Mycobiota and mycotoxin contamination of maize flours and popcorn kernels for human consumption commercialized in Spain

Mycobiota and co-occurrence of aflatoxins, citrinin, ochratoxin A and zearalenone in 30 samples of maize flours and 30 of popcorn kernels purchased in Spain for human consumption were determined. The mycotoxin-producing ability of Aspergillus, Fusarium and Penicillium spp. was also studied.     

THE NUTRITIVE CONTENT OF AFRICAN BEERS BREWED WITH MAIZE GRITS OR SORGHUM ADJUNCT

Sorghum beer samples were brewed in a pilot plant operation using either sorghum grain or maize grits as starchy adjunct in order that the nutritive content of the beers could be compared. Significantly higher ethanol, thiamin, nicotinic acid, ash, phytate, zinc, copper, manganese, iron, magnesium, potassium and phosphorus concentrations were found in beers brewed with sorghum adjunct. Sorghum adjunct beers can therefore make a greater contribution to the nutritional requirements of the Southern African beer drinker than beers brewed with maize grits. The results obtained suggest that the use of sorghum adjunct in the brewing of sorghum beer should be encouraged for economic and dietary reasons.     

FLAVOUR AND HAZE STABILITY DIFFERENCES DUE TO HOP TANNINS IN ALL-MALT PILSNER BEERS BREWED WITH PROANTHOCYANIDIN-FREE MALT

All-malt Pilsner beers were brewed with proanthocyanidin-free malt (ant 13 × 13 × Rupal). Hopping was with n-hexane tannin-free hop extract with or without n-hexane extracted hop residue or with whole leaf hops. The different beers were analysed chemically and presented to an expert panel to detect possible preferences and differences in bitterness and astringency. The impact of hop proantho-cyanidins on haze formation is comparable to that of malt proanthocyanidins. No differences between the beers were found in triangular tests. Statistical analysis of the paired comparison tests showed that hop proanthocyanidins do not contribute to the bitterness of beer since beer brewed with tannin-free hop extract is slightly more bitter than beer brewed with whole leaf hops. No differences in astringency were noted between beers brewed with or without hop proanthocyanidins and it was found that the panel expressed a very slight preference for beer brewed with n-hexane hop extract over beer brewed with both n-hexane hop extract and extracted hop residue.     

The Microbial Contamination,Toxicity and Quality of Turned and Unturned Outdoor Floor Malted Sorghum

Turned and unturned outdoor floor malted sorghum were studied for their total microbial contamination, nature and extent of contamination by moulds, cytotoxicity (IC₅₀) and quality in terms of diastatic power (DP). The presence of aflatoxins, fumonisins, deoxynivalenol and zearalenone were also investigated. Total microbial counts were high (10⁷–10⁸ cfu/g) in both turned and unturned samples. All samples showed contamination by different moulds, with the dominant being Mucor species, Rhizopus oryzae, Fusarium moniliforme and Phoma sorghina as well as Aspergillus flavus and Alternaria alternata. The latter four are known for producing mycotoxins. Malt samples had very low cytotoxicity (IC₅₀ from 62.5 to >1000 kg/kg), though all contained fumonisins, deoxynivalenol and zearalenone at levels of <0.25–2 μg/g, 15–20 and 10–15 μg/kg, respectively. Malt DP was generally lower in turned samples compared to unturned samples probably because the heat conserved in the latter ensured better germination conditions. Overall, turning during germination did not affect the microbial load, mould population and levels of deoxynivalenol and zearalenone in sorghum malt but decreased sorghum malt DP. Thus, alternative methods of controlling the sorghum malt microbial load should be sought.     

