Phytochemical screening, antioxidant activity of selected methanolic plant extracts and their detoxification capabilities against AFB1 toxicity.

Aflatoxin B1 (AFB1) is a secondary metabolite produced principally by Aspergillus parasiticus and A. flavus. It is one of the most potent and commonly occurring dietary carcinogen with its carcinogenic potential being linked to the formation of DNA adducts and reactive oxygen species (ROS). Plant extracts contain a plethora of biologically active phytochemicals that act against ROS. This study aimed to assess the phytochemical content and antioxidant activity of methanolic extracts of some medicinal plants and investigate their detoxification potentials against AFB1. Phytochemical screening together with total phenolic content (TPC), total flavonoid content (TFC), and antioxidant (2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS+)) assays) were performed on nine methanolic plant extracts. Extracts were incubated with AFB1 for 24 and 48 h and liquid chromatography mass spectrometry (LC-MS) analysis done to assess their AFB1 detoxification activities. The TPC of the extracts ranged from 88.92 ± 6.54 to 210.19 ± 7.90 mg GAE/g, while TFC ranged between 4.01 ± 0.94 and 32.48 ± 1.02 mg QE/g. Radical scavenging activities of extracts varied from 4.18 ± 1.37 to 251.53 ± 9.30 µg/mL and 8.36 ± 1.65 to 279.22 ± 8.33 µg/mL based on DPPH and ABTS+ assays, respectively. Six of the plant extracts showed a time-dependent detoxification activity against AFB1 after 48 h ranging from 20.17 to 38.13 %. C. dentata bark extract showed the highest percentage of AFB1 reduction, with mean percentages of 43.57 and 70.96 % at 24 and 48 h, respectively. This was followed by C. asiatica leaves and A. melegueta seeds with a maximum of 40.81 and 38.13 %, respectively after 48 h. These extracts also possessed high TPC, TFC, and antioxidant activities compared to all the other extracts. Findings from this study demonstrate the abundance of bioactive compounds with antioxidant activity playing a role in potent AFB1 detoxification activity.

Incidence and health risk assessment of hydrogen cyanide and multi-mycotoxins in Nigerian garri.

Garri is a granular, starchy food prepared by the fermentation of mashed cassava. Hydrogen cyanide (HCN) and mycotoxins are contaminants in certain foods at different points along the food value chain. The incidence and contamination levels of HCN and multi-mycotoxins in garri from five agroecological zones of Nigeria were determined using a spectrophotometric method and ultra-high-performance liquid chromatography-tandem mass spectroscopy (UHPLC-MS/MS), respectively. The health risk associated with the consumption of contaminated garri was assessed. The health risk assessment model was used to calculate the dietary exposure of humans to the mycotoxins in garri. This was done by estimating the daily intake (EDI), the percentage tolerable daily intake (%TDI), the annual hepatocellular carcinoma (HCC) cases attributable to exposure to aflatoxins (AFs) in garri, as well as the HCC risk. The average intake of garri was estimated at 0.303 kg/day for a Nigerian adult. The incidence of HCN was 98.3% (0.056-2.463 mg/kg), and fermentation reduced the HCN level in garri more than other processing steps. The twenty-one mycotoxins identified and quantified were all within maximum levels, as applicable to those that are regulated by the EU. The %TDI for the other mycotoxins, with the exception of AFs, showed no alarming health risk with garri consumption. Annual HCC cases resulting from AF in garri were estimated at 10-60 cases for HBsAg + ve individuals and 4-23 cases for HBsAg - ve individuals based on 8.1% hepatitis B virus (HBV) incidence. Results further revealed no interdependence between HCN levels and mycotoxin content. This work suggests an unlikely chance of acute toxicity from HCN and major mycotoxins from a garri-based diet in Nigeria. Hence, it is recommended that concerned regulatory bodies maintain the existing permissible limits for HCN in Garri.

Influence of traditional dehulling on mycotoxin reduction and GC-HRTOF-MS metabolites profile of fermented maize products.

Contamination with mycotoxins has been a worldwide food safety concern for several decades, and food processing has been suggested as a potential method to mitigate their presence. In this study, the influence of traditional dehulling (TD) on the mycotoxin reduction and metabolites profile of fermented white maize products obtained via natural and three controlled fermentation methods (involving Lactobacillus fermentum, Lactobacillus plantarum, and their mixed cultures) was examined. Gas chromatography coupled with high resolution time-of-flight mass spectrometry (GC-HRTOF-MS) and ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were employed. TD brought the levels of fumonisin B1 (FB1) and B2 (FB2) in the white maize below the regulatory limit set by the European Union (EU) for maize consumed by humans. While TD increased the concentration of several mycotoxins in the fermented maize products obtained from other studied fermentation methods, it primarily reduced aflatoxin B1 (AFB1), FB1, deoxynivalenol, and 15-acetyldeoxynivalenol in the L. plantarum-fermented products. By tempering the dehulled maize, a solid-state fermentation process began. This was used in TD to make it easier to remove the pericarp. GC-HR-TOF-MS metabolomics revealed that TD brought about the generation of 12 additional compounds in the dehulled maize though some metabolites in the whole maize were lost/biotransformed. The fermented dehulled maize products obtained from the four studied fermentation procedures contained fewer compounds than the fermented whole maize products. Overall, the analysis showed that all fermented maize (whole and dehulled) produced had varied nutritional metabolites and mycotoxin concentrations below the EU maximum level, except for fermented maize obtained from mixed strains (AFB1 + AFB2 > 4.0 g/kg).

Application of a generative adversarial network for multi-featured fermentation data synthesis and artificial neural network (ANN) modeling of bitter gourd-grape beverage production.

Artificial neural networks (ANNs) have in recent times found increasing application in predictive modelling of various food processing operations including fermentation, as they have the ability to learn nonlinear complex relationships in high dimensional datasets, which might otherwise be outside the scope of conventional regression models. Nonetheless, a major limiting factor of ANNs is that they require quite a large amount of training data for better performance. Obtaining such an amount of data from biological processes is usually difficult for many reasons. To resolve this problem, methods are proposed to inflate existing data by artificially synthesizing additional valid data samples. In this paper, we present a generative adversarial network (GAN) able to synthesize an infinite amount of realistic multi-dimensional regression data from limited experimental data (n = 20). Rigorous testing showed that the synthesized data (n = 200) significantly conserved the variances and distribution patterns of the real data. Further, the synthetic data was used to generalize a deep neural network. The model trained on the artificial data showed a lower loss (2.029 ± 0.124) and converged to a solution faster than its counterpart trained on real data (2.1614 ± 0.117).

Seasonal and Geographical Impact on the Mycotoxigenicity of Aspergillus and Fusarium Species Isolated from Smallholder Dairy Cattle Feeds and Feedstuffs in Free State and Limpopo Provinces of South Africa.

This study evaluated the impact of seasonal and geographical variations on the toxigenicity of Aspergillus and Fusarium strains previously isolated from smallholder dairy cattle feeds and feedstuffs sampled during summer and winter in the Free State and Limpopo provinces of South Africa (SA). In total, 112 potential toxigenic fungal species were obtained and determined for their capability to produce mycotoxins on solid Czapek Yeast Extract Agar (CYA); followed by liquid chromatography-mass spectrometry (LC-MS/MS) analysis. Our result revealed that 41.96% of the fungal species produced their respective mycotoxins, including aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), and zearalenone (ZEN), with higher levels of AFB1 (0.22 to 1045.80 µg/kg) and AFB2 (0.11 to 3.44 µg/kg) produced by fungal species isolated from summer samples than those in winter [(0.69 to 14.44 µg/kg) and (0.21 to 2.26 µg/kg), respectively]. The same pattern was also observed for AFB1 and AFB2 in Limpopo (0.43 to 1045.80 µg/kg and 0.13 to 3.44 µg/kg) and Free State (0.22 to 576.14 µg/kg and 0.11 to 2.82 µg/kg), respectively. More so, ZEN concentrations in summer (7.75 to 97.18 µg/kg) were higher than in winter (5.20 to 15.90 µg/kg). A similar observation was also noted for ZEN in Limpopo (7.80 to 97.18 µg/kg) and Free State (5.20 to 15.90 µg/kg). These findings were confirmed via Welch and Brown-Forsythe tests with significantly (p ≤ 0.05) higher mycotoxin levels produced by fungal strains obtained in samples during summer than those in winter. In contrast, the concentrations of mycotoxins produced by the fungal species from both provinces were not significantly (p > 0.05) different.

