Repositório RCAAP

CONTEXT: Several studies have demonstrated a higher prevalence of celiac disease (CD) among females with Turner syndrome when compared to the general population. Nevertheless, there is no record in literature concerning this investigation among Brazilian patients. OBJECTIVE: To assess the prevalence of CD among a group of Brazilian patients with Turner syndrome. METHODS: Fifty-six females with Turner syndrome and on gluten-containing diet were screened for CD utilizing immunoglobulin A antiendomysium (IgA-EMA) and immunoglobulin A anti-tissue transglutaminase (IgA-tTG) antibody assays. Additionally, they were genotyped for CD human leukocyte antigen (CD-HLA) predisposing alleles. Patients showing positivity in serological testing were offered to perform small intestine biopsy for histological confirmation. RESULTS: Mean age at diagnosis of Turner syndrome was 5.5 ± 4.4 years; mean age at screening for CD was 17.0 ± 9.3 years (from 10 months of age to 52 years). Two girls were positive for IgA-EMA and IgA-tTG, presented predisposing HLA-DQ2 alleles and both had the diagnosis of CD confirmed by jejunal biopsy. CONCLUSION: The 3.6% prevalence of biopsy-proven CD among this group of females with Turner syndrome is 10 times higher than the one among females from the general population of the same geographical area. This result provides additional support to an association between these two disorders and restates that girls and women with Turner syndrome represent a high risk population for developing CD.

CONTEXT: Patients with autoimmune rheumatologic conditions and celiac disease tend to have a variety of autoantibodies, many of which have no clear pathogenic role. The literature contains frequent reports of celiac disease being more prevalent in patients with rheumatologic diseases, although this remains controversial. OBJECTIVES: To investigate the prevalence of positive serum tests for celiac disease, particularly IgA and IgG antigliadin (AGA) antibodies and IgA antiendomysium antibodies (EmA) in patients with autoimmune rheumatologic diseases. A second aim was to correlate positive serum tests with prednisone and immunosuppressant medication. METHODS: A total of 190 adults and pediatric patients with a variety of autoimmune rheumatologic diseases (systemic lupus erythematosus, rheumatoid arthritis, juvenile rheumatoid arthritis and spondyloarthrophathies) were evaluated and tested for IgA and IgG antigliadin-antibodies and IgA antiendomysium antibodies. Patients with positive serum tests underwent endoscopic duodenal biopsies for pathology studies. RESULTS: There were four positive sera (2.1%) for AGA IgA, all of which tested negative for AGA IgG and EmA. Three sera (1.6%) tested positive for AGA IgG; all were negative for AGA IgA and EmA. The EmA test at a 1:2.5 serum dilution tested positive in 94 patients (49.5%); at a 1:5 serum dilution it was positive in 41 patients (21.6%). Eleven subjects tested positive for EmA at 1:40 dilution; and all of these tested negative for IgA tissue antitransglutaminase (tTG) antibodies. Nine of the 11 EmA-positive patients and all 7 patients with positive antigliadin antibodies tests underwent duodenal endoscopic biopsies, and no significant changes were demonstrated in their duodenal mucosa. A positive EmA was associated with elevated optical density AGA IgA readings; however, there was no relationship between positive EmA and AGA IgG optical density readings. Prednisone and immunosuppressant use were unrelated to AGA IgA optical density readings or AGA IgG readings. These drugs were associated with fewer positive EmA tests. CONCLUSIONS: Positive AGAA, AGAG or EmA results are probably nonspecific for the presence of celiac disease among autoimmune rheumatologic disease patients. The intake of prednisone and immunosuprressant drugs seems to reduce the prevalence of IgA EmA, but it does not interfere with antigliadin antibodies tests.Further studies are required to estimate more accurately the prevalence of this disease in rheumatologic patients.