AMINO ACID ANALYSIS OF BEER POLYPEPTIDES

The principles of amino acid analysis of proteins and polypeptides are reviewed. Analysis of the amino acid composition of dialysed beer material prepared from a wide variety of commercial and pilot brewery beers showed that the principal amino acids comprised glutamic acid/glutamine, proline, glycine and aspartic acid/asparagine. The results from the analysis of a series of pilot brewery beers brewed under standardised conditions showed that the composition of the grist may influence the amino acid composition of beer polypeptide fractions. Dialysed beer material prepared from beer brewed from grists containing torrified wheat, wheat flour and malted wheat contained greater proportions of glutamic acid/glutamine compared to material prepared from all malt beers. Further fractionation and analysis of dialysed beer material prepared from pilot brewery beers suggested that fractions MW>60000 contained polypeptide material derived from yeast mannan-protein. In addition fractions MW>60000 prepared from beer brewed from grists containing torrified wheat, wheat flour or all malted wheat may contain high molecular weight polypeptide material derived from wheat proteins. The results from the analysis of fraction MW 40,000–60000 prepared from beers brewed from grists containing all malt, 80% malt and 20% torrified wheat and 50% malt and 50% malted wheat are consistent with the presence of polypeptide material derived from cereal albumins and globulins whereas fractions MW 40,000–60000 prepared from beers brewed from 80% malt and 20% wheat flour and 100% malted wheat may contain polypeptide material derived from wheat prolamins and glutelins. The amino acid composition of fraction MW 20,000–40,000 from all pilot brewery beers investigated is consistent with the presence of polypeptide material derived from cereal prolamins and glutelins. The amino acid composition of beer polypeptide fractions may be used to detect the use of wheat adjuncts in beer brewing.     

Brewing with 100% green malt – process development and key quality indicators

Brewing with undried, germinated (green) malt has the potential to lower energy and water usage in the malting and brewing chain. However, doing so introduces technical and biochemical (flavour) challenges. Beers were brewed using 100% green malt (n = 3) or kilned pilsner malt (n = 3), prepared from the same batch in each case, utilising the pilot brewery at KU Leuven (2.5 hL). Three further pairs of beers were brewed whereby the green malt was pre-steeped under deaerated water for 1 hour; this procedure was previously shown to lower LOX activity in green malt. Six green malt beers were brewed with acceptable specifications in terms of pH, alcohol content, foam stability and colour. No significant taints or obvious defects were detected in green malt beers. Increased S-methyl methionine levels were measured in worts and beers made from green malt, however DMS concentrations in the finished beers did not differ significantly from the reference beers. Furthermore, the results demonstrated promising indicators for flavour stability, such as reduced TBI, lower residual FAN and trihydroxy fatty acid (THFA) levels in brews using untreated green malt. Using re-steep water in green malt brewing (for reasons of water economy), however, increased THFA levels, possibly because oxygen uptake was not adequately controlled at this step. Whilst further process optimisations are undoubtedly required, it is shown that an acceptable lager style beer could be brewed to a specification not dissimilar to that of a kilned malt control beer, using 100% green malt with intact rootlets. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

EXTRUDED SORGHUM AS A BREWING RAW MATERIAL

Small scale mashes (50 g total grist) with grists containing up to 50% by weight of extruded whole sorghum produced worts of high extract yield and low viscosity. Increasing the proportion of extruded sorghum in the grist resulted in decreasing wort filtration volume, total nitrogen and free amino nitrogen content. The wort filtration behaviour of mashes containing sorghum extruded at 175°C was superior to that of mashes containing sorghum extruded at 165°C or 185°C. The results from such small scale mashing experiments suggested that extruded sorghum compared favourably to extruded barley and extruded wheat as a brewing adjunct. Worts and beers were produced on a pilot brewery scale (100 1) from grists comprising 70% malt + 30% extruded sorghum and 100% malt under isothermal infusion mashing conditions. Mashes containing sorghum extruded at 175°C showed comparable wort filtration behaviour to that of the all malt control mash whereas mashes containing sorghum extruded at 165°C or 185°C showed poor wort filtration behaviour. Worts produced from grists containing extruded sorghum fermented more quickly than the control wort and attained lower values of final gravity. The resulting beers were filtered without difficulty. Beers produced from grists containing extruded sorghum contained lower levels of total nitrogen and free amino nitrogen compared to the control beer consistent with extruded sorghum contributing little or no nitrogenous material to the wort and beer. Beers brewed from grists containing extruded sorghum were of sound flavour and showed reasonable foam stability behaviour.     

Natural co-occurrence of aflatoxin and cyclopiazonic acid in peanuts grown in Argentina

Natural occurrence of aflatoxins and cyclopiazonic acid (CPA) contamination in peanuts was investigated. Co-occurrence of CPA and aflatoxins was detected in two of 50 samples analysed. The levels of these toxins found in positive samples were 4300 and 493 μg kg -1 for CPA, 625 and 435 μg kg -1 for aflatoxin B (AFB 1 ), and 625 and 83 μg kg -1 for aflatoxin G (AFG 1 ), respectively. Levels of CPA contamination in the positive samples were similar to those registered in other substrates. This is the first report of natural co-occurrence of CPA and aflatoxins in Argentina.     

