This prospective cross-sectional study aimed to evaluate the effects of sociodemographic factors and geographical location on fundamental management and preventative healthcare practices in horses. The owners from UK and USA were recruited via social media and surveyed in 2023. The study collected complete data sets on fundamental management (living environment, frequency of checks/visual monitoring by owner/carer, company of other equines/ other animals, diet, monitoring of body weight and body condition) together with preventive healthcare (hoof care, dental care, endoparasite control, vaccination) from 760 horse owners.The results show that employment in equine industry and education related to equine are significant determinants in fundamental management and preventive healthcare practices provided to horses. Furthermore, the effect of time working with horses (>5 vs < 5 years) has a significant impact on the provision of vaccinations and regular veterinary health assessment/examination of horses.There were no differences detected between the UK and the USA with regard to any aspects of equine management and preventive healthcare in this study.This study results have shown that equine preventive healthcare practices and fundamentals of equine management are affected by owners’ education, practical experience and not by the geographical location (the UK vs the USA).

PURPOSE OF REVIEW:Infectious mononucleosis (IM) is an infectious disease that presents clinically in only a small percentage of individuals despite almost universal infection with the causative agent. Here, we review the latest concepts in the clinical presentation, epidemiology, and host response of this disease.RECENT FINDINGS:Several recently published papers/reviews describe IM as a condition caused by one of several etiologic agents including, cytomegalovirus (HHV-5), Roseola virus (HHV-6) and Toxoplasmosis amongst others; this review focuses on IM as solely caused by the human herpes virus 4 (HHV-4). Since the initial discovery of the virus in the 1960s and its subsequent discovery as the primary etiologic agent for IM it has been associated with several human cancers and autoimmune disorders. Recent published findings show a correlation between HHV-4 and the autoimmune disorder, multiple sclerosis (MS), suggesting earlier IM could possibly act as a causative factor. Considering the important links being made with IM to so many cancers and autoimmune disorders it is surprising that a standard investigative procedure has yet to be determined for this disease. A standard approach to the investigation of IM would ensure more cases are diagnosed, particularly atypical cases, this would benefit epidemiological studies, and more immediately help practitioners distinguish viral from bacterial throat infections, enabling them to treat accordingly.SUMMARY:The understanding of the latest concepts in clinical presentation, epidemiology and host response to IM would benefit greatly from the introduction of a standard procedure for its investigation and diagnosis.

This study aimed to generate information regarding living conditions (e.g. indoors and outdoors, living space available), fundamental care (e.g. feeding and exercise) and owners' perceptions of dog's preferences (e.g. living conditions) in the urban and rural areas of Northern Ireland via a cross-sectional survey questionnaire. The responses were collected from May to August 2022, at 24 locations across Northern Ireland, including 15 agricultural shows and livestock markets and nine large supermarkets (single supermarket chain) located in an area of the show or market grounds. In all, 548 questionnaires were collected and after exclusion of questionnaires with missing or incomprehensible responses, 507 questionnaires were included in the final database. Out of 507 questionnaires, 264 respondents resided in a city while 243 respondents resided on farms. The majority of respondents from the city locations perceived their dogs as pets, while those living on farms regarded their dogs as working animals. The populations of dogs in the city locations and on the farms in this study were similar regarding the age range and numbers of the animals, but more female dogs were spayed in the city locations than on the farms. Most respondents in this study, regardless of their household location, declared that they did not monitor their animal's body weight or body condition. When feeding their dogs, the respondents from the city locations were predominantly following veterinary advice or instructions on food labels. On the other hand, the respondents from farm locations mostly reported that they fed their dogs based on 'a visual inspection of dog condition'; this type of feeding was associated with a certain type of household occupancy (more frequent in single and adults only households) and respondents' employment status (more frequent by retired and those managing the home). The living conditions of dogs in city and farm locations in this study were different, namely dogs in the city were kept predominantly indoors with access to outdoors while dogs from farm locations were kept predominantly outdoors. The dogs were reported to be walked daily for a shorter time (up to 1 h/day) in the city locations and longer on the farm locations (1-2 h/day). Regardless of household location (city versus farm) respondents believed that exercise needs depend on animal age, body condition and medical condition, that dogs need to be kept active by owners to keep them fit, that dogs cannot self-regulate the amount of food they eat daily, and finally that walking with a dog a few times a day is difficult due to other commitments. On the other hand, the respondents from farm locations more often believe that dogs can get all the exercise they need by themselves if kept outdoors and they are happier with living outdoors, while the respondents from city locations believed that dogs are happier with living indoors. In conclusion, the results of this study have shown a number of differences in basic care and perception of dogs kept in city locations and on farm locations. Further studies are required to understand the provision of health care and fulfilments of all welfare needs of the dogs kept on farms.

