An in vitro investigation of the survival and/or growth of Campylobacter jejuni in broiler digestate from different feed types.

Campylobacter spp. is the leading cause of bacterial gastroenteritis worldwide and poultry are the primary reservoir. The aim of this study was to investigate the survival and/or growth of Campylobacter jejuni NCTC 11168 in broiler digestate prepared from commercial starter, grower and finisher feed formulations. Bolton broth and digestates were prepared, inoculated with C. jejuni NCTC 11168 (approximately 3 log10 CFU per ml) and incubated under microaerobic conditions at 42°C for 24 h. Samples were taken at t = 0 (immediately after inoculation) and every 3 h thereafter, serially diluted and plated onto mCCDA. Campylobacter jejuni grew as expected in Bolton broth (control) reaching the early stationary phase after approximately 15 h. In contrast, although bacterial concentrations were maintained for at least 9 h, none of the feed digestates supported the growth of C. jejuni, which were not detected after 15 h. It is suggested that the nutrients available in the feed digestates are not enough to support C. jejuni growth and that additional factors may be at play in the avian gastrointestinal tract.

The psychological impact of Stevens-Johnson syndrome and toxic epidermal necrolysis on patients' lives: a Critically Appraised Topic.

CLINICAL SCENARIO: A 65-year-old man presented with a 12-h history of deteriorating rash. Two weeks previously he had completed a course of neoadjuvant chemotherapy for ductal carcinoma of the breast. On examination there were bullae, widespread atypical targetoid lesions and 15% epidermal detachment. There was no mucosal involvement on presentation, but subsequently it did evolve. Skin biopsy showed subepidermal blistering with epidermal necrosis. This confirmed our clinical diagnosis of overlap Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN). On transfer to intensive care he was anxious and fearful. MANAGEMENT QUESTION: What are the psychological impacts of SJS/TEN on this man's life? BACKGROUND: SJS and TEN have devastating outcomes for those affected. OBJECTIVES: To conduct a Critically Appraised Topic to (i) analyse existing research related to the psychological impact of SJS and TEN and (ii) apply the results to the clinical scenario. METHODS: Seven electronic databases were searched for publications focusing on the psychological impact of SJS/TEN on adults over 18 years of age. RESULTS: Six studies met the inclusion criteria. Healthcare practitioners' (HCPs') lack of information around the disorder was highlighted. Patients experienced undue stress and fear. Some patients had symptoms aligned to post-traumatic stress disorder (PTSD), anxiety and depression. DISCUSSION AND RECOMMENDATION: The evidence suggests that SJS and TEN impact psychologically on patients' lives. Education of HCPs, to address their lack of awareness and information on SJS/TEN, should facilitate their capacity to provide information and support to patients, thereby reducing patient anxiety. On discharge, a follow-up appointment with relevant HCPs to reduce the possibility of PTSD occurring should be considered.

Antimicrobial resistance of Campylobacter isolates recovered from broilers in the Republic of Ireland in 2017 and 2018: an update.

1. Campylobacteriosis is the leading cause of human bacterial gastroenteritis. Broilers are considered the most important source of human Campylobacter infection. In the 2008 European baseline survey Ireland had a 98% prevalence of campylobacter-contaminated broiler carcases. 2. Randomly-selected Campylobacter isolates (296 C. jejuni, 54 C. coli) recovered in 2017 and 2018, from Irish broiler neck skin and caeca were tested for their resistance to tetracycline, erythromycin, gentamicin, ciprofloxacin, nalidixic acid and streptomycin. 3. Overall, 45% of the Campylobacter spp. isolates tested were resistant to at least one antimicrobial. Tetracycline resistance (38%) was most prevalent in C. jejuni, followed by ciprofloxacin and nalidixic acid resistance (29%). In C. coli, resistance to ciprofloxacin and nalidixic acid (26%) was most prevalent followed by resistance to tetracycline (13%). Gentamicin resistance was undetected and resistance to streptomycin was low for C. jejuni (1%) and C. coli (4%). All C. jejuni isolates examined were erythromycin-sensitive, while 9% of C. coli isolates were erythromycin-resistant. Three multidrug-resistant C. coli isolates were recovered. 4. While antibiotic resistance rates were somewhat similar to figures reported nationally over the past 20 years, the prevalence of tetracycline resistance in C. jejuni has increased. The persistence of substantial ciprofloxacin resistance in the Irish broiler population was noteworthy, despite fluoroquinolones having been banned for growth promotion in Europe since 2006.