Brewing Beer with Sorghum

Research reports on extracts, proteins, total nitrogen and free amino nitrogen content of sorghum malt and worts obtained from mashes indicate that sorghum is potentially an alternative substrate for conventional beer brewing in the tropics. Remarkable variations in biochemical characteristics among different sorghum cultivars affect their optimal malting conditions. Factors such as temperature and time of steeping and germinating of grains with their intrinsic enzymic activities, and kilning temperature determine the quality of malt. Further works on mashing, viscosity and fermentability of worts as well as the character of the resulting beers, such as alcoholic content, colour, taste and specific gravity tend to confirm the status of sorghum as a credible substitute for barley in beer brewing. This review reports on progress made in the use of sorghum for brewing beer.     

Application of hybrid linear ion trap-high resolution mass spectrometry to the analysis of mycotoxins in beer

This paper reports the application of liquid chromatography electrospray ionization ion trap-orbitrap mass spectrometry for the determination of 18 mycotoxins (aflatoxins, fumonisins, trichothecenes, ochratoxin A, sterogmatocystin, beauvaricin, zearalenone and zearalenol) in beer. The extraction procedure was carried out by solid phase extraction (SPE): SPE columns were conditioned with acetonitrile/methanol and water. Beer was loaded onto the column which was washed with water. In these conditions, the recoveries were more than 65% and the relative standard deviation (RSD) were below 18%. The lowest limits of quantification (LLOQ) ranged from 9 to 155?ng?ml(-1). Matrix-matched calibration was performed for each beer and reliable results were obtained from selected mycotoxins. The method was applied to the analysis of 25 commercial beers. Taking advantage of the hybrid capabilities, the presence of other mycotoxins were checked; enniatins (A, A(1), B and B(1)) and fusaproliferin were studied in all the tested samples. The survey detected the presence of zearalenone in one stout beer sample.     

100%大麦啤酒酿造过程中老化Strecker醛的研究

分别采用上面发酵工艺与下面发酵工艺进行100%大麦啤酒及100%麦芽啤酒的酿制,并对其麦汁的氨基酸含量、老化Strecker醛、自由基以及新鲜啤酒中老化Strecker醛的含量等进行了对比分析。研究发现,就麦汁而言,100%大麦麦汁中老化Strecker醛的含量都明显低于100%麦芽麦汁;同样的麦汁,上面发酵方式还原Strecker醛的能力明显优于下面发酵方式。就啤酒而言,经酵母还原后,新鲜啤酒中的老化Strecker醛含量较麦汁含量低,且100%大麦啤酒中老化Strecker醛的含量低于100%麦芽啤酒中的含量。100%麦芽麦汁的自由基含量是100%大麦麦汁的近3倍。这都预示着100%大麦啤酒的风味稳定性（新鲜度）明显好于100%麦芽啤酒。     

FLAVOUR STABILITY OF ALES BREWED USING HYDROLYSED MAIZE SYRUP—SENSORY AND ANALYTICAL ASPECTS

Whereas ales brewed entirely from malt had flavour-lives of 12 weeks, those produced from 70% malt/30% hydrolysed maize syrup had flavour-lives of at least 25 weeks. The latter beers were of sound flavour and quality, and no difficulties were experienced during their production. Their initial flavour was similar to that of beers brewed using an all-malt grist. Analysis by gas chromatography coupled mass spectrometry showed that the all-malt beer underwent a greater degree of change during natural aging than did the beer brewed with maize syrup. A number of compounds, including a series of glycerol/acetaldehyde acetals, have been identified for the first time in beer; the latter compounds were found to increase in level during aging.     

