Clostridia and necrotizing enterocolitis in preterm neonates.

Necrotizing enterocolitis (NEC) is the most severe life threatening gastrointestinal disease among preterm neonates. NEC continues to account for substantial morbidity and mortality in neonatal intensive care units worldwide. Although its pathogenesis remains incompletely elucidated, NEC is recognized as a multifactorial disease involving intestinal unbalanced inflammatory response, feeding strategies, and bacterial colonization. Epidemiological studies, clinical signs, and animal models support the participation of anaerobic bacteria, particularly clostridia species, in NEC development. Colonization by clostridia seems particularly deleterious. The present review is the opportunity to propose an update on the role of clostridia and NEC.

Valor nutritivo da Coastcross-1 inoculada com Azospirillum brasilense / Nutritive value of Coastcross-1 inoculated with Azospirillum brasilense

Objetivou-se com este trabalho avaliar a taxa de produção de forragem e o valor nutritivo da Coastcross-1, inoculada com Azospirillum brasilense, fertilizada com diferentes doses de N e submetida ao regime de corte. O delineamento experimental foi o de blocos ao acaso, com três repetições, em esquema fatorial. Os fatores foram o uso da inoculação (não inoculada, inoculada somente no plantio e reinoculada no segundo ano), níveis de N (0, 100 e 200kg/ha/ano de N) e os períodos do ano. Avaliaram-se a taxa de produção e a composição botânica da forragem, a composição morfológica, o teor de proteína bruta, a digestibilidade in situ da matéria orgânica e os nutrientes digestíveis totais da Coastcross-1. A inoculação, quando não associada ao adubo nitrogenado, ocasionou aumento na taxa de produção de forragem, na participação da Coastcross-1 e na produção de nutrientes digestíveis totais. Houve redução nos teores de proteína bruta da Coastcross-1, quando a gramínea adubada com 200kg/ha/ano de N foi inoculada. A inoculação tem efeito positivo na Coastcross-1 não submetida à adubação nitrogenada.(AU)

The objective of this study was to evaluate the forage yield rate and nutritive value of Coastcross-1 inoculated with Azospirillum brasilense and fertilized with different levels of nitrogen subjected to cut. The experimental design was randomized blocks with three replications, in a factorial scheme. The factors were the inoculation (without inoculation, inoculated just in the implantation and reinoculated in the second year), levels of nitrogen (0, 100 and 200kg/ha/year of N) and periods of the year. Forage yield rate and botanical composition; morphological composition, crude protein content, in situ digestibility of organic matter and the total digestible nutrients of Coastcross-1 were evaluated. Inoculation increased the forage yield rate, Coastcross-1 contribution and production of total digestible nutrients, when the pasture was not fertilized with N. There was reduction in Coastcross-1 crude protein content when fertilized with 200kg/ha/yr of N. The inoculation has a positive effect on Coastcross-1 not submitted to nitrogen fertilization.(AU)

Produtividade de genótipos de trigo duplo propósito submetidos ao pastejo com vacas em lactação / Productivity of dual-purpose wheat genotypes under grazing with lactating cows

Esta pesquisa foi conduzida com o objetivo de avaliar a produtividade de dois genótipos de trigo de duplo propósito, BRS Tarumã e BRS Umbu, submetidos ao pastejo com vacas em lactação. O delineamento experimental foi o inteiramente ao acaso, com dois tratamentos (genótipos), três repetições (piquetes) e medidas repetidas no tempo (pastejos). Avaliaram-se a precocidade, a composição estrutural dos trigos, as produções de forragem e de biomassa de lâminas foliares, as taxas de acúmulo diário de forragem e de lâminas foliares, a taxa de lotação, as ofertas de forragem e de lâminas foliares, a eficiência de pastejo, o consumo aparente e a produção de grãos. O trigo mais precoce para produção de forragem foi o BRS Umbu. Houve diferença para a produção de forragem (3196 vs. 4143kg MS/ha) e de lâminas foliares (2281 vs. 3205kg MS/ha) para os genótipos BRS Umbu e BRS Tarumã, respectivamente. Valores similares foram encontrados para taxa de lotação (2,26UA/ha); eficiência de pastejo (52,26%), consumo aparente (2,91%) e produção de grãos (1716kg/ha). O genótipo BRS Tarumã é o mais indicado para o manejo de duplo propósito em condições de pastejo com vacas em lactação.(AU)

