Contamination with storage fungi of human food from Cameroon

In a mycological study, a total of 95 human food samples were investigated to evaluate the incidence of fungal contamination in Cameroon by conventional identification method and partly confirmed by DNA sequencing. The isolated fungal spp. were further studied to determine their toxigenic potentials. The investigation revealed the predominance of Aspergillus and Penicillium with 96% of samples contaminated with at least one species of these fungi, whereas the incidence of co-contamination of samples was 85%. Aspergillus flavus and Aspergillus parasiticus (Flavi section) were the most predominant species contaminating mainly maize and peanuts. In addition, P. crustosum and P. polonicum were the most common contaminants belonging to the genus Penicillium. On the other hand, A. ochraceus (Circumdati section) registered a low incidence rate of 5%, including other members of the Aspergillus group. Other members of the genera Rhizopus and Alternaria spp. were also registered in the study. A majority of fungal strains of A. ochraceus, A. parasiticus, P. crustosum and P. polonicum isolated were toxigenic, producing the mycotoxins tested for, while none was detected in cultures of A. fumigatus. The high incidence rate of fungi contamination coupled with their potentials in producing mycotoxins gives a strong indication that the samples tested may likely be contaminated with various mycotoxins. There is need for further study to assess the incidence of mycotoxins contamination in similar food samples.     

Mycobiota of cocoa: from farm to chocolate

The present work was carried out to study the mycobiota of cocoa beans from farm to chocolate. Four hundred and ninety-four samples were analyzed at various stages of cocoa processing: (i) primary stage at the farm (fermentation, drying, and storage), (ii) secondary stage at processing (testa, nibs, liquor, butter, cake and powder) and (iii) the final chocolate product (dark, milk, white and powdered) collected from retail outlets. Direct plating or dilution plating on Dichloran 18% Glycerol agar were used for cocoa beans and processed product analyses, respectively. Fungi were isolated and identified using different keys of identification. The largest numbers and diversity of fungi were observed in the samples collected at the farm, especially during drying and storage. The species with the highest occurrence among samples were: Absidia corymbifera, Aspergillus sp. nov., A. flavus, Penicillium paneum and yeasts. A total of 1132 potentially toxigenic fungi were isolated from the following species or species groups: A. flavus, Aspergillus parasiticus, Aspergillus nomius, Aspergillus niger group, Aspergillus carbonarius and Aspergillus ochraceus group. The highest percentage of toxigenic fungi was found at the drying and storage stages. The industrial processing reduced the fungal contamination in all fractions and no fungi were found in the final chocolate products. The knowledge of which fungi are dominant at each processing stage of cocoa provides important data about their ecology. This understanding leads to a reduction in fungal spoilage and mycotoxin production in this product.     

Mercury content of some wild edible mushrooms

 The mercury (Hg) content of 112 samples of common, wild edible mushrooms was analysed by atomic absorption spectrometry. The average Hg content of all the samples was 1.72 mg/kg dry mass (DM) but the Hg concentrations found in different taxonomic groups (genera and species varied) remarkably. Although different sampling locations obviously have an effect on the Hg concentration of sporocarps, we found evidence for a Hg-accumulating capacity of some taxonomic groups. In addition to Agaricus and Macrolepiota, where the phenomenon of mercury accumulation has already been described, our results indicate high Hg levels in samples of Lycoperdon perlatum (average 2.94 mg/kg DM) and in Lepista species (average 3.02 mg/kg DM). These data confirm the need for greater attention to be given to Hg levels in wild common mushrooms, especially to Hg-accumulating species. Received: 20 February 1997     

Mercury content of some wild edible mushrooms

 The mercury (Hg) content of 112 samples of common, wild edible mushrooms was analysed by atomic absorption spectrometry. The average Hg content of all the samples was 1.72 mg/kg dry mass (DM) but the Hg concentrations found in different taxonomic groups (genera and species varied) remarkably. Although different sampling locations obviously have an effect on the Hg concentration of sporocarps, we found evidence for a Hg-accumulating capacity of some taxonomic groups. In addition to Agaricus and Macrolepiota, where the phenomenon of mercury accumulation has already been described, our results indicate high Hg levels in samples of Lycoperdon perlatum (average 2.94 mg/kg DM) and in Lepista species (average 3.02 mg/kg DM). These data confirm the need for greater attention to be given to Hg levels in wild common mushrooms, especially to Hg-accumulating species. Received: 20 February 1997     

Diversity of lactic acid bacteria in suan-tsai and fu-tsai, traditional fermented mustard products of Taiwan

