Blood metabolites and fecal starch as indicators of feed efficiency of beef cattle in the feedlot / [Metabolitos sanguíneos e amido fecal como indicadores da eficiência alimentar do gado de corte em confinamento]

The use of blood metabolites (BM), fecal starch (FS), and apparent digestion of starch, (ATTSD) as indicators of feed efficiency (FE) in beef cattle in the feedlot was studied. Fourteen bulls were used, originating in an industrial cross, without a defined racial group, with mean body weight of 284.86kg, individually fed, being evaluated in a 42-day confinement system. After the evaluation, the animals were divided into two groups according to the individual FE: high feed efficiency (HE) and low feed efficiency (LE). There was a difference between the groups in the variables FE, feed conversion (FC), final weight (FW), and daily weight gain (DWG). The FE had a positive correlation with DWG, FC, and FW. There was no difference between the groups for the variables BM, FS, and ATTSD, nor was there any correlation between these variables and FE. Considering the feed cost, the HE animals proved more profitable. BM, FS, and ATTSD did not statistically show potential to be used as indicators of FE, despite the evidence of numerical differences of these variables between the different groups, tendency of correlations with FE, and discriminating function with potential assertiveness.(AU)

Foi estudada a utilização dos metabólitos sanguíneos (BM), do amido fecal (FS) e da digestão aparente do amido (ATTSD) como indicadores de eficiência alimentar (FE) em bovinos de corte em confinamento. Utilizaram-se 14 touros, originários de cruzamento industrial, sem grupo racial definido, peso corporal médio de 284,86kg, alimentados individualmente, sendo avaliados em sistema de confinamento por 42 dias. Após a avaliação, dividiram-se os animais em dois grupos, de acordo com a FE individual: alta eficiência alimentar (HE) e baixa eficiência alimentar (LE). Houve diferença entre os grupos nas variáveis FE, conversão alimentar (FC), peso final (FW) e ganho de peso diário (DWG). A FE teve correlação positiva com DWG, FC e FW. Não houve diferença entre os grupos para as variáveis BM, FS e ATTSD, tampouco houve correlação entre essas variáveis e a FE. Considerando-se o custo alimentar, os animais HE mostraram-se mais lucrativos. BM, FS e ATTSD não mostraram, estatisticamente, potencial para serem utilizados como indicadores de FE, apesar da evidência de diferenças numéricas dessas variáveis entre os diferentes grupos, tendência de correlações com a FE e de função discriminante com potencial assertividade.(AU)

A combined enrichment/polymerase chain reaction based method for the routine screening of Streptococcus agalactiae in pregnant women

Group B Streptococcus (GBS) is the most common cause of life-threatening infection in neonates. Guidelines from CDC recommend universal screening of pregnant women for rectovaginal GBS colonization. The objective of this study was to compare the performance of a combined enrichment/PCR based method targeting the atr gene in relation to culture using enrichment with selective broth medium (standard method) to identify the presence of GBS in pregnant women. Rectovaginal GBS samples from women at ¡Ý36 weeks of pregnancy were obtained with a swab and analyzed by the two methods. A total of 89 samples were evaluated. The prevalence of positive results for GBS detection was considerable higher when assessed by the combined enrichment/PCR method than with the standard method (35.9% versus 22.5%, respectively). The results demonstrated that the use of selective enrichment broth followed by PCR targeting the atr gene is a highly sensitive, specific and accurate test for GBS screening in pregnant women, allowing the detection of the bacteria even in lightly colonized patients. This PCR methodology may provide a useful diagnostic tool for GBS detection and contributes for a more accurate and effective intrapartum antibiotic and lower newborn mortality and morbidity.

When the resistance gets clingy: Pseudomonas aeruginosa harboring metallo-ß-lactamase gene shows high ability to produce biofilm.

