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Equine herpesvirus 1 (EHV-1) is an Alphaherpesvirus infecting not only horses but also other equid and non-equid mammals. It can cause respiratory distress, stillbirth and neonatal death, abortion, and neurological disease. The different forms of disease induced by EHV-1 infection can have dramatic consequences on the equine industry, and thus the virus represents a great challenge for the equine and scientific community. This report describes the progress of a major EHV-1 outbreak that took place in Normandy in 2009, during which the three forms of disease were observed. A collection of EHV-1 strains isolated in France and Belgium from 2012 to 2018 were subsequently genetically analysed in order to characterise EHV-1 strain circulation. The open reading frame 30 (ORF30) non-neuropathogenic associated mutation A 2254 was the most represented among 148 samples analysed in this study. ORF30 was also sequenced for 14 strains and compared to previously published sequences. Finally, a more global phylogenetic approach was performed based on a recently described Multilocus Sequence Typing (MLST) method. French and Belgian strains were clustered with known strains isolated in United Kingdom and Ireland, with no correlation between the phylogeny and the time of collection or location. This new MLST approach could be a tool to help understand epidemics in stud farms., Competing Interests: The authors declare no conflict of interest.

Fidaxomicin is a macrocyclic antibiotic licensed for treating Clostridium difficile infection (CDI). In the UK, fidaxomicin is often reserved for severe CDI or recurrences. At Queen Elizabeth Hospital Birmingham, all courses of fidaxomicin during 2017/2018 were reviewed. Thirty-eight patients received fidaxomicin, of which 64% responded to treatment when fidaxomicin was given during the first episode of mild CDI. Conversely, all patients with recurrent CDI failed treatment with fidaxomicin. There were mixed results for the use of fidaxomicin for severe CDI, with only 42% of patients responding. These results suggest that fidaxomicin is best suited as a treatment for mild CDI during a patient's first episode., (Copyright © 2019 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

In August 2015, Malaysia experienced an outbreak of acute respiratory disease in racehorses. Clinical signs observed were consistent with equine influenza (EI) infection. The index cases were horses recently imported from New Zealand. Rapid control measures, including temporary cancellation of racing, were implemented to minimize the impact of the outbreak. By November, the disease outbreak was resolved, and movement restrictions were lifted. The aim of this study was to confirm the clinical diagnosis and characterize the causal virus. A pan-reactive influenza type A real-time RT-PCR was used for confirmatory diagnosis. Antigenic characterization by haemagglutinin inhibition using a panel of specific ferret antisera indicated that the causal virus belonged to clade 1 of the H3N8 Florida sub-lineage. The genetic characterization was achieved by the whole genome sequencing of positive nasal swabs from clinically affected animals. Pylogenetic analysis of the haemagglutinin (HA) and neuraminidase (NA) genes demonstrated ≥99% homology with several EI strains that had recently circulated in the USA and Japan. The antigenic and genetic characterization did not indicate that the current World Organisation for Animal Health (OIE) recommendations for EI vaccine composition required modification., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Background and Aim: Teat disinfection is an important tool in reducing the incidence of bovine mastitis. Identifying the potential mastitis-causing bacterial species in milk can be the first step in choosing the correct teat disinfectant product. The objective of this study was to screen commercial teat disinfectants for inhibition against mastitis-associated bacteria isolated from various types of milk samples., Materials and Methods: Twelve commercially available teat disinfectant products were tested, against 12 mastitis-associated bacteria strains isolated from bulk tank milk samples and bacterial strains isolated from clinical (n=2) and subclinical (n=3) quarter foremilk samples using the disk diffusion method., Results: There was a significant variation (7-30 mm) in bacterial inhibition between teat disinfection products, with products containing a lactic acid combination (with chlorhexidine or salicylic acid) resulting in the greatest levels of bacterial inhibition against all tested bacteria (p<0.05)., Conclusion: In this study, combined ingredients in teat disinfection products had greater levels of bacterial inhibition than when the ingredients were used individually. The disk diffusion assay is a suitable screening method to effectively differentiate the bacterial inhibition of different teat disinfectant products.

Background: Pseudomonas aeruginosa is a ubiquitous and important opportunistic pathogen in immunocompromised or critically ill patients. Nosocomial P. aeruginosa outbreaks have been associated with hospital water sources., Aim: To describe engineering interventions to minimize contamination of water outlets and the subsequent clinical impact., Methods: New tap outlets were fitted at selected outlets across the intensive care unit (ICU). Laboratory testing demonstrated that, following artificial contamination with P. aeruginosa, these taps could be effectively decontaminated using a thermal washer-disinfector. Water samples were collected weekly from new outlets on the ICU over an eight-month period and tested for the enumeration of P. aeruginosa via membrane filtration. Surveillance of P. aeruginosa from clinical specimens was routinely undertaken., Findings: Prior to the interventions, water sampling on ICU indicated that 30% of the outlets were positive for P. aeruginosa at any one time, and whole genome sequencing data suggested at least 30% transmission from water to patient. Since their installation, weekly sampling of the new tap outlets has been negative for P. aeruginosa, and the number of P. aeruginosa clinical isolates has fallen by 50%., Conclusion: Installation and maintenance of tap outlets free of P. aeruginosa can substantially reduce the number of P. aeruginosa clinical isolates in an ICU., (Copyright © 2018 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

