Background: Aflatoxin B1 (AFB1) is a toxic metabolite generated by Aspergillus species and is commonly detected during the processing and storage of food; it is considered a group I carcinogen. The hepatotoxic effects, diseases, and mechanisms induced by AFB1 owing to chronic or acute exposure are well documented; however, there is a lack of research on its effects on the intestine, which is a crucial organ in the digestive process. Dogs are often susceptible to chronic AFB1 exposure owing to lack of variation in their diet, unlike humans, thereby rendering them prone to its effects. Therefore, we investigated the effects of AFB1 on canine small intestinal epithelial primary cells (CSIc). Methods: We treated CSIc with various concentrations of AFB1 (0, 1.25, 2.5, 5, 10, 20, 40, and 80 μM) for 24 h and analyzed cell viability and transepithelial-transendothelial electrical resistance (TEER) value. Additionally, we analyzed the mRNA expression of tight junction-related genes (OCLN, CLDN3, TJP1, and MUC2), antioxidant-related genes (CAT and GPX1), and apoptosis-related genes (BCL2, Bax, and TP53). Results: We found a significant decrease in CSIc viability and TEER values after treatment with AFB1 at concentrations of 20 μM or higher. Quantitative polymerase chain reaction analysis indicated a downregulation of OCLN, CLDN3, and TJP1 in CSIc treated with 20 μM or higher concentrations of AFB1. Additionally, AFB1 treatment downregulated CAT , GPX1, and BCL2. Conclusions: Acute exposure of CSIc to AFB1 induces toxicity, and exposure to AFB1 above a certain threshold compromises the barrier integrity of CSIc.

메주와 누룩은 한국 전통 발효 식품에 사용되는 스타터로, Aspergillus속 곰팡이나 aflatoxin에 노출되기 쉽다. 본 연구에서는 우리나라에서 시판되는 57개의 메주 시료와 18개의 누룩 시료로부터 Aspergillus 속 곰팡이를 분리하고 동정하였다. 분리주의 aflatoxin 생성 가능성을 평가하기 위하여 multiplex PCR을 통해 aflatoxin 생합성 유전자 (aflO, aflP, aflR)를 확인하고, 이들 분리주에 의해 생성되는 aflatoxin 함량을 HPLC로 조사하였다. 뿐만 아니라 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석하였다. 그 결과, 메주 시료로부터 130개, 누룩 시료로부터 47개 균주가 분리되어 총 177개의 분리주를 확인 및 동정하였다. 각 각 메주와 누룩으로부터 분리된 19.2% (25/130), 10.6% (5/ 47)의 분리주가 3 종류의 aflatoxin 생합성 유전자를 모두 보유하였으며, 그 중 메주로부터 분리된 5개의 분리주가 실제 로 aflatoxin을 생성하였다. 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석한 결과, 88% (51/58)의 메주 시료의 aflatoxin 오염 수준은 모두 검출한계 미만으로 나타났고, 누룩 또한 시료의 39% (7/18)가 검출한계 미만으로 확인되었다. 메주와 누룩에서 분리된 분리주 중 aflatoxin 생합성 유전자를 모두 보유하거나 배지 상에서 aflatoxin 생성을 보여준 aflatoxigenic 균주는 존재하였으나 유통되고 있는 시료에서 aflatoxin 오염 빈도는 낮은 수준임을 확인할 수 있었다.

In the current study, 109 commercial nut samples were collected from different Korean markets and analyzed for the contamination of 5 different mycotoxins (aflatoxin, ochratoxin A, deoxynivalenol, zearalenone, and T-2 toxin) using ELISA kits. The results revealed that the most frequently detected mycotoxin was zearalenone (n=36, 33%), followed by aflatoxin (n=31, 28.4%) and ochratoxin A (n=30, 27.5%). Deoxynivalenol and T-2 toxin were also detected in 22 (20.3%) samples, respectively. Among 109 nut samples, 33 samples (30.3%) were contaminated only with one kind of mycotoxin, whereas 43 samples had at least 2 kinds of mycotoxins. Two samples were contaminated with as many as 4 different mycotoxins, and they were both walnuts. Although the monitoring results revealed the amount of aflatoxin contamination was under the safety criteria, there is no current safety guideline for other kinds of mycotoxins or multiple contaminations in Korea. Therefore, further studies should be performed to reveal the distribution of mycotoxin in different foods and propose appropriate safety guidelines for Korean markets.

