Aflatoxin B1 occurrence in maize sampled from Croatian farms and feed factories during 2013

The aim of the study was to determine the level of aflatoxin B₁ (AFB₁) in maize sampled from farms and feed factories situated in Northern, Central and Eastern Croatia during 2013, following the occurrence of cow milk AFM₁ contamination. Maize samples (n = 633) were analysed using Enzyme-Linked Immunosorbent Assay (ELISA) as a screening method and High Performance Liquid Chromatography Tandem Mass Spectrometry (LC–MS/MS) as a confirmatory method. Mean AFB₁ value found in maize coming from all investigated regions equalled to 81 μg/kg, with the maximal value of 2072 μg/kg found in maize obtained from Eastern Croatia. The observed contamination might have arisen on the grounds of extremely hot (>98%) and dry (<2%) weather witnessed from May to September 2012 during the maize growth and harvesting period, which might have favoured AFB₁ production and consequently the contamination of dairy cattle feeds. In order to prevent the adverse effects of AFB₁ on humans and animals, and also to reduce losses in agricultural production, systematic monitoring and further investigations of AFB₁ contamination are necessary.     

Inactivation of aflatoxigenic fungi and the reduction of aflatoxin B1 in vitro and in situ using gamma irradiation

Detailed investigation on the effect of gamma (γ) irradiation on germination, sporulation, and growth of aflatoxigenic moulds (Aspergillus parasiticus 2999, Aspergillus flavus 305, and Aspergillus niger 388), as well as on the reduction of aflatoxin B₁ (AFB₁) level in artificially and naturally contaminated maize/feed samples was performed. The results of in vitro and in situ experiments with aflatoxigenic moulds demonstrated that 5 kGy-γ irradiation manages to prevent sporulation, germination and growth of the tested moulds both when in form of a pure and when in form of a mixed culture. In the feed samples artificially contaminated with AFB₁ (50 μg kg⁻¹) 5 kGy-γ irradiation reduced AFB₁ level by around 60%, while 10 kGy-dose reduce it for around 85%. Similarly, in feed samples spiked with AFB₁ in the concentrations of 100 μg kg⁻¹ 5 kGy-dose reduced the AFB₁ level by approximately 70%, while the dose of 10 kGy reduced it by approximately 90%. The experiments on naturally contaminated maize samples (n = 30) confirmed these observations; following a 5 kGy-irradiation, the overall mean AFB₁ reduction equalled to 69.8%, while the irradiation with a 10 kGy-dose achieved the overall mean toxin reduction of 94.5%. The obtained results indicate that γ irradiation can be used to prevent the growth of aflatoxigenic moulds and to reduce the AFB₁ levels in various goods intended for animal and human consumption, thus minimizing the animal and human exposure to this carcinogenic mycotoxin.     

Natural occurrence of aflatoxin B1, ochratoxin A and citrinin in Croatian fermented meat products

When domestic animals are exposed to mycotoxins, significant amounts of the latter shall be carried over into animal products such as milk, eggs and meat. This study was carried out in order to determine the possible presence of aflatoxin B₁ (AFB₁), ochratoxin A (OTA) and citrinin (CIT) in game sausages (n = 15), semi-dry sausages (n = 25) and fermented dry-meat products (n = 50), randomly taken from individual producers and the Croatian market. AFB₁ and OTA were quantified using ELISA, while CIT was quantified using HPLC-fluorescence detector. Out of 90 samples, the fungi most frequently isolated from dry-cured meat products were of Penicillium species, while Aspergillus was isolated from only one sample. As much as 68.88% of the samples were positive for mycotoxins. Finally, the analysis of different types of meat products resulted in OTA identification in 64.44%, CIT identification in 4.44% and AFB₁ identification in 10% of the samples. The maximum OTA concentrations established in the commercial sausage samples equalled to 7.83 μg/kg, while that of AFB₁ amounted to 3.0 μg/kg. Generally, although OTA was detected in all three types of products in different percentage shares, mutual differences were not statistically significant (P > 0.05).     