The rapid emergence of multidrug-resistant pathogens worldwide has raised concerns regarding the effectiveness of conventional antibiotics. This can be observed in ESKAPE pathogens, among others, whose multiple resistance mechanisms have led to a reduction in effective treatment options. Innovative strategies aimed at mitigating the incidence of antibiotic-resistant pathogens encompass the potential use of biosurfactants. These surface-active agents comprise a group of unique amphiphilic molecules of microbial origin that are capable of interacting with the lipidic components of microorganisms. Biosurfactant interactions with different surfaces can affect their hydrophobic properties and as a result, their ability to alter microorganisms’ adhesion abilities and consequent biofilm formation. Unlike synthetic surfactants, biosurfactants present low toxicity and high biodegradability and remain stable under temperature and pH extremes, making them potentially suitable for targeted use in medical and pharmaceutical applications. This review discusses the development of biosurfactants in biomedical and therapeutic uses as antimicrobial and antibiofilm agents, in addition to considering the potential synergistic effect of biosurfactants in combination with antibiotics. Furthermore, the anti-cancer and anti-viral potential of biosurfactants in relation to COVID-19 is also discussed.

Salmonella has had a long and intimate relationship with humans and continues to raise concerns for human health, but this close bond also provides opportunities for new therapeutics and treatments. Although Salmonella enterica serovar Typhi is the principal organism that comes to mind in terms of death and morbidity, it is the non-typhoidal Salmonellae that have the most health and economic implications. The developed world has had a challenging relationship with Salmonella, particularly in the UK and the EC/EU, experiencing significant Salmonella outbreaks in the 1980s and 1990s. As a consequence, the research focus was on understanding the nature of infection in food animals and on developing ways and means of controlling zoonotic infections. This led to the development of numerous model systems for the study of Salmonella both in vitro and in vivo. The introduction of vaccination has all but eliminated Salmonella in eggs and reduced perceived risk held by the general public. At the same time as Salmonella in eggs was being brought under control in the UK and EU, the danger posed by antibiotic resistance was beginning to emerge. In the past, with the efficacy of antibiotics against Gram-negative bacteria being unchallenged, there was limited focus on the threat posed by antibiotic resistance in non-typhoidal Salmonella. However, the identification of Salmonella as the first ‘multidrug-resistant’ organism, the presence of invasive non-typhoidal Salmonella in North Africa and the emergence of monophasic Salmonella enterica serovar Typhimurium across Europe, Asia and the Americas have prompted renewed interest in Salmonella research, particularly in the context of non-infectious disease, biofilm studies and antibiotic resistance. At the same time, research has continued to develop ways of taking advantage of what Salmonella offers in the way of pathogenic factors and the therapeutic and treatment applications in areas such as vaccine development, cancer therapeutics and drug delivery and the role of Salmonella in non-infectious diseases supported by developments in molecular and genomic methods.

