• Food Science and Preservation
• /
• v.18 no.3
• /
• pp.374-382
• /
• 2011

Aflatoxin ($AFB_1$) is a potent hepatotoxic and hepatocarcinogenic mycotoxin in humans. It is also well-known to be accumulated in animal tissues via various metabolic pathways. This study was conducted to determine the effects of vitamin C on the residual $AFB_1$ in rat sera that were treated with radiation and $AFB_1$. Six week-old male Sprague-Dawley rats were randomly divided into five groups: a control group, $AFB_1$-treated group, the group treated with $AFB_1$ and vitamin C, the group treated with X-ray and AFB1, and the group treated with X-ray and $AFB_1$ 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 mg/kg body weight via intraperitoneal injection, followed 1 hr later by the administration of 0.4 mg/kg of $AFB_1$ via 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. The contents of $AFB_1$ in rat sera were determined via indirect competitive ELISA and HPLC method. In the quantitative analysis of $AFB_1$ in rat sera via ELISA, $5.17{\pm}0.34$ ng/mL of $AFB_1$ was detected in the $AFB_1$-treated groups, but the amount more significantly decreased to $3.23{\pm}0.76$ ng/mL in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. The $AFB_1$ contents of the rat sera of the groups treated with X-ray and $AFB_1$ did not significantly decreased with the administration of vitamin C. The $AFB_1$ content of the rat sera that was analyzed via HPLC showed a tendency similar to that of the content that was analyzed via ELISA. With regard to these data, vitamin C was very effective in reducing $AFB_1$ residue in rat sera.

• Journal of Life Science
• /
• v.23 no.2
• /
• pp.306-310
• /
• 2013

We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

• Journal of Food Hygiene and Safety
• /
• v.3 no.2
• /
• pp.89-97
• /
• 1988

The possible effeets of some oriental herbs, whleh have been used to treat eaneerllke dllleue In Korea, on growth and afIatonn production by Aspergillus parasiticus R-716 were investigated. Zedoaria rhizoma, Curcuma longa, Cyperus rotundus, Angelica gigas, Paeonia albiflora, Paeonia mountan, Atractylis ovata, and Pulsatilla koreana were extracted with chloroform. Among them the extract of paeonia mountan was remarkably effective on the growth inhibition, and Curcuma longa, Zedoaria rhizoma, Cyperus rotundus, Paeonia albiflora, Atractylis ovata also inhibited the growth. The ertraet of Atractylis ovata and Curcuma longa, also Inhibited the afIatoxin production but the others showed no effect at all or sometimes stimulated effect. With the addition of O.2ml extract of Atractylis ovata in 30ml SLS medium, the growth was delayed for about 2 days, and after 9 days, mycellum weight was 0.953g, and total afatoxin was reduced 5O%($792\mu\textrm{g}$)of that produced in the control($1547\mu\textrm{g}$). Aflatoxin per mycelium weight was decreased 32%($992\mu\textrm{g}$) of that produced in the control($1467\mu\textrm{g}$), but NADPH oxidase was higher as compared to the control. The extract of Atractylis ovata appeared to have a inhibitory effect on the growth and the aflatoxin production of Aspergillus parasiticus R-716.

• Journal of Food Hygiene and Safety
• /
• v.16 no.4
• /
• pp.274-279
• /
• 2001

Generally, non-aflatoxigenic fungi, such as Aspergillus oryzae, and Aspergillus are main microflora in Korean traditional fermented foods including Meju and soybean paste, but sometimes, Aspergillus flavus and Aspergillus parasiticus can be contaminated and accumulated aflatoxins during fermentation and storage. So the screening of aflatoxigenic strains in fermented traditional food is very important to improve the sanitary quality of those foods. In this work, we screened aflatoxin producing fungi from commercial Meju and soybean paste in Western Gyeongnam by immunoassay. Samples were randomly purchased from market of the commercial Meju(10 EA) and soybean paste(20 EA) in nine areas of Western Gyeongnam. Of the samples collected,24 strains and 22 strains of Aspergillus sp. were isolated from Meju and soybean paste, respectively. The isolated strains were cultured on SLS media at $25^{\circ}C$ for 15 days. The cultured broth were extracted with ethyl acetate and were analysed to determine aflatoxin B$_1$(AFB$_1$) by direct competitive ELISA(DC-ELISA). Six strains(25%) isolated from Meju, and 2 strains(9%) isolated from saybean paste, were confined as aflatoxin producing strains. The average range of aflatoxin productivity of isolates from Meju was 54.6 $\pm$ 38.7 ng/ml and that from soybean paste was 11.1 $\pm$ 8.6 ng/ml, respectively. Among them, isolated strain No. M-5-4 produced a high level of AFBl and showed 98.26 ng/ml of AFB$_1$. Every isolates were also re-confined their AFB$_1$productivity by thin layer chromatography(TLC). The TLC results also showed same trend as DC-ELISA results. As the above results, the screening of hazard mycotoxigenic fungi from traditional fermented foods should be necessary for the safety and the application of HACCP system in the food manufactory in Korea.

• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.7
• /
• pp.1011-1018
• /
• 2011

Aflatoxin-contaminated feed cause mortality, suppression of the immune system, reduced growth rates and losses in feed efficiency. This research study was planned to investigate the immunomodulatory and growth promoting effect of milk thistle as feed additive against aflatoxin $B_1$ in broiler chicks at NWFP Agricultural University Peshawar, Pakistan. Two hundred and forty (240) day old broilers chicks were randomly assigned into four major groups AfF, aflatoxin free feed; Aflatoxin $B_1$ was present in the feed at the levels of 80-520 ${\mu}g/kg$ of the feed in the remaining three groups. Aflatoxin contaminated feed was provided for 5 weeks. Group AfB was supplemented with toxin binder "Mycoad" at 3 g/kg of feed and group AfT was supplemented with milk thistle at10 g/kg of feed. Each group was further sub divided into two sub-groups, vaccinated against ND (Newcastle disease), IB (Infectious bronchitis) and IBD (Infectious bursal diseases) according to recommended schedule of vaccination or non vaccinated. Each sub group carried three replicates with 10 chicks per replicate. Chicks were reared in pens in an open sided house. Supplementary heat was provided to all the chicks during brooding period. Mean body weight gain and dressing percentage were significantly (p<0.05) higher in group AfF, followed by AfT, AfB and Af. Weight gain and dressing percentage was the same in group AfB and AfT, while it was significantly lower in group Af. Feed intake, breast, thigh and leg weight were found significantly (p<0.05) higher in group AfF, followed by AfB, AfT and Af. Significantly lower (better) FCR value was recorded in group AfT. Water intake was significantly (p<0.05) higher in group AfT and AfF as compared to other groups. Mortality was significantly (p<0.05) higher in group Af. Mean bursa and thymus weights were found significantly (p<0.05) higher in group AfF, AfB and AfT followed by Af, while higher spleen weight was recorded in group AfT. Mean antibody titer against ND, IB and IBD was significantly (p<0.05) higher in group AfT, as compared to other groups. It is concluded that milk thistle at 10 g/kg of feed could effectively be utilized as immunostimulant and growth promotant in the presence of immunosuppressant aflatoxin $B_1$ in the feed.

• The Plant Pathology Journal
• /
• v.18 no.4
• /
• pp.204-209
• /
• 2002

Andhra Pradesh is one of the major groundnut growing states in India. A total of 182 groundnut samples collected at harvesting from farmers fields in five districts, namely; Anantapur, Chittoor, Cuddapah, Kurnool, and Mahaboobnagar, during 1999 and 2000 rainy seasons were evaluated for the presence of mycotoxins (both aflatoxins and zearalenone) and toxigenic fungi. In samples collected from each district, average seed infection by Aspergillus flavus and Fusarium spp. was 11.9-18.3% and 5.6-12.8% in 1999, and 9.5-14.1% and 9.4-11.9% in 2000, respectively. Among the samples collected, 20.3% and 16.5% were contaminated with aflatoxin in 1999 and 2000, respectively, and in 11.4% and 8.7% of the seed samples collected in two seasons, the aflatoxin content was >30 $\mu\textrm{g}$/kg. An alarming aflatoxin content of 851.9$\mu\textrm{g}$/kg was found in samples collected from Anantapur district during the rainy season in 1999. Zearalenone was not detected in any of the samples collected in 1999, while 2 out of 103 samples collected in 2000 were contaminated with 35.1 and 129.4$\mu\textrm{g}$/kg. Under in vitro cultural conditions, 35.8% of the 173 A. flavus isolates collected from the groundnut samples produced aflatoxins at concentrations of 94.3-1598.6 ng/$\textrm{m}{\ell}$ and 3% of the 266 Fusarium spp. isolates produced 98.1-847.3 $\mu\textrm{g}$/g of zearalenone. The results emphasize the need for a more systematic and regular monitoring of pre-harvest aflatoxin contamination.