The objective of this research was to evaluate the productivity of two dual-purpose wheat genotypes BRS Tarumã and BRS Umbu under grazing with lactating cows. The experimental design was completely randomized, with two treatments, three replications (paddocks) and repeated measures (grazing cycles). Studied variables were early growth, the structural composition of wheat, forage production, leaf blade biomass, the stocking rate, the herbage and leaf blade allowance, the grazing efficiency, the herbage intake and grain yield. The earliest genotype for forage production was the BRS Umbu. Differences in herbage yield between BRS Umbu and BRS Tarumã genotypes (3196 vs. 4143kg DM/ha) and leaf blade production (2281 vs. 3205kg DM/ha), respectively, were detected. Similar values between cultivars were found in stocking rate (2,26AU/ha); grazing efficiency (52,26%), herbage intake (2,91 %) and grain yield (1716kg/ha). The BRS Tarumã genotype is the most suitable for dual-purpose under grazing with dairy cows.(AU)

Comparative phenotypic analysis of "Clostridium neonatale" and Clostridium butyricum isolates from neonates.

"Clostridium neonatale" was recently described as a new species within the Cluster I of the Clostridium genus sensu stricto. In this study, we characterized "C. neonatale" isolates (n = 42) and compared their phenotypic properties with those of Clostridium butyricum (n = 26), a close related species. Strains isolated from fecal samples of healthy neonates were tested for different phenotypic characteristics. Compared to C. butyricum, "C. neonatale" showed a significant higher surface hydrophobicity (p = 0.0047), exopolysaccharide production (p = 0.0069), aero-tolerance (p = 0.0222) and viability at 30 °C (p = 0.0006). A lower swimming ability (p = 0.0146) and tolerance against bile (0.3%) (p = 0.0494), acid (pH 4.5) (p < 0.0001), osmolarity (NaCl 5%, p = 0.0188) and temperature at 50 °C (p = 0.0013) characterized "C. neonatale" strains. Our results showed that "C. neonatale" behaves very differently from C. butyricum and suggests specific responses to environmental changes. Besides it is the first study on clinical isolates for these two anaerobic members of the newborns' gut microbiota and broadens our knowledge about their phenotypic traits.

Microbiote intestinal du nouveau-né : impact des antibiotiques.

Antibiotics are involved act directly or indirectly on the intestinal microbiota. The consequences of changing microbiota equilibrium are an increased risk of developing certain pathologies and the emergence of resistant bacteria. In view of the new information accumulated in recent years, a more reasonable use of antibiotics in neonatology is all the more justified. In this sense, the forthcoming publication of new recommendations for the care of early neonatal bacterial infections seems to be a good initiative.

Countrywide spread of OXA-48 carbapenemase in Lebanon: surveillance and genetic characterization of carbapenem-non-susceptible Enterobacteriaceae in 10 hospitals over a one-year period.