Structure, in vitro starch digestibility and physicochemical properties of starch isolated from germinated Bambara groundnut.

This study investigated the effect of germination time (0, 24, 48 and 72 h) on the physicochemical characteristics, in vitro starch digestibility and microstructural changes in Bambara groundnut starch. The starch yield, lightness (L*) value, amylose content and resistant starch contents of isolated starches decreased significantly (p ≤ 0.05) with increasing germination time. Scanning electron microscopy revealed that starch from raw and germinated Bambara grains were smooth with no evidence of starch degradation and were mainly oval shaped, with some granule's irregular and kidney shaped. Water absorption capacity (1.33-1.90 g/g), swelling power (2.12-16.53 g/g), solubility index (1.14-13.04 g/g), and dispersibility (75.92-86.47%) greatly increased as germination timed increased. Germination did not alter the X-ray diffraction pattern (Type-A) but increased the relative crystallinity of the starches. The peak gelatinization temperatures (73.23-73.91 °C) of starch from germinated Bambara were significantly higher than native starch (72.81 °C). Native starch and starch from germinated Bambara grains had substantially high proportion of resistant starch (approx. 73%) and high pasting temperatures (approx. 88 °C). Conclusively, germination significantly changed starch structure at molecular level and impacted functionality.

Seasonal Diversity and Occurrence of Filamentous Fungi in Smallholder Dairy Cattle Feeds and Feedstuffs in South Africa.

This study investigated 65 (35 in summer and 30 in winter) smallholder dairy cattle feeds from Free State and Limpopo provinces in South Africa from 2018 to 2019 for fungal contamination and assessed the impacts of seasonal variation on fungal contamination levels, isolation frequency, and diversity. Samples were examined for fungal contamination using macro- and microscopic approaches, and their identities were confirmed by molecular means. A total of 217 fungal isolates from 14 genera, including Aspergillus, Fusarium, and Penicillium, were recovered from feeds from both seasons. The most prevalent fungal species recovered were A. fumigatus and P. crustosum. Mycological analyses showed that 97% of samples were contaminated with one or more fungal isolates, with the summer fungal mean level (6.1 × 103 to 3.0 × 106 CFU/g) higher than that of feeds sampled during winter (mean level: 1.1 × 103 to 4.1 × 105 CFU/g). Independent sample t-test revealed that the isolation frequencies of the genera Aspergillus and Fusarium were significantly (p ≤ 0.05) higher in summer than winter, while Penicillium prevalence in both seasons was not statistically (p > 0.05) different. Furthermore, the Shannon−Weiner diversity index (H') revealed a higher fungal diversity in summer (H' = 2.8) than in winter (H' = 2.1). This study on fungal contamination could be used for future fungal control and mycotoxin risk management in South Africa.

Variability in metabolites produced by Talaromyces pinophilus SPJ22 cultured on different substrates.

