Microbiological, immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese containing Lactobacillus rhamnosus EM1107.

Cheeses are able to serve as suitable matrices for supplying probiotics to consumers, enabling appropriate conditions for bacteria to survive gastric transit and reach the gut, where they are assumed to promote beneficial processes. The present study aimed to evaluate the microbiological, immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese supplemented with the probiotic strain Lactobacillus rhamnosus EM1107. Thirty male albino Wistar rats were randomly distributed into 5 experimental groups with 6 animals each: negative (NC) and positive (PtC) control groups, control goat cheese (CCh), goat cheese added with L. rhamnosus EM1107 (LrCh), and L. rhamnosus EM1107 only (EM1107). All animals, except NC group were challenged with Salmonella Enteritidis (109 cfu in 1 mL of saline through oral gavage). Microbial composition was assessed with high-throughput 16S rRNA sequencing by means of Illumina MiSeq (Illumina, San Diego, CA). Nuclear factor kappa B (NF-κB) from the animal cecum tissue was determined by real-time PCR and interleukins (TNF-α, IL-1ß, IL-10, and IFN-γ) by means of ELISA. Myeloperoxidase and malondialdehyde levels were determined biochemically. The administration of the L. rhamnosus EM1107 probiotic strain, either as a pure culture or added to a cheese matrix, was able to reduce Salmonella colonization in the intestinal lumen and lessen tissue damage compared with rats from PtC group. In addition, the use of cheese for the probiotic strain delivery (LrCh) was associated with a marked shift in the gut microbiota composition toward the increase of beneficial organisms such as Blautia and Lactobacillus and a reduction in NF-κB expression. These findings support our hypothesis that cheeses might be explored as functional matrices for the efficacious delivery of probiotic strains to consumers.

Impact of pH and application time of meta-phosphoric acid on resin-enamel and resin-dentin bonding.

PURPOSE: To evaluate the immediate microshear resin-enamel bond strength (µSBS) and the immediate and 6-month microtensile bond strength (µTBS) and nanoleakage (NL) of the adhesive interface performed by different pHs of 40% meta-phosphoric acid (MPA) were compared with conventional 37% ortho-phosphoric acid (OPA) under different application times. Additionally, the enamel etching patterns were evaluated and the chemical/morphological changes induced by these differents groups were evaluated. MATERIALS AND METHODS: One hundred and ninety-eight extracted human molars were randomly assigned into experimental groups according to the combination of independent variables: Acid [37% ortho-phosphoric acid (OPA), 40% meta-phosphoric acid (MPA) at pHs of: 0.5, 1 and 2] and Application Time [7, 15 and 30s]. Enamel-bond specimens were prepared and tested under µSBS. Resin-dentin beams were tested under µTBS tested immediately or after 6-months of water storage. Nanoleakage was evaluated using bonded-beams of each tooth/time-period. Enamel etching pattern and chemical and ultra-morphology analyses were also performed. The µSBS (MPa) data were subjected to a two-way repeated measures ANOVA (Acid vs. Application time). For µTBS, Acid vs application time vs storage time data were subjected to three-way ANOVA and Tukey's test (α = 0.05). RESULTS: MPA pH 0.5 showed µTBS similar to OPA, independently of the application time on enamel (p>0.05) or dentin (p>0.05). OPA provided higher nanoleakage values than MPA (p = 0.003). Significant decreases in TBS and increases in NL were only observed for OPA after 6 months (p = 0.001). An increase in the application time resulted in a more pronounced etching pattern for MPA. Chemical analysis showed that dentin demineralized by MPA depicted peaks of brushite and octacalcium phosphate. MPA exposed less collagen than OPA. However, optimal results for MPA were dependent on pH/application time. CONCLUSION: The use of 40% meta-phosphoric acid with a pH of 0.5 is an alternative acid-etching agent for dentin and enamel bonding. Furthermore, the use of MPA preserves the resin-dentin interface over a 6-months period, due to presence of brushite and octacalcium phosphate and a reduced demineralization pattern.

Efficacy of new natural biomodification agents from Anacardiaceae extracts on dentin collagen cross-linking.

