External contamination of broilers by Campylobacter spp. increases from the farm to the slaughterhouse.

1. In this study, classical and molecular microbiological methods for detection and quantification of Campylobacter spp. were used to estimate their prevalence in faecal samples and skin swabs collected from 31 broiler flocks (20 farms) in Portugal, and measure the impact of transport-related factors on the expected rising excretion rates from the farm to the slaughterhouse. 2. Data on husbandry practices and transport conditions were gathered, including time in transit, distance travelled or ante-mortem plant-holding time. 3. A generalised linear mixed model was used to evaluate the significance of a potential post-transport rise in Campylobacter spp. counts and to assess risk determinants. 4. At least one flock tested positive for Campylobacter spp. in 80% of the sampled farms. At the slaughterhouse, Campylobacter spp. were detected in all faecal samples, C. jejuni being the most commonly isolated. 5. A post-transport rise of Campylobacter spp. counts from skin swabs was observed using classical microbiological methods (from a mean of 1.43 to 2.40 log10 CFU/cm2) and molecular techniques (from a mean of 2.64 to 3.31 log10 genome copies/cm2). 6. None of the husbandry practices or transport-related factors were found to be associated with Campylobacter spp. counts. 7. This study highlights the need for more research to better understand the multi-factorial nature of Campylobacter spp., a public health threat that was found to be highly prevalent in a sample of Portuguese poultry farms.

Taxas de perda gestacional até 60 dias são afetadas por características cíclicas da égua receptora de embrião Mangalarga Marchador / Rates of pregnancy loss up to 60 days are affected by cyclic characteristics of the Mangalarga Marchador recipient mare

Foram avaliadas taxas de gestação aos 15 dias e perda gestacional entre 15 e 60 dias em 430 transferências de embrião (TE) em éguas Mangalarga Marchador. Diagnósticos de gestação foram realizados entre 15 e 60 dias após TE. Para avaliar os efeitos da duração da fase folicular da receptora, foram formados três grupos: até três dias (<3d); quatro a seis dias (4-6d); sete ou mais dias (>7d). Para avaliar os efeitos do tamanho do folículo pré-ovulatório da receptora, foram formados outros três grupos: menor ou igual a 35mm (Ø<35); maior que 35 e menor ou igual a 45mm (35<Ø<45); maior que 45mm (Ø>45). Os grupos foram comparados pelo teste qui-quadrado (P<0,05). Quanto à duração da fase folicular, as taxas de gestação foram semelhantes (<3d - 83,1%; 4-6d - 86,4%; >d - 86,0%), e a perda gestacional maior em >7d (24,4%) que em <3d(12,0%) e 4-6d (13,3%), estas semelhantes entre si. Quanto ao tamanho folicular, as taxas de gestação foram semelhantes (Ø<35 - 86,4%; 35<Ø<45 - 86,5%; Ø>45 - 81,9%), assim como as de perda gestacional (Ø<35 - 13,2%; 35<Ø<45 - 18,1%; Ø>45 - 10,5%). Razões para a maior perda gestacional no grupo >7dnão foram esclarecidas, mas conclui-se que a duração da fase folicular pode ser fator de escolha de receptoras.

Pregnancy rates were evaluated at 15 days and pregnancy loss between 15 and 60 days on 430 embryo transfers (ET) in Mangalarga Marchador mares. Pregnancy diagnosis was performed between 15 and 60 days after ET. To evaluate the effects of the duration of the follicular phase of the recipient mare, three groups were formed: up to 3 days (<3d); 4 to 6 days (4-6d); 7 or more days (>7d). To evaluate the effects of the size of the pre-ovulatory follicle of the recipient mare, three other groups were formed: below or equal to 35mm (Ø<35); greater than 35 and below or equal to 45mm (35<Ø<45); greater than 45mm (Ø>45). The groups were compared by Chi-square test (P<0.05). Regarding the duration of the follicular phase, pregnancy rates were similar (<3d - 83.1%; 4-6d - 86.4%; >7d - 86.0%), and greater pregnancy loss in >7d (24.4%) than in <3d (12.0%) and 4-6d (13.3%), which were similar. Regarding the follicle size, pregnancy rates (Ø<35 - 86.4%; 35<Ø<45 - 86.5%; Ø>45 - 81.9%) and pregnancy loss (Ø<35 - 13.2%; 35<Ø<45 - 18.1%; Ø>45 - 10.5%) were similar. Reasons for the greatest pregnancy loss in the >7d group have not been elucidated, but we conclude that the duration of the follicular phase may be a factor for choosing recipient mares.

