Staphylococcus epidermidis in feedings and feces of preterm neonates.

Staphylococcus epidermidis has emerged as the leading agent causing neonatal late-onset sepsis in preterm neonates; although the severity of the episodes caused by this species is often underestimated, it might exert relevant short- and long-term detrimental effects on neonatal outcomes. In this context, the objective of this study was to characterize a collection of S. epidermidis strains obtained from meconium and feces of preterm infants, and to assess the potential role of the enteral feeding tubes as potential reservoirs for this microorganism. A total of 26 preterm infants were enrolled in the study. Meconium and fecal samples were collected weekly during their first month of life (n = 92). Feeding samples were collected after their pass through the enteral feeding tubes (n = 84). S. epidermidis was present in the fecal samples of all the infants in, at least, one sampling time at concentrations ranging from 6.5 to 7.8 log10 CFU/g. Initially, 344 isolates were obtained and pulsed-field gel electrophoresis (PFGE) profiling allowed the reduction of the collection to 101 strains. Among them, multilocus sequence typing (MLST) profiling showed the presence of 32 different sequence types (ST). Globally, most of the STs to hospital-adapted high-risk clones and belonged to clonal complexes (CC) associated to the hospital environment, such as CC2. The virulence gene most commonly detected among the strains was altE. High resistance rates to macrolides and aminoglycosides were detected and 64% of the strains harboured the mecA gene, which was codified in SCCmec types. Our results indicates the existence of a complex and genetically diverse S. epidermidis population in the NICU environment. A better knowledge of S. epidermidis strains may help to devise strategies to avoid their conversion from symbiont to pathobiont microorganisms in the NICUs.

Serratia marcescens colonization in preterm neonates during their neonatal intensive care unit stay.

Background: Nosocomial sepsis is the main problem that preterms have to face during their stay at neonatal intensive care units (NICU). Serratia marcescens is an emerging cause of preterm sepsis but its epidemiology is still largely unknown. Consequently, the aims of this study were to know the rate of preterms colonized by S. marcescens during their stay at the NICU and the characteristics and evolution of the S. marcescens population, including the susceptibility to clinically relevant antibiotics. Methods: Twenty-six preterm infants born with a gestational age ≤ 32 weeks and/or weigh ≤1500 g were included in the study. Samples of meconium and feces (n = 92) were collected during their first month of life of the infants, together with feeding samples after their pass through enteral feeding tubes (n = 37). Samples were inoculated on MacConkey agar plates. The isolates identified as S. marcescens were genotyped using RAPD and PFGE; and antibiotics susceptibility was performed in a Vitek 2 system. Results: A total of 179 S. marcescens isolates were obtained from the samples. PFGE profiling and cluster analysis allowed the classification of the isolates into 7 different S. marcescens clones. PFGE patterns 1 and 3 were the dominant strains in the fecal samples colonizing 31 and 35% of the infants, respectively. Those isolates causing bacteremia in two infants clustered in PFGE pattern 3. Conclusion: S. marcescens is a bacterial species closely associated to the NICU environment. It can be frequently isolated from preterm's feces although only some genetic lineages seem to be associated to sepsis. Enteral feeding tubes act as important reservoirs to keep the S. marcescens population in the NICU. Trial registration: The local ethic committee approved this trial with the reference 09/157.

Bacteriological and Immunological Profiling of Meconium and Fecal Samples from Preterm Infants: A Two-Year Follow-Up Study.

