Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection.

Validation of two real-time PCRs targeting the PE-PGRS 20 gene and the region of difference 4 for the characterization of Mycobacterium bovis isolates.

This study aimed to develop and validate real-time PCR for the diagnosis of Mycobacterium bovis isolates. Two hundred and seventy-four M. bovis isolates and 156 M. tuberculosis isolates were tested. Both qPCRs amplified all of the 274 M. bovis samples, but none of the 156 M. tuberculosis samples. The qPCR for PE-PGRS 20 had 91% efficiency and a detection limit of 0.32 ng (sensitivity and specificity for qPCR "Mbovis.100" were 99.64 and 100%, respectively). The qPCR for RD4 had 100% efficiency, and a detection limit of 4 pg (diagnostic sensitivity and specificity were 100 and 100%. The qPCR tests were performed using 4 extraction sets, 3 qPCR kits, and with a range of equipment; yet, all combinations produced similar results in a diagnostic test, demonstrating the robustness of this method. The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.

Serological and molecular investigation of the prevalence of Aujeszky's disease in feral swine (Sus scrofa) in the subregions of the Pantanal wetland, Brazil.

The feral swine (FS) originated from the domestic pig and is present throughout the Brazilian wetland plain (the Pantanal). Aujeszky's disease (AD) was first serologically confirmed in the state of Mato Grosso do Sul (MS) in 2001; however, there was no viral confirmation. The aim of this study was to investigate antibodies against-SuHV-1 in the sera of feral swine in the studied areas, detect SuHV-1 through PCR and classify the viral genome. Among the 218 animals sampled, 186 were analyzed by ELISA, resulting in 88 (47.3%) reactive samples. In the serum neutralization test (SN), 57/179 (31.8%) samples presented antibodies against the AD virus (SuHV-1). By nested PCR, 104 DNA samples were extracted for analysis and confirmed with amplification of a fragment of glycoprotein B (gB) in five samples. The SuHV-1 was detected in 12 samples by using primers for glycoprotein E (gE) and viral genome was classified as Type I by ul44 partial sequencing. The amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows indicate that the vertical transmission contribute to maintain SuHV-1 in a free-living feral swine population. The origin of AD in the feral swine populations of the Pantanal is unknown, however, the determination of viral latency, the vertical transmission of the antigen by the amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows and genome typing contribute to the elucidation of the epidemiology of this disease in the wetlands of MS, Brazil.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r £ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry.

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r ≤ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Pseudorabies virus can be classified into five genotypes using partial sequences of UL44.

Suid herpesvirus 1 (SuHV-1) is the causative agent of pseudorabies (PR), a disease of great importance due to the huge losses it causes in the swine industry. The aim of this study was to determine a method for genotyping SuHV-1 based on partial sequences of the gene coding for glycoprotein C (gC) and to elucidate the possible reasons for the variability of this region. A total of 109 gCsequences collected from GenBank were divided into five major groups after reconstruction of a phylogenetic tree by Bayesian inference. The analysis showed that a portion of gC (approximately 671 bp) is under selective pressure at various points that coincide with regions of protein disorder. It was also possible to divide SuHV-1 into five genotypes that evolved under different selective pressures. These genotypes are not specific to countries or continents, perhaps due to multiple introduction events related to the importation of swine.

A comparative analysis of envelope and tegument proteins of suid herpesvirus 1, bovine herpesvirus 1 and bovine herpesvirus 5.

The aim of this study was to analyze the disordered regions (DRs) in the envelope and tegument proteins of three closely related herpesviruses: bovine herpesvirus 1, bovine herpesvirus 5 and suid herpesvirus 1. Tegument proteins showed a greater percentage of DRs than the envelope proteins did. Regions of disorder were found in the less conserved portions of the proteins, N-terminal and C-terminal regions, and, in a few cases, functionally important sites. The presence of DRs is an important factor in the evolution of viruses, representing points where positive pressure led to structural changes.

Effect of 10% formalin on radiographic optical density of bone specimens.

OBJECTIVES: The aim of this work was to determine whether the fixation process with 10% formalin modifies the radiographic optical density of bone. METHODS: Eight rabbit tibiae were placed in separate containers with one of the following fixative solutions: 10% formalin (n = 3), 10% phosphate-buffered formalin (n = 3) or 10% calcium carbonate-buffered formalin (n = 2). Radiographs were obtained at 5 different times: before fixation (time zero), then 1 day, 15 days, 30 days and 90 days following immersion in the fixative solution. Radiographs were scanned and optical density was determined using ImageTool software. RESULTS: There was no significant difference in radiographic optical density among the specimens fixed in 10% formalin (146.60+/-32.44), 10% phosphate-buffered formalin (149.84+/-32.43) and 10% calcium carbonate-buffered formalin (146.61+/-35.92). Regardless of buffering, optical density at time zero was significantly higher than that at 15 days, 30 days and 90 days. However, while optical density at 1 day was significantly higher than that at 30 days and 90 days, it did not differ from that at 15 days. There was also no significant difference in density between 30 days and 90 days (ANOVA, Tukey, at 5% level of significance). CONCLUSION: The radiographic optical density of specimens stored in 10% formalin diminishes with time, irrespective of buffering, which suggests the occurrence of bone demineralization.

[Congenital hypothyroidism screening: seasonal variations of TSH levels] / Variação sazonal nos níveis deTSH em testes de rastreamento para hipotireoidismo congênito.

OBJECTIVE: To evaluate the seasonal differences in newborn TSH levels. METHOD: A restrospective study was performed using normal blood TSH measurements collected in a neonatal screening program for congenital hypothyroidism, during the extremes of temperature (summer/winter) in order to verify the seasonal influence on that exam. Blood samples were collected from baby feet on filter paper in health care public institutions in Rio de Janeiro city and TSH were analyzed by immunofluorimetric method. For this study we considered samples with TSH levels less then 20 micro U/ml. The factors age, birth weight and prematurity were all observed.RESULTS: We evaluated 388 measurements: 159 in the summer and 229 in the winter. Newborn and infants aged from 2 days to 3 months were included. The time of collection of the exam was significant in the measurement of TSH and the mean levels of TSH collected in the summer were somewhat higher than those collected in the winter (p = 0,0001), when analyzed by the t-test, even so when the age was taken into account. Using F-test (p=0,0234) we observed differences between TSH levels in age groups. TSH levels in the newborn with 2 to 7 days of age differ significantly from those of higher age. Birth weight and prematurity do not show interference with the TSH measurement.CONCLUSIONS: Newborn and infant TSH levels were significantly higher in the summer than in the winter.

Verapamil no tratamento da emergencia hipertensiva. / Verapamil in the treatment of hypertensive emergency

Quarenta e cinco pacientes com emergencia hipertensiva foram tratados com verapamil, na dose de 5,48 a 6,68 micrograma/min, administrado por EV em gotejamento continuo de uma solucao contendo 50 mg da droga diluidos em 200 ml de soro glicosado a 5%. Nao se observou alteracao substancial da frequencia cardiaca apos a administracao do farmaco (reducao de 9,7%). Seus efeitos na pressao arterial foram observados ja a partir de 2 min de infusao, com reducao da PA media de 169 mais ou menos 22 para 120 mais ou menos 16 mm/Hg (30%) em 2 hs. Nao houve diferenca entre a reducao da pressao arterial sistolica e diastolica. O controle imediato da pressao arterial, a obtencao de niveis tensionais adequados e a quase ausencia de efeitos colaterais justificam o uso do verapamil para o tratamento das emergencias hipertensivas