Survey of aflatoxin contamination of dried figs grown in Turkey in 1986

A total of 284 dried fig samples, collected from fields during drying, and from warehouse and processing units in the Aegean region of Turkey in 1986, were examined for aflatoxin contamination. Aflatoxin B1, B2, and G1 were detected in 4, 2, and 2% of the samples, respectively, which were of the lower grade of figs taken from the drying stage. The average alfatoxin levels in positive samples were estimated to be 112.3 (B1), 50.6 (B2), and 61.4 ng/g (G1). The samples collected from storage (64 samples) and processing units (14 samples) contained no aflatoxins. The results of this survey show that aflatoxin contamination of Turkish dried figs in 1986 was highly correlated with the poorer grade of fig.     

Survey for co-occurrence of ochratoxin A and aflatoxin B1 in dried figs in Turkey by using a single laboratory-validated alkaline extraction method for ochratoxin A

A survey was carried out to determine the co-occurrence of ochratoxin A and aflatoxin B1 in dried figs from Turkey. Samples from two seasons of crops (2003 and 2004) intended for export to the European Union and the 2004 crop obtained from the domestic Turkish market were analyzed. Affinity column cleanup methods were employed for determining separately ochratoxin A and aflatoxin B1, but for ochratoxin A an alkaline extraction procedure was employed (in contrast to the conventionally employed acidic extraction), which gave consistently higher toxin recovery. In-house validation of the ochratoxin A method gave a limit of detection of 0.15 ng/g and a limit of quantification of 0.5 ng/g with a repeatability of 5.8% in the range 5 to 10 ng/g (with a mean recovery of 94% for spiked samples). Positive results for ochratoxin A were confirmed by liquid chromatography-mass spectrometry. For the 2003 export figs (58 samples), 7 samples contained only aflatoxin B1, 2 samples contained only ochratoxin A, and 2 samples contained both toxins (with maximum concentrations of 35.1 ng/g for aflatoxin B1 and 13.0 ng/g for ochratoxin A). Similarly for the 2004 export figs (41 samples), 16 samples contained only aflatoxin B1, 4 samples contained only ochratoxin A, and 2 samples contained both toxins (with maximum concentrations of 20.6 ng/g for aflatoxin B1 and 26.3 ng/g for ochratoxin A). Of 20 retail samples of dried figs from Turkey, only one sample contained ochratoxin A (2.0 ng/g) and none were contaminated with aflatoxin B1. This survey revealed a 14 to 15% incidence of occurrence of ochratoxin A for 2 years, which is higher than previously reported.     

The relationship between moisture content and equilibrium relative humidity of dried figs

A vacuum oven method is recommended for the determination of the moisture content of figs, by drying sliced quarters of whole figs at 70°C, at a pressure of less than 5 mm Hg for 24 hr. There is a greater variation between the moisture content of whole figs than between quarters of a single fig and the mean moisture content of a sample of figs can be determined with a standard error of 0.2–0.3%, by taking one quarter each from a number of whole figs. Data are presented for the moisture content/equilibrium relative humidity (e.r.h.) relationship of Turkish Lerida figs.     

THE AFLATOXIN CONTAMINATION OF FIG FRUITS IN AYDIN CITY (TURKEY)

The mold flora of 50 dried fig samples consumed in Turkey was examined and the aflatoxigenic ones were determined. Among 127 fungi isolated, 74 were Aspergillus, 24 were Trichoderma, 16 were Fusarium and 13 were Acremonium. Of the isolates, 17 were aflatoxigenic and four of them were capable to produce aflatoxin, three of which were characterized as A. flavus and one as A. parasiticus. Aflatoxin production of four strains was confirmed by high pressure liquid chromotography. The effect of UV irradiation on mold count and aflatoxin quantity was also tested. It was found that UV irradiation led to a decrease in the mold count and aflatoxin quantity.

PRACTICAL APPLICATIONS

Studies have shown that the concentration of aflatoxins may exceed the determined limits in dried figs. Its presence can be a potential threat to the health of consumers. Dried figs are one of the major agricultural export products of Turkey ( Senyuva et al. 2005 ). The effects of UV irradiation on mold flora of dried figs and aflatoxins have been examined. The Aspergillus flavus and parasiticus agar (AFPA) medium is used for detection of aflatoxigenic species, and coconut agar medium (CAM) is used to detect the aflatoxin-producing ability of aflatoxigenic strains. It was found that the reproduction of the molds in dried figs, consequently the aflatoxigenic mold strains, can be depressed by UV irradiation. It was found that increasing time of UV irradiation led to a decrease in the mold count in dried figs. In addition, a UV irradiation applied for 90 min, was found to decrease the aflatoxin quantity in dried figs in an amount of 25%. Because of inexpensiveness and easiness of the application it was concluded that the UV irradiation can be used as a practical application.     