BACKGROUND: Several metabolites released by fungal species are an essential source of biologically active natural substances. Gas chromatography high resolution time-of-flight mass spectrometry (GC-HRTOF-MS) is one of the techniques used in profiling the metabolites produced by microorganisms, including Talaromyces pinophilus. However, there is limited information regarding differential substrates' impacts on this fungal strain's metabolite profiling. This study examined the metabolite profile of T. pinophilus strain SPJ22 cultured on three different media, including solid czapek yeast extract agar (CYA), malt extract agar (MEA) and potato dextrose agar (PDA) using GC-HRTOF-MS. The mycelia including the media were plugged and dissolved in 5 different organic solvents with varying polarities viz.: acetonitrile, dichloromethane, hexane, 80% methanol and water, and extracts analysed on GC-HRTOF-MS. RESULTS: The study revealed the presence of different classes of metabolites, such as fatty acids (2.13%), amides (4.26%), alkanes (34.04%), furan (2.13%), ketones (4.26%), alcohols (14.89%), aromatic compounds (6.38%), and other miscellaneous compounds (17.02%). Significant metabolites such as acetic acid, 9-octadecenamide, undecanoic acid methyl ester, hydrazine, hexadecane, nonadecane, eicosane, and other compounds reported in this study have been widely documented to have plant growth promoting, antimicrobial, anti-inflammatory, antioxidant, and biofuel properties. Furthermore, T. pinophilus grown on PDA and MEA produced more than twice as many compounds as that grown on CYA. CONCLUSION: Thus, our result showed that the production of essential metabolites from T. pinophilus is substrate dependent, with many of these metabolites known to have beneficial characteristics, and as such, this organism can be utilised as a sustainable and natural source for these useful organic molecules.

Mycotoxin levels and characterization of natural anti-fungal phytochemicals in pearl millet (Pennisetum glaucum) from Nigeria's six agroecological zones.

This study reports levels of multiple mycotoxins across Nigeria's six agro-ecological zones and corresponding levels of natural anti-fungal phytochemicals present in pearl millet (PM). 220 representative composite samples of PM were collected for mycotoxin analysis using ultrahigh performance liquid chromatography-mass spectrometry (UHPLC-MS), and 24 were randomly selected for determination of metabolites using gas chromatography-high resolution time of flight-mass spectrometry (GC-HRTOF-MS). In total, 15 mycotoxins were detected, all with levels below the European Union (EU) permissible limits and level of aflatoxins only up to 1.34 µg/kg. This is in sharp contrast to high levels of mycotoxins reported in maize samples from the same agroecological zones. Phytochemical analysis of the same samples identified a total of 88 metabolites, 30 of which are known anti-fungal properties from other previously published studies. The most common of these include methyl ester, bis (2-ethylhexyl) phthalate, and ç-tocopherol. The number of anti-fungal metabolites recovered from each sample ranged from 3 to 17 and varied widely in both number and composition across the agroecological zones. The anti-fungal metabolites may probably make PM less susceptible to fungal proliferation compared to other grains. Hence, it is worth exploring for possible sources of biological control products from PM.

Kinetics of Phenolic Compounds Modification during Maize Flour Fermentation.

This study aimed to investigate the kinetics of phenolic compound modification during the fermentation of maize flour at different times. Maize was spontaneously fermented into sourdough at varying times (24, 48, 72, 96, and 120 h) and, at each point, the pH, titratable acidity (TTA), total soluble solids (TSS), phenolic compounds (flavonoids such as apigenin, kaempferol, luteolin, quercetin, and taxifolin) and phenolic acids (caffeic, gallic, ferulic, p-coumaric, sinapic, and vanillic acids) were investigated. Three kinetic models (zero-, first-, and second-order equations) were used to determine the kinetics of phenolic modification during the fermentation. Results obtained showed that fermentation significantly reduced pH, with a corresponding increase in TTA and TSS. All the investigated flavonoids were significantly reduced after fermentation, while phenolic acids gradually increased during fermentation. Among the kinetic models adopted, first-order (R2 = 0.45-0.96) and zero-order (R2 = 0.20-0.82) equations best described the time-dependent modifications of free and bound flavonoids, respectively. On the other hand, first-order (R2 = 0.46-0.69) and second-order (R2 = 0.005-0.28) equations were best suited to explain the degradation of bound and free phenolic acids, respectively. This study shows that the modification of phenolic compounds during fermentation is compound-specific and that their rates of change may be largely dependent on their forms of existence in the fermented products.

Risk assessment of heavy metals in rooibos (Aspalathus linearis) tea consumed in South Africa.