CONTEXT: Celiac disease susceptibility has been shown to be associated with the HLA alleles DQA1*0501 and DQB1*0201 (together encoding the DQ2 heterodimer) that are present in practically all celiac disease patients. The DQ8 heterodimer (coded by DQA1*03-DQB1*0302), which is carried on a DRB1*04 (DR4) haplotype, is commonly encoded for by the few celiacs who do not carry the DQ2 heterodimer. Only a few celiac disease patients have been reported without these known risk alleles. OBJECTIVE: To assess the prevalence of celiac disease in a group of first degree relatives of celiac patients, and the frequency of HLA predisposing alleles both in the group of celiac patients and in their first degree relatives, identifying those first degree relatives who would need further screening for celiac disease. METHODS: Ninety celiac disease patients and 207 first degree relatives underwent serologic screening for celiac disease (endomysial and transglutaminase antibodies) followed by intestinal biopsy in positive patients. The HLA-DQA1*0501, DQB1*0201 and DRB1*04 frequencies of celiac patients and their first degree relatives were determined utilizing the PCR method. RESULTS: All the celiac disease patients (n = 90) with the exception of one (1.1%) carried at least one of the alleles investigated. Altogether 11 (5.3%) of the investigated first degree relatives did not carry any of the alleles studied. Fourteen (6.7%) new cases of celiac disease were found among the 207 celiac disease patients first degree relatives. CONCLUSIONS: Considering the cost-benefit of the HLA typing of all the first degree relatives of celiac patients, their HLA status should be decided on an individual basis, taking account of their profile and preferences, and the existence of other medical conditions.

CONTEXT: The surgical treatment for morbid obesity is becoming common in this country. Only a few papers reported the long-term results of the surgical approach for morbid obesity, mainly in terms of quality of life. OBJECTIVE: To compare mortality rate, weight loss, improvement of both diabetes and hypertension, and quality of life of patients from the public healthcare in Cuiabá, MT, Brazil, who underwent either medical or surgical interventions after a minimum of 2 years. METHODS: The population of this study was constituted by morbidly obese patients who initiated treatment between June 2002 and December 2006. The casuistic consisted of 89 patients submitted to medical therapy and 76 patients who underwent surgical procedures. The main variables were weight loss, improvement of hypertension and diabetes, quality of life, and mortality. RESULTS: The overall results showed that weight loss was significant in the two groups (P<0.001); however surgical patients showed a greater loss than the medical group (P = 0.05). The improvement of diabetes and hypertension was significantly greater in the surgical group (P<0.001), in which no cases of diabetes persisted. There was an increase in cases of hypertension among patients receiving medical attention. Mortality occurred in six cases (6.7%) of the medical group and in five cases (6.6%) of the surgical group (P = 0.97). The median grade of the quality of life score obtained by surgical patients (2.37 [range: -2.50 to 3.00]) was significantly greater (P<0.001) when compared to the medical group (1.25 [range: -1.50 to 3.00]). CONCLUSION: The surgical group presented better results regarding the weight loss, quality of life and improvement of hypertension and diabetes. There was no significant difference in mortality rate between the two groups after a minimum of 2 years.

CONTEXT: Recent studies have shown that local anesthesia for loop colostomy closure is as safe as spinal anesthesia for this procedure. OBJECTIVES: Randomized clinical trial to compare the results from these two techniques. METHODS: Fifty patients were randomized for loop colostomy closure using spinal anesthesia (n = 25) and using local anesthesia (n = 25). Preoperatively, the bowel was evaluated by means of colonoscopy, and bowel preparation was performed with 10% oral mannitol solution and physiological saline solution for lavage through the distal colostomy orifice. All patients were given prophylactic antibiotics (cefoxitin). Pain, analgesia, reestablishment of peristaltism or peristalsis, diet reintroduction, length of hospitalization and rehospitalization were analyzed postoperatively. RESULTS: Surgery duration and local complications were greater in the spinal anesthesia group. Conversion to general anesthesia occurred only with spinal anesthesia. There was no difference in intraoperative pain between the groups, but postoperative pain, reestablishment of peristaltism or peristalsis, diet reintroduction and length of hospitalization were lower with local anesthesia. CONCLUSIONS: Local anesthesia plus sedation offers a safer and more effective method than spinal anesthesia for loop colostomy closure.