Mycoflora and natural occurrence of aflatoxin, cyclopiazonic acid, fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B(1) (FB(1)) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range = 0.1-15.3 ng g(-1)), 74.7% of the samples had FB(1) (range = 0.05-3.65 mg kg(-1)), 10.0% of the samples had aflatoxin (range = 0.1-763.2 ng g(-1)) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range = 25-187 ng g(-1)). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.     

Aflatoxins, patulin and ergosterol contents of dried figs in Turkey.

Dried figs of three different categories, palatable, fluorescent, and cull, were investigated for their contents of aflatoxins (B(1), B(2), G(1) and G(2)), patulin, and ergosterol. Samples were obtained from four fig processing plants located in a major fig producing area in the Aegean Region in Turkey. Affinity column clean-up methods were employed for aflatoxins. All aflatoxins, patulin, and ergosterol were determined using high performance liquid chromatography. Palatable figs contaminated with trace amounts of aflatoxins, patulin, and ergosterol, so they posed no risk for the consumer when national and/or international regulatory limits were considered. Fluorescent figs were contaminated with high (117.9-471.9 ppb) aflatoxin levels and cull figs with high patulin (39.3-151.6 ppb) and ergosterol (4.5-18 ppm) levels. The total aflatoxins content was significantly correlated with the patulin content (r(2) = 0.813, p < 0.002) and the ergosterol content (r(2) = 0.920, p < 0.002) only in fluorescent figs. However there was no significant correlation between patulin and ergosterol contents of fluorescent figs. Furthermore, there were no significant correlations between the contents of any two of the three substances in cull figs. This is the first report on the presence of patulin and its co-occurrence with aflatoxin in dried figs.     

Aflatoxins in Turkish dried figs intended for export to the European Union

Dried figs for export from Turkey from crop years 2003 through 2006 were tested for aflatoxin B1 and total aflatoxins. For export to the European Union, consignments of 0.5 to 10 tonnes of dried figs were sampled according to European Commission regulations, and high-pressure liquid chromatography (HPLC) was used to determine concentrations of aflatoxins Bl, B2, G1, and G2. For each consignment of dried figs, a 30-kg sample (comprising 100 subsamples) was divided into three 10-kg subsamples, which were separately blended and analyzed with HPLC. This monitoring effort was conducted for figs from 2003, 2004, 2005, and up to June 2006, for a total of 10,396 30-kg samples (28,489 analyses). The incidence of contamination with aflatoxin B1 at higher than 2 ng/g was on average 0.6, 2.0, 4.0, and 2.4% for 2003, 2004, 2005, and up to June 2006, respectively, whereas contamination with total aflatoxins at higher than 4 ng/g was 2.6, 3.0, 5.1, and 2.7%. There was significant variability in contamination between replicate 1-kg samples, indicating small numbers of individual contaminated figs were probably responsible. There were also substantial differences in the relative proportions of aflatoxins B1, B2, G1, and G2 among samples, suggesting different contributing fungal sources.     

高特异性黄曲霉毒素B_1单克隆抗体的制备及特性研究

本研究将黄曲霉毒素B1转化为半缩醛B2a,在硼氢化钠（NaBH4）还原作用下与载体蛋白偶联制备完全抗原。将制备的完全抗原免疫Balb/c小鼠,经4次免疫后取其脾脏与小鼠骨髓瘤细胞Sp2/0细胞融合,采用半固体培养基筛选后鉴定,获得杂交瘤细胞株3A12,抗体的灵敏度可达6.1±0.025ng/mL,抗体与其它黄曲霉毒素B2、G1及G2的交叉反应率依次为7.8%、20.2%及0.6%,与黄曲霉毒素M1交叉反应率小于0.1%。本研究为研发花生等农产品黄曲霉毒素B1特异性免疫分析技术及产品奠定了重要基础。     

Aflatoxin and Cyclopiazonic Acid Production by Aspergillus flavus Isolated from Contaminated Maize

Seven truck-loads of maize were tested for mycotoxin contamination. Aflatoxin was identified in all 7 at concentrations from 3 ng/g-501 ng/g (aflatoxin B₁+ B₂). Cyclopiazonic acid was identified in 4 loads with concentrations from 25-250 ng/g. Deoxynivalenol was found in 4 of 5 loads tested, over a range of 46-676 ng/g. Ninteeen isolates of Aspergillus flavus from the samples were tested for ability to accumulate cyclopiazonic acid and aflatoxin in liquid culture. Fourteen produced cyclopiazonic acid (0.5-135 μg/mL), 12 produced aflatoxin (0.01-0.70 μg/mL, aflatoxin B₁+ B₂), and one aflatoxin-producing isolate did not produce cyclopiazonic acid.     

