Neutral Genomic Microevolution of a Recently Emerged Pathogen, Salmonella enterica Serovar Agona

Salmonella enterica serovar Agona has caused multiple food-borne outbreaks of gastroenteritis since it was first isolated in 1952. We analyzed the genomes of 73 isolates from global sources, comparing five distinct outbreaks with sporadic infections as well as food contamination and the environment. Agona consists of three lineages with minimal mutational diversity: only 846 single nucleotide polymorphisms (SNPs) have accumulated in the non-repetitive, core genome since Agona evolved in 1932 and subsequently underwent a major population expansion in the 1960s. Homologous recombination with other serovars of S. enterica imported 42 recombinational tracts (360 kb) in 5/143 nodes within the genealogy, which resulted in 3,164 additional SNPs. In contrast to this paucity of genetic diversity, Agona is highly diverse according to pulsed-field gel electrophoresis (PFGE), which is used to assign isolates to outbreaks. PFGE diversity reflects a highly dynamic accessory genome associated with the gain or loss (indels) of 51 bacteriophages, 10 plasmids, and 6 integrative conjugational elements (ICE/IMEs), but did not correlate uniquely with outbreaks. Unlike the core genome, indels occurred repeatedly in independent nodes (homoplasies), resulting in inaccurate PFGE genealogies. The accessory genome contained only few cargo genes relevant to infection, other than antibiotic resistance. Thus, most of the genetic diversity within this recently emerged pathogen reflects changes in the accessory genome, or is due to recombination, but these changes seemed to reflect neutral processes rather than Darwinian selection. Each outbreak was caused by an independent clade, without universal, outbreak-associated genomic features, and none of the variable genes in the pan-genome seemed to be associated with an ability to cause outbreaks. © 2013 Achtman et al

Bisphenol A migration from plastic materials: direct insight of ecotoxicity in Daphnia magna

Bisphenol A (BPA) is an endocrine disrupting chemical (EDC) whose migration from food packaging is recognized worldwide. However, the real overall food contamination and related consequences are yet largely unknown. Among humans, children’s exposure to BPA has been emphasized because of the immaturity of their biological systems. The main aim of this study was to assess the reproductive impact of BPA leached from commercially available plastic containers used or related to child nutrition, performing ecotoxicological tests using the biomonitoring species Daphnia magna. Acute and chronic tests, as well as single and multigenerational tests were done. Migration of BPA from several baby bottles and other plastic containers evaluated by GC-MS indicated that a broader range of foodstuff may be contaminated when packed in plastics. Ecotoxicological test results performed using defined concentrations of BPA were in agreement with literature, although a precocious maturity of daphnids was detected at 3.0 mg/L. Curiously, an increased reproductive output (neonates per female) was observed when daphnids were bred in the polycarbonate (PC) containers (145.1±4.3 % to 264.7±3.8 %), both in single as in multigenerational tests, in comparison with the negative control group (100.3±1.6 %). A strong correlated dose-dependent ecotoxicological effect was observed, providing evidence that BPA leached from plastic food packaging materials act as functional estrogen in vivo at very low concentrations. In contrast, neonate production by daphnids cultured in polypropylene and non-PC bottles was slightly but not significantly enhanced (92.5±2.0 % to 118.8±1.8 %). Multigenerational tests also revealed magnification of the adverse effects, not only on fecundity but also on mortality, which represents a worrying trend for organisms that are chronically exposed to xenoestrogens for many generations. Two plausible explanations for the observed results could be given: a non-monotonic dose–response relationship or a mixture toxicity effect.

Quantification of clenbuterol at trace level in human urine by ultra-high pressure liquid chromatography-tandem mass spectrometry.

Clenbuterol is a β2 agonist agent with anabolic properties given by the increase in the muscular mass in parallel to the decrease of the body fat. For this reason, the use of clenbuterol is forbidden by the World Anti-Doping Agency (WADA) in the practice of sport. This compound is of particular interest for anti-doping authorities and WADA-accredited laboratories due to the recent reporting of risk of unintentional doping following the eating of meat contaminated with traces of clenbuterol in some countries. In this work, the development and the validation of an ultra-high pressure liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method for the quantification of clenbuterol in human urine is described. The analyte was extracted from urine samples by liquid-liquid extraction (LLE) in basic conditions using tert butyl-methyl ether (TBME) and analyzed by UHPLC-MS/MS with a linear gradient of acetonitrile in 9min only. The simple and rapid method presented here was validated in compliance with authority guidelines and showed a limit of quantification at 5pg/mL and a linearity range from 5pg/mL to 300pg/mL. Good trueness (85.8-105%), repeatability (5.7-10.6% RSD) and intermediate precision (5.9-14.9% RSD) results were obtained. The method was then applied to real samples from eighteen volunteers collecting urines after single oral doses administration (1, 5 and 10μg) of clenbuterol-enriched yogurts.

Seven years of gamma-ray spectrometry interlaboratory comparisons in Switzerland.

Since the 1990s, regular comparisons of gamma-ray spectrometry in Switzerland were organized to improve laboratory abilities to measure the radioactivity in the environment and food stuffs at typical routine levels. The activity concentration of the test samples and the evaluation of the associated uncertainties remained each year the main required test result. Over the years, the comparisons used certified reference solutions as well as environmental samples. The aim of this study is to research the effect of the comparisons on measurement quality. An analysis of the seven last interlaboratory comparisons revealed that the Swiss measurement capability is up to date. In addition, the results showed that the participants now have an improved evaluation of the uncertainties associated with their measurement.

