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.     

Occurrence and co-occurrence of Fusarium mycotoxins in wheat grains and wheat flour from Romania

In this study, the presence of fourteen Fusarium mycotoxins, legislated by the European Union – deoxynivalenol, zearalenone, HT-2 and T-2 toxins (EC/1881/2006; 2013/165/EU), or non-legislated (five trichothecens and five “emerging” mycotoxins), was evaluated in 31 whole unprocessed wheat samples and 35 white wheat flour samples from different areas of Romania. For this purpose, a validated multi-mycotoxins liquid chromatography tandem mass spectrometry method was applied. Seventy three percent of the analyzed samples contained at least one mycotoxin. The highest occurrence was for enniatin B, 71% of the analyzed samples being positive (21–407 μg kg⁻¹). Regarding the legislated mycotoxins, deoxynivalenol was detected in 14% (111–1787 μg kg⁻¹) of the samples, while zearalenone was detected in 9% (51–1135 μg kg⁻¹). Only one sample was positive for neosolaniol. Concerning the co-occurrence, 42% of the samples were contaminated with two to five mycotoxins, the most frequent being the binary or tertiary combinations of enniatins. This is the first study applied to Romanian wheat grains and flour samples using a high sensitive multi-mycotoxins method, and which included also “emerging” mycotoxins.     

Aflatoxins and ochratoxin A in dried eggplant and green bell pepper

In this study, 50 dried eggplant and 50 dried green bell pepper samples were analyzed in terms of their aflatoxin and ochratoxin A (OTA) content. Aflatoxins G₂, G₁, B₂, and B₁, and OTA contents were analyzed using high-performance liquid chromatography with a flame ionization detector (HPLC–FID). Total aflatoxin and, as well as aflatoxin G₂, G₁, B₂, and B₁ content in dried eggplant samples were ranged between 0.82 and 2.58, 0.10–0.23, 0.32–1.35, 0.12–0.67, and 0.17–0.71 μg kg⁻¹, respectively. Total aflatoxin and, as well as aflatoxin G₂, G₁, B₂ and B₁ content in dried green bell pepper samples were 0.81–2.42, 0.11–0.22, 0.32–1.38, 0.13–0.66, and 0.18–0.91 μg kg⁻¹, respectively. OTA content was varied from 8.88 to 21.35 μg kg⁻¹ in eggplant samples, and from 15.38 to 24.70 μg kg⁻¹ in dried green bell pepper samples. Of the dried eggplant samples and dried green bell pepper samples, 36% and 24% of them, respectively, had aflatoxin B₁ values which were below the minimum limit of detection (LOD) of 0.05 μg kg⁻¹. None of the analyzed samples exceeded the legal limit values of 10 μg kg⁻¹ for total aflatoxin content, and 5 μg kg⁻¹ for aflatoxin B₁ content. However, 80% of the dried eggplant samples and 100% of the dried green bell pepper samples exceeded the legal limit value for OTA content (15 μg kg⁻¹). According to the results, it was concluded that dried vegetables should be examined in terms of their aflatoxins. It is essential to analyze OTA content more thoroughly, as it has the potential to pose a risk for public health, as well as for the economy.     

Multimycotoxin analysis of sorghum (Sorghum bicolor L. Moench) and finger millet (Eleusine coracana L. Garten) from Ethiopia

The natural contamination of sorghum and finger millet by toxigenic fungi and associated mycotoxins has been studied. All the tested sorghum and finger millet samples were found to be contaminated by Fusarium and Aspergillus species. Sorghum was considerably more likely to be contaminated by both genera than finger millet. Penicillium, Alternaria, Rhizopus and Epicoccum species were also present in both grains albeit at lower frequencies. Multimycotoxin analysis using LC–MS/MS revealed the contamination of sorghum and finger millet by 84 and 62 metabolites, respectively. The prevalence of major mycotoxins was lower than 15% in sorghum except zearalenone that occurred in one third of the samples at average level of 44 μg/kg. In finger millet major mycotoxins occurred at a prevalence of 6–52% with zearalenone being the dominant and occurring at average level of 76 μg/kg. Aflatoxins B₁, B₂, G₁, G₂ and M₁ were detected in at least one sorghum sample while only aflatoxins B₁ and G₁ were present in finger millet samples. The average aflatoxins B₁ and G₁ concentrations in sorghum have been higher than European standards. But the level of B₂, G₂ and M₁ in sorghum and that of B₁ and G₁ in finger millet have been lower. Apart from aflatoxin precursors and other Fusarium metabolites, a broad range of additional metabolites were detected in sorghum and finger millet.     