Diversity of Listeria monocytogenes strains isolated from Agaricus bisporus mushroom production.

AIMS: The aims of this study were to characterize the genetic diversity of Listeria monocytogenes isolates obtained from commercial mushroom production, to establish the persistence, recontamination and the risk of cross-contamination from the working environment to the final products, creating awareness about the presence of L. monocytogenes thus helping to prevent the possibility of cross-contamination. METHODS AND RESULTS: From an extensive analysis of commercial mushroom production, analysed with BS EN ISO 11290-1:1996/Amd 1:2004 and BS EN ISO 11290-2:1998/Amd 1:2004, 279 L. monocytogenes isolates were obtained. All of the isolates were characterized by pulsed-field gel electrophoresis, species PCR and serogroup PCR. All the isolates were confirmed as L. monocytogenes; 30·1% were serogroup 1/2b-3b-7, 40·8% were serogroup 1/2a-3a and 29·1% were serogroup 4b-4d-4e. There were 77 pulsotypes from the 279 isolates, 40 of the pulsotypes had only one strain and 37 had two or more strains, indicating great diversity in the isolates. CONCLUSIONS: The high genetic diversity is indicative of the fact that current hygiene practices are successful at removing L. monocytogenes but that recontamination of the production environment is frequent. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained are very valuable in creating awareness of L. monocytogenes in mushroom production and for the improvement of hygiene practices.

The UVB photoreceptor UVR8 mediates accumulation of UV-absorbing pigments, but not changes in plant morphology, under outdoor conditions.

UVB radiation is biologically active; in plants, it can induce a range of molecular, biochemical, morphological and developmental responses. Although much progress has been made in elucidating UVB perception and signalling pathways under controlled laboratory conditions, understanding of the adaptive, ecological role of UVB responses is still very limited. In this study, we looked at the functional role of UVR8 under outdoor light conditions, by studying growth, photosynthetic competence and accumulation of UV absorbing pigments in a mutant lacking functional UVR8 protein. It was found that the influence of UVB on morphology is restricted to summer and is independent of UVR8. In contrast, UVB had an effect on the content of UV-absorbing pigments and the maximal efficiency of photosystem II of photosynthesis in the uvr8-1 mutant throughout the year. It is concluded that the UVR8 photoreceptor plays a role throughout the year, in the temperate climate zone, even when UVB levels are relatively low.

Inhibition of Listeria monocytogenes biofilms by bacteriocin-producing bacteria isolated from mushroom substrate.

AIMS: This study was designed to investigate the ability of naturally occurring bacteria isolated from mushroom substrate to prevent biofilm formation by Listeria monocytogenes or to remove existing biofilms in mushroom production facilities. METHODS AND RESULTS: It is generally recognized that L. monocytogenes forms biofilms that can facilitate its survival in food-processing environments. Eleven bacteriocin-producing isolates were identified and the bacteriocins characterized based on heat and enzyme inactivation studies. Further characterization was undertaken by MALDI-TOF mass spectrometry, PCR and sequencing. Production of nisin Z (by Lactococcus lactis isolates), subtilomycin (by Bacillus subtilis isolates) and lichenicidin (by Bacillus licheniformis and Bacillus sonorensis isolates) was detected. In co-culture with L. monocytogenes, the bacteriocin-producing strains could prevent biofilm formation and reduce pre-formed biofilms. CONCLUSIONS: Mushroom substrate can be a source of bacteriocin-producing bacteria that can antagonize L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlight the potential of bacteriocin-producing strains from mushroom substrate to reduce L. monocytogenes biofilm in food production environments, contributing to a reduction in the risk of food contamination from the environment.

Control of Zymoseptoria tritici cause of septoria tritici blotch of wheat using antifungal Lactobacillus strains.