The 21st century has seen a substantial increase in the industrial applications of glycolipid biosurfactant technology. The market value of the glycolipid class of molecules, sophorolipids, was estimated to be USD 409.84 million in 2021, with that of rhamnolipid molecules projected to reach USD 2.7 billion by 2026. In the skincare industry, sophorolipid and rhamnolipid biosurfactants have demonstrated the potential to offer a natural, sustainable, and skin-compatible alternative to synthetically derived surfactant compounds. However, there are still many barriers to the wide-scale market adoption of glycolipid technology. These barriers include low product yield (particularly for rhamnolipids) and potential pathogenicity of some native glycolipid-producing microorganisms. Additionally, the use of impure preparations and/or poorly characterised congeners as well as low-throughput methodologies in the safety and bioactivity assessment of sophorolipids and rhamnolipids challenges their increased utilisation in both academic research and skincare applications. This review considers the current trend towards the utilisation of sophorolipid and rhamnolipid biosurfactants as substitutes to synthetically derived surfactant molecules in skincare applications, the challenges associated with their application, and relevant solutions proposed by the biotechnology industry. In addition, we recommend experimental techniques/methodologies, which, if employed, could contribute significantly to increasing the acceptance of glycolipid biosurfactants for use in skincare applications while maintaining consistency in biosurfactant research outputs.

Acidic sophorolipids (Acidic SL), congeners of sophorolipid biosurfactants, offer a potential alternative to synthetic sodium lauryl ether sulphate (SLES) in skincare applications. However, major challenges associated with the laboratory-based investigations of the cytotoxic effects of Acidic SL have been the utilisation of impure and/or poorly characterised congeners as well as the use of monolayers of skin cells in in vitro assays. While the former limitation makes glycolipids less attractive for use in academic research and skincare applications, the latter does not provide an accurate representation of the in vivo human skin. The present study, therefore, for the first time, assessed the cytotoxic effects of 96% pure Acidic SL on a 3D in vitro skin model in comparison with SLES, with the aim of investigating a natural alternative to synthetic surfactants for potential use in skincare applications. The 3D in vitro skin model was colonised with Staphylococcus epidermidis for 12 h, and afterwards treated with either Acidic SL or SLES at 100 μg mL−1 for a further 12 h. Subsequently, the cytotoxic effects of Acidic SL in comparison with SLES were assessed using a combination of microbiology, molecular biology techniques, immunoassays, and histological analyses. It was demonstrated that Acidic SL had no deleterious effects on the viability of S. epidermidis, tissue morphology, filaggrin expression, and the production of inflammatory cytokines in comparison to SLES. These findings, in conjunction with the possibility to produce Acidic SL from cheaper renewable natural resources, demonstrate that Acidic SL could offer a potential sustainable alternative to synthetic surfactants.

