Intravenous antibiotics in preterm infants have a negative effect upon microbiome development throughout preterm life.

BACKGROUND: Intestinal dysbiosis is implicated in the origins of necrotising enterocolitis and late-onset sepsis in preterm babies. However, the effect of modulators of bacterial growth (e.g. antibiotics) upon the developing microbiome is not well-characterised. In this prospectively-recruited, retrospectively-classified, case-control study, high-throughput 16S rRNA gene sequencing was combined with contemporaneous clinical data collection, to assess the within-subject relationship between antibiotic administration and microbiome development, in comparison to preterm infants with minimal antibiotic exposure. RESULTS: During courses of antibiotics, diversity progression fell in comparison to that seen outside periods of antibiotic use (-0.71units/week vs. + 0.63units/week, p < 0.01); Enterobacteriaceae relative abundance progression conversely rose (+ 10.6%/week vs. -8.9%/week, p < 0.01). After antibiotic cessation, diversity progression remained suppressed (+ 0.2units/week, p = 0.02). CONCLUSIONS: Antibiotic use has an acute and longer-lasting impact on the developing preterm intestinal microbiome. This has clinical implications with regard to the contribution of antibiotic use to evolving dysbiosis, and affects the interpretation of existing microbiome studies where this effect modulator is rarely accounted for.

Perinatal inflammation influences but does not arrest rapid immune development in preterm babies.

Infection and infection-related complications are important causes of death and morbidity following preterm birth. Despite this risk, there is limited understanding of the development of the immune system in those born prematurely, and of how this development is influenced by perinatal factors. Here we prospectively and longitudinally follow a cohort of babies born before 32 weeks of gestation. We demonstrate that preterm babies, including those born extremely prematurely (<28 weeks), are capable of rapidly acquiring some adult levels of immune functionality, in which immune maturation occurs independently of the developing heterogeneous microbiome. By contrast, we observe a reduced percentage of CXCL8-producing T cells, but comparable levels of TNF-producing T cells, from babies exposed to in utero or postnatal infection, which precedes an unstable post-natal clinical course. These data show that rapid immune development is possible in preterm babies, but distinct identifiable differences in functionality may predict subsequent infection mediated outcomes.

Effects of oral probiotic supplements on vaginal microbiota during pregnancy: a randomised, double-blind, placebo-controlled trial with microbiome analysis.

OBJECTIVE: To determine the effects on the vaginal microbiota of an oral probiotic preparation administered from early pregnancy. DESIGN: Randomised, double blind, placebo-controlled trial. SETTING: Four maternity units in the UK. POPULATION: Women aged 16 years or older recruited at 9-14 weeks' gestation. METHODS: Participants were randomly allocated to receive oral capsules of probiotic containing Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 each at 2.5 × 109 colony-forming units (CFUs) or placebo once daily from recruitment until the end of pregnancy. MAIN OUTCOME MEASURE: Rates of bacterial vaginosis (BV, defined as Nugent score ≥7) at 18-20 weeks' gestation compared by logistic regression adjusted for possible confounders. RESULTS: The primary analysis included 78% (238/304) of participants who initially consented (probiotic group 123, placebo group 115). Of these participants, 95% (227/238) reported an intake of 93% or more of the required number of capsules. The rates of BV did not differ between groups at 18-20 weeks' gestation (15% (19/123) in the probiotic group vs. 9% (10/115) in the placebo group, adjusted odds ratio 1.82, 95% confidence interval 0.64-5.19). There were also no differences between the groups in the proportion of women colonised with the probiotic strains, Escherichia coli, group B streptococci or other vaginal microbiota. There were no differences in the alpha diversity or composition of the bacterial communities between or within the probiotic and placebo groups at 9-14 and 18-20 weeks' gestation. CONCLUSIONS: Oral probiotics taken from early pregnancy did not modify the vaginal microbiota. TWEETABLE ABSTRACT: The oral probiotic preparation used in this study does not prevent BV in pregnant women.