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.     

Aflatoxin contamination in unrecorded beers from Kenya – A health risk beyond ethanol

Samples of unrecorded opaque beers (n = 58; 40 based on maize, 5 on sorghum and 13 on other plants) and recorded wines (n = 8) in Kenya were screened for aflatoxins using a rapid ELISA technique followed by confirmation using liquid chromatography-tandem mass spectrometry. Six of the maize beers were obtained from Kibera slums in Nairobi County. Aflatoxin contamination was detected in six unrecorded beers (10%), but in none of the recorded wines. Remarkably, three of the aflatoxin positive samples were from the Kibera slums.The mean concentration of aflatoxins in the positive samples was 3.5 μg/L (range 1.8–6.8 μg/L), corresponding for an average consumption of 500 mL (1 standard drink) to a margin of exposure (MOE) of 36 (range: 15–58), which is considered as ‘risk’. On the other hand, the alcoholic strength of the aflatoxin positive samples had a mean of 4.3% vol (range 3.5–4.8%) corresponding to a MOE of 2.5 (range of 2.2–3.0) for the equivalent consumption volume. While aflatoxins pose a risk to the consumer, this risk is about 10 times lower than the risk of ethanol.The Joint FAO/WHO Expert Committee on Food Additives sets no acceptable daily intake for aflatoxins since they are genotoxic carcinogens and instead recommends for the reduction of aflatoxin dietary exposure as an important public health goal, particularly in populations who consume high levels of any potentially aflatoxins-contaminated food. Nevertheless, ethanol still posed a considerably higher risk in the unrecorded beers examined. However, consumers should be informed about aflatoxins, as these are an involuntary and unknown risk to them. In addition, producers should be educated about measures to reduce aflatoxins in alcoholic beverages.     

Natural occurrence of mycotoxins in cereals and spices commercialized in Morocco

Sixty samples of cereals (20 of corn, 20 of barley, and 20 of wheat) and 55 samples of spices (14 of paprika, 12 of ginger, 14 of cumin, and 15 of pepper) purchased from popular markets of Rabat and Salé in Morocco were analyzed for mycotoxins.Cereals samples were all analyzed for ochratoxin A (OTA). The average levels of contamination were 1.08, 0.42, and 0.17 μg/kg for corn, wheat, and barley, respectively. Samples of corn were also analyzed for zearalenone (ZEA) and fumonisin B₁ (FB₁) the average contaminations were 14 and 1930 μg/kg, respectively. Co-occurrence of OTA, FB₁, and ZEA was also checked. Spices samples were analyzed only for aflatoxins (AFs) and the average contaminations found for aflatoxin B₁ (AFB₁) were 0.09, 0.63, 2.88 and 0.03 μg/kg for black pepper, ginger, red paprika and cumin, respectively. The higher level of contamination was found in red paprika (9.68 μg/kg).The present report is the first one ever drafted on the natural co-occurrence of OTA, FB₁ and ZEA in cereals and on the occurrence of AFs in spices from Morocco.     

Fast and sensitive aflatoxin B1 and total aflatoxins ELISAs for analysis of peanuts,maize and feed ingredients

Aflatoxins are a group of carcinogenic compounds produced by Aspergillus fungi that can grow on different agricultural crops. Both acute and chronic exposure to these mycotoxins can cause serious illness. Due to the high occurrence of aflatoxins in crops worldwide fast and cost-effective analytical methods are required for the identification of contaminated agricultural commodities before they are processed into final products and placed on the market. In order to provide new tools for aflatoxin screening two prototype fast ELISA methods: one for the detection of aflatoxin B₁ and the other for total aflatoxins were developed. Seven monoclonal antibodies with unique high sensitivity and at the same time good cross-reactivity profiles were produced. The monoclonal antibodies were characterized and two antibodies showing IC₅₀ of 0.037 ng/mL and 0.031 ng/mL for aflatoxin B₁ were applied in simple and fast direct competitive ELISA tests. The methods were validated for peanut matrix as this crop is one of the most affected by aflatoxin contamination. The detection capabilities of aflatoxin B₁ and total aflatoxins ELISAs were 0.4 μg/kg and 0.3 μg/kg for aflatoxin B₁, respectively, which are one of the lowest reported values. Total aflatoxins ELISA was also validated for the detection of aflatoxins B₂, G₁ and G₂. The application of the developed tests was demonstrated by screening 32 peanut samples collected from the UK retailers. Total aflatoxins ELISA was further applied to analyse naturally contaminated maize porridge and distiller's dried grain with solubles samples and the results were correlated with these obtained by UHPLC-MS/MS method.     