Risk factors for hepatitis C virus infection in former Brazilian soccer players.

We evaluated the occurrence of hepatitis C virus (HCV) infection in 97 former soccer players who played in Recife, Brazil in the 1960s and 1970s, and analysed the risk factors for infection, such as history of transfusions, surgery, tattoos, piercings, and the use of illicit drugs or injectable vitamin complexes. Immunochromatographic testing was performed to detect anti-HCV antibodies. All former soccer players were men (mean age 59·2 years), of whom 62 (64%) and 35 (36%) were classified as amateurs and professionals, respectively. Seven (7·2%) tested positive for anti-HCV antibodies; three (4·8%) were amateurs, and four (11·4%) were professionals. In univariate analysis, transfusion, surgery, and use of injectable vitamin complexes were associated with HCV infection, while in multivariate analysis, only the use of injectable vitamin complexes was related (P=0·0005). We observed a high frequency of HCV infection in former soccer players, especially in professionals who used injectable vitamin complexes.

Schistosomiasis mansoni: ultrasound-evaluated hepatic fibrosis and serum concentrations of hyaluronic acid.

Schistosomiasis mansoni is a fibrogenic liver disease that constitutes a major health problem in north-eastern Brazil. Although one common manifestation of the disease, periportal fibrosis (PPF), can be assessed by ultrasonography by well-trained physicians, the necessary equipment and personnel are not always readily available. Serum markers, including hyaluronic acid (HA), have been used as alternative means of measuring fibrosis. Recently serum concentrations of HA have been evaluated in 77 Brazilians (61 cases of schistosomiasis mansoni and 16 healthy controls) and compared against the ultrasound-evaluated PPF in the same subjects. The HA was measured using a non-competitive fluorescence-based assay, while the PPF was explored using a portable ultrasound scanner (SSD-500; Aloka, Tokyo) and graded, as patterns A-F, according to the World Health Organization's 'Niamey protocol'. In general, the serum concentrations of HA were found to be positively correlated with the severity of the PPF. The mean concentration of HA in the sera of the 16 controls was significantly lower than that recorded in the schistosomiasis cases who showed PPF of patterns D or E (P<0·001 for each). The cases who showed pattern-C PPF also had significantly less HA in their sera than the cases with PPF of patterns D or E (P<0·001 for each), and the cases with pattern-D fibrosis had significantly lower HA concentrations in their sera than the cases with PPF of pattern E (P<0·001). In an analysis based on a receiver-operating-characteristic (ROC) curve, an HA concentration of 20·2 µg/litre of serum was identified as a threshold that could be used to distinguish moderate cases of PPF (i.e. patterns C or D) from the more advanced cases (i.e. patterns E or F), with a sensitivity of 60% and specificity of 65%. In conclusion, it appears that serum concentrations of hyaluronic acid could be used as markers for periportal fibrosis in patients with schistosomiasis mansoni.

Effects of timing of induced luteolysis in embryo donor mares on reproductive performance and pregnancy rate in recipient mares.

The objective was to evaluate the effects of giving prostaglandin F2(α) (PGF) to donor mares 48 h prior to embryo collection. Non-lactating donor mares (n = 20 estrous cycles in 10 mares), ranging from 2.5 to 10 y of age and 400 to 500 kg of body weight were used from September 2004 to February 2005 in the southern hemisphere (Brazil). Donor mares were randomly assigned in a cross-over design study. During a Treated cycle, 7.5 mg PGF was given 48 h prior to embryo collection, whereas in the Control cycle, 7.5 mg PGF was given at embryo collection. In Treated Cycles, serum progesterone concentrations decreased between the day of PGF treatment and the day of embryo collection (13.9 ± 5.4 and 0.5 ± 0.3 ng/mL, respectively; P < 0.05). In Treated versus Control cycles, the interovulatory interval was shorter (14.9 ± 0.9 vs 17.5 ± 1.1 d, P < 0.05). However, there was no significant difference between these groups for the interval from PGF to ovulation (average, 9.8 d), embryo recovery rate (average, 75%), embryo quality, uterine protein concentration, and pregnancy rate in recipient mares (average, 87% at 15 d after ovulation, with no pregnancy loss detected by 60 d). In conclusion, giving donor mares PGF 48 h prior to embryo collection reduced the average interovulatory interval by approximately 2.5 d, thereby potentially increasing the numbers of embryos that could be collected during a breeding season, with no deleterious effects on embryo recovery rate, embryo quality, or pregnancy rate in recipient mares.

Hepatitis C virus infection diagnosis using metabonomics.