Stimulation of growth hormone by kisspeptin antagonists in ewes.

Kisspeptin signalling is indispensable for fertility, stimulating gonadotropin-releasing hormone (GnRH) secretion and mediating gonadal steroid feedback on GnRH neurons. Moreover, kisspeptin neurons have been implicated in other non-reproductive neuroendocrine roles. Kisspeptin appears to also regulate growth hormone secretion but much of the data appear contradictory. We sought to clarify a potential role of kisspeptin in growth hormone (GH) regulation by examining the effect of kisspeptin antagonists on GH secretion in ewes under various physiological conditions. Our data show clear and robust increases in GH secretion following lateral ventricle or third ventricle infusion of kisspeptin antagonists p-234 and p-271 in either ovariectomized or anestrous ewes. Central infusion of kisspeptin-10 had no effect on GH secretion. To determine the level at which kisspeptin may influence GH secretion, we examined expression of the cognate kisspeptin receptor, GPR54, in pituitary cells and showed by immunocytochemistry that the majority of somatotropes express GPR54 while expression was largely negative in other pituitary cells. Overall, we have demonstrated that blocking kisspeptin signalling by antagonists stimulates GH secretion in ewes and that this is likely mediated by inhibiting endogenous kisspeptin activation of GPR54 expressed on somatotropes. The findings suggest that endogenous kisspeptin inhibits GH secretion through GPR54 expressed on somatotropes.

Antibiotic treatment duration and prevention of complications in neonatal Staphylococcus aureus bacteraemia.

BACKGROUND: In adults with Staphylococcus aureus bacteraemia, short duration of effective antibiotic treatment is associated with increased risk of complications and recurrence. The optimum duration of treatment for neonates is unknown and practice varies widely. AIM: To relate the duration of treatment of neonatal S. aureus bacteraemia to prevention of complications and recurrence. METHODS: Retrospective cohort study of confirmed S. aureus bacteraemia occurring over a 10 year period in two large tertiary neonatal units. Neonatal patients developing confirmed S. aureus bacteraemia between birth and discharge from the neonatal unit were identified from microbiology department records. Clinical details obtained from case notes included demographics, duration of antibiotics and clinical outcomes. Recurrence was determined from laboratory and clinical records. Adverse outcomes were related to duration of antibiotic therapy. FINDINGS: A total of 90 infants had S. aureus bacteraemia, of which six were meticillin-resistant S. aureus (7%). Median gestation was 27 weeks (range: 23-41), birth weight 846 g (434-3840) and postnatal age 16 days (0-116). Adverse outcomes were found in 44%, with death in 8%. Median duration of appropriate antibiotics was 19 days (range: 0-54). There were no cases of recurrent bacteraemia after finishing antibiotics. There was no relationship between antibiotic duration and complications. CONCLUSION: Neonatal S. aureus bacteraemia mainly affected preterm neonates and had a significant morbidity and mortality. Recurrent bacteraemia was rare, irrespective of treatment duration. For neonatal unit patients with S. aureus bacteraemia, antibiotic therapy for 14 days in uncomplicated cases may be sufficient to prevent recurrence, with longer treatment justified if there is inadequate source control.

Contrasting patterns of clonality and fine-scale genetic structure in two rare sedges with differing geographic distributions.

For plants with mixed reproductive capabilities, asexual reproduction is more frequent in rare species and is considered a strategy for persistence when sexual recruitment is limited. We investigate whether asexual reproduction contributes to the persistence of two co-occurring, rare sedges that both experience irregular seed set and if their differing geographic distributions have a role in the relative contribution of clonality. Genotypic richness was high (R=0.889±0.02) across the clustered populations of Lepidosperma sp. Mt Caudan and, where detected, clonal patches were small, both in ramet numbers (⩽3 ramets/genet) and physical size (1.3±0.1 m). In contrast, genotypic richness was lower in the isolated L. sp. Parker Range populations, albeit more variable (R=0.437±0.13), with genets as large as 17 ramets and up to 5.8 m in size. Aggregated clonal growth generated significant fine-scale genetic structure in both species but to a greater spatial extent and with additional genet-level structure in L. sp. Parker Range that is likely due to restricted seed dispersal. Despite both species being rare, asexual reproduction clearly has a more important role in the persistence of L. sp. Parker Range than L. sp. Mt Caudan. This is consistent with our prediction that limitations to sexual reproduction, via geographic isolation to effective gene exchange, can lead to greater contributions of asexual reproduction. These results demonstrate the role of population isolation in affecting the balance of alternate reproductive modes and the contextual nature of asexual reproduction in rare species.