Three years of monitoring of PCDD/F,DL-PCB and NDL-PCB residues in bovine milk from Lombardy and Emilia Romagna regions (Italy): Contamination levels and human exposure assessment

In the three-years period 2012–2014, 160 cow milk samples from farms located in Lombardy and Emilia Romagna regions (Italy) were analyzed during the implementation of the Italian National Residues Monitoring Plan to assess the presence of PCDD/F, DL-PCB and NDL-PCB residues. The obtained contamination data were combined with cow milk consumption data from the Italian national dietary survey to estimate PCDD/F, DL-PCB and NDL-PCB human dietary exposure through the consumption of whole, semi skimmed and skimmed bovine milk. The exposure assessment was carried out separately for children, teenagers, adults and elderly. Average contamination levels of the analyzed samples were found to be 1.26 pg WHO-TEQ/g fat for the sum of PCDD/Fs and DL-PCBs and 9.30 ng/g fat for the sum of the 6 NDL-PCB indicators. PCB 126 was found to be the main contributor to the total WHO-TEQ. Using the upper bound approach, the estimated mean dietary intakes ranged from 0.07 pg WHO-TEQ/kg bw per day to 0.39 pg WHO-TEQ/kg bw per day for the sum of PCDD/Fs and DL-PCBs and considering exposure from whole milk. NDL-PCB mean dietary intakes resulted between 0.52 ng/kg bw per day and 2.86 ng/kg bw per day for consumption of whole milk. Children and teenagers were found to be the most exposed groups. This is the first time that Italian consumers exposure to NDL-PCBs is assessed using contamination data of cow milk produced in Italy.     

Monitoring of selected aflatoxins in brewing materials and beer by liquid chromatography/mass spectrometry

In 2008-2011 a total set of 333 samples of brewing raw materials and beer were analyzed for the presence of aflatoxins B₁, B₂, G₁ and G₂. The standard analytical method using high-performance liquid chromatography coupled to mass spectrometric detection with immunoaffinity column clean-up was applied. The method was validated. Limits of detection varied from 0.04 to 0.12 μg/kg in barley and malt, 0.08-0.58 μg/kg in different hop samples, 0.04-0.12 μg/kg in brewers’ yeast and spent grains and 1.5-4.7 ng/l in beer. Limits of quantification varied from 0.13 to 0.39 μg/kg in barley and malt, 0.25-1.94 μg/kg in different hop samples, 0.13-0.39 μg/kg in brewers’ yeast and spent grains and 5.1-15.2 ng/l in beer. In 7 of 216 samples of brewing raw materials (3.2%), aflatoxins were found at trace concentrations to 1.2 μg/kg. In 6 of 117 (5.1%) beer samples, aflatoxins were detected at concentrations to 31.0 ng/l. Values in barley and malt did not exceed the maximum allowable limit set by the European Union.     

Development and validation of an accurate and rapid LC-ESI-MS/MS method for the simultaneous quantification of aflatoxin B1, B2, G1 and G2 in lotus seeds

An accurate and rapid LC-ESI-MS/MS analytical method was developed and validated for the simultaneous determination of aflatoxin B₁, B₂, G₁ and G₂ in lotus seeds. The samples were firstly extracted with methanol-water solution (80:20, v/v), and then cleaned up by immunoaffinity columns. The mass spectrometer was operated in the positive ionization electrospray (ESI+) mode using multiple reaction monitoring (MRM) for analysis of four aflatoxins. The transitions of m/z 313 → 285 (AFB₁, CE 33 eV), m/z 315 → 259 (AFB₂, CE 37 eV), m/z 329 → 243 (AFG₁, CE 37 eV) and m/z 331 → 257 (AFG₂, CE 37 eV) were used to quantify these four aflatoxins, respectively. The limits of detection (LODs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.007, 0.005, 0.003 and 0.005 μg kg^?1 based on a signal-to-noise ratio of 3:1, respectively. The limits of quantification (LOQs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.02, 0.015, 0.01 and 0.015 μg kg^?1 based on a signal-to-noise ratio of 10:1, respectively. Recoveries for samples of spiked lotus seeds were all above 66% with relative standard deviation all below 15% for all compounds. Nineteen out of twenty batches of lotus seeds collected from different drug stores or markets in China were found to be contaminated with aflatoxins at different levels ranging from 0.02 to 688.4 μg kg^?1.     