CONTEXT: Upper gastrointestinal bleeding associated to esophageal varices is the most dramatic complication of cirrhosis. It is recommended screening every cirrhotic for esophageal varices with endoscopy. OBJECTIVES: To evaluate the capacity of the platelet count/spleen diameter ratio in non-invasively predicting esophageal varices in a population of cirrhotics originated in an independent center from the one in which it was developed. METHODS: The study included patients from the ambulatory care clinic of cirrhosis of a Brazilian hospital and studied platelet count, spleen diameter and presence of esophageal varices, as well as Child and MELD scores. It used a cutoff value of 909 for the platelet count/spleen diameter ratio, as previously published. A sample of 139 patients was needed to grant results a 95% confidence level. RESULTS: The study included 164 cirrhotics, 56.7% male, with a mean age of 56.6 ± 11.6 years. In the univariate analysis, platelet count, spleen diameter, presence of ascites, Child and MELD scores and the platelet count/spleen diameter ratio were related to esophageal varices (P<0.05). The platelet count/spleen diameter ratio had sensitivity of 77.5% (95% CI = 0.700-0.850), specificity of 45.5% (95% CI = 0.307-0.602), positive predictive value of 79.5% (95% CI = 0.722-0.868), negative predictive value of 42.6% (95% CI = 0.284-0.567) and accuracy of 68.9% (95% CI = 0.618-0.760). In the multivariate analysis, platelet count was the only variable which related to esophageal varices (P<0.05). CONCLUSION: Platelet count/ spleen diameter ratio is not adequate to predict esophageal varices in cirrhotics.

CONTEXT: Crohn's disease presents very heterogeneous features from a clinical point of view, and classifying Crohn's disease patients in homogeneous subgroups in the light of genetic, molecular and clinical aspects is challenging. The Montreal Classification for Crohn's disease was proposed in 2005 as an effort to characterize Crohn's disease patients according to recent clinical and research advances in the field of inflammatory bowel disease. Since its proposition, the Montreal Classification needs an ample validation and application among different populations around the world. To date, there are no known studies applying the Montreal Classification to a Brazilian cohort of Crohn's disease patients. OBJECTIVES: To apply the Montreal Classification to a Brazilian cohort of Crohn's disease patients at a referral center for inflammatory bowel disease in Northwestern São Paulo State, Brazil. METHODS: We selected 90 consecutive well-characterized Crohn's disease patients assisted at Inflammatory Bowel Disease Outpatient Clinic between January 1992 and January 2007, with a minimum follow-up of 2 years; data concerning demographic characteristics, clinical onset of disease, age at diagnosis, time of disease, location and behavior of disease, presence of extraintestinal manifestations, familial occurrence, perianal involvement, treatment with biological drugs, and history of surgical treatment were evaluated. RESULTS: Male patients were predominant (54%), with a mean age at diagnosis of 33 ± 14 years old, and a median followup of 5.5 years. Most of the patients were diagnosed between 17 and 40 years old (59%), and had disease located in terminal ileum 46%), with a nonstricturing, nonpenetrating behavior (71%). Time of disease was correlated with necessity of biological treatment, disease behavior, and surgical treatment (P<0.05). CONCLUSIONS: These study findings are consistent with results from other studies conducted among different populations, although a further multicentric study with a larger number of patients would be necessary to validate the Montreal Classification among Brazilian population.

CONTEXT: Chronic diseases have an impact on the quality of life of the individuals. OBJECTIVE: To evaluate the quality of life of adults with inflammatory bowel disease by the inflammatory bowel disease questionnaire certified for Portuguese language. METHODS: We interviewed 50 individuals from both genders aged from 18 to 60 years old, with Crohn's disease and ulcerative colitis, regardless of disease activity. RESULTS: The average age of the sample was 42.2 ± 13.6 years old, the disease length was 98.8 ± 74.3 months, and 72% of patients had Crohn's disease and 14% presented disease activity. There was no significant difference in questionnaire scores of patients with different inflammatory bowel disease when they are in the remission phase (172.0 ± 42.4 and 173.6 ± 28.2 for ulcerative colitis and Crohn's disease, respectively, P = 0.886). When compared to scores of patients who were in crisis, it was found that they have a lower quality of life that patients in remission (123.8 ± 44.5 and 173.3 ± 31.5 for patients in crisis and remission, respectively, P = 0.001). CONCLUSION: It was noticed that the quality of life of patients with inflammatory bowel disease did not differ among patients with Crohn's disease or ulcerative colitis, when patients are in remission. The main aspect which determines the loss of quality of life would be being at the stage of disease activity.

