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.

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.

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.

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.

된장과 고추장에 대해 cdELISA에 AFB_(1)의한 의 분석방법을 확립하고 국내에서 생산되는 재래식 및 개량식 된장과 고추장을 수거하여 AFB_(1)의 오염도를 조사하였다. 표준곡선을 작성하였을 때, 이의 검출 한계는 0.2ng/㎖(ppb)이었다. 된장의 Spike test 후 회수율은 1~100 ng/g 범위에서 평균 71.5%로 나타나 이 범위에서 cdELISA에 의한 AFB_(1)의 측정이 가능함을 보여주었다. 이로부터 된장 및 고추장에서 cdELISA를 통한 AFB_(1)의 분석 방법을 확립하였다. 된장 및 고추장 시료의 AFB_(1)의 오염도는 총 30종의 고추장 시료에서 AFB_(1)이 검출되지 않았고 총 30종의 된장 시료 중 6종에서 1.0~6.0 ng/g 수준으로 검출되었다. 이는 우리 나라의 허용기준인 10 ng/g(ppb) 이하였다.

Lipid peroxidation is one of the main manifestations of oxidative damage and has been found play an important role in the toxicity and carcinogenesis of many carcinogens. This study was carried out to investigate the effects of aflatoxin B₁ co-administrated with antioxidant vitamins on lipid contents and fatty acids components of liver in mice. For this work, vitamin C and vitamin E, the major antioxidants, were administrated with 10 ㎎/㎏ and 63.8 ㎎/㎏ respectively, through intraperitoneal(i.p) injection to male ICR mice, and 0.4 ㎎/㎏ of the AFB₁ injected by i.p. lhr later. The results were as follows: two fold amounts of free cholesterol, triglyceride, and total cholesterol in serum and liver of mice treated with only AFB₁ were observed, when compared to those of mice co-administrated with antioxidant vitamins. However, the levels of phospholipids in serum and liver of mice treated with only AFB₁ were decreased. Concerning to fatty acids composition of liver from AFB₁-treated mice, P/S ratio was shown more low level in cholesteryl ester, triglyceride, total cholesterol and phospholipid than those of mice co-administrated with antioxidant vitamins. In these data which provide with a reliable evidence on their antioxidantal effects to aflatoxicosis.

쌀에 대하여 aflatoxin 생성을 위한 기질로서의 능력을 알아보기 위하여 현미(청청벼) 시료에 Aspergillus parasiticus를 접종하고 조건을 달리하여 저장하면서 aflatoxin B₁의 생성을 관찰하였다. 시료중의 aflatoxin B₁의 분석은 ELISA를 이용하여 수행하였다. 현미 시료에서 aflatoxin B_1 생성에 가장 좋은 온도는 28℃였으며, 시료의 수분함량을 15.8%로 증가시킨 경우 aflatoxin B₁의 생성이 유의하게 증가하였고 (p<0.05), 고압증기멸균시킨 시료는 aflatoxin B_a 생성에 보다 효과적인 기질이 되었다. 실온에서 3개월 동안 저장한 현미에서는 15일 동안 저장한 경우에 비하여 aflatoxin B₁ 생성이 유의한 증가를 보였다(p<0.05). 따라서 저장온도 및 수분함량이 쌀에서도 aflatoxin B₁ 생성에 영향을 미치는 바를 나타내었으며, 또 시료의 상태 및 저장기간도 쌀에서 aflatoxin 생성 위험요인으로 작용할 수 있음이 제시되었다. 이러한 결과로부터 쌀이 aflatoxin 생성에 좋은 기질이 될 수 있는 것으로 평가된다.

Drosophila의 actin 5C 유전자 promoter에 쥐의 DNA polymerase cDNA를 도입시킨 형질전환 초파리가 고감수성 환경성 변이원 검출계로 사용할 수 있는지를 조사하였다. 체세포 염색체 재조환과 체세포 염색체 돌연변이의 검출을 위해서는 geterozygous(mwh/+) 계통을 사용하였다. 염색체상의 결실이나 비분리 등에 의한 small mwh spot의 자연 발생적 빈도는 non-transgenic w 계통과 transgenic p[pol ]-130 계통에서 각각 0.351 및 0.606 정도였다. 체세포 염색체 재조환에 의한 large mwh spot의 자연 발생적 빈도의 경우는 transgenic p[pol ]-130 계통(0.063)이 non-transgenic w 계통(0.021)에 비해 약 3배 정도 높게 나타났다. IQ, Glu-P-1 및 TEXAFB1/TEX 등의 돌연변이원의 처리에 의한 경우, 두 종류의 mutant clone의 발생 빈도는 쥐의 DNA polymerase 가 도입된 transgenic p[pol ]-130 계통이 non-transgenic w 계통에 비하여 모두 약 2-3배 정도 높게 나타났다. 본 연구의 결과는 쥐의 DNA polymerase 가 최소한 체세포 염색체 돌연변이 유발이나 체세포 염색체 재조환의 생성 과정에 관여함을 의미하며, 형질전환 초파리 계통이 환경성 변이원 검출계로서 충분한 응용가능성이 있음을 보여 주었다.