Effect of microwave heating during alkaline-cooking of aflatoxin contaminated maize

To evaluate the effectiveness of maize detoxification achieved with a modified tortilla-making process (MTMP), maize contaminated with aflatoxin B(1) (AFB(1)) and aflatoxin B(2) (AFB(2)) at levels of 22.46, 69.62, and 141.48 ng/g (AFB(1)+ AFB(2)) was processed into tortillas. Aflatoxin content was determined according to the 991.31 AOAC official method. Based on the results obtained with spiked samples (0.78 to 25 ng/g), the mean recovery was 92%, with a standard error of 1.2, and a coefficient variation value of 4.4%. The MTMP caused 68, 80, and an 84% decrease in aflatoxin content, respectively. Extract acidification (as occurs during digestion) prior to mycotoxin quantification caused some reformation of the aflatoxin structure in tortillas (up to 3%). According to these results, the MTMP seems to be safe for decontamination since a low percentage of the initial aflatoxin concentration can be reverted to the original fluorescent form upon acidification. PRACTICAL APPLICATION: The potential presence of aflatoxins in maize destined for human consumption is a serious problem to the Mexican food supply, as these toxic compounds may persist during the traditional alkaline-process for tortilla elaboration. Consequently, new detoxification procedures are needed that eliminate or at least minimize the aflatoxin risk, through lowering aflatoxin concentration in maize-based products. Under these considerations, the use of MTMP is recommended, since it has definite advantages including non-production of wastewater and reduced energy/time consumption.     

Aflatoxin contamination in food commodities in Bangladesh

During September 2009, we performed a rapid cross-sectional study to investigate the extent of aflatoxin contamination among common Bangladeshi foods. We collected eight common human food commodities (rice, lentils, wheat flour, dates, betelnut, red chili powder, ginger and groundnuts) and poultry feed samples from two large markets in each of three cities in Bangladesh. We quantified aflatoxin levels from pooled subsamples using fluorescence high-performance liquid chromatography. Aflatoxin levels were highest in dates and groundnuts (maximum 623 and 423?ng/g), respectively. Samples of betelnut (mean 30.6?ng/g), lentils (mean 21.2?ng/g) and red chili powder (>20?ng/g) also had elevated levels. The mean aflatoxin level among poultry feed samples was 73.0?ng/g. Aflatoxin levels were above the US maximum regulatory levels of 20?ng/g in five of eight commonly ingested human food commodities tested.     

Degradation of aflatoxin by lactoperoxidase

Summary Three concentrations of lactoperoxidase, 5, 50, and 500 units/ml of reaction mixture, degraded aflatoxin in the presence of 225 M NaCl and 50 M H₂0₂ at 28° C. Increasing the amount of lactoperoxidase from 50 to 500 units/ml of reaction mixture resulted in increasing the rate of degradation of aflatoxin B₁ from 3.6 to 5.1%/24 h. When comparable amounts of lactoperoxidase were present, aflatoxin G₁ was degraded approximately 1.5 times faster than was aflatoxin 131. At a given concentration of lactoperoxidase, aflatoxin degradation was independent of initial aflatoxin concentration. Derivatives that cochromatographed with aflatoxin B_2a and derivatives that were water soluble were the major degradation products of aflatoxin B₁. Similar derivatives, but in greater proportions, were noted as degradation products that resulted from activity of a blendure of mycelia ofAspergillus parasiticus.

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Abbau von aflatoxin durch lactoperoxidase

Zusammenfassung Aflatoxin wurde in Gegenwart von 225 m-NaCI und 50 m-H₂O₂ bei 28° C durch 5, 50 bzw. 500 Einheiten von Lactoperoxidase/ml Reaktionsgemisch abgebaut. Wenn die Konzentration der Lactoperoxidase von 50 bis auf 500 Einheiten/ml Reaktionsgemisch gesteigert wurde, dann stieg die Geschwindigkeit des Aflatoxinabbaues von 3,6 bis auf 5,1%/24 Std. Lactoperoxidase baute Aflatoxin G₁ 1,5 mal schneller ab als Aflatoxin B₁. Die Konzentration des Aflatoxins zu Beginn spielte keine Rolle beim Abbau durch, die Lactoperoxidase. Die Abbauprodukte von Aflatoxin B 1 waren chromatografisch dem Aflatoxin B_2a ähnlich oder waren wasserlöslich. Gleiche Abbauprodukte, aber in größeren Anteilen, erhielt man durch das Mycel vonAspergillus parasiticus.