CONTEXT: Gamma-aminobutyric acid (GABA) is a potent inhibitory neurotransmitter. There is evidence that GABA B receptors located in the dorsal complex and in afferent fibers of the vagus nerve participate in the control of gastrointestinal motility. OBJECTIVE: To assess the intracerebroventricularly (ICV) and intravenously (IV) effect of baclofen, a GABA B receptor agonist, on liquid and solid gastric emptying in rats. METHODS: Adult male Wistar rats weighing 250-300 g (n = 6-8 animals) were used. Gastric emptying of liquid test meals labeled with phenol red was evaluated by the determination of percent gastric retention (%GR) 10 and 15 min after orogastric administration of saline and 10% glucose meals, respectively. Baclofen was injected ICV (1 and 2 µg/animal) through a tube implanted into the lateral ventricle of the brain and was injected IV (1 and 2 mg/kg) into a tail vein. The gastric emptying of liquid was determined 10 or 30 min after ICV and IV baclofen administration, respectively. The gastric emptying of the solid meal was assessed by the determination of percent gastric retention 2 h after the beginning of the ingestion of the habitual ratio by the animal, consumed over a period of 30 min. Baclofen was administered ICV (1 and 2 µg/animal) or IV (1 and 2 mg/kg) immediately after the end of the ingestion of the solid meal. The control groups received vehicle (sterile saline solution) ICV or IV. RESULTS: The group of animals receiving baclofen ICV (2 mg/animal) presented a significantly lower (P<0.05, Tukey test) %GR (mean ± SEM) of the saline (18.1 ± 2.5%) compared to control (33.2 ± 2.2%). In the group receiving the drug IV, the gastric retention of the same test meal did not differ from control. ICV and IV administration of baclofen had no effect on the gastric emptying of the 10% glucose solution compared to control. ICV administration of 1 or 2 mg baclofen/animal significantly increased the gastric retention of the solid test meal (57.9 ± 6.5% and 66.6 ± 6.3%, respectively) compared to control (35.1 ± 4.4%). The same phenomenon was observed only with the IV dose of 2 mg/kg (71.9 ± 2.6%) compared to control (52.7 ± 2.8%). CONCLUSION: Baclofen administered: 1. ICV (2 µg/animal), but not IV, increased gastric emptying of a non-caloric isotonic liquid test meal (saline); 2. when administered ICV or IV, it had no effect of gastric emptying of a 10% glucose solution; 3) when administered ICV (1 and 2 mg/animal) and IV (2 mg/kg) it delayed the gastric emptying of the solid meal.

CONTEXT: There are consistent clues of worse results with grafts from older donors, especially in hepatitis C. University of Wisconsin (UW) solution is adopted for liver preservation, but other solutions are being studied, as fructose-1,6-bisphosphate (FBP). OBJECTIVE: To determinate the impact of aging of the donor on the cold ischemia injury in rat livers and compare UW and FBP. METHODS: Twenty male Wistar rats were studied - 10, aging 20 weeks: 5 to preservation with UW (C-UW) and 5, FBP (C-FBP); and other 10, aging 50 weeks: 5 to UW (E-UW) and 5 to FBP (E-FBP). Rats were anesthesized, submitted to hepatectomy, and graft was kept under 2-4ºC for 8 hours. Liquid samples were taken at 2, 4, 6, and 8 hours, to determine AST and LDH. At the end, in the liver tissue thiobarbituric acid reactive substances and catalase were determined. RESULTS: Average liver weight was similar between controls and the others (12.5 g ± 1.8 and 13.4 g ± 2.0). AST and LDH levels were higher in C-UW group (P<0.05). In the older group, there was a difference between UW and FBP preserved livers related to LDH, but not to AST. Thiobarbituric acid reactive substances were superior in control group than in the older one (P = 0.001). Catalase activity was similar between these groups (P = 0.11), but it was superior in UW preserved animals (P = 0.02). CONCLUSION: Livers from older rats were similar to the controls regarding cold ischemia injury in FBP group. Surprisingly, with UW solution there was less cold ischemia injury in the elderly group. When comparing both solutions, FBP provided significantly more protection than UW in the controls. There was a trend to FBP to being better than UW in the elderly group. Further studies with liver from older donors and ischemia and reperfusion are needed.