Survey of aflatoxin B1 and ochratoxin A occurrence in traditional meat products coming from Croatian households and markets

The aim of this study was to investigate the occurrence of aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) in different traditional meat products circulating on Croatian markets, produced by a large number of households situated in different Croatian regions. The study involved a total of 410 samples of traditional pork meat products in terms of hams (n = 105), dry fermented sausages (n = 208), bacon (n = 62) and cooked sausages (n = 35), collected over four years period (2011–2014). Mycotoxin concentrations were quantified and confirmed using validated immunoassay method (ELISA) and high performance liquid chromatography with fluorescence detection (HPLC-FLD), respectively. The maximal observed OTA level in the fermented sausages and hams was around 5 times (5.10 μg/kg) to 10 times (9.95 μg/kg) higher than the maximal recommended level (1 μg/kg) stipulated for pork products in some EU countries. AFB₁ levels found in any given meat product analysed within this frame were not significantly higher (p > 0.05) than the applied method limit of detection. The results showed an occasional mycotoxin contamination of traditional meat products, especially that by OTA, pointing that to avoid such contamination meat and meat products on households should be produced and processed under standardised and well-controlled conditions.     

Natural mycotoxin contamination of maize (Zea mays L.) in the South region of Brazil

The natural co-occurrence of fungal metabolites in maize samples from the South region of Brazil was studied using an LC-MS/MS based multi-mycotoxin method. All maize samples (n = 148) were contaminated with fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂). Aflatoxin B₁ (AFB₁) and aflatoxin G₁ (AFG₁) were detected in 38 and 11 samples, respectively, while zearalenone (ZEN) and deoxynivalenol (DON), which were first regulated in 2014, were found in 110 and 71 samples, respectively. Apart from regulated mycotoxins, a broad range of non-regulated metabolites, from Aspergillus, Fusarium, Alternaria, Penicillium and other microbes, were also detected in maize sample. Fusarin C, a possible carcinogenic compound to humans, produced by Fusarium species and not addressed by Brazilian legislation, was detected in 54.2% of maize samples. All analysed maize samples were found to be contaminated by at least ten different metabolites, with the largest number of metabolites found in the same sample being 51.     

A survey of mycotoxins in random street-vended snacks from Lagos,Nigeria, using QuEChERS-HPLC-MS/MS

A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg⁻¹. The concentrations of aflatoxin B₁ (AFB₁) and AFB₂ reached 23 μg kg⁻¹ and 3 μg kg⁻¹, respectively. Moreover a sample of baked coconut contained α-zearalenol (α-ZOL), which was up to 54 μg kg⁻¹ in coconut candy. As considers prevalence, aflatoxins and α-ZOL were not detected in lafun and groundnut-based snacks (donkwa and kulikuli), whereas each of the three mycotoxins contaminated 12.5% (1/8) of the coconut-based samples. This is the first report of α-ZOL in cassava and coconut, and their products. AFB₁ and total aflatoxins (TAFs) concentrations exceeded the maximum allowable limit recommended by National Agency for Food and Drug Administration and Control Nigeria (NAFDAC) in one sample of baked coconut (AFB₁ = 23 μg kg⁻¹ and TAFs = 26 μg kg⁻¹) and donkwa (AFB₁ = 19 μg kg⁻¹ and TAFs = 21 μg kg⁻¹).     

Effectiveness of pulsed light treatment for degradation and detoxification of aflatoxin B1 and B2 in rough rice and rice bran