OBJECTIVES: To detect, characterize, and assess the genetic clonality of carbapenem-non-susceptible Enterobacteriaceae in 10 Lebanese hospitals in 2012. METHODS: Selected Enterobacteriaceae isolates with reduced susceptibility to carbapenems were subject to phenotypic study including antibiotic susceptibility, cloxacillin effect, modified Hodge test, and activity of efflux pump inhibitor. Carbapenemase genes were detected using PCR; clonal relatedness was studied by pulsed field gel electrophoresis. RESULTS: Out of 8717 Enterobacteriaceae isolated in 2012, 102 (1.2%) showed reduced susceptibility to carbapenems. Thirty-one (70%) of the 44 studied clinical isolates harbored blaOXA-48, including 15 Klebsiella pneumoniae, eight Escherichia coli, four Serratia marcescens, three Enterobacter cloacae, and one Morganella morganii. The majority of OXA-48 producers co-secreted an extended-spectrum beta-lactamase, while one had an acquired AmpC of the ACC type. In the non-OXA-48 producers, carbapenem resistance was attributed to the production of acquired AmpC cephalosporinases of MOX or CIT type, outer membrane impermeability, and/or efflux pump overproduction. DNA fingerprints revealed that OXA-48 producers were different, except for clonal relatedness among four K. pneumoniae, two E. coli, two E. cloacae, and three S. marcescens. CONCLUSIONS: Nosocomial carbapenem-non-susceptible Enterobacteriaceae are moderately spread in Lebanon and the predominant mechanism is OXA-48 production.

Longitudinal analysis and genotyping of infant dominant bifidobacterial populations.

Bifidobacterial population dynamics were investigated by the longitudinal analysis of the dominant population isolated from the feces of young infants. After molecular identification and fingerprinting comparison, clone identity of the consecutive strains belonging to the same species for one individual was performed by pulsed-field gel electrophoresis. The results, obtained from 15 individuals sampled four times over a five-week period suggested a turnover of the dominant bifidobacteria in the population not only at the species but also at its species representative levels. This study provides new insights of the in vivo dynamics of commensal bifidobacteria. It highlights the need to take into consideration the fluctuation of bifidobacterial populations that may occur in one individual in order to investigate reliably the impact of dietary components, such as probiotics or prebiotics, on the intestinal ecosystem.

Tetracycline resistance mediated by tet(W), tet(M), and tet(O) genes of Bifidobacterium isolates from humans.

MICs of tetracyclines were determined for 86 human Bifidobacterium isolates and three environmental strains. The tet(O) gene was found to be absent in these isolates. tet(W) and tet(M) were found in 26 and 7%, respectively, of the Bifidobacterium isolates, and one isolate contained both genes. Chromosomal DNA hybridization showed that there was one chromosomal copy of tet(W) and/or tet(M).

Cytokines in the pruritic papular eruption of HIV.

BACKGROUND: The immunopathogenic mechanism of the pruritic papular eruption (PPE) of patients with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) is poorly understood, and the objective of the present study was to determine the concentration of the serum cytokines interleukin-2 (IL-2), IL-4, IL-5, IL-10, IL-12, and gamma-interferon (gamma-IFN) in an attempt to recognize the pattern of CD4+/CD8+ lymphocytes occurring in this dermatosis. MATERIALS AND METHODS: The study was conducted on 11 HIV-positive PPE patients, matched for sex and age with eight HIV-infected patients with no dermatosis and 10 healthy HIV-negative individuals. Cytokines were quantified by enzyme-linked immunoabsorbent assay (ELISA) using monoclonal antibodies (R & D Systems) and the data were analyzed by the Mann-Whitney, Kruskall-Wallis, and Spearman correlation tests. RESULTS: An increased concentration of IL-2 was observed in both the HIV-positive (77.65 pg/mL, P < 0.001) and PPE (20.42 pg/mL, P < 0.05) groups when compared with the HIV-negative group (9.50 pg/mL). The IL-2 concentration was significantly higher (P < 0.05) in the HIV-positive group than in the PPE group. Similarly, the gamma-IFN concentration was higher in the HIV-positive (14.97 pg/mL) and PPE (12.67 pg/mL) groups when compared with the HIV-negative group (8.58 pg/mL). The IL-12 concentration was similar in the PPE and HIV-positive groups (1.82 and 1.68 pg/mL, respectively), but higher than in the HIV-negative group (1.17 pg/mL). The same occurred with IL-5 (17.78, 17.79, and 15.74 pg/mL, respectively). There was no significant difference in IL-4 concentration among the PPE, HIV-positive, and HIV-negative groups (10.95, 7.88, and 10.16 pg/mL, respectively), and the same was observed for IL-10 (22.41, 21.13, and 20.92, respectively). There was a negative correlation between serum gamma-IFN concentration and peripheral CD4+ lymphocyte number (r = - 0.6256) in the PPE group (P < 0.05). CONCLUSIONS: The lower levels of IL-2 and gamma-IFN and the negative correlation between gamma-IFN and peripheral CD4+ lymphocytes may indicate an early phase of immunosuppression in PPE.