Mycotoxins, such as aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisin B1 (FMB1), ochratoxin A, T2 toxin, and zearalenone, are found in numerous vegetables. Mycotoxin accumulation in food and feed poses serious health risks to humans and animals because of carcinogenic, mutagenic, teratogenic, and toxic properties. In addition, mycotoxins cause large economic losses in commercial crop production, food and feed processing, and animal husbandry worldwide. In this study, an analytical method for the simultaneous analysis of the levels of AFB1, DON, and FMB1 in cow blood with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated. AOZTM and Myco6in1TM multitoxin immunoaffinity columns and an OasisTM reversed-phase solid-phase extraction Hydrophilic-Lipophilic-Balanced columns were used to purify and concentrate the blood samples. Extracts that contained AFB1, DON, and FMB1 had average recovery of 64.0%, 98.0%, and 89.9%, respectively. In conclusion, we used LC-MS/MS to detect several important toxicological mycotoxins in cow blood. The multimycotoxin method, which detected and quantified the levels of AFB1, DON, and FMB1 can be used in animal pilot studies to monitor simultaneous exposure to major mycotoxins.

본 연구는 국내산 곶감의 미생물 및 aflatoxin에 대한 안전성을 평가하여 곶감의 안전관리의 기초자료로 활용하고자 수행하였다. 본 연구를 위하여 반건시 34농가, 건시 61 농가, 감말랭이 10 농가에서 수집하여 수분활성도와 당도, 위생지표세균 (일반세균수, 대장균군, E. coli)과 병원성미생물(Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus), aflatoxin을 조사하였다. 감말랭이, 건시, 반건시의 수분활 성도는 각각 0.81, 0.86, 0.91 였으며 당도는 감말랭이 54.9%, 건시 49.5%, 반건시 40.5%로 당도와 수분활성도는 반비례 하였다. 곶감의 일반세균수는 감말랭이 3.93 ± 0.96 log CFU/g, 건시 2.12 ± 0.93 log CFU/g, 반건시 1.50 ± 1.08 log CFU/g로 감말랭이의 오염도가 가장 높았다. 또한 S. aureus는 감말랭이 40.0%, 건시 29.5%, 반건시 23.5%의 시료에서 검출되었고, E. coli는 건시 6.6%, 반건시 2.9% 의 시료에서 검출되었다. B. cereus는 건시 39.3%, 반건시 20.6%의 시료에서 검출되었고 검출 수준은 0.7~2.3 log CFU/ g이었다. Aflatoxin의 경우는 ELISA법으로 분석했을 때 감 말랭이, 건시, 반건시에서 각각 30%, 4.9%, 2.9%가 검출 되었고 검출농도는 1.0~1.1 ppb 이었다. ELISA법에서 양 성시료를 대상으로 UPLC로 재분석한 결과 aflatoxin은 검 출되지 않았다. 본 연구의 결과로 미루어 볼 때 안전한 곶 감 생산을 위한 위생관리기술의 개발과 곶감생산 농가의 지속적인 위생교육이 필요하다고 사료된다.

