Carmen Wacher

Lactic acid bacterial diversity in the traditional mexican fermented dough pozol as determined by 16S rDNA sequence analysis.

The lactic acid bacteria diversity of pozol, a Mexican fermented maize dough, was studied using a total DNA extraction and purification procedure and PCR amplification of 16S rDNA for gram-positive and related bacterial groups. Thirty-six clones were obtained and sequenced to 650 nucleotides. These partial sequences were identified by submission to the non-redundant nucleotide database of NCBI. The identified sequences were aligned with reference sequences of the closest related organisms. This analysis indicated that only 14 sequences were unique clones and these were identified as Lactococcus lactis, Streptococcus suis, Lactobacillus plantarum, Lact. casei, Lact. alimentarium, and Lact. delbruekii and Clostridium sp. Two non-ribosomal sequences were also detected. Unlike other environments analyzed with this molecular approach where many unidentified microorganisms are found, the identity of most sequences could be established as lactic acid bacteria, indicating that this is the main group among the gram-positive bacteria in pozol. Use of this molecular method permitted detection of lactic acid bacteria different from those previously isolated and identified by culture techniques

Sources of microorganisms in pozol, a traditional Mexican fermented maize dough.

Freshly prepared pozol, a traditional Mexican fermented maize dough, contained (c.f.u./g wet wt): lactic acid bacteria, 104 to 106; aerobic mesophiles, 104 to 105; Enterobacteriaceae, 102 to 103; yeasts, 102 to 104; and mould propagules, <103. After 30 h at 28°C the numbers were, respectively: 109, 7×106, 5×105, 106 and 104. Soaking alkali-treated grains overnight allowed lactic acid bacteria, aerobic mesophiles and Enterobacteriaceae to grow and these then constituted the primary microbial flora of the pozol dough. Grinding in a commercial mill inoculated the dough with lactic acid bacteria, aerobic mesophiles, Enterobacteriaceae and yeasts. Other processing stages, including the nature of the surface upon which the balls were made, handling of the dough, and air, contributed only minor numbers of microbes compared with the two major sources, soaking and grinding. The pH of pozol fell from an initial value of 7.3 to 4.6 after 30 h incubation at 28°C. The numbers of Enterobacteriaceae and other aerobic mesophilic bacteria remained constant between 11 and 30 h incubation and there was no evidence of the acidic conditions having any lethal effects on these organisms.

Survival and characterization of Escherichia coli strains in a typical Mexican acid-fermented food.

In this study, the presence and pathogenic characteristics of Escherichia coli strains in pozol, an acid-fermented maize beverage consumed in South-eastern Mexico, were determined. Seventy-three E. coli strains were isolated at early and late times (6 and 48 h) during the pozol fermentation process, when pH values of the doughs were 6.7-4.7 (6 h) and 4.7-3.7 (48 h). Serotypes that belong to diarrheagenic E. coli serogroups O18, O88, O8, O11, O20, O173 were identified. HEp-2 cell adherence in vitro assays showed localized, diffuse and aggregative adherence patterns among some of these strains. A DNA colony hybridization analysis with different probes showed the presence of virulence genes related to diarrheal pathogenesis. Thirty-three percent of the E. coli strains were tetracycline-resistant and 95% had a 20 kb plasmid. The presence and survival of potentially pathogenic E. coli in acid-fermented pozol suggest that such foods may be a potential source of foodborne outbreaks.

Chemical and Nutritional Composition of Tejate, a Traditional Maize and Cacao Beverage from the Central Valleys of Oaxaca, Mexico

Foam-topped cacao and maize beverages have a long history in Mesoamerica. Tejate is such a beverage found primarily in the Zapotec region of the Central Valleys of Oaxaca, Mexico. Historically tejate has been ceremonially important but also as an essential staple, especially during periods of hard fieldwork. However, the nutritional contribution of traditional foods such as tejate has not been investigated. We analyzed tejate samples from three Central Valley communities, vendors in urban Oaxaca markets and one migrant vendor in California, USA for their proximate composition, amino acid content and scores, and mineral and methylxanthine content. Nutritional and chemical variation exists among tejate recipes, however, the beverage is a source of energy, fat, methylxanthines, K, Fe and other minerals although their availability due to presence of phytates remains to be determined. Tejate is a source of protein comparable to an equal serving size of tortillas, with protein quality similarly limited in both. Tejate provides the nutritional benefits of maize, and some additional ones, in a form appealing during hot periods of intense work, and year round because of its cultural significance. Its substitution by sodas and other high glycemic beverages may have negative nutritional, health and cultural consequences.