OBJECTIVES: Several polyphenols from renewable sources were surveyed for dentin biomodification. However, phenols from cashew nut shell liquid (CNSL, Anacardium occidentale) and from Aroeira (Myracrodruon urundeuva) extract have never been evaluated. The present investigation aimed to compare the dentin collagen crosslinking (biomodification) effectiveness of polyphenols from Aroeira stem bark extract, proanthocyanidins (PACs) from grape-seed extract (Vitis vinifera), cardol and cardanol from CNSL after clinically relevant treatment for one minute. METHODS: Three-point bending test was used to obtain the elastic modulus of fully demineralized dentin beams before and after biomodification, whilst color change and mass variation were evaluated after four weeks water biodegradation. Color aspect was assessed by optical images after biodegradation whereas collagen cross-linking was investigated by micro-Raman spectroscopy. Statistical analysis was performed with repeated-measures two way ANOVA and Tukey's test (p<0.05). RESULTS: The increase in elastic modulus after biomodification was in the order cardol>cardanol>aroeira=PACs with cardol solution achieving mean 338.2% increase. The mass increase after biomodification followed the same order aforementioned. Nevertheless, after four weeks aging, more hydrophobic agent (cardanol) induced the highest resistance against water biodegradation. Aroeira and cardol attained intermediate outcomes whereas PACs provided the lower resistance. Tannin-based agents (Aroeira and PACs) stained the specimens in dark brown color. No color alteration was observed with cardol and cardanol treatments. All four agents achieved crosslinking in micro-Raman after one minute application. SIGNIFICANCE: In conclusion, major components of CNSL yield overall best dentin biomodification outcomes when applied for one minute without staining the dentin collagen.

Efeito de diferentes aditivos zootécnicos sobre a qualidade de ovos em duas fases produtivas da codorna / Effect of different zootechnical additives on egg quality in two quail production phases

Com o objetivo de avaliar a qualidade dos ovos de codornas alimentadas com diferentes aditivos em duas fases de produção, 360 codornas foram distribuídas em um delineamento inteiramente ao acaso, em esquema de parcela subdividida no tempo (de nove até 23 e de 24 até 39 semanas de idade), com cinco tratamentos e oito repetições de nove aves cada, organizados da seguinte forma: controle, antibiótico, prebiótico, probiótico e simbiótico. As variáveis analisadas foram: unidade Haugh, índice de gema, porcentagem dos componentes do ovo, espessura de casca medida pelo micrômetro e pelo MEV. O fornecimento dos aditivos no segundo período aumentou a unidade Haugh e nos dois períodos avaliados proporcionou aumento da porcentagem de gema e de casca e da espessura da casca e de suas membranas. A inclusão dos aditivos foi mais eficiente em aumentar o peso dos ovos e a porcentagem de gema e de casca no segundo período. A espessura da casca (MEV) melhorou após inclusão de antibiótico e simbiótico na ração no segundo período. A inclusão de aditivos na ração melhora a qualidade dos ovos de codornas durante toda a fase produtiva. Antibiótico e simbiótico são mais eficazes em melhorar a espessura da casca dos ovos produzidos durante o segundo período produtivo.(AU)

To evaluate the quality of eggs of quails fed with different additives in two stages of the productive period, 360 quails were distributed in a completely randomized design in a split plot scheme in time (9 to 23 and 24 to 39 weeks of age) with five treatments and eight repetitions of nine birds each, organized as follows: control, antibiotic, prebiotic, probiotic, and symbiotic. The analyzed variables were: Haugh unit, yolk index, percentage of egg components, shell thickness measured by the micrometer and the SEM. The supply of additives in the second period increased Haugh units, and in both periods provided an increase in the percentage of yolk and shell and improved the thickness of the shell and its membranes. The inclusion of additives was more effective in promoting weight gain in eggs and the percentage of egg yolk and egg shell from 24 to 39 weeks of age. The shell thickness (SEM) improved after inclusion of antibiotic and symbiotic in feed in the second period. The addition of additives in food improved the quality of Japanese quail egg throughout the productive period. Antibiotics and symbiotics proved to be more effective in improving shell thickness of the eggs produced in the second productive period.(AU)