Aflatoxins primarily accumulate in the hull and bran layers of rough rice making these by-products of rice milling unsuitable for animal feed or human consumption. Contaminated rough rice is also a potential source of aflatoxin exposure to workers handling the grain during post-harvest storage and processing. Currently, no technologies are available to remove or detoxify these toxic and mutagenic fungal metabolites from contaminated rough rice. Pulsed light (PL) is a novel technology with the potential to degrade and detoxify aflatoxins in foods and their processing by-products. Rough rice was inoculated with Aspergillus flavus to produce aflatoxin B₁ (AFB₁) and B₂ (AFB₂) contamination, followed by PL treatments of 0.52 J/cm²/pulse for various durations. A PL treatment time of 80 s reduced AFB₁ and AFB₂ in rough rice by 75.0% and 39.2%, respectively; while a treatment time of 15 s reduced AFB₁ and AFB₂ in rice bran by 90.3% and 86.7%, respectively. Since PL treatments result in the degradation of aflatoxins in situ, the toxicity and mutagenic activity of the residual by-products of AFB₁ and AFB₂ after PL treatment were evaluated. Toxicity was estimated using the brine shrimp (Artemia salina) lethality assay and mutagenicity measured by the fluctuation test with Salmonella typhimurum tester strains TA98 and TA100. The mutagenic activity of AFB₁ and AFB₂ was completely eliminated by PL treatment, while the toxicity of these two aflatoxins was significantly decreased. The obtained results suggest that PL technology has a promising potential to degrade, detoxify, and inactivate the mutagenic activity of aflatoxins in rough rice and rice bran.     

Aflatoxin contamination of dried red chilies: Contrasts between the United States and Nigeria,two markets differing in regulation enforcement

Dried red chilies are among the world’s most consumed spices. From farm to fork, chilies go through cropping, harvest, drying, processing and storage. Chilies are susceptible to infection by aflatoxin producing fungi and subsequent contamination by aflatoxins at every stage. Aflatoxins are highly regulated, hepatotoxic carcinogens produced by fungi in Aspergillus section Flavi. The current study examined prevalence of aflatoxin B₁ (AFB₁) in chilies from markets across the United States (US) and Nigeria, and determined predisposition of chilies to aflatoxins post-harvest. Aflatoxin B₁ was detected in 64% chilies from US markets (n = 169), and 93% of Nigerian chilies (n = 55) with a commercial lateral flow assay (Limit of Detection = 2 μg/kg). Two percent of US samples exceeded the aflatoxin regulatory limit of 20 μg/kg, while the highest concentration detected was 94.9 μg/kg. Aspergillus spp. could be recovered only from 40% of samples from the US, and aflatoxin levels did not correlate with quantities of Aspergillus section Flavi (Colony Forming Units g⁻¹), suggesting fungi associated with chilies in US markets were killed during processing. Both average AFB₁ concentrations and fungal quantities were significantly higher (p < 0.01) in Nigerian chilies. The most contaminated sample contained 156 μg/kg AFB₁. Aflatoxin concentrations in Nigerian chilies increased as an exponential function of the quantities of Aspergillus section Flavi (r² = 0.76). Results indicate that high rates of chili consumption may be associated with unacceptable aflatoxin exposure.     

Incidence and consumer awareness of toxigenic Aspergillus section Flavi and aflatoxin B1 in peanut cake from Nigeria

Peanut cake samples were collected from major markets in five states of Nigeria and evaluated for incidence of toxigenic Aspergillus section Flavi populations, and aflatoxin B₁ (AFB₁) levels by liquid chromatography tandem mass spectrometry (LC–MS/MS). The awareness of consumers to the presence of aflatoxin in the snack and potential health risks of its regular ingestion was evaluated by questionnaire analysis. Aspergillus section Flavi populations were recovered from 83% of the peanut cake samples. Aspergillus flavus L-strain was the most predominant (>56%) across the states while Aspergillus tamarii had the least mean incidence (2.7%). The incidence of atoxigenic strains was significantly (p < 0.05) higher than that of toxigenic strains in samples from Lagos and Kaduna, while the toxigenic strains had a significantly (p < 0.05) higher incidence than the atoxigenic strains in Niger. All analyzed cake samples contained AFB₁ in concentrations exceeding the NAFDAC recommended level for AFB₁ in food and reaching up to 2824 μg/kg. There was a weak positive correlation (r = 0.32, p = 0.03) for the relationship between the incidence of toxigenic strains in the samples and AFB₁ concentration. The consumer awareness data showed that 64% of the respondents consumed peanut cake; majority of who are youth of economic and reproductive age. Eighty-five percent of the consumers lacked awareness of aflatoxin contamination in the snack and possible health risks associated with its ingestion.     