Metabonomics based on nuclear magnetic resonance (NMR) can reveal the profile of endogenous metabolites of low molecular weight in biofluids related to disease. The profile is identified a 'metabolic fingerprint' like from the pathological process, why this metabonomics has been used as a diagnostic method. The aim of the present study was to apply metabonomics to identify patients infected with the hepatitis C virus (HCV) through an analysis of ¹H NMR spectra of urine samples associated with multivariate statistical methods. A pilot study was carried out for the diagnostic test evaluation, involving two groups: (i) 34 patients positive for anti-HCV and HCV-RNA and negative for anti-HBc (disease group); and (ii) 32 individuals positive for anti-HBc and negative for HBsAg and anti-HCV. The urine samples were analyzed through ¹H NMR, applying principal component analysis and discriminant analysis for classification. The metabonomics model was capable of identifying 32 of the 34 patients in the disease group as positive and 31 of the 32 individuals in the control group as negative, demonstrating 94% sensitivity and specificity of 97% as well as positive and negative predictive values of 97% and 94%, respectively, and 95% accuracy (P < 0.001). In conclusion, the metabonomics model based on ¹H NMR spectra of urine samples in this preliminary study discriminated patients with HCV infection with high sensitivity and specificity, thereby demonstrating this model to be a potential tool for use in medical practice in the near future.

Use of a low dose of equine purified FSH to induce multiple ovulations in mares.

The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19 degrees 45'45'S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post-ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35-mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35- to 40-mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively--one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low-dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.

Determination of the cut-off value of serum alanine aminotransferase in patients undergoing hemodialysis, to identify biochemical activity in patients with hepatitis C viremia.

BACKGROUND: Elevated liver enzymes are infrequent in patients with hepatitis C virus (HCV) infection undergoing chronic hemodialysis (HD), suggesting that the alanine aminotransferase (ALT) is a poor predictor of hepatocellular damage in this population. OBJECTIVE: To establish a more appropriate cut-off value of ALT to identify biochemical activity due to HCV infection in HD patients. STUDY DESIGN: A total of 217 patients, with an average age of 51.2 years, were evaluated between January and October 2002; 130 were males (60%). Serum ALT was measured by a kinetic method in five consecutive monthly blood samples, from which an average was obtained and divided by the upper limit of normal (ULN). HCV antibodies were determined using an enzyme immunoassay, the serum HCV-RNA by nested-PCR and HCV genotype by hybridization of the amplified sequence from the 5'-non-coding region. The cut-off value of ALT was obtained from a ROC curve. RESULTS: Within the 217 patients, 18 (8.3%) were anti-HCV-positive, 17 (7.8%) of whom were also HCV-RNA-positive. Genotype distribution was: 1a=47%; 1b=18%; 3a=35%. Mean ALT/ULN (0.77+/-0.57) of the 18 anti-HCV-positive cases was higher (p<0.001) than the negative group (0.38+/-0.23). The mean ALT/ULN (0.81+/-0.57) of the 17 HCV-RNA-positive cases was also higher (p<0.0001) than the negative cases (0.37+/-0.23). The cut-off value of ALT to distinguish the anti-HCV-positive from negative patients was 0.50% or 50% of the ULN (sensitivity=67%; specificity=83%). According to the HCV-RNA, the cut-off value of ALT was 0.45% or 45% of the ULN (sensitivity=71%; specificity=80%). CONCLUSION: Reducing the cut-off of ALT by half, enables a better identification of biochemical activity in patients with HCV infection on chronic HD.

[Splenectomy plus left gastric vein ligature and devascularization of the great curvature of the stomach in the treatment of hepatosplenic schistosomiasis. Postoperative endoscopic sclerosis is necessary?]. / Esplenectomia com ligadura da veia gástrica esquerda e desvascularização da grande curvatura do estômago no tratamento da esquistossomose hepatoesplênica. E necessária a escleroterapia endoscópica pós-operatória?

OBJECTIVE: With the intention of evaluating the effectiveness and the maintenance of the postoperative endoscopic sclerosis as routine, in association to splenectomy with left gastric vein ligature and devascularization of the great curvature of the stomach, the present study was accomplished. METHOD: Between 1992 and 1998, 131 patient were operated in the General Division of the "Hospital das Clínicas" (Federal University of Pernambuco, Recife, PE, Brazil). The medium follow-up was 30 months. All patients were requested to come back to the clinic for accomplishment of clinical and laboratory control. Of the 111 patients that came back to the clinic, 80 patients had a digestive endoscopy done. Of these 80 patients, 36 followed the recommendation and underwent to a postoperative endoscopic sclerosis program (group 1), while 44 did not accomplish postoperative endoscopic sclerosis (group 2). RESULTS: Regarding the eradication of the esophagus varices, the authors found a statistical difference between the groups (52.7% of the group 1 vs. 18.2% of the group 2). Other analyzed items (mortality, rebleeding rate, thrombosis of the portal vein, gastric varices and degree of periportal fibrosis) statistical relevance was not observed. CONCLUSION: The association of the postoperative endoscopic sclerosis to the splenectomy with left gastric vein ligature and devascularization of the great curvature of the stomach, in the treatment of schistosomotic portal hypertension with digestive hemorrhage antecedent, should be maintained.