The ability to produce biofilm and the presence of metallo-ß-lactamase (MBL) among Pseudomonas aeruginosa isolates were evaluated. A total of 91 isolates were recovered from sputa of patients with (CF, n = 44) and without (non-CF, n = 47) cystic fibrosis diagnosis. Seventy-nine (86.8%; 95% CI 78.3-92.3%) were biofilm producers. Interestingly, all isolates harboring MBL showed ability (most strong or moderate) to produce biofilm in vitro. We alert to an "overlapping of mechanisms" that together represent an even greater challenge for the treatment of pulmonary infections by P. aeruginosa.

Evaluation of biofilm production by Pseudomonas Aeruginosa isolates recovered from cystic fibrosis and non-cystic fibrosis patients

Cystic fibrosis (CF) patients typically suffer of persistent and recurrent lung infections caused by Pseudomonas aeruginosa that many times possess ability for the biofilm production. Here, biofilm production among P. aeruginosa isolates recovered from sputum of CF and non-CF patients was evaluated. Most isolates were biofilm-producing independently of the patient's condition.

Evaluation of biofilm production by Pseudomonas Aeruginosa isolates recovered from cystic fibrosis and non-cystic fibrosis patients.

Cystic fibrosis (CF) patients typically suffer of persistent and recurrent lung infections caused by Pseudomonas aeruginosa that many times possess ability for the biofilm production. Here, biofilm production among P. aeruginosa isolates recovered from sputum of CF and non-CF patients was evaluated. Most isolates were biofilm-producing independently of the patient's condition.

Enterococcus gallinarum carrying the vanA gene cluster: first report in Brazil

In 2000, Enterococcus faecalis resistant to vancomycin was first reported at a tertiary hospital in Porto Alegre, southern Brazil. The resistance spread to other hospitals and surveillance programs were established by hospital infection committees to prevent the spread of vancomycin-resistant enterococci. In February 2002, an isolate initially identified at the genus level as Enterococcus was obtained by surveillance culture (rectal swab) from a patient admitted to a hospital for treatment of septic arthritis in the shoulder. The isolate proved to be resistant to vancomycin by the disc diffusion method and confirmed by an E-test resulting in a minimal inhibitory concentration of > ou = 256 µg/ml. This isolate was sent to a reference laboratory (Laboratório Especial de Bacteriologia e Epidemiologia Molecular, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP) for further study and proved to be an E. gallinarum by the polymerase chain reaction (PCR) using specific primers for the species. Due to the phenotype of unusually high vancomycin resistance, the isolate presumably had the resistance genes (vanA and vanB) and this was confirmed by PCR, which indicated the presence of the vanA gene. A 10.8-kb Tn1546-related transposon was also identified by long-PCR. Interspecies transfer of the vancomycin-resistance gene from the donor E. gallinarum was performed in a successful conjugation experiment in vitro, using E. faecium GE-1 and E. faecalis JH22 as receptors. This is the first report of the detection of a vanA determinant naturally acquired by E. gallinarum in Brazil, indicating the importance of characterizing VRE by both phenotype and genotype methods.

Typing of Pseudomonas aeruginosa from hospitalized patients: a comparison of susceptibility and biochemical profiles with genotype

Typing techniques are essential for understanding hospital epidemiology, permitting the elucidation of the source of infection and routes of bacterial transmission. Although DNA-based techniques are the gold standard for the epidemiological study of Pseudomonas aeruginosa, antibiotic profiles and biochemical results are used because they are easy to perform and to interpret and relatively inexpensive. Antibiotypes (susceptibility profiles) and biotypes (biochemical profiles) were compared to genotypes established by DNA restriction enzyme analysis in 81 clinical isolates of P. aeruginosa from three hospitals in Porto Alegre, Brazil. The epidemiological relationship among patients was also evaluated. Susceptibility and restriction profiles were discrepant in more than 50 percent of the cases, and many antibiotypes were observed among isolates from the same genotype. Furthermore, susceptibility profiles did not allow the distinction of isolates from unrelated genotypes. Since a large number of isolates (63 percent) yielded the same biochemical results, only 10 biotypes were detected, showing that this typing method has a low discriminatory power. On the other hand, DNA restriction enzyme typing allowed us to establish 71 distinct types. Epidemiological data about the relation among P. aeruginosa isolates were not conclusive. The results of the present study indicate that the only method that can establish a clonal relation is DNA restriction enzyme typing, whereas the other methods may cause misleading interpretations and are inadequate to guide proper infection control measures.