The antimutagenic effects of 46 kinds of medicinal plants that have been used as traditional folk antitumor agents in Korea were studied by using Ames mutagenicity teat. Most of the methanolic extracts from the plants which were used in this experiment showed strong antimutagenic activity toward atlatoxin B₁(AFB₁) in Salmonella typhimurium TA100 and TA98. However, N-methyl-N'-vitro-N-nitrosoguanidine (MNNG) induced mutagenicity was not blocked by adding the methanolic extracts of the plants except persimmon leaves (Diospyros kaki Thunberg) and Elaeagnus umbellata.

Daily exposure of aflatoxin B₁ (AFB₁) was estimated in foods (rice, barley, soybean, peanut, soysauce, soybean paste) by ELISA (enzyme linked immunosorbent assay) using polyclonal antibody R_(101). Before ELISA, a simple extraction method was applied for the quantitation of AFB₁ in foods using chloroform which showed high recovery (70± 12%). AFB₁ levels in foods were 0.32 ng/ml (rice), 0.24 ng/ml (barley), 0.22 ng/ml (peanut), 0.30-0.78 ng/ml (soysauce), and 0.2 ng/ml (soybean paste). Based on food consumption, we estimated that Koreans were exposed to AFB₁ at the level of 1.86±0.46 ng/kg/day and liver cancer incidence attributed to AFB₁ exposure (assuming that AFB₁ as a single hepatocarcinogenic agent) might be calculated to be 13.1 per 100,000 population. Our data demonstrate that AFB₁ levels in foods were below the regulation of 10 ppb in foods and might not be the major risk factor for the high incidence of liver cancer in Korea.

본 연구는 흰쥐에게 AFB1을 투여하거나 방사선과 AFB1을 병합처리함으로 유발된 흰쥐의 간세포에서의 AFB1-DNA 부가체의 형성과 세포의 산화적 손상에 대한 vitamin C의 효과를 조사하기 위하여 수행되었다. X-ray 조사는 실험기간 내 단 1회로 실험사육기간 1일에 조사 하였고 X-ray 조사 후 vitamin C를 투여하였으며 vitamin C 투여 1시간 후 AFB1을 투여하였다. Vitamin C와 AFB1은 모두 복강투여로 실험 사육 첫 일부터 1회 시작하여 3일에 한번씩, 5회 반복 투였으며 실험동물 사육기간은 총 15일로 하였다. ELISA에 의한 흰쥐의 혈청 내 AFB1 잔여 농도는 AFB1 단독 투여군에서 5.17±0.34ng/mL이었으나 여기에 vitamin C 혼합 투여군에서는 3.23±0.76ng/ml가 검출되었다. 간세포의 AFB1-DNA adduct 농도는 AFB1 단독 투여군에서는 9.38±0.41ng/mL이었으며 2군에 vitamin C를 함께 투여한 3군에서는 5.28±0.32ng/ml로 나타나 2군에 비해 유의적으로(p＜0.001) 44% 감소한 양상을 나타내었다. 한편 X선 조사와 AFB1 병합처리한 4군에 비해 4군에 vitamin C를 투여한 5군에서 혈청 내 AFB1 함량과 간세포의 AFB1-DNA adduct 함량이 다소 감소하였으나 유의적인 차이는 없었다. 또한 면역조직화학적 관찰에서 AFB1 단독 투여군에서는 중심정맥과 혈관주변에서 AFB1 축적이 관찰되었는데 이러한 현상은 vitamin C를 혼합 투여함으로써 중심정맥과 혈관 주변의 갈색 침전이 현저하게 감소한 것으로 나타났다. 그러나 X선 조사와 AFB1 병합 처리한 군에서는 그 정도가 약했다.

Atlatoxin은 Aspergillus속 곰팡이로부터 생성되며 사람에게 있어서 간독성, 및 간암을 유발하는 잠재력을 가진 대표적 곰팡이 독소이며, 생체에 섭취된 은 여러 가지 대사경로를 거쳐 동물 조직 내에 축적되는 것으로 알려져 있다. 방사선은 수술, 항암약물요법과 더불어 임상 시 중요한 치료방법이나 정상세포에 방사선을 조사하였을 때 반응성이 높은 활성산소와 과산화라디칼()을 생성하여 세포막의 불포화 지방산을 지질 과산화물로 변성시켜 세포 산