Determination of multi-mycotoxin occurrence in maize based porridges from selected regions of Tanzania by liquid chromatography tandem mass spectrometry (LC-MS/MS), a longitudinal study

Residents of certain areas of Tanzania are exposed to mycotoxins through the consumption of contaminated maize based foods. In this study, 101 maize based porridge samples were collected from villages of Nyabula, Kikelelwa and Kigwa located in different agro-ecological zones of Tanzania. The samples were collected at three time points (time point 1, during maize harvest; time point 2, 6 months after harvest; time point 3, 12 months after harvest) over a 1-year period. Ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used to detect and quantify 9 mycotoxins: aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁), aflatoxin G₂ (AFG₂), fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), deoxynivalenol (DON), ochratoxin A (OTA) and zearaleneone (ZEN) in the samples following a QuEChERS extraction method. Eighty two percent of samples were co-contaminated with more than one group of mycotoxins. Fumonisins (FB₁ + FB₂) had the highest percentage occurrence in all 101 samples (100%) whereas OTA had the lowest (5%). For all three villages the mean concentration of FB₁ was lowest in samples taken from time point 2. Conversely, In Kigwa village there was a distinct trend that AFB₁ mean concentration was highest in samples taken from time point 2. DON concentration did not differ greatly between time points but the percentage occurrence varied between villages, most notably in Kigwa where 0% of samples tested positive. ZEN occurrence and mean concentration was highest in Kikelelwa. The results suggest that mycotoxin contamination in maize can vary based on season and agro-ecological zones. The high occurrence of multiple mycotoxins found in maize porridge, a common weaning food in Tanzania, presents a potential increase in the risk of exposure and significant health implications in children.     

Method development for the analysis of phthalate esters in tea beverages by ionic liquid hollow fibre liquid-phase microextraction and liquid chromatographic detection

A novel method, termed ionic liquid-based hollow fibre liquid phase microextraction (IL-HF-PLME), coupled with high-performance liquid chromatography (HPLC) was developed for separation and preconcentration of three phthalate esters (PAEs) in tea beverage. In the present study, ionic liquid 1-butyl-3-methy-limidazolium hexafluorophosphate ([BMIm]PF₆) was placed in the porous-walled polypropylene hollow fibre as the acceptor phase, and nonanol was used as the supported liquid membrane phase that accomplished extraction. Several important parameters influencing the extraction efficiency were investigated in detail. Under the optimized conditions, good linearity occurred in the range of 5–1000 ng mL⁻¹ with the correlation coefficients values above 0.998. The limits of detection ranged from 0.67 to 1.73 ng mL⁻¹. Recoveries of three PAEs in two kinds of spiked tea beverage samples (PAEs, 10.0–100.0 ng mL) were between 94.2 and 103.4%, with relative standard deviations (RSDs) ranged from 1.77 to 3.02%. The enrichment factors were 200. The developed IL-HF-PLME method allowed the simple, rapid, and sensitive determination of phthalate esters in tea beverage samples with an extraction time of just 4 min.     

Safety of a street vended traditional maize beverage,ice-kenkey,in Ghana

Ice-kenkey is a chilled cereal beverage sold as street food in some open markets in Ghana. It is produced by mashing and sweetening kenkey, a stiff dumpling produced from fermented maize meal. The safety of street vended ice-kenkey was assessed by microbiological, elemental and myco-toxicological analysis of ice-kenkey and intermediary products obtained from 16 producers in four open markets in the Accra and Tema metropolis. A tenfold increase in counts of aerobic mesophiles, and yeast and moulds were recorded during the production of ice-kenkey. Coliform bacteria, E. coli and Staphylococcus aureus which were not detected in the starting materials were found partway through production or in the final product. The mean microbial counts in the packaged ice-kenkey were 10⁶–10⁷ CFU/g for aerobic mesophiles, 10⁴–10⁵ CFU/g for yeast and moulds, 10–1000 CFU/g for total coliforms and 10–100 CFU/g for S. aureus. E. coli counts of 10 CFU/g were recorded in samples from three out of the four markets. The microbial load could be eliminated by pasteurizing ice-kenkey at 80 °C for 15 min. The mean concentration in mg/kg of Fe was between 15.97 and 29.48, Cu, 0.57 to 1.41, Mn, 0 to 2.55, Pb 0 to 1.25 and Zn 0.47 to 6.17. Total aflatoxins content in samples ranged from 7.04 to 22.17 μg/kg and included a range of 7.01–20.54 for aflatoxin B₁, 0.51 to 1.63 for aflatoxin B₂ and 0–0.47 μg/kg for aflatoxin G_I. Aflatoxin G₂ was not detected in any of the samples. A simplified training module based GMP, GHP and a HACCP plan was developed and used to train ice-kenkey producers in Accra in collaboration with municipal authorities.     

