Decolonization in Nursing Homes to Prevent Infection and Hospitalization.

BACKGROUND: Nursing home residents are at high risk for infection, hospitalization, and colonization with multidrug-resistant organisms. METHODS: We performed a cluster-randomized trial of universal decolonization as compared with routine-care bathing in nursing homes. The trial included an 18-month baseline period and an 18-month intervention period. Decolonization entailed the use of chlorhexidine for all routine bathing and showering and administration of nasal povidone-iodine twice daily for the first 5 days after admission and then twice daily for 5 days every other week. The primary outcome was transfer to a hospital due to infection. The secondary outcome was transfer to a hospital for any reason. An intention-to-treat (as-assigned) difference-in-differences analysis was performed for each outcome with the use of generalized linear mixed models to compare the intervention period with the baseline period across trial groups. RESULTS: Data were obtained from 28 nursing homes with a total of 28,956 residents. Among the transfers to a hospital in the routine-care group, 62.2% (the mean across facilities) were due to infection during the baseline period and 62.6% were due to infection during the intervention period (risk ratio, 1.00; 95% confidence interval [CI], 0.96 to 1.04). The corresponding values in the decolonization group were 62.9% and 52.2% (risk ratio, 0.83; 95% CI, 0.79 to 0.88), for a difference in risk ratio, as compared with routine care, of 16.6% (95% CI, 11.0 to 21.8; P<0.001). Among the discharges from the nursing home in the routine-care group, transfer to a hospital for any reason accounted for 36.6% during the baseline period and for 39.2% during the intervention period (risk ratio, 1.08; 95% CI, 1.04 to 1.12). The corresponding values in the decolonization group were 35.5% and 32.4% (risk ratio, 0.92; 95% CI, 0.88 to 0.96), for a difference in risk ratio, as compared with routine care, of 14.6% (95% CI, 9.7 to 19.2). The number needed to treat was 9.7 to prevent one infection-related hospitalization and 8.9 to prevent one hospitalization for any reason. CONCLUSIONS: In nursing homes, universal decolonization with chlorhexidine and nasal iodophor led to a significantly lower risk of transfer to a hospital due to infection than routine care. (Funded by the Agency for Healthcare Research and Quality; Protect ClinicalTrials.gov number, NCT03118232.).

Reducing Hospitalizations and Multidrug-Resistant Organisms via Regional Decolonization in Hospitals and Nursing Homes.

Importance: Infections due to multidrug-resistant organisms (MDROs) are associated with increased morbidity, mortality, length of hospitalization, and health care costs. Regional interventions may be advantageous in mitigating MDROs and associated infections. Objective: To evaluate whether implementation of a decolonization collaborative is associated with reduced regional MDRO prevalence, incident clinical cultures, infection-related hospitalizations, costs, and deaths. Design, Setting, and Participants: This quality improvement study was conducted from July 1, 2017, to July 31, 2019, across 35 health care facilities in Orange County, California. Exposures: Chlorhexidine bathing and nasal iodophor antisepsis for residents in long-term care and hospitalized patients in contact precautions (CP). Main Outcomes and Measures: Baseline and end of intervention MDRO point prevalence among participating facilities; incident MDRO (nonscreening) clinical cultures among participating and nonparticipating facilities; and infection-related hospitalizations and associated costs and deaths among residents in participating and nonparticipating nursing homes (NHs). Results: Thirty-five facilities (16 hospitals, 16 NHs, 3 long-term acute care hospitals [LTACHs]) adopted the intervention. Comparing decolonization with baseline periods among participating facilities, the mean (SD) MDRO prevalence decreased from 63.9% (12.2%) to 49.9% (11.3%) among NHs, from 80.0% (7.2%) to 53.3% (13.3%) among LTACHs (odds ratio [OR] for NHs and LTACHs, 0.48; 95% CI, 0.40-0.57), and from 64.1% (8.5%) to 55.4% (13.8%) (OR, 0.75; 95% CI, 0.60-0.93) among hospitalized patients in CP. When comparing decolonization with baseline among NHs, the mean (SD) monthly incident MDRO clinical cultures changed from 2.7 (1.9) to 1.7 (1.1) among participating NHs, from 1.7 (1.4) to 1.5 (1.1) among nonparticipating NHs (group × period interaction reduction, 30.4%; 95% CI, 16.4%-42.1%), from 25.5 (18.6) to 25.0 (15.9) among participating hospitals, from 12.5 (10.1) to 14.3 (10.2) among nonparticipating hospitals (group × period interaction reduction, 12.9%; 95% CI, 3.3%-21.5%), and from 14.8 (8.6) to 8.2 (6.1) among LTACHs (all facilities participating; 22.5% reduction; 95% CI, 4.4%-37.1%). For NHs, the rate of infection-related hospitalizations per 1000 resident-days changed from 2.31 during baseline to 1.94 during intervention among participating NHs, and from 1.90 to 2.03 among nonparticipating NHs (group × period interaction reduction, 26.7%; 95% CI, 19.0%-34.5%). Associated hospitalization costs per 1000 resident-days changed from $64 651 to $55 149 among participating NHs and from $55 151 to $59 327 among nonparticipating NHs (group × period interaction reduction, 26.8%; 95% CI, 26.7%-26.9%). Associated hospitalization deaths per 1000 resident-days changed from 0.29 to 0.25 among participating NHs and from 0.23 to 0.24 among nonparticipating NHs (group × period interaction reduction, 23.7%; 95% CI, 4.5%-43.0%). Conclusions and Relevance: A regional collaborative involving universal decolonization in long-term care facilities and targeted decolonization among hospital patients in CP was associated with lower MDRO carriage, infections, hospitalizations, costs, and deaths.

Early Infectious Diseases Specialty Intervention Is Associated With Shorter Hospital Stays and Lower Readmission Rates: A Retrospective Cohort Study.

Background: Intervention by infectious diseases (ID) physicians improves outcomes for inpatients in Medicare, but patients with other insurance types could fare differently. We assessed whether ID involvement leads to better outcomes among privately insured patients under age 65 years hospitalized with common infections. Methods: We performed a retrospective analysis of administrative claims data from community hospital and postdischarge ambulatory care. Patients were privately insured individuals less than 65 years old with an acute-care stay in 2014 for selected infections, classed as having early (by day 3) or late (after day 3) ID intervention, or none. Key outcomes were mortality, cost, length of the index stay, readmission rate, mortality, and total cost of care over the first 30 days after discharge. Results: Patients managed with early ID involvement had shorter length of stay, lower spending, and lower mortality in the index stay than those patients managed without ID involvement. Relative to late, early ID involvement was associated with shorter length of stay and lower cost. Individuals with early ID intervention during hospitalization had fewer readmissions and lower healthcare payments after discharge. Relative to late, those with early ID intervention experienced lower readmission, lower spending, and lower mortality. Conclusions: Among privately insured patients less than 65 years old, treated in a hospital, early intervention with an ID physician was associated with lower mortality rate and shorter length of stay. Patients who received early ID intervention during their hospital stay were less likely to be readmitted after discharge and had lower total healthcare spending.

The SHIELD Orange County Project: Multidrug-resistant Organism Prevalence in 21 Nursing Homes and Long-term Acute Care Facilities in Southern California.

BACKGROUND: Multidrug-resistant organisms (MDROs) spread between hospitals, nursing homes (NHs), and long-term acute care facilities (LTACs) via patient transfers. The Shared Healthcare Intervention to Eliminate Life-threatening Dissemination of MDROs in Orange County is a regional public health collaborative involving decolonization at 38 healthcare facilities selected based on their high degree of patient sharing. We report baseline MDRO prevalence in 21 NHs/LTACs. METHODS: A random sample of 50 adults for 21 NHs/LTACs (18 NHs, 3 LTACs) were screened for methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus spp. (VRE), extended-spectrum ß-lactamase-producing organisms (ESBL), and carbapenem-resistant Enterobacteriaceae (CRE) using nares, skin (axilla/groin), and peri-rectal swabs. Facility and resident characteristics associated with MDRO carriage were assessed using multivariable models clustering by person and facility. RESULTS: Prevalence of MDROs was 65% in NHs and 80% in LTACs. The most common MDROs in NHs were MRSA (42%) and ESBL (34%); in LTACs they were VRE (55%) and ESBL (38%). CRE prevalence was higher in facilities that manage ventilated LTAC patients and NH residents (8% vs <1%, P < .001). MDRO status was known for 18% of NH residents and 49% of LTAC patients. MDRO-colonized adults commonly harbored additional MDROs (54% MDRO+ NH residents and 62% MDRO+ LTACs patients). History of MRSA (odds ratio [OR] = 1.7; confidence interval [CI]: 1.2, 2.4; P = .004), VRE (OR = 2.1; CI: 1.2, 3.8; P = .01), ESBL (OR = 1.6; CI: 1.1, 2.3; P = .03), and diabetes (OR = 1.3; CI: 1.0, 1.7; P = .03) were associated with any MDRO carriage. CONCLUSIONS: The majority of NH residents and LTAC patients harbor MDROs. MDRO status is frequently unknown to the facility. The high MDRO prevalence highlights the need for prevention efforts in NHs/LTACs as part of regional efforts to control MDRO spread.

Essential Resources and Strategies for Antibiotic Stewardship Programs in the Acute Care Setting.

Background: Antibiotic stewardship programs improve clinical outcomes and patient safety and help combat antibiotic resistance. Specific guidance on resources needed to structure stewardship programs is lacking. This manuscript describes results of a survey of US stewardship programs and resultant recommendations regarding potential staffing structures in the acute care setting. Methods: A cross-sectional survey of members of 3 infectious diseases subspecialty societies actively involved in antibiotic stewardship was conducted. Survey responses were analyzed with descriptive statistics. Logistic regression models were used to investigate the relationship between stewardship program staffing levels and self-reported effectiveness and to determine which strategies mediate effectiveness. Results: Two-hundred forty-four respondents from a variety of acute care settings completed the survey. Prior authorization for select antibiotics, antibiotic reviews with prospective audit and feedback, and guideline development were common strategies. Eighty-five percent of surveyed programs demonstrated effectiveness in at least 1 outcome in the prior 2 years. Each 0.50 increase in pharmacist and physician full-time equivalent (FTE) support predicted a 1.48-fold increase in the odds of demonstrating effectiveness. The effect was mediated by the ability to perform prospective audit and feedback. Most programs noted significant barriers to success. Conclusions: Based on our survey's results, we propose an FTE-to-bed ratio that can be used as a starting point to guide discussions regarding necessary resources for antibiotic stewardship programs with executive leadership. Prospective audit and feedback should be the cornerstone of stewardship programs, and both physician leadership and pharmacists with expertise in stewardship are crucial for success.

Human Homolog of Drosophila Ariadne (HHARI) is a marker of cellular proliferation associated with nuclear bodies.

HHARI (also known as ARIH1) is an ubiquitin-protein ligase and is the cognate of the E2, UbcH7 (UBE2L3). To establish a functional role for HHARI in cellular proliferation processes, we performed a reverse genetics screen that identified n=86/522 (16.5%) ubiquitin conjugation components that have a statistically significant effect on cell proliferation, which included HHARI as a strong hit. We then produced and validated a panel of specific antibodies that establish HHARI as both a nuclear and cytoplasmic protein that is expressed in all cell types studied. HHARI was expressed at higher levels in nuclei, and co-localized with nuclear bodies including Cajal bodies (p80 coilin, NOPP140), PML and SC35 bodies. We confirmed reduced cellular proliferation after ARIH1 knockdown with individual siRNA duplexes, in addition to significantly increased levels of apoptosis, an increased proportion of cells in G2 phase of the cell cycle, and significant reductions in total cellular RNA levels. In head and neck squamous cell carcinoma biopsies, there are higher levels of HHARI expression associated with increased levels of proliferation, compared to healthy control tissues. We demonstrate that HHARI is associated with cellular proliferation, which may be mediated through its interaction with UbcH7 and modification of proteins in nuclear bodies.

Identifying veterinary surgeons' barriers to, and potential solutions for, improving antimicrobial stewardship among sheep farmers in Northern Ireland.

Background: In order to improve antimicrobial stewardship (AMS), including changes in antimicrobial prescribing and use, an enhanced understanding is needed of the barriers that veterinary surgeons (vets) encounter to institute such change. Methods: A qualitative approach, using grounded theory, was followed. Interviews and discussion groups, with vets and farm industry stakeholders in Northern Ireland (NI), were undertaken to identify and explore attitudes and behaviours surrounding AMS, with a particular emphasis on the barriers vets encountered and the context within which they were working. Results: Seven inter-related themes associated with improving AMS among their sheep farming clients were identified. The first six addressed barriers were working under commercial and practical constraints, farmer behaviour, multiple medicine sources, poor prescribing practice, a perceived lack of incentive or facilitation to improve AMS and a perceived lack of action by regulators to challenge poor AMS. The seventh theme revealed suggestions vets considered that may improve AMS in NI, including greater state intervention in recording and regulating medicine sales. Conclusions: Improving AMS will require vets and their client farmers to change behaviour. This will involve concerted effort over an extended period of time to enact and embed change. Veterinary surgeons believe that further action by the industry and state to develop centralised antimicrobial sales recording and by the state to enforce prescribing regulations will aid their efforts. However, critical to achieving this is the development of a sustainable and funded mechanism to create more meaningful farmer-vet consultation around flock health prior to every prescription to improve AMS and sheep welfare.

Antibiotic use in the Northern Irish sheep flock: What lessons can be learnt from medicine records and farmer attitudes to improve stewardship of these essential medicines?

Stewardship of antibiotics used in livestock production has come under increasing scrutiny, from both the animal welfare point of view and due to concerns that antibiotic use in livestock may pose a risk to human health through selection pressure to drive development of antibiotic resistant strains of bacteria. Despite this concern, however, antibiotic consumption in the sheep sector is currently poorly described. This study determines the range and quantities of antibiotics used in the Northern Irish (NI) sheep flock as well as exploring drivers for their use. A mixed-methods approach was utilised, with an anonymous online scoping survey, analysis of the medicine records from 52 NI sheep farms and semi-structured interviews undertaken with 27 farmers. Eighteen farmers contributed both records and participated in interviews. Veterinary medicine records were derived from two sources: on-farm medicine books (seven) or veterinary practice sales data (51). As six of these farmers provided information from both sources a total of 52 unique farms participated. Overall, antibiotic use in sheep on the 52 farms sampled was low, with a median value of 11.35 mgPCU-1 (mean 13.63 mgPCU-1, sd 10.7; range 0-45.29 mgPCU-1), with all farms below 50 mgkg-1. Critically important antibiotics accounted for 0.21% of all antibiotics purchased. Lameness was the main driver of antibiotic use identified by this study. Others included a range of prophylactic treatments such as oral antibiotics to prevent watery mouth, injectable antibiotics to prevent abortion and following assisted lambing. Farmers acknowledged some of these uses had become habitual over time. The veterinary medicine sales records demonstrated significant sales of antibiotics not authorised for use in sheep, on an ongoing, rather than case-by-case, basis. Farmers were positive about their veterinarian's ability and knowledge to improve flock welfare and productivity, but were unwilling to pay for this advice. However, veterinarians may have facilitated weak medicine stewardship through a failure to adequately challenge farmers seeking antibiotics. Farmers did not maintain accurate or up-to-date on farm medicine or production records in the majority of cases. Despite this lack of on-farm recording, veterinary sales records can be studied in consultation with farmers to provide veterinarians with a farm-specific insight into antibiotic use and related attitudes and behaviours. Farmers and veterinarians can then identify areas and behaviours to target collaboratively, improving antibiotic and wider medicine stewardship, whilst simultaneously improving flock health and productivity.

Assessing past versus present severe acute respiratory coronavirus virus 2 (SARS-CoV-2) infection: A survey of criteria for discontinuing precautions in asymptomatic patients testing positive on admission.

Infection prevention program leaders report frequent use of criteria to distinguish recently recovered coronavirus disease 2019 (COVID-19) cases from actively infectious cases when incidentally positive asymptomatic patients were identified on routine severe acute respiratory coronavirus virus 2 (SARS-CoV-2) polymerase chain reaction (PCR) testing. Guidance on appropriate interpretation of high-sensitivity molecular tests can prevent harm from unnecessary precautions that delay admission and impede medical care.

Molecular chaperone-mediated rescue of mitophagy by a Parkin RING1 domain mutant.

Mitochondrial dysfunction is characteristic of many neurodegenerative diseases. The Parkinson's disease-associated ubiquitin-protein ligase, Parkin, is important in the elimination of damaged mitochondria by autophagy (mitophagy) in a multistep process. Here, we show that a Parkin RING domain mutant (C289G) fails to redistribute to damaged mitochondria and cannot induce mitophagy after treatment with the mitochondrial uncoupler carbonyl cyanide m-methylhydrazone, because of protein misfolding and aggregation. Parkin(C289G) aggregation and inclusion formation were suppressed by the neuronal DnaJ/Hsp40 chaperone HSJ1a(DNAJB2a). Importantly, HSJ1a and DNAJB6 also restored mitophagy by promoting the relocation of Parkin(C289G) and the autophagy marker LC3 to depolarized mitochondria. The rescue of Parkin activity and suppression of aggregation were J domain dependent for HSJ1a, suggesting the involvement of Hsp70 in these processes, but were not dependent on the HSJ1a ubiquitin interaction motif. HSJ1a expression did not enhance mitophagy mediated by wild-type Parkin. These data show the potential of molecular chaperones to mediate the functional recovery of Parkin misfolding mutants and to combat deficits associated with Parkin aggregation in Parkinson's disease.

Improving analgesia provision for sheep: An analysis of farm medicine records and attitudes towards pain relief on sheep farms in Northern Ireland.

Background: Management of pain is critical to improve the welfare of farmed livestock and meet consumer expectations. There is limited published information about the use of analgesic drugs in the sheep sector. Methods: A mixed-method approach was followed. The range of analgesic drugs used on 52 Northern Irish sheep farms was determined through analysis of medicine purchase records. Through interview and discussion groups, with both farmer and veterinarian participants, attitudes towards the use and adoption of such medicines were explored. Results: The use of non-steroidal anti-inflammatory drugs (NSAIDs) was widespread and highly variable. One-third of farmers in the sample did not purchase any NSAID. Meloxicam was the most commonly purchased NSAID by mass (72%) and standardised dose (73%). During interviews and discussions, farmers outlined the benefits they saw in using NSAIDs and how veterinarians influenced their uptake of these medicines. Use of corticosteroid was evidenced on 50% of the farms that supplied medicine records for analysis. Conclusions: Veterinarians can influence farmers to adopt NSAIDs for the provision of analgesia in their sheep and farmers observed the benefits they delivered. However, many farmers are still to be reached with this message, perhaps due to being largely self-sufficient and rarely engaging with veterinarians. Veterinarians have the opportunity to challenge farmers about the provision of analgesia, especially when farmers seek antibiotics for painful conditions such as lameness. Currently, the lack of an authorised product in the UK, with associated treatment guidance and industry promotion, may limit veterinarians' confidence in prescribing drugs for pain control in sheep.

Regional skeletal muscle remodeling and mitochondrial dysfunction in right ventricular heart failure.

Exercise intolerance is a cardinal symptom of right ventricular heart failure (RV HF) and skeletal muscle adaptations play a role in this limitation. We determined regional remodeling of muscle structure and mitochondrial function in a rat model of RV HF induced by monocrotaline injection (MCT; 60 mg·kg(-1); n = 11). Serial sections of the plantaris were stained for fiber type, succinate dehydrogenase (SDH) activity and capillaries. Mitochondrial function was assessed in permeabilized fