Nucleotides, β-glucans, ascorbic acid, α-tocoferol, and different concentrations of a vitamin-mineral premix promote growth of Nile tilapia juveniles / [Nucleotídeos, β-glucanos, ácido ascórbico, α-tocoferol e diferentes concentrações de uma pré-mistura de vitaminas e minerais promovem o crescimento de juvenis de tilápia do Nilo]

ABSTRACT Feed additives, such as β-glucans, nucleotides, ascorbic acid (vitamin C), and α-tocopherol (vitamin E) can improve fish immunity and contribute to enhanced zootechnical performance of the fish. This study aimed to evaluate the effect of different concentrations of vitamin- mineral premix with or without the inclusion of an immunostimulant boost (β-glucans, nucleotides and vitamins C and E) on the zootechnical performance, hemato-immunological parameters, histological changes, and survival of Nile tilapia juveniles. To this end, isocaloric and isoproteic diets were prepared with three different concentrations of vitamin-mineral premix (1.0, 1.5, and 2.0 kg ton-1); additionally, the effects of 4% immunostimulant boost in the diet were examined considering six diets (without the immunostimulant boost: 0.1%, 0.15%, and 0.2%, and with the immunostimulant boost: 0.1% I, 0.15% I, and 0.2% I). We used 24 experimental units, each including 15 fish. Nile tilapia juveniles (1.88 g±0.25) were fed for 50 d with supplemented diets. Further, zootechnical indexes; histological changes in the liver, spleen, and intestine; intestinal morphology; and hemato-immunological parameters were evaluated. The fish that received supplementation showed higher zootechnical values, compared to those that did not receive the supplementation of immunostimulant reinforcement. Weight gain (30.45g±3.06), daily weight gain (0.60g±0.06), final weight (32.41g±3.15) and feed conversion (0.95±0.06) were higher in tilapia fed with an inclusion concentration of 0.2%. Hematological parameters were not affected by either the pre-mix concentrations or the booster of the immunostimulant. There was an increase in the number of intestinal folds, length of the fold, total area and number of goblet cells in the group supplemented with immunostimulant reinforcement. Supplementation with the immunostimulant promoted growth and improved intestinal morphology and immunological parameters of Nile tilapia juveniles.

RESUMO Os aditivos alimentares, tais como β-glucanos, nucleotídeos, ácido ascórbico (vitamina C) e α-tocoferol (vitamina E), podem melhorar a imunidade dos peixes e contribuir para um melhor desempenho zootécnico. Este estudo teve como objetivo avaliar o efeito de diferentes concentrações de premix vitamínico mineral com ou sem a inclusão de um imunoestimulante (β-glucanos, nucleotídeos e vitaminas C e E) no desempenho zootécnico, nos parâmetros hematoimunológicos, nas alterações histológicas e na sobrevivência dos juvenis de tilápia-do-Nilo. Para esse fim, foram preparadas dietas isocalóricas e isoproteicas com três concentrações diferentes de premix vitamínico e mineral (1,0, 1,5, e 2,0kg ton-1); além disso, foram examinados os efeitos da inclusão de 4% de imunoestimulante na dieta, considerando-se seis dietas (sem o imunoestimulante: 0,1%, 0,15% e 0,2%, e com a inclusão de imunoestimulante: 0,1% I, 0,15% I e 0,2% I). Foram utilizadas 24 unidades experimentais, cada uma com 15 peixes. Os juvenis de tilápia-do-Nilo (1,88g±0,25) foram alimentados, durante 50 dias, com as dietas suplementadas. Além disso, foram avaliados índices zootécnicos; alterações histológicas no fígado, baço e intestino; morfologia intestinal; e parâmetros hematoimunológicos. Os peixes que receberam suplementação apresentaram valores zootécnicos mais elevados, em comparação aos que não receberam a suplementação de reforço imunoestimulante. O ganho de peso (30,45g±3,06), o ganho de peso diário (0,60g ± 0,06), o peso final (32,41g±3,15) e a conversão alimentar (0,95±0,06) foram maiores nas tilápias alimentadas com uma concentração de inclusão de 0,2%. Os parâmetros hematológicos não foram afetados nem pelas concentrações de premix nem pelo reforço do imunoestimulante. Foi observado o aumento no número de pregas intestinais, no comprimento da prega, na área total e no número de células caliciformes no grupo suplementado com o boost imunoestimulante. A suplementação com o imunoestimulante promoveu o crescimento e melhorou a morfologia intestinal e os parâmetros imunológicos de juvenis de tilápia-do-Nilo.