AIMS: This study explored an effective biological control agent based on lactic acid bacteria culture or culture supernatant, which was effective against fungicide-resistant Zymoseptoria tritici, which causes septoria tritici blotch (STB). METHODS AND RESULTS: Three lactic acid bacteria strains which exhibited broad antifungal activity were investigated for their potential to control Z. tritici. Plate assays, liquid culture growth inhibition assays and STB biocontrol seedling tests were employed. Lactobacillus brevis JJ2P and Lactobacillus reuteri R2 caused significant fungal inhibition as observed by large mycelium clearing on modified MRS agar. Cell-free culture supernatants of Lact. brevis JJ2P and Lact. reuteri R2 showed antifungal activity against Z. tritici, as observed by mycelial radial growth inhibition and liquid culture growth inhibition. Cell-free supernatants of these anti-Z. tritici LAB strains were assessed in vivo for their abilities to inhibit STB development in seedling tests. Lact. brevis JJ2P was capable of inhibiting disease development and significantly reduced the diseased leaf area covered with pycnidia. CONCLUSIONS: Biological control accomplished by beneficial micro-organisms such as Lact. brevis JJ2P may represent an alternative control strategy for reducing STB. SIGNIFICANCE AND IMPACT OF THE STUDY: Globally, STB is regarded as one of the most important diseases of wheat. Control of Z. tritici is heavily reliant on fungicide application. The recent emergence of resistance or reduced sensitivity to fungicides among Z. tritici populations has urgently called for the development of new control strategies.

Investigation into the prevalence, persistence and antibiotic resistance profiles of staphylococci isolated from euro currency.

AIMS: The study set out to sample € 10 banknotes for the presence of coagulase-positive staphylococci (CPS) such as Staphylococcus aureus (S. aureus) and coagulase-negative staphylococci (CoNS) in Southern Ireland, to assess the levels of antibiotic resistance among those isolated, and determine the persistence of S. aureus on € 10 banknotes and € 2 coins. METHODS AND RESULTS: We report that 97% of € 10 banknotes screened (n = 155) harboured multiple species of staphylococci. From the generated bank of strains, a total of 150 representative staphylococci isolates were used for further study, 71 were CPS and 79 were CoNS. Of these, we found that 62% of the staphylococci demonstrated resistance to at least one of the first-line antibiotics (52.11% of CPS isolates and 76.71% of the CoNS isolates). Resistance to multiple antibiotics was seen in 31.18% of the resistant isolates. In relation to persistence studies, S. aureus was shown to remain viable on euro banknotes and coins for significant periods (on average, 19.33 days on € 10 banknotes and 16.67 days on € 2 coins) as determined using bioluminescence. CONCLUSIONS: We advocate the expansion of antibiotic surveillance programs, with a view to tracking/monitoring antibiotic resistance dissemination among environmental contaminants. Additionally, we propose that 'cashless transactions' should be encouraged in high-risk environments such as hospitals and healthcare settings, as well as stricter infection controls. SIGNIFICANCE AND IMPACT OF THE STUDY: Although it is accepted that circulating currency has the potential to harbour disease-causing pathogens, studies investigating prevalence and persistence of such pathogens on euro currency are virtually nonexistent. In an attempt to rectify this, we examined the prevalence of staphylococci on € 10 banknotes in Ireland and reported relatively high levels of antibiotic resistance among the isolates. Furthermore, we have established the persistence of S. aureus on euro currency for the first time.

Barley malt wort fermentation by exopolysaccharide-forming Weissella cibaria MG1 for the production of a novel beverage.

AIMS: The growing interest of governments and industry in developing healthy and natural alternative foods and beverages that will fulfil the consumer drive towards a healthy lifestyle and clean-label, natural diet has led to an increase in traditional lactic acid bacteria fermentation research. In particular, this research aims to address the organoleptic modulation of beverages using in situ-produced bacterial polysaccharides. METHODS AND RESULTS: Weissella cibaria MG1 is capable of producing exopolysaccharides (dextran) and oligosaccharides (glucooligosaccharides) during sucrose-supplemented barley-malt-derived wort fermentation. Up to 36·4 g l(-1) of dextran was produced in an optimized system, which improved the rheological profile of the resulting fermentate. Additionally, small amounts of organic acids were formed, and ethanol remained below 0·5% (v/v), the threshold volume for a potential health claim designation. CONCLUSIONS: The results suggest that the cereal fermentate produced by W. cibaria MG1 could be potentially used for the production of a range of novel, nutritious and functional beverages. SIGNIFICANCE AND IMPACT OF THE STUDY: Using conventional raw materials and traditional processes, novel LAB-fermented beverages can be produced representing an innovative mechanism towards fulfilling the aim to decrease government and personal costs as well as potentially ameliorating consumer lifestyle regarding dietary-related disease.

