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 performed to develop and optimize a rapid strip kit for the dual-detection of Aflatoxin B1 (AF) and Ochratoxin A (OTA). The strip kit is composed of sample pad, conjugation pad, nitrocellulose membrane, and absorbent pad. Manually spotted mycotoxin-BSA conjugates and anti-mouse IgG on nitrocellulose membrane were used as the test line and the control line, respectively. Conditions for rapid and easy detection of mycotoxins, conjugation between toxin-MAbs and nano Au particles, pretreatment method of pads, and composition of standards solutions were optimized. Feasibility of the strip kit to detect AF and OTA were evaluated with standard samples. Test results were acquired in 10 min and the detection limits were 100 ppm for AF and OTA, respectively.

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.

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.

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.

This study is an endeavor to evaluate the risk assessment of hazardous(aflatoxin B1) in medicines from oriental medical prescription which are circulated much recently. For that, twelve globular and granule types, seven liquid types of herbal medicine were bought to compare and analyze the content of aflatoxin B1, which are harmful to human body. Woo Hwang Cheong Sim Hwan of Aflatoxin B1 concentration lower than the standard accepted by all the products have been detected, B company(tradition) is the concentration of 1.24 ㎍/kg, C company 1.04 ㎍/kg, A company(tradition) and B company did not detect. And the general pill of aflatoxin B1 concentration lower than the standard accepted by all the products have been detected, S-1 is the concentration of 1.8 ㎍/kg, S-2 of 1.04 ㎍/kg, S-3 of 0.88 ㎍/kg, S-4 of 9.32 ㎍/kg, S-6 of 7.8 ㎍/kg, S-5 did not detect. All the products eundan allowed in the concentration of aflatoxin B1 levels were lower than detection, D company of 0.96 ㎍/kg, E company concentration was not detected. The liquid product of aflatoxin B1 concentration was found liwer than the standard accepted by all the product, L-3 concentration of 0.8 ㎍/kg, K-4 was detected in the 1.16 ㎍/kg, L-1 and L-2 is not detected, L-5 concentration of 15 ㎍/kg, L-7 is detected as 1.08 ㎍/kg and, L-6 was not detected.

동치미에서 분리한 유산균인 Leu. mesenteroides subsp.cremoris DLAB19 균주의 aflatoxin Bl에 대한 항 돌연변이원성 물질 생산을 위한 최적 조건을 조사한 결과, 탄소원으로 glucose를 첨가시 가장 높은 항 돌연변이 효과를 나타내었으며, 질소원으로서는 yeast extract 첨가시 항 돌연변이 효과가 우수하였다. 탄소원으로 glucose의 농도을 2 첨가시aflatoxin Bl에 대한 항 돌연변이 효과가

본 연구에서 8종의 생약 추출물이 A. parasiticus의 배양시 aflatoxin B1 생성에 미치는 효과를 조사하였다. 배지의 pH는 배양 3일 후에 모든 생약 추출물이 pH 4 이하를 나타냈으며 구기자, 오매, 계피, 두충은 배양 6일에 다시 pH 4 이상으로 상승하였고 이중 대추가 배양 기간 중 가장 낮은 pH를 나타냈다. 균체 생성량은 모든 실험군이 대조군보다 높았으며 갈근, 두충, 오미자, 대추, 오매,구기자, 목과의 순으로 나타났다. 이중 갈근이 최대 생성량을 나타냈으며 목과가 가장 낮은 생성량을 나타냈다. Aflatoxin B1은 갈근과 대추 추출물을 제외한 모든 실험군에서 생성이 억제되었다. 특히 계피, 오매, 두충 구기자, 오미자 추출물 에서 aflatoxin B1 생성이 현저히 저하되었으며 계피가 가장 큰 억제 효과를 나타냈다. 균체량이 많이 생성되면 aflatoxin B1 생성이 적어지고 균체량이 적게 생성되면 aflatoxin B1의 생성이 많아졌다. Aspergillus parasiticus에 의해 aflatoxin B1을 가장 적게 생성하는 계피와 가장 많이 생성하는 갈근 추출물의 총단백질 생성량은 계피는 3일째 (34.5%), 갈근 ext ract는 4일째 (36.4%) 총단백질의 함량이 최대가 되어 균체 내의 총단백질의 함량은 계피와 갈근 추출물을 첨가한 시험군이 게조군의 총단백질 함량 (32.7%)보다 약간 많았으며 aflatoxin B1 생성 및 축적이 최대가 되는 시기는 총단백질량이 최대가 되는 시기보다 대체로 1일 정도 늦게 나타났다.