Arsenic accumulation and speciation in Cherax destructor

Yabbies from contaminated sites and those fed arsenic in a lab. study accumulated significant levels of arsenic mostly as As(III) and As(V). Arsenic accumulation affects the structure of the internal organs and contractile function of their muscle. The arsenic in their tissues was found to be bioaccessible to human consumers.

Ocorrência de Staphylococcus aureus em queijo tipo frescal

OBJETIVO: Verificar a ocorrência de Staphylococcus aureus em uma amostra de queijo tipo Minas frescal comercializado na cidade de Poços de Caldas, MG, de modo a obter subsídios que permitam avaliar o risco potencial que este produto pode representar para a saúde da população consumidora. MÉTODOS: Foi investigada a presença e o número de cepas de Staphylococcus aureus em 80 amostras de queijo tipo Minas frescal produzido artesanalmente e comercializado na cidade de Poços de Caldas, MG, Brasil. RESULTADOS: Os resultados obtidos evidenciaram a presença de S. aureus em 40 (50,0%) amostras, cujas contagens revelaram valores médios em torno de 10(5)/g. CONCLUSÕES: Tais achados parecem ser extremamente preocupantes, pois além de se situarem acima do limite máximo de 10³/g estabelecido pelo Ministério da Saúde, estes valores mostraram-se muito próximos dos requeridos para a produção de enterotoxinas em quantidades suficientes para a ocorrência de surtos de intoxicação alimentar estafilocócica.

Barium uptake by maize plants as affected by sewage sludge in a long-term field study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Exame microbiológico de sorvetes não pasteurizados

Realizou-se exame microbiológico em 24 amostras de sorvetes não pasteurizados, todos preparados de maneira não industrial e à base de leite (creme, nata, chocolate), fabricados por 12 sorveterias diferentes da cidade de Araraquara, SP. Colheram-se duas amostras de cada sorveteria com intervalo de 15 dias entre as colheitas. Realizaram-se as seguintes provas: contagem de bactérias aeróbicas ou facultativas mesófilas e psicrófilas e de Staphylococcus aureus; determinação do Número Mais Provável (NMP) de coliformes totais e fecais e da presença de Salmonella. As técnicas utilizadas foram aquelas convencionalmente usadas para tais determinações. Não foi encontrada Salmonella em nenhuma das amostras e de cerca de 16,6% delas isolou-se Staphylococcus aureus. em proporções variáveis verificou-se a presença de microrgarnismos deteriorantes e daqueles indicadores de poluição de origem fecal.

Atividade inseticida das plantas e aplicações: revisão

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bioaccumulation of Microcystins In Lettuce

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

CONDICOES BACTERIOLOGICAS DE LEITE CRU E DE LEITE PASTEURIZADO NA REGIAO DE ARARAQUARA, SP, BRASIL

Milk can be an important spreading vehicle of pathogenic agents mainly for young children who are an important group of milk consumers. 135 milk samples (77 of raw milk and 58 of pasteurized milk) were analysed in order to verify the number of heterotrophic bacteria, of Staphylococcus aureus and of total coliforms, as well as to determine presence of Salmonella, Shigella, enteropathogenic Escherichia coli (EPEC) and enteroinvasive Escherichia coli (EIEC). The results were negative for Salmonella, Shigella and EIEC. EPEC serotypes 0:28, 0:111 and 0:55 were isolated in 4 of raw milk samples. The heterotrophic bacteria was found in counts over 30.000 UFC/mL in 91%, 25% and 68,75% respectively of raw milk, milk grade B and grade C. Counts over 30.000 coliforms/mL were found in 70,13%, 6,25% and 6,24% of the raw milk, B and C, respectively. 32,40% of the raw milk had counts of S. aureus over 3.000 UFC/mL.

HPLC determination of flumethrin, deltamethrin, cypermethrin, and cyhalothrin residues in the milk and blood of lactating dairy cows

A procedure to determine residue concentrations of synthetic pyrethroid insecticides (flumethrin, deltamethrin, cypermethrin and cyhalothrin) in the milk and blood of lactating dairy cows was developed. Extraction was performed with acetonitrile, n-hexane partitioning, and silica gel column cleanup with n-hexane and diethyl ether. Analysis was carried out by high- performance liquid chromatography and ultraviolet detection. Recovery of the four pyrethroids averaged 78 to 91% with a minimum detectable concentration of 0.001 mg/kg. The method was reproducible and sensitive.

Incidence of patulin in fruits and fruit juices marketed in Campinas, Brazil

One hundred and eleven samples of processed fruit juices (apple, grape, pineapple, papaya, guava, banana and mango) and 38 samples of sound fruits (apple, papaya, mango, pear and peach) produced and marketed in Brazil, were analysed for patulin by HPLC. Only one out of 30 samples of apple juice was found positive at 17 μg/l. Patulin was not detected in the other foodstuffs. It was found in 14 samples of spoiled fruit samples of apple (150-267 μg/kg), pear (134-245 μg/kg) and peach (92-174 μg/kg). Confirmation of the identity of patulin was based on the UV spectrum obtained by the HPLC diode array detector, compared with that of standard patulin, TLC developed by several solvent systems and sprayed with 3-methyl-2-benzothiazolinone hydrazone, and by acetylation with acetic anhydride.