Efeito de diferentes aditivos zootécnicos sobre a qualidade de ovos em duas fases produtivas da codorna / Effect of different zootechnical additives on egg quality in two quail production phases

Com o objetivo de avaliar a qualidade dos ovos de codornas alimentadas com diferentes aditivos em duas fases de produção, 360 codornas foram distribuídas em um delineamento inteiramente ao acaso, em esquema de parcela subdividida no tempo (de nove até 23 e de 24 até 39 semanas de idade), com cinco tratamentos e oito repetições de nove aves cada, organizados da seguinte forma: controle, antibiótico, prebiótico, probiótico e simbiótico. As variáveis analisadas foram: unidade Haugh, índice de gema, porcentagem dos componentes do ovo, espessura de casca medida pelo micrômetro e pelo MEV. O fornecimento dos aditivos no segundo período aumentou a unidade Haugh e nos dois períodos avaliados proporcionou aumento da porcentagem de gema e de casca e da espessura da casca e de suas membranas. A inclusão dos aditivos foi mais eficiente em aumentar o peso dos ovos e a porcentagem de gema e de casca no segundo período. A espessura da casca (MEV) melhorou após inclusão de antibiótico e simbiótico na ração no segundo período. A inclusão de aditivos na ração melhora a qualidade dos ovos de codornas durante toda a fase produtiva. Antibiótico e simbiótico são mais eficazes em melhorar a espessura da casca dos ovos produzidos durante o segundo período produtivo.(AU)

To evaluate the quality of eggs of quails fed with different additives in two stages of the productive period, 360 quails were distributed in a completely randomized design in a split plot scheme in time (9 to 23 and 24 to 39 weeks of age) with five treatments and eight repetitions of nine birds each, organized as follows: control, antibiotic, prebiotic, probiotic, and symbiotic. The analyzed variables were: Haugh unit, yolk index, percentage of egg components, shell thickness measured by the micrometer and the SEM. The supply of additives in the second period increased Haugh units, and in both periods provided an increase in the percentage of yolk and shell and improved the thickness of the shell and its membranes. The inclusion of additives was more effective in promoting weight gain in eggs and the percentage of egg yolk and egg shell from 24 to 39 weeks of age. The shell thickness (SEM) improved after inclusion of antibiotic and symbiotic in feed in the second period. The addition of additives in food improved the quality of Japanese quail egg throughout the productive period. Antibiotics and symbiotics proved to be more effective in improving shell thickness of the eggs produced in the second productive period.(AU)

Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

Zika in Rio de Janeiro: Assessment of basic reproduction number and comparison with dengue outbreaks.

Zika virus infection was declared a public health emergency of international concern in February 2016 in response to the outbreak in Brazil and its suspected link with congenital anomalies. In this study, we use notification data and disease natural history parameters to estimate the basic reproduction number (R 0) of Zika in Rio de Janeiro, Brazil. We also obtain estimates of R 0 of dengue from time series of dengue cases in the outbreaks registered in 2002 and 2012 in the city, when DENV-3 and DENV-4 serotypes, respectively, had just emerged. Our estimates of the basic reproduction number for Zika in Rio de Janeiro based on surveillance notifications (R 0 = 2·33, 95% CI: 1·97-2·97) were higher than those obtained for dengue in the city (year 2002: R 0 = 1·70 [1·50-2·02]; year 2012: R 0 = 1·25 [1·18-1·36]). Given the role of Aedes aegypti as vector of both the Zika and dengue viruses, we also derive R 0 of Zika as a function of both dengue reproduction number and entomological and epidemiological parameters for dengue and Zika. Using the dengue outbreaks from previous years allowed us to estimate the potential R 0 of Zika. Our estimates were closely in agreement with our first Zika's R 0 estimation from notification data. Hence, these results validate deriving the potential risk of Zika transmission in areas with recurring dengue outbreaks. Whether transmission routes other than vector-based can sustain a Zika epidemic still deserves attention, but our results suggest that the Zika outbreak in Rio de Janeiro emerged due to population susceptibility and ubiquitous presence of Ae. aegypti.

Early menarche in normal-weight girls and its association with excess weight, abdominal obesity and metabolic changes at the end of sexual maturation.