아플라톡신은 옥수수 같은 작물에 Aspergillus flavus와 A. parasiticus 같은 곰팡이에서 생성되는 독소로서 발암 등 식품 안전성에 심각한 문제를 일으키는 물질이다. 이런 곰팡이의 감 염 환경에 적게 노출된 우리나라와 달리, 아프리카 동부에 위 치하고 있는 우간다의 경우, 고온 다습한 기후에 의해 곰팡이 오염이 심각하며, 이에 따른 아플라톡신의 생성 또한 많아, 대 표적인 식량 수출국으로서 심각한 피해를 보고 있다. 이를 저 감하기 위해, 곰팡이 저항성 품종 육성, 수확 후 관리 기술 개발 등의 신기술이 필요한 실정이다. 본 연구에서는 곰팡이 저항성 신품종 옥수수 육종의 일환으로 우간다 현지 재배 옥수수에 피해를 주는 곰팡이 균주 조사 및 아플라톡신 오염 현 황을 조사하고자 하였다. 우간다 각지의 옥수수 생산 농가와 곡물 상점을 대상으로 55종의 시료를 확보하고, 이들에 대한 아플라톡신 오염을 조사하였다. 그 결과 총 시료 가운데 약 11% 이상의 시료에서 아플라톡신의 오염이 확인되었고, 이들 의 함량 또한 12.7 - 123.5 μg/Kg으로 우간다 현지의 규제 기 준을 초과하는 시료의 경우도 9% 이상 발견되었다. 이 수치 는 곰팡이 감염에 대한 대책 수립의 필요성을 뒷받침하고, 아 플라톡신 등의 곰팡이 독소의 생성을 억제 할 신품종 및 기술 개발이 필요함을 의미한다.

Mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) are widespread contaminants of food and feedstuffs. It is very likely, that humans and animals are always exposed to mixtures of mycotoxins rather than to individual compounds. Therefore, risk assessments should consider mixture toxicity data. In the present study the combination of AFB1, OTA and ZEA was tested for genotoxicity in rat bone marrow and blood leukocytes after 15, 30 and 60 days treatment. The level of DNA damage was determined by the comet assay. The tail intensity and Olive tail moment in leukocytes and bone marrow cells were significantly higher than in controls. At the same time, the level of DNA damage in bone marrow cells was higher than in leukocytes. The data suggests that prolonged exposure to mycotoxins combination through food consumption can induce DNA damage contributing to the harmful effects in vivo.

2012년 2월부터 11월까지 인천 지역에서 유통된 건고추 및 고춧가루 193건을 대상으로 아플라톡신 B1과 오크라톡 신 A의 오염도를 조사하였다. Immunoaffinity column 및 HPLC를 이용한 시험법은 모두 80% 이상의 회수율을 보였고, 아플라톡신 B1 및 오크라톡신 A의 검출한계는 각각 0.13 μg/kg, 0.30 μg/kg였다. 오염도 조사를 한 결과 아플라 톡신 B1은 17.1%의 검출율을 보였고 오크라톡신 A는 20.7% 의 검출율을 보였으며, 아플라톡신 B1의 검출농도는 0.14~ 9.67 μg/kg였고, 오크라톡신 A의 검출 농도는 0.31~3.31 μg/ kg였다. 이는 우리나라 식품공전 상의 기준인 10 μg/kg(아 플라톡신 B1), 7 μg/kg(오크라톡신 A)보다는 낮은 수치로 비교적 안전한 수준이었다.

The aim of this study is to develop the one dot rapid detection kit that meets the limit of detection (LOD) of KFDA (10 μg/kg) for Aflatoxin B1 (AFB1). The fabrication process of the one dot rapid detection kit, such as gold nanoparticle and AFB1-BSA conjugation, and AFB1-polyclonal antibody concentration was optimized. Fabricated rapid detection kits were tested with the standard AFB1 solutions and the solutions extracted from contaminated maize samples. The LOD in the standard AFB1 solution and extracted AFB1 solution was 5 μg/kg for both. This result is lower than the LOD of KFDA in food (10 μg/kg) and satisfies the LOD of EU (2-12 μg/kg) and Japan (10 μg/kg) in food.