Discovery of an opportunistic starfish pathogen, Orchitophrya stellarum, in captive blue crabs, Callinectes sapidus.

Histophagous scuticociliate infections were discovered in blue crabs, Callinectes sapidus, held in research facilities at the Virginia Institute of Marine Science. Ciliates were observed infecting every tissue examined including the gills, heart, muscle, hepatopancreas, and epidermis. Hemolymph smears and histological tissue sections indicated a morphological similarity to Mesanophrys chesapeakensis, the only recorded histophagous ciliate infecting blue crabs. However, subsequent analysis of the ribosomal ITS region (ITS1-5.8S-ITS2) of the ciliate indicated the parasite was Orchitophrya stellarum, a parasitic ciliate previously reported infecting sea stars from Europe, Australia, and North America. A simple Polymerase Chain Reaction (PCR-RFLP) assay was developed to detect and differentiate between O. stellarum and M. chesapeakensis. Its application confirmed the presence of O. stellarum infecting blue crabs held in an additional research facility in Maryland. For growth studies, cultures of O. stellarum grew optimally on 10% blue crab serum in crustacean saline held at 10-20°C. A field survey of blue crabs collected during the winters of 2011-2012 and sea stars (Asterias forbesi) during the winter of 2010 from the Chesapeake Bay and eastern shore of Virginia did not identify additional infected individuals.

Host response to EBV infection in X-linked lymphoproliferative disease results from mutations in an SH2-domain encoding gene.

X-linked lymphoproliferative syndrome (XLP or Duncan disease) is characterized by extreme sensitivity to Epstein-Barr virus (EBV), resulting in a complex phenotype manifested by severe or fatal infectious mononucleosis, acquired hypogammaglobulinemia and malignant lymphoma. We have identified a gene, SH2D1A, that is mutated in XLP patients and encodes a novel protein composed of a single SH2 domain. SH2D1A is expressed in many tissues involved in the immune system. The identification of SH2D1A will allow the determination of its mechanism of action as a possible regulator of the EBV-induced immune response.

Nurses' preferred end-of-life treatment choices in five countries.

BACKGROUND: Previous research has focused on physician's perspectives of end-of-life (EOL) decision making as well as patient and family EOL decision making. There is a lack of research pertaining to the EOL treatment preferences of nurses and especially nurses working in a variety of care settings. AIM: The aim of this study was to compare nurses' EOL treatment preferences in Hong Kong, Ireland, Israel, Italy and the USA. METHODS: A comparative descriptive design was used with a convenience sample of nurses (n = 1089). A survey questionnaire using EOL hypothetical clinical case scenarios was used to collect data between June 2011 and July 2012. RESULTS: Nurses in every country consistently chose a more aggressive option for patients than for themselves or for a parent. The treatment preferences of nurses varied from country to country. Lack of knowledge of patients' wishes and duty of care were the main influencing factors on treatment preferences. STUDY LIMITATIONS: The study was limited to the hypothetical nature of the scenarios; however, the study highlights numerous future research questions. CONCLUSIONS: This study is the first to examine and compare nurses' preferred EOL treatment choices in five countries from three different continents. The findings of this study raise several important questions for healthcare researchers, for policy development, and highlight the need for further international collaboration.

Prevention of Staphylococcus aureus biofilm formation and reduction in established biofilm density using a combination of phage K and modified derivatives.

