High accumulation of perfluorooctane sulfonate (PFOS) in marine tucuxi dolphins (Sotalia guianensis) from the Brazilian coast

Perfluorooctane sulfonate (PFOS) and other perfluoroalkyl compounds (PFCs) were measured in liver samples from 29 marine tucuxi dolphins from Rio de Janeiro state (RJ), Brazil. PFC measurement combined liquid chromatography and mass spectrometry, using a CapLC system connected to a Quadrupole-LIT mass spectrometer. PFOS was the only PFC detected and it was so in all samples. PFOS concentrations (ng x g(-1) dw) of dolphins (n=23) from the highly contaminated Guanabara Bay (in RJ) varied between 43 and 2431 as well as between 76 and 427 from areas of RJ other than the quoted bay (n=6). Concentrations of three fetuses and one neonate varied between 664 and 1590. Fetus/mother ratios were calculated in two situations (2.75 and 2.62). It seems that mother-to-calf transference plays important role for relationships between PFOS and age. When a one-year-old male calf presenting 2431 ng x g(-1) dw was excluded from the test, significant correlations were observed between PFOS concentrations and both age and total length. Despite the placental transference, PFOS concentrations were not significantly lower in females than in males. PFOS levels in marine tucuxi dolphins from Guanabara Bay are among the highest detected to date in cetaceans, and this may represent a threat to the small population concerned.     

Identification of tet(M) in two Lactococcus lactis strains isolated from a Spanish traditional starter-free cheese made of raw milk and conjugative transfer of tetracycline resistance to lactococci and enterococci

Specific PCR and sequencing showed that a tet(M) gene was present in two tetracycline-resistant Lactococcus lactis strains isolated from a raw milk, starter-free cheese. Hybridisation experiments using as a probe an internal segment of the gene obtained by PCR associated tet(M) with plasmids of around the same size (30 kbp) in both strains. Molecular analysis of the tetracycline resistance loci, including the upstream and downstream regions of the genes, showed them to be identical to one other and to the tet(M) encoded by the conjugative transposon Tn916. Amplification of Tn916-derived segments suggested the transposon was complete in the two L. lactis strains. Further, curing of the tetracycline resistance was accompanied by a reduction in size of the plasmids comparable to that expected for Tn916. Tetracycline resistance could be transferred by conjugation to plasmid-free Lactococcus and Enterococcus strains. However, no plasmid DNA was detected among the transconjugants while both tet(M) and transposon-related sequences were amplified by PCR. This suggested that only the transposon was mobilized.     

Spoilage of blood sausages morcilla de Burgos treated with high hydrostatic pressure

In this study, the microbial ecology of the blood sausage morcilla de Burgos, subjected to high hydrostatic pressure treatment (HPP), was studied by culture-dependent and -independent methods. Morcilla de Burgos is the most traditional and famous blood sausage in Spain. The producers are interested in extending its shelf-life in order to expand their market and to reduce losses attributed to spoilage. Sausage batter prior to stuffing and blood sausages HPP treated or not (control) were analyzed at 0, 9, 14, 21, 28 and 35 days of storage at 4 degrees C. Lactic acid bacteria, Pseudomonas spp. and aerobic mesophilic bacteria were investigated by traditional plating. PCR-denaturing gradient gel electrophoresis (DGGE) was used to analyze the DNA and the RNA extracted directly from the blood sausages, as well as bulk cells of LAB and Pseudomonas spp. The results showed that HPP improved the shelf life of morcilla de Burgos to 28 days in comparison with control samples. The populations responsible for spoilage, namely LAB, remained lower in HPP treated samples when compared with the control samples. Only at 35 days of storage they reached values of 10(8) cfu/g, leading to the spoilage of the product. Although, HPP affected the LAB population, they were able to recover the injury provoked by the treatment. Lastly, HPP seemed to affect differently LAB species detected. While Leuconostoc mesenteroides was completely inactivated by HPP, Weissella viridescens was able to recover and carry out the typical spoilage of the product. Pseudomonas spp. remained under detection level (<10(2) CFU/g) after the HPP treatment.     

Modifications in membrane fatty acid composition of Salmonella typhimurium in response to growth conditions and their effect on heat resistance