BACKGROUND/OBJECTIVES: The objective of this study was to assess whether early menarche in normal-weight girls is associated with excess weight, abdominal obesity and metabolic changes at the end of sexual maturation. SUBJECTS/METHODS: This is a cohort study of 255 normal-weight girls aged 10-18 years attending public schools in the city of Recife, Pernambuco, Northeast of Brazil. The following data were collected in 2007 and 2012: anthropometric; lifestyle, socioeconomic and demographic data; lipid profile; and self-assessed sexual maturation. RESULTS: Eighty-four girls (32.9%) had early menarche. The socioeconomic level and time spent using a computer increased during this 5-year period, whereas the time spent watching television decreased. The prevalences of excess weight and abdominal obesity were 19.2% and 9.8%, respectively. The anthropometric and laboratory variables of girls with and without early menarche did not differ. CONCLUSIONS: These findings suggest that early menarche did not influence weight, abdominal obesity and serum lipids in the final stage of sexual maturation.

Shiga toxigenic and enteropathogenic Escherichia coli in water and fish from pay-to-fish ponds.

UNLABELLED: Escherichia coli is part of the normal microflora of the intestines of mammals. However, among the enteric pathogens, it is one of the leading causes of intestinal diseases, especially Shiga toxigenic E. coli, which can cause diarrhoea, haemorrhagic colitis and complications like haemolytic uraemic syndrome and thrombotic thrombocytopaenic purpura. Escherichia coli is considered a serious public health problem. Water and fish samples were subjected to biochemical tests to confirm the presence of E. coli and by PCR to verify the presence of pathogenic strains (O157, enteropathogenic and shiga toxigenic) in water and fish (skin, gastrointestinal tract and muscles) from pay-to-fish ponds located in the Córrego Rico watershed in the northeastern region of the state of São Paulo, Brazil. Of the 115 E. coli isolates from fish or water, five (4·34%) contained eae and stx2 genes, one had only the eae gene and two had the stx1 gene. An isolate containing the stx2 gene was also found in the water sample. In addition, eight isolates (6·95%) from the fish gastrointestinal tract contained rfbEO157:H7 (O157 gene), and three (2·61%) contained stx2 and eae genes, demonstrating the potential risk to the environment and public health. The results provide useful basic information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population. SIGNIFICANCE AND IMPACT OF THE STUDY: Pay-to-fish ponds are a common commercial activity in Brazil. Samples of water and Oreochromis niloticus were examined by PCR to detect the presence of pathogenic strains of Escherichia coli (O157, enteropathogenic and shiga toxigenic). Several pathogenic strains were detected in this study, providing useful epidemiological information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population.

Reaction norms for the study of genotype-environment interaction for growth and indicator traits of sexual precocity in Nellore cattle.

The objective of this study was to quantify the magnitude of genotype-environment interaction (GxE) effects on age at first calving (AFC), scrotal circumference (SC), and yearling weight (YW) in Nellore cattle using reaction norms. For the study, 89,152 weight records of female and male Nellore animals obtained at yearling age were used. Genetic parameters were estimated with a single-trait random-regression model using Legendre polynomials as base functions. The heritability estimates were of low to medium magnitude for AFC (0.05 to 0.47) and of medium to high magnitude for SC (0.32 to 0.51) and YW (0.13 to 0.72), and increased as the environmental gradient became more favorable. The genetic correlation estimates ranged from 0.25 to 1.0 for AFC, from 0.71 to 1.0 for SC, and from 0.42 to 1.0 for YW. High Spearman correlation coefficients were obtained for the three traits, ranging from 0.97 to 0.99. The reaction norms along the environmental gradient of 10 sires each with the highest or lowest breeding value for YW predicted by single-trait analysis demonstrated more plastic phenotypes for YW and more robust phenotypes for SC. The effect of GxE was most important for YW and AFC with respect to SC. When animals are selected for higher SC or YW or lower AFC, considering or not the GxE effect, it is expected that the same animals will be selected. The reaction norms obtained based on sire breeding values along the environmental gradient showed that animals with extreme breeding values respond differently as environmental conditions improve.

Alcanivorax dieselolei, an alkane-degrading bacterium associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa).

Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.

Genotype × environment interaction for age at first calving, scrotal circumference, and yearling weight in Nellore cattle using reaction norms in multitrait random regression models.

The aim of this study was to evaluate the effect of genotype × environment interaction (G×E) on age at first calving (AFC), scrotal circumference (SC), and yearling weight (YW) and to estimate genetic correlations between these traits in Nellore cattle using reaction norms in multitrait random regression models. In this study, 28,871, 41,386, and 89,152 records of Nellore cattle for AFC, SC, and YW, respectively, were used. The data were obtained from farms located in the north, northeast, midwest, and southeast regions of Brazil that participate in the DeltaGen Breeding Program. Environmental levels were defined as a function of contemporary groups, that is, animals born in the same herd and year, from the same management group (from birth to yearling), and of the same sex. Postweaning weight gain was used as a criterion to evaluate the environmental conditions for all traits. For reaction norm analyses, residual variances were modeled with homogeneous and heterogeneous classes. The model for SC and YW included the fixed effects of contemporary group and age of the animal as a covariate as well as random direct additive genetic and residual effects. The same model, excluding the covariate age of the animal, was used for AFC. The heritability estimates were low to high for AFC (0.09 to 0.50), high for SC (0.51 to 0.67), and moderate to high for YW (0.33 to 0.71). The genetic correlations (within each trait) along the environmental levels varied from -0.27 to 1.0 for AFC, from 0.73 to 1.0 for SC, and from 0.26 to 1.0 for YW. The genetic correlations between different traits in different environments varied from -0.14 to -0.60 between AFC and SC, from -0.05 to -0.32 between AFC and YW, and from -0.05 to 0.72 between YW and SC. The genetic correlations have had different magnitudes for AFC, SC, and YW, which could indicate the presence of G×E. The present results should support researchers and farmers in defining selection criteria to improve growth traits and sexual precocity. Our results suggest that animals for breeding have to be selected in the same environment and management conditions as their progeny will be reared.

Alcanivorax dieselolei, an alkane-degrading bacterium associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa) / Alcanivorax dieselolei, uma bactéria degradadora de alcanos associada ao muco do zoantídeo Palythoa caribaeorum (Cnidaria, Anthozoa)

Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.(AU)

Análises dos genes 16S rDNA foram empregadas para identificar a microbiota isolada do muco do zoantídeo Palythoa caribaeorum de Porto de Galinhas, litoral do estado de Pernambuco, Brasil. Este estudo é importante pelo ineditismo dessa associação, pelas relevantes aplicações biotecnológicas da bactéria Alcanivorax dieselolei e pela indicação da presença de uma bactéria degradadora de hidrocarbonetos em um ecossistema recifal como o de Porto de Galinhas.(AU)

Alcanivorax dieselolei, an alkane-degrading bacterium associated with the mucus of the zoanthid Palythoa caribaeorum (Cnidaria, Anthozoa) / Alcanivorax dieselolei, uma bactéria degradadora de alcanos associada ao muco do zoantídeo Palythoa caribaeorum (Cnidaria, Anthozoa)

Analyses of 16S rDNA genes were used to identify the microbiota isolated from the mucus of the zoanthid Palythoa caribaeorum at Porto de Galinhas on the coast of Pernambuco State, Brazil. This study is important as the first report of this association, because of the potential biotechnological applications of the bacterium Alcanivorax dieselolei, and as evidence for the presence of a hydrocarbon degrading bacterium in a reef ecosystem such as Porto de Galinhas.

Análises dos genes 16S rDNA foram empregadas para identificar a microbiota isolada do muco do zoantídeo Palythoa caribaeorum de Porto de Galinhas, litoral do estado de Pernambuco, Brasil. Este estudo é importante pelo ineditismo dessa associação, pelas relevantes aplicações biotecnológicas da bactéria Alcanivorax dieselolei e pela indicação da presença de uma bactéria degradadora de hidrocarbonetos em um ecossistema recifal como o de Porto de Galinhas.

Fate of non O157 Shigatoxigenic Escherichia coli in ovine manure composting / Eliminação de Escherichia coli Shigatoxigênica não O157 em compostagem de esterco ovino

Livestock manure may contain pathogenic microorganisms which pose a risk to the health of animal or humans if the manure is not adequately treated or disposed of. To determine the fate of Shiga toxigenic Escherichia coli (STEC) non O157 in composted manure from naturally colonized sheep, fresh manure was obtained from animals carrying bacterial cells with stx1/ stx2 genes. Two composting systems were used, aerated and non-aerated, and the experiments were done in Dracena city, São Paulo State. Every week, for seven weeks, one manure sample from six different points in both systems was collected and cultured to determine the presence of E. coli, the presence of the virulence genes in the cells, and also the susceptibility to 10 antimicrobial drugs. The temperature was verified at each sampling. STEC non-O157 survived for 49 days in both composting systems. E. coli non-STEC showing a high degree of antibiotic resistance was recovered all long the composting period. No relationship was established between the presence of virulence genes and antibiotic resistance. The presence of virulence genes and multiple antibiotic resistances in E. coli implicates a potential risk for these genes spread in the human food chain, which is a reason for concern.(AU)

Esterco de animais de criação pode conter microrganismos patogênicos, o que representa um risco para a saúde animal e a humana se o esterco não for adequadamente tratado ou descartado. Determinou-se o tempo necessário para a eliminação de Escherichia coli Shiga toxigenica (STEC) não O157 em esterco ovino composto, obtido de fezes frescas de ovelhas naturalmente colonizadas com cepas STEC não O157 que apresentavam os genes stx1/ stx2. Foram utilizados dois sistemas de compostagem, aerado e não aerado, em experimentos realizados na cidade de Dracena, estado de São Paulo. Todas as semanas, durante sete semanas, uma amostra de compostagem proveniente de seis pontos diferentes na leira, nos dois sistemas, foi coletada e semeada para a determinação da presença de E. coli, da presença de genes de virulência nas células, bem como da sensibilidade dessas células a 10 drogas antimicrobianas. Em cada amostragem, a temperatura da leira foi analisada. Células de STEC não O157 sobreviveram por 49 dias nos dois sistemas de compostagem. E. coli não STEC com um alto grau de resistência a antibióticos foi recuperada ao longo de todo o período de compostagem. Não foi possível estabelecer relação entre a presença de genes de virulência e a resistência a antibióticos. A presença de genes de virulência e a resistência a múltiplos antibióticos em E. coli representam um risco potencial para o espalhamento desses genes na cadeia alimentar humana, o que é motivo de grande preocupação.(AU)

Fate of non O157 Shigatoxigenic Escherichia coli in ovine manure composting / Eliminação de Escherichia coli Shigatoxigênica não O157 em compostagem de esterco ovino

Livestock manure may contain pathogenic microorganisms which pose a risk to the health of animal or humans if the manure is not adequately treated or disposed of. To determine the fate of Shiga toxigenic Escherichia coli (STEC) non O157 in composted manure from naturally colonized sheep, fresh manure was obtained from animals carrying bacterial cells with stx1/ stx2 genes. Two composting systems were used, aerated and non-aerated, and the experiments were done in Dracena city, São Paulo State. Every week, for seven weeks, one manure sample from six different points in both systems was collected and cultured to determine the presence of E. coli, the presence of the virulence genes in the cells, and also the susceptibility to 10 antimicrobial drugs. The temperature was verified at each sampling. STEC non-O157 survived for 49 days in both composting systems. E. coli non-STEC showing a high degree of antibiotic resistance was recovered all long the composting period. No relationship was established between the presence of virulence genes and antibiotic resistance. The presence of virulence genes and multiple antibiotic resistances in E. coli implicates a potential risk for these genes spread in the human food chain, which is a reason for concern...

Esterco de animais de criação pode conter microrganismos patogênicos, o que representa um risco para a saúde animal e a humana se o esterco não for adequadamente tratado ou descartado. Determinou-se o tempo necessário para a eliminação de Escherichia coli Shiga toxigenica (STEC) não O157 em esterco ovino composto, obtido de fezes frescas de ovelhas naturalmente colonizadas com cepas STEC não O157 que apresentavam os genes stx1/ stx2. Foram utilizados dois sistemas de compostagem, aerado e não aerado, em experimentos realizados na cidade de Dracena, estado de São Paulo. Todas as semanas, durante sete semanas, uma amostra de compostagem proveniente de seis pontos diferentes na leira, nos dois sistemas, foi coletada e semeada para a determinação da presença de E. coli, da presença de genes de virulência nas células, bem como da sensibilidade dessas células a 10 drogas antimicrobianas. Em cada amostragem, a temperatura da leira foi analisada. Células de STEC não O157 sobreviveram por 49 dias nos dois sistemas de compostagem. E. coli não STEC com um alto grau de resistência a antibióticos foi recuperada ao longo de todo o período de compostagem. Não foi possível estabelecer relação entre a presença de genes de virulência e a resistência a antibióticos. A presença de genes de virulência e a resistência a múltiplos antibióticos em E. coli representam um risco potencial para o espalhamento desses genes na cadeia alimentar humana, o que é motivo de grande preocupação...

Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.

The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.

Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana.

Qualidade microbiológica da carcaça bovina durante o processo de abate e a ocorrência de e. Coli o157 h7 na carne / Microbiological quality of cattle carcass during harvest process and the occurrence of e. Coli o157 h7 in meat

A Escherichia coli O157: H7 é uma importante cepa associada a surtos graves de enfermidade em seres humanos, a maioria deles derivada do consumo de carne crua ou mal cozida. É provável que o gado atue como um importante reservatório, sugerindo-se a possibilidade de que a gestão da dieta no confinamento possa influenciar o aparecimento de cepas Shigatoxigênicas. Este estudo teve como objetivo verificar a qualidade microbiológica das carcaças e a ocorrência de E. coli O157: H7, por meio dos resultados obtidos por métodos indicadores (contagem total de microrganismos viáveis, contagem de Coliformes e de E. coli) e por um método automatizado de PCR para detecção de E. coli O157: H7. Foram colhidas amostras de retalhos de carne (carne industrial) e de carcaças de bovinos terminados em pastagem ou em confinamento, permitindo o fornecimento de subsídios necessários para a Análise de Perigos e Pontos Críticos de Controle (HACCP). Desses mesmos animais foram colhidas, também, amostras de swab retal para a detecção experimental de E. coli O157: H7 nas fezes. Um total de 100 swabs retais, 100 amostras de carcaças quentes, além de outras 323 amostras de aparas de carne (retalhos da desossa), foram analisados. Com exceção de uma amostra de retalhos de carne (0,31%), todas as demais, de fezes e de carcaças, foram negativas para a presença de E. coli O157: H7. Não houve diferenças significativas entre os tipos de terminação utilizada para o gado. Os resultados dos métodos indicadores foram considerados aceitáveis em 91%, 85% e 93% das amostras testadas, respectivamente, para a CTV, contagem de Coliformes e de E. coli de carcaças, dando suporte e em acordo com a baixa ocorrência da cepa O157: H7

Escherichia coli O157:H7, an important bacillus strain associated with serious gastroenteritis in humans, is more frequently derived from the consumption of raw or poorly cooked beef. Cattle are important reservoirs suggesting the possibility that feedlot diet management influences the emergence of Shiga-toxigenic strains. This study evaluates the microbiological quality of carcasses and the occurrence of E. coli O157:H7 using the results from general indicator methods (total viable count, coliform rate and E. coli counts) and by an automated PCR method for the detection of E. coli O157:H7. Samples were collected from (industrially processed) meat trimmings and from carcasses of cattle finished on pasture or in feedlots so that sufficient data for the Hazard Analysis and Critical Control Points (HACCP) could be obtained. Samples of rectal swab for experimental detection of E. coli O157:H7 were also collected. One hundred rectal swabs, 100 samples retrieved from warm carcasses and 323 samples of meat trimmings were analyzed. With the exception of one sample of meat trim (0.31%), all the other samples from excreta and carcasses were negative for the O157:H7 E. coli strain. There were no significant differences between the methods used for cattle finishing. Indicator methods results were considered acceptable in 91%, 85% and 93% of tested samples of carcasses respectively for TVC, coliform and E. coli counts. These results agree with statistical data showing the low occurrence of O157:H7 strain