아플라톡신 및 오크라톡신 A의 동시분석법을 확립하여보다 신속하고 경제적인 분석을 통해 아플라톡신과 오크라톡신 A를 효율적으로 관리하고자 하였다. 시료의 전처리는 80% methanol로 추출하고 면역친화성 컬럼을 이용하여 정제한 다음 trifluoroacetic acid를 이용하여 유도체화 한 후 HPLC-FLD로 분석하였다. 컬럼은 Capcell pak-C18(4.6 mm × 150 mm, 5 μm)을 사용하였으며, 이동상은water : acetonitrile : methanol (72 : 14 : 14) 용액과 acetonitrile: 0.1% phosphoric acid (50 : 50) 용액을 구배용매조성(gradient mode)으로 분석하였다. 형광검출기의 파장은 파장프로그램을 사용하여, 여기파장 360 nm, 검출파장 450 nm과 여기파장 330 nm, 검출파장 460 nm으로 분석하였다. 아플라톡신 B1, B2, G1, G2와 오크라톡신 A의 직선성을 검토한 결과 상관계수(R2) 0.9997~0.9998의 범위였고, 검출한계와 정량한계는 각각 0.05~0.18, 0.16~0.60 μg/kg이었다. 회수율은 아플라톡신 B1, G1, B2, G2, 오크라톡신 A 각각78.4~101.5%, 73.3~102.1%, 81.7~106.7%, 67.0~104.6%,78.7~120.8%이었다. 확립된 동시분석법을 통한 국내 유통식품을 대상으로 모니터링을 실시한 결과 총 151건 중 13건에서 아플라톡신및 오크라톡신 A가 검출되었으며 검출 범위는 아플라톡신B1은 0.32~1.80 μg/kg, 오크라톡신 A는 0.97 μg/kg이었다. 이들 결과로부터 우리나라에서 유통 중인 조사한 식품 151건의 아플라톡신과 오크라톡신 A의 검출량은 모두 기준치 이하로 적합한 것으로 조사되었으며 안전한 수준임을알 수 있었다.

Competitive ELISA methods were used to measure the level of aflatoxin M1 (AFM1) from urine in 1008 Korean adults. Subjects were selected by random sampling in all areas of Korea, except Cheju-do. The recovery rate of AFM1 using this method was 105% (73-124%). The geometric mean of urinary AFM1 in all subjects was 3.43 pg/mL (3.67 ng/g creatinine). The level of AFM1 in males was statistically higher, compared with female subjects. However, the levels of AFM1 did not differ according to age. Subjects in Chungbuk-do showed the highest urinary AFM1 concentration, whereas subjects in Kyeongnam-do showed the lowest concentration. Assuming an excretion rate of 5%, this AFM1 excretion corresponds to approximately 0.1 microgram/day in Korean adults.

Aflatoxins produced by Aspergillus spp. are recognized as a major concern in animal and human health. In pigs, ingestion of aflatoxin-contaminated feeds causes immunosuppression, hepatotoxicosis and poor feed efficiency. In this study, we screened the contaminated level of aflatoxins in 449 pig feeds from 12 swine farms in Korea. For rapid and efficient screening of aflatoxins in pig feeds we evaluated the feasibility of three commercial ELISA kits for the screening of aflatoxins in pig feeds. Twenty-nine pig feed samples were examined for total aflatoxins using three ELISA kits, simultaneously. From three repetitions of each assay, the average intra-assay precisions and the average inter-assay precisions expressed as coefficient of variation (CV, %) for VeratoxⓇ Quantitative Aflatoxin test were 6.90 and 12.29, respectively. A statistical comparison of the results between HPLC and ELISAs showed that the correlation coefficient values for VeratoxⓇ was 0.96. The results demonstrated that we can apply the VeratoxⓇ Quantitative Aflatoxin test for the detection of aflatoxins in pig feed from the field. The screening of field samples with this ELISA kit showed that 11 out of 265 pig feeds for growers and weaners were contaminated with total aflatoxin levels exceeding 10 ppb, maximum tolerable limits for their compound feeds and the aflatoxins levels of remaining 184 pig feeds for other age groups of pigs were confirmed as below 20 ppb. The results from the screening indicated overall low levels of aflatoxins contamination in pig feeds.

We used fluorescence detector to analyse total aflatoxins (G1, G2, B1, B2) with TFA (Trifluoroacetic acid) derivation method and PHRED (Photochemical reactor enhanced detection) method. PHRED method was superior in reproduction and convenience, but TFA derivation method was superior in selectivity and sensitivity. The recovery rate of aflatoxin B1, B2, G1 were more than 80%, and G2 was more than 70%. The detection limit of B1,B2, G1 and G2 were respectively 0.05, 0.05, 0.2 and 0.1 μg/kg. Confirmed method was used to analyse total aflatoxins in total 316 items as 9 kinds 137 dried fruits and 10 kinds 179 spices. By the result, Aflatoxins were detected in 27dried fruits (19.7%) and in 87 spices (48.6%).

This study was conducted to determine the effects of vitamin C on the activity of liver function enzymes and electromicrographic changes in white rats treated with aflatoxin B1(AFB1) or X-ray and AFB1. Six week-old male Sprague-Dawley rats were randomly divided into five groups: a control group, AFB1 treated group, AFB1 treated group with vitamin C, X-ray and AFB1 co-treated group, X-ray and AFB1 co-treated group with vitamin C. On the first day of the experiment, only one dose of X-rays was exposed to the entire liver at 1, 500 cGy. Next, vitamin C was injected at 10㎎/㎏body weight by intraperitoneal injection, followed 1 hr later by the administration of 0.4㎎/㎏of AFB1 by intraperitoneal injection. These treatments were then administered every three days over a period of 15 days. On the 16th day of treatments, the animals were sacrificed. Analysis of the activity of the liver function enzymes, GOT, ALK phatase and LDH, in the sera of rats revealed that they were somewhat increased by AFB1 treatment, X-ray and AFB1 co-treatment when compared to the control group. Furthermore, the activity of these enzymes decreased in response to administration of vitamin C. Especially, the levels of GOT were remarkably decreased in the AFB1 treated group treated with vitamin C when compared to the group treated with AFB1 alone(p<0.001). Electromicrographic analysis revealed cloudy swelling, necrosis, vesicular degeneration and fat accumulation of hepatocytes in response to treatment with AFB1 or co-treatment with X-ray and AFB1. However, the destruction of hepatic cells was considerably lower in the vitamin C-treated group. These results indicate that vitamin C had ameliorating effects on the hepatic cell damage.

Aflatoxin (AF) and Ochratoxin A (OTA) are carcinogenic and possible carcinogenic mycotoxins respectively produced by Aspergillus spp or Penicillium spp. The study for contamination survey and proposal for reduction of mycotoxin in red pepper were carried out. 192 samples were collected at such various stages and markets as pre/post-harvest stages, internet shopping mall /super-market and small stakeholder mill/geographically indicated company. As only 2 samples were positive for aflatoxin, so contamination rate was 1.04%. In the meanwhile, contamination rate for ochratoxin A was 21.88% and a various amount of OTA was detected in 42 positive samples. 6 samples were found to be contaminated at higher level than 5 μgkg−1 for ochratoxin A, which was established recently as a maximum permissible limit in korea. There was no difference in degree of contamination with regard to cultivation type because any mycotoxin was not found at all in both organically and conventionally grown red pepper. But, there was statistically significant difference in the process of manufacturing. Finished products were OTA-contaminated at a level of 2.32 ± 6.54 μgkg−1(mean ± SD), even though OTA was not detected in deep frozen red peppers right after long term storage. And contamination for OTA was a level of 0.33 ± 0.91 μgkg−1(mean ± SD) in red paprika powder after uv sterilization, while the contamination for OTA was 2.78 ± 4.49 μgkg−1(mean ± SD) in non-uv sterilized powder. In addition, our investigation shows that higher OTA contamination occurred in some of famous brand products sold in super-market and domestic products than products collected through on-line shopping or from small stakeholder mills and imported products respectively, however, difference was not statistically significant.