Fu-tsai and suan-tsai are spontaneously fermented mustard products traditionally prepared by the Hakka tribe of Taiwan. We chose 5 different processing stages of these products for analysis of the microbial community of lactic acid bacteria (LAB) by 16S rRNA gene sequencing. From 500 LAB isolates we identified 119 representative strains belonging to 5 genera and 18 species, including Enterococcus (1 species), Lactobacillus (11 species), Leuconostoc (3 species), Pediococcus (1 species), and Weissella (2 species). The LAB composition of mustard fermented for 3 days, known as the Mu sample, was the most diverse, with 11 different LAB species being isolated. We used sequence analysis of the 16S rRNA gene to identify the LAB strains and analysis of the dnaA, pheS, and rpoA genes to identify 13 LAB strains for which identification by 16S rRNA gene sequences was not possible. These 13 strains were found to belong to 5 validated known species: Lactobacillus farciminis, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Weissella cibaria, and Weissella paramesenteroides, and 5 possibly novel Lactobacillus species. These results revealed that there is a high level of diversity in LAB at the different stages of fermentation in the production of suan-tsai and fu-tsai.     

Selected arsenic species: As(III), As(V) and dimethylarsenic acid (DMAA) in Xerocomus badius fruiting bodies

The aim of the study was to determine the content of As(III), As(V) and DMAA (dimethylarsinic acid) in Xerocomus badius fruiting bodies collected from selected Polish forests from areas subjected to very low or high anthropopressure and some commercially available samples obtained from the Polish Sanitary Inspectorate. The arsenic species determination was provided by two independent HPLC–HG-AAS hyphenated systems. The results show high levels (up to 27.1, 40.5 and 88.3 mg kg⁻¹ for As(III), As(V) and DMAA, respectively) of arsenic and occurrence of different species in mushrooms collected from areas subjected to high anthropopressure and two commercially available samples. For mushroom samples collected from areas not subjected to high anthropopressure and two commercially available samples the arsenic species level was below 0.5 mg kg⁻¹ for each arsenic form. Therefore, the accumulation of arsenic by mushrooms may lead to high (toxic for humans) arsenic concentrations, and arsenic species levels should be monitored in mushroom foodstuffs.     

Interspecific associations among stored-grain beetles

Recent increases in prices of raw grain, including wheat, will reduce action thresholds for insect damage and therefore justify more research into management practices and understanding of pest ecology in stored grain. Compared to most other habitats, natural or man-made, a filled grain silo constitutes a unique and fairly homogeneous habitat in which food availability for many grain-feeding insects is unlimited. A fundamental aspect of stored-grain insect ecology is a better understanding of associations among common beetle species. We analyzed the densities of three important stored-grain beetle species, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) in wheat samples collected in 1999–2001 from 129 grain silos in Kansas. The beetles studied here are highly mobile, and the number of insects in each grain sample is a result of the beetles' preference for favorable micro-environmental conditions and possibly of intra- and interspecific associations. In general, the number of T. castaneum in a grain sample increased as the number of R. dominica increased, but the number of C. ferrugineus was not correlated with the number of R. dominica. The densities of both T. castaneum and R. dominica decreased as the number of C. ferrugineus increased. Cryptolestes ferrugineus and T. castaneum can be predators and the species composition of insects in a grain sample may be modified by predation. As T. castaneum populations increased, so did R. dominica but not C. ferrugineus. Our analysis of the species composition in grain samples is discussed in an ecological context.     

Colour stability of canned strawberries using black carrot and elderberry juice concentrates as natural colourants

Besides texture degradation and aroma loss colour fading and browning of processed fruits, in particular strawberries, displaying comparatively low amounts of non-acylated anthocyanins, still pose a serious problem. Therefore, in the present study the colour of texture-improved canned strawberries was stabilised using commercial concentrates from black carrots and elderberries as natural colourants. CIE L ^* a ^* b ^* values of the fruit surface and of the brine solutions were monitored over a period of 24 weeks under light exposure. Furthermore, the contents of individual anthocyanins were analysed by HPLC-DAD. The colour of the processed control fruits (without added colourant) was unacceptable even at the beginning of the storage. Their anthocyanin contents were reduced by more than 85% at the end of the storage period. In contrast, the colour of canned strawberries containing black carrot and elderberry anthocyanins was significantly improved. Colour difference values ΔE ^* of black carrot-coloured strawberries were found to be smaller than those of the samples containing elderberry concentrate as a colourant. This could be ascribed to the presence of acylated anthocyanins in the first exhibiting exceptional storage stability, while all non-acylated pigments were degraded. The chill brining process for fruit firming markedly affected pigment stability, which can be attributed to enzymes that are still active at cold storage conditions.     

