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Apesar do diagnóstico da anemia infecciosa e mais evidências recentes de infecção em campo (Brentano & Ottati, 1999), não havia até hoje dados disponíveis da prevalência do vírus da anemia das galinhas (CAV) no Brasil. Uma vez que o conhecimento da prevalência do vírus e da distribuição de anticorpos nos lotes de matrizes é essencial para o delineamento de estratégias de controle da doença, realizamos um levantamento em 5 Estados com produção comercial intensiva de linhas de corte, abrangendo lotes de aves com diferentes idades. De 2.355 soros testados por ELISA, 89% foram positivos, indicando a alta prevalência do CAV no Brasil, em todos os Estados testados. A análise do soro de aves das 6 até as 70 semanas de idade demonstrou que lotes de 6 a 18 semanas apresentam desuniformidade na imunidade ao CAV, caracterizada por títulos de anticorpos abaixo dos níveis considerados protetores e aves ainda negativas para o CAV. A desuniformidade nos títulos de anticorpos e a ausência dos mesmos são evidentes ainda, mas em menor proporção também até as 24 semanas, enquanto que apenas partir das 25 semanas há 100% de soroconversão, acompanhada de significativa redução da desuniformidade no título de anticorpos das matrizes para menos de 2,5% das aves. Esses resultados fornecem evidência de uma progressiva disseminação do vírus em matrizes de corte, mas com riscos da presença de aves ainda susceptíveis à transmissão vertical e inadequada transferência de imunidade passiva no início do período de postura, que podem culminar em surtos da doença na progênie.