     

Aflatoxin contamination of consumer milk caused by contaminated rice by‐products in compound cattle feed

BACKGROUND: Elevated levels of aflatoxin M1 were observed in routine checks of consumer milk in southern Sweden in early 2006. A trace‐back study revealed contaminated milk from several farms, and a total of 68 farms were banned from delivering milk to dairies for shorter or longer periods. The maximum level of aflatoxin M1 in a single sample from an individual farm was 257 ng kg^?1 fresh milk. RESULTS: Aflatoxin analyses of commercial compound feed revealed that the contamination originated from the ingredient rice feed meal, a by‐product from the preparation of Basmati rice for human consumption. Up to 56 µg kg^?1 of aflatoxin B1 was found in rice feed meal at one feed mill. CONCLUSION: The present example shows that an aflatoxin‐contaminated minor feed ingredient included at less than 10% (w/w) of compound cattle feed can significantly contaminate the milk produced. This emphasises the need for effective monitoring of the feed chain of food‐producing animals in order to prevent food contamination. Copyright © 2008 Society of Chemical Industry     

Aspergillus section Flavi and aflatoxins in dried figs and nuts in Algeria

The presence of Aspergillus section Flavi and aflatoxin (AF) contamination was investigated in 112 samples of peanuts, almonds and dried figs collected in Algeria. The occurrence of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in different commodities has been determined with a sensitive method based on high performance liquid chromatography (HPLC) coupled with fluorescence detection with post-column photochemical derivatisation. Analytical results indicated that 28 samples of peanuts, 16 samples of almonds and 26 samples of dried figs contained detectable levels of AFs. A total of 69 samples (61.6%) were contaminated with AFB1 ranging from the limit of quantification to 174 µg kg^?1. AFB2 was found in 12 samples (10.7%) and varied from 0.18 to 193 µg kg^?1. Seven samples revealed AF concentrations lower than the limit of quantification. Eleven peanut and fourteen dried fig samples exceeded the European maximum limits for AFB1.     

Occurrence of aflatoxins (B1, B2, G1, G2) in herbal tea consumed in Turkey

Aflatoxin contents in 12 types of herbal teas were determined by high performance liquid chromatography (HPLC) with fluorescence detector using immunoaffinity column clean-up. Forty eight samples were collected from four local herbal shops in Manisa, Turkey. Of the 48 samples analyzed, 43 were aflatoxin positive. The highest concentration of aflatoxin (34.18 µg/kg) was determined in a sample of camomile tea. The occurrence of AFB1, B2, G1 and G2 was found in samples at levels of 54, 29, 71 and 46 %, respectively. Aflatoxin B1, B2, G1 and G2 contamination levels varied from 0 to 14.2, 0 to 12.4, 0 to 13.5 and 0 to 28.7 µg/kg, respectively. Aflatoxin was not detected in five samples consisting of linseed, lime and fennel tea.     

Comparison of homogenization techniques and incidence of aflatoxin contamination in dried figs for export

To determine differences in mean aflatoxin contamination and subsample variance from dry and slurry homogenizations, 10 kg of six different, naturally contaminated dried fig samples were collected from various exporting companies in accordance with the EU Commission Directive. The samples were first dry-mixed for 5 min using a blender and sub-sampled seven times; the remainder was slurry homogenized (1 : 1, v/v) and sub-sampled seven times. Aflatoxin B₁ and total aflatoxin levels were recorded and coefficient of variations (CV) computed for all sub-samples. Only a small reduction in sub-sample variations, indicated by the lower CV values, and slight differences in mean aflatoxin B₁ and total aflatoxin levels were observed when slurry homogenization was applied. Therefore, 7326 dried figs, destined for export from Turkey to the EU and collected during the 2008 crop year, were dry-homogenized and tested for aflatoxins (B_1, B_2, G₁ and G₂) by immunoaffinity column clean-up using RP-HPLC. While 34% of the samples contained detectable levels of total aflatoxins (0.20–208.75 µg kg^?1), only 9% of them exceeded the EU limit of 4 µg kg^?1 in the range 2.0–208.75 µg kg^?1, respectively. A substantial increase in the incidence of aflatoxins was observed in 2008, most likely due to the drought stress experienced in Aydin province as occurred in 2007.     