AIMS: To investigate the ability of a mixture of phage K and six of its modified derivatives to prevent biofilm formation by Staphylococcus aureus and also to reduce the established biofilm density. METHODS AND RESULTS: The bioluminescence-producing Staph. aureus Xen29 strain was used in the study, and incubation of this strain in static microtitre plates at 37°C for 48 h confirmed its strong biofilm-forming capacity. Subsequently, removal of established biofilms of Staph. aureus Xen29 with the high-titre phage combination was investigated over time periods of 24 h, 48 h and 72 h. Results suggested that these biofilms were eliminated in a time-dependent manner, with biofilm biomass reduction significantly greater after 72 h than after 24-48 h. In addition, initial challenge of Staph. aureus Xen29 with the phage cocktail resulted in the complete inhibition of biofilm formation over a 48-h period with no appearance of phage resistance. CONCLUSIONS: In general, our findings demonstrate the potential use of a modified phage combination for the prevention and successful treatment of Staph. aureus biofilms, which are implicated in several antibiotic-resistant infections. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the first use of phage K for the successful removal and prevention of biofilms of Staph. aureus.

Emergence of group B Streptococcus serotype IV in women of child-bearing age in Ireland.

This study determined the carriage rate and serotype distribution of group B Streptococcus (GBS) in women of child-bearing age in the southern region of Ireland. A total of 2000 vaginal swabs collected in two periods in 2004 and 2006 were examined and revealed a GBS carriage rate of 16·1%. Serotyping of isolates showed that serotypes Ia, II, III, IV, and V were the most prevalent. A high prevalence of serotype IV was found, increasing from 7·6% to 15·2% between 2004 and 2006. Random amplified polymorphic DNA analysis demonstrated considerable genetic heterogeneity in the serotype IV isolates. This serotype should be considered for inclusion in potential vaccines for use in Ireland.

Inhibition of growth of Trichophyton tonsurans by Lactobacillus reuteri.

AIM: The aims of this study were to identify antifungal lactic acid bacteria (LAB) and characterize their activity against the dermatophyte Trichophyton tonsurans. METHODS AND RESULTS: A total of 165 different LAB were isolated and initially screened for anti-Penicillium expansum activity. Five strains, which exhibited strong inhibitory activity, were then tested against the dermatophyte T. tonsurans DSM12285, where they also caused inhibition as observed by large fungal clearing on agar surface. The strongest inhibition was seen with Lactobacillus reuteri R2. When freeze-dried cell-free supernatant powder from this strain was incorporated in culture medium at concentrations >1%, growth of fungal colony was inhibited. Conidia germination was also inhibited under these conditions as determined by microscopy. The anti-T. tonsurans activity of Lact. reuteri R2 was not affected neither by heat treatment nor by proteolytic treatment using pronase E and proteinase K, indicating that the responsible agent(s) were nonproteinaceous in nature. CONCLUSIONS: Lactobacillus reuteri R2 was identified as having strong inhibitory activity against the dermatophyte T. tonsurans DSMZ12285. SIGNIFICANCE AND IMPACT OF THE STUDY: LAB are naturally associated with many foods and are well recognized for their biopreservative properties. The use of these and/or their products may well provide alternative safe approaches for the inhibition of dermatophytic fungi.

Characterization of the staphylococcal bacteriophage lysin CHAP(K).

AIMS: To develop an efficient purification strategy for the bacteriophage lysin CHAP(K) . To evaluate its antibacterial spectrum(,) enzymatic properties, optimal reaction conditions and lytic activity against live Staphlyococcus aureus. METHODS AND RESULTS: Recombinant CHAP(K) was purified to homogeneity by cation exchange chromatography, with yields of up to 10 mg from 1 l of Escherichia coli culture. The lytic spectrum of CHAP(K) includes all staphylococcal species and also members of the genera Micrcococcus, Streptococcus, Nesterenkonia, Arthrobacter, Leuconostoc and Carnobacterium. The enzyme was active from pH 6 to 11 with an optimum activity at pH 9, from 5 to 40°C, with an optimum activity at 15°C. When cell lysis by CHAP(K) and lysostaphin was compared over a concentration range of 2·5-10 µg ml⁻¹ using live Staph. aureus for 5 min at 37°C, CHAP(K) gave rise to greater turbidity reduction indicating that it works more rapidly than lysostaphin. CONCLUSIONS: This study describes in detail the purification and characteristics of the novel phage-derived enzyme CHAP(K) demonstrating that it has excellent biochemical properties as an anti-staphylococcal agent. SIGNIFICANCE AND IMPACT OF THE STUDY: Currently, there is a need for new antimicrobial agents due to the increasing worldwide prevalence of antibiotic resistance. Our findings demonstrate the potential for development of CHAP(K) as an alternative therapeutic against pathogenic staphylococci including MRSA.