Heterogeneity of Pseudomonas aeruginosa in Brazilian cystic fibrosis patients.

The aim of this study was to assess the diversity and genomic variability of Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients being treated at a university hospital in Brazil. Ninety-seven isolates of P. aeruginosa from 43 CF patients were characterized by macrorestriction analysis of chromosomal DNA by pulsed-field gel electrophoresis (PFGE) and tested for susceptibility to 20 antimicrobial agents by broth microdilution. It was possible to evaluate single isolates from 20 patients and multiple isolates (two to seven) from 23 patients collected during a 22-month period. Among all of the unrelated patients, we detected only one pair of patients sharing a common strain. Among the 77 isolates from 23 patients who had multiple isolates analyzed, we identified 37 major types by PFGE, and five different colonization patterns were recognized. The isolates were susceptible to several antimicrobial agents, although consecutive isolates from the same patient may display differences in their susceptibilities. Mucoid isolates were more resistant (P < 0.001) than nonmucoid isolates to five antibiotics. Our results indicate that CF patients remain colonized by more than one strain of P. aeruginosa for long periods of time. In addition, the finding of several different genotypes in the same patient suggests that the colonizing strain may occasionally be replaced.

Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae

In order to evaluate the resolving power of several typing methods to identify relatedness among Brazilian strains of Enterobacter cloacae, we selected twenty isolates from different patients on three wards of a University Hospital (Orthopedics, Nephrology, and Hematology). Traditional phenotyping methods applied to isolates included biotyping, antibiotic sensitivity, phage-typing, and O-serotyping. Plasmid profile analysis, ribotyping, and macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) were used as genotyping methods. Sero- and phage-typing were not useful since the majority of isolates could not be subtyped by these methods. Biotyping, antibiogram and plasmid profile permitted us to classify the samples into different groups depending on the method used, and consequently were not reliable. Ribotyping and PFGE were significantly correlated with the clinical epidemiological analysis. PFGE did not type strains containing nonspecific DNase. Ribotyping was the most discriminative method for typing Brazilian isolates of E. cloacae

Complementation of methionine auxotrophs of Pseudomonas aeruginosa from cystic fibrosis.

Auxotrophic Pseudomonas aeruginosa are exclusive to respiratory infections in cystic fibrosis (CF) and bronchiectatic patients, and isolates require specific amino acids for growth on minimal media, particularly methionine. Since auxotrophic and prototrophic P. aeruginosa from CF are identical by genotyping, we investigated the genetic events leading to methionine auxotrophy (Met-). Most (10/13) Met- strains had the same pattern of growth on methionine precursors and required methionine exclusively for growth. Back mutation to prototrophy was very low (frequencies 10(-8) to <10(-10)). Complementation of the mutations leading to auxotrophy was achieved for five strains with a genomic library of P. aeruginosa PAO1. Strains with different patterns of growth on methionine precursors were complemented by clones with different restriction patterns, while identical clones complemented strains with the same pattern of growth on methionine precursors. Methionine auxotrophy in P. aeruginosa from CF results from stable chromosomal mutations, and the commonest defect is probably in gene(s) encoding enzymes that convert homocysteine to methionine.

The high amino-acid content of sputum from cystic fibrosis patients promotes growth of auxotrophic Pseudomonas aeruginosa.