CONTEXT: Croton cajucara Benth is a plant found in Amazonia, Brazil and the bark and leaf infusions of this plant have been popularly used to treat diabetes and hepatic disorders. OBJECTIVES: This study investigated effects hepatics alterations and genotoxic and antidiabetic effect of Croton cajucara Benth bark extracts treatment in streptozotocin-induced diabetic rats. METHODS: Male Wistar rats were divided into six groups: control rats; control rats treated with Croton cajucara Benth extract during 5 and 20 days; diabetic rats, and diabetic rats treated with Croton cajucara Benth during 5 and 20 days. Diabetes was induced by a single intraperitoneal injection of streptozotocin (70 mg/kg). Eight weeks later we measured glucose, triglyceride, cholesterol and hepatic transaminases on blood. The bone marrow micronucleus assay was used to assess the genotoxic activity of Croton cajucara Benth. RESULTS: Treatment with aqueous extrat of Croton cajucara was able to significantly reduce levels of triglycerides in diabetic animals, however, did not modify significantly the levels of glucose and cholesterol in these animals. There was no significant elevation in liver transaminases in the control group treated with Croton cajucara Benth, as there was no genotoxic effect of treatment in this model. Our results did not show a significant effect on glucose and cholesterol reduction, the treatment was able to significantly reduce triclycerides plasmatic level. There was no significant alterations on hepatic transferase in the animals from the control group treated with Croton cajucara Benth. It was observed no genotoxic effect of the treatment in the model studied. CONCLUSION: In this study Croton cajucara bark extract showed absence of hepatotoxicity in this animal model and presented a hypolipidemic activity, and could be used to reverse dyslipidemia associated with diabetes and to prevent the cardiovascular complications that are very prevalent in diabetic patients.

CONTEXT: Enteroaggregative Escherichia coli strains have been associated with persistent diarrhea in several developing countries. In vivo procedures with animal models as rat, rabbit and gnotobiotic piglets intestinal loops, in vitro assays with cellular lines like T84, Caco 2, HT29, HeLa e HEp-2 and in vitro organ culture with intestinal fragments have been applied to study these bacteria and their pathogenicity. OBJECTIVES: The present experimental research assessed the pathogenic interactions of three enteroaggregative Escherichia coli strains, using the in vitro organ culture, in order to observe and compare alterations in different regions of both, the ileal and the colonic mucosa. METHODS: This study applied intestinal fragments from terminal ileum and colon that were excised from pediatric and adult patients that underwent colonoscopic procedures. Tissue was fixed for transmission electron microscopic study. Each bacterium was tested with three intestinal fragments for each region. RESULTS: Enteroaggregative Escherichia coli strains colonized and provoked citotoxic effects in the ileal and colonic mucosa. Total or partial villi destruction, vacuolization of basal cytoplasm of the enterocytes, epithelium detachment, derangement of the structure and epithelial cell extrusion in ileal mucosa could explain the perpetuation of the diarrhea. Bacterial aggregates were seen in intestinal lumen associated with mucus and cellular debris and in the intercellular spaces of the destroyed epithelium, suggesting bacterial invasion that seemed to be secondary to the destruction of the tissue. CONCLUSIONS: Pathogenesis of persistent diarrhea should include alterations in the small bowel structures where the digestive-absorptive functions take place. In the colonic mucosa the inflammatory lesions could explain the occurrence of colitis.