An abnormal colonization pattern of the preterm gut may affect immune maturation and exert a long-term influence on the intestinal bacterial composition and host health. However, follow-up studies assessing the evolution of the fecal microbiota of infants that were born preterm are very scarce. In this work, the bacterial compositions of fecal samples, obtained from sixteen 2-year-old infants were evaluated using a phylogenetic microarray; subsequently, the results were compared with those obtained in a previous study from samples of meconium and feces collected from the same infants while they stayed in the neonatal intensive care unit (NICU). In parallel, the concentration of a wide range of cytokines, chemokines, growth factors and immunoglobulins were determined in meconium and fecal samples. Globally, a higher bacterial diversity and a lower interindividual variability were observed in 2-year-olds' feces, when compared to the samples obtained during their first days of life. Hospital-associated fecal bacteria, that were dominant during the NICU stay, seemed to be replaced, two years later, by genera, which are usually predominant in the healthy adult microbiome. The immune profile of the meconium and fecal samples differed, depending on the sampling time, showing different immune maturation statuses of the gut.

Bacterial Diversity of the Gastric Content of Preterm Infants during Their First Month of Life at the Hospital.

Studies focused on the stomach microbiota are relatively scarce, and most of them are focused on the adult population. The aim of this work is to describe the bacterial communities inhabiting the gastric content (GC) of preterm neonates. For that purpose, GC samples were collected weekly from a total of 13 preterm neonates during their first month of life within their hospital stay. Samples were analyzed by using both culture-dependent and -independent techniques. The former allowed the isolation of bacteria belonging mainly to the genera Enterococcus, Staphylococcus, Streptococcus, Serratia, Klebsiella, and Escherichia. The cultured dominant species in the GC samples during all the hospitalization period were Enterococcus faecalis and Staphylococcus epidermidis. Multilocus sequence typing (MLST) analysis revealed the presence of high-risk clonal complexes associated with the hospital environment, which may colonize enteral feeding tubes. Similarly, the 16S rRNA sequencing showed that Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Corynebacterium, and Propionibacterium were the dominant genera present at 75% of the gastric samples. However, the genera Serratia, Klebsiella, and Streptococcus were the most abundant. Own mother's milk (OMM) and donor milk (DM) were collected after their pass through the external feeding tubes to assess their bacterial content. OMM and DM had a similar bacterial pattern to GC. Based on these data, the GC of preterm neonates is dominated by Proteobacteria and Firmicutes and harbors high-risk bacterial clones, which may colonize enteral feeding tubes, and therefore the feeds that pass through them.

A Standardized and Data Quality Assessed Maternal-Child Care Integrated Data Repository for Research and Monitoring of Best Practices: A Pilot Project in Spain.

We present the results of a pilot project of the Spanish Ministry of Health, Social Services and Equality, envisaged to the development of a national integrated data repository of maternal-child care information. Based on health information standards and data quality assessment procedures, the developed repository is aimed to a reliable data reuse for (1) population research and (2) the monitoring of healthcare best practices. Data standardization was provided by means of two main ISO 13606 archetypes (composed of 43 sub-archetypes), the first dedicated to the delivery and birth information and the second about the infant feeding information from delivery up to two years. Data quality was assessed by means of a dedicated procedure on seven dimensions including completeness, consistency, uniqueness, multi-source variability, temporal variability, correctness and predictive value. A set of 127 best practice indicators was defined according to international recommendations and mapped to the archetypes, allowing their calculus using XQuery programs. As a result, a standardized and data quality assessed integrated data respository was generated, including 7857 records from two Spanish hospitals: Hospital Virgen del Castillo, Yecla, and Hospital 12 de Octubre, Madrid. This pilot project establishes the basis for a reliable maternal-child care data reuse and standardized monitoring of best practices based on the developed information and data quality standards.

Early Gut Colonization of Preterm Infants: Effect of Enteral Feeding Tubes.