Multiple mycotoxin co-occurrence in maize grown in three agro-ecological zones of Tanzania

In this study, the co-occurrence of multiple mycotoxins in maize kernels collected from 300 households' stores in three agro-ecological zones in Tanzania was evaluated by using ultra high performance liquid chromatography/time-of-flight mass spectrometry (TOFMS) with a QuEChERS-based procedure as sample treatment. This method was validated for the analysis of the main eleven mycotoxins of health concern that can occur in maize: aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁), aflatoxin G₂ (AFG₂), ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), HT-2 toxin, T-2 toxin and zearalenone (ZEN). From each zone one major maize producing district for home consumption was chosen and 20 villages for each district were randomly selected for sampling. All mycotoxins of health concern, except for T-2 toxin, were detected in the maize samples. Particularly high levels of AFB₁ (50%; 3–1,081 μg kg⁻¹), FB₁ (73%; 16–18,184 μg kg⁻¹), FB₂ (48%; 178–38,217 μg kg⁻¹) and DON (63%; 68–2,196 μg kg⁻¹) were observed. Some samples exceeded the maximum limits set in Tanzania for aflatoxins or in European regulations for other mycotoxins in unprocessed maize. Eighty seven percent of samples were contaminated with more than one mycotoxin, with 45% of samples co-contaminated by carcinogenic mycotoxins, aflatoxins and fumonisins. Significant differences in contamination pattern were observed among the three agro-ecological zones. The high incidence and at high levels (for some) of these mycotoxins in maize may have serious implications on the health of the consumers since maize constitute the staple food of most Tanzanian population. Effective strategies targeting more than one mycotoxin are encouraged to reduce contamination of maize with mycotoxins.     

Distribution and variation of fungi and major mycotoxins in pre- and post-nature drying maize in North China Plain

Forty-four pre- and post-nature drying maize kernels were collected from North China Plain and assessed for fungi infection and mycotoxins contamination. The percentage of fungi infection was significantly higher in pre-nature drying samples than post-nature drying samples except for Fusariuim graminearum, which increases from 6.06% to 24.09%. Fusarium, Aspergillus, Alternaria and Trichoderma were main genera. Fusarium verticillioides (24.77%) and F. graminearum (15.08%) were predominant species, followed by Aspergillus niger (7.51%) and Aspergillus flavus (4.93%). FB₁ and DON were the major mycotoxins presented in the samples, followed by ZEN. All samples showed FB₁ ranging from 16.5 to 315.9 μg/kg. All post-nature drying maize kernels showed DON ranging from 5.8 to 9843.3 μg/kg, while 7 of 22 pre-nature drying samples contaminated with DON ranging from 50.7 to 776.6 μg/kg. The samples contaminated with ZEN in pre- and post-nature drying maize were 3, with the content ranging from 60.5 to 147.6 μg/kg and from 40.7 to 1056.8 μg/kg, respectively. Only 1 sample contaminated with AFB₁ of 148.4 μg/kg. The occurrence of mycotoxins is highly in accordance with the incidence of the corresponding mycotoxin-producing fungi. This is the first comprehensive comparison of fungi infection and mycotoxins contamination between pre- and post-nature drying maize kernels.     