O conhecimento sobre os efeitos indesejáveis do uso de fungicidas convencionais, associado à preocupação de órgãos reguladores e consumidores quanto à qualidade dos alimentos, tem estimulado a busca por novas alternativas para o controle de doenças de plantas. O objetivo deste trabalho foi avaliar, em laboratório e em campo, a atividade antifúngica de 16 óleos essenciais sobre Phakopsora euvitis , agente causal da ferrugem da videira. Foram utilizados os óleos essenciais de orégano, menta piperita, pimenta preta, nim, eucalipto globulus , citronela, canela, manjerona, alecrim, manjericão, camomila azul, cânfora branca, cravo, gengibre, tomilho branco e melaleuca. No laboratório foram realizados dois experimentos: no primeiro, foi avaliada a germinação de esporos, em meio ágar-água acrescido do óleo essencial, nas concentrações de 0,0, 0,1, 0,5, 1, 2 e 4% (v/v), e sobre este foi depositada a suspensão de esporos. No segundo experimento, aplicaram-se os óleos essenciais em folhas destacadas de videira, seguindo-se a inoculação dos esporos de P. euvitis , avaliando-se a germinação dos esporos. A partir dos dados obtidos foram calculadas as percentagens de inibições da germinação de esporos e a dose letal (DL90). Em plantas de videira no campo foi realizada uma aplicação dos óleos essenciais, na concentração de 1%, e avaliada a severidade da ferrugem, após 15 e 30 dias. As DL90 dos óleos essenciais variaram entre 0,39% (nim) e 3,9% (orégano). Os óleos essenciais mais eficientes sobre P. euvitis , em condições in vitro , foram os de camomila azul, citronela, eucalipto globulus , gengibre, nim e tomilho branco. Na aplicação realizada em campo todos os óleos essenciais reduziram significativamente a severidade da ferrugem, destacando-se os óleos essenciais de canela, citronela, nim e tomilho branco. Aspectos relacionados à fitotoxicidade necessitam ser elucidados. Os óleos essenciais avaliados apresentam potencial para serem utilizados no manejo da ferrugem da videira, contudo estudos complementares fazem-se necessários.

Produtos fitossanitários podem influenciar de maneira negativa a atratividade ou o comportamento de visitantes florais. Dessa forma, o presente trabalho teve como objetivo avaliar a influência desses produtos na diversidade de espécies e frequência de visitação floral nas culturas do pimentão e do tomateiro após a aplicação de produtos fitossanitários. Os produtos aplicados foram ciantraniliprole, ciantraniliprole + tiametoxam, tiametoxam, imidacloprido, ciantraniliprole + abamectina e diafentiurom. Os ensaios foram realizados em campo no período de junho a novembro de 2012. Foram avaliados o número total de espécies e o número de visitas de visitantes florais no período de 26 de junho de 2012 a 11 de setembro de 2012 para a cultura do tomateiro e de 25 de setembro de 2012 a 13 de novembro de 2012 para a cultura do pimentão. Para o cultivo do tomateiro, foi encontrado um total de 8 espécies de visitantes florais; para a cultura do pimentão, foram verificadas 27 espécies. Os produtos fitossanitários aplicados tanto na cultura do tomateiro quanto na do pimentão não reduziram o número de visitantes florais nem a frequência de visitação dos insetos, demonstrando seletividade ecológica.

As culturas oleaginosas vêm ganhando destaque devido ao seu potencial para a produção de biodiesel, como uma alternativa ao diesel. Portanto, neste trabalho objetivou-se avaliar a entomofauna, por meio de análises faunísticas, associada às seguintes culturas: nabo forrageiro, niger, crambe e girassol, no município de Dourados, estado de Mato Grosso do Sul, tendo em vista que esses cultivares possuem potencial para a produção de biodiesel. Os insetos foram coletados com o método de pano de coleta. As avaliações ocorreram em área experimental agrícola (latitude de 22º 14'S, longitude de 54º 49'W). Foi observado um total de 7.406 espécimes nas quatro culturas, correspondendo a 3.569 espécimes no nabo forrageiro, o qual apresentou uma riqueza de insetos com 18 espécies; 2.275 espécimes no niger, com uma riqueza de 17 espécies; 908 espécimes no crambe com uma riqueza de 18 espécies; e 654 espécimes no girassol com uma riqueza de 18 espécies. Com relação ao estudo da diversidade de espécies medida nas quatro culturas analisadas, o índice de Shannon-Wiener (H') indicou baixa diversidade apenas na cultura do nabo forrageiro, uma vez que o maior valor ocorreu na cultura do girassol (22,935), seguida por crambe (17,487), niger (15,364) e nabo forrageiro (0,5716). A cultura que apresentou o maior índice de equitabilidade foi a do girassol (0,7935), seguida por crambe (0,6047), niger (0,5423) e nabo forrageiro (0,1978), indicando uma semelhança de composição faunística entre girassol, crambe e niger.