Salmonella Enteritidis remains a leading cause of human foodborne disease, mostly associated with the consumption of contaminated poultry products. To more strategically implement a phage therapy scheme for S. Enteritidis control in broilers, a cocktail containing three wild-type lytic bacteriophages (LBs) previously isolated from chickens was evaluated shortly and later after a challenge. Genomic characterization, lytic spectrum and in vitro efficacy were determined for each studied LB. In independent trials, broilers challenged with S. Enteritidis on day of hatch received phage therapy from 6 to 10 days of age (early treatment), and from 31 to 35 days of age (later treatment). S. Enteritidis analyses were performed before treatment and at 1, 4, 7 and 10 days post-treatment (dpt) in both trials. Partial DNA sequence analysis of each LB revealed close similarity to the Ackermannviridae family. LBs lysed different Salmonella enterica serovars, while other tested bacteria were refractory. An in-vitro reduction of 1.49, 0.65 and 0.58 log 10 CFU/mL in S. Enteritidis number was obtained after co-incubation for 3 h with each LB. Both in vivo trials showed a significant reduction in the average number of intestinal S. Enteritidis calculated after phage therapy compared with controls. However, the highest efficiency was found in the later therapy, which resulted in a reduction of 1.08 log 10 CFU/g in the average from 4 to 10 dpt, showing potential for future use as a pre-harvest strategy to reduce the S. Enteritidis intestinal colonization in broilers on farms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Housekeeping genes (HKG) are genes necessary for the maintenance of basal cellular functions, regardless of the specific roles within a tissue. It, therefore, is expected that these genes will maintain a relatively constant expression profile when there are varying physiological conditions. The identification of tissue specific reference genes is highly important for the normalization of gene expression profiles among different tissues. In this sow study, the objective was to identify stable reference genes in the uterine tissue and corpus luteum (CL), 6 days post-artificial insemination. The stability of ubiquitin (UBB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), DNA topoisomerase 2-beta (TOP2B), histone H3 (H3F3A) and hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1) abundances of mRNA were evaluated using the Bestkeeper technique. Briefly, total RNA was extracted for each tissue from 20 gilts (n = 20), processed by RT-qPCR and submitted to analysis using the Bestkeeper technique, which allowed for the evaluation of the consistency in abundance of mRNA for the reference genes. For all evaluated genes, the abundance of mRNA was relatively consistent in the uterine tissue, with the greatest abundance being for the GAPDH and TOP2B genes. The analysis of these genes in the CL, however, indicated there was a relatively greater variation of mRNA abundance for the various reference genes. Data suggest that UBB was the reference gene with the most consistent relative abundance of mRNA and that this gene could be used as a reference for corpora lutea analyses of mRNA., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The microbiological risk of recycled litter depends on the efficacy of the management system applied to inactivate residual microorganisms and preserve the health of the successive broiler flock. This study aimed to assess the viability and infectivity of the Newcastle Disease Virus (NDV), Infectious Bursal Disease Virus (IBDV) and Salmonella Heidelberg in recycled litter exposed to different treatments. The litter was contaminated with microorganisms and submitted to the treatments (T): T1: shallow fermentation; T2: quicklime (calcium oxide); T3: shallow fermentation followed by addition of quicklime; T4: no treatment. Sentinel chicks housed on the treated litter showed that T1 and T3 inactivated residual IBDV. Analysis of the litter subjected to T1 also showed reduced levels of total enterobacteria. T2 was not able to reduce the microorganisms assessed and its association with T1 (T3) failed to enhance the effect of the treatment. NDV did not survive in the broiler litter, regardless of the treatment applied, and it was also not detected in the sentinel chicks. S. Heidelberg remained viable in the litter submitted to all studied treatments, being isolated from the sentinel chicks of all the experimental groups. The antimicrobial activity of T1 and T3 was associated to higher ammonia contents in the broiler litter. The results indicate that the shallow fermentation treatment is efficient for controlling residual IBDV and total enterobacteria in the recycled litter., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Avian infectious bronchitis virus (IBV) primarily replicates in epithelial cells of the upper respiratory tract of chickens, inducing both morphological and immune modulatory changes. However, the association between the local immune responses induced by IBV and the mechanisms of pathogenesis has not yet been completely elucidated. This study compared the expression profile of genes related to immune responses in tracheal samples after challenge with two Brazilian field isolates (A and B) of IBV from the same genotype, associating these responses with viral replication and with pathological changes in trachea and kidney. We detected a suppressive effect on the early activation of TLR7 pathway, followed by lower expression levels of inflammatory related genes induced by challenge with the IBV B isolate when compared to the challenge with to the IBV A isolate. Cell-mediated immune (CMI) related genes presented also lower levels of expression in tracheal samples from birds challenged with B isolate at 1dpi. Increased viral load and a higher percentage of birds with relevant lesions were observed in both tracheal and renal samples from chickens exposed to challenge with IBV B isolate. This differential pattern of early immune responses developed after challenge with IBV B isolate, related to the downregulation of TLR7, leading to insufficient pro-inflammatory response and lower CMI responses, seem to have an association with a most severe renal lesion and an enhanced capability of replication of this isolate in chicken.