Perinatal vertical transmission of antibiotic-resistant bacteria: a systematic review and proposed research strategy.

BACKGROUND: Antibiotic-resistant bacteria contribute to both early- and late-onset sepsis and outbreaks in neonatal intensive care units (NICUs). The extent to which vertical transmission of these resistant bacteria contributes to colonisation or infection of vulnerable infants in NICUs is unclear. Risk factors for vertical transmission of antibiotic-resistant bacteria are not well described. OBJECTIVES: To identify studies describing vertical transmission of antibiotic-resistant bacteria, risk factors for transmission and the impact of colonisation on neonatal outcomes. SEARCH STRATEGY: EMBASE, CINAHL, Cochrane, PubMed, and MEDLINE databases were searched using selected terminology. Titles and abstracts were screened by two reviewers. Selected papers were reviewed in full by two individuals to ascertain whether they fulfilled the inclusion criteria. SELECTION CRITERIA: Any original article investigating perinatal vertical transmission of antibiotic-resistant bacteria between a mother and neonate was included. DATA COLLECTION AND ANALYSIS: Data were extracted on study design, organism, antibiotic resistance, and means of ascertaining vertical transmission. MAIN RESULTS: Five papers out of 4839 titles fulfilled the inclusion criteria. Four studies were predominantly observational and one was a case report. Each demonstrated perinatal transmission. No study reported risk factors for the transmission of resistant bacteria or the impact of colonisation on neonatal outcomes. AUTHOR'S CONCLUSIONS: There is an absence of research into the perinatal transmission of resistant organisms despite the potential implications of such a situation. We outline objectives that need to be addressed in future research and describe a study design to ascertain the prevalence and risk factors for vertical transmission.

Diversity and stability of cultured vaginal lactobacilli in pregnant women from a multi-ethnic urban UK population.

AIM: To determine the diversity and stability of cultured vaginal lactobacilli in a multi-ethnic population of pregnant women. METHODS AND RESULTS: A single-centre, prospective, cohort study was performed in a tertiary perinatal centre in East London, UK. Self-collected vaginal swabs at 13 and 20 weeks gestation were obtained from women attending for routine antenatal care and cultured for lactobacilli. In women who provided both swabs, 37 of 203 (18%) had no lactobacilli cultured at either time. Only 53 (26%) had the same species at both times. Black women were less likely to have lactobacilli cultured at 13 weeks (P = 0·014), and Black and Asian women were less likely to have lactobacilli cultured at 20 weeks (P = 0·002) compared with those in the White and Other groups. CONCLUSIONS: Significant differences exist between ethnic groups in the carriage and stability of vaginal lactobacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: These differences have implications for the design of interventions aimed at normalizing the vaginal microbiota in pregnant women.

Genetic connectivity and diversity in inselberg populations of Acacia woodmaniorum, a rare endemic of the Yilgarn Craton banded iron formations.