The use of pre- and post-milking teat disinfectants can reduce teat bacterial load and aid in the collection of high-quality milk. The objective of this study was to compare the reduction in bacteria populations on teat skin after the application of different commercial teat disinfectant products. Ten teat disinfectant products were applied to the teats of 10 Holstein-Friesian cows. One cow received one teat disinfectant product at each sampling point before cluster application for milking. A composite swab sample was taken of the 4 teats of each cow before and after teat disinfectant application. Swab samples were placed on three different selective agars to enumerate bacterial counts of staphylococcal, streptococcal and coliforms isolates on teat skin. Staphylococcal isolates were the most prominent bacterial group recovered on teat swabs (49%), followed by streptococcal (36%) and coliform (15%) isolates before the application of disinfectant. The average bacterial reductions on teat skin were shown to be 76%, 73% and 60% for staphylococcal, streptococcal and coliform isolates, respectively. All of the teat disinfectant products tested reduced teat bacterial load for all three bacterial groups. Product 4 containing 0.6% w/w diamine was the most effective against bacterial populations of staphylococcal and streptococcal isolates on teat skin with a reduction of 90% and 94%, respectively. Whereas product 10, which contained 0.5% w/w iodine, resulted in the highest reduction in coliforms on teat skin with a reduction of 91%. Results from this study suggest that specific bacterial population loads on teats can be reduced using different teat disinfectant formulations.

The purpose of this study was to examine associations between the Walk Score and physical activity in young, overweight/obese urban women. Project Health included 45 White or African American women (BMI 31.5±3.9 kg/m2; age 26.5±4.6 years; 62% African American) living in the Boston area. An accelerometer estimated steps/day and mins/day in light physical activity (100-2019 counts-per-minute) and moderate-to-vigorous-physical activity (≥2020 cpm). Walk Score was used to estimate the walkability of home address by analyzing proximity to nearby amenities. General linear regression models estimated associations between total Walk Score and physical activity (light physical activity, moderate-to-vigorous-physical activity, steps, total activity counts, METs), adjusting for body mass index, age, race/ethnicity, seasonality, wear time, employment and student status. For physical activity variables that had significant associations with Walk Score (steps/day and steps/min), regression models were estimated for Walk Score sub-scores (parks, grocery, errands, shopping, dining/drinking, culture/entertainment and schools). Logistic regression models estimated the odds of meeting the guidelines for steps (≥10,000/day) and moderate-to-vigorous-physical activity (≥150mins MVPA/week) based on Walk Score. Participants had a Walk Score of 63.9±26.4, took 14,143±3,934 steps/day, and spent 206.2±66.0 mins/day in light physical activity and 46.7±17.5 mins/day in moderate-to-vigorous- physical activity. Walk Score was significantly and positively associated with steps/day (β = 51.4, p = 0.01) and steps/min (β = 0.06, p = 0.009) but was not associated with mins/day of light physical activity, moderate-to-vigorous-physical activity, total activity counts or METs. Parks, grocery, errands, shopping, dining/drinking, and culture/entertainment Walk Score sub-scores were significantly associated with steps and steps/min (all p<0.05), but not significantly associated for schools. Participants who lived in higher Walk Score neighborhoods were more likely to meet the step guidelines (OR, 95% CI: 1.59, 1.04-2.99) and moderate-to-vigorous-physical activity guidelines (1.63, 1.06-3.02), respectively, per 10-unit increase in Walk Score. These results indicate that living in a more walkable neighborhood may support walking behavior in young, urban-dwelling overweight/obese women and provide further evidence for the expanded use of urban planning and transportation policies to improve the walkability of urban neighborhoods., Competing Interests: The authors have declared no competing interests exist.

Multiple locus typing based on sequencing heterologous regions in 26 open reading frames (ORFs) of equine herpesvirus 1 (EHV-1) strains Ab4 and V592 was used to characterise 272 EHV-1 isolates from 238 outbreaks of abortion, respiratory or neurological disease over a 28-year period. The analysis grouped the 272 viruses into at least 10 of the 13 unique long region (U L ) clades previously recognised. Viruses from the same outbreak had identical multi-locus profiles. Sequencing of the ORF68 region of EHV-1 isolates from 222 outbreaks established a divergence into seven groups and network analysis demonstrated that Irish genotypes were not geographically restricted but clustered with viruses from all over the world. Multi-locus analysis proved a more comprehensive method of strain typing than ORF68 sequencing. It was demonstrated that when interpreted in combination with epidemiological data, this type of analysis has a potential role in tracking virus between premises and therefore in the implementation of targeted control measures. Viruses from 31 of 238 outbreaks analysed had the proposed ORF30 G2254/D752 neuropathogenic marker. There was a statistically significant association between viruses of the G2254/D752 genotype and both neurological disease and hypervirulence as defined by outbreaks involving multiple abortion or neurological cases. The association of neurological disease in those with the G2254/D752 genotype was estimated as 27 times greater than in those with the A2254/N752 genotype.