Evaluation of an enzyme immunoassay for hepatitis C virus antibody detection using a recombinant protein derived from the core region of hepatitis C virus genome

This study was undertaken to evaluate an enzyme immunoassay (EIA) for hepatitis C virus antibody detection (anti-HCV), using just one antigen. Anti-HCV EIA was designed to detect anti-HCV IgG using on the solid-phase a recombinant C22 antigen localized at the N-terminal end of the core region of HCV genome, produced by BioMérieux. The serum samples diluted in phosphate buffer saline were added to wells coated with the C22, and incubated. After washings, the wells were loaded with conjugated anti-IgG, and read in a microtiter plate reader (492 nm). Serum samples of 145 patients were divided in two groups: a control group of 39 patients with non-C hepatitis (10 acute hepatitis A, 10 acute hepatitis B, 9 chronic hepatitis B, and 10 autoimmune hepatitis) and a study group consisting of 106 patients with chronic HCV hepatitis. In the study group all patients had anti-HCV detected by a commercially available EIA (Abbott(r)), specific for HCV structural and nonstructural polypeptides, alanine aminotransferase elevation or positive serum HCV-RNA detected by nested-PCR. They also had a liver biopsy compatible with chronic hepatitis. The test was positive in 101 of the 106 (95 percent) sera from patients in the study group and negative in 38 of the 39 (97 percent) sera from those in the control group, showing an accuracy of 96 percent. According to these results, our EIA could be used to detect anti-HCV in the serum of patients infected with hepatitis C virus.

Evaluation of an enzyme immunoassay for hepatitis C virus antibody detection using a recombinant protein derived from the core region of hepatitis C virus genome.

This study was undertaken to evaluate an enzyme immunoassay (EIA) for hepatitis C virus antibody detection (anti-HCV), using just one antigen. Anti-HCV EIA was designed to detect anti-HCV IgG using on the solid-phase a recombinant C22 antigen localized at the N-terminal end of the core region of HCV genome, produced by BioMérieux. The serum samples diluted in phosphate buffer saline were added to wells coated with the C22, and incubated. After washings, the wells were loaded with conjugated anti-IgG, and read in a microtiter plate reader (492 nm). Serum samples of 145 patients were divided in two groups: a control group of 39 patients with non-C hepatitis (10 acute hepatitis A, 10 acute hepatitis B, 9 chronic hepatitis B, and 10 autoimmune hepatitis) and a study group consisting of 106 patients with chronic HCV hepatitis. In the study group all patients had anti-HCV detected by a commercially available EIA (Abbott), specific for HCV structural and nonstructural polypeptides, alanine aminotransferase elevation or positive serum HCV-RNA detected by nested-PCR. They also had a liver biopsy compatible with chronic hepatitis. The test was positive in 101 of the 106 (95%) sera from patients in the study group and negative in 38 of the 39 (97%) sera from those in the control group, showing an accuracy of 96%. According to these results, our EIA could be used to detect anti-HCV in the serum of patients infected with hepatitis C virus.

A prospective and randomized study using ribavirin as monotherapy for the treatment of naïve patients with chronic hepatitis C.

In order to evaluate the response to ribavirin in previously untreated patients with chronic hepatitic C, 39 patients were selected for a double-blind prospective and randomized trial, and divided into two groups: ribavirin-group (19 patients) and placebo-group (20 patients). Ribavirin was administered orally for 24 weeks (600 mg/day, followed by 1,000 mg/day and 1,200 mg/day each one for 8 weeks). After 3 months of drug administration, the patients were evaluated by measuring biochemical, virologic and histologic responses. After this phase, ribavirin was offered to the patients who had received placebo (second phase). The results showed that the patients who received ribavirin showed a higher reduction in serum alanine aminotransferase (ALT) activity than patients in the placebo group. Among the patients in the ribavirin-group, a complete biochemical response (ALT levels normalized) was observed in 3 patients (16%), and a partial response (reduction greater than 50% of the initial value of ALT activity) in 4 (21%). In the 20 patients in the placebo group, only 1 showed a partial response (5%). In the second phase of the study, among 16 patients who received ribavirin, 4 (25%) showed a complete and 5 (31%) a partial biochemical response. HCV-RNA did not become negative in any patient during the two phases. A reduction in the score of portal and lobular activity was observed in patients who received ribavirin, but statistical analysis did not identify differences. This study showed that ribavirin alone induces a biochemical response (ALT reduction) in some patients with chronic hepatitis C, which may be associated with a reduction in hepatic inflammatory activity reduction, but the changes are not sufficient to recommend initial monotherapy with ribavirin.