DNA Arrays and Membrane Hybridization Methods for Screening of Six Lactobacillus Species Common in Food Products

Dot blot and deoxyribonucleic acid (DNA) array hybridization assays for the traceability of Lactobacillus species in food have been developed to monitor and validate typical food products. A primer set was designed to amplify the 540-bp region located at +157 of the tuf (Elongation factor Tu) gene of the Lactobacillus genus. An oligonucleotide array, containing 73 Lactobacillus species-specific tuf sequences representing 21 species, was developed and tested for identifying L. paracasei, L. rhamnosus, L. plantarum, and L. buchneri. We also tested a rapid screening method for monitoring the species present in airy samples. Dot blot hybridization identified polymerase chain reaction amplicons immobilized on nylon membranes, using six tuf-based cyanine-3-labeled 18-mer oligonucleotides, specific for L. paracasei, L. zeae, L. fermentum, L. plantarum, L. rhamnosus, and L. buchneri. This method discriminates between multiple species of Lactobacilli isolated directly from cheese samples, simultaneously. The tuf gene sequences, verified here with the DNA array method and used in dot blot hybridization, were shown to be a reliable tool for the simultaneous detection and differentiation of four Lactobacillus species. The hybridization techniques developed in this study may be useful in food processing and the analysis of food origin traceability.     

Chemical composition and antimicrobial activity of the essential oils of four Ocimum species growing in Tanzania

As part of ongoing research on Tanzanian plants used as edibles or spices, six samples of essential oils from four Ocimum species (O. basilicum, O. kilimandscharicum, O. lamiifolium, O. suave) were analyzed by GC and GC–MS. Eighty-one compounds, corresponding to 81.1–98.2% of the chemical components of the oils, were identified. Major compounds were either phenyl propane derivatives or terpenoids, including methyl eugenol, 1,8-cineole, camphor, bornyl acetate, germacrene-D, E-myroxide, germacrene-B, caryophylene oxide and p-cymene. The oils were also evaluated for antimicrobial activity against eight bacterial strains and three fungi. The oil of O. suave (B) showed the strongest antibacterial activity; O. suave (A), O. kilimandscharicum and, O. lamiifolium were moderately active, while O. basilicum oil was weakly active. However, none of the oils was active against the fungi species. The study has shown that, Ocimum oils could potentially be used as anti-infective agents.     

Bio-guided fractionation of the antimutagenic activity of methanolic extract from the fruit of Randia echinocarpa (Sessé et Mociño) against 1-nitropyrene

Randia echinocarpa is a native plant from Mexico that produces an edible fruit with several ethnopharmacological uses (e.g. cancer, kidney ailments, and diabetes). Extracts of this fruit have shown antimutagenic activity. In this report, a methanolic extract of R. echinocarpa and a bio-guided chromatographic strategy were used to obtain an hexanic fraction (HF) with strong antimutagenic activity (microsuspension assay with Salmonella typhimurium YG1024) using 1-nitropyrene as mutagen (1-NP, 50 and 100 ng/tube). The HF (500 ng/tube) showed the highest inhibition percentage of mutagenic activity (PI) (75%, 1-NP 50 ng/tube; 84%, 1-NP 100 ng/tube). HF chromatography with silica produced HF₁ which was further separated to produce the fractions with the highest antimutagenic activities (HF_1–1 and HF_1–2, PI ≥ 60%). These fractions were chemically characterized by chromatography and gas chromatography–mass spectrometry; among the main components of HF, HF_1–1 and HF_1–2 were registered linoleic acid, palmitic acid and β-sitosterol, which could be responsible for the antimutagenic activity of R. echinocarpa fruit. The samples evaluated were neither toxic nor mutagenic. Randia echinocarpa is an underutilized plant and its fruit has been used traditionally as food/medicine; fruit consumption could provide human health benefits and it has potential to be exploited under conditions of ecological sustainability.     