The aims of this study were to assess in vitro if bovine oocytes and oviductal epithelial cells from slaughterhouses for in vitro fertilization use may be infected with bovine herpesvirus 1; to analyze whether the treatment with trypsin according to the International Embryo Transfer Society guideline is efficient to inactivate the bovine herpesvirus 1; to morphologically study the virus-oocyte interaction through optical microscopy. In this study, Madin Darby Bovine Kidney (MDBK) cells that were co-cultured with oocytes matured in vitro and exposed to bovine herpesvirus 1 showed a cytopathic effect. The nested polymerase chain reaction for the supernatant was positive for the bovine herpesvirus 1, thus suggesting that the cytopathic effect observed in the MDBK monolayer was seen due to virus replication and not because of any culture toxicity. It was also observed cytopathic effect and positive nested polymerase chain reaction in MDBK cells co-cultured with in vitro maturated oocytes free of virus, but that were co-cultured in uterine epithelial cells pre-infected with bovine herpesvirus 1 and washed or not with trypsin, demonstrating an oocyte contamination by the virus. When trypsin-washing efficacy was evaluated, we could observe that the trypsin treatment was not able to eliminate the bovine herpesvirus 1 of the oocytes, and it was not observed any morphological difference in the infected oocytes.

Caseous lymphadenitis is a chronic infectious disease caused by Corynebacterium pseudotuberculosis , which is a bacterium responsible for a great number of economic losses on goat and sheep production. It is characterized by the formation of abscesses in superficial lymph nodes and in internal organs and lymph nodes. This study aimed at determining the agreement between microbiological culture and PCR in the identification of C. pseudotuberculosis , in samples collected from animals in slaughterhouses and in animals that presented lymph node enlargement in field conditions. From the 202 samples analyzed through microbiological culture, 113 (56%) were positive for Corynebacterium sp.; from these positive samples, 38 (34%) were identified as C. pseudotuberculosis by microbiological culture. From the amount of samples, 110 (54%) were positive and 92 (46%) were negative in the PCR. Kappa index (0.193) presented a weak agreement between PCR and microbiological culture. We concluded that molecular diagnosis (PCR) in clinical samples proved to be more efficient, reproducible, and faster than the microbiological culture, both on clinical samples analyses and in the confirmation of C. pseudotuberculosis in colonies that were classified by Corynebacterium genus. Thus, the present study demonstrated the importance of PCR to confirm C. pseudotuberculosis diagnosis, and the best contribution for the epidemiological surveillance of the disease in sheep.

RESUMO: A vacinação contra febre aftosa é ferramenta eficaz para erradicação e prevenção da doença, contudo, a presença de proteínas não capsidais (PNC) na vacina, produzidas durante a multiplicação viral, tem sido o principal problema, visto que sua presença dificulta as ações de vigilância, cuja busca se baseia na detecção de anticorpos contra essas proteínas para diferenciar animal vacinado de infectado. Por esse motivo, o Ministério da Agricultura, Pecuária e Abastecimento publicou em 2008 a Instrução Normativa nº 50 (IN 50), que incluiu no controle da qualidade da vacina a pesquisa de anticorpos contra as PNC, para avaliar a retirada dessas proteínas. Considerando a interferência vacinal, objetivou-se avaliar a frequência de bovinos reagentes às PNC, analisada pelo sistema I-ELISA 3ABC/EITB, no Laboratório de Viroses de Bovídeos do Instituto Biológico de São Paulo, no período de 2002 a 2012. Dos 34.705 bovinos examinados, observou-se que a proporção de reagentes às PNC aumentou com a idade, evidenciando aumento da frequência de reagentes em animais que receberam maior número de vacinações, indicando interferência da vacinação na resposta às proteínas. Quando comparados antes e após a publicação da IN 50, observa-se diminuição da reatividade, com redução, em 2010, de quase a metade em relação a 2007, e ainda maior quando comparado com o período 2002 a 2006. Isso demonstra a efetividade da purificação da vacina em resposta ao cumprimento da IN 50, embora permaneça alguma sororreatividade em bovinos com múltiplas vacinações. O sistema I-ELISA 3ABC/EITB demonstrou ser uma ótima ferramenta para impedir a movimentação de possíveis portadores do vírus oriundos de rebanhos vacinados, desde que seja considerado todo o contexto sanitário e epidemiológico.

