Expression of eukaryotic-like protein in the microbiome of sponges.

Eukaryotic-like proteins (ELPs) are classes of proteins that are found in prokaryotes, but have a likely evolutionary origin in eukaryotes. ELPs have been postulated to mediate host-microbiome interactions. Recent work has discovered that prokaryotic symbionts of sponges contain abundant and diverse genes for ELPs, which could modulate interactions with their filter-feeding and phagocytic host. However, the extent to which these ELP genes are actually used and expressed by the symbionts is poorly understood. Here, we use metatranscriptomics to investigate ELP expression in the microbiomes of three different sponges (Cymbastella concentrica, Scopalina sp. and Tedania anhelens). We developed a workflow with optimized rRNA removal and in silico subtraction of host sequences to obtain a reliable symbiont metatranscriptome. This showed that between 1.3% and 2.3% of all symbiont transcripts contain genes for ELPs. Two classes of ELPs (cadherin and tetratricopeptide repeats) were abundantly expressed in the C. concentrica and Scopalina sp. microbiomes, while ankyrin repeat ELPs were predominant in the T. anhelens metatranscriptome. Comparison with transcripts that do not encode ELPs indicated a constitutive expression of ELPs across a range of bacterial and archaeal symbionts. Expressed ELPs also contained domains involved in protein secretion and/or were co-expressed with proteins involved in extracellular transport. This suggests these ELPs are likely exported, which could allow for direct interaction with the sponge. Our study shows that ELP genes in sponge symbionts represent actively expressed functions that could mediate molecular interaction between symbiosis partners.

In vitro discrimination of wound-associated bacteria by volatile compound profiling using selected ion flow tube-mass spectrometry.

AIMS: To determine if bacterial species responsible for clinically relevant wound infection produce specific volatile profiles that would allow their speciation. METHODS AND RESULTS: Selected ion flow tube-mass spectrometry (SIFT-MS) in full mass scan mode was used to analyse headspace gases produced by wound-associated bacteria grown in vitro, so as to enable identification of bacterial volatile product ion profiles in the resulting mass spectra. Applying multivariate statistical analysis (hierarchical clustering and principal component analysis) to the resultant mass spectra enabled clear speciation. Moreover, bacterial volatile product ions could be detected from artificially contaminated wound dressing material, although the pattern of product ions detected was influenced by culture conditions. CONCLUSIONS: Using selected product ions from the SIFT-MS mass spectra it is possible to discriminate wound-associated bacterial species grown under specific in vitro culture conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study have shown that wound-associated bacteria can be discriminated using volatile analysis in vitro and that bacterial volatiles can be detected from wound dressing material. This indicates that volatile analysis of wounds or dressing material to identify infecting microbes has potential and warrants further study.

Assessing the antimicrobial potential of aerosolised electrochemically activated solutions (ECAS) for reducing the microbial bio-burden on fresh food produce held under cooled or cold storage conditions.

The main aim of this study was to assess the antimicrobial efficacy of electrochemically activated fog (ECAF) for reducing the microbial bio-burden on artificially inoculated fresh produce held under cooled (cucumber and vine tomatoes) or cold (rocket and broccoli) storage conditions. The ECAF treatment (1100 ± 5 mV ORP; 50 ± 5 mg L-1 free chlorine; 2.7 ± 0.1 pH) resulted in a significant log reduction in the potential pathogen E. coli recovered from rocket (2.644 Log10 CFU g-1), broccoli (4.204 Log10 CFU g-1), cucumber (3.951 Log10 CFU g-1) and tomatoes (2.535 Log10 CFU g-1) after 5 days. ECAF treatment also resulted in a significant log reduction in potential spoilage organisms, whereby a 3.533 Log10 CFU g-1, 2.174 Log10 CFU g-1 and 1.430 Log10 CFU g-1 reduction in presumptive Pseudomonads was observed for rocket, broccoli and cucumber respectively, and a 3.527 Log10 CFU g-1 reduction in presumptive Penicillium spp. was observed for tomatoes (after 5 days). No adverse visual effects on produce were recorded. The results of this study will inform industrial scale-up trials within commercial facilities (assessing shelf-life, microbial quality and organoleptic assessment) to assess the developed ECAF technology platform within a real food processing environment.