Historically rare plant species with disjunct population distributions and small population sizes might be expected to show significant genetic structure and low levels of genetic diversity because of the effects of inbreeding and genetic drift. Across the globe, terrestrial inselbergs are habitat for rich, often rare and endemic flora and are valuable systems for investigating evolutionary processes that shape patterns of genetic structure and levels of genetic diversity at the landscape scale. We assessed genetic structure and levels of genetic diversity across the range of the historically rare inselberg endemic Acacia woodmaniorum. Phylogeographic and genetic structure indicates that connectivity is not sufficient to produce a panmictic population across the limited geographic range of the species. However, historical levels of gene flow are sufficient to maintain a high degree of adaptive connectivity across the landscape. Genetic diversity indicates gene flow is sufficient to largely counteract any negative genetic effects of inbreeding and random genetic drift in even the most disjunct or smallest populations. Phylogeographic and genetic structure, a signal of isolation by distance and a lack of evidence of recent genetic bottlenecks suggest long-term stability of contemporary population distributions and population sizes. There is some evidence that genetic connectivity among disjunct outcrops may be facilitated by the occasional long distance dispersal of Acacia polyads carried by insect pollinators moved by prevailing winds.

Probiotics: a new frontier for infection control.

Probiotics are live micro-organisms administered to provide health benefits. Probiotics are being increasingly used in healthcare contexts both in research studies and routine practice, for example in neonatal intensive care. Currently there is a paucity of guidelines or regulations governing the mitigation of infection risks associated with the use of probiotics in clinical practice. We propose a number of recommendations to mitigate risks. These include the communication of probiotic use to appropriate stakeholders, ensuring that routine laboratories can identify and test the susceptibility of probiotic strains, assuring standards for preparation and administration, and ensuring surveillance designed to capture adverse events.

Rapid identification of bacteria from bioMérieux BacT/ALERT blood culture bottles by MALDI-TOF MS.

Several studies have reported poor results when trying to identify microorganisms directly from the bioMérieux BacT/ALERT blood culture system using matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry. The aim of this study is to evaluate two new methods, Sepsityper and an enrichment method for direct identification of microorganisms from this system. For both methods the samples were processed using the Bruker Microflex LT mass spectrometer (Biotyper) using the Microflex Control software to obtain spectra. The results from direct analysis were compared with those obtained by subculture and subsequent identification. A total of 350 positive blood cultures were processed simultaneously by the two methods. Fifty-three cultures were polymocrobial or failed to grow any organism on subculture, and these results were not included as there was either no subculture result, or for polymicrobial cultures it was known that the Biotyper would not be able to distinguish the constituent organisms correctly. Overall, the results showed that, contrary to previous reports, it is possible to identify bacteria directly from bioMérieux blood culture bottles, as 219/297 (74%) correct identifications were obtained using the Bruker Sepsityper method and 228/297 (77%) were obtained for the enrichment method when there is only one organism was present. Although the enrichment method was simpler, the reagent costs for the Sepsityper method were approximately pound 4.00 per sample compared to pound 0.50. An even simpler and cheaper method, which was less labour-intensive and did not require further reagents, was investigated. Seventy-seven specimens from positive signalled blood cultures were analysed by inoculating prewarmed blood agar plates and analysing any growth after 1-, 2- and 4-h periods of incubation at 37 degrees C, by either direct transfer or alcohol extraction. This method gave the highest number of correct identifications, 66/77 (86%), and was cheaper and less labour-intensive than either of the two above methods.

Pyoderma gangrenosum-like ulcer caused by Helicobacter cinaedi in a patient with x-linked agammaglobulinaemia.

Cutaneous lesions of the legs have been linked to Helicobacter species in a number of patients with X-linked agammaglobulinaemia (XLA), a primary immunodeficiency. We describe a 26-year-old patient with XLA, who was referred with an extensive skin ulcer that enlarged gradually over the course of 7 years. The ulcer resembled pyoderma gangrenosum (PG), and extended from below the knee to the ankle. The man (who has sex with men) was negative for human immunodeficiency virus. Helicobacter cinaedi was identified by 16S ribosomal (r)DNA PCR analysis from a biopsy of the lesion. This fastidious organism has been implicated previously in causing unexplained skin macules in one other patient with XLA. We suggest that early consideration of infection with Helicobacter species in immunocompromised patients who present with unexplained cutaneous lesions is important, as a prolonged antibiotic course can lead to clinical improvement.