Distribution of Salmonella clonal groups in four Brazilian feed mills

Feed is one of the major vehicles for Salmonella transmission to pigs, and its spread during the milling process may play an important role in feed contamination. Therefore, a cross-sectional study was carried out in four feed mills in order to evaluate the frequency of Salmonella isolation at various stages of feed production and to track the spread of genotypically related isolates grouped by pulsed-field gel electrophoresis (PFGE). From a total of 1269 analyzed samples, 63 (4.96%) showed the presence of Salmonella. Evaluated feed mills (A, B, C and D) presented 3.5% (n = 11/317), 1.7% (n = 5/289), 7.5% (n = 23/308) and 7.0% (n = 25/355) positive samples, respectively. Twenty-three serovars were identified, with the most frequently detected being Montevideo (n = 14, 22.2%), Anatum (n = 8, 12.7%) and Senftenberg (n = 8, 12.7%). The isolation of Salmonella was significantly higher (p = 0.002) in samples with the presence of total coliforms (36/489; 7.36%) than in the coliform-negative samples (27/780; 3.46%). Conveyors (OR = 4.43, 95% CI: 2.43–8.09) were the most likely sites of Salmonella isolation, followed by dust settled on the feed mill's floor (OR = 2.88, 95% CI: 1.41–5.33). Isolates indistinguishable on PFGE or belonging to pulsotypes with a high similarity (>95%) were identified in serovars Agona, Infantis, Montevideo, Orion, Senftenberg and Worthington. In particular, clonal groups of serovars Montevideo and Senftenberg were found to be disseminated among different sample types (ingredients, dust collected from the premise's floor and complete feed) or to be endemic in the feed mills. The dissemination of Salmonella clonal groups demonstrates the importance of control measures to avoid dust and debris accumulation on equipment surfaces.     

Determination of fungal microbiota and mycotoxins in brewers grain used in dairy cattle feeding in the State of Bahia,Brazil

The aim of this study was to determine the mycoflora and evaluate the presence of aflatoxins and ochratoxins in brewers grain used to feed dairy cattle in the State of Bahia. Twenty samples of brewers grain were collected each trimester, during a whole year, in five properties located in cities of the “recôncavo baiano” (Bahia, Brazil) for a total of 80 samples. Samples were analyzed for aflatoxins and ochratoxins by fluorimetry with immunoaffinity columns. Aspergillus was the most frequently isolated genus (42.5%), followed by Penicillium, Mucor, Rhizopus and Fusarium. Mycotoxicological analyses did not show the presence of ochratoxins, but the presence of aflatoxins was observed in 33.75% (27/80) of the samples, with contamination levels between 1 and 3 μg/kg.     

Isolation and characterization of a Bacillus subtilis strain with aflatoxin B1 biodegradation capability

Aflatoxins are type of mycotoxins mainly produced by Aspergillus flavus and a common contaminant of food and grain, posing a serious economic and health problem worldwide. In order to find efficient bacteria to remove or detoxify these mycotoxins, a bacterial strain capable of degrading aflatoxin B₁ (AFB₁) was isolated from soil samples using a culture medium containing coumarin as the sole carbon source. Based on 16S rRNA gene sequence analysis, this isolate was identified as Bacillus subtilis JSW-1; its further characterization showed that it could inhibit the growth of A. flavus with an inhibition ratio of 58.3% and could degrade AFB₁ by 67.2% after incubation at 30 °C for 72 h. The aflatoxin B₁-degrading activity of isolate JSW-1 was predominantly attributed to the cell-free supernatant and this activity was found to be heat stable but sensitive to proteinase K treatment, indicating that the extracellular proteins or enzymes are responsible for the AFB₁ degradation. In addition, no degradation products of AFB₁ could be detected by liquid chromatography-mass spectrometry (LC-MS) analysis, indicating that the parent AFB₁ might be biotransformed to compounds with chemical properties different from that of AFB₁.     

Aflatoxins and fumonisins in rice and maize staple cereals in Northern Vietnam and dietary exposure in different ethnic groups