Background: Mycobacterium tuberculosis is a major health burden worldwide. The disease may present as an individual case, community outbreak, or more rarely as a nosocomial outbreak. Even in countries with a low prevalence such as the UK, tuberculosis (TB) presents a risk to healthcare workers (HCWs)., Aim: To report an outbreak which manifested 12 months after a patient with pulmonary tuberculosis was admitted to Queen Elizabeth Hospital Birmingham., Methods: We present the epidemiological and outbreak investigations; the role of whole genome sequencing (WGS) in identifying the outbreak and control measures to prevent further outbreaks., Findings: Subsequent to a diagnosis of open TB in a patient, transmission was confirmed in one HCW who had active TB; HCWs with latent TB infection (LTBI) were also identified among seven HCW contacts of the index patient. Of note, all the LBTI patients had other risk factors for TB. Routine use of WGS identified the outbreak link between the index patient and the HCW with active TB disease, and informed our investigations., Conclusion: Exposure most likely occurred during an aerosol-generating procedure (AGP) which was done in accordance with national guidance at that time without using respiratory protection. Enhanced control measures were implemented following the outbreak., (Copyright © 2018 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Endogenous peptides regulate plant immunity and growth. Systemin, a peptide specific to the Solanaceae, is known for its functions in plant responses to insect herbivory and pathogen infections. Here, we describe the identification of the tomato ( Solanum lycopersicum ) PEPR1/2 ORTHOLOG RECEPTOR-LIKE KINASE1 (PORK1) as the TOMATO PROTEIN KINASE1b (TPK1b) interacting protein and demonstrate its biological functions in systemin signaling and tomato immune responses. Tomato PORK1 RNA interference (RNAi) plants with significantly reduced PORK1 expression showed increased susceptibility to tobacco hornworm ( Manduca sexta ), reduced seedling growth sensitivity to the systemin peptide, and compromised systemin-mediated resistance to Botrytis cinerea. Systemin-induced expression of Proteinase Inhibitor II ( PI-II ), a classical marker for systemin signaling, was abrogated in PORK1 RNAi plants. Similarly, in response to systemin and wounding, the expression of jasmonate pathway genes was attenuated in PORK1 RNAi plants. TPK1b, a key regulator of tomato defense against B. cinerea and M. sexta , was phosphorylated by PORK1. Interestingly, wounding- and systemin-induced phosphorylation of TPK1b was attenuated when PORK1 expression was suppressed. Our data suggest that resistance to B. cinerea and M. sexta is dependent on PORK1-mediated responses to systemin and subsequent phosphorylation of TPK1b. Altogether, PORK1 regulates tomato systemin, wounding, and immune responses., (© 2018 American Society of Plant Biologists. All rights reserved.)

Background: In 2013, there was an outbreak of acute respiratory disease in racehorses in Turkey. The clinical signs were consistent with equine influenza (EI)., Objective: The aim was to confirm the cause of the outbreak and characterise the causal virus., Methods: A pan-reactive influenza type A real-time RT-PCR and a rapid antigen detection kit were used for confirmatory diagnosis of equine influenza virus (EIV). Immunological susceptibility to EIV was examined using single radial haemolysis and ELISA. Antigenic characterisation was completed by haemagglutinin inhibition using a panel of specific ferret antisera. Genetic characterisation was achieved by whole-genome sequencing using segment-specific primers with M13 tags., Results: A H3N8 EIV of the Florida clade 2 sublineage (FC2) was confirmed as the causal agent. The index cases were unvaccinated and immunologically susceptible. Phylogenetic analysis of the HA1 and NA genes demonstrated that A/equine/Ankara/1/2013 clustered with the FC2 strains circulating in Europe. Antigenic characterisation confirmed the FC2 classification and demonstrated the absence of significant drift. Whole-genome sequencing indicated that A/equine/Ankara/1/2013 is most closely related to the viruses described as the 179 group based on the substitution I179V in HA1, for example A/equine/East Renfrewshire/2/2011, A/equine/Cambremer/1/2012 and A/equine/Saone et Loire/1/2015. The greatest diversity was observed in the NS1 segment and the polymerase complex., Conclusions: The first recorded outbreak of EI in Turkey was caused by an FC2 virus closely related to viruses circulating in Europe. Antigenic and genetic characterisation gave no indication that the current OIE recommendations for EI vaccine composition require modification., (© 2017 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.)

Equine influenza (EI) outbreaks occurred on 19 premises in Ireland during 2014. Disease affected thoroughbred (TB) and non-TB horses/ponies on a variety of premises including four racing yards. Initial clinical signs presented on 16 premises within a two-month period. Extensive field investigations were undertaken, and the diagnostic effectiveness of a TaqMan RT-PCR assay was demonstrated in regularly-vaccinated and sub-clinically-affected horses. Epidemiological data and repeat clinical samples were collected from 305 horses, of which 40% were reported as clinically affected, 39% were identified as confirmed cases and 11% were sub-clinically affected. Multivariable analysis demonstrated a significant association between clinical signs and age, vaccination status and number of vaccine doses received. Vaccine breakdown was identified in 31% of horses with up to date vaccination records. This included 27 horses in four different racing yards. Genetic and antigenic analysis identified causal viruses as belonging to Clade 2 of the Florida sublineage (FCL2). At the time of this study, no commercially available EI vaccine in Ireland had been updated in line with World Organisation for Animal Health (OIE) recommendations to include a FCL2 virus. The findings of this study highlight the potential ease with which EI can spread among partially immune equine populations.