Quantitative analyses of the bacterial microbiota of rearing environment, tilapia and common carp cultured in earthen ponds and inhibitory activity of its lactic acid bacteria on fish spoilage and pathogenic bacteria

The present study aimed to evaluate the bacterial load of water, Nile Tilapia and common Carp intestines from earthen ponds, isolate lactic acid bacteria (LAB) and assess their antimicrobial activity against fish spoilage and pathogenic bacteria. Following enumeration and isolation of microorganisms the antimicrobial activity of the LAB isolates was evaluated. Taxonomic identification of selected antagonistic LAB strains was assessed, followed by partial characterisation of their antimicrobial metabolites. Results showed that high counts (>4 log c.f.u ml−1 or 8 log c.f.u g−1) of total aerobic bacteria were recorded in pond waters and fish intestines. The microbiota were also found to be dominated by Salmonella spp., Vibrio spp., Staphylococcus spp. and Escherichia coli. LAB isolates (5.60%) exhibited potent direct and extracellular antimicrobial activity against the host-derived and non host-derived spoilage and pathogenic bacteria. These antagonistic isolates were identified and Lactococcus lactis subsp. lactis was found as the predominant (42.85%) specie. The strains displayed the ability to produce lactic, acetic, butyric, propionic and valeric acids. Bacteriocin-like inhibitory substances with activity against Gram-positive and Gram-negative (Vibrio spp. and Pseudomonas aeruginosa) bacteria were produced by three L. lactis subsp. lactis strains. In this study, the LAB from the microbiota of fish and pond water showed potent antimicrobial activity against fish spoilage or pathogenic bacteria from the same host or ecological niche. The studied Cameroonian aquatic niche is an ideal source of antagonistic LAB that could be appropriate as new fish biopreservatives or disease control agents in aquaculture under tropical conditions in particular or worldwide in general.Graphical abstract

Effect of postharvest UV-C treatment on the bacterial diversity of Ataulfo mangoes by PCR-DGGE, survival of E. coli and antimicrobial activity

Since Mexico is the second largest exporter of mangoes, its safety assurance is essential. Research in microbial ecology and knowledge of complex interactions among microbes must be better understood to achieve maximal control of pathogens. Therefore, we investigated the effect of UV-C treatments on bacterial diversity of the Ataulfo mangoes surface using PCR-DGGE analysis of variable region V3 of 16S rRNA genes, and the survival of E. coli, by plate counting. The UV-C irradiation reduced the microbial load on the surface of mangoes immediately after treatment and the structure of bacterial communities was modified during storage. We identified the key members of the bacterial communities on the surface of fruits, predominating Enterobacter genus. Genera as Lactococcus and Pantoea were only detected on the surface of non-treated (control) mangoes. This could indicate that these genera were affected by the UV-C treatment. On the other hand, the treatment did not have a significant effect on survival of E. coli. However, genera that have been recognized as antagonists against foodborne pathogens were identified in the bands patterns. Also, phenolic compounds were determined by HPLC and antimicrobial activity was assayed according to the agar diffusion method. The main phenolic compounds were chlorogenic, gallic, and caffeic acids. Mango peel methanol extracts (UV-C treated and control mangoes) showed antimicrobial activity against strains previously isolated from mango, detecting significant differences (P < 0.05) among treated and control mangoes after 4 and 12 days of storage. Ps. fluorescens and Ps. stutszeri were the most sensitive.

Draft Genome Sequence of Leuconostoc citreum CW28 Isolated from Pozol, a Pre-Hispanic Fermented Corn Beverage

ABSTRACT Leuconostoc citreum CW28 was isolated from pozol, a Mayan fermented corn beverage. This strain produces a cell-associated inulosucrase, the first described in bacteria. Its draft genome sequence, announced here, has an estimated size of 1.98 Mb and harbors 1,915 coding genes, 12 rRNAs, 68 tRNAs, 17 putative pseudogenes, and 1 putative phage.