The human herpesvirus 4 (HHV-4) is a double-stranded DNA oncogenic virus more commonly referred to by its eponym, the Epstein–Barr virus (EBV).1, 2 It is the sole causative agent of the acute illness in humans described either as infectious mononucleosis (IM) or glandular fever.3 The disease carries a high morbidity and relies on clear, timely and informative laboratory test results for accurate diagnosis. To date, there are no international guidelines on how this disease should be investigated. Challenges for the clinician arise when atypical cases present, which makes unequivocal diagnosis challenging. The distinction of viral from bacterial throat infections is particularly important as the treatment of IM patients with antibiotics can lead to the development of ampicillin rashes.4 The clinical signs and symptoms of IM overlap with several other infectious diseases and malignant states, and a prompt diagnosis negates the need for other expensive exploratory tests in cases that present with splenomegaly, lymphadenopathy or suspect haematological conditions.5 A retrospective analysis was conducted on the full blood count (FBC) and white cell differential results of 420 hospital patients who had Monospot tests (Clearview® IM II MONO) requested in the MUH over the period 2018–2019. This review aimed to evaluate the benefit of detecting tell-tale atypical lymphocytes, which appear at the acute stage of infection, as a primary means of guiding the requesting of Monospot tests. The ADVIA 2120 haematology blood count and differential analyser performed an FBC analysis on the patients' peripheral blood. Detection of a population of atypical cells present in recent or acute cases of IM was plotted as large unstained cells (LUCs) in the analyser's scattergram and the results flagged. These LUCs describe cells that fall outside normal parameters, for example atypical lymphocytes, plasma or blast cells. Follow on microscopic examination of the flagged blood samples further classified these cells as atypical lymphocytes (also called plasmacytoid or reactive lymphocytes),6, 7 see Figure 1. A strong heterophile immune response is often produced in cases of IM; this has led to the development of the Monospot test. The Monospot detects non-specific heterophile antibodies, produced as part of the direct immunologic response to the EBV infection, but which are not specific to the infectious agent itself and are demonstrable by their ability to agglutinate animal erythrocytes including horse or bovine.8 A total of 357 (85%) of the Monospot requests were made following clinical assessment; the remaining 63 (15%) were laboratory-guided requests following on from peripheral blood film examination and the detection of atypical lymphocytes. The haematology analyser flagged Blasts/Atypicals in 37 (80%) of the 46 positive Monospot results. Patient ages ranged from 5 months to 88 years with a gender distribution of 53% female and 47% male. Table 1 shows the distribution of positive Monospot results, note the incidence of patients with IM in the 30+ age group is greatly reduced. In all, 46 patient samples (11%) demonstrated positive Monospot results, and 374 patients (89%) tested negative (Table 2). Of the 46 positive Monospot results, the majority 30 (65%), fell into the category of patients who presented with atypical lymphocytes coupled with a lymphocytosis; 10 (22%) in patients with atypical lymphocytes without a lymphocytosis and 6 (13%) in patients without any atypical lymphocytes or lymphocytosis. In total, 87% of positive Monospot results had atypical lymphocytes present, with percentages ranging from 1% to 51% and an average of 20%. Although fewer laboratory-guided Monospot requests were made, their positivity rates were 4.5 times higher. The laboratory requests were guided solely by the detection of atypical lymphocytes on the peripheral blood film. Though these atypical lymphocytes are characteristic of IM, they should not be considered pathognomonic for the disease, and confirmation should only be made in conjunction with a positive heterophile antibody (Monospot) result or failing that, following confirmation with specific EBV serology.9, 10 Patients giving a positive Monospot result in conjunction with a population of atypical lymphocytes, often with an absolute lymphocytosis, were considered acute or active cases of IM. A similar haematological picture but with a negative Monospot result could signal a heterophile negative case of IM and would require specific EBV serology to confirm. Of the 273 patients without any atypical cells, six were Monospot positive—half of these results were weak positives. The possibility the test picked up residual heterophile antibodies in these cases is plausible as detectable levels can circulate in the peripheral blood from 6 weeks to a year after they first appear.11, 12 Specific EBV serology would help determine the stage of infection in these cases. In summary, the Monospot test is a quick and inexpensive way of confirming a diagnosis of IM when seen in conjunction with a tell-tale population of atypical lymphocytes. Our data show that positivity rates fall dramatically in the 30+ age group, which concurs with the findings of published seroepidemiological studies, which indicate that the seroprevalence of the virus increases with age reaching up to over 95% in the 40+ age group.13-15 Although the initial FBC analysis may show lymphocytosis, it is the peripheral blood film examination that distinguishes the atypical lymphocytes associated with the disease. Prompt detection and reporting of these cells is helpful in the investigation and diagnosis of IM. The distinction of IM from other IM-like viral infections and bacterial tonsilitis is important as it determines patient management and ensures antibiotic treatment is prescribed only where necessary. This study suggests that Monospot tests should be delayed until after the blood film has been viewed and atypical lymphocytes reported; the patient's age should also be considered given the increased EBV seroprevalence, particularly in the 40+ age group. Patrick Naughton conducted the data collection and analysis and constructed the manuscript. Michael Healy co-supervised Patrick Naughton with Brigid Lucey, both of whom reviewed the work and the drafts of the manuscript with PN. Frances Enright provided clinical guidance and reviewed drafts of the manuscript. The authors would like to acknowledge David Hegarty for expert IT guidance and Breeda Herlihy for library assistance. This work was not funded. The authors have no competing interests and declare no conflicts of interest in this study or its reporting. Full ethical approval was granted for the current study by the Cork Research Ethics Committee [ECM 4(r) 08/12/18 & ECM 4(s) 07/05/19]. This work was granted ethical approval by Cork Research Ethics Committee (CREC no ECM3(s)05/07/19). Not applicable. Not applicable. Not applicable.

