Uma análise histórico-crítica sobre o desenvolvimento das normas brasileiras relacionadas a queijos artesanais / [A critical-historical analysis of the continuous development of Brazilian legislation related to artisanal cheeses]

Diferentes tipos de queijos artesanais são produzidos, comercializados e consumidos no Brasil, o que impulsiona o constante desenvolvimento de normas por órgãos oficiais, como o Mapa. A criação do Suasa e do Sisbi-POA foi fundamental para esse setor, por permitir um sistema de equivalência na fiscalização e por ampliar a distribuição. Ainda, o Mapa passou a permitir que queijos artesanais produzidos com leite cru pudessem ser maturados em um período inferior a 60 dias, desde que comprovada sua inocuidade. A redução do tempo de maturação é um tema controverso e polêmico, já que não há critérios específicos que estudos científicos devem contemplar, o que permite múltiplas interpretações de dados. Com a criação e a regulamentação do selo Arte, a fiscalização dos produtos artesanais foi designada aos órgãos de agricultura, pecuária e de saúde pública, em complementação à atribuição já prevista pelo Mapa e pelo Sisbi-POA. Ainda, o selo Arte atribui aos órgãos de inspeção uma função orientadora, atividade que deveria ser prioritariamente executada por agências de extensão e associações. As normas que balizam a produção e comercialização de produtos artesanais devem ser frequentemente atualizadas, devido aos constantes avanços científicos na área e para assegurar a oferta de produtos com qualidade e inócuos aos consumidores.(AU)

Different artisanal cheeses are produced, commercialized and consumed in Brazil, leading to a constant development of related rules by the MAPA and other official agencies. The establishment of two national programs (SUASA and SISBI-POA) allowed an equivalence in inspection system and an expanded distribution. Also, MAPA allowed ripening time lower than 60 days for artisanal raw milk cheeses, based on scientific studies that assure their safety. However, lowering the ripening period is still controversial, once there are no proper established criteria for such scientific studies, leading to potential multiple interpretation of data. The newly established ARTE certification transferred the inspection responsibilities of artisanal products to secretaries of agriculture, livestock and health, in support of what was already predicated by MAPA and SISBI-POA. Based on ARTE certification, the inspection service must also provide orientation guidance to producers, which should be done specifically by extension organs and associations. The norms that guide the production and commercialization of these artisanal products often need to be updated, but based on well-established methodologies and procedures, to ensure the distribution of suitable products to consumers.(AU)

Association between ideal cardiovascular health and depression incidence: a longitudinal analysis of ELSA-Brasil.

OBJECTIVE: We investigated whether ideal cardiovascular health (ICH), a metric proposed by the American Heart Association, predicts depression development. METHODS: Cohort analysis from the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil). Adults with no current depression and other common mental disorders, cardiovascular diseases, and antidepressant drug use at baseline had their ICH (composite score of smoking, dietary habits, body mass index, blood pressure, fasting glucose, cholesterol, and physical activity) assessed and classified into poor, intermediate, and optimal. Depression was assessed using the Clinical Interview Schedule-Revised (CIS-R). Poisson regression models, adjusted for sociodemographic factors and alcohol consumption, were employed. Stratified analyses were performed for age and sex. RESULTS: We included 9214 participants (mean age 52 ± 9 years, 48.6% women). Overall depression incidence at 3.8-year follow-up was 1.5%. Intermediate and poor ICH significantly increased the risk rate (RR) of developing depression (2.48 [95%CI 1.06-5.78] and 3 [1.28-7.03], respectively) at a 3.8-year follow-up. Higher ICH scores decreased the rate of depression development (RR = 0.84 [0.73-0.96] per metric). Stratified analyses were significant for women and adults < 55 years old. CONCLUSIONS: Poor cardiovascular health tripled depression risk at follow-up in otherwise healthy adults. Ameliorating cardiovascular health might decrease depression risk development.

Peripheral cytokine and chemokine alterations in depression: a meta-analysis of 82 studies.

OBJECTIVE: To conduct a systematic review and meta-analysis of studies that measured cytokine and chemokine levels in individuals with major depressive disorder (MDD) compared to healthy controls (HCs). METHOD: The PubMed/MEDLINE, EMBASE, and PsycINFO databases were searched up until May 30, 2016. Effect sizes were estimated with random-effects models. RESULT: Eighty-two studies comprising 3212 participants with MDD and 2798 HCs met inclusion criteria. Peripheral levels of interleukin-6 (IL-6), tumor necrosis factor (TNF)-alpha, IL-10, the soluble IL-2 receptor, C-C chemokine ligand 2, IL-13, IL-18, IL-12, the IL-1 receptor antagonist, and the soluble TNF receptor 2 were elevated in patients with MDD compared to HCs, whereas interferon-gamma levels were lower in MDD (Hedge's g = -0.477, P = 0.043). Levels of IL-1ß, IL-2, IL-4, IL-8, the soluble IL-6 receptor (sIL-6R), IL-5, CCL-3, IL-17, and transforming growth factor-beta 1 were not significantly altered in individuals with MDD compared to HCs. Heterogeneity was large (I2 : 51.6-97.7%), and sources of heterogeneity were explored (e.g., age, smoking status, and body mass index). CONCLUSION: Our results further characterize a cytokine/chemokine profile associated with MDD. Future studies are warranted to further elucidate sources of heterogeneity, as well as biosignature cytokines secreted by other immune cells.

Caracterização de estirpes de Campylobacter coli isoladas de carcaças de ovinos e de efluentes de abatedouro do estado de São Paulo / Characterization of Campylobacter coli strains isolated from the carcasses of sheep and of wastewater of abattoir in the state of São Paulo

Doença bacteriana zoonótica, a campilobacteriose é responsável mundialmente por frequentes casos de gastroenterite humana. Campylobacter spp. apresenta fator de virulência associado à diarreia, denominado toxina citoletal distensiva (CDT), sendo codificado pelos genes do complexo cdt. Os objetivos do presente estudo foram: 1) isolar e identificar estirpes de Campylobacter spp. de 102 suabes de carcaças e 102 suabes retais de ovinos (Ovis aries) e de sete amostras de água dos efluentes, antes e depois do tratamento de desinfecção de abatedouro localizado no estado de São Paulo; e 2) detectar, pela técnica de multiplex-PCR, a presença do complexo de genes cdt. Foram isoladas e identificadas, por métodos fenotípicos e genotípicos, sete estirpes de Campylobacter coli provenientes de 4/102 (3,92%) das amostras de suabes retais, 1/102 (0,98%) de suabes de carcaças e 2/7 (28,5%) das águas dos efluentes. Dos isolados de suabes retais, em 2/7 (28,6%) estirpes foi detectada a presença dos genes cdt. Trata-se do primeiro relato de isolamento de estirpes de Campylobacter coli provenientes de abatedouro de ovinos e das estirpes portadoras do complexo de genes cdt nessa espécie animal no Brasil.

A zoonosis and bacterial disease, campylobacteriosis is responsible for frequent cases of human gastroenteritis worldwide. Campylobacter spp. presents the virulence factor called cytolethal distensive toxine (CDT), responsible for diarrhea and codified by the cdt gene. The aims of this study were: 1) to isolate and identify Campylobacter spp. strains from 102 carcass swabs and 102 rectal swabs of sheep (Ovis aries) and seven samples of wastewater, before and after the disinfection treatment, collected from the abattoir of the state of São Paulo; and 2) to detect the presence of cdt gene complex by Multiplex-PCR in strains of Campylobacter spp. Seven strains of Campylobacter coli were isolated and identified by phenotypic and genotypic methods: 4/102 (3.92%) from rectal swabs, 1/102 (0.98%) from carcass swabs and 2/7 (28.5%) from wastewater. From the rectal swab samples 2/7 (28.6%) strains were detected with the cdt gene. This is the first report on the isolation of Campylobacter coli from sheep abattoir, and of strains carrying the cdt gene complex in this animal species in Brazil.

The Adverse Effects of Smoking on Health Outcomes in Bipolar Disorder: A Review and Synthesis of Biological Mechanisms.

Several biological systems are implicated in the neuroprogression of bipolar disorder including but not limited to cytokine levels, oxidative stress markers, monoamine levels, tryptophan catabolite and glutamate-mediated excitotoxicity, microglial activation as well as structural and functional changes. The high rate of smoking behaviour in individuals with bipolar disorder provides the impetus for exploring shared and discrete pathogenetic mechanisms. In addition to contributing to increased mortality, smoking activates several neurobiological effector systems implicated in the progression of bipolar disorder. Here, a narrative review provides evidence and putative mechanisms of comorbid effects of BD, cigarette use, and nicotine dependence, and discusses the clinical implications of these interactions.

Depression and Disturbed Bone Metabolism: A Narrative Review of the Epidemiological Findings and Postulated Mechanisms.

Major depressive disorder (MDD) is a pervasive chronic condition that contributes substantially to the global burden of disease and disability. Adding to the complexity of this disorder are numerous associated medical comorbidities with a bidirectional impact on morbidity and mortality. In recent years, osteoporosis has been increasingly identified as a significant comorbidity of MDD. This narrative review examines the literature to summarize key epidemiological studies and discuss postulated mechanisms of interaction. Epidemiological studies have repeatedly shown an increased co-prevalence of fractures and decreased bone mineral density (BMD) in MDD. The pathophysiological mechanism underlying this interaction is undoubtedly complex and multifactorial, and proposed pathways have varying levels of evidence from preclinical and clinical models. Conceptually, the mechanisms by which depression might influence bone metabolism can be categorized into biological, behavioral, iatrogenic, and comorbidity-related factors. Biological factors include the inflammatory-mood pathway, hypothalamic-pituitary-adrenal (HPA) axis dysregulation, metabolic dysfunction, and serotonin's direct and indirect effects on bone cells. Behavioral factors incorporate lifestyle choices typical in depressed patients, such as increased tobacco use or limited exercise. The prominent iatrogenic factor is the independent effects of anti-depressants on bone metabolism. Psychiatric and medical comorbidities common to both osteoporosis and MDD are also important to consider. Physical activity promotion, vitamin D supplementation, and routine BMD screening of MDD patients are simple interventions that might lead to improved outcomes for both conditions. An improved understanding of the underlying mechanisms may yield insights into novel prevention and treatment strategies to target osteoporosis and fractures in the MDD population.

Fibromyalgia and Bipolar Disorder: Emerging Epidemiological Associations and Shared Pathophysiology.

Fibromyalgia (FM) is a prevalent disorder defined by the presence of chronic widespread pain in association with fatigue, sleep disturbances and cognitive dysfunction. Recent studies indicate that bipolar spectrum disorders frequently co-occur in individuals with FM. Furthermore, shared pathophysiological mechanisms anticipate remarkable phenomenological similarities between FM and BD. A comprehensive search of the English literature was carried out in the Pubmed/MEDLINE database through May 10th, 2015 to identify unique references pertaining to the epidemiology and shared pathophysiology between FM and bipolar disorder (BD). Overlapping neural circuits may underpin parallel clinical manifestations of both disorders. Fibromyalgia and BD are both characterized by functional abnormalities in the hypothalamic-pituitary-adrenal axis, higher levels of inflammatory mediators, oxidative and nitrosative stress as well as mitochondrial dysfunction. An over-activation of the kynurenine pathway in both illnesses drives tryptophan away from the production of serotonin and melatonin, leading to affective symptoms, circadian rhythm disturbances and abnormalities in pain processing. In addition, both disorders are associated with impaired neuroplasticity (e.g., altered brain-derived neurotrophic factor signaling). The recognition of the symptomatic and pathophysiological overlapping between FM and bipolar spectrum disorders has relevant etiological, clinical and therapeutic implications that deserve future research consideration.

Isolation and characterization of mesenchymal stem cells from the yolk sacs of bovine embryos.

The yolk sac (YS) represents a promising source of stem cells for research because of the hematopoietic and mesenchymal cell niches that are present in this structure during the development of the embryo. In this study, we report on the isolation and characterization of YS tissue and mesenchymal stem cells (MSCs) derived from bovine YSs. Our results show that the YS is macroscopically located in the exocoelomic cavity in the ventral portion of the embryo and consists of a transparent membrane formed by a central sac-like portion and two ventrally elongated projections. Immunohistochemistry analyses were positive for OCT4, CD90, CD105, and CD44 markers in the YS of both gestational age groups. The MSCs of bovine YS were isolated using enzymatic digestion and were grown in vitro for at least 11 passages to verify their capacity to proliferate. These cells were also subjected to immunophenotypic characterization that revealed the presence of CD90, CD105, and CD79 and the absence of CD45, CD44, and CD79, which are positive and negative markers of MSCs, respectively. To prove their multipotency, the cells were induced to differentiate into three cell types, chondrocytes, osteoblasts, and adipocytes, which were stained with tissue-specific dyes (chondrogenic: Alcian Blue, osteogenic: Alizarin Red, and adipogenic: Oil Red O) to confirm differentiation. Gene expression analyses showed no differences in the patterns of gene expression between the groups or passages tested, with the exception of the expression of SOX2, which was slightly different in the G1P3 group compared to the other groups. Our results suggest that YS tissue from bovines can be used as a source of MSCs, which makes YS tissue-derived cells an interesting option for cell therapy and regenerative medicine.

Aspectos morfológicos macro e microscópicos do estômago de tamanduá-mirim (Tamandua tetradactyla) / Macroscopic and microscopic morphological aspects of lesser anteater stomach (Tamandua tetradactyla)

Diante do escasso conhecimento morfofisiológico do trato digestório do Tamandua tetradactyla, este trabalho pretendeu esclarecer a morfologia do estômago dessa espécie. Utilizaram-se seis espécimes, sendo três machos e três fêmeas jovens, provenientes da área de Mina Bauxita Paragominas - PA, doados, após morte por atropelamento, ao Laboratório de Pesquisa Morfológica Animal (LaPMA). Os animais foram fixados com solução aquosa de formaldeído (10 por cento), seguido de dissecação, avaliação da topografia do estômago, com posterior descrição, mensuração e coleta de material para microscopia. As análises topográfica e macroscópica revelaram que o estômago, unicavitário, com presença de pequena e grande curvaturas, estava localizado no antímero esquerdo da cavidade abdominal, ligado cranialmente ao esôfago e caudalmente ao duodeno, e apresentou um aumento de superfície na região pilórica (toro pilórico). Histologicamente, era constituído por epitélio colunar simples (região glandular), epitélio estratificado pavimentoso (região aglandular), lâmina própria, muscular da mucosa, submucosa, muscular circular interna, muscular longitudinal externa e serosa. A região do piloro apresentou um grande espessamento da camada muscular da mucosa, e foram observadas nesta região fossetas gástricas profundas. O estômago de T. tetradactyla revelou características morfológicas macroscópicas e histológicas semelhantes às espécies domésticas e selvagens...

Given the limited morphophysiological knowledge about the digestive tract of Tamandua tetradactyla, this study aimed to clarify the morphology of the stomach of this species. We used six young specimens, three males and three females, from the area of Paragominas bauxite mine - PA, donated after death by trampling, to the Research Laboratory Animal Morphological (LaPMA). The animals were fixed with an aqueous solution of formaldehyde (10 percent), followed by dissection, evaluation of the topography of the stomach, with further description, measurement and collection of material for microscopy. The topographical and macroscopic analysis showed that the stomach was located in left on-timer of the abdominal cavity, connected entirely to the esophagus and caudally to the duodenum, was unicavitary with the presence of small and large curvature and showed a surface increase in the pyloric region (torus pylorus). Histologically it was composed of simple columnar epithelium (glandular region), stratified epithelium (aglandular region), lamina propria, muscular of mucous, submucous, muscular internal circular, muscular external longitudinal and serosa. The region of the pylorus presented a great thickening of the muscular layer of mucous, and deep gastric pits were observed in this region. The stomach of T. tetradactyla revealed macroscopic and histological morphological characteristics similar to domestic and wild species...

Avaliação da atividade antibacteriana de extratos etanólico e de ciclohexano a partir das flores de camomila (Matricaria chamomilla L. ) / Evaluation of the antibacterial activity of ethanolic and cyclohexane extracts of chamomile flowers (Matricaria chamomilla L. )

Devido à crescente seleção de microrganismos resistentes aos antimicrobianos atuais, tem-se valorizado a busca por alternativas naturais. O presente estudo teve por objetivo avaliar a atividade antibacteriana de extratos etanólico e de ciclohexano de flores de camomila, espécie vegetal de uso antigo pela medicina tradicional, frente às bactérias ATCC Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli e Salmonella enterica subsp. enterica sorovar Typhimurium pelas técnicas de difusão em ágar e diluição em caldo. Foi observada inibição do crescimento de P. aeruginosa frente ao extrato etanólico bruto (1g/mL) na técnica de diluição em caldo, e confirmada pela técnica de difusão em ágar (halo de inibição de 10 mm de diâmetro). Para as demais bactérias testadas, os extratos e suas diluições não apresentaram efeito bacteriostático em nenhuma das técnicas. Pode-se concluir que o extrato etanólico bruto da camomila apresentou atividade antibacteriana frente à P. aeruginosa, porém não foi eficaz frente à S. aureus, E. coli e Salmonella enterica subsp. enterica sorovar Typhimurium. Portanto, são necessários novos estudos com diferentes linhagens de microrganismos, com o intuito de corroborar e assegurar os resultados apresentados, para definir o potencial antimicrobiano do extrato da camomila.

Due to the growing selection of microorganisms resistant to antimicrobial, the search for natural alternatives has become popular. The objective of the present study was to evaluate the antibacterial activity of ethanolic and cyclohexane extracts of chamomile flowers, a plant species long used by traditional medicine, against ATCC bacteria Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Salmonella enterica subsp. enterica serovar Typhimurium by agar diffusion and broth dilution techniques. Growth of P. aeruginosa was inhibited when crude ethanolic extract (1g/mL) was used broth dilution, and was confirmed by agar diffusion, (10 mm diameter inhibition zone). For the other bacteria tested, the extracts and their dilutions did not show any bacteriostatic effect in any of the techniques. It may be concluded that pure ethanolic extract of chamomile presents antibacterial action against P. aeruginosa, and none against S. aureus, E. coli e Salmonella enterica subsp. enterica sorovar Typhimurium. However, other studies with different strains of microorganisms may be useful in order to corroborate and ensure these results, to define evaluation of the antimicrobial activity of chamomile extract.

Antimicrobial activity of the Nisin Z producer Lactococcus lactis subsp. lactis Lc08 against Listeria monocytogenes in skim milk / Atividade antimicrobiana de Lactococcus lactis subsp. lactics Lc08 produtor de Nisina Z contra Listeria monocytogenes em leite desnatado

The presented study aimed to verify the effect of different pH values, enzyme solutions and heat treatments on the antimicrobial activity of the bacteriocinogenic strain Lactococcus lactis subsp. lactis Lc08 and to test their antimicrobial activity against Listeria monocytogenes in reconstituted skim milk at refrigeration temperatures. This strain was previously described as a nisin Z producer and capable of inhibiting L. monocytogenes growth in in vitro tests. The antimicrobial activity of the bacteriocin cell-free supernatant of Lc08 was sensitive to enzyme treatments (except papain). The pH values and heating (65ºC for 30min, 75ºC for 15s) had no apparent effect on the antimicrobial activity of the bacteriocin produced by Lc08. Only treatment at autoclave conditions result in loss of their antimicrobial activity. Lc08 presented antimicrobial activity against L. monocytogenes in the milk system after 12h at 25ºC. No effect was found at 7ºC. The results show the application viability of the Lc08 in food systems as a biopreservative against L. monocytogenes.

O presente estudo teve como objetivo verificar o efeito de diferentes valores de pH, soluções enzimáticas e tratamentos térmicos na atividade antimicrobiana da cepa bacteriocinogênica Lactococcus lactis subsp. lactis Lc08 e testar sua atividade antagonista contra Listeria monocytogenes em leite desnatado reconstituído em diferentes temperaturas de estocagem. Essa cepa já foi descrita como produtora de nisina Z e capaz de inibir o desenvolvimento de L. monocytogenes em testes in vitro. A atividade antimicrobiana do sobrenadante de Lc08 contendo a bacteriocina produzida e livre de células foi sensível ao tratamento pelas enzimas testadas (exceto papaína). A aplicação de diferentes valores de pH e o tratamento térmico (65ºC por 30 min, 75ºC por 15s) não influenciaram na atividade antimicrobiana da bacteriocina produzida por Lc08. Apenas o tratamento em autoclave resultou em perda da sua capacidade em inibir o desenvolvimento de L. monocytogenes. A cepa Lc08 apresentou atividade antagonista contra L. monocytogenes em leite após período de estocagem de 12h a 25ºC. Não foi observado efeito a 7ºC. Os resultados mostram a viabilidade de aplicação da cultura Lc08 ou de sua bacteriocina em produtos lácteos como bioconservador contra L. monocytogenes.

A spreadsheet for calculation of ingredients for standardization of requeijão cremoso / Planilha eletrônica para cálculos de ingredientes visando à padronização de requeijão cremoso

The standardization of requeijão cheese quality characteristics is a processing stage which is directly influenced by the chemical composition of the raw material. Fat and water concentrations are the main factors that cause variation in quality properties of the requeijão and, when not thoroughly controlled, may cause a lack in standardization of the food. Calculations of ingredients for effective standardization must be made in relation to the final product through of iterative calculations, which makes the manual process unfeasible. Thus, the objective of the present study was to develop a spreadsheet for ingredient calculation, with restrictions being the different concentrations of fat and water in order to standardize the final products. The spreadsheet was developed using the Excel 2007® Solver function, with just four input data: quantity of mass, water and fat content of the mass and fat content of the cream. The optimal solution was that which maximizes the amount of final product while meeting the stipulated constraints of fat and water. Results obtained with the spreadsheet were validated by processing the requeijão cheeses and comparison of the theoretical and practical parameters using the t-test. There was no significant difference between fat and water contents (p>0.10) estimated and verified in practice. Therefore, the use of the spreadsheet proved to be efficient for standardization of requeijão formulations.

A padronização das características de qualidade do requeijão é uma etapa do processamento, a qual é diretamente influenciada pela composição química da matéria-prima. Os teores de gordura e água são os principais fatores que acarretam variação nas propriedades de qualidade do requeijão e, quando não controlados minuciosamente, causam despadronização do alimento. Os cálculos de ingredientes para efetiva padronização devem ser realizados em relação ao produto final, por meio de cálculos iterativos, o que inviabiliza o processo manual. Nesse contexto, o presente trabalho teve por objetivo desenvolver uma planilha eletrônica para cálculo de ingredientes, tendo como restrições diferentes teores de gordura e água, visando à padronização dos produtos. A planilha foi desenvolvida utilizando-se o suplemento Solver do Excel2007®, sendo que os únicos parâmetros de entrada após a implementação da planilha foram: quantidade de massa, teor de gordura e de água na massa e teor de gordura do creme. A solução ótima foi a maximização da quantidade do produto final que satisfizesse as restrições dos teores de gordura e água estipulados. Os resultados obtidos pela planilha foram validados por meio do processamento dos requeijões e da comparação dos parâmetros teóricos e práticos utilizando-se o teste t. Não houve diferença significativa entre os teores de gordura e de água (p>0,10) estimados e verificados na prática. Portanto, a utilização da planilha eletrônica se apresenta eficiente para padronização de formulações de requeijão cremoso.

Detecção de fatores de virulência em estirpes de Campylobacter spp. isoladas de carcaças de suínos abatidos em frigoríficos / Virulence factors in strains of Campylobacter spp. isolated from the carcasses of swine slaughtered in abattoirs

Isolaram-se estirpes de Campylobacter spp. em amostras de carcaças (n=65), fezes (n=65) e linfonodos mesentéricos (n=65) de suínos abatidos em frigoríficos do estado de São Paulo e detectaram, pela técnica da Multiplex-PCR, a presença do complexo de genes cdt, responsáveis pela expressão do fator de virulência da toxina CDT. Do total de 195 amostras de origem suína, Campylobacter spp. foi isolado de 31 (15,9%), sendo 29 (93,6%) de amostras de suabe retal, 1/65 (3,2%) de suabe de carcaça e um (3,2%) de linfonodo. Vinte e oito estirpes de C. coli foram positivas para a detecção dos genes cdt, e três estirpes de C. jejuni foram negativas para a detecção desses genes. Foi detectada, pela primeira vez no estado de São Paulo, a presença dos genes cdt em 100% das estirpes de Campylobacter coli provenientes de suínos abatidos em frigoríficos.

The purposes of this study were to isolate and identify Campylobacter spp. strains from the carcasses (n=65), feces (n=65) and mesenteric lymph nodes (n=65) of swine slaughtered in abattoirs in the State of Sao Paulo and to detect the presence of the cdt gene complex - responsible for the expression of the virulence factor cytolethal distensive toxin - in these Campylobacter spp. strains through Multiplex-PCR. From 195 samples analyzed, Campylobacter spp. was isolated in 31 (15.9%): 29 (93,6%) samples of rectal swab, 1 (3.2%) carcass swab and 1 (3.2%) lymph node sample. The 28 strains of isolated C. coli were positive for CDT toxin genes and the three strains of isolated C. jejuni were negative for these genes. It was also the first time that the cdt gene cluster was detected in strains isolated from swine in the state of São Paulo. These findings indicate swine as a potential spreading source of virulent strains of Campylobacter coli, either for slaughterhouse staff or consumers of carcasses and sub products.

Caracterização molecular e fenotípica de estirpes de Escherichia coli produtoras de shiga-toxina (STEC) não-O157 de fezes e carcaças bovinas / Molecular and phenotypic characterization of shiga toxin producing Escherichia coli (STEC) non-O157 strains from bovine feces and carcass

Foram coletados 100 suabes retais e 100 suabes de carcaças bovinas em matadouros do estado de São Paulo, e um total de 326 estirpes de E. coli foram identificadas, sendo 163 de amostras retais e 163 de amostras de carcaça. Todos os isolados submetidos à PCR para detecção dos genes das toxinas Stx1 e Stx2 foram identificados como não-O157 e fenotipados pelo teste da citotoxicidade em células Vero. Das 26 estirpes que apresentaram apenas o gene stx1, das 56 que apresentaram apenas o gene stx2 e das 30 estirpes que apresentaram ambos os genes, 17 (65,4%), 42 (75%) e 22 (73,3%), respectivamente, foram positivas ao teste de citotoxicidade. Não houve diferença estatística entre os três perfis genéticos e na positividade ao teste de citotoxicidade. Os resultados mostram a alta frequência de expressão dos fatores de virulência das STEC de bovinos.

In the present study 100 rectal and 100 carcass swabs were collected from bovines at slaughterhouses in São Paulo state, and the total of 326 E. coli strains were identified (163 from rectal samples and 163 from carcass samples). All the isolates were submitted to PCR for Stx1 and Stx2 toxin gene detection and all strains were identified as non-O157 and phenotyped by the citotoxicity test in Vero cells. Out of 26 strains that presented only the stx1 gene, 56 that presented only the stx2 gene and 30 that presented both genes, 17 (65.4%), 42 (75%) and 22 (73.3%), respectively, were positive for the citotoxicity test. There was no statistically significant difference among these three toxinotyping profiles and positivity in the citotoxicity test, but the results show high frequency of virulence factor expression of bovine.

Animal models of prenatal immune challenge and their contribution to the study of schizophrenia: a systematic review

Prenatal immune challenge (PIC) in pregnant rodents produces offspring with abnormalities in behavior, histology, and gene expression that are reminiscent of schizophrenia and autism. Based on this, the goal of this article was to review the main contributions of PIC models, especially the one using the viral-mimetic particle polyriboinosinic-polyribocytidylic acid (poly-I:C), to the understanding of the etiology, biological basis and treatment of schizophrenia. This systematic review consisted of a search of available web databases (PubMed, SciELO, LILACS, PsycINFO, and ISI Web of Knowledge) for original studies published in the last 10 years (May 2001 to October 2011) concerning animal models of PIC, focusing on those using poly-I:C. The results showed that the PIC model with poly-I:C is able to mimic the prodrome and both the positive and negative/cognitive dimensions of schizophrenia, depending on the specific gestation time window of the immune challenge. The model resembles the neurobiology and etiology of schizophrenia and has good predictive value. In conclusion, this model is a robust tool for the identification of novel molecular targets during prenatal life, adolescence and adulthood that might contribute to the development of preventive and/or treatment strategies (targeting specific symptoms, i.e., positive or negative/cognitive) for this devastating mental disorder, also presenting biosafety as compared to viral infection models. One limitation of this model is the incapacity to model the full spectrum of immune responses normally induced by viral exposure.

Validação de nova proposta de espermocultura quantitativa aplicada a sêmen industrializado de touros / Validation of a new proposal of quantitative sperm culture for industrialized bull semen

O Ministério da Agricultura, Pecuária e Abastecimento (MAPA) tem discutido a obrigatoriedade da realização de espermocultura em sêmen industrializado não somente para garantia da biossegurança mas também por falhas na fertilização in vitro pela contaminação dos ovócitos com bactérias ubiquitárias e oportunistas da microbiota prepucial. A técnica quantitativa de Pour Plate, preconizada pela Organização Internacional de Epizootias, é operacionalmente difícil e de custo elevado para a rotina em centrais de inseminação artificial (CIAs). Desse modo, avaliou-se a técnica de contagem de bactérias viáveis por superfície (CBVS) em UFC/mL de sêmen industrializado de touros de CIAs, comparando-a à técnica de Pour Plate, com intuito de validação. Foram empregadas palhetas de sêmen de touros do Projeto Hungria do MAPA. A partir da diluição seriada de 10-1 a 10-5 do sêmem, foram empregadas, paralelamente, as técnicas de Pour Plate e CBVS. A média ou mediana de UFC/mL obtida em cada diluição foi comparada entre as técnicas, e não foram observadas diferenças estatísticas entre as duas técnicas quanto à quantificação de bactérias em UFC/mL, sugerindo a possibilidade de substituição da técnica de Pour Plate pela de CBVS, com vantagens de praticidade e menor custo.

The Brazilian Ministry of Agriculture (MAPA) has discussed the mandatory culture of industrialized semen, both to ensure biosefety, and to prevent in vitro fertilization problems caused by oocyte contamination with ubiquitous and opportunistic bacteria from preputial microbiota. Pour plate, a quantitative technique recommended by the World Organization for Animal Health (OIE), is operationally difficult and costly for routine analysis in Artificial Insemination Centers (AICs). The objectives of this study were to evaluate and validate viable superficial bacteria counts (VSBC), in CFU/mL, compared with pour plate counts, in industrialized bull semen samples from AICs. Semen straws from Projeto Hungria - MAPA bulls were used. VSBC and pour plate were carried out in parallel in serial dilutions of the samples, from 10-1 to 10-5. CFU/mL means or medians recorded in each dilution and technique were compared, and no statistical differences were observed between the two techniques regarding the quantification of bacteria in CFU/mL, suggesting that it may be possible to replace pour plate for CBSV, a cheaper and more practical technique.

Genes involved in protein metabolism of the probiotic lactic acid bacterium Lactobacillus delbrueckii UFV H2b20.

A basic requirement for the prediction of the potential use of lactic acid bacteria (LAB) in the dairy industry is the identification of specific genes involved in flavour-forming pathways. The probiotic Lactobacillus delbrueckii UFV H2b20 was submitted to a genetic characterisation and phylogenetic analysis of genes involved in protein catabolism. Eight genes belonging to this system were identified, which possess a closely phylogenetic relationship to NCFM strains representative, as it was demonstrated for oppC and oppBII, encoding oligopeptide transport system components. PepC, PepN, and PepX might be essential for growth of LAB, probiotic or not, since the correspondent genes are always present, including in L. delbrueckii UFV H2b20 genome. For pepX gene, a probable link between carbohydrate catabolism and PepX expression may exists, where it is regulated by PepR1/CcpA-like, a common feature between Lactobacillus strains and also in L. delbrueckii UFV H2b20. The well conserved evolutionary history of the ilvE gene is evidence that the pathways leading to branched-chain amino acid degradation, such as isoleucine and valine, are similar among L. delbrueckii subsp. bulgaricus strains and L. delbrueckii UFV H2b20. Thus, the involvement of succinate in flavour formation can be attributed to IlvE activity. The presence of aminopeptidase G in L. delbrueckii UFV H2b20 genome, which is absent in several strains, might improve the proteolytic activity and effectiveness. The nucleotide sequence encoding PepG revealed that it is a cysteine endopeptidase, belonging to Peptidase C1 superfamily; sequence analysis showed 99% identity with L. delbrueckii subsp. bulgaricus ATCC 11842 pepG, whereas protein sequence analysis revealed 100% similarity with PepG from the same organism. The present study proposes a schematic model to explain how the proteolytic system of the probiotic L. delbrueckii UFV H2b20 works, based on the components identified so far.

Amygdalar peptidergic circuits regulating noradrenergic locus coeruleus neurons: linking limbic and arousal centers.

The endogenous opioid peptides, met- or leu-enkephalin, and corticotropin-releasing factor (CRF) regulate noradrenergic neurons in the locus coeruleus (LC) in a convergent manner via projections from distinct brain areas. In contrast, the opioid peptide dynorphin (DYN) has been shown to serve as a co-transmitter with CRF in afferents to the LC. To further define anatomical substrates targeting noradrenergic neurons by DYN afferents originating from limbic sources, anterograde tract-tracing of biotinylated dextran amine (BDA) from the central amygdaloid complex was combined with immunocytochemical detection of DYN and tyrosine hydroxylase (TH) in the same section of tissue. Triple labeling immunocytochemistry was combined with electron microscopy in the LC where BDA was identified using an immunoperoxidase marker, and DYN and TH were distinguished by the use of sequential immunogold labeling and silver enhancement to produce different sized gold particles. Results show direct evidence of a monosynaptic pathway linking amygdalar DYN afferents with LC neurons. To determine whether DYN-containing amygdalar LC-projecting neurons colocalize CRF, retrograde tract-tracing using fluorescent latex microspheres injected into the LC was combined with immunocytochemical detection of DYN and CRF in single sections in the central amygdala. Retrogradely labeled neurons from the LC were distributed throughout the rostro-caudal extent of the central nucleus of the amygdala (CeA) as previously described. Cell counts showed that approximately 42% of LC-projecting neurons in the CeA contained both DYN and CRF. Taken with our previous studies showing monosynaptic projections from amygdalar CRF neurons to noradrenergic LC cells, the present study extends this by showing that DYN and CRF are co-transmitters in monosynaptic projections to the LC and are poised to coordinately impact LC neuronal activity.

Detecção dos genes da toxina citoletal distensiva em estirpes de Campylobacter jejuni isoladas de carcaças de frangos / Detection of cytolethal distending toxin genes in strains of Campylobacter jejuni isolated from broiler carcasses

Foram analisadas 80 amostras de sobrecoxas de frangos de corte resfriados provenientes de feiras livres e hipermercados do município de São Paulo, SP. Treze estirpes de Campylobacter spp. foram isoladas em 10 (12,5 por cento) sobrecoxas, sendo cinco amostras originárias de feiras livres e cinco de hipermercados. Onze estirpes foram identificadas como Campylobacter jejuni e duas como Campylobacter coli. As 11 estirpes foram confirmadas como C. jejuni pela PCR do gene da hipuricase (hip), e destas, quatro (36,4 por cento) apresentaram os três genes (cdtA, cdtB e cdtC) codificantes da toxina citoletal distensiva pela multiplex-PCR, sendo três estirpes provenientes de hipermercados e uma de feira livre. Observou-se a presença de estirpes virulentas de C. jejuni, portadoras do complexo de genes cdt, nas amostras de frango resfriado, não só na linha de abate, mas até o ponto final da cadeia de distribuição, nos dois principais centros de venda a varejo.

Eighty samples of refrigerated broiler thighs purchased in street markets and supermarkets in the city of São Paulo, SP, were analyzed. Thirteen Campylobacter spp. strains were isolated in 10 (12.5 percent) thighs, five of them from street market samples and other five from supermarkets. Eleven strains were identified as Campylobacter jejuni and two of them as Campylobacter coli. The 11 strains were confirmed to be C. jejuni using PCR for hippuricase (hip) gene. From these, multiplex-PCR showed that four (36.4 percent) strains presented the three genes (cdtA, cdtB, and cdtC) encoding cytolethal distending toxin: three strains from supermarket and one from street market samples. These results are important, because they demonstrate the presence of virulent C. jejuni strains in refrigerated broiler thigh samples, not only in the slaughterhouse but in the final point of the distribution chain, at the two most important food retail commercer.

Transplacental transfer of iron in the water buffalo (Bubalus bubalis): uteroferrin and erythrophagocytosis.

The objectives of this investigation were to understand transplacental transport of iron by secreted uteroferrin (UF) and haemophagous areas of water buffalo placenta and clarify the role(s) of blood extravasation at the placental-maternal interface. Placentomes and interplacentomal region of 51 placentae at various stages of gestation were fixed, processed for light and transmission electron microscopy, histochemistry and immunohistochemistry. Haemophagous areas were present in placentomes collected between 4 and 10 months of pregnancy. Perl's reaction for ferric iron was negative in placentomes, but positive in endometrial glands. Positive staining for UF indicated areas in which it was being taken up by phagocytosis and/or fluid phase pinocytosis in areolae of the interplacentomal mesenchyme, with little staining in endometrial stroma. Imunohistochemistry detected UF in trophectoderm of haemophagous regions of placentomes and in other parts of the foetal villous tree, but the strongest immunostaining was in the epithelial cells and lumen of uterine glands. Ultrastructural analyses indicated that erythrophagocytosis was occurring and that erythrocytes were present inside cells of the chorion that also contained endocytic vesicles and caveolae. Results of this study indicate that both the haemophagous areas of placentomes and the areolae at the interface between chorion and endometrial glands are important sites for iron transfer from mother to foetal-placental tissues in buffalo throughout pregnancy.