In vitro evaluation of anti-fungal activity of tropicamide against strains of Candida spp. resistant to fluconazole in planktonic and biofilm form.

Candida spp. is considered to be the third or fourth most common cause of bloodstream infections associated with healthcare services in the world. Currently, several strains exhibit resistance to the traditional treatments, making the development of new therapeutic molecules necessary. Drug repositioning is an alternative that can be used to work around problems such as toxicity, cost and time in the development of new drugs. This study aims to evaluate the in vitro antifungal effect of tropicamide, molecule of anticholinergic action, against planktonic cells of Candida spp. and biofilm of C. albicans. Six strains of different Candida species were used to determine the minimum inhibitory concentration (MIC) of tropicamide and fluconazole according to CLSI document M27-A3 and one strain of C. albicans was used to evaluate the activity of tropicamide against biofilms. In concentrations of 64µg/mL, the tropicamide exhibited 50% of inhibitory activity in planktonic cell and in concentrations of 128µg/mL is able to inhibit the formation of C. albicans biofilm. Despite the inhibitory activity shown at the present study, the use of a larger number of strains, as well as in vivo cytotoxicity assays, is necessary to confirm the hypothesis that tropicamide can be used as an adjuvant agent in the treatment of infections by the Candida genus.

Characterization of novel human intragenic antimicrobial peptides, incorporation and release studies from ureasil-polyether hybrid matrix.

Intragenic antimicrobial peptides (IAPs) are internal sequences of proteins with physicochemical similarities to Antimicrobial Peptides (AMPs) that, once identified and synthesized as individual entities, present antimicrobial activity. Many mature proteins encoded by the genomes of virtually any organism may be regarded as inner reservoirs of IAPs, conferring them ample biotechnological potential. However, IAPs may also share shortcomings with AMPs, such as low half-life in biological media and non-specific adsorption in eukaryotic cells. The present manuscript reports a translational approach that encompasses the uncovering of two novel IAPs from human proteins as well as the first results concerning the incorporation and sustained release of one of these peptides from ureasil-polyether hybrid polymeric films. For such, the software Kamal was used to scan putative IAPs in the human proteome, and two peptides, named Hs05 and Hs06, were identified, synthesized, and tested as antimicrobials. Biophysical assays were conducted using model phospholipid vesicles and 1H NMR solution structures in phospholipid micelles were obtained for the IAP Hs05. This peptide was incorporated in a polymeric matrix composed of the ureasil/PPO-PEO-PPO triblock copolymer, and the resulting films were evaluated by atomic force microscopy and imaging mass spectrometry. The release rate of Hs05 from the polymeric matrix was assessed and the antimicrobial activity of Hs05-loaded hybrid polymeric films was evaluated against the bacterium Escherichia coli. This study represents the first steps towards the development of polymeric films enriched with IAPs obtained from the human proteome as sustained release devices for topical application.

Histatin-5 induces the reversal of Pdr5p mediated fluconazole resistance in Saccharomyces cerevisae.