Newcastle disease virus (NDV) causes a fast-spreading, highly contagious infectious disease in several bird species. Commercial poultry farms in Brazil were considered free of virulent NDV. Data on NDV infection levels in backyard poultry flocks and the epidemiology of the disease are limited. The aim of this study was to perform a NDV survey in backyard poultry from households flocks located around one of the main wintering sites for migratory wild birds in Brazil, and to identify potential risk factors associated with NDV. Backyard poultry may be sentinels and a source of infection for commercial poultry, since they may have as much contact with these birds as with migratory wild birds. Data were collected from 48 randomly selected households using an epidemiological questionnaire. Serum samples from poultry were tested for NDV antibodies using an ELISA, and tracheal and cloacal swabs were collected for NDV molecular detection. The risk factors were assessed using a multivariate Poisson regression with robust variance. The ELISA showed that 33.8% of the serum samples were positive for anti-NDV antibodies and in 42 households (87.5%) at least one NDV-positive bird was found. Tracheal and cloacal swabs were negative for NDV by real time RT-PCR, possible because within this region there might flow a low pathogenicity NDV strain, which can induce seroconversion with innaparent clinical findings. The prevalence ratio (PR) increased when farmers used their own replacement poultry to restock their flock (PR=1.64; 95% CI: 1.11-2.42). Furthermore, the increasing distance of the household flock from the "Laguna do Peixe" estuary was associated with decreasing NDV seropositivity (PR=0.94; 95% CI: 0.90-0.99). This is the first study in Brazil evaluating the presence of NDV and the associated risk factors in households with backyard poultry flocks. The great number of farms with seropositive birds indicates that the virus circulates in backyard flocks, and this breeding system may be a source of NDV. These data can be used to establish appropriate biosecurity and husbandry measures for this type of breeding system to prevent NDV spread in Brazil., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Here we report the isolation of Newcastle disease virus (NDV) from cloacal swabs obtained from penguins in the South Atlantic Antarctic region (62°08S, 58°25W). Samples of 100 penguins from King George Island were tested by real-time PCR, of which 2 (2%) were positive for NDV. The positive samples were isolated in embryonated chicken eggs and their matrix and fusion proteins genes were partially sequenced. This was complemented by the serological study performed on the blood of the same specimens, which resulted in a 33.3% rate of positivity., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Purification of chicken anemia virus (CAV) VP3 protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. CAV particle was obtained from infected liver of chicken and DNA was extracted. The VP3 protein gene was amplified from the extracted DNA by polymerase chain reaction (PCR) and cloned. The recombinant expression construct (pTrc-VP3) was identified by PCR and sequencing analysis. Expression of VP3 protein with a molecular mass of approximately 21kDa was confirmed by Western blotting analysis with CAV-specific antibodies. The in vitro expressed VP3 protein was purified to near homogeneity by elution from the gel, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The purified VP3 protein was recognized by CAV antibodies in a Western blotting assay. This finding indicates that recombinant VP3 expressed in the pTrcHis2 vector system can be used as antigen to detect anti-CAV antibodies.

Specific amino acid (aa) substitutions in VP1, VP2 and VP3 genes were reported as a distinctive feature of the American CIA-1 strain, characterized as having a variable rate of growth and tropism for different MSB-1 cell sublines [Renshaw RW, Soiné C, Weinkle T, O'Connell PH, Ohashi K, Watson S, et al. A hypervariable region in VP1 of chicken anemia virus mediates rate of spread and cell tropism in tissue culture. J Virol 1996;70(12):8872-8]. DNA sequencing of 878 nucleotides from twelve Brazilian CAV, eight of which tested for in vitro isolation in three different sources of MDCC-MSB1 cell line and identified as lacking capacity to propagate in any of these cells, were compared to sequence data available for CAV strains propagated or not in cell culture. Alignment of the deduced aa resulted in a lack of singled out amino acid substitutions in the partial genomic sequences of Brazilian isolates that would entirely contrast them to viruses propagated in MSB-1 cells, indicating that the combined VP1, VP2 and VP3 substitutions observed may not entirely account as sole determinants of CAV isolation and propagation in MDCC-MSB-1 cells.

Influenza A and Newcastle disease viruses are pathogens of social and economical importance known to be disseminated throughout the world by migratory birds. Many efforts have been made to control the introduction of these viruses into new environments, and complete world surveillance has yet to be achieved. Virus isolation and immunofluorescence techniques are time consuming, have inherent limited sensitivity and present a lack of host cells permissive universally to all Influenza A viruses. In this paper, the use of a duplex RT-PCR is described capable of sorting out any NDV and Influenza A virus strain simultaneously in oral and cloacal swab specimens. This method includes fluorescent detection of amplicons that provide accurate analysis of many DNA fragments within one base discrimination. Reference viruses were used for standardization of the assay and samples from wild-type viruses were screened, with four positive results for Influenza A detected in migratory birds captured in the state of Sao Paulo. This screening test can be considered a first step for further studies of these viruses circulating in avian species in Brazil, and hopefully will contribute to broaden the sample spectrum from wild birds, leading to a better understanding of these viruses and their participation in the southeastern region of the country.