[Glomerulonephritis associated with hepatitis c virus infection]. / Glomerulonefrite associada à infecção pelo vírus da hepatite C.

We report 4 patients with glomerulonephritis (GN) associated with hepatitis C virus (HCV) infection seen between August 1993 and July 1996. Two of them were male and median age was 41 years. Anti-HCV was detected by enzyme-immunoassay and HCV-RNA by PCR. Serum cryoglobulins, 24-hour proteinuria, and erythrocyte dismorphism were also determined. Viremia, cryoglobulinemia, hematuria and proteinuria were observed in all patients. Liver biopsies revealed inflammatory activity in 3 cases, and renal biopsies revealed membranoproliferative glomerulonephritis in 3 patients and mesangial proliferative glomerulonephritis in 1 patient. Two patients are on specific therapy for HCV infection (IFN in combination with ribavirin) and have presented clinical and laboratory improvement. The occurrence of active liver disease and viremia concurrent with urinary alterations suggests viral involvement in renal disease, a conclusion supported by the by improvement of urinary alterations observed after treatment for HCV. We conclude that the search for viral markers in patients with GN is important since their detection could change the therapeutic approach.

Prevalence of hepatitis C virus antibodies in primary glomerulonephritis in Brazil.

The association between hepatitis B virus and membranous glomerulonephritis and membranoproliferative glomerulonephritis (MPGN) was first described in 1971. Recently, a similar association between hepatitis C virus (HCV) and glomerulonephritis (GN) has been reported. We investigated the prevalence of hepatitis C serum antibodies (anti-HCV) in patients with primary GN followed up at our Nephrology Outpatient Clinic between March 1993 and November 1995. The diagnosis of primary GN was established after excluding the presence of connective tissue disease, diabetes, infectious disease, and malignancy. Anti-HCV antibodies were detected by a second-generation enzyme immunosorbent assay and HCV RNA by polymerase chain reaction. Of 81 patients with primary GN, 24 had membranous glomerulonephritis, 17 MPGN, 15 minimal-change disease, 12 focal-segmental glomerulosclerosis, 9 diffuse proliferative GN, and 4 IgA nephropathy. Anti-HCV were detected in 2 cases (2.5%), both were HCV RNA positive and had a polyclonal mixed cryoglobulinemia (IgM-IgG). These 2 cases both came from the group of 17 patients with MPGN. Biochemical investigation in these patients revealed persistent elevation of serum aminotransferase activity, and a liver biopsy specimen in 1 of them showed evidence of chronic active hepatitis. We conclude that in our setting the prevalence of anti-HCV among patients with primary GN is low, being higher (11.8%) only if we consider the patients with MPGN as the reference group. Further studies are necessary to clarify this association and to determine appropriate therapy for these patients.

Autoantibodies before, during and after administration of recombinant interferon-alpha for chronic viral hepatitis.

This study was undertaken to investigate the presence of autoantibodies in patients with chronic viral hepatitis B and C, before, during and after interferon-alpha (IFN-alpha) therapy and to study their relation to dose and type of IFN-alpha and response to treatment. Fifty patients with chronic hepatitis were divided in two groups, a control-group of 21 patients (10 type B and 11 type C) who were followed for 6 months without treatment and an IFN-group consisting of 29 patients (8 type B and 21 type C) who received IFN therapy for 6 months. Serum samples were tested for a range of antibodies at the start of the study, during therapy and at the end of the 6 month period. Antibodies tested for included: antinuclear, smooth muscle, antimitochondrial, parietal cell and thyroid microsomal. Four (8%) of the total patient group had autoantibodies at the beginning of the study (two in each group). During the follow-up period no patient in the control group developed antibodies compared with 3 (11%) patients in the treatment group. Autoantibodies developed in patients treated with higher doses of IFN and were found in those patients who tended to show a poor response to IFN-therapy. Further studies are needed to establish the relationship between poor response to IFN-alpha and development of autoantibodies.