Chronic lymphocytic leukemia (CLL) remains the most prevalent form of leukemia in the Western world, with no cure to date. Ongoing and essential research into this heterogeneous disease has led to a number of new treatment options becoming available to CLL patients in the past decade. The present review presents the recent developments in the field of CLL treatment, with the main focus on elderly patients and CLL patients with coexisting comorbidities. The review discusses the current treatment regimens that provide the most promising outcomes for patients in this subgroup, with a number of important clinical trials summarized. These clinical trials, which have investigated promising single-agent therapies or combination therapies, are discussed, with an emphasis on the efficacy and tolerability for patients aged ≥ 65 years. Also, the misrepresentation of the true CLL population in many clinical trials and the need for better guidelines for participant inclusion criteria to provide a more realistic and accurate study population are noted., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Equid herpesvirus 8 (EHV-8), formerly known as asinine herpesvirus 3, is an alphaherpesvirus that is closely related to equid herpesviruses 1 and 9 (EHV-1 and EHV-9). The pathogenesis of EHV-8 is relatively little studied and to date has only been associated with respiratory disease in donkeys in Australia and horses in China. A single EHV-8 genome sequence has been generated for strain Wh in China, but is apparently incomplete and contains frameshifts in two genes. In this study, the complete genome sequences of four EHV-8 strains isolated in Ireland between 2003 and 2015 were determined by Illumina sequencing. Two of these strains were isolated from cases of abortion in horses, and were misdiagnosed initially as EHV-1, and two were isolated from donkeys, one with neurological disease. The four genome sequences are very similar to each other, exhibiting greater than 98.4% nucleotide identity, and their phylogenetic clustering together demonstrated that genomic diversity is not dependent on the host. Comparative genomic analysis revealed 24 of the 76 predicted protein sequences are completely conserved among the Irish EHV-8 strains. Evolutionary comparisons indicate that EHV-8 is phylogenetically closer to EHV-9 than it is to EHV-1. In summary, the first complete genome sequences of EHV-8 isolates from two host species over a twelve year period are reported. The current study suggests that EHV-8 can cause abortion in horses. The potential threat of EHV-8 to the horse industry and the possibility that donkeys may act as reservoirs of infection warrant further investigation.

Background: There has been a dramatic increase in the number of hospital-acquired infections, which is linked to the pandemic of multidrug resistance. Clinical environments provide an ideal reservoir for the growth, proliferation, and transmission of pathogenic organisms, including bacterial and yeast species. Consequently, the need for improved, effective disinfectants is of paramount importance., Methods: Studies were conducted to assess the efficacy of chemical disinfectants-peracetic acid and triameen-on microbial strains. Testing included the assessment of antimicrobial and antisporicidal activity of disinfection solutions performed on a range of clinical isolates that pose a high risk for patient morbidity in clinical settings., Results: Both chemical disinfectants successfully inactivated all test strains, with peracetic acid showing a greater level of antimicrobial activity. Escherichia coli proved most susceptible when assessed by the Kirby disk diffusion, suspension, and medical suspension assays with the greatest reduction in cell viability achieved. Antibiotic-resistant Enterococcus and Staphylococcus aureus strains showed greatest resistance to both disinfectants., Discussion and Conclusions: Test chemicals show potential to act as intermediate-level disinfectants inactivating vegetative microorganisms and bacterial spores on clinically relevant strains where they show potential as a preventative measure in relation to nosocomial infections., (Copyright © 2018 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Mycobacterium avium subspecies paratuberculosis is a bacterial parasite and the causative agent of paratuberculosis, a disease predominately found in cattle and sheep. Infection with this microorganism results in substantial farming economic losses and animal morbidity. The link between infection with this pathogen and human disease has been theorised for many years with Crohn's disease being one of many suspected resultant conditions. Mycobacterium avium may be spread from animal to human hosts by water and foodborne transmission routes, where the foodborne route of exposure represents a significant risk for susceptible populations, namely children and the immune-compromised. Following colonisation of the host, the parasitic organism evades the host immune system by use of molecular mimicry, displaying peptide sequences similar to that of the host cells causing a disruption of self-verses non self-recognition. Theoretically, this failure to recognise the invading organism as distinct from host cells may result in numerous autoimmune conditions. Here, the author presents current information assessing the link between numerous diseases states in humans such inflammatory bowel disease, Type 1 diabetes, rheumatoid arthritis, Hashimoto\'s thyroiditis, multiple sclerosis and autism following infection with Mycobacterium avium paratuberculosis . The possibility of zoonotic transmission of the organism and its significant risk to public health safety as a consequence is also discussed.

Pseudomonas aeruginosa is an important nosocomial pathogen widely colonizing hospital water supplies. The Department of Health (England) Health Technical Memorandum (HTM) 04-01 addresses the risk posed by recommending water-testing in augmented care areas including outpatient haemodialysis. We discuss how two teaching hospitals independently reviewed the risk to outpatient haemodialysis patients, drawing the same conclusion. The highest number of infection episodes with P. aeruginosa was observed in critical care followed by burns and haematology, with the lowest in haemodialysis. Based on these results, we suggest that water sampling should be undertaken in areas such as critical care, burns, and haematology, but not in outpatient haemodialysis., (Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Amyloid fibril formation occurs from a wide range of peptides and proteins and is typically associated with a loss of protein function and/or a gain of toxic function, as the native structure of the protein undergoes major alteration to form a cross β-sheet array. It is now well recognised that some amyloid fibrils have a biological function, which has led to increased interest in the potential that these so-called functional amyloids may either retain the function of the native protein, or gain function upon adopting a fibrillar structure. Herein, we investigate the molecular chaperone ability of α-crystallin, the predominant eye lens protein which is composed of two related subunits αA- and αB-crystallin, and its capacity to retain and even enhance its chaperone activity after forming aggregate structures under conditions of thermal and chemical stress. We demonstrate that both eye lens α-crystallin and αB-crystallin (which is also found extensively outside the lens) retain, to a significant degree, their molecular chaperone activity under conditions of structural change, including after formation into amyloid fibrils and amorphous aggregates. The results can be related directly to the effects of aging on the structure and chaperone function of α-crystallin in the eye lens, particularly its ability to prevent crystallin protein aggregation and hence lens opacification associated with cataract formation., Competing Interests: The authors declare no conflict of interest. The funding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