Production of nitric oxide (NO) by lactic acid bacteria isolated from fermented products

In this study, the bacteria which were isolated from various milk and fermented food products were tested for their ability to convert metmyoglobin to nitrosomyoglobin. Lactic acid bacteria were isolated from samples of raw milk, unsalted butter, Beyaz cheese, yoghurt, pickles and silage. The nitric oxide (NO) forming abilities of 1534 isolates were tested using plates of de Man, Rogosa, Sharpe agar supplemented with metmyoglobin (MRS-Mb). Ten isolates formed bright red colonies, brown or clear zones due to the conversion of metmyoglobin to nitrosomyoglobin were identified. Five of the 10 bacteria were identified as Lactobacillus plantarum, three as Pediococcus acidilactici, and two as Leuconostoc mesenteroides subsp. dextranicum. NO formation ability was measured in MRS-Mb broth. There were differences not only among the species, but also among the strains of a species. The highest NO concentrations of 51.5, 51.3, 50.2 μM were produced by P. acidilactici S2, L. plantarum T119, and P. acidilactici S3, respectively.     

The occurrence of Listeria monocytogenes in cheese from a manufacturer associated with a case of listeriosis

A case of listeriosis was associated with the consumption of a soft cheese produced in England. Goats cheese and other products from the same food manufacturer were examined for the presence of Listeria over the following 11 months. Listeria monocytogenes was isolated from 16 of 25 cheese samples on retail sale, 12 of 24 cheese samples obtained directly from the factory, and from shelving within the plant. Phage-typing of 68 isolates of L. monocytogenes from cheese samples and the factory showed that 66 (97%) were indistinguishable from the strain isolated from the patient's cerebrospinal fluid and stool. L. monocytogenes was not isolated from seven goats milk or two yoghurt samples. Listeria innocua was isolated from 10 cheese samples, two of which contained no other species of Listeria. Levels of L. monocytogenes shortly after production were low (<10/g), but were higher (10⁵–10⁷ cfu/g) in six of the 16 cheese samples obtained from retail outlets. Multiplication of L. monocytogenes was demonstrated in cheeses contaminated at the factory and held at 4°C in the laboratory.     

Authentication of three valuable <Emphasis Type="Italic">Dendrobium</Emphasis> species by adapter ligation-mediated allele-specific amplification

The adapter ligation-mediated allele-specific amplification (ALM-ASA) has been applied to authenticate three valuable Dendrobium species (Dendrobium aphyllum, Dendrobium devonianum and Dendrobium officinale) and their corresponding “Fengdou” foods samples. Three species-specific primers and a universal primer were designed. By using these primers, three amplification products are 198 bp (D. aphyllum), 250 bp (D. devonianum) and 310 bp (D. officinale) in length, which were clearly distinguishable on a 2.0% agarose gel. In addition, no band was observed with other 17 allied Dendrobium species of “Fengdou”. The results showed that ALM-ASA could be used to authenticate not only the three valuable Dendrobium species but related food products such as “Fengdou” foods by simultaneous detection of three-site nucleotide difference on rDNA internal transcribed spacer region.     

Sequence characterized amplified region markers: A reliable tool for adulterant detection in turmeric powder

Turmeric powder (Curcuma longa L.), an important medicinal spice product traded internationally, is subjected to adulteration by design or default with powders of related curcumin containing wild species like Curcuma zedoaria and Curcuma malabarica leading to toxicity and poor quality of the produce. The present study aims at development of specific, sensitive and reproducible Sequence Characterized Amplified Region (SCAR) markers to detect these adulterants in traded turmeric powder. Two putative RAPD markers, ‘Cur 01’ and ‘Cur 02’, generated by random primers OPA 01 and OPE 18 were identified as C. zedoaria/C. malabarica specific by comparative RAPD analysis of genuine turmeric and market samples of turmeric powder, C. zedoaria and C. malabarica. These specific RAPD markers were cloned and sequenced. Two pairs of SCAR primers were designed from the RAPD markers ‘Cur 01’ and ‘Cur 02’, respectively. Six market samples of turmeric powder and four simulated standards besides the genuine samples were analyzed using the specific SCAR markers. Both the SCAR markers detected the presence of C. zedoaria/C. malabarica adulteration in four market samples and all the simulated standards prepared in different concentrations. The two SCAR markers developed in the study would be potentially useful for the regulatory agencies to detect C. zedoaria/C. malabarica adulteration in traded turmeric powder. The analytical strategy being very simple could be used for large scale screening of turmeric powder samples intended for export and domestic uses.     