Mycotoxins in food are increasingly a food safety hazard concern in particular in developing countries. This study was performed to determine the occurrence and determinants of aflatoxin and fumonisin contamination in rice and maize and to assess health risks through dietary intake exposure among ethnic minority groups in northern Vietnam. A total of 111 rice and 102 maize samples, were tested for occurrence of fungi and mycotoxins, i.e. aflatoxins (AF’s) and fuminisin B (FB). Results showed that 107 (96.4%) rice and 84 (82.4%) maize samples were contaminated by fungi. Aspergillus flavus was found in 68 (61.3%) rice and 30 (29.4%) maize samples, Aspergilus parasiticus in 40 (36.0%) rice and 27 (26.7%) maize samples. AF’s - were detected in 27 rice (24.3%) and 27 maize samples (26.4%) at minimum and maximum levels in rice of 2.06 and 77.8 ng/g and 20.5 and 110 ng/g in maize, respectively. Nine (8.1%) rice and 24 (23.5%) maize samples contained FB at ranges of 2.3–624 ng/g in rice and 5.6–89.8 ng/g in maize. Data collected through interviews and observations in households showed that type of crop, storage duration and presence of fungi, particularly mycotoxigenic fungi were important risk factors for AF’s and FB contamination. Based on daily food consumption data, the estimated average exposure dose of aflatoxin B₁(AFB₁) from rice was 21.7 ng/kg bw/day for adults and 33.7 ng/kg bw/day for children. For FB, the rice based average exposure amounted to 536 ng/kg bw/day for adults and 1019 ng/kg bw/day for children. The calculated excess risk of liver cancer incidence by ingestion of cereals containing AFB₁ was 1.5 per 100,000 adults and 2.3 per 100,000 children per year. The average intake of FB was calculated to be lower than the tolerable diet intake (TDI). Our findings highlight that rice and maize are contaminated with mycotoxins at levels representing actual health hazards for the ethnic minority groups consuming these stable cereals. Proper drying and storage conditions in households are likely to reduce the mycotoxin contamination.     

Aflatoxin M1 in milk and traditional dairy products from west part of Iran: Occurrence and seasonal variation with an emphasis on risk assessment of human exposure

In the present study, a total of 358 samples consisting of raw milk of cow (n = 64), goat (n = 56) and sheep (n = 52); traditional cheese (n = 40), yoghurt (n = 42), Kashk (n = 40), Doogh (n = 44) and Tarkhineh (n = 20) were analyzed for aflatoxin M₁ (AFM₁) by using an enzyme linked immunosorbent assay (ELISA). Frequency of AFM₁ and its concentration ranges in the ELISA positive samples were determined by high performance liquid chromatography with fluorescence detection (HPLC-FD). AFM₁ contamination was 84.3%, 44.6% and 65.3% for cow, goat and sheep raw milks, respectively. Moreover, AFM₁ was in 65.5%, 23.8%, 14%, 13.6% and 35.0% of cheese, yoghurt, Kashk, Doogh and Tarkhineh samples, respectively. Percentages of cow milk, goat milk, sheep milk and cheese samples exceeding the EU limit were 35.9%, 11.1%, 26.9% and 10%, respectively. HPLC analyses confirmed the ELISA results although the percentages of AFM₁ contamination in raw milk and dairy products were lower than that of ELISA. There were significant differences (P < 0.05) between the mean AFM₁ contents of raw milk, cheese and yoghurt samples during winter and summer seasons. Our study demonstrated that there is a potential risk for liver cancer due to the consumption of milk and dairy products in Iranian consumers.     

Fumonisins in brewers grain (barley) used as dairy cattle feed in the State of Bahia,Brazil

Fumonisins are mycotoxins produced by the genus Fusarium that may induce toxic effects in several animal species and may be found in several kinds of foods and feed. In the State of Bahia, Brazil, brewers grain, which are a brewery by-product, have been largely used in the feeding of animals, specially dairy cattle, due to their nutritional value and low cost of transportation. The aim of this study was to establish the presence of fumonisins in brewers grain used as dairy cattle feed in the State of Bahia. Twenty samples of brewers grain were collected every three months during a whole year, for a total of 80 samples, in five properties located in the “reconcavo baiano”. These samples were analyzed for the presence of fumonisins using high efficiency liquid chromatography (HPLC). Results showed contamination of 58 (72.5%) samples, with contamination mean level equal to 226.5 μg/kg, with 50.30 and 908.47 μg/kg as the minimum and maximum levels, respectively. This is the first report of the occurrence of this mycotoxin in the State of Bahia.     