BACKGROUND: Candidiasis is a major opportunistic fungal infection in humans. The low number of antifungal drugs available to treat Candida infections and the increasing incidence of multidrug resistant (MDR) strains point to an urgent need of identifying new therapeutic options. The role of salivary components can provide insights for the development of new methodologies of control. OBJECTIVE: The aim of this study was to evaluate the ability of histatin-5, a constitutive immunological peptide present in saliva, in reversing fungal MDR phenotype, using a resistant Saccharomyces cerevisiae strain as model of study. RESULTS: A total of 2.5µg and 5µg of histatin-5 revealed to be able to chemosensitize (to revert antifungal resistance) a MDR strain to fluconazole impairing its intrinsic resistance. The presence of histatin-5 decreased the strain growth when associated to fluconazole, and also assisted in the retention of rhodamine 6G within cell cytoplasm. The ATPase activity of Pdr5p, an ABC efflux transporter, was significantly reduced up to 65% within physiological concentration of the peptide. CONCLUSION: Results revealed that histatin-5 is able to revert MDR phenotype and may be considered a potential alternative MDR inhibitor. Since Pdr5p is homologous to Candida albicans CaCdr1p and CaCdr2p, data obtained might be extrapolated to these transporters, inferring that associating fluconazole and histatin-5 may be a useful tool to circumvent failure treatments of infections caused by Candida MDR strains.

Effect of resistant starch on the intestinal health of old dogs: fermentation products and histological features of the intestinal mucosa.

The effects of resistant starch (RS) intake on nutrient digestibility, microbial fermentation products, faecal IgA, faecal pH, and histological features of the intestinal mucosa of old dogs were evaluated. The same formulation was extruded in two different conditions: one to obtain elevated starch cooking degree with low RS content (0.21%) and the other lower starch cooking with high RS content (1.46%). Eight geriatric Beagles (11.5 ± 0.38 years old) were fed each diet for 61 days in a crossover design. Food intake, nutrient digestibility, fermentation products, faecal pH, and faecal IgA were examined via variance analysis. Histological results of intestinal biopsies were assessed via Wilcoxon test for paired data. The morphometric characteristics of large intestine crypts were evaluated via paired t tests (p < .05). Protein, fat, and energy digestibilities were higher for the low-RS diet (p < .05). Dogs receiving the high-RS diet had lower faecal pH and higher values for propionate, butyrate, total volatile fatty acids, and lactate (p < .05). No differences between diets were found in the histological parameters of the gut mucosa, and only a tendency for deeper crypts in the descending colon was observed for dogs fed the high-RS diet (p = .083). The intake of a corn-based kibble diet manufactured with coarse ground raw material and low starch gelatinization to obtain 1.4% of RS affected microbial fermentation products and faecal pH and tended to increase crypt depth in the descending colon of old dogs.

Synthetic Organotellurium Compounds Sensitize Drug-Resistant Candida albicans Clinical Isolates to Fluconazole.

Invasive Candida albicans infections are a serious health threat for immunocompromised individuals. Fluconazole is most commonly used to treat these infections, but resistance due to the overexpression of multidrug efflux pumps is of grave concern. This study evaluated the ability of five synthetic organotellurium compounds to reverse the fluconazole resistance of C. albicans clinical isolates. Compounds 1 to 4, at <10 µg/ml, ameliorated the fluconazole resistance of Saccharomyces cerevisiae strains overexpressing the major C. albicans multidrug efflux pumps Cdr1p and Mdr1p, whereas compound 5 only sensitized Mdr1p-overexpressing strains to fluconazole. Compounds 1 to 4 also inhibited efflux of the fluorescent substrate rhodamine 6G and the ATPase activity of Cdr1p, whereas all five of compounds 1 to 5 inhibited Nile red efflux by Mdr1p. Interestingly, all five compounds demonstrated synergy with fluconazole against efflux pump-overexpressing fluconazole-resistant C. albicans clinical isolates, isolate 95-142 overexpressing CDR1 and CDR2, isolate 96-25 overexpressing MDR1 and ERG11, and isolate 12-99 overexpressing CDR1, CDR2, MDR1, and ERG11 Overall, organotellurium compounds 1 and 2 were the most promising fluconazole chemosensitizers of fluconazole-resistant C. albicans isolates. Our data suggest that these novel organotellurium compounds inhibit pump efflux by two very important and distinct families of fungal multidrug efflux pumps: the ATP-binding cassette transporter Cdr1p and the major facilitator superfamily transporter Mdr1p.

Tumor estromal gastrointestinal em cães: estudo clínico-anatomopatológico / Gastrointestinal stromal tumors in dogs: a clinicopathologic study

Os tumores estromais gastrointestinais (GIST) são incomuns e apresentam elevada similaridade histológica com as neoplasias de músculo liso e de nervo periférico. Este trabalho relata e caracteriza GIST em cães do ponto de vista epidemiológico, clínico-laboratorial, anatomopatológico e imuno-histoquímico, com base na análise retrospectiva de três cães, machos, com média de idade de 10 anos, variando de porte médio a grande e peso médio de 18.5±6kg. As alterações clínicas foram progressivas e inespecíficas de quadro gastrointestinal crônico, tais como prostração, anorexia, perda de peso e melena. Anemia não regenerativa e leucocitose foram os principais achados laboratoriais. Macroscopicamente, os tumores apresentavam média de 19cm no eixo maior, localizados em jejuno e ceco, com a superfície ulcerada, e exibiam áreas de necrose. Na avaliação microscópica, os casos apresentaram proliferação de células fusiformes, arranjadas em feixes longos densamente celulares, que foram vimentina e CD117 positivas, com diferenciação neural (S100 positivo), muscular (actina e desmina positivas) e anaplásica, cuja morfologia caracteriza neoplasias malignas com sobrevida de um dia a 12 meses. O GIST deve ser um dos diagnósticos diferenciais a serem considerados entre as neoplasias mesenquimais do trato gastrointestinal que acometem cães, e o diagnóstico final necessita de imuno-histoquímica.(AU)

Gastrointestinal stromal tumors (GIST) are uncommon and show high histological similarity with smooth muscle and peripheral neuron tumors. This paper reported three cases of gastrointestinal stromal tumor (GIST) showing its clinical and laboratorial presentation, and gross, microscopical and immunohistochemical features. Clinical signs and symptoms of the animals studied were progressive and characterized by nonspecific chronic gastrointestinal disorders such as depression, anorexia, weight loss and melena. Non regenerative anemia and leukocytosis were the main laboratory findings. Grossly, tumors were 19cm big, ulcerated and with necrotic areas. Microscopically, the tumors were malignant spindle cell proliferation with positivity to vimentine and CD117, with neural (S100+), myogenic (desmine + and smooth muscle actine+) and anaplasic differentiation. GIST should be one of the differential diagnoses to be considered among the mesenchymal neoplasms of the gastrointestinal tract of dogs.(AU)

Tumor estromal gastrointestinal em cães: estudo clínico-anatomopatológico / Gastrointestinal stromal tumors in dogs: a clinicopathologic study

Gastrointestinal stromal tumors (GIST) are uncommon and show high histological similarity with smooth muscle and peripheral neuron tumors. This paper reported three cases of gastrointestinal stromal tumor (GIST) showing its clinical and laboratorial presentation, and gross, microscopical and immunohistochemical features. Clinical signs and symptoms of the animals studied were progressive and characterized by nonspecific chronic gastrointestinal disorders such as depression, anorexia, weight loss and melena. Non regenerative anemia and leukocytosis were the main laboratory findings. Grossly, tumors were 19cm big, ulcerated and with necrotic areas. Microscopically, the tumors were malignant spindle cell proliferation with positivity to vimentine and CD117, with neural (S100+), myogenic (desmine + and smooth muscle actine+) and anaplasic differentiation. GIST should be one of the differential diagnoses to be considered among the mesenchymal neoplasms of the gastrointestinal tract of dogs.(AU)

Os tumores estromais gastrointestinais (GIST) são incomuns e apresentam elevada similaridade histológica com as neoplasias de músculo liso e de nervo periférico. Este trabalho relata e caracteriza GIST em cães do ponto de vista epidemiológico, clínico-laboratorial, anatomopatológico e imuno-histoquímico, com base na análise retrospectiva de três cães, machos, com média de idade de 10 anos, variando de porte médio a grande e peso médio de 18.5±6kg. As alterações clínicas foram progressivas e inespecíficas de quadro gastrointestinal crônico, tais como prostração, anorexia, perda de peso e melena. Anemia não regenerativa e leucocitose foram os principais achados laboratoriais. Macroscopicamente, os tumores apresentavam média de 19cm no eixo maior, localizados em jejuno e ceco, com a superfície ulcerada, e exibiam áreas de necrose. Na avaliação microscópica, os casos apresentaram proliferação de células fusiformes, arranjadas em feixes longos densamente celulares, que foram vimentina e CD117 positivas, com diferenciação neural (S100 positivo), muscular (actina e desmina positivas) e anaplásica, cuja morfologia caracteriza neoplasias malignas com sobrevida de um dia a 12 meses. O GIST deve ser um dos diagnósticos diferenciais a serem considerados entre as neoplasias mesenquimais do trato gastrointestinal que acometem cães, e o diagnóstico final necessita de imuno-histoquímica.(AU)

Molecular detection of chicken parvovirus in broilers with enteric disorders presenting curving of duodenal loop, pancreatic atrophy, and mesenteritis.

Enteric disorders are an important cause of economic losses in broiler chickens worldwide. Several agents have been associated with enteric problems, such as viruses, bacteria, and parasites. In this study, broiler chickens showing signs of enteric disorders were subjected to molecular diagnosis for several viral agents and also for pathological examination for elucidating this problem. Thus, the chickens were screened for avian nephritis virus (ANV), chicken astrovirus (CAstV), avian rotavirus (ArtV), avian reovirus (AReoV), infectious bronchitis virus (IBV), fowl adenovirus group I (FAdV-1), and chicken parvovirus (ChPV). Postmortem examinations revealed a curving of the duodenal loop (J-like appearance) and intestines filled with liquid and gaseous content. Histopathological analysis of the duodenal loop showed pancreatic atrophy, acute mesenteritis, and enteritis. PCR results showed that ChPV was the sole viral agent detected in samples with lesions such as the curved duodenal loop and pancreatic atrophy. Molecular characterization of the nucleotide and deduced amino acid sequences revealed a high similarity with other strains of ChPV from Brazil, Canada, United States, Europe, and Asia. These findings suggest an association between ChPV and the development of enteritis, pancreatitis, and pancreatic atrophy, which may lead to curling of the duodenal loop. Together, these alterations may disrupt the normal functioning of the digestive system, diminishing digestion and the absorption of dietary nutrients and consequently leading to reduced weight gain, flock impairment, dwarfism, and an elevated feed conversion rate.

Virulence and antimicrobial resistance of Klebsiella pneumoniae isolated from passerine and psittacine birds.

Klebsiella pneumoniae is considered one of the most important Gram-negative opportunistic pathogens. The contact between humans and birds poses health risks to both. The aim of this study was to investigate the resistance and virulence of K. pneumoniae isolates from psittacines and passerines, seized from illegal trade in Brazil. We analysed 32 strains isolated from birds of the orders Psittaciformes and Passeriformes by polymerase chain reaction (PCR) for virulence factor genes. Antibiotic resistance was assessed by disk diffusion assay and PCR. The results indicated that fimH (100%), uge (96.8%), kfu (81.2%) and irp-2 (68.7%) were the most common virulence genes, followed by kpn (46.8%), K2 (43.7%), mrkD (34.3%) and iroN (15.6%). The combination of virulence genes resulted in a great diversity of genotypes and the heterogeneity of the strains is also confirmed in the analysis by amplified fragment length polymorphism. The susceptibility profiles of the K. pneumoniae showed 25% of multiple antibiotic resistance strains. We identified seven strains that presented non-extended spectrum beta lactamase blaSHV variants SHV-1 and SHV-11 and one strain positive to the blaTEM-1 gene. Plasmid-mediated quinolone resistance was present in 10 strains (10/32). The data obtained in this study reveal the pathogenic potential of this pathogen and highlight the need for surveillance and monitoring.

Exchange of natural enemies for biological control: is it a rocky road?-the road in the Euro-Mediterranean region and the South American common market.

The access and benefit sharing (ABS) regulations from the Convention on Biological Diversity (CBD) for the use of natural resources became an important issue because the biodiversity of developing countries was heavily accessed and unilaterally exploited by pharmaceutical and seed companies. However, natural enemies used for biological control are living and unmodified genetic resources which cannot be patented and have been treated as resources such as drugs, seeds, or other commercial products. Consequently, the ABS requirements have limited not only the use of natural enemies but also the positive effects that scientifically supported biological control strategies have on the society, the environment, and the economy, reducing problems of pesticide residues, water and soil contamination, and non-target effects. During the last several years, the biological control scientific community has faced new and extremely complicated legislation dictated by a high and diverse number of governmental agencies at different levels, making the access to natural resources for biocontrol purposes a rocky road. Society at large should be aware of how the strict ABS regulations affect the use of natural enemies as biological resources to secure food production, food safety, and global environmental protection. We discuss in here the current difficulties derived from CBD for the exchange of natural enemies taking as example the Euro-Mediterranean region, Argentina, and Brazil to demonstrate how long and diverse are the steps to be followed to obtain the required permits for access and exportation/importation of natural enemies. We then argue that the public visibility of biocontrol strategies should be increased and their benefits highlighted in order to persuade legislators for the development of a less bureaucratic, more expedient, and more centralized regulatory frame, greatly favoring the practice and benefits of biological control. We finally propose a general framework in which ABS issues should be dealt in ways to attend the CBD, but also to make the use of natural resources for the biological control of pests to secure food production and security a possible alternative.

Cellulitis lesions in broiler chickens are induced by Escherichia coli Vacuolating Factor (ECVF).

Escherichia coli Vacuolating Factor (ECVF) is a heat-labile, vacuolating cytotoxin produced by avian pathogenic E. coli (APEC) isolated from avian cellulitis lesions. In this report, we intend to demonstrate that purified ECVF induces the inflammatory process of cellulitis. Our group is the first to demonstrate the effect of ECVF in a histological analysis by in situ inoculation of broiler chickens with purified ECVF. The animals were inoculated with the APEC AC53 and with purified ECVF subcutaneously on their ventral surface (in the sternum region). The histological analysis showed different grades of an acute inflammatory response in the epidermis, dermis and panniculus. An increase in mRNA expression of the proinflammatory cytokine TNF-α was also demonstrated in the inflamed tissue. When ECVF was systemically administered, increased levels of TNF-α and IL-10 were observed in the serum. These results suggest that ECVF plays a key role in the inflammatory process associated with cellulitis that is mainly mediated by TNF-α. In addition, this inflammation can be downregulated by the anti-inflammatory cytokine IL-10.

Exigência de metionina + cistina para frangas de reposição na fase recria de 13 a 18 semanas de idade / Requirement of methionine plus cystine for egg pullets in the growing phase from 13 to 18 weeks old

Determinou-se a exigência de metionina + cistina para frangas de 13 a 18 semanas de idade e verificou-se seu efeito sobre a fase de postura. O experimento foi dividido em duas fases experimentais, recria e postura. Na fase de recria, 560 aves - 50% Lohmann LSL e 50% Lohmann Brown -, com 13 a 18 semanas de idade, foram distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 5x2 (porcentagem de met+cis e linhagens), com quatro repetições e 14 aves por repetição. As porcentagens de met+cis totais estudadas foram 0,399; 0,469; 0,539; 0,609 e 0,679%. Ao atingirem 22 semanas de idade, 240 aves foram utilizadas para se avaliar o efeito residual das dietas oferecidas na fase de recria sobre o desempenho das aves, a produção e a qualidade de ovos. Ao se considerar a fase de produção, não se verificou efeito da inclusão de níveis de met+cis na ração, para ambas as linhagens. As exigências de metionina + cistina totais foram de 0,679% (0,611% de met+cis digestível) para aves leves e de 0,646% (0,581% de met+cis digestível) para aves semipesadas.

The methionine + cystine requirement was determined for pullets from 13 to 18 weeks of age and its effect was verified on the laying phase. The experiment was divided into two experimental phases. In the growing phase 560 birds (50% and 50% Lohmann LSL Lohmann Brown) at 13 to 18 weeks of age were distributed in a completely randomized factorial scheme 5x2 (methionine + cystine levels and lineages), with four replications and 14 birds per repetition. The levels of total methionine + cystine studied were 0.399, 0.469, 0.539, 0.609 and 0.679%. When they reached 22 weeks of age, 240 birds were used to assess the residual effects of the diets offered in the growing phase on performance of poultry, egg production and quality. Considering the production phase, there was no effect of the inclusion of methionine + cystine levels in the diet for both strains. The total methionine and cysteine was 0.679% (0.611% of digestible methionine + cystine) for light birds and 0.646% (0.581% methionine + cystine digestible) for brown-egg pullets.

Exigência de metionina + cistina para frangas de reposição na fase cria de sete a 12 semanas de idade / Requirement of methionine + cystine for pullets in the growing phase from 7 to 12 weeks old

Determinou-se a exigência de metionina + cistina para frangas de sete a 12 semanas de idade e verificou-se seu efeito sobre a fase de postura. O experimento foi dividido em duas fases, de cria e de produção, esta de 22 a 33 semanas de idade. Na primeira fase, utilizaram-se 640 aves - 50% Lohmann LSL e 50% Lohmann Brown - com sete a 12 semanas de idade, distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 5x2 (porcentagem de met+cis e linhagem), com quatro repetições e 16 aves por repetição. As porcentagens de met+cis total estudadas foram 0,471; 0,541; 0,611; 0,681 e 0,751%. Ao atingirem 22 semanas de idade, 240 aves foram utilizadas para verificar o efeito residual das dietas fornecidas na fase de cria. A dieta fornecida nesta fase foi igual para todas as aves. As exigências de metionina + cistina para aves de reposição, leves e semipesadas, no período de sete a 12 semanas de idade, foram de 0,710 % de met+cis total (0,639% de met+cis digestível) e 0,706% de met+cis total (0,635% de met+cis digestível), respectivamente.

The methionine + cystine requirement was determined for pullets from seven to 12 weeks of age and its effect was verified on the laying phase. The experiment was divided into two phases, growing and production, which lasted from 22 to 33 weeks of age. In the first experimental phase we used 640 birds (50% and 50% Lohmann LSL Lohmann Brown) allotted in a completely randomized factorial scheme 5x2 (methionine + cystine levels and lineages), with four replications and 16 birds per repetition. The levels of total methionine + cystine studied were 0.471; 0.541; 0.611; 0.681 e 0.751%. When they reached 22 weeks of age, 240 birds were used to verify the residual effect of the diets fed during the growth period. The level of methionine + cystine for white-egg and brown-egg pullets in the period from 1 to 6 weeks of age were 0.710% of total methionine + cystine (0.639 % met+cis digestible) and 0.706 % of total methionine + cystine (0.635 % of met+cis digestible), respectively.

Neuroproteomics: an insight into ALS.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of unknown aetiology. Diagnosis is made through physical examination, electrophysiological findings, and by excluding other conditions. There is not a single biomarker that concludes the diagnosis. The aim of this study was to investigate differentially expressed proteins in cerebrospinal fluid (CSF) of ALS patients compared to control subjects, with the purpose to identify a panel of possible biomarkers for the disease. The differentially expressed spots/proteins were submitted to two-dimensional (2D) electrophoresis and recognized with matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Parkin-like and many iron and zinc binding were some of the proteins found in ALS CSF. Parkin is a ligase involved in ubiquitin-proteasome pathway and mutations in the parkin gene are the most common cause of recessive familial Parkinson's disease. Iron and zinc are involved with many important metabolic processes and are related to neurodegenerative disease. Common features of ALS comprise failure of the ubiquitin-proteasome system and increased levels of metal ions in the brain. Therefore, the identification of these proteins can be a significant step in ALS research. These and other identified proteins are discussed in this study.

Comparison of ultrasonography and histopathology for the diagnosis of endometritis in Holstein-Friesian cows.

Reproductive diseases are a great barrier to dairy production, as they diminish fertility and cause economic loss. In this context, endometritis plays an important role. Different diagnostic techniques exist for endometritis that vary in sensitivity and specificity. This work aimed to evaluate and compare ultrasonographic and histopathological examinations, and combinations thereof, for the diagnosis of endometritis by determining their sensitivity, specificity, and kappa agreement coefficients using endometrial cytology as the gold standard. We developed an adaptive reading score to perform histological examination of the uterus. In this study, 76 Holstein-Friesian cows, 21 to 47 d postpartum, without vaginal catarrh were submitted to gynecological examinations, including ultrasonographic examination, an endometrial cytology examination, and a uterine biopsy. Our results indicate that ultrasonography is a practical and effective diagnostic method, and a combination of the determination of intrauterine fluid and cervical diameter was the most efficient ultrasonographic method, with 50% sensitivity and 88% specificity. Histopathological examination was a useful diagnostic method that provided rich detail on endometrial inflammation, and allowed evaluation of different components of the tissue such as the epithelium, lamina propria, endometrial glands, and blood vessels; this method had 44% sensitivity and 92% specificity for diagnosis of endometritis.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.

Fiber fermentability effects on energy and macronutrient digestibility, fecal traits, postprandial metabolite responses, and colon histology of overweight cats.

Considering the different potential benefits of divergent fiber ingredients, the effect of 3 fiber sources on energy and macronutrient digestibility, fermentation product formation, postprandial metabolite responses, and colon histology of overweight cats (Felis catus) fed kibble diets was compared. Twenty-four healthy adult cats were assigned in a complete randomized block design to 2 groups of 12 animals, and 3 animals from each group were fed 1 of 4 of the following kibble diets: control (CO; 11.5% dietary fiber), beet pulp (BP; 26% dietary fiber), wheat bran (WB; 24% dietary fiber), and sugarcane fiber (SF; 28% dietary fiber). Digestibility was measured by the total collection of feces. After 16 d of diet adaptation and an overnight period without food, blood glucose, cholesterol, and triglyceride postprandial responses were evaluated for 16 h after continued exposure to food. On d 20, colon biopsies of the cats were collected under general anesthesia. Fiber addition reduced food energy and nutrient digestibility. Of all the fiber sources, SF had the least dietary fiber digestibility (P < 0.05), causing the largest reduction of dietary energy digestibility (P < 0.05). The greater fermentability of BP resulted in reduced fecal DM and pH, greater fecal production [g/(cat × d); as-is], and greater fecal concentration of acetate, propionate, and lactate (P < 0.05). For most fecal variables, WB was intermediate between BP and SF, and SF was similar to the control diet except for an increased fecal DM and firmer feces production for the SF diet (P < 0.05). Postprandial evaluations indicated reduced mean glucose concentration and area under the glucose curve in cats fed the SF diet (P < 0.05). Colon mucosa thickness, crypt area, lamina propria area, goblet cell area, crypt mean size, and crypt in bifurcation did not vary among the diets. According to the fiber solubility and fermentation rates, fiber sources can induce different physiological responses in cats, reduce energy digestibility, and favor glucose metabolism (SF), or improve gut health (BP).