Effect of Dilute Alkaline Steeping on the Microbial Contamination,Toxicity and Diastatic Power of Sorghum Malt

Steeping in dilute alkali was investigated with the primary aim of reducing the moulds, coliforms, and preventing toxicity of sorghum malt. Red tannin‐free sorghum cultivars NK 283 and PAN 8546 were steeped in 0.1, 0.2 and 0.3% NaOH and 0.1, 0.3 and 0.5% Ca(OH)₂. The effects of these solutions on aerobic plate count, lactic acid bacteria, yeasts, moulds, coliforms, the presence of some mycotoxins (aflatoxins, fumonisins, deoxynivalenol and zearalenone), the cytotoxicity and diastatic power (DP) of sorghum malt was evaluated. Steeping NK 283 in 0.1% Ca(OH)₂ increased the DP from 10.5 to 16 Sorghum Diastatic Units (SDU)/g. However, it did not reduce the general levels of malt microflora. Steeping in dilute NaOH reduced the microflora on the malt. Steeping both grain cultivars with 0.2% NaOH resulted in malts with the moulds and coliforms reduced to approximately 3.5 and 2.0 log colony forming units/g, respectively and the percentages of some mould species to very low or undetectable levels. The DP of the 0.2% NaOH steeped malts increased to 16.2 and 26.9 SDU/g in the NK 283 and PAN 8546, respectively. There were no detectable amounts of mycotoxins and no indication of cytotoxicity in the 0.2% NaOH steeped samples. Steeping sorghum grains in food‐grade 0.2% NaOH is recommended as a method for the control of bacterial and fungal contamination during sorghum malting.     

Mycotoxin contamination of sorghum and its contribution to human dietary exposure in four sub-Saharan countries

This research aimed at evaluating the safety, and the type, level and prevalence of mycotoxins in grain sorghum of four sub-Saharan African (SSA) countries (Burkina Faso, Ethiopia, Mali and Sudan). A multi-analyte LC-MS/MS method for quantification of 23 mycotoxins (nivalenol, deoxynivalenol, fusarenon X, neosolaniol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, roquefortine C, HT-2 toxin, alternariol, T-2 toxin, FB1, FB2, FB3, zearalenone, aflatoxin G₁, aflatoxin G₂, aflatoxin B₁, aflatoxin B₂, sterigmatocystin, OTA, altenuene, alternariol monomethylether) was applied to different sorghum matrices. Of the 1533 analysed samples, 33% were contaminated with at least one of the following mycotoxins: aflatoxins, fumonisins, sterigmatocystin, Alternaria toxins, OTA and zearalenone. Country of origin, colour, source and collection period of sorghum samples significantly influenced the type, level and prevalence of mycotoxins. Sterigmatocystin (15%), fumonisins (17%) and aflatoxins (13%) were the most prevalent. FB1 (274 ± 585 µg/kg) had the highest mean concentration followed by FB2 (214 ± 308 µg/kg) while diacetoxyscirpenol (8.12 ± 19.2 µg/kg) and HT-2 (11.9 ± 0.00 µg/kg) had the lowest concentrations. Neosolaniol, fusarenon-X, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, T-2 toxin, nivalenol and roquefortine C were not detected in any of the samples. Sudan had the lowest prevalence and mean concentration of all mycotoxins. Pink sorghum had the highest concentrations of fumonisins and aflatoxins. Mycotoxins from Aspergillus spp. and Alternaria spp. are the mycotoxins of concern in SSA grain sorghum with regard to prevalence, concentration and possible health risk from exposure. Based on the performed risk characterisation, daily consumption of sorghum containing aflatoxins, alternariol, alternariol monomethyl ether, sterigmatocystin and OTA could result in exceeding the established health-based guidance values for these toxins.     

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

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

Aflatoxins,ochratoxins and zearalenone in sorghum and sorghum products in Sudan

This survey examined 60 samples of sorghum and 30 samples of sorghum products from three states (Khartoum, Kordofan and Gadarif) of Sudan for aflatoxin B1, B2, G1 and G2 (AFB1, AFB2, AFG1, AFG2), ochratoxin A and B (OTA, OTB) and zearalenone (ZEN), using high performance liquid chromatography with fluorescence detection. The limits of detection and limits of quantification were in the range 0.01–0.6 µg kg^–1 and 0.03–2.0 µg kg^?1, respectively. The frequency of contaminated samples with AFB1 from Khartoum, Gadarif and Kordofan state was 38.1%, 22.2% and 23.8%, respectively. Only two samples of sorghum from Khartoum state were contaminated with OTA (3.3%). Concentrations of OTA and OTB were low and may not cause problems. No sample of sorghum or sorghum products was contaminated with ZEN. Some sorghum samples contained AFB1 concentrations above the European Union regulatory limits. The highest contaminated samples were found in Khartoum state.     