Dementia palliative care: A multi-site survey of long term care STAFF'S education needs and readiness to change.

Many people with dementia reside in long-term care, where limited staff knowledge of dementia palliative care has been identified, along with poor awareness that a palliative approach can assist in identifying unmet care needs. Evidence-based guidance in palliative care for people with dementia is available however, implementing this guidance requires staff engagement and a tailored educational approach. This pre-implementation situational analysis informed a tailored staff education intervention to support the implementation of national guidance on dementia palliative care in long term care. Using a cross-sectional study design, underpinned by the Consolidated Framework for Implementation Research, survey data were collected on site profile, staff demographics, learning needs, and readiness-to change at three residential care sites for older people in Ireland. In total, 69 staff (predominantly nurses and healthcare attendants) completed the surveys. Medication management and management of pain were the most frequently identified learning needs. Staff were confident in their ability to implement change but de-motivation and powerlessness were substantial factors as only one-third of staff were "ready for change". Staffing levels, managing risk during change and perceived reluctance in others were common barriers. These results informed an educational intervention to address the specific care context, staff learning needs and barriers to change prior to implementation.

Emergence of MRSA clone ST22 in healthy young adults in the community in the absence of risk factors.

One thousand adults aged between 18 and 35 years were investigated for nasal colonization with community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA). Each volunteer completed a questionnaire to assess the presence or absence of risk factors for hospital-acquired MRSA (HA-MRSA) carriage. All MRSA isolated were characterized by microbiological and molecular methods. A S. aureus carriage rate of 22% and a MRSA carriage rate of 0.7% were observed. Analysis of the questionnaires revealed 121 individuals with HA-MRSA risk factors. Subsequently two MRSA infections with associated risk factors were excluded from calculation of the true carriage rate and an adjusted rate of 0.57% (5/879) was established. All seven MRSA isolates expressed the genotypic profile ST22-MRSA-IV, were PVL negative, agr type 1, and differed only by their antimicrobial susceptibility patterns. ST22-MRSA-IV (EMRSA-15) has shown worldwide spread in the hospital setting but has not been previously documented in isolation in the community.

CRISPR analysis of bacteriophage-insensitive mutants (BIMs) of industrial Streptococcus thermophilus--implications for starter design.

AIMS: An efficient approach for generation of bacteriophage-insensitive mutants (BIMs) of Streptococcus thermophilus starters was described in our laboratory [Mills et al. (2007) J Microbiol Methods70, 159-164]. The aim of this study was to analyse the phage resistance mechanism responsible for BIM formation. METHODS AND RESULTS: Three clustered regularly interspaced short palindromic repeat (CRISPR) regions have been identified in Strep. thermophilus, and Strep. thermophilus can integrate novel spacers into these loci in response to phage attack. Characterization of three sets of BIMs indicated that two sets had altered CRISPR1 and/or CRISPR3 loci. A range of BIMs of yoghurt starter CSK938 were generated with the same phage in different phage challenge experiments, and each acquired unique spacer regions ranging between one and four new spacers in CRISPR1. In addition, the BIM that acquired only one new spacer in CRISPR1 also acquired an additional spacer in CRISPR3. A fourth BIM, generated with a different phage, had two spacers deleted from CRISPR1 but acquired two spacers in CRISPR3. Analysis of the Mozzarella starter CSK939 and its associated BIMs indicated that formation of second generation BIMs does not lead to increases in spacer number but to alterations in spacer regions. BIMs of an exopolysaccharide (EPS)-producing strain that lost the ability to produce EPS did not harbour an altered CRISPR, suggesting that phage sensitivity may be related to the EPS-producing phenotype. CONCLUSIONS: Acquisition/deletion of new spacers in CRISPR loci in response to phage attack generates distinctly individual variants. It also demonstrates that other modifications may be responsible for the phage resistance of Strep. thermophilus BIMs. SIGNIFICANCE AND IMPACT OF THE STUDY: Isolation of individual BIMs that have unique spacers towards the leader region of the CRISPR locus may be a very useful approach for rotation strategies with the same starter backbone. Upon phage infection, BIMs 'in reserve' can be slotted into the rotation scheme.