Pathogenic Yersinia enterocolitica 2/O:9 and Yersinia pseudotuberculosis 1/O:1 strains isolated from human and non-human sources in the Plateau State of Nigeria

Foodborne yersiniosis, caused by enteropathogenic Yersinia, especially Yersinia enterocolitica, is an important cause of diarrhea in developed countries, especially in temperate zones. Since studies concerning the presence of enteropathogenic Yersinia in humans and foods are rare in developing countries and tropical areas, human and non-human samples were studied in Plateau state of Nigeria to obtain information on the epidemiology of Y. enterocolitica and Yersinia pseudotuberculosis. Surprisingly, ail-positive Y. enterocolitica and inv-positive Y. pseudotuberculosis were isolated in Plateau state of Nigeria from several samples of human and non-human origin. Bioserotype 1/O:1 was the only Y. pseudotuberculosis type found. Y. enterocolitica belonging to bioserotype 2/O:9 was the dominating type found in most samples. Bioserotype 4/O:3 was isolated only from one pig and one sheep. Using PFGE, 5 genotypes were obtained among 45 Y. enterocolitica 2/O:9 strains with NotI, ApaI and XhoI enzymes and 3 among 20 Y. pseudotuberculosis 1/O:1 strains with NotI and SpeI enzymes. All human Y. pseudotuberculosis 1/O:1 strains were indistinguishable from pig, sheep or food strains. The dominating genotype of Y. enterocolitica 2/O:9 strains among humans was also found among strains isolated from pig, fermented cow milk and traditional intestine pepper soap samples.     

Assessment of the microbiological safety of salad vegetables and sauces from kebab take-away restaurants in the United Kingdom

The purpose of this study was to establish the microbiological safety of salad vegetables and sauces served in kebab take-away restaurants. Comparison with published microbiological guidelines revealed that 4.7% of 1213 salad vegetable samples were of unsatisfactory microbiological quality due to Escherichia coli and/or Staphylococcus aureus levels at ≥10² cfu g⁻¹. Another 0.3% of salad samples were of unacceptable quality due to S. aureus at ≥10⁴ cfu g⁻¹ (2 samples) or the presence of Salmonella Kentucky (1 sample). Cucumber was the most contaminated salad vegetable with regards to unsatisfactory levels of E. coli (6.0%) or S. aureus (4.5%). Five percent of 1208 sauce samples were of unsatisfactory microbiological quality due to E. coli, S. aureus at ≥10² cfu g⁻¹ and/or Bacillus cereus and other Bacillus spp. at ≥10⁴ cfu g⁻¹. A further 0.6% of sauce samples were of unacceptable quality due to Bacillus spp. (Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis) at ≥10⁵ cfu g⁻¹ or the presence of Salmonella Agbeni (1 sample). More samples of chilli sauce (8.7%) were of unsatisfactory or unacceptable microbiological quality than any other sauce types. The results emphasize the need for good hygiene practices in kebab take-away restaurants handling these types of ready-to-eat products.     

Characterization of Lactobacillus isolates from fermented olives and their bacteriocin gene profiles

Near one hundred isolates of Lactobacillus paraplantarum, Lactobacillus pentosus and Lactobacillus plantarum from table olives were studied. Strains were genotyped by rep-PCR. Although the technique failed to differentiate some isolates at the species level, it proved a robust and easy procedure that could be useful for distinguishing between related strains of L. paraplantarum, L. pentosus and L. plantarum from a large pool of unrelated strains of these species. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnB, plnC, plnD, plnE/F, plnF, plnI, plnJ, plnK, plnG and plnN in most isolates of the three species. Sequences of bacteriocin genes present in L. paraplantarum and L. pentosus were homologous to L. plantarum genes. Through a discriminating analysis of the bacteriocin gene profiles, it was possible to establish a relationship between the origin of isolation and the LAB isolates, regardless of species.     

Changes in quality and phenolic compounds of virgin olive oils during objectively described fruit maturation

The effect of fruit ripening on the quality of the oil extracted and on the changes in the amount of phenolic compounds was determined in two olive varieties (Olea europaea, cvs. Arbequina and Picual) in two crop seasons, characterized by showing the same mean temperature and different rainfalls. Maturation level was evaluated using six methods: Harvest date, ripening index (RI), fruit skin colour, fruit firmness, and amount of chlorophylls and carotenoids in the oil. Oil quality, evaluated using the parameters established to determine the quality level of virgin olive oils (acidity, K₂₃₂, K₂₇₀, peroxide index, and panel test), was not affected by fruit ripening or by the increase in rainfall of the season. However, the changes in oil stability and phenolic compounds in the oils extracted during fruit ripening strongly differed according to the variety, the maturity level of the fruit and the crop season tested. Fruit skin colour and firmness allowed a better discrimination at the initial maturity stages than the other methods tested.