The role of heater-cooler units (HCUs) in the transmission of Mycobacterium chimaera during open heart surgery has been recognized since 2013. Subsequent investigations uncovered a remarkable global outbreak reflecting the wide distribution of implicated devices. HCUs are an essential component of cardiopulmonary bypass operations and their withdrawal would severely affect capacity for life-saving cardiac surgery. However, studies have demonstrated that many HCUs are contaminated with a wide range of micro-organisms, including M. chimaera and complex biofilms. Whole genome sequencing of M. chimaera isolates recovered from one manufacturer's HCUs, worldwide, has demonstrated a high level of genetic similarity, for which the most plausible hypothesis is a point source contamination of the devices. Dissemination of bioaerosols through breaches in the HCU water tanks is the most likely route of transmission and airborne bacteria have been shown to have reached the surgical field even with the use of ultraclean theatre ventilation. Controlling the microbiological quality of the water circulating in HCUs and reducing biofilm formation has been a major challenge for many hospitals. However, enhanced decontamination strategies have been recommended by manufacturers, and, although they are not always effective in eradicating M. chimaera from HCUs, UK hospitals have not reported any new cases of M. chimaera infection since implementing these mitigation strategies. Water safety groups in hospitals should be aware that water in medical devices such as HCUs may act as a vector in the transmission of potentially fatal water-borne infections., (Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Background: Hepatitis C virus (HCV) infection is a major health burden worldwide. A patient with no history of HCV infection while on a renal unit was found to seroconvert to HCV., Aim: To report the use of sequencing to postulate how transmission of HCV occurred in a healthcare setting, and how this guided our outbreak investigation., Findings: Based on infection control inspections the transmission event was surmised to be due to ward environmental contamination with blood and subsequent inoculation from intravenous interventions on the patient acquiring HCV. We discuss the interventions put in place in response to the outbreak investigation findings., Conclusion: Sequencing of healthcare-acquired HCV infections should be undertaken as routine practice in outbreak investigations., (Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

The aim was to carry out a phylogenetic study of bovine viral diarrhoea viruses (BVDV) circulating in Irish cattle herds from 2011 to 2014. Three hundred and twenty five viruses from 267 herds were subtyped by nucleotide sequence analysis of the 5'UTR and/or the N pro regions. All viruses investigated in this study belonged to species BVDV-1 with BVDV-1a as the prominent subtype (97%). Subtypes BVDV-1b, BVDV-1d and BVDV-1e were also identified for the first time in Ireland. Pairwise alignments of 225 viruses with complete sequences for the 5'UTR and the N pro regions were performed to determine a low conflict threshold for virus strain demarcation. One hundred and seventy seven unique virus strains were identified. The study revealed significant levels of herd specific clustering of strains but no geographical or temporal clustering. Similar virus strains were identified in different counties, provinces and years indicating the potential to investigate the epidemiology of the disease by combining sequence analysis with animal movement data., (Copyright © 2016. Published by Elsevier Ltd.)

The interactions that occur during HIV Pr55Gag oligomerization and genomic RNA packaging are essential elements that facilitate HIV assembly. However, mechanistic details of these interactions are not clearly defined. Here, we overcome previous limitations in producing large quantities of full-length recombinant Pr55Gag that is required for isothermal titration calorimetry (ITC) studies, and we have revealed the thermodynamic properties of HIV assembly for the first time. Thermodynamic analysis showed that the binding between RNA and HIV Pr55Gag is an energetically favourable reaction (ΔG<0) that is further enhanced by the oligomerization of Pr55Gag. The change in enthalpy (ΔH) widens sequentially from: (1) Pr55Gag-Psi RNA binding during HIV genome selection; to (2) Pr55Gag-Guanosine Uridine (GU)-containing RNA binding in cytoplasm/plasma membrane; and then to (3) Pr55Gag-Adenosine(A)-containing RNA binding in immature HIV. These data imply the stepwise increments of heat being released during HIV biogenesis may help to facilitate the process of viral assembly. By mimicking the interactions between A-containing RNA and oligomeric Pr55Gag in immature HIV, it was noted that a p6 domain truncated Pr50Gag Δp6 is less efficient than full-length Pr55Gag in this thermodynamic process. These data suggest a potential unknown role of p6 in Pr55Gag-Pr55Gag oligomerization and/or Pr55Gag-RNA interaction during HIV assembly. Our data provide direct evidence on how nucleic acid sequences and the oligomeric state of Pr55Gag regulate HIV assembly.

Pseudomonas aeruginosa is an important nosocomial pathogen, colonizing hospital water supplies including taps and sinks. We report a cluster of P. aeruginosa acquisitions during a period of five months from tap water to patients occupying the same burns single room in a critical care unit. Pseudomonas aeruginosa cultured from clinical isolates from four different patients was indistinguishable from water strains by pulsed-field gel electrophoresis. Water outlets in critical care may be a source of P. aeruginosa despite following the national guidance, and updated guidance and improved control measures are needed to reduce the risks of transmission to patients., (Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Unmodified racemic sites on heterogeneous chiral catalysts reduce their overall enantioselectivity, but this effect is mitigated in the Orito reaction (methyl pyruvate (MP) hydrogenation to methyl lactate) by an increased hydrogenation reactivity. Here, this effect is explored on a R-1-(1-naphthyl)ethylamine (NEA)-modified Pd(111) model catalyst where temperature-programmed desorption experiments reveal that NEA accelerates the rates of both MP hydrogenation and H/D exchange. NEA+MP docking complexes are imaged using scanning tunnelling microscopy supplemented by density functional theory calculations to allow the most stable docking complexes to be identified. The results show that diastereomeric interactions between NEA and MP occur predominantly by binding of the C=C of the enol tautomer of MP to the surface, while simultaneously optimizing C=O····H2N hydrogen-bonding interactions. The combination of chiral-NEA driven diastereomeric docking with a tautomeric preference enhances the hydrogenation activity since C=C bonds hydrogenate more easily than C=O bonds thus providing a rationale for the catalytic observations.

Background: Non-tuberculosis mycobacteria such as Mycobacterium chimaera are found widely in hospital water systems. Invasive M. chimaera infections have recently been attributed to heater-cooler units (HCUs) of cardiopulmonary bypass equipment., Aim: To assess the extent of microbiological contamination within the HCUs and to inform decontamination strategies for reducing the microbial load., Methods: Water samples taken from HCUs used at University Hospitals Birmingham for cardiopulmonary bypass surgery were sampled to determine the number of micro-organisms by membrane filtration. Various decontamination processes were used throughout the study, all based on the manufacturer's guidance., Findings: Total viable counts >300cfu per 100mL containing a wide variety of micro-organisms were obtained from water inside the HCUs. Working with the manufacturers, we significantly reduced the microbial load of the water within the HCUs by removing the internal tubing soiled with biofilm followed by a weekly decontamination regimen with peracetic acid., Conclusion: A decontamination cycle including an initial replacement of internal tubing with weekly microbiological water samples is required to maintain the water quality within HCUs at an acceptable level., (Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Over the last decade, carbapenemase-producing organisms (CPOs) have spread worldwide, becoming a major public health concern. This article reports the authors' experience in dealing with a burns patient infected with CPOs, and the decontamination methods employed to render a burns shock room safe for re-use. The shock room was cleaned after being vacated, but environmental sampling cultured multiple CPOs. A second decontamination was undertaken comprising a detergent, steam and hypochlorite clean followed by hydrogen peroxide misting, and no CPOs were cultured after subsequent environmental sampling. A burns patient harbouring CPOs contaminates the surroundings heavily, so standard cleaning is insufficient to reduce the environmental bioburden., (Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Objectives: The detailed structure of brain-derived Aβ amyloid fibrils is unknown. To approach this issue, we investigate the molecular architecture of Aβ(1-40) fibrils grown in either human cerebrospinal fluid solution, in chemically simple phosphate buffer in vitro or extracted from a cell culture model of Aβ amyloid plaque formation., Methods: We have used hydrogen-deuterium exchange (HX) combined with nuclear magnetic resonance, transmission electron microscopy, seeding experiments both in vitro and in cell culture as well as several other spectroscopic measurements to compare the morphology and residue-specific conformation of these different Aβ fibrils., Results and Conclusions: Our data reveal that, despite considerable variations in morphology, the spectroscopic properties and the pattern of slowly exchanging backbone amides are closely similar in the fibrils investigated. This finding implies that a fundamentally conserved molecular architecture of Aβ peptide fold is common to Aβ fibrils.

In occupying an intermediate trophic position, herbivorous insects serve a vital link between plants at the base of the food chain and parasitoids at the top. Although these herbivore-mediated indirect plant-parasitoid interactions are well-documented, new studies have uncovered previously undescribed mechanisms that are fundamentally changing how we view tri-trophic relationships. In this review we highlight recent advances in this field focusing on both plant-driven and parasitoid-driven outcomes that flow up and down the trophic web, respectively. From the bottom-up, plant metabolites can impact parasitoid success by altering host immune function; however, few have considered the potential effects of other plant defense strategies such as tolerance on parasitoid ecology and behavior. From the top-down, parasitoids have long been considered plant bodyguards, but in reality the consequences of parasitism for herbivory rates and induction of plant defensive chemistry are far more complicated with cascading effects on community-level interactions., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Research into more rapid and effective means of disinfecting water has become necessary due to the recognition that not all pathogenic species are being removed by chemical means. There is an extent of research highlighting the benefits of pulsed light for the disinfection of water. This study aims to determine the ability of a real time polymerase chain reaction assay to evaluate microbial inactivation of pulsed light treated cells. Findings show that pulsed light is a more rapid means of inactivating test species than standard UV lamp systems. A linear relationship between cell number and polymerase chain reaction amplification was obtained. A difference in threshold value (Ct) of approximately 4 (p ≤ 0.05) was obtained for DNA amplification following the addition of the dye for pulsed ultrviolet (PUV)-treated Bacillus cells. Membrane protein leakage proved an effective means of determining membrane damage for both Bacillus and E. coli test species following PUV treatment. This membrane damage was not evident for cells exposed to low pressure ultraviolet (LPUV). Findings describe suggest that PUV treatment induced a viable but nonculturable state in treated cells.