Seasonal migration of Cnaphalocrocis medinalis (Lepidoptera: Crambidae) over the Bohai Sea in northern China.

The rice leaf roller, Cnaphalocrocis medinalis (Guenée), is a serious insect pest of rice with a strong migratory ability. Previous studies on the migration of C. medinalis were mostly carried out in tropical or subtropical regions, however, and what the pattern of seasonal movements this species exhibits in temperate regions (i.e. Northern China, where they cannot overwinter) remains unknown. Here we present data from an 11-year study of this species made by searchlight trapping on Beihuang Island (BH, 38°24'N; 120°55'E) in the centre of the Bohai Strait, which provides direct evidence that C. medinalis regularly migrates across this sea into northeastern agricultural region of China, and to take advantage of the abundant food resources there during the summer season. There was considerable seasonal variation in number of C. medinalis trapped on BH, and the migration period during 2003-2013 ranged from 72 to 122 days. Some females trapped in June and July showed a relatively higher proportion of mated and a degree of ovarian development suggesting that the migration of this species is not completely bound by the 'oogenesis-flight syndrome'. These findings revealed a new route for C. medinalis movements to and from Northeastern China, which will help us develop more effective management strategies against this pest.

Is breastfeeding in infancy predictive of child mental well-being and protective against obesity at 9 years of age?

BACKGROUND: Preventing child mental health problems and child obesity have been recognized as public health priorities. The aim of the present study was to examine whether being breastfed (at all or exclusively) in infancy was a predictor of mental well-being and protective against risk of obesity at age 9. METHODS: Cross-sectional data from a large, nationally representative cohort study in the Republic of Ireland was used (n = 8357). Data on breastfeeding were retrospectively recalled. Child mental well-being was assessed using a parent-completed Strengths and Difficulties Questionnaire (SDQ). Child's height and weight were measured using scientifically calibrated instruments. RESULTS: Logistic regression analyses indicated that, after controlling for a wide range of potential confounding variables, being breastfed in infancy was associated with a 26% (P < 0.05) reduction in the risk of an abnormal SDQ score at 9 years. Being breastfed remained a significant predictor of child mental well-being when child obesity was controlled for, indicating that being breastfed, independent of child obesity, is a predictor of child mental well-being. The results of a second logistic regression indicated that, after controlling for a wide range of potential confounding variables, being breastfed for between 11 and 25 weeks was associated with a 36% (P < 0.05) reduction in the risk of obesity at 9 years, while being breastfed for 26 weeks or longer was associated with a 48% (P < 0.01) reduction in the risk of obesity at 9 years. CONCLUSIONS: Breastfeeding in infancy may protect against both poor mental well-being and obesity in childhood.

Bacterial chondronecrosis with osteomyelitis in broilers: influence of sires and straight-run versus sex-separate rearing.

Two experiments (E1, E2) were conducted to compare the influence of sires (sire A on dam C vs. sire B on dam C) and straight-run versus sex-separate rearing on the incidence of bacterial chondronecrosis with osteomyelitis (BCO) in broilers. Fertile eggs from commercial breeder flocks were incubated and hatched at the University of Arkansas Poultry Research Hatchery. Male and female chicks were reared together (straight-run) or separately (sex-separate) in 3 × 3 m pens on litter or flat wire flooring with 65 (E1) or 60 (E2) birds per pen. Necropsies revealed lesions that are pathognomonic for BCO in ≥98% of the birds that became lame. The SigmaStat Z-test was used to compare cumulative BCO incidences through 8 wk of age. For birds reared on litter, the incidences of BCO were low regardless of cross or sex (range: 1.7 to 5.1%; P ≥ 0.6). Within a cross and sex, rearing the broilers straight-run versus sex-separate on wire flooring did not significantly affect the incidence of BCO. Significant incidences of BCO did not develop until after d 40. Males from the sire A cross developed a higher incidence of BCO than males from the sire B cross in E1 (27 vs. 17%, respectively; P = 0.009) but not in E2 (28.5 vs. 22.6%, respectively; P = 0.141). In both experiments, males from the sire A cross developed higher incidences of BCO than females from the sire B cross (27 vs. 11.9%, in E1; 28.5 vs. 14.8%, in E2). With the sexes pooled, broilers from the sire A cross consistently developed higher incidences of BCO than broilers from the sire B cross (21.4 vs. 14.9%, P = 0.005 in E1; 26.5 vs. 18.7%, P = 0.003 in E2). High susceptibilities to both femoral head (all femoral head necrosis = 66 to 85% incidences) and tibial head (all tibial head necrosis = 81 to 96% incidences) BCO lesions were demonstrated in lame birds from both sexes and crosses. This study supports a sire influence on the susceptibility of broilers to BCO. Sire lines can be chosen to reduce BCO susceptibility when broilers are grown beyond 6 wk of age.

Interannual Survival of Myotis lucifugus (Chiroptera: Vespertilionidae) near the Epicenter of White-Nose Syndrome.

Reduced populations of Myotis lucifugus (Little Brown Myotis) devastated by white-nose syndrome (WNS) persist in eastern North America. Between 2009 and 2013, we recaptured 113 marked individuals that survived between 1 and 6 winters in New England since the arrival of WNS. We also observed signs of reproductive success in 57 recaptured bats.

Comparative antimicrobial activities of aerosolized sodium hypochlorite, chlorine dioxide, and electrochemically activated solutions evaluated using a novel standardized assay.

The main aim of this study was to develop a standardized experimental assay to enable differential antimicrobial comparisons of test biocidal aerosols. This study represents the first chlorine-matched comparative assessment of the antimicrobial activities of aerosolized sodium hypochlorite, chlorine dioxide, and electrochemically activated solution (ECAS) to determine their relative abilities to decontaminate various surface-associated health care-relevant microbial challenges. Standard microbiological challenges were developed by surface-associating typed Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis spores, or a clinical methicillin-resistant S. aureus (MRSA) strain on stainless steel, polypropylene, or fabric. All test coupons were subjected to 20-min biocidal aerosols of chlorine-matched (100 ppm) sodium hypochlorite, chlorine dioxide, or ECAS within a standard aerosolization chamber using a commercial humidifier under defined conditions. Biocidal treatment type and material surface had a significant effect on the number of microorganisms recovered from various material surfaces following treatment exposure. Under the conditions of the assay, the order of antimicrobial efficacy of biocidal aerosol treatment was as follows: ECAS > chlorine dioxide > sodium hypochlorite. For all biocides, greater antimicrobial reductions were seen when treating stainless steel and fabric than when treating plastic-associated microorganisms. The experimental fogging system and assay protocol designed within this study were shown capable of differentiating the comparative efficacies of multiple chlorine-matched biocidal aerosols against a spectrum of target organisms on a range of test surface materials and would be appropriate for testing other biocidal aerosol treatments or material surfaces.

Pharmacokinetic-pharmacodynamic determinants of oseltamivir efficacy using data from phase 2 inoculation studies.

Given the limited understanding about pharmacokinetic-pharmacodynamic (PK-PD) determinants of oseltamivir efficacy, data from two phase 2 influenza virus inoculation studies were evaluated. Healthy volunteers in studies 1 and 2 were experimentally infected with influenza A/Texas (the concentration of neuraminidase inhibitor which reduced neuraminidase activity by 50% [IC(50)] = 0.18 nM) or B/Yamagata (IC(50) = 16.76 nM), respectively. In study 1, 80 subjects received 20, 100, or 200 mg of oral oseltamivir twice daily (BID), 200 mg oseltamivir once daily, or placebo for 5 days. In study 2, 60 subjects received 75 or 150 mg of oral oseltamivir BID or placebo for 5 days. Oseltamivir carboxylate (OC) (active metabolite) PK was evaluated using individual PK data and a population PK model to derive individual values for area under the concentration-time curve from 0 to 24 h (AUC(0-24)), minimum concentration of OC in plasma (C(min)), and maximum concentration of OC in plasma (C(max)). Exposure-response relationships were evaluated for continuous (area under composite symptom score curve [AUCSC], area under the viral titer curve, and peak viral titer) and time-to-event (alleviation of composite symptom scores and cessation of viral shedding) efficacy endpoints. Univariable analyses suggested the existence of intuitive and highly statistically significant relationships between OC AUC(0-24 )evaluated as a 3-group variable and AUCSC, time to alleviation of composite symptom scores, and time to cessation of viral shedding. The upper OC AUC(0-24) threshold (~14,000 ng · h/ml) was similar among these endpoints. Multivariable analyses failed to demonstrate the influence of study/strain on efficacy endpoints. These results provide the first demonstration of exposure-response relationships for efficacy for oseltamivir against influenza and suggest that OC exposures beyond those achieved with the approved oseltamivir dosing regimen will provide enhanced efficacy. The clinical applicability of these observations requires further investigation.

Population pharmacokinetic analysis of clopidogrel in healthy Jordanian subjects with emphasis optimal sampling strategy.

AIM: Clopidogrel is metabolized primarily into an inactive carboxyl metabolite (clopidogrel-IM) or to a lesser extent an active thiol metabolite. A population pharmacokinetic (PK) model was developed using NONMEM(®) to describe the time course of clopidogrel-IM in plasma and to design a sparse-sampling strategy to predict clopidogrel-IM exposures for use in characterizing anti-platelet activity. METHODS: Serial blood samples from 76 healthy Jordanian subjects administered a single 75 mg oral dose of clopidogrel were collected and assayed for clopidogrel-IM using reverse phase high performance liquid chromatography. A two-compartment (2-CMT) PK model with first-order absorption and elimination plus an absorption lag-time was evaluated, as well as a variation of this model designed to mimic enterohepatic recycling (EHC). Optimal PK sampling strategies (OSS) were determined using WinPOPT based upon collection of 3-12 post-dose samples. RESULTS: A two-compartment model with EHC provided the best fit and reduced bias in C(max) (median prediction error (PE%) of 9.58% versus 12.2%) relative to the basic two-compartment model, AUC(0-24) was similar for both models (median PE% = 1.39%). The OSS for fitting the two-compartment model with EHC required the collection of seven samples (0.25, 1, 2, 4, 5, 6 and 12 h). Reasonably unbiased and precise exposures were obtained when re-fitting this model to a reduced dataset considering only these sampling times. CONCLUSIONS: A two-compartment model considering EHC best characterized the time course of clopidogrel-IM in plasma. Use of the suggested OSS will allow for the collection of fewer PK samples when assessing clopidogrel-IM exposures.

Susceptibility of 4 commercial broiler crosses to lameness attributable to bacterial chondronecrosis with osteomyelitis.

Growing broilers on wire flooring provides an excellent experimental model for exposing susceptibility to lameness attributable to bacterial chondronecrosis with osteomyelitis (BCO). Two independent experiments (E1, E2) were designed to compare the susceptibilities of broilers from 4 commercial crosses (W, X, Y, and Z). The standard crosses (W and Y) grow rapidly at an early age, whereas high-yield crosses (X and Z) initially tend to grow more slowly. Chicks were obtained from a commercial hatchery for E1, or were hatched at the University of Arkansas Poultry Research Hatchery for E2. Males and females were reared together (E1; n = 360/cross) or separately (E2; n = 390/cross) in 3 × 3 m pens on litter or wire flooring (wire). Necropsies revealed lesions that were pathognomonic for BCO in ≥94% of the birds that became lame. The SigmaStat Z-test was used to compare cumulative lameness incidences at 8 wk of age. For birds reared on litter, lameness incidences were low and did not differ between crosses or sexes (range: 2.2 to 4.6%; P ≥ 0.6). When males were reared on wire, their lameness incidences (by cross) were E1 = 52% for W(b); 42% for X(c); 69% for Y(a), and 44% for Z(bc); E2 = 31% for W(b); 19% for X(c); 49% for Y(a); and 25% for Z(bc). For females reared on wire, the lameness incidences were E1 = 40% for W(b), 30% for X(c), 49% for Y(a), and 28% for Z(c); E2 = 16% for W; 15% for X; 16% for Y; and 15% for Z (ns). Accordingly, the hierarchical ranking for BCO susceptibility by broiler cross was X ≤ Z ≤ W < Y for males in E1 and E2, for females in E1, and for males and females pooled in E1 and E2. Standard broiler crosses developed higher incidences of lameness than high-yield crosses, implicating an association between rapid early growth and susceptibility to BCO. Rearing the females separately on wire in E2 led to uniformly low incidences of BCO, regardless of cross. Stress-mediated immunosuppression contributes to the pathogenesis of BCO; perhaps female broilers experience less social or competitive stress when reared separately from their male hatch mates.

