으며, OTA평균오염농도는 고기소 사료(2.2 ppb)에서 가장 높았으며, 젖소사료(1.6 ppb), 박류(1.2 ppb)순으로 감소되는 것으로 나타났다.

• 약학회지
• /
• 제42권3호
• /
• pp.336-344
• /
• 1998

Effects of ochratoxin A (OTA) on embryo development were studied in cultured whole embryos from 9.5 day gestation rat for 48 h. OTA (more than $0.5{\mu}g/ml$) induced microcephaly in the cultured rat whole embryos. Protein and DNA content, and DNA synthesis were significantly inhibited by OTA. We next examined whether the microcephaly seen in cultured whole embryo partially results from inhibition of differentiation of embryonic midbrain cells. Embryonic midbrain cells were extracted from 12 day gestation rat embryos, and cultured for 96 hr. OTA ibhibited cell differentiation about 50% over control. We also tested whether OTA-induced embryotoxicity would be associated with oxidative damages. We measured the ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GT) and glutathione peroxidase (GPX) activities, and glutathione (GSH) content in both cultured whole embryos and embryonic midbrain cells. OTA decreased GSH content, whereas slightly increased ${\gamma}$-GT activity, but GPX activity was not significantly changed. These results show that OTA caused the microcephaly and its effect may be partially due to the inhibition of cell differentiation of embryonic midbrain cells, but the role of oxidative damages is not clear in embryotoxicity.

• Asian-Australasian Journal of Animal Sciences
• /
• 제25권1호
• /
• pp.114-121
• /
• 2012

The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

• Journal of Microbiology and Biotechnology
• /
• 제19권1호
• /
• pp.83-92
• /
• 2009

Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, respectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.

• 생명과학회지
• /
• 제16권6호
• /
• pp.1006-1013
• /
• 2006

식품안전에 대한 관심이 증가되고 있는 현재, 생물학적 화학적 위해요소로 분류되고 있고, 현재 많은 나라에서 규제치를 설정하고 있는 곰팡이 독소인 ochratoxin A(OTA)에 대한 정량적 측정이 가능한 고속검색법을 개발 하고자 단클론성 항체를 이용하여, 측정시 분리과정이 필요 없는 형광편광면역분석법(FPIA)을 개발하고 최적화 시켰다. 동일구조를 가지는 형광물질 표식자인 OTA-EDF를 합성하여 OTA에 대한 특이항체와 경쟁반응을 시켜 나타나는 형광-편광도(mP)의 변화를 측정하였다. 이는 면역분석법의 특이성과 민감성을 충분히 만족하였다. OTA의 검출범위는 0.5-200 ng/ml였고, 검출한계는 0.3 ng/ml였다. 개발된 분석법은 다른 곰팡이 독소들과의 교차반응은 없었고 높은 특이성과 재현성 및 회수율을 나타내었다. HPLC 방법에 의한 회수율은 88-84%로 다소낮게, FPlA법의 회수율은 90-110%로 다소 높게 나타났다. 16점의 현미시료를 분석하였을 때, 2점이 상관관계가 높게 12-20 ppb 정도 오염된 것으로 나타났다. 4점은 FPIA 및 HPLC 모두에서 음성으로 판정되었다. 개발된 FPIA는 복잡한 전처리 방법이 필요 없는 신속한 검색이 가능하므로, 식품 및 환경에서의 OTA 잔류 검사에 유용하게 사용될 수 있을 것이다.

• 동아시아식생활학회지
• /
• 제21권2호
• /
• pp.222-227
• /
• 2011

The Aspergillus sp. is well known as a fungus that causes black mold disease and secretes ochratoxin A (OTA). Our study found that infection by this fungus via inoculation onto grapes produced more severe symptom in wounded berries than in fresh berries. Furthermore, OTA contents were higher on the grape skins than in the fleshy portions of the grapes. OTA accumulated during the first 3 days after inoculation, but then gradually decreased. The OTA contents in wine made from 5 kg of grapes which included 400 g of infected grapes ranged from 0.17 to 0.37 ${\mu}g/mL$. An investigation of 25 marketed commercial wines showed the OTA contents were <1.2 ${\mu}g/mL$ which is lower than the limit of 2 ${\mu}g/mL$ established by the Korea Food & Drug Administration.

• 미생물학회지
• /
• 제55권3호
• /
• pp.226-233
• /
• 2019

Ochratoxin A (OTA)는 주로 Aspergillus가 생성하는 진균 독소의 하나로 저장 곡물의 흔한 오염물질로 인간과 가축의 건강을 위협하고 있다. 본 연구의 목적은 분리 세균인 Bacillus subtilis AF13과 Streptomyces shenzhenensis YR226의 OTA 생성 Aspergillus 균주들에 대한 생장과 OTA 생성 저해능 조사이다. OTA 생성 세 균주들의 균사 생장과 포자 형성에 대한 항진균활성은 한천 평판에서 AF13과 YR226 균주와의 공동배양에 의해 조사되었다. AF13과 YR226은 10일 배양 중 진균 집락 직경을 각각 77.58%와 78.48% 감소시켰으며, 두 균주 모두 포자 형성을 99%까지 저해하였다. YR226은 또한 세 진균 균주의 포자 발아도 91% 이상 감소시켰다. Yeast extract sucrose 배지에서 Aspergillus와 AF13 또는 YR226 균주를 동시배양하였을 때 세 종류 진균 모두 균사체 생장과 OTA 생성이 감소하였다. 특히 AF13은 A. alutaceus의 균사체 생장을 완전히 저해하고 OTA 생성은 99% 감소시켰으며, YR226은 균사체 생장과 독소 생성을 99%까지 저해하였다. AF13과 YR226이 생성하는 항진균물질에는 siderophore, chitinase, protease, ${\beta}$-1,3-glucanase와 biosurfactant가 포함된다. 이 결과는 AF13과 YR226이 독소생성 진균으로부터 가치있는 농작물을 보호하는 생물학적 방법으로 이용될 수 있으며, 따라서 농업과 사료산업에서 경제적 피해를 감소시킬 수 있을 것이다.