Effect of nutmeg (Myristica fragrans) essential oil on the oxidative and microbial stability of cooked sausage during refrigerated storage

This paper examines the oxidative and microbial stability of cooked sausages, produced with the addition of 10 ppm (NO1) and 20 ppm (NO2) nutmeg (Myristica fragrans) essential oil. Instrumental parameters of color (CIE L*, CIE a* and CIE b*), Thiobarbituric acid-reactive substance (TBARS) values, microbial profile and sensory properties of aroma have been determined on the 1st, 30th, 45th, and 60th day of storage. Addition of the nutmeg essential oil had no effect on the color of cooked sausages. At the end of the storage, NO2 sausages had the best oxidative and microbial stability. TBARS values of NO1 and NO2 sausages were 1.21 mg MDA/kg and 0.95 mg MDA/kg, respectively, and were significantly lower (P < 0.05) compared to control (1.53 mg MDA/kg). Total number of aerobic mesophilic bacteria was lowest in NO2 sausages (78.3 cfu/g) and highest in control (185 cfu/g). After 45 and 60 days of storage, sensory properties of aroma of NO1 (4.21; 3.92) and NO2 sausages (4.39, 4.28) were better compared to those in control (4.07, 3.25). Hence, the addition of nutmeg essential oil in amount of 20 ppm can be successfully applied in order to extend the shelf life of cooked sausages.     

Mycotoxins occurrence in Argentina’s maize (Zea mays L.), from 1999 to 2010

The natural occurrence of mycotoxins in maize in Argentina, from 1999 to 2010 was analysed. Total aflatoxins, deoxynivalenol and zearalenone, were detected and quantified by TLC. Each aflatoxins and fumonisins was quantified by HPLC. A total of 3246 samples for freshly harvested (1655) and storage (1591) was obtained from different regions, from 1999 to 2010. Except for 2003 year in harvest, and for 2007 in storage, aflatoxins levels were low. Average values of aflatoxin B₁ for freshly harvested samples were between 0.38 and 2.54 μg kg⁻¹ and for storage samples were between 0.22 and 4.5 μg kg⁻¹. The average values and frequency of contamination for zearalenone and deoxynivalenol, were low for all years. The average zearalenone contamination in samples of freshly harvested, showed values from no detected up to 83 μg kg⁻¹, and in storage samples, from no detected to 17 μg kg⁻¹. The average deoxynivalenol contamination of freshly harvested, showed values from no detected up to 140 μg kg⁻¹, and for storage samples showed values from no detected to 14 μg kg⁻¹. The percentage of maize samples contaminated by fumonisins was between 90 and 100% for all years studied; the average levels were from 1773 to 9093 μg kg⁻¹ for freshly harvested maize and for storage from 2525 to 11,528 μg kg⁻¹.Co-occurrence of aflatoxins and fumonisin was the most frequent (8.4%), followed by zearalenone and fumonisins (2%).     

Determination of melamine in milk using surface plasma effect of aggregated Au@SiO2 nanoparticles by SERS technique

A simple and rapid method to detect melamine in liquid milk by Surface-enhanced Raman Scattering (SERS) technique was presented. The pretreatment procedure of milk samples only contains hydrochloric acid treatment and twice centrifugation. In order to reduce the distortion about the Raman signals originating from charge transfer and electronic tunnelling effect, SiO₂ shell-isolated gold nanoparticles (Au@SiO₂ NPs) instead of Au NPs were employed to enhance signal intensity. Aggregation occurs when the Au@SiO₂ NPs colloid is mixed with the melamine solution or the treated milk containing melamine. Different from aggregated Au NPs, these aggregated Au@SiO₂ NPs on Cu substrate can undistortedly enhance the Raman signals of melamine using surface plasma effect. Quantitative analysis was tried and the results showed a good linearity (R² ≈ 0.99) when the melamine concentration was between 0.5 mgL⁻¹ and 5 mgL⁻¹. The detection sensitivity satisfies the requirement of the Codex Alimentarius Commission and this method can be practically used for melamine detection in milk.     