Involvement of an active efflux system in the natural resistance of Pseudomonas aeruginosa to aminoglycosides.

A mutant, named 11B, hypersusceptible to aminoglycosides, tetracycline, and erythromycin was isolated after Tn501 insertion mutagenesis of Pseudomonas aeruginosa PAO1. Cloning and sequencing experiments showed that 11B was deficient in an, at that time, unknown active efflux system that contains homologs of MexAB. This locus also contained a putative regulatory gene, mexZ, transcribed divergently from the efflux operon. Introduction of a recombinant plasmid that carries the genes of the efflux system restored the resistance of 11B to parental levels, whereas overexpression of these genes strongly increased the MICs of substrate antibiotics for the PAO1 host. Antibiotic accumulation studies confirmed that this new system is an energy-dependent active efflux system that pumps out aminoglycosides. Furthermore, this system appeared to function with an outer membrane protein, OprM. While the present paper was being written and reviewed, genes with a sequence identical to our pump genes, mexXY of P. aeruginosa, have been reported to increase resistance to erythromycin, fluoroquinolones, and organic cations in Escherichia coli hosts, although efflux of aminoglycosides was not examined (Mine et al., Antimicrob. Agents Chemother. 43:415-417, 1999). Our study thus shows that the MexXY system plays an important role in the intrinsic resistance of P. aeruginosa to aminoglycosides. Although overexpression of MexXY increased the level of resistance to fluoroquinolones, disruption of the mexXY operon in P. aeruginosa had no detectable effect on susceptibility to these agents.

Use of steroids to monitor alterations in the outer membrane of Pseudomonas aeruginosa.

Testosterone (a strongly hydrophobic steroid) and testosterone hemisuccinate (a negatively charged derivative) were used as probes to investigate alterations in the outer membrane of Pseudomonas aeruginosa. Diffusion rates of the steroids across the lipid bilayer were measured by coupling the influx of these compounds to their subsequent oxidation by an intracellular delta1-dehydrogenase enzyme. Wild-type cells of P. aeruginosa (strain PAO1) were found to be 25 times more permeable to testosterone than to testosterone hemisuccinate. The uptake of the latter compound appeared to be partially dependent on the external pH, thus suggesting a preferential diffusion of the uncharged protonated form across the cell envelope. Using various PAO mutants, we showed that the permeation of steroids was not affected by overexpression of active efflux systems but was increased up to 5.5-fold when the outer membrane contained defective lipopolysaccharides or lacked the major porin OprF. Such alterations in the hydrophobic uptake pathway were not, however, associated with an enhanced permeability of the mutants to the small hydrophilic molecule N,N,N',N'-tetramethyl-p-phenylene diamine. Thirty-six agents were also assayed for their ability to damage the cell surface of strain PAO1, using testosterone as a probe. Polymyxins, rBPI23, chlorhexidine, and dibromopropamidine demonstrated the strongest permeabilizing activities on a molar basis in the presence of 1 mM MgCl2. These amphiphilic polycations increased the transmembrane diffusion of testosterone up to 50-fold and sensitized the PAO1 cells to hydrophobic antibiotics. All together, these data indicated that the steroid uptake assay provides a direct and accurate measurement of the hydrophobic uptake pathway in P. aeruginosa.