FLAVOUR AND HAZE STABILITY DIFFERENCES DUE TO HOP AND MALT TANNINS IN ALL-MALT PILSNER BEERS BREWED WITH PROANTHOCYANIDIN-FREE AND WITH REGULAR MALT

All-malt Pilsner beers were brewed with and without hop and malt proanthocyanidins by using regular malt and whole leaf hops as well as proanthocyanidin-free malt and n-hexane tannin-free hop extract. The different beers were analysed chemically and presented to an expert panel to detect possible differences in bitterness and astringency. The impact of proanthocyanidins on the formation of haze was demonstrated clearly. No differences between the beers were found in triangular tests although their levels of total polyphenolics and anthocyanogens differed by about 225 and 85 mg/litre. Statistical analysis of the paired comparison tests showed only very slight differences between beers with or without hop and malt proanthocyanidins. Thus, proanthocyanidin-free beer was considered less than slightly more bitter and astringent than regular beer and there was a very slight preference for the latter.     

Occurrence of aflatoxins and ochratoxin A in Indian poultry feeds

From 1998 to 2001, 216 ingredients intended for incorporation into chicken feed, which included groundnut cake, maize, millets, rice bran, sorghum, soybean, sunflower, and mixed feeds, were assayed for aflatoxins and ochratoxin A contamination using an indirect competitive enzyme-linked immunosorbent assay. Thirty-eight percent of the samples were contaminated with aflatoxins and 6% with ochratoxin A. The incidence scores of aflatoxin contamination in excess of 10 microg/kg were 41 of 95 for maize, 18 of 30 for mixed feeds, 10 of 37 for groundnut, 6 of 29 for sorghum, 5 of 10 for sunflower, 3 of 14 for rice bran, and 1 of 8 for millet. Ochratoxin A contamination, in excess of 10 microg/kg, was found in 9 of 29 sorghum samples, 1 of 27 groundnut samples, 1 of 14 rice bran samples, 1 of 10 sunflower samples, and 2 of 8 millet samples. Ochratoxin A was not found in maize and mixed feeds. None of the three soybean samples contained ochratoxin A. This is the first report, to our knowledge, of co-occurrence of aflatoxins and ochratoxin A in Indian poultry feeds. The results confirm the importance of analysis of ingredients before incorporating them into mixed feeds.     

Occurrence of mycotoxigenic fungi and mycotoxins in animal feed from bihar,india

A total of 387 samples comprising animal feed, poultry feed and cattle cakes from India were examined in order to isolate mycotoxin-producing fungi as well as mycotoxins. Of 385 Aspergillus flavus group isolates 53% were capable of producing aflatoxins in liquid SMKY medium. Toxigenic strains of other mycotoxigenic fungi, viz A ochraceus, A versicolor, Penicillium citrinum and Fusarium moniliforme, were also recorded. In feed samples, beside the aflatoxins present in 139 samples zearalenone, ochratoxin A and citrinin were also recorded alone or as cocontaminants. A monsoon climate together with the socioeconomic conditions of this region are important determinants for the high incidence of mycotoxins in animal food.     

Mycobiota and co-occurrence of mycotoxins in Capsicum powder

This study aimed to: (1) determine the mycobiota of Capsicum powder samples, paying a special attention to the mycotoxigenic moulds; (2) evaluate the contamination levels of aflatoxins (AF), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), T2 and HT2 toxins in those samples. Thirty-two samples were obtained through the methods of sampling established by the European Union legislation. Aspergillus and Eurotium were the most frequently found genera. Aspergillus section Nigri had the higher relative frequency in the samples, A. niger aggregate being the most representative group of this section. Other potentially mycotoxigenic Aspergillus, Fusarium and Penicillium species were found, but in a lower frequency.Co-occurrence of mycotoxins was confirmed in the 32 Capsicum powder samples. All samples were contaminated with AF and OTA, 27% with ZEA (36% of chilli and 18% of paprika samples), 9% with DON (18% of chilli and 6% of paprika samples), 6% with T2 (18% of chilli samples) and none of the samples contained HT2.Although in the present study the most common genera found (Aspergillus and Eurotium) belong to storage moulds, some field fungi such as Fusarium spp. were also found, and their toxins were sometimes detected. This fact supports the hypothesis that mycotoxin contamination of Capsicum products may occur both in the field and/or during storage.