Clinical features of psoroptic mange in cattle in England and Wales.

The clinical signs, treatments used and spread of psoroptic mange in cattle from October 2007 until March 2011 are described. The disease was first diagnosed in South West Wales, having not been reported in Great Britain since the 1980s. The likely source was identified as a farm that had imported two animals from mainland Europe in the summer of 2006. Since that time, disease has been diagnosed on a further 22 premises, the majority in South West Wales but also in South East and Mid Wales and on one farm in England. Bought in animals harbouring the Psoroptes species mite but not showing clinical signs were considered the greatest risk of introducing the infestation into a herd. This, together with the difficulties of treatment to eliminate the parasite, means that it is unlikely that this outbreak has been controlled. There is also a continuing threat of importing the disease from abroad. The disease is not notifiable in the UK.

Risk-adjusted comparisons of bloodstream infection rates in neonatal intensive-care units.

Comparisons of bloodstream infection (BSI) rates between neonatal intensive-care units (NICUs) should take into account differences in babies' vulnerability and invasive procedures that can introduce infection. Our aim was to investigate which risk factors recorded in routine records should be adjusted for when NICUs are compared. This was a retrospective cohort study using routine records for two London NICUs. We analysed rates of BSI with Poisson regression models. The level of neonatal care used by the National Health Service was the strongest predictor of BSI incidence. The rate ratios for BSI, adjusted for birthweight, inborn/outborn status, and postnatal age, were 3.15 (95% CI 2.01-4.94) for intensive care and 6.58 (95% CI 4.18-10.36) for high-dependency care, relative to special care. Total parenteral nutrition was significantly associated with BSI incidence, but explained less of the variance among babies than level of care. A case-control study with the same dataset gave similar results. Further multicentre studies are required to confirm our predictive model. Until then, we recommend that comparisons of BSI rates between NICUs should include adjustments for level of care, birthweight, inborn/outborn status, and postnatal age, with the use of routinely recorded standardized measures in hospital administrative data.

Levels of periodontal pathogens in neonatal gastric aspirates and possible maternal sites of origin.

Maternal periodontal infection has been recognized as a risk factor for preterm and low birthweight infants. It is suspected that pathogens causing periodontal disease may translocate to the amniotic cavity and so contribute to triggering an adverse pregnancy outcome. This study aimed to determine levels and proportions of periodontal bacteria in neonatal gastric aspirates obtained from complicated pregnancies and the respective maternal oral and vaginal samples using a quantitative polymerase chain reaction approach, and also to determine the origin of the neonate's bacteria by sequence comparisons between the three sites. Aggregatibacter actinomycetemcomitans and Tannerella forsythia were not observed in the neonates or in the women's vaginas. Interestingly, Porphyromonas gingivalis was identified in the neonates in two samples (2.98E+02 and 1.75E+02 cells ml(-1)) and in association with Fusobacterium nucleatum, which was observed at high prevalence (10%) and at high levels reaching up to 2.32E+03 cells ml(-1). Although F. nucleatum was also present in the vaginal samples, the results demonstrated that the neonatal strains were more likely to originate from the mother's oral cavity than to be vaginal strains.

Patient rights and healthcare-associated infection.

The Universal Declaration of Human Rights was adopted by the United Nations in 1948, and since that time, human rights have become widely recognized and legally enforceable in many countries. Patient rights are now included in healthcare constitutions, such as that of the English National Health Service, and in professional codes of practice. Patient rights have a number of implications for the control of healthcare-associated infections (HCAI), including: (1) justification for infection control over and above economic benefit; (2) focus and emphasis on the individual patient experience; (3) identification of some of the actions taken to control infection as breaches of rights; (4) bridging professional, infection control and public health ethics; (5) a requirement to specify the conditions under which rights can be breached; and (6) grounds for those seeking compensation for HCAI. Assuring patient rights has the potential to improve the patient experience, and in so doing, improve public confidence in healthcare provision and providers.