RESUMO O objetivo do trabalho foi avaliar a diminuição da replicação viral (BoHV-1 Colorado) em embriões murinos após tratamento do extrato etanólico da casca de Punica granatum (EEPg). Camundongos fêmeas Swiss com idade entre 6 e 8 semanas foram superovuladas com 0,2 mL a 5 UI de hormônios (eCG e hCG), e acasaladas com machos da mesma idade. Após 18 horas, as fêmeas sofreram eutanásia em câmara de CO2 e, através de abertura no peritônio, os zigotos foram coletados e lavados com solução de pronase 0,5%.Os zigotos foram divididos em quatro grupos: G1 (controle), G2 (expostos aos vírus BoHV-1 Colorado a 108 TCID50/mL), G3 (expostos ao EEPg) e G4 (expostos aos vírus e ao EEPg). Os grupos foram mantidos a 37,5ºC em meio TCM199 (100µL) com 10% de soro fetal bovino em estufa a 5% de CO2 e 95% de umidade. Após 24 h, analisamos a taxa de clivagem (teste exato de Fisher; p<0,05), a morfologia (por microscopia óptica), a nested-PCR e a titulação dos embriões em cocultura com células MDBK após mais 72 h do tratamento (teste de Mann-Whitney; p<0,05) e microscopia eletrônica de transmissão (ME). Os embriões murinos tratados com EEPg apresentaram resultados satisfatórios: sem alterações morfológicas, taxa de clivagem semelhante ao controle e, apesar da detecção da presença do vírus pela nested-PCR e ME, houve diminuição do título viral após tratamentos com esse extrato, o que sugere interferência desse tratamento no ciclo viral do BoHV-1 Colorado sem alterar o desenvolvimento dos embriões.

RESUMEN Las especies Duddingtonia flagrans y Monacrosporium thaumasium son micro-hongos considerados promisorios agentes del control biológico de parásitos. Bajo condiciones adversas como la falta de nutrientes, estos hongos producen esporas capaces de sobrevivieren después de pasar por el tracto gastrointestinal de los animales. La formación de estas estructuras es una característica deseable ya que promueve la sobrevivencia y la diseminación de los hongos para propósitos de biocontrol. El objetivo de este estudio fue evaluar la producción de esporas de dos especies de hongos nematófagos D. flagrans (aislados AC001 y CG722) y M. thaumasium (NF34A). Estos fueron cultivados en los subproductos agroindustriales, que tenían el intento de identificar el mejor medio para uso en programas de biocontrol de nematodos. Diferentes volúmenes (10, 15 y 20 mL) de masa micelial fueron utilizados como inóculos iniciales y adicionados a 100 gramos de medios de crecimientos sólidos (sémola de arroz - QA; sémola de maíz - QM; bagazo de caña - BC; paja de arroz - PA y cascara de café - CC) y mantenidos a 25°C en la obscuridad para evaluar la producción de esporas. Los aislados AC001 y CG722 mostraron las mejores producciones en el medio de la QA (p<0,05). El volumen de 20 mL de masa micelial utilizado como inóculo inicial proporcionó una mayor recuperación de esporas. El aislado NF34A presentó una baja o nula producción de estructuras reproductivas en los diferentes volúmenes y medios de crecimientos utilizados. La mejor producción de esporas se obtuvo utilizando subproductos de la agroindustria con mayor densidad proteica y energética.