OBJECTIVE: The aim of the study was to evaluate the potential colonization of nosocomial bacteria in enteral feeding systems and its effect on early gut colonization of preterm neonates. METHODS: Mother's own milk, donor milk, and preterm formula samples obtained after passing through the external part of the enteral feeding tubes were cultured. In addition, meconium and fecal samples from 26 preterm infants collected at different time points until discharge were cultured. Random amplification polymorphism DNA and pulse field gel electrophoresis were performed to confirm the presence of specific bacterial strains in milk and infant fecal samples. RESULTS: Approximately 4000 bacterial isolates were identified at the species level. The dominant species in both feces from preterm infants and milk samples were Staphylococcus epidermidis, S aureus, Enterococcus faecalis, E faecium, Serratia marcescens, Klebsiella pneumoniae, and Escherichia coli. All of them were present at high concentrations independently of the feeding mode. Random amplification polymorphism DNA and pulse field gel electrophoresis techniques showed that several bacteria strains were found in both type of samples. Furthermore, scanning electron microscopy revealed the presence of a dense bacterial biofilm in several parts of the feeding tubes and the tube connectors. CONCLUSIONS: There is a sharing of bacterial strains between the neonates' gastrointestinal microbiota and the feeding tubes used to feed them.

Bacterial diversity in meconium of preterm neonates and evolution of their fecal microbiota during the first month of life.

The establishment and succession of bacterial communities in infants may have a profound impact in their health, but information about the composition of meconium microbiota and its evolution in hospitalized preterm infants is scarce. In this context, the objective of this work was to characterize the microbiota of meconium and fecal samples obtained during the first 3 weeks of life from 14 donors using culture and molecular techniques, including DGGE and the Human Intestinal Tract Chip (HITChip) analysis of 16S rRNA amplicons. Culture techniques offer a quantification of cultivable bacteria and allow further study of the isolate, while molecular techniques provide deeper information on bacterial diversity. Culture and HITChip results were very similar but the former showed lower sensitivity. Inter-individual differences were detected in the microbiota profiles although the meconium microbiota was peculiar and distinct from that of fecal samples. Bacilli and other Firmicutes were the main bacteria groups detected in meconium while Proteobacteria dominated in the fecal samples. Culture technique showed that Staphylococcus predominated in meconium and that Enterococcus, together with Gram-negative bacteria such as Escherichia coli, Escherichia fergusonii, Klebsiella pneumoniae and Serratia marcescens, was more abundant in fecal samples. In addition, HITChip results showed the prevalence of bacteria related to Lactobacillus plantarum and Streptococcus mitis in meconium samples whereas those related to Enterococcus, Escherichia coli, Klebsiella pneumoniae and Yersinia predominated in the 3(rd) week feces. This study highlights that spontaneously-released meconium of preterm neonates contains a specific microbiota that differs from that of feces obtained after the first week of life. Our findings indicate that the presence of Serratia was strongly associated with a higher degree of immaturity and other hospital-related parameters, including antibiotherapy and mechanical ventilation.

Heating-induced bacteriological and biochemical modifications in human donor milk after holder pasteurisation.

OBJECTIVES: The objectives of the present study were to enumerate and characterize the pathogenic potential of the Bacillus population that may survive holder pasteurisation of human milk and to evaluate the nutritional damage of this treatment using the furosine and lactulose indexes. MATERIALS AND METHODS: Milk samples from 21 donors were heated at 62.5°C for 30 minutes. Bacterial counts, lactose, glucose, myoinositol, lactulose, and furosine were determined before and after the heat treatment. Some B cereus isolates that survived after pasteurisation were evaluated for toxigenic potential. RESULTS: Nonpasteurised milk samples showed bacterial growth in most of the agar media tested. Bacterial survival after pasteurisation was observed in only 3 samples and, in these cases, the microorganisms isolated belonged to the species B cereus. Furosine could not be detected in any of the samples, whereas changes in lactose, glucose, and myoinositol concentrations after holder pasteurisation were not relevant. Lactulose was below the detection limit of the analytical method in nonpasteurised samples, whereas it was found at low levels in 62% of the samples after holder pasteurisation. The lactation period influenced myoinositol content because its concentration was significantly higher in transition milk than in mature or late lactation milk samples. CONCLUSIONS: Holder pasteurisation led to the destruction of bacteria present initially in donor milk samples, except for some B cereus that did not display a high virulence potential and did not modify significantly the concentration of the compounds analyzed in the present study.