Skin irritation and allergic reactions associated with the use of skincare products formulated with synthetically derived surfactants such as sodium lauryl ether sulphate (SLES) have encouraged the search for naturally derived and biocompatible alternatives. Glycolipid biosurfactants such as sophorolipids (SL) and rhamnolipids (RL) offer a potential alternative to SLES. However, most studies on the bioactive properties of microbial glycolipids were determined using their mixed congeners, resulting in significant inter-study variations. This study aims to compare the effects of highly purified SL (acidic and lactonic) and RL (mono-RL and di-RL) congeners and SLES on a spontaneously transformed human keratinocyte cell line (HaCaT cells) to assess glycolipids’ safety for potential skincare applications. Preparations of acidic SL congeners were 100% pure, lactonic SL were 100% pure, mono-RL were 96% pure, and di-RL were 97% pure. Cell viability using XTT assays, cell morphological analyses, and immunoassays revealed that microbial glycolipids have differing effects on HaCaT cells dependent on chemical structure. Compared with SLES, acidic SL and mono-RL have negligible effects on cell viability, cell morphology, and production of pro-inflammatory cytokines. Furthermore, at non-inhibitory concentrations, di-RL significantly attenuated IL-8 production and CXCL8 expression while increasing IL-1RA production and IL1RN expression in lipopolysaccharide-stimulated HaCaT cells. Although further studies would be required, these results demonstrate that as potential innocuous and bioactive compounds, microbial glycolipids could provide a substitute to synthetic surfactants in skincare formulations and perform immunopharmacological roles in topical skin infections such as psoriasis. Key points • Purified glycolipid congeners have differing effects on human keratinocytes. • Compared with SLES, acidic sophorolipids and mono-rhamnolipids have innocuous effects on keratinocytes. • Di-rhamnolipids and mono-rhamnolipids modulate cytokine production in lipopolysaccharide stimulated human keratinocytes.

A cross-sectional survey questionnaire was developed in-house to investigate pet cat owners’ beliefs and attitudes related to the fundamental care of their pet cats. The questionnaire consisted of questions which were grouped into the following sections: (i) owners’ socio-demographics; (ii) cat(s) body weight and body condition monitoring; (iii) owners’ attitudes to cats’ dietary preferences, needs and satisfaction, (iv) owners’ perceptions of their cats’ physical exercise needs and satisfaction. The sample size of 376 was estimated to be required to represent the population of the given geographical location (Belfast, NI, UK). Hard copies of the questionnaires were distributed in January and February 2019 and in total 402 completed questionnaires were collected; questionnaires which included >20% of missing or incomprehensible responses were excluded from the database, resulting in 398 questionnaires being included in the final database. The study identified a number of socio-demographic factors associated with owners’ beliefs and attitudes that directly affect care provided to pet cats, e.g., the owner’s occupation has been identified as a factor associated with owner perception of certain cats’ behaviours, e.g., a cat brushing against the owner as food requests by their animal (Chi-Square 7.711 (df1), exact p = 0.006). Furthermore, most female respondents, aged 26–67 years and in an occupation not related to animals, reported selecting cat food based on their animal preferences (Chi-Square 10.332 (df1), exact p = 0.003). In contrast, female owners in animal and veterinary occupations were significantly more likely as compared to other respondents (Chi-Square 15.228 (df1), exact p < 0.001), to select cat food based on its perceived health benefit to the cat. Analysis of the respondents’ opinions of cats’ abilities to self-regulate physical activity showed that owners age was the main differentiating determinant, i.e., cat owners over 25 years old were significantly more likely than younger adults to believe that pet cats can regulate their own physical activity to keep healthy (Chi-Square 6.313 (df1), exact p = 0.025). Furthermore, respondents’ opinions of their cat’s ability to self-regulate feed intake were mainly associated with owner’s education level (Chi-Square 6.367 (df1), exact p = 0.036). The study results indicated that the attitude and beliefs behind the fundamental care practices provided to pet cats depends on particular demographic factors, especially owners’ education and occupation.