Background/aims: Protein-calorie malnutrition that occurs in patients with squamous cell carcinoma of the esophagus presents defined implications in surgical results. The present study aims to establish a nutritional score for malnutrition to measure the prevalence of protein-calorie malnutrition and to determine parameters with higher discriminatory power for protein-calorie malnutrition diagnosis., Methodology: Forty-five (45) patients with squamous cell carcinoma of the esophagus (cases) and 90 surgical patients (controls) were analyzed using twelve parameters: percentage of body weight loss, triceps and subscapular skin fold, mid arm circumference and arm muscle circumference, serum total proteins, albumin, transferin, pre-albumin, hematocrit, total lymphocyte count and late hypersensitivity skin tests. Prognostic Nutritional Index was calculated for all patients., Results: There was a statistically significant difference between the two groups (p < 0.05) regarding all evaluated parameters except late hypersensitivity skin tests. Patients were classified as malnourished when presented with six or more altered parameters. Through the proposed score, the prevalence of protein-calorie malnutrition was defined in cases (57.8%) and in controls (7.8%) groups with statistic significant difference (p = 0.001)., Conclusions: The proposed score allowed the diagnosis of malnourished patients, defined the malnutrition prevalence in both groups and established parameters with a greater discriminatory power for the protein-calorie malnutrition diagnosis.

To determine long-term survival of patients with thoracic oesophageal tumor who underwent resection, and to identify possible associated prognostic factors, 58 patients underwent oesophagectomy alone (group A) and 16 combined with neo-adjuvant chemoradiotherapy (group B). Univariate and multivariate analysis of prognostic factors were performed for age, depth of oesophageal wall tumour penetration, node involvement, type of resection, TNM stage and degree of tumour differentiation. Long-term survival rates at 1-5 years were 81% versus 89%; 56% versus 67%; 30% versus 67%; 12% versus 44%; and 0% versus 33% for group A and B, respectively (P = 0.0543, NS). Univariate analysis revealed only depth of invasion (P = 0.0076) and TNM stage (P = 0.0452) as isolated prognostic factors for long-term survival and multivariate analysis did not demonstrate any independent factor. Despite the small number of cases, neoadjuvant chemoradiotherapy seems to improve prognosis as well as to allow resection in a greater number of cases due to tumor downstaging.

A retrospective analysis on the clinical-surgical handling of patients with enterocutaneous fistula (ECF) was performed, where an alternative surgical technique was discussed: intestinal bypass. Fistula with draining over 500 ml/24 h, which were present in 13 patients, were classified as high debit. We defined as complex, the fistula with multiple orifices, high defect of the abdominal wall or through the mesh. The population studied consisted of 25 patients, 11 male, in a total of 34 ECF and mean age of 41.9 years. At clinical treatment with TPN for high debit ECF, 2 patients (16.6%) were cured, another 2 died and 8 (66.8%) needed surgical treatment. The surgery cured 7 patients (77.7%) with high debit ECF but 2 (22.3%) died. In the patients with low debit ECF, TPN cured 2 patients (40%) but failed in another 3 (60%). All patients with low debit ECF resolved with surgical treatment.

Mesenteric cysts are rare intra-abdominal tumors with an incidence around one case per 100,000 hospital admissions. The clinical presentation is variable; patients may be asymptomatic or present with either acute or chronic abdominal pain. Physical examination commonly demonstrates a smooth, round and mobile abdominal mass. Differential diagnosis includes any abdominal cyst or tumor. Laboratory tests are usually helpless. Ultrasonography and CT scans are the best diagnostic tools. The treatment of choice is the total resection of the cyst, which is regularly performed by open surgery. This paper reports a case of a mesenteric cyst successfully resected by laparoscopy, and addresses the possible uses of this approach.