Fungal contamination and aflatoxin B1 of ‘egusi’ melon seeds in Nigeria

One hundred and thirty seven samples of melon seeds (Colocynthis citrullus L.) from randomly selected farmers’ stores in the humid forest and Northern Guinea savanna of Nigeria were analysed for the incidence of diseased seeds, moisture content, associated moulds and levels of aflatoxin B₁ contamination. The proportion of diseased seeds ranged from 2.5 to 37.3% in the forest and 2.1 to 17.9% in the savanna, while the seed moisture content varied from 5.3 to 10.4%, and 4.6 to 9.5% respectively. All the samples contained moulds, with the two genera, Aspergillus and Penicillium predominating, while A. flavus had the highest species count. The other common fungal isolates in order of decreasing incidence were A. niger, P. citrinum, Botryodiplodia theobromae, Cladosporium sp and A. clavatus. Thin layer chromatography analysis showed that 32% in the forest and 21% samples in the savanna contained aflatoxin B₁ with mean levels of 14.8 μg/kg in the forest and 11.3 μg/kg in the savanna respectively. Significant positive correlations were found between number of aflatoxin B₁ positive samples and the percentage of A. flavus infected samples and between the levels of diseased seeds and the levels of aflatoxin B₁ contamination.     

Combination of nisin and ε-polylysine with chitosan coating inhibits the white blush of fresh-cut carrots

Effects of the combination of nisin and ε-polylysine with chitosan coating on quality maintenance and white blush inhibition were investigated in fresh-cut carrots. Fresh-cut carrots were treated with 1% lactic acid solution (v/v), 1% chitosan solution (w/v), or 1% chitosan solution containing 64 μg/mL nisin and 250 μg/mL ε-polylysine (LA + CH + Nisin + ε-PL). The samples were packed in polyethylene plastic bags and stored at 4 °C for 9 days. Changes in sensory attributes, physicochemical indices, respiration rate, microbiological counts and white blush were measured. Results showed that LA + CH + Nisin + ε-PL significantly (P < 0.05) inhibited respiration rate, decline of ascorbic acid and growth of microorganism (yeast and mold, total viable counts, total coliforms counts, Staphylococcus aureus and Pseudomonas spp.), and increased total phenol content and phenylalanine ammonialyse (PAL) activity compared with the control after 9-day storage. It was also strongly effective in inhibiting the white blush of fresh-cut carrots. Furthermore, LA + CH + Nisin + ε-PL significantly (P < 0.05) reduced the lignin synthesis in fresh-cut carrots by inhibiting the cinnamate-4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL) activity, as well as Dc4CL and DcC4H gene expression. Our results may provide some basis for the use of the combination of nisin and ε-polylysine with chitosan coating as an alternative preservation method for fresh-cut carrots.     

Mycoflora and absence of aflatoxin contamination of commercialized cassava chips in Benin, West Africa

Studies conducted in Benin, in which the main staple foods are maize, cassava, groundnuts and yams, showed high levels of aflatoxin residues in blood of the exposed population. The natural contamination with fungi and aflatoxins in cassava chips sold at markets in Benin, West Africa was investigated. A total of sixty samples were sampled from open markets in 11 districts of 3 agroecological zones and analyzed for the presence of mycoflora and aflatoxin B₁, B₂, G₁ and G₂. Fourteen genera of fungi were associated with marketed dried cassava chips. Within these, twenty- two isolates were identified to species level, whereas four were identified only to genus. The dominating fungal species isolated were Rhizopus oryzae, Nigrospora oryzae, Chrysonilia sitophila, Cladosporium resinae, Cladosporium herbarum, Apergillus niger and Aspergillus flavus. Fifty-four out of sixty samples were contaminated with A. flavus. The rate of occurrence in CFU/g of A. flavus fungi was lower than for all other fungal species together. Aflatoxin was not detected in any of the samples analyzed using HPLC with post-column photochemical derivatization and fluorescence detection. The limit of detection (LOD) was 0.1 μg/kg. Results from this study suggest cassava chips are unlikely to be a source of aflatoxin in Benin, and that other staples such as maize and groundnuts are more important in aflatoxin exposure. Therefore it can be speculated that staples like maize and groundnut are more important in aflatoxin exposure.