Development of a new broad-specific monoclonal antibody with uniform affinity for aflatoxins and magnetic beads-based enzymatic immunoassay

To reduce the incidence of false-positive and false-negative results caused by high or low cross-reactivity (CR%) values of the antibodies for total aflatoxins (AFs, AFB₁+AFB₂+AFG₁+AFG₂) detection, a new broad-specific monoclonal antibody (MAb) with uniform affinity, named 5H3, was developed. Moreover, magnetic beads (MBs) replaced microplates as immobile phase to improve the sensitivity of the enzymatic immunoassay. Then, a direct competitive enzyme-linked immunosorbent assay (ELISA) based on MBs (MBs-dcELISA) that could simultaneously detect the total AFs with similar sensitivity was developed. Following optimization of conditions, the half maximal inhibitory concentrations (IC₅₀) of the MBs-dcELISA in buffer were 0.05 ng/mL for AFB₁, 0.04 ng/mL for AFB₂, 0.05 ng/mL for AFG₁, 0.06 ng/mL for AFG₂. The corresponding CR% values were 100%, 125%, 100% and 83.3%, respectively. The limit of detection (LOD) of the MBs-dcELISA for the total AFs was 0.21 ng/g with a working range from 0.22 ng/g to 19.8 ng/g, and the recoveries for the total AFs ranged from 74.5% to 96.5% with coefficients of variation (CV) under 12.1% in spiked maize samples. In addition, the MBs-dcELISA was more sensitive than the conventional dcELISA. Finally, the MBs-dcELISA was applied to screen 9 naturally contaminated maize samples and 6 spiked samples and the results indicated a good agreement with that obtain by HPLC-MS/MS method.     

Degradation and detoxification of AFB1 by Staphylocococcus warneri,Sporosarcina sp. and Lysinibacillus fusiformis

Effects associated with aflatoxins (AFs), principally aflatoxin B₁ (AFB₁) have necessitated strategies to eliminate their occurrence in commodities along the food chain. This study therefore, investigated the AFB₁ biodegradation ability of Staphylococcus warneri, Sporosarcina sp. and Lysinibacillus fusiformis liquid cultures and cell lysates (disrupted in the presence or absence of protease inhibitors to obtain lysates). These were incubated with AFB₁ (2.5 μg/mL) for 3, 6, 12, 24 and 48 h. AFB₁ degradation was subsequently monitored on high performance liquid chromatography (HPLC) and results indicated that after 48 h, % AFB₁ degradation by the liquid cultures of Lysinibacillus fusisormis, S. warneri and Sporosarcina sp. were 61.3, 47.7 and 46.9%, respectively. After 12 h of incubation, a 100% AFB₁ degradation was observed for all protease inhibited lysates tested. To establish toxicity of the AFB₁ biotransformed products, results from a cytotoxicity study against human lymphocytes demonstrated that the products exhibited significantly (p ≤ 0.05) lower cytotoxic effect compared to the parent AFB₁. From this study, it can be deduced that the mechanism of AFB₁ degradation was enzymatic and that protease inhibition of cells before disruption, could increase this enzymatic activity. Conclusively, the potential of these lysates as a biotechnological approach towards decontaminating AFB₁ is promising.     

Identification of aflatoxin B1 on maize kernel surfaces using hyperspectral imaging

A shortwave infrared (SWIR) hyperspectral imaging system with wavelength range between 1000 and 2500 nm was used to assess the potential to detect aflatoxin B₁ (AFB₁) contaminants on the surface of healthy maize kernels. Four different AFB₁ solutions were prepared and deposited on kernels surface to achieve 10, 20, 100, and 500 ppb, respectively. A drop of 20% methanol was dipped on the surface of 30 healthy kernels in the same way to generate the control samples. Based on the standard normal variate (SNV) transformation spectra, principal components analysis (PCA) was used to reduce the dimensionality of the spectral data, and then stepwise factorial discriminant analysis (FDA) was performed on latent variables provided by the PCA's. Furthermore, beta coefficients of the first three of four discriminant factors were analyzed and key wavelengths, which can represent AFB₁ and be used to differentiate different level of AFB₁ were indentified. Furthermore, 150 independent samples were used as verification set to test the reproducibility of the proposed method. A minimum classification accuracy of 88% was achieved for the validation set and verification set. Results indicated that hyperspectral imaging technology, accompanied by the PCA-FDA method, can be used to detect AFB₁ at concentrations as low as 10 ppb when applied directly on the maize surface.