Enterococci, which are on the WHO list of priority pathogens, are commonly encountered in hospital acquired infection and are becoming increasing significant due to the development of strains resistant to multiple antibiotics. Enterococci are also important microorganisms in the environment, and their presence is frequently used as an indicator of faecal pollution. Their success is related to their ability to survive within a broad range of habitats and the ease by which they acquire mobile genetic elements, including plasmids, from other bacteria. The enterococci are frequently present within a bacterial biofilm, which provides stability and protection to the bacterial population along with an opportunity for a variety of bacterial interactions. Enterococci can accept extrachromosomal DNA both from within its own species and from other bacterial species, and this is enhanced by the proximity of the donor and recipient strains. It is this exchange of genetic material that makes the role of biofilms such an important aspect of the success of enterococci. There remain many questions regarding the most suitable model systems to study enterococci in biofilms and regarding the transfer of genetic material including antibiotic resistance in these biofilms. This review focuses on some important aspects of biofilm in the context of horizontal gene transfer (HGT) in enterococci.

Melanomas account for 80% of skin cancer deaths. Due to the strong relationship between melanomas and U.V. radiation, sunscreens have been recommended for use as a primary preventative measure. However, there is a need for targeted, less invasive treatment strategies. Glycolipids such as sophorolipids and rhamnolipids are microbially derived biosurfactants possessing bioactive properties such as antimicrobial, immunomodulatory and anticancer effects. This study aimed to ascertain the differing effects of glycolipids on skin cells. Highly purified and fully characterized preparations of sophorolipids and rhamnolipids were used to treat spontaneously transformed human keratinocyte (HaCaT) and the human malignant melanocyte (SK-MEL-28) cell lines. Cell viability and morphological analyses revealed that glycolipids have differential effects on the skin cells dependent on their chemical structure. Lactonic sophorolipids and mono-rhamnolipids were shown to have a significantly detrimental effect on melanoma cell viability compared to healthy human keratinocytes. These glycolipids were shown to induce cell death via necrosis. Additionally, sophorolipids were shown to significantly inhibit SK-MEL-28 cell migration. These findings suggest that glycolipids could be used as bioactive agents with selective inhibitory effects. As such, glycolipids could be a substitute for synthetically derived surfactants in sunscreens to provide additional benefit and have the potential as novel anti-skin-cancer therapies.

Clostridiodes difficile (C. difficile) was ranked an “urgent threat” by the Centers for Disease Control and Prevention (CDC) in 2019. C. difficile infection (CDI) is the most common healthcare-associated infection (HAI) in the United States of America as well as the leading cause of antibiotic-associated gastrointestinal disease. C. difficile is a gram-positive, rod-shaped, spore-forming, anaerobic bacterium that causes infection of the epithelial lining of the gut. CDI occurs most commonly after disruption of the human gut microflora following the prolonged use of broad-spectrum antibiotics. However, the recurrent nature of this disease has led to the hypothesis that biofilm formation may play a role in its pathogenesis. Biofilms are sessile communities of bacteria protected from extracellular stresses by a matrix of self-produced proteins, polysaccharides, and extracellular DNA. Biofilm regulation in C. difficile is still incompletely understood, and its role in disease recurrence has yet to be fully elucidated. However, many factors have been found to influence biofilm formation in C. difficile, including motility, adhesion, and hydrophobicity of the bacterial cells. Small changes in one of these systems can greatly influence biofilm formation. Therefore, the biofilm regulatory system would need to coordinate all these systems to create optimal biofilm-forming physiology under appropriate environmental conditions. The coordination of these systems is complex and multifactorial, and any analysis must take into consideration the influences of the stress response, quorum sensing (QS), and gene regulation by second messenger molecule cyclic diguanosine monophosphate (c-di-GMP). However, the differences in biofilm-forming ability between C. difficile strains such as 630 and the “hypervirulent” strain, R20291, make it difficult to assign a “one size fits all” mechanism to biofilm regulation in C. difficile. This review seeks to consolidate published data regarding the regulation of C. difficile biofilms in order to identify gaps in knowledge and propose directions for future study.