• Journal of Food Hygiene and Safety
• /
• v.18 no.3
• /
• pp.95-100
• /
• 2003

Competitive direct enzyme-linked immunosorbent assay(cdELISA) of aflatoxin $B_1$ ($AFB_1$) in deonjang(Korean-style soybean paste) and kochujang(fermented hot peppersoybean paste) and the level of $AFB_1$ in modern or traditional style deonjang and gochujang, produced in Korea, was surveyed by cdELISA. From the standard curve of the cdELISA, the detection limit of $AFB_1$ was 0.2 ng/m/. The average recovery of $AFB_1$ was 71.5% in the range of 1~100 ng/g after spiking $AFB_1$ into deonjang and it means that it could be possible to detect the $AFB_1$ in these range by the cdELISA in deonjang. Among the 30 kochujangs tested, no $AFB_1$ was detected in kochujangs. Among the 30 deonjangs, $AFB_1$ was detected in 6 ones in the range of 1.0~6.0 ng/g. The occurrence of $AFB_1$ in deonjang and kochujang tested in this study was less than the Korea Standard and Specification of aflatoxin in foods (10 ppb).

• Journal of Food Hygiene and Safety
• /
• v.16 no.3
• /
• pp.200-205
• /
• 2001

In this study, occurrence of aflatoxin M$_1$(AEM$_1$) in domestic milk and milk products was determined. The level of AFM$_1$ in market milk (0.047 ppb) was lower than that in raw milk (0.083 pub) but this looks like that is due to dilution in collecting process rather than the effect of sterilization. In the case of nonfat dry milk, level of AFM$_1$appeared high by 0.24 ppb but it is thought to be not different from market milk actually because nonfat dry milk is diluted at intake. In the case of ice cream, finished products were contaminated with AFM$_1$of 0.020 ppd and also have the possibility of the contamination of AFB$_1$due to secondary raw material such as nuts and almond. On the basis of the results of this study and previous studies, Monte-Carlo simulation is conducted to estimate the contamination level of AFM$_1$in domestic market milk. To consider uncertainty and variability fitting procedure was passed through. And we used beta distribution to estimate the prevalence and triangular distribution to estimate the concentration level of AFM$_1$in milk. As a result, the 5%, 50% and 95% points of the distribution of the probability of AFM$_1$contamination level in milk is 0.0214, 0.0946 and 0.1888 ppb, respectively. Also we estimate that AFM$_1$in almost milk was low more than 0.5 ppb that is American acceptable level but 80.4% exceeded far 0.05 ppb that is European standard.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.10
• /
• pp.1629-1637
• /
• 2007

An immunochromatography (ICG) strip test using a nanocolloidal gold-antibody probe was developed and optimized for the rapid detection of aflatoxin B1 (AFB1). A monoclonal antibody specific to AFB1 was produced from the cloned hybridoma cell (AF78), coupled with nanocolloidal gold, and distributed on the conjugate pad of the ICG strip test. The visual detection limit of the ICG strip test was 0.5 ng/ml, and this method showed a cross-reaction to aflatoxin B2, G1, and G2. In total, 172 grain and feed samples were collected and analyzed by both the ICG strip test and HPLC. The results of the ICG strip test showed a good agreement with those obtained by HPLC. These results indicated that the ICG strip test has a potential use as a rapid and cost-effective screening tool for the determination of AFB1 in real samples and could be applied to the preliminary screening of mycotoxin in food and agricultural products, generating results within 15 min without complicated steps.

• Toxicological Research
• /
• v.32 no.1
• /
• pp.81-87
• /
• 2016

Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was $65^{\circ}C$. The optimized template and primer concentration were $1.5{\mu}L\;(50ng/{\mu}L)$ and $3{\mu}L\;(10{\mu}M/{\mu}L)$ respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at $88.0^{\circ}C$, $87.5^{\circ}C$, $83.5^{\circ}C$, and $89.5^{\circ}C$ respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.