Phenotypic characterization of indigenous Saccharomyces cerevisiae strains associated with sorghum beer and palm wines.

In order to phenotypically characterized Saccharomyces cerevisiae strains isolated from sorghum beer and palm wines for a possible selection of a starter culture, 30 strains were tested for killer activity, temperature resistance, ethanol tolerance, carbohydrate fermentation, enzyme profile and sorghum wort fermentation. Of the tested strains, three showed a killer profile, while four showed a neutral profile and 23 were found to be sensitive to K2 toxin. Temperatures of 40 °C and 44 °C allowed to distinguish strains into four thermal groups with only three strains may grow at 44 °C. Almost tested strains were tolerant to 5% ethanol with viability rates up to 73%. But at 10% and 15% ethanol, respectively 18 and 7 strains were tolerant. Carbohydrate fermentation revealed 13 fermentation profiles, including one typical and 12 atypical profiles. The typical profile strains (16.13% of the strains) fermented glucose, galactose, fructose, sucrose, maltose, trehalose and raffinose. Most of the strains secreted lipases (mainly esterase and esterase-lipase), proteases (mainly valine and cysteine arylamidase, chrymotrypsin) and phosphatases (mainly acid phosphatase and naphthol phosphohydrolase). On contrary, only five strains isolated from sorghum beer exhibited glucosidase activity, mainly α-glucosidase. The analyse of fermented sorghum wort revealed that fermentative performance is strain dependent. Furthermore, the Hierarchical Cluster Analysis showed that the strains were separated in three distinct clusters with the strains from sorghum beer clustered separately.

Differentiation and quantification of the ochratoxin A producers Aspergillus ochraceus and Aspergillus westerdijkiae using PCR-DGGE.

Ochratoxin A (OTA) is a nephrotoxic, teratogenic, immunotoxic, and carcinogenic mycotoxin which is produced in tropical zones mainly by Aspergillus carbonarius, A. niger, A. ochraceus, and A. westerdijkiae. A. ochraceus and A. westerdijkiae species are phenotypically and genomically very close but A. westerdijkiae produce OTA at a very higher level than A. ochraceus. These species have been differentiated recently. The DNA primer pairs which were drawn so far are not specific and a genomic region of the same size is amplified for both species or they are too specific, and in this case, the DNA of a single species is amplified. To help preventing OTA contamination of foodstuffs, the PCR-DGGE (Denaturing Gradient Gel Electrophoresis) method was used to discriminate between A. ochraceus and A. westerdijkiae DNA fragments of the same size but with different sequences and thus faster access to a diagnosis of the toxigenic potential of the fungal microflora. The proposed methodology was able to differentiate A. westerdijkiae from A. ochraceus with only one primer pairs in a single run. A calibration based on initial DNA content was obtained from image analysis of the DGGE gels and a method of quantification of the two strains was proposed.

Microbiological safety of flours used in follow up for infant formulas produced in Ouagadougou, Burkina Faso.

The prevalence of diarrheal diseases in children aged from 6 to 24 months in Burkina Faso is 38%. These diarrheas may be due to the consumption of contaminated weaning food. Therefore, the microbiological quality of follow up infant flours used as supplement foods is a key-point to reduce children diseases. In this study, the microbiological safety of locally-produced infant flours was investigated. One hundred and ninety-nine (199) samples were collected mainly in retails outlets and in Recovery and Nutrition Education Centers. According to the Burkina Faso regulations, microbiological analyses were carried out for Total Aerobic Mesophilic Flora (TAMF), thermotolerant coliforms, Salmonella spp. and yeasts/molds. The bacterial and fungal isolates were identified using phenotypic and genotypic methods and the study of the production of mycotoxins was carried out from the fungal isolates. In collected samples, the TAMF count ranged from 0 to 1.8 × 106 CFU/g with a total average of 6.3 × 104 CFU/g. About 2% of the samples had a microbial load exceeding the standards (105 CFU/g). No Salmonella spp. was isolated in the final infant flours. However, the presence of Enterobacteriaceae (Klebsiella spp. Enterobacter spp. and Cronobacter spp.) was detected and molecular characterization revealed also the presence of fungal species of the genus Aspergillus spp., Penicillium spp. and Fusarium spp. Some of these species were found to produce aflatoxins, ochratoxin A and fumonisins, which are potential carcinogenic toxins. These results demonstrated the need for a preventive approach based on the application of Hazard Analysis Critical Control Point in the food industry to ensure food safety of infant flours in Burkina Faso.

Aspergillus korhogoensis, a Novel Aflatoxin Producing Species from the Côte d'Ivoire.

Several strains of a new aflatoxigenic species of Aspergillus, A. korhogoensis, were isolated in the course of a screening study involving species from section Flavi found contaminating peanuts (Arachis hypogaea) and peanut paste in the Côte d'Ivoire. Based on examination of four isolates, this new species is described using a polyphasic approach. A concatenated alignment comprised of nine genes (ITS, benA, cmdA, mcm7, amdS, rpb1, preB, ppgA, and preA) was subjected to phylogenetic analysis, and resulted in all four strains being inferred as a distinct clade. Characterization of mating type for each strain revealed A. korhogoensis as a heterothallic species, since three isolates exhibited a singular MAT1-1 locus and one isolate exhibited a singular MAT1-2 locus. Morphological and physiological characterizations were also performed based on their growth on various types of media. Their respective extrolite profiles were characterized using LC/HRMS, and showed that this new species is capable of producing B- and G-aflatoxins, aspergillic acid, cyclopiazonic acid, aflavarins, and asparasones, as well as other metabolites. Altogether, our results confirm the monophyly of A. korhogoensis, and strengthen its position in the A. flavus clade, as the sister taxon of A. parvisclerotigenus.

Study of selenium distribution in the protein fractions of the Brazil nut, Bertholletia excelsa.

The high selenium content of the Brazil nut, Bertholletia excelsa, makes this seed a healthy food qualified as an antiradical protector. The studied nut contained 126 ppm of selenium. Selenium was found to be distributed in the nut protein fractions. The water-extracted fraction, which represented 17.7% of the cake protein, was the richest in selenium with 153 ppm. Analysis by HPLC-MS showed that selenium was linked by a covalent bond to two amino acids to form selenomethionine and selenocystine. The selenomethionine represented a little less than 1% of the total amount of methionine.

Use of ATP bioluminescence to determine the bacterial sensitivity threshold to a bacteriocin.

A new ATP bioluminescence-based method was developed to determine the effectiveness of nisin on a sensitive strain of Lactococcus cremoris. The principle of the method is to quantify the release of adenylic-nucleotides (AN) by a sensitive strain under the action of the bacteriocin, with the complex luciferin-luciferase. Nisin-induced leakage of AN included ATP from a sensitive L. cremoris to the external medium immediately after the contact with the bacteria. The growth of L. cremoris was correlated with the extracellular AN content. The extracellular ATP and AN concentration exhibited a linear correlation to the logarithm of the nisin concentration. For the determination of the effectiveness threshold, the concentration of AN was more sensitive and more reliable than the direct quantification of ATP. The effectiveness threshold, corresponding to a 100% inhibition of L. cremoris growth, was obtained for a null concentration of intracellular nucleotides, i.e. for a AN(tot):AN(ext) ratio = 1. For an initial concentration of 1.4 x 10(7) bacteria/mL, the nisin effectiveness threshold is 3.4 +/- 0.01 mg nisin/L. It is possible to detect effectiveness threshold concentration by taking into account the physiological state of the cells.