Objective: This review discusses the relevance of the National Institute of Mental Health (NIMH) Research Domain Criteria (RDoC) to clinical research in child and adolescent psychiatry., Method: We summarize the characteristics of the NIMH RDoC project and then provide examples of RDoC designs that are of relevance to clinical investigators in child and adolescent psychiatry. The final section addresses questions regarding the impact of RDoC on clinical care., Results: RDoC encourages investigators to investigate psychopathology dimensionally: greater or lesser degrees of healthy/adapted functioning of neurobiological, cognitive, and behavioral processes (constructs) that cut across current diagnostic categories. Elucidation of the developmental components of RDoC constructs is needed to ensure they are fully validated. Integrating RDoC approaches into clinical research of child and adolescent psychopathology is contributing to our understanding of development as an aspect of the heterogeneity within DSM disorders and commonalities across seemingly disparate disorders. Continued efforts promise to also explain the processes that lead to mental illness in at-risk populations., Conclusion: Incorporating an RDoC approach in clinical research in child and adolescent psychiatry promises to be a fruitful avenue of research into the root causes and manifestations of mental illness, which will eventually lead to more precise treatments. Although the long-term aspiration of RDoC is to help reduce the burden of suffering for those with mental illnesses, it is not intended to be used for practical clinical purposes at this early stage., (Published by Elsevier Inc.)

The presence of pathogenic organisms namely parasite species and bacteria in biofilms in veterinary settings, is a public health concern in relation to human and animal exposure. Veterinary clinics represent a significant risk factor for the transfer of pathogens from housed animals to humans, especially in cases of wound infection and the shedding of faecal matter. This study aims to provide a means of detecting veterinary relevant parasite species in bacterial biofilms, and to provide a means of disinfecting these biofilms. A real time PCR assay was utilized to detect parasite DNA in Bacillus cereus biofilms on stainless steel and PVC surfaces. Results show that both Cryptosporidium and Giardia attach to biofilms in large numbers (100-1000 oo/cysts) in as little as 72 hours. Pulsed light successfully inactivated all test species (Listeria, Salmonella, Bacillus, Escherichia) in planktonic and biofilm form with an increase in inactivation for every increase in UV dose.

Viruses are often thought to have static structure, and they only remodel after the viruses have entered target cells. Here, we detected a size expansion of virus particles prior to viral entry using cryo-electron microscopy (cryo-EM) and single molecule fluorescence imaging. HIV expanded both under cell-free conditions with soluble receptor CD4 (sCD4) targeting the CD4 binding site on the HIV-1 envelope protein (Env) and when HIV binds to receptor on cellular membrane. We have shown that the HIV Env is needed to facilitate receptor induced virus size expansions, showing that the 'lynchpin' for size expansion is highly specific. We demonstrate that the size expansion required maturation of HIV and an internal capsid core with wild type stability, suggesting that different HIV compartments are linked and are involved in remodelling. Our work reveals a previously unknown event in HIV entry, and we propose that this pre-entry priming process enables HIV particles to facilitate the subsequent steps in infection., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Children living in poverty are at high risk for delays in development of language and behavior and they experience a discrepancy in diagnosis and access to intervention services. This gap is partially caused by barriers in access as well as traits that are specific to each child and family. The Identification of Neurodevelopmental Disabilities in Underserved Children using Telehealth (INvesT) trial is a novel intervention approach that was specifically designed to address these barriers., Aims: The INvesT trial has three primary aims: 1) to reduce the age of identification of neurodevelopmental disability for high-risk, low-income children. 2) To validate the INvesT protocol as a service delivery model that will decrease age of identifications of neurodevelopmental disability for high-risk, low-income children; and 3) to identify important child-specific factors, family-specific factors, and environmental factors that impact feasibility and success of the INvesT trial for high-risk, low-income children., Methods: The INvesT trial is an open-label, double-blinded, placebo-controlled multi-level study that includes telehealth risk assessment, telehealth screening, traditional full assessment, and follow through to enrollment in early intervention. The trial is conducted in partnership with an urban community health clinic that largely serves a low-income patient population., Conclusions: The results of the INvesT trial will provide evidence for the use of a telehealth service delivery model to improve access to care for neurodevelopmental disabilities for high-risk, low-income children., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Microorganisms are naturally found as biofilm communities more than planktonic free-floating cells; however, planktonic culture remains the current model for microbiological studies, such as disinfection techniques. The presence of fungal biofilms in the clinical setting has a negative impact on patient mortality, as Candida biofilms have proved to be resistant to biocides in numerous in vitro studies; however, there is limited information on the effect of pulsed light on sessile communities. Here we report on the use of pulsed UV light for the effective inactivation of clinically relevant Candida species. Fungal biofilms were grown by use of a CDC reactor on clinically relevant surfaces. Following a maximal 72 h formation period, the densely populated biofilms were exposed to pulsed light at varying fluences to determine biofilm sensitivity to pulsed-light inactivation. The results were then compared to planktonic cell inactivation. High levels of inactivation of C. albicans and C. parapsilosis biofilms were achieved with pulsed light for both 48 and 72 h biofilm structures. The findings suggest that pulsed light has the potential to provide a means of surface decontamination, subsequently reducing the risk of infection to patients. The research described herein deals with an important aspect of disease prevention and public health., (Copyright © 2015 John Wiley & Sons, Ltd.)

