The O-antigen of Salmonella enterica serotype Enteritidis PT4: a significant factor in gastrointestinal colonisation of young but not newly hatched chicks

The lipopolysaccharide of Salmonella and other Gram negative pathogenic species has been implicated as a major virulence determinant and in this study we report the role of LPS of S. Enteritidis in the colonisation and persistent gastrointestinal infection of young poultry. The gene encoding the unique O-antigen ligase, waaL, was mutated by insertional inactivation in a well characterised S. Enteritidis strain, S1400/94. The waaL mutant, designated PCP, produced rough colonies on agar medium, did not agglutinate O9 antiserum, did not produce an LPS ladder on silver stained gels and was serum sensitive. PCP and a nalidixic acid marked derivative of S1400/94 (S1400/94 Nal(r)) were used to orally challenge young chicks, separately and together in competitive index experiments. At post-mortem examination of 1-day-old chicks challenged S1400/94 Nal(r) and PCP separately there were no significant differences in the numbers of S1400/94 Nal(r) and PCP bacteria in tissues sampled on days 1, 2. and 5. By day 42 after challenge S1400/94 Nal(r) bacteria were recovered in significantly higher numbers than PCP from the caecal contents (P < 0.001). In competitive index studies in the 1-day-old chick PCP colonised, invaded and persisted in lower numbers than S1400/94 Nal(r). In 4-week-old chicks challenged separately, PCP bacteria were recovered from all tissues examined in significantly lower numbers than S1400/94 Nal(r). In competitive index experiments in 4-week-old chicks, PCP was not detected at any site and at any time point. Therefore, the O-antigen of S. Enteritidis plays art important role in poultry infections although this role is less important in the newly hatched chick. Crown Copyright (C) 2004 Published by Elsevier B.V. All rights reserved.

Variable and strain dependent colonisation of chickens by Escherichia coli O157

The prevalence of enterohaemorrhagic Escherichia coli (EHEC) O157 in poultry is considered minimal compared with other species, especially ruminants. However, deliberate inoculation studies have shown that poultry are readily and persistently infected by this organism but that the mechanism of colonisation is independent of intimin, a recognised factor in host-EHEC interactions in mammalian species, and may be dependent upon flagella. Few strains of EHEC O157 have been tested in poultry and here 1-day-old and 6-week-old chicks were inoculated with seven non-toxigenic E. coli O157 strains in separate experiments. Persistence was measured semi-quantitatively by bacteriological assessment of E. coli O157 cultured from cloacal swabs (shedding score). In the 1-day-old chick model that was monitored for 43 days, all seven strains established well after inoculation. In the 6-week-old chicken model, one strain established and gave consistently high shedding for the duration of the experiment (156 days). Whereas of the remaining six strains, two persisted for 113 days, two persisted for 43 days, one persisted for 22 days and one strain was never detected.

BREEDING BIOLOGY OF KELP GULLS ON THE BRAZILIAN COAST

We studied clutch size, hatching and fledging success, and time necessary for chick Kelp Gulls (Larus dominicanus) to leave the nest throughout two breeding seasons (2004 and 2005) on Guararitama Island, Sao Paulo, Brazil. We followed 93 nests in 2004 and 97 nests in 2005. The average (+/- SD) clutch size was 2.09 +/- 0.64 in 2004 and 1.93 +/- 0.59 in 2005. Hatching success was 74% in 2004 and 53% in 2005, and fledging success was 54% in 2004 and 58% in 2005. Chicks grew quickly, following the linear equation y(t) = 61g + 17.03g X age (in days), and began to fly at 40 days. Received 11 August 2008. Accepted 28 August 2009.

Connexin36, an Essential Element in the Rod Pathway, Is Highly Expressed in the Essentially Rodless Retina of Gallus gallus

Electrical coupling provided by connexins (Cx) in gap junctions (GJ) plays important roles in both the developing and the mature retina. In mammalian nocturnal species, Cx36 is an essential component in the rod pathway, the retinal circuit specialized for night, scotopic vision. Here, we report the expression of Cx36 in a species (Gallus gallus) that phylogenetic development endows with an essentially rodless retina. Cx36 gene is very highly expressed in comparison with other Cxs previously described in the adult retina, such as Cx43, Cx45, and Cx50. Moreover, real-time PCR, Western blot, and immunofluorescence all revealed that Cx36 expression massively increased over time during development. We thoroughly examined Cx36 in the inner and outer plexiform layers, where this protein was particularly abundant. Cx36 was observed mainly in the off sublamina of the inner plexiform layer rather than in the on sublamina previously described in the mammalian retina. In addition, Cx36 colocalized with specific cell markers, revealing the expression of this protein in distinct amacrine cells. To investigate further the involvement of Cx36 in visual processing, we examined its functional regulation in retinas from dark-adapted animals. Light deprivation markedly up-regulates Cx36 gene expression in the retina, resulting in an increased accumulation of the protein within and between cone synaptic terminals. In summary, the developmental regulation of Cx36 expression results in particular circuitry-related roles in the chick retina. Moreover, this study demonstrated that Cx36 onto- and phylogenesis in the vertebrate retina simultaneously exhibit similarities and particularities. J. Comp. Neurol. 512:651-663, 2009. (C) 2008 Wiley-Liss, Inc.

Differential response of AMPA and NMDA glutamate receptors of Purkinje cells to aging of the chicken cerebellum

Aging can lead to cognitive, affective, learning, memory and motor deficits. Since the cerebellum and glutamatergic neurotransmission are involved in several of those functions, the present work aimed at studying the expression of AMPA and NMDA glutamate receptor subunits in the chick cerebellum during aging. Young (30 days old) and aged (ca. 4 years old) chickens (Gallus gallus) were used in order to evaluate the expression of GluR1, GluR2/3 and NR1 subunits. The cerebella of young and aged chickens were subjected to immunohistochemical and immunoblotting techniques. Numbers of GluR1, GluR2/3 and NR1-positive cells and optical density of the immunoblotting data were analyzed and submitted to statistical analysis using ANOVA and the Bonferroni post hoc test. Mean density of Purkinje cells stained for Giemsa, GluR1, GluR2/3 and NR1 in the cerebellum all showed a statistically significant decrease in aged animals when compared to the young animals (Giemsa, P < 0.01; GluRs and NR1, P < 0.03). However, the ratio of GluR1 and GluR2/3-positive Purkinje cells in relation the total number of Purkinje cells found in each time point decreased with aging (ca. 10%), whereas the ratio of NR1-positive cells increased (ca. 9%). The immunoblotting data showed a significant decrease of GluR1 (ca. 66%) and GluR2/3 (ca. 55%) protein expression with aging, but did not reveal changes for NR1. Our data suggest that aging can lead to differential changes in the pattern of expression of glutamate receptor subunits, which can underlie at least part of the cognitive and motor disorders found in aged animals. (c) 2010 Elsevier Ireland Ltd. All rights reserved.

Retinoic Acid and VEGF Delay Smooth Muscle Relative to Endothelial Differentiation to Coordinate Inner and Outer Coronary Vessel Wall Morphogenesis

Rationale: Major coronary vessels derive from the proepicardium, the cellular progenitor of the epicardium, coronary endothelium, and coronary smooth muscle cells (CoSMCs). CoSMCs are delayed in their differentiation relative to coronary endothelial cells (CoEs), such that CoSMCs mature only after CoEs have assembled into tubes. The mechanisms underlying this sequential CoE/CoSMC differentiation are unknown. Retinoic acid (RA) is crucial for vascular development and the main RA-synthesizing enzyme is progressively lost from epicardially derived cells as they differentiate into blood vessel types. In parallel, myocardial vascular endothelial growth factor (VEGF) expression also decreases along coronary vessel muscularization. Objective: We hypothesized that RA and VEGF act coordinately as physiological brakes to CoSMC differentiation. Methods and Results: In vitro assays (proepicardial cultures, cocultures, and RALDH2 [retinaldehyde dehydrogenase-2]/VEGF adenoviral overexpression) and in vivo inhibition of RA synthesis show that RA and VEGF act as repressors of CoSMC differentiation, whereas VEGF biases epicardially derived cell differentiation toward the endothelial phenotype. Conclusion: Experiments support a model in which early high levels of RA and VEGF prevent CoSMC differentiation from epicardially derived cells before RA and VEGF levels decline as an extensive endothelial network is established. We suggest this physiological delay guarantees the formation of a complex, hierarchical, tree of coronary vessels. (Circ Res. 2010;107:204-216.)

Rho Signaling Pathway and Apical Constriction in the Early Lens Placode

Epithelial invagination in many model systems is driven by apical cell constriction, mediated by actin and myosin II contraction regulated by GTPase activity. Here we investigate apical constriction during chick lens placode invagination. Inhibition of actin polymerization and myosin II activity by cytochalasin D or blebbistatin prevents lens invagination. To further verify if lens placode invaginate through apical constriction, we analyzed the role of Rho-ROCK pathway. Rho GTPases expression at the apical portion of the lens placode occurs with the same dynamics as that of the cytoskeleton. Overexpression of the pan-Rho inhibitor C3 exotoxin abolished invagination and had a strong effect on apical myosin II enrichment and a mild effect on apical actin localization. In contrast, pharmacological inhibition of ROCK activity interfered significantly with apical enrichment of both actin and myosin. These results suggest that apical constriction in lens invagination involves ROCK but apical concentration of actin and myosin are regulated through different pathways upstream of ROCK. genesis 49: 368-379, 2011. (C) 2011 Wiley-Liss, Inc.

Women of Substance : The Aspect of Education, Career and Female Identity in Pride and Prejudice and Bridget Jones's Diary

Although two hundred years separate Jane Austen and Helen Fielding and, subsequently, also their portrayals of society, the similarities outweigh the differences. When juxtaposing Pride and Prejudice and Bridget Jones’s Diary in the light of feminism it is evident that both books provide clear examples of the prevailing situation of women in each time and place. The aspects of the study, which are especially important today, show both the development and some degree of stagnation of women’s rights and identities.

A influência da alimentação com diferentes níveis de vitamina E e selênio sobre a produção de imunoglobulina Y (IgY) no soro de poedeiras leves vacinadas contra Escherichia coli e encefalomielite aviária

Foram estudados os efeitos da suplementação de vitamina E (VE) e selênio (Se) sobre a imunidade de galinhas vacinadas contra Escherichia coli (EC) patogênica para suínos e com o vírus da encefalomielite aviária (VEA). Foram avaliados peso corporal (PC), peso de ovos (PO) e produção de anticorpos (AcP) em noventa poedeiras (H&N nick chick) com 49 semanas de idade alimentadas à vontade com dietas suplementadas com 0, 50, 150, 250 e 250 UI VE/kg + 0,3 ppm de Se. Às 51 semanas de idade, metade das galinhas foram vacinadas contra EC e todas as aves foram vacinadas contra VEA. Duas semanas após, receberam uma segunda dose da vacina contra EC. Amostras de sangue foram coletadas semanalmente e a quantidade de IgY foi determinada por ELISA, como densidade ótica (DO). As aves vacinadas apresentaram maior DO do que as aves não vacinadas (P≤0,05). As DO foram significativamente aumentadas (P≤0,05) quando as aves vacinadas foram alimentadas com 50 e 150 UI de VE/kg e comparadas com as DO das aves que receberam a dieta de 250 UI de VE + Se. Se não afetou AcP, PC e PO. A vacinação e a VE suplementada não afetaram PC. O PO não foi afetado pela vacinação, mas foi maior (P≤0,05) nas semanas 3, 5 e 7, quando as poedeiras receberam 250 UI VE/kg e comparados com aquelas que receberam 50, 150 e 0. A AcP contra VEA foi influenciada pela VE (P≤0,13). Cento e cinqüenta UI VE e 0 UI VE aumentaram IgY produzida, quando comparados a 250 UI. Também o Se aumentou AcP. Os resultados deste estudo sugerem que níveis médios de VE aumentam a produção de IgY por poedeiras vacinadas e Se adicionado ao mais alto nível de VE também possui efeito imunomodulador.

Aspectos estruturais, farmacológicos e biológicos de fucanas da alga marrom sargassum vulgare

The present study examines the chemical composition and their effects on free radicals, inflammation, angiogenesis, coagulation, VEGF effects and cellular proliferation of a polysaccharides from alga Sargassum vulgare. The sulfated polysaccharide was extracted from brown seaweed by proteolysis with enzymes maxataze. The presence of proteins and sugars were observed in crude polysaccharides. Fractionation of this crude extract was made with growing concentration of acetone (0.3-1.5 v) and produced four groups of polysaccharides. Anionic polysaccharides from brown seaweed Sargassum vulgare, SV1and PSV1 were fractionated (SV1) and purified (PSV1), and displayed with high total sugars and sulfate content and very low level of protein. This fucan SV1 contains low levels of protein and high carbohydrate and sulfate content. This polysaccharides prolonged activated partial thromboplastin time (aPTT) at 50 μg (>240 s). SV1 was found to have no effect on prothrombin time (PT), corresponding to the extrinsic pathway of coagulation. SV1 exhibits high antithrombotic action in vivo, with a concentration ten times higher than heparin. Polysaccharides from S. vulgare promoted direct inhibition enzymatic activity of thrombin and stimulated enzymatic activity of FXa. SV1 showed optimal inhibitory activity of thrombin (50.2±0.28%) at a concentration of 25 μg/mL. Its antioxidant action on scavenging radicals by DPPH was (22%), indicating the polymer has no cytotoxic action (hemolytic) on ABO and Rh blood types in different erythrocyte groups and displays strong anti-inflammatory action on all concentrations tested in the carrageenan-induced paw edema model, demonstrated by reduced edema and cellular infiltration. Angiogenesis is a dynamic process of proliferation and differentiation. It requires endothelial proliferation, migration, and tube formation. In this context, endothelial cells are a preferred target for several studies and therapies. The antiangiogenic efficacy of polysaccharides was examined in vivo in the chick chorioallantoic membrane (CAM) model by using fertilized eggs. Decreases in the density of the capillaries were assessed and scored. The results showed that SV1 and PSV1 have an inhibitory effect on angiogenesis. These results were also confirmed by inhibition tubulogenesis in rabbit aorta endothelial cell (RAEC) in matrigel. These compounds were assessed in Apoptosis assay (Annexin V - FITC / PI) and cell viability by MTT assay of RAEC. These polysaccharides do not affect the viability and do not have apoptotic or necrotic action. RAEC cell when incubated with SV1 and PSV1showed inhibition of VEGF secretion, observed when compounds were incubated at 25, 50 and 100 μg/μL. The VEGF secretion with the RAEC cell line for 24 h, was more effective for PSV1 at 50 μg/μL(71.4%) than SV1 100 μg/μL (75.9%). SV1 and PSV1 had an antiproliferative action (47%) against tumor cell line HeLa. Our results indicate that these sulfated polysaccharides have antiangiogenic and antitumoral actions

Efeito do uso de probiótico sobre o desempenho e atividade de enzimas digestivas de frangos de corte

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

Yeast extract and prebiotic in pre-initial phase diet for broiler chickens raised under different temperatures

Objetivou-se nesta pesquisa avaliar o desempenho, o rendimento de carcaça e a morfometria intestinal de frangos de corte criados em diferentes temperaturas e que receberam na fase pré-inicial ração contendo ou não extrato de leveduras e prebiótico. Foram utilizados 1.440 pintos machos de 1 dia de idade, criados em diferentes câmaras climáticas. As rações, acrescidas ou não de extrato de leveduras e prebiótico, foram oferecidas somente na fase pré-inicial (1 a 7 dias). A partir do oitavo dia, todas as aves receberam a mesma ração, reajustada de acordo com as recomendações usuais. Adotou-se o delineamento experimental inteiramente casualizado em arranjo fatorial 3 × 2 × 2, composto de três temperaturas de criação (calor, conforto e frio) e dois níveis de extrato de leveduras (com ou sem) e prebiótico (com ou sem). O desempenho das aves foi avaliado considerando o ganho de peso, o consumo de ração, a conversão alimentar e a viabilidade aos 42 dias de idade. Também foram avaliados o rendimento de carcaça e a morfometria intestinal. O calor ambiente prejudicou o desempenho e o rendimento de carcaça. A inclusão de prebiótico na ração pré-inicial aumentou o ganho de peso e melhorou a conversão alimentar das aves criadas no calor. A inclusão dos produtos na ração de frangos de corte criados em ambiente de calor e no frio tem efeito benéfico sobre as vilosidades das aves.

Salmonella spp investigation in transport boxes of day-old birds

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

Efeito do processamento da soja integral sobre a energia metabolizável e a digestibilidade dos aminoácidos para aves

Essa pesquisa foi desenvolvida para se avaliar o valor nutricional da soja integral submetida a diferentes processamentos. Quatro ensaios de metabolismo foram conduzidos para determinar os coeficientes de digestibilidade da matéria seca (CDMS), do extrato etéreo (CDEE) e dos aminoácidos e os valores de energia metabolizável aparente e verdadeira da soja integral desativada (SID) e extrusada (SIE) e das misturas de farelo de soja com óleo degomado de soja (FSO) ou com óleo ácido de soja (FSOA). Nos ensaios um e dois, utilizou-se a metodologia tradicional de coleta total de excretas com pintos e galos, respectivamente. A metodologia da alimentação forçada com galos adultos intactos foi utilizada no ensaio três, e com galos cecotomizados, no ensaio quatro. Os CDMS e CDEE e a energia metabolizável determinados com galos foram superiores aos determinados com pintos. Os maiores valores de CDMS, CDEE e de energia metabolizável foram obtidos para FSO, seguidos pelos da SIE e FSOA, e os menores, para a SID. O processo de extrusão proporcionou melhores resultados no aproveitamento da gordura do grão de soja e, conseqüentemente, da energia. Entretanto, a digestibilidade dos aminoácidos não foi influenciada pelos processamentos. Os diferentes processamentos conferiram à soja integral características nutricionais que se distinguiram, principalmente quanto ao valor de energia metabolizável, que também variou com a idade das aves.

Desempenho, eficiência de utilização dos nutrientes e estrutura do trato digestório de pintos de corte alimentados na fase pré-inicial com rações de diferentes formas físicas

Este estudo foi realizado com o objetivo de avaliar o efeito da forma física da ração sobre o desempenho, a utilização dos nutrientes das rações e o desenvolvimento do trato disgestório de pintos de corte na primeira semana de vida. Foram utilizados 150 pintos machos Cobb distribuídos ao acaso em três tratamentos com cinco repetições de dez aves. Os tratamentos consistiram das formas físicas farelada, triturada e peletizada de uma ração pré-inicial comercial. Com exceção da moela, os órgãos do trato digestório e a estrutura morfométrica do intestino não foram influenciados pela forma física da ração. Maior digestibilidade do N foi obtida com a ração peletizada, enquanto a ração triturada possibilitou maior digestibilidade do EE. A energia metabolizável determinada para a ração farelada foi menor que a obtida nas demais formas físicas da ração. As aves alimentadas com ração farelada tiveram menor consumo de ração e ganho de peso e pior conversão alimentar. Ingeriram ainda menos energia, apresentaram menores retenções de proteína, gordura e energia corporal e utilizaram maior proporção da energia ingerida para produção de calor. As rações pré-iniciais na forma peletizada ou triturada possibilitam maior aproveitamento dos nutrientes da ração pelos pintos e melhor desempenho.