Analysis of duplicate 24-hour diet samples for aflatoxin B1, aflatoxin M1 and ochratoxin A.

In the spring and autumn of 1994, a total diet study, in which 123 participants collected duplicates of their 24-hour diets, was carried out. The goal of this study was to determine the mass fractions of a number of analytes in these duplicate diets, so as to be able to establish oral daily intake values. After measurements were carried out for pesticides, PCBs, elements, sterols, nitrate and nitrite, and fatty acids, the duplicate diet study was concluded with analyses for aflatoxin M1, aflatoxin B1 and ochratoxin A. For this purpose a method of analysis was developed, that could simultaneously determine these mycotoxins at very low levels. The method involved chloroform extraction, liquid-liquid extraction, immunoaffinity cleanup and liquid chromatography. The method was supplemented with a procedure to confirm the identity of chromatographic peaks, assumed to represent aflatoxin M1, aflatoxin B1 and ochratoxin A. The method was in-house validated. Recoveries ranged from 68-74% for aflatoxin M1 (at spiking levels from 30-120 ng/kg, c.v. 7.6%), from 95-97% for aflatoxin B1 (at spiking levels from 50-200 ng/kg, c.v. 2.8%), and from 75-84% for ochratoxin A (at spiking levels from 150-600 ng/kg, c.v. 4.3%). Limits of quantitation (defined as signal/noise = 10) were estimated to be 24, 5 and 16 ng/kg lyophilised material for aflatoxin M1, aflatoxin B1 and ochratoxin A respectively. The newly developed method was used to analyse 123 samples of 24-hour diets. Aflatoxin M1 was detectable in 48% of the samples; the toxin contents remained below the limit of quantitation in all samples. Aflatoxin B1 could be detected in 42% of the samples; in 25% of the samples the levels were above the limit of quantitation. Ochratoxin A could be quantified in all samples. The analytical results were further processed to estimate levels of intake. Intake levels for the aflatoxins were very low, and could not reliably be established. The mean ochratoxin A intake was estimated to be 1.2 ng/kg body weight per day. This is well below the tolerable daily intake established by JECFA at 14 ng/kg body weight per day. The current dietary intake of ochratoxin A in the Netherlands is concluded to pose no appreciable health risk.     

胶体金免疫层析法快速检测黄曲霉毒素B1的研究

本文应用胶体金免疫层析技术,建立了一种快速检测食品中黄曲霉毒素B1的方法。采用柠檬酸三钠还原法制备胶体金颗粒,标记抗黄曲霉毒素B1单克隆抗体并喷于玻璃纤维上,黄曲霉毒素B1偶联抗原和二抗鼠抗驴分别结合于硝酸纤维膜上,依次将样本垫、胶金垫、硝酸纤维膜和吸水纸组装切割成胶体金试纸条并装入检测卡中。测试结果表明黄曲霉毒素B1快速检测试纸条的灵敏度为5ng/ml,检测时间为10min,批内和批间重复性为100%,假阳性率和假阴性率均为0。使用简单方便,非常适合现场快速检测黄曲霉毒素B1。     

胶体金免疫层析法检测酱油中黄曲霉毒素B1

采用胶体金免疫层析法检测酱油中的黄曲霉毒素B1。加标的酱油样品经提取后,以胶体金免疫层析法对其进行黄曲霉毒素B1测定,并与酶联免疫吸附法进行比较。结果表明,当酱油中黄曲霉毒素含量超过国家限量标准(5μg/kg),胶体金免疫层析法检测结果为阳性,说明该方法能够满足酱油样品中黄曲霉毒素B1监控的要求。     

Aflatoxins in sunflower and safflower seeds from Iran

Aflatoxin content of 173 sunflower and safflower seeds was determined by HPLC with immunoaffinity column (IAC) clean-up and fluorometric detection. Aflatoxin B₁ contamination was found in 111 samples: in 8 of the sunflower seed samples (16%) at a mean level of 40.68?ng?g^?1 and in 103 safflower seed samples (83.7%) at a mean level of 2.81?±?0.44?ng?g^?1. In 5 sunflower seed samples and 1 safflower seed sample, aflatoxin B₁ levels were higher than the maximum levels of AFB₁ under Iran regulations (5?ng?g^?1). Aflatoxin B₁ levels in 5 sunflower and 2 safflower seed samples were higher than the European Union maximum limit (2?ng?g^?1).