The effects of growth temperature (in the range 10-45 degrees C) and acidification up to pH 4.5 of the culture medium (Brain Heart Infusion, BHI) with different organic acids (acetic, citric and lactic) and hydrochloric acid on membrane fatty acid composition and heat resistance of Salmonella typhimurium CECT 443 were studied. The heat resistance was maximal in cells grown at 45 degrees C (cells grown in non-acidified BHI showed a D58-value of 0.90 min) and decreased with decreasing growth temperature up to 10 degrees C (D58-value of 0.09 min). The growth of cells in acidified media caused an increase in their heat resistance. In general, acid adapted cells showed D-values of between 1.5 and 2 times higher than the corresponding for non-acid adapted control cells. This cross-protection response, which has important implications in food processing, was not dependent on the pH value and the acid used to acidify the growth medium. A membrane adaptation corresponding to an increase in the unsaturated to saturated fatty acids ratio (UFA/SFA) and membrane fluidity was observed at low growth temperature. Moreover, the acidification of the growth medium caused a decrease in UFA/SFA ratio and in the C18:1 relative concentration, and an increase in cyclopropane fatty acids (CFA) content mainly due to the increase in cyc19 relative concentration. Thus, acid adapted cells showed CFA levels 1.5 times higher than non-acid adapted control cells. A significant proportion of unsaturated fatty acids were converted to their cyclopropane derivatives during acid adaptation. These changes in membrane fatty acid composition result in cells with decreased membrane fluidity. A clear relation between membrane fatty acid composition and heat resistance was observed. In general, D-values were maximum for cells with low UFA/SFA ratio, and, consequently, with low membrane fluidity. Moreover, CFA formation played a major role in protecting acid adapted cells from heat inactivation. However, changes observed in membrane fatty acid composition are not enough to explain the great thermotolerance of cells grown at 45 degrees C. Thus, other mechanisms, such as the synthesis of Heat Shock Proteins, could be responsible for this increase in the bacterial heat resistance.     

PCR method for detection and identification of Lactobacillus casei/paracasei bacteriophages in dairy products

Bacteriophage infections of starter lactic acid bacteria (LAB) pose a serious risk to the dairy industry. Nowadays, the expanding use of valuable Lactobacillus strains as probiotic starters determines an increase in the frequency of specific bacteriophage infections in dairy plants. This work describes a simple and rapid Polymerase Chain Reaction (PCR) method that detects and identifies bacteriophages infecting Lactobacillus casei/paracasei, the main bacterial species used as probiotic. Based on a highly conserved region of the NTP-binding genes belonging to the replication module of L. casei phages phiA2 and phiAT3 (the only two whose genomes are completely sequenced), a pair of primers was designed to generate a specific fragment. Furthermore, this PCR detection method proved to be a useful tool for monitoring and identifying L. casei/paracasei phages in industrial samples since specific PCR signals were obtained from phage contaminated milk (detection limit: 10(4) PFU/mL milk) and other commercial samples (fermented milks and cheese whey) that include L. casei/paracasei as probiotic starter (detection limit: 10(6) PFU/mL fermented milk). Since this method can detect the above phages in industrial samples and can be easily incorporated into dairy industry routines, it might be readily used to earmark contaminated milk for use in processes that do not involve susceptible starter organisms, or processes which involve phage-deactivating conditions.     

The occurrence of fungi, yeasts and bacteria in the air of a Spanish winery during vintage

This research studies the presence of microorganisms of enological interest (yeasts, bacteria and molds) and their evolution in the air of a wine cellar. The samples were taken throughout the winemaking campaign (September-December) in a winery of the D.O.Ca. Rioja, Spain. They were collected using an airIDEAL atmosphere sampler from Biomerieux. For the isolation, specific selective media were used for each group of microorganisms. The results obtained indicate that the presence in the winery air of the various different microorganisms studied is directly related to the winemaking processes that are taking place in the winery. Thus, the number of molds present decreases once grapes have ceased to be brought into the winery. The maximum number of yeasts in the air is found when all the vats in the cellar are fermenting, while the lactic bacteria are not detected until the first malolactic fermentation begins. The species of yeasts and molds identified are also related to the winemaking processes. The coincidence of strains of Saccharomyces cerevisiae among those present in the vats during alcoholic fermentation and those isolated from the air, confirms the role of the latter as a transmitter of microorganisms.     

Nutritional status and its correlates in Equatorial Guinean preschool children: results from a nationally representative survey

BACKGROUND: In Equatorial Guinea, as a result of the recent growth of the oil industry, there is an opportunity to address important public health problems through public and private initiatives. To propose effective nutrition and public health strategies, it is important first to have reliable information on the nutritional status of the population and the underlying factors affecting it. OBJECTIVE: To assess the nutritional status and the prevalence of anemia among Equatoguinean children in a nationally representative sample and to identify the risk factors associated with the nutritional problems detected. METHODS: The study was a cross-sectional survey using a multistaged, stratified, cluster-selected sample. The survey included a sociodemographic, health, and dietary questionnaire and measurement of hematocrit and anthropometric features, from which nutritional indicators based on the National Center for Health Statistics (NCHS) reference and the World Health Organization (WHO) standards were calculated. Logistic regression models were used for the multivariate analysis. A total of 552 children aged 0 to 60 months were surveyed. RESULTS: The overall prevalence of stunting (< -2 height-for-age z-scores [HAZ]) was 29.7% based on the NCHS reference and 35.2% based on WHO standards; the risk factors associated with stunting were age (p < .0001), low socioeconomic status (p = .01), and fishing by a member of the household (p = .003) The prevalence of mild anemia (hemoglobin < 110 g/L) was 69.3%, and that of moderate or severe anemia (hemoglobin < 80 g/L) was 8.3%. The only significant risk factor associated with moderate to severe anemia was low household socioeducational level (p = .01). CONCLUSIONS: Stunting and anemia are public health problems in Equatorial Guinea. Integrated strategies, including fighting poverty and improving maternal education, should be undertaken.