A total of 80 rooibos tea samples from a range of brands were purchased from various registered retail shops in South Africa. The samples were bought during 2019 winter (40) and summer (40) period which are classified as 6 natural rooibos, 18 herbal rooibos samples, and 16 flavor rooibos samples and subjected for heavy metal analysis such as chromium (Cr), iron (Fe), arsenic (As), cadmium (Cd), and lead (Pb) using inductively coupled plasma mass spectrometer (ICP-MS). Human health risks were determined by estimating the daily intake non-cancer hazard quotient (THQ) and hazard index (HI) via oral exposure to toxic elements based on daily tea consumption. The concentration range of the determined heavy metals in rooibos tea samples were as follows: Cr (0.17-11.98 mg/kg), Fe (31-450 mg/kg), As (ND-0.51 mg/kg), Cd (0.09-0.17 mg/kg), and Pb (0.06-2.73 mg/kg). Cr was found in higher amount when compared to the World Health Organization (WHO) permissive limit (1.3 mg/kg). The concentrations of all studied heavy metals during winter and summer period were compared using two-way Anova, and no significant differences (p = 0.832) were observed for the two seasons. Both the target risk quotient (THQ) and the hazard index (HI) levels in all analyzed tea were well below 1, implying that intake of rooibos tea with analyzed heavy metals should not cause a threat to human health. On the other hand, the continuous intake due to the high concentrations of trace metals such as Cr may pose a serious chronic health risk due to accumulation in body tissues over time. The study, therefore, suggests constant monitoring of these heavy metals in teas in order to limit the risk of exceeding the permissive limits.

GC-HRTOF-MS dataset of metabolites extracted from sorghum and ting (a fermented product) produced using two strains of Lactobacillus fermentum (singly and in combination).

This data article reports the untargeted metabolite profile of whole grain sorghum (Sorghum bicolor L.) and fermented ting samples obtained using two strains of Lactobacillus fermentum. The sorghum grains were obtained from Agricol Johannesburg (South Africa) and fermentation was done at 34 °C for 24 h. Controlled fermentation with two Lactobacillus fermentum strains (L. fermentum FUA 3165 and L. fermentum FUA 3321), was done using the strains singly and in combination. The samples obtained thereafter were freeze-dried and acetonitrile/methanol/water (v/v/v) were used as extraction solvent, before analyses on a gas chromatography high resolution time of flight mass spectrometry (GC-HRTOF-MS) system. Data obtained showed the presence of different compounds, classified into metabolite groups such as acids, alcohols, benzenes, furan, esters, hydrocarbons, terpenes, phytosterols, etc., with their retention time, molecular formula, observed mass and average peak areas reported herein. These data can be used for finding biomarkers for sorghum and their derived fermented products.

Metabolite profile of Bambara groundnut (Vigna subterranea) and dawadawa (an African fermented condiment) investigation using gas chromatography high resolution time-of-flight mass spectrometry (GC-HRTOF-MS).

Metabolite profile provides an overview and avenue for the detection of a vast number of metabolites in food sample at a particular time. Gas chromatography high resolution time-of-flight mass spectrometry (GC-HRTOF-MS) is one of such techniques that can be utilized for profiling known and unknown compounds in a food sample. In this study, the metabolite profiles of Bambara groundnut and dawadawa (unhulled and dehulled) were investigated using GC-HRTOF-MS. The presence of varying groups of metabolites, including aldehydes, sterols, ketones, alcohols, nitrogen-containing compounds, furans, pyridines, acids, vitamins, fatty acids, sulphur-related compounds, esters, terpenes and terpenoids were reported. Bambara groundnut fermented into derived dawadawa products induced either an increase or decrease as well as the formation of some metabolites. The major compounds (with their peak area percentages) identified in Bambara groundnut were furfuryl ether (9.31%), bis (2-(dimethylamino)ethyl) ether (7.95%), 2-monopalmitin (7.88%), hexadecanoic acid, methyl ester (6.98%), 9,12-octadecadienoic acid (Z,Z) and 2-hydroxy-1-(hydroxymethyl)ethyl ester (5.82%). For dehulled dawadawa, the significant compounds were palmitic acid, ethyl ester (17.7%), lauric acid, ethyl ester (10.2%), carbonic acid, 2-dimethylaminoethyl 2-methoxyethyl ester (7.3%), 9,12-octadecadienoic acid (Z,Z)-, 2-hydroxy-1-(hydroxymethyl)ethyl ester (5.13%) and maltol (4%), while for undehulled dawadawa, it was indoline, 2-(hydroxydiphenylmethyl) (26.1%), benzoic acid, 4-amino-4-hydroximino-2,2,6,6-tetramethyl-1-piperidinyl ester (8.2%), 2-undecen-4-ol (4.7%), 2-methylbutyl propanoate (4.7%) and ë-tocopherol (4.3%). These observed metabolites reported herein provides an overview of the metabolites in these investigated foods, some of which could be related to nutrition, bioactivity as well as sensory properties. It is important to emphasize that based on some of the metabolites detected, it could be suggested that Bambara groundnut and derived dawadawa might serve as functional foods that are beneficial to health.