Subcutaneous anthrax in three intravenous drug users: a new clinical diagnosis.

Anthrax is extremely rare in the western world but is endemic to areas of south and central Asia. In early 2010 an outbreak was identified in heroin-injecting intravenous drug users in the United Kingdom and Europe. Afghanistan is currently the principal source of heroin which reaches the United Kingdom. When anthrax occurs, cutaneous disease accounts for over 95% of cases. At least 47 cases with 13 deaths have been confirmed so far. We present three cases presenting during this time with marked swelling, one resulting in compartment syndrome but all with an absence of the expected cutaneous appearances. We suggest that rather than cutaneous anthrax, these patients represent a new subcutaneous presentation of anthrax.

Accuracy of bacterial DNA testing for central venous catheter-associated bloodstream infection in children with cancer.

BACKGROUND: Central venous catheters (CVCs) are widely used for children with cancer and are a major risk factor for bloodstream infection. Early and specific diagnosis of CVC-associated bloodstream infection allows early targeted treatment, reducing the risk of CVC removal and avoiding the operative risks and trauma of reinsertion, but peripheral vein sampling, as used in adults, improves specificity but is not usually acceptable in children. OBJECTIVE: To improve the detection and treatment of CVC-associated bloodstream infection in children (aged 0-18 years) with cancer admitted with fever. METHODS: There were four main studies: (1) evaluation of the diagnostic accuracy of a quantitative molecular method for the detection of bacterial deoxyribonucleic acid (DNA), based solely on blood samples drawn through the CVC; (2) analysis of the prognostic risk of CVC removal and duration of intravenous (i.v.) antibiotic treatment days in relation to presenting clinical features, blood culture results and bacterial DNA test results; (3) systematic reviews of treatment options for CVC-associated infection and a questionnaire survey of current practice in paediatric oncology centres; (4) evaluation of the clinical effectiveness of different test-treatment strategies to reduce i.v. antibiotic treatment days and unnecessary CVC removals. RESULTS: (1) The bacterial DNA test detected two-thirds [95% confidence interval (CI) 44% to 83%] of children classified with probable CVC-associated infection - specificity was 88% (95% CI 84% to 92%). Although high bacterial DNA concentrations were associated with subsequent CVC removal and long duration of i.v. antibiotic treatment, the test did not improve the prediction of these outcomes over and above clinical signs of CVC-associated infection combined with blood culture results. (2) High DNA load was predictive of CVC removal and i.v. treatment duration, before blood culture results became available at 48 hours after sampling. (3) There was limited evidence that antibiotic lock treatment reduces the risk of recurrent CVC-associated infection or CVC removal (pooled relative risk 0.7, 95% CI 0.47 to 1.05), but prophylactic use of antimicrobial locks halved the risk of bloodstream infection (pooled incidence rate ratio 0.43, 95% CI 0.36 to 0.51). Contrary to this, the national survey of paediatric oncology centres found that locks are being used for treatment rather than prevention and that problems related to the formulation of lock solutions currently impede a shift to their prophylactic use in children. (4) Most i.v. treatment days would be saved by early stopping of treatment for children at low risk of infection. LIMITATIONS: The accuracy study was limited primarily by the lack of an adequate reference standard, and the main limitation of the series of systematic reviews was the poor quality of included studies and lack of randomised controlled trials of CVC removal or antimicrobial locks for treatment of infection. CONCLUSIONS: There is strong evidence to support the use of antimicrobial locks for prevention of CVC-associated infection; however, few of these studies involved children with cancer. The analysis does not support routine bacterial DNA testing on admission to detect CVC-associated infection, but repeated testing (as a marker of microbial load) should be evaluated in high-risk groups. Further research should determine the effectiveness of antibiotic locks for treating CVC-associated infection. TRIAL REGISTRATION: Current Controlled Trials ISRCTN68138140. FUNDING: This project was funded by the NIHR Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 15, No. 7. See the HTA programme website for further project information.