Local formularies.

The widespread availability of authoritative guidance on prescribing from a wide variety of international and national bodies calls into question the need for additional local formulary advice. This article describes contemporary local formulary management in the United Kingdom and discusses the areas where local decision making remains valuable. Local formularies can fulfil important roles which justify their continued existence, including ensuring local ownership and acceptance of advice, rapid dissemination of information, responsiveness to local circumstances and service design, sensitivity to local pricing arrangements and close professional links with commissioners, pharmacists and prescribers.

The roles of clinical pharmacologists in formulating medicines policy locally.

Specialists in Clinical Pharmacology and Therapeutics (CPT) can add value in day-to-day NHS activities in several ways. They provide a breadth of expertise that is not organ-based or disease-specific and that is based on an intimate knowledge and understanding of the effectiveness, safety and cost-effectiveness of pharmacological interventions. More than any other professional group, they can address the growing need for undergraduate and postgraduate teaching to be based on 'thoughtful therapeutics', not just the mechanics of prescribing. CPT specialists can and do take the lead in making local and national policy decisions relating to drug usage and they should be involved in local commissioning decisions. Because of the breadth of experience embraced by CPT, many clinical pharmacologists have taken on local and national senior clinical leadership roles. CPT needs to demonstrate to the NHS, and in particular to trainees, that a CPT post in the NHS is a legitimate and rewarding career path, where they can use their hard earned CPT skills and expertise to the benefit of the NHS as a whole.

Electrochemically activated solutions: evidence for antimicrobial efficacy and applications in healthcare environments.

Due to the limitations associated with the use of existing biocidal agents, there is a need to explore new methods of disinfection to help maintain effective bioburden control, especially within the healthcare environment. The transformation of low mineral salt solutions into an activated metastable state, by electrochemical unipolar action, produces a solution containing a variety of oxidants, including hypochlorous acid, free chlorine and free radicals, known to possess antimicrobial properties. Electrochemically activated solutions (ECAS) have been shown to have broad-spectrum antimicrobial activity, and have the potential to be widely adopted within the healthcare environment due to low-cost raw material requirements and ease of production (either remotely or in situ). Numerous studies have found ECAS to be highly efficacious, as both a novel environmental decontaminant and a topical treatment agent (with low accompanying toxicity), but they are still not in widespread use, particularly within the healthcare environment. This review provides an overview of the scientific evidence for the mode of action, antimicrobial spectrum and potential healthcare-related applications of ECAS, providing an insight into these novel yet seldom utilised biocides.

A case study: The deployment of a novel in situ fluorimeter for monitoring biological contamination within the urban surface waters of Kolkata, India.

The quality and health of many of our vital freshwater systems are poor. To tackle this with ever increasing pressures from anthropogenic and climatic changes, we must improve water quality monitoring and devise and implement more appropriate water quality parameters. Recent research has highlighted the potential for Peak T fluorescence (tryptophan-like fluorescence, TLF) to monitor microbial activity in aquatic systems. The VLux TPro (Chelsea Technologies Ltd., UK), an in situ real-time fluorimeter, was deployed in different urban freshwater bodies within Kolkata (West Bengal, India) during March 2019. This study is the first to apply this technology in surface waters within a densely populated urban area. Spot-sampling was also undertaken at 13 sampling locations enabling physicochemical analysis, bacterial enumeration and determination of nutrient (nitrate and phosphate) concentrations. This case study has demonstrated the ability of an in situ fluorimeter, VLux TPro, to successfully identify both biological contamination events and potential elevated microbial activity, related to nutrient loading, in complex surface freshwaters, without the need for expensive and time-consuming laboratory analysis.

A mutation in a case of early onset narcolepsy and a generalized absence of hypocretin peptides in human narcoleptic brains.

We explored the role of hypocretins in human narcolepsy through histopathology of six narcolepsy brains and mutation screening of Hcrt, Hcrtr1 and Hcrtr2 in 74 patients of various human leukocyte antigen and family history status. One Hcrt mutation, impairing peptide trafficking and processing, was found in a single case with early onset narcolepsy. In situ hybridization of the perifornical area and peptide radioimmunoassays indicated global loss of hypocretins, without gliosis or signs of inflammation in all human cases examined. Although hypocretin loci do not contribute significantly to genetic predisposition, most cases of human narcolepsy are associated with a deficient hypocretin system.

The flight paths of honeybees recruited by the waggle dance.

In the 'dance language' of honeybees, the dancer generates a specific, coded message that describes the direction and distance from the hive of a new food source, and this message is displaced in both space and time from the dancer's discovery of that source. Karl von Frisch concluded that bees 'recruited' by this dance used the information encoded in it to guide them directly to the remote food source, and this Nobel Prize-winning discovery revealed the most sophisticated example of non-primate communication that we know of. In spite of some initial scepticism, almost all biologists are now convinced that von Frisch was correct, but what has hitherto been lacking is a quantitative description of how effectively recruits translate the code in the dance into flight to their destinations. Using harmonic radar to record the actual flight paths of recruited bees, we now provide that description.

Mycobacterium tuberculosis-specific cellular immune profiles suggest bacillary persistence decades after spontaneous cure in untreated tuberculosis.

Individuals with self-healed tuberculosis from the preantibiotic era offer a unique insight into the natural history of and protective immunity to tuberculosis. In 27 such persons whose tuberculosis self-healed >50 years earlier, circulating Mycobacterium tuberculosis antigen-specific interferon γ (IFN-γ)- and interleukin 2 (IL-2)-secreting T cells were detected ex vivo in 16 and 19 individuals, respectively. The M. tuberculosis-specific T cell cytokine profile was dominated by effector memory T cells that secrete both IFN-γ and IL-2 and included T cells that secrete only IFN-γ or IL-2, suggesting persistence of antigen secreted by viable bacilli. Of 10 individuals with no M. tuberculosis antigen-specific IFN-γ-secreting T cells detectable ex vivo, 7 had evidence of central memory T cells, consistent with clearance of infection.

Influence of climate and reproductive timing on demography of little brown myotis Myotis lucifugus.

1. Estimating variation in demographic rates, such as survival and fecundity, is important for testing life-history theory and identifying conservation and management goals. 2. We used 16 years (1993-2008) of mark-recapture data to estimate age-specific survival and breeding probabilities of the little brown myotis Myotis lucifugus LeConte in southern New Hampshire, USA. Using Kendall & Nichols' (1995) full-likelihood approach of the robust design to account for temporary emigration, we tested whether survival and breeding propensity is influenced by regional weather patterns and timing of reproduction. 3. Our results demonstrate that adult female survival of M. lucifugus ranged from 0.63 (95% CL = 0.56, 0.68) to 0.90 (95% CL = 0.77, 0.94), and was highest in wet years with high cumulative summer precipitation. First-year survival [range: 0.23 (95% CL = 0.14, 0.35) to 0.46 (95% CL = 0.34, 0.57)] was considerably lower than adult survival and depended on pup date of birth, such that young born earlier in the summer (c. late May) had a significantly higher probability of surviving their first year than young born later in the summer (c. mid-July). Similarly, the probability of young females returning to the maternity colony to breed in the summer following their birth year was higher for individuals born earlier in the summer [range: 0.23 (95% CL = 0.08, 0.50) to 0.53 (95% CL = 0.30, 0.75)]. 4. The positive influence of early parturition on 1st-year survival and breeding propensity demonstrates significant fitness benefits to reproductive timing in this temperate insectivorous bat. 5. Climatic factors can have important consequences for population dynamics of temperate bats, which may be negatively affected by summer drying patterns associated with global climate change. 6. Understanding long-term demographic trends will be important in the face of a novel disease phenomenon (White-Nose Syndrome) that is associated with massive mortalities in hibernating bat species, including M. lucifugus, in the northeastern United States.