Feed-Based Multi-Mycotoxin Occurrence in Smallholder Dairy Farming Systems of South Africa: The Case of Limpopo and Free State.

Mycotoxin contamination of feed does not only cut across food and feed value chains but compromises animal productivity and health, affecting farmers, traders and consumers alike. To aid in the development of a sustainable strategy for mycotoxin control in animal-based food production systems, this study focused on smallholder farming systems where 77 dairy cattle feed samples were collected from 28 smallholder dairy establishments in the Limpopo and Free State provinces of South Africa between 2018 and 2019. Samples were analyzed using a confirmatory UHPLC-MS/MS (Ultra-high performance liquid chromatography-tandem mass spectrometry) method validated for simultaneous detection of 23 mycotoxins in feeds. Overall, mycotoxins assessed were detected across samples with 86% of samples containing at least one mycotoxin above respective decision limits; up to 66% of samples were found to be contaminated with at least three mycotoxins. Findings demonstrated that deoxynivalenol, sterigmatocystin, alternariol and enniatin B were the most common mycotoxins, while low to marginal detection rates were observed for all other mycotoxins with none of the samples containing fusarenon-X, HT-2-toxin and neosolaniol. Isolated cases of deoxynivalenol (maximum: 2385 µg/kg), aflatoxins (AFB1 (maximum: 30.2 µg/kg)/AFG1 (maximum: 23.1 µg/kg)), and zearalenone (maximum: 1793 µg/kg) in excess of local and European regulatory limits were found. Kruskal-Wallis testing for pairwise comparisons showed commercial feed had significantly higher contamination for deoxynivalenol and its acylated derivatives, ochratoxin A and fumonisins (FB1 and FB2), whereas forages had significantly higher alternariol; in addition to significantly higher fumonisin B1 contamination for Limpopo coupled with significantly higher enniatin B and sterigmatocystin for Free State. Statistically significant Spearman correlations (p < 0.01) were also apparent for ratios for deoxynivalenol/fumonisin B1 (rs = 0.587) and zearalenone/alternariol methylether (rs = 0.544).

Systematic Delineation of Media Polarity on COVID-19 Vaccines in Africa: Computational Linguistic Modeling Study.

BACKGROUND: The global onset of COVID-19 has resulted in substantial public health and socioeconomic impacts. An immediate medical breakthrough is needed. However, parallel to the emergence of the COVID-19 pandemic is the proliferation of information regarding the pandemic, which, if uncontrolled, cannot only mislead the public but also hinder the concerted efforts of relevant stakeholders in mitigating the effect of this pandemic. It is known that media communications can affect public perception and attitude toward medical treatment, vaccination, or subject matter, particularly when the population has limited knowledge on the subject. OBJECTIVE: This study attempts to systematically scrutinize media communications (Google News headlines or snippets and Twitter posts) to understand the prevailing sentiments regarding COVID-19 vaccines in Africa. METHODS: A total of 637 Twitter posts and 569 Google News headlines or descriptions, retrieved between February 2 and May 5, 2020, were analyzed using three standard computational linguistics models (ie, TextBlob, Valence Aware Dictionary and Sentiment Reasoner, and Word2Vec combined with a bidirectional long short-term memory neural network). RESULTS: Our findings revealed that, contrary to general perceptions, Google News headlines or snippets and Twitter posts within the stated period were generally passive or positive toward COVID-19 vaccines in Africa. It was possible to understand these patterns in light of increasingly sustained efforts by various media and health actors in ensuring the availability of factual information about the pandemic. CONCLUSIONS: This type of analysis could contribute to understanding predominant polarities and associated potential attitudinal inclinations. Such knowledge could be critical in informing relevant public health and media engagement policies.