Introduction: The esophageal epidermoid cancer has an important association with aerodigestive tract neoplasms and possibly share the same risk factors. Furthermore, other neoplasms can be associated with esophagus cancer., Objective: To analyze retrospectively the patients with esophageal epidermoid cancer (EEC) and associated neoplasms, treated by the Esophagus Stomach and Small Intestine Group of Surgery at Hospital de Clinicas de Porto Alegre from January 1988 to June 1995., Patients and Methods: Nineteen (7.28%) of the 261 studied patients had associated neoplasms to the EEC. Ten patients presented synchronic tumours and 9 metachronic ones. The predominant sex was the masculine with 17 cases. The mean age was 62.52 years in the moment of the esophageal cancer diagnostic., Results: The aerodigestive tumours, squamous carcinomas in totality, represented the predominant associated neoplasm histological type in 68.42% of the cases. The most frequent associated aerodigestive tumours site was the respiratory tract (53.8%), followed by the oral cavity and oropharynx (23%) and larynx (23%). In our sample, twelve patients were smokers and 9 were alcohol abusers. In relation to the EEC treatment, surgery was performed in 6 patients. The associated neoplasm was treated with radical surgery in 11 patients and radiotherapy in 5. Surprisingly 4 cases (21%) of gastric adenocarcinoma associated to the EEC were diagnosed, treated with radical surgery in 3 patients., Conclusion: The authors call attention to the importance of a criterial staging as well as the follow up in patients with EEC owing to the significant association with others neoplasms, principally with aerodigestive tumours, and discuss the common risk factors possibility: tobacco and alcohol use. Important association with gastric neoplasms were found in this casuistry.

Background: The surgical approach for patients with advanced epidermoid esophageal carcinoma should provide an effective palliative effect with morbidity ratio as low as possible. Anastomotic leakage is a frequent complication and may be responsible for both early and late morbidity and, therefore, we assessed the role of delayed cervical esophagovisceral anastomosis technique in relation to the incidence of anastomotic complications., Methods: Eight patients (Group 1) and 12 patients (Group 2) submitted to one-stage or two-stage operation, respectively, were selected by an intraoperative assessment by the surgeon, considering mainly tissue blood flow of the replacement organ after its placement in the cervical region., Results: In Group 1 anastomotic dehiscence was observed in 37.5% of patients, while in Group 2 no cases of dehiscence occurred (p = 0.049). However, the postoperative mortality rate did not differ between the two groups (12.5% versus 0%, NS)., Conclusion: When organ viability is uncertain, esophagovisceral anastomosis is best done by two-stage operation, since it decreases the incidence of anastomotic leak.

Seven chicken infectious anemia virus (CIAV) isolates were obtained from seven broiler flocks with poor performance in two states of Brazil. All isolates induced thymus atrophy, bone-marrow aplasia, and low hematocrit values when inoculated into 1-day-old susceptible chicks. The CIAV isolates were resistant to treatment with chloroform and were able to pass through 50-nm-pore-size filters. CIAV-specific antigens could be demonstrated in tissues of experimentally infected chicks using a monoclonal antibody specific for CIAV. These characteristics of the virus and the virus-induced lesions demonstrate that CIAV is present in Brazil and that the virus is associated with production problems.

1. The Bartha K and NIA-4 strains of Aujeszky's disease virus (ADV) were non-pathogenic for rabbits vaccinated once or twice by nasal instillation or intramuscular injection. Neutralizing antibodies were detected in 68% of the rabbits two weeks after primary vaccination and in all rabbits at challenge. 2. Challenge doses of virulent ADV greater than 10(5.0) median tissue culture infective doses (TCID50) resulted in the death of most vaccinated and all unvaccinated rabbits with typical signs of Aujeszky's disease within 4 days. ADV was recovered from brain and lung suspensions of vaccinated and unvaccinated rabbits who had died as a result of the challenge. 3. When the challenge dose was reduced to approximately 10(3.0) TCID50, rabbits vaccinated twice survived while all unvaccinated controls died within 3 days.