The aim of this review was to provide an update on the complex relationship between manure application, altered pathogen levels and antibiotic resistance. This is necessary to protect health and improve the sustainability of this major farming practice in agricultural systems based on high levels of manure production. It is important to consider soil health in relation to environment and land management practices in the context of the soil microflora and the introduction of pathogens on the health of the soil microbiome. Viable pathogens in manure spread on agricultural land may be distributed by leaching, surface run-off, water source contamination and contaminated crop removal. Thus it is important to understand how multiple pathogens can persist in manures and on soil at farm-scale and how crops produced under these conditions could be a potential transfer route for zoonotic pathogens. The management of pathogen load within livestock manure is a potential mechanism for the reduction and prevention of outbreaks infection with Escherichia coli , Listeria Salmonella , and Campylobacter . The ability of Campylobacter , E. coli , Listeria and Salmonella to combat environmental stress coupled with their survival on food crops and vegetables post-harvest emphasizes the need for further study of these pathogens along with the emerging pathogen Providencia given its link to disease in the immunocompromised and its’ high levels of antibiotic resistance. The management of pathogen load within livestock manure has been widely recognized as a potential mechanism for the reduction and prevention of outbreaks infection but any studies undertaken should be considered as region specific due to the variable nature of the factors influencing pathogen content and survival in manures and soil. Mediocre soils that require nutrients could be one template for research on manure inputs and their influence on soil health and on pathogen survival on grassland and in food crops.

Enterococci and biofilm-associated infections are a growing problem worldwide, given the rise in antibiotic resistance in environmental and clinical settings. The increasing incidence of antibiotic resistance and its propagation potential within enterococcal biofilm is a concern. This requires a deeper understanding of how enterococcal biofilm develops, and how antibiotic resistance transfer takes place in these biofilms. Enterococcal biofilm assays, incorporating the study of antibiotic resistance transfer, require a system which can accommodate non-destructive, real-time experimentation. We adapted a Gene Frame® combined with fluorescence microscopy as a novel non-destructive platform to study the conjugal transfer of vancomycin resistance in an established enterococcal biofilm.A multi-purpose fluorescent in situ hybridisation (FISH) probe, in a novel application, allowed the identification of low copy number mobile elements in the biofilm. Furthermore, a Hoechst stain and ENU 1470 FISH probe identified Enterococcus faecium transconjugants by excluding Enterococcus faecalis MF06036 donors. Biofilm created with a rifampicin resistant E. faecalis (MW01105Rif) recipient had a transfer efficiency of 2.01 × 10−3; double that of the biofilm primarily created by the donor (E. faecalis MF06036). Conjugation in the mixed enterococcal biofilm was triple the efficiency of donor biofilm. Double antibiotic treatment plus lysozyme combined with live/dead imaging provided fluorescent micrographs identifying de novo enterococcal vancomycin resistant transconjugants inside the biofilm. This is a model system for the further study of antibiotic resistance transfer events in enterococci. Biofilms promote the survival of enterococci and reduce the effectiveness of drug treatment in clinical settings, hence giving enterococci an advantage. Enterococci growing in biofilms exchange traits by means of horizontal gene transfer, but currently available models make study difficult. This work goes some way to providing a non-destructive, molecular imaging-based model system for the detection of antibiotic resistance gene transfer in enterococci.