Effect of Selected Cooking Ingredients for Nixtamalization on the Reduction of Fusarium Mycotoxins in Maize and Sorghum.

Although previous studies have reported the use of nixtamalization for mycotoxins reduction in maize, the efficacy of calcium hydroxide and other nixtamalization cooking ingredients for mycotoxin reduction/decontamination in sorghum and other cereals still need to be determined. The current study investigated the effect of five nixtamalization cooking ingredients (wood ashes, calcium hydroxide, sodium hydroxide, potassium hydroxide, and calcium chloride) on the reduction of Fusarium mycotoxins in artificially contaminated maize and sorghum using liquid chromatography-tandem mass spectrometry. All tested cooking ingredients effectively reduced levels of mycotoxins in the contaminated samples with reduction initiated immediately after the washing step. Except for the calcium chloride nixtamal, levels of fumonisin B1, B2, and B3 in the processed sorghum nixtamal samples were below the limit of detection. Meanwhile, the lowest pH values were obtained from the maize (4.84; 4.99), as well as sorghum (4.83; 4.81) nejayote and nixtamal samples obtained via calcium chloride treatment. Overall, the results revealed that the tested cooking ingredients were effective in reducing the target mycotoxins. In addition, it pointed out the potential of calcium chloride, though with reduced effectiveness, as a possible greener alternative cooking ingredient (ecological nixtamalization) when there are environmental concerns caused by alkaline nejayote.

Quantification of multi-mycotoxin in cereals (maize, maize porridge, sorghum and wheat) from Limpopo province of South Africa.

Mycotoxins are secondary metabolites that are produced by filamentous mycotoxigenic fungi belonging to the Alternaria, Aspergillus, Fusarium and Penicillium genera amongst others. Multi-class mycotoxins were extracted from 55 cereal samples and analysed using liquid chromatography-tandem mass spectrometry. The adopted extraction method for maize, maize porridge, sorghum and wheat was comprehensively validated. This method was further tested to determine the natural occurrence of mycotoxins in foodstuffs. Twelve (12) out of 22 mycotoxins were detected in maize, maize porridge, sorghum and wheat, including α-zearalenol (α-ZEL) (89%), fumonisin B3 (FB3) (84%), fumonisin B1 (FB1) (80%), tenuazonic acid (TeA) (78%), ochratoxin B (42%), deoxynivalenol (DON) (12%), ochratoxin A (11%), 3-acetyldeoxynivalenol (7%), sterigmatocystin (STG) (6%), 15-acetyldeoxynivalenol (2%), cyclopiazonic acid (CPA) (2%) and aflatoxin B2 (2%). The data revealed high incidence rate of α-ZEL (range: 6.5-70.5 µg kg-1) in all matrices. Maize samples had high mycotoxin co-occurrence compared to other matrices. All recovered mycotoxins in food commodities were within the maximum regulatory limits, with the exception of fumonisins (FB1 and FB3) exceeded the South African and European Commission regulation, and the highest concentration was 2153 µg kg-1 in maize. It is essential to monitor the level of emerging mycotoxins in food commodities from rural areas as trace amount of CPA (< limit of quantification), STG (range: 0.30-0.74 µg kg-1) were detected and high concentration of TeA (292.7 µg kg-1) was detected in sorghum. The occurrence of these mycotoxins further encourages frequent analyses, their co-occurrence in the samples poses a significant threat to public health and more emphasis should thus be placed on reducing the contamination levels of these toxins in staples.

Parallel validation of a green-solvent extraction method and quantitative estimation of multi-mycotoxins in staple cereals using LC-MS/MS.