Many isolates of Pseudomonas aeruginosa from cystic fibrosis (CF) patients are auxotrophic and require amino acids for growth. A quantitative assay was used to determine the total content of free amino acids of sputum sol-phase extracts from CF and non-CF patients to assess the presence of amino acids in the airway. CF patients colonised with auxotrophic P. aeruginosa had a higher sputum amino-acid content (mean 6.77 mg/ml) than those colonised with prototrophs (mean 3.77 mg/ml); overall, CF specimens (mean 5.70 mg/ml) had a higher amino-acid content than non-CF samples (2.52 mg/ml). The amino-acid profile of sputum extracts was assessed by one-dimensional thin layer chromatography (TLC). Several amino acids were identified in the extracts, in particular, leucine, isoleucine, phenylalanine, tyrosine, alanine, serine and methionine or valine or both. All sputum specimens except two (which contained < 1.5 mg of amino acids/ml), promoted the growth, of 34 auxotrophic strains of P. aeruginosa from CF patients in a minimal medium. These results indicate, therefore, that amino acids are plentiful in the sputum of CF patients and are able to supply the requirements of auxotrophic strains. It is suggested that the increased amino-acid content in the airways of CF patients plays a significant role in the selection and maintenance of nutritionally deficient P. aeruginosa.

Auxotrophy of Burkholderia (Pseudomonas) cepacia from cystic fibrosis patients.

The nutritional status of 89 isolates of Burkholderia cepacia from 81 cystic fibrosis (CF) patients was evaluated. Forty of the isolates, from 38 patients, were not able to grow in a minimal medium containing glucose and mineral salts only and were thus auxotrophs. In contrast, all of 29 isolates from non-CF (clinical and environmental) sources were prototrophic. Addition of a pool of amino acids to the minimal medium was sufficient to promote growth of all tested CF auxotrophic isolates. Indeed, phenylalanine, tyrosine, cysteine, methionine, and histidine alone or in combination were required for growth by the majority of the nutritionally deficient B. cepacia isolates. Furthermore, extracts of sputum from CF patients, when added to minimal medium, promoted growth of 29 auxotrophic B. cepacia isolates regardless of their amino acid requirements. Finally, auxotrophic and prototrophic isolates from the same patient exhibited a conserved genotype, as determined by macrorestriction analysis of chromosomal DNA. These results suggest that the auxotrophic mutants are selected from the prototrophic population and maintained by the nutritionally rich environment of the CF airways.

Auxotrophic variants of Pseudomonas aeruginosa are selected from prototrophic wild-type strains in respiratory infections in patients with cystic fibrosis.

Twenty-four nutritionally dependent (auxotrophic) Pseudomonas aeruginosa strains were isolated from 20 cystic fibrosis (CF) patients and tested for their amino acid requirements. Two different methods were necessary to identify the nutritional status of all isolates. Methionine was the most common single amino acid required (9 of 24 isolates), followed by leucine and arginine or ornithine. In total, a requirement for 12 different compounds or combination of compounds was demonstrated. Auxotrophic and prototrophic pairs of isolates from the same patient were compared by macrorestriction analysis of DNA in pulsed-field gel electrophoresis. Thirteen of 18 pairs analyzed presented identical restriction fragment length polymorphism profiles following digestion of DNA with XbaI. Three of the remaining pairs showed percentage similarities of 77, 91, and 98%, and the profiles of two pairs could not be compared because of the excessive degradation of their DNA. These results suggest that auxotrophic and prototrophic P. aeruginosa isolates colonizing the same CF patient constitute an isogenic group and raise the possibility that auxotrophs are selected from the prototrophic population during the course of pulmonary infection in CF patients.

Defective repair of radiation-induced chromosomal damage in scid/scid mice.

The murine severe combined immunodeficiency (scid) mutation interferes with normal recombination of immunoglobulin and T-cell receptor genes. This immunologic defect results in a lack of fully differentiated B and T cells in scid/scid mice. Animals homozygous for the scid mutation also display increased sensitivity to the damaging effects of ionizing radiation. We report here our observations of high frequencies of radiation-induced chromatid interchanges and intrachanges in bone marrow cells and fibroblasts from scid/scid mice. The presence of these aberrant chromosome structures suggests that a delay in strand rejoining underlies the increased sensitivity of scid/scid mice to ionizing radiation. The scid mutation may provide important clues for understanding the relationship between mitotic recombination and DNA repair in higher eukaryotic cells.