EBV is the sole causative agent of the acute illness in humans described either as infectious mononucleosis (IM), or glandular fever. IM, when not clinically silent, can present in patients with at least two of the classic triad of symptoms of fever, pharyngitis, and lymphadenopathy. Challenges for the clinician arise when atypical cases present. Early, accurate and informed laboratory test results are vital for diagnosis, appropriate treatment, and management. A key challenge for the practitioner, particularly in cases where the illness can present atypically, is distinguishing bacterial tonsillitis infections from early acute IM. The ability to draw on timely, clear, and insightful laboratory results to distinguish viral from bacterial infection is vital. Correct and prompt diagnosis of IM can help prevent the unnecessary administration of antibiotics and mitigate the need for other expensive exploratory tests in cases of IM that present with splenomegaly, lymphadenopathy, or suspect haematological conditions. Good communication between the requesting clinician and those carrying out the investigative process, and between the different laboratory departments involved, is good practice and would ultimately benefit the patient. This communication will comprehensively review the aetiology, clinical presentation, and laboratory findings in IM with a view to promoting further research and so derive a standard diagnostic algorithm of the condition.

This prospective, descriptive study assessed caring practices of pet cat and dog owners in Northern Ireland with a survey questionnaire, focusing primarily on feeding and exercising. Owners of both pet cats and dogs reported that they fed their pets either twice or three times a day (53 and 78% for cats and dogs, respectively). However, in the case of nearly 40% of pet cat owners it transpired that they fed their cat(s) as often as the animal demanded (18.8%) or that food was available all the time (20.7%). Cat owners reported that their pets had access to outdoors (84%) and were not provided with any play time (53.1%). The highest number of pet dogs (46.1%) were reported as being walked daily for less than 1 h, and the highest number (38.4%) were played with daily for less than 1 h. Both cat (60%) and dog (61%) owners perceived the body condition of their animal ‘as it should be’ for animal age/sex. However, both pet cat (76%) and dog (63%) owners reported that neither their pet’s bodyweight nor its body condition was monitored. Our results suggest not all pet cat and dog owners are aware of the fundamentals of their animals’ feeding requirements as regards health and many provide insufficient physical activity required for their animals’ health and welfare. The major strengths of the study include the size and geographic distribution of the study population allowing the generalisation of the results to other locations.

Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge Ephydatia fluviatilis supported the transfer of vancomycin resistance between two Enterococcus faecalis strains that we previously demonstrated to exhibit pheromone responsive plasmid conjugation. Microcosm experiments exposed live and dead colonies of laboratory-grown sponges to a vancomycin-resistant donor strain and a rifampicin-resistant recipient strain of Ent. faecalis. Enterococci with both resistance phenotypes were detected on double selection plates. In comparison to controls, abundance of these presumed transconjugants increased significantly in water from sponge microcosms. Homogenized suspensions of sponge cells also yielded presumed transconjugants; however, there was no significant difference between samples from live or dead sponges. Fluorescent in situ hybridization analysis of the sponge cell matrix using species-specific probes revealed the presence of enterococci clusters with cells adjacent to each other. The results demonstrated that sponge colonies can support the horizontal transfer of antimicrobial resistance although the mechanism underlying this process, such as binding of the bacteria to the sponge collagen matrix, has yet to be fully elucidated.

Enterococci are robust gram-positive bacteria that are found in a variety of surroundings and that cause a significant number of healthcare-associated infections. The genus possesses a high-efficiency pheromone-responsive plasmid (PRP) transfer system for genetic exchange that allows antimicrobial-resistance determinants to spread within bacterial populations. The pCF10 plasmid system is the best characterised, and although other PRP systems are structurally similar, they lack exact functional homologues of pCF10-encoded genes. In this review, we provide an overview of the enterococcal PRP systems, incorporating functional details for the less-well-defined systems. We catalogue the virulence-associated elements of the PRPs that have been identified to date, and we argue that this reinforces the requirement for elucidation of the less studied systems.