In this study, 15 different mycotoxins were estimated in three staple cereals from selected agro-ecological regions in Nigeria using a 'novel' green extraction method, pressurized hot water extraction (PHWE) in comparison to a conventional solvent extraction method. Discrimination of the results of PHWE and solvent extraction using principal component analysis (PCA) and orthogonal projection to latent structures discriminate analysis (OPLS-DA) did not yield any differential clustering patterns. All maize samples (n = 16), 32% (n = 38) of sorghum and 35% (n = 37) of millet samples were positive for at least one of the 15 tested mycotoxins. Contamination levels for the cereals were higher in the warm humid rain forest region and gradually decreased towards the hot and arid region in the north of the country. The results demonstrate the applicability of PHWE as a possible alternative extraction method to conventional methods of extraction, which are solvent based.

Development, chemometric-assisted optimization and in-house validation of a modified pressurized hot water extraction methodology for multi-mycotoxins in maize.

Effective management of mycotoxins rely on stringent regulation and routine surveillance of food/feed commodities via efficient analysis, hence the continuous need for improved methods. The present study developed, optimized and validated a modified pressurized hot water extraction (PHWE) method for the simultaneous extraction of multi-mycotoxins from maize and subsequent quantification on LC-MS/MS. The PHWE system was modified using ethanol (EtOH) as a cosolvent, while a numerical modelling approach, the central composite design (CCD), was adopted for the optimization of the extraction conditions. Using the optimized method, it was possible to effectively extract and quantify 15 different mycotoxins from maize in a single step with satisfactory linearities (0.986-0.999), recoveries (14-124%) and other associated method validation parameters. Further efficacious application of the method to real samples re-affirmed the prospects of PHWE as a suitable, cost-effective and greener alternative to traditional methods of mycotoxin extraction.

Benzimidazole Schiff base derivatives: synthesis, characterization and antimicrobial activity.

A series of Schiff bases (3.a-f) bearing benzimidazole moiety was successfully synthesized in ethanol by refluxing Oct-2-ynoic acid (1,3-dihydrobenzimidazole-2-ylidene)amide with substituted amines. Fourier transform infrared (FTIR), ultra violet light (UV-VIS), elemental analysis, proton (1H) and carbon (13C) nuclear magnetic resonance spectroscopy were used to characterize the newly synthesized Schiff bases. Micro dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the Schiff bases, against 14 human pathogenic bacteria (8 Gram negative and 6 Gram positive) and against 7 fungal strains (5 Aspergillus and 2 Fusarium) representatives. Antimalarial activity against Plasmodium falciparum and antitrypanosomal property against Trypanosoma brucei was studied in vitro at a single dose concentration of the Schiff bases. Cytotoxicity of the Schiff bases was assessed against human cervix adenocarcinoma (HeLa) cells. Results obtained show that the newly synthesized Schiff bases are very potent antimicrobial agents. Gram negative bacteria Klebsiella pneumonia and Escherichia coli were more affected on exposure to Compounds 3.c-f (MIC 7.8 µg/mL) which in turn exhibited more antibacterial potency than nalidixic acid reference drug that displayed MICs between 64 and 512 µg/mL against K. pneumonia and E. coli respectively. The test compounds also demonstrated high cytotoxic effect against Aspergillus flavus and Aspergillus carbonarius as they displayed MFC 7.8 and 15.6 µg/mL. Compound 3.c exhibited the highest fungicidal property from this series with MFC alternating between 7.8 and 15.6 µg/mL against the investigated strains. The malarial activity revealed Compounds 3.c and 3.d as the more potent antiplasmodial compounds in this group exhibiting 95% and 85% growth inhibition respectively. The IC50 of Compounds 3.c and 3.d were determined and found to be IC50 26.96 and 28.31 µg/mL respectively. Compound 3.a was the most cytotoxic agent against HeLa cells in this group with 48% cell growth inhibition. Compounds 3.c, 3.d and 3.f were biocompatible with HeLa cells and displayed low toxicity. With a very low cytotoxic effect against HeLa, compound 3.c stands out to be a very good antiparasitic agent and consideration to further evaluate the candidate drug against others cell lines is necessary.