The use of ultraviolet (UV) light for water disinfection has become increasingly popular due to on-going issues with drinking water and public health. Pulsed UV light has proved to be an effective form of inactivating a range of pathogens including parasite species. However, there are limited data available on the use of pulsed UV light for the disinfection of flowing water in the absence or presence of inorganic contaminants commonly found in water sources. Here, we report on the inactivation of test species including Bacillus endospores following pulsed UV treatment as a flow through system. Significant levels of inactivation were obtained for both retention times tested. The presence of inorganic contaminants iron and/or manganese did affect the rate of disinfection, predominantly resulting in an increase in the levels of inactivation at certain UV doses. The findings of this study suggest that pulsed UV light may provide a method of water disinfection as it successfully inactivated bacterial cells and bacterial endospores in the absence and presence of inorganic contaminants.

Aggregation of amyloid-beta (Aβ) peptide is the major event underlying neuronal damage in Alzheimer's disease (AD). Specific lipids and their homeostasis play important roles in this and other neurodegenerative disorders. The complex interplay between the lipids and the generation, clearance or deposition of Aβ has been intensively investigated and is reviewed in this chapter. Membrane lipids can have an important influence on the biogenesis of Aβ from its precursor protein. In particular, increased cholesterol in the plasma membrane augments Aβ generation and shows a strong positive correlation with AD progression. Furthermore, apolipoprotein E, which transports cholesterol in the cerebrospinal fluid and is known to interact with Aβ or compete with it for the lipoprotein receptor binding, significantly influences Aβ clearance in an isoform-specific manner and is the major genetic risk factor for AD. Aβ is an amphiphilic peptide that interacts with various lipids, proteins and their assemblies, which can lead to variation in Aβ aggregation in vitro and in vivo. Upon interaction with the lipid raft components, such as cholesterol, gangliosides and phospholipids, Aβ can aggregate on the cell membrane and thereby disrupt it, perhaps by forming channel-like pores. This leads to perturbed cellular calcium homeostasis, suggesting that Aβ-lipid interactions at the cell membrane probably trigger the neurotoxic cascade in AD. Here, we overview the roles of specific lipids, lipid assemblies and apolipoprotein E in Aβ processing, clearance and aggregation, and discuss the contribution of these factors to the neurotoxicity in AD.

Research into alternative methods of disinfecting water and wastewater has proven necessary due to the emergence of chlorine-resistant organisms and the disinfection byproducts associated with chlorine use. The use of UV light to inactivate microbial species has proven effective, however; standard UV lamps have proven to be less effective in their ability to inactivate parasites and bacterial endospores in water treatment settings. Pulsed UV (PUV) light may potentially provide a novel alternative to water and wastewater disinfection. Research outlined in this study assesses the potential of a novel PUV system for the rapid and reproducible inactivation of a range of test species including Bacillus endospores. In comparison to standard low-pressure (LP) UV lamps, this PUV system provided significantly higher levels of inactivation for all test species. Furthermore, there was a remarkable decrease in time needed to obtain significant inactivation rates following treatment with PUV compared to LP-UV. With the PUV system, a 70-second treatment time (7.65 μJ/cm2) resulted in similar inactivation rates of Bacillus endospores to that of the LP-UV inactivation of their vegetative counterpart. Also, at PUV doses exceeding 4.32 J/cm2, there was not a significant difference in the PUV inactivation of Bacillus endospores in the absence or presence of 10 ppm organic matter. However, the presence of organic matter resulted in a significant reduction in microbial inactivation for all treatment doses using the LP-UV system. The findings of this study suggest that PUV technology may provide a rapid effective method for the disinfection of water and wastewater.

Second generation biofuel development is increasingly reliant on the recombinant expression of cellulases. Designing or identifying successful expression systems is thus of preeminent importance to industrial progress in the field. Recombinant production of cellulases has been performed using a wide range of expression systems in bacteria, yeasts and plants. In a number of these systems, particularly when using bacteria and plants, significant challenges have been experienced in expressing full-length proteins or proteins at high yield. Further difficulties have been encountered in designing recombinant systems for surface-display of cellulases and for use in consolidated bioprocessing in bacteria and yeast. For establishing cellulase expression in plants, various strategies are utilized to overcome problems, such as the auto-hydrolysis of developing plant cell walls. In this review, we investigate the major challenges, as well as the major advances made to date in the recombinant expression of cellulases across the commonly used bacterial, plant and yeast systems. We review some of the critical aspects to be considered for industrial-scale cellulase production.

The structure of the 1-D oligomer chains that form on a Au(111) surface following adsorption of 1,4-phenylene diisocyanide (PDI) is explored using reflection-absorption infrared spectroscopy and scanning tunneling microscopy (STM). The experimental work is complemented by first-principles density functional theory calculations, which indicate that the previously proposed gold-PDI oligomer chains in which the PDI molecule bridged gold adatoms are thermodynamically stable. In addition, the calculated vibrational modes for this structure are in excellent agreement with the experimental infrared data. The linkage of the PDI units by gold adatoms is confirmed by comparing STM images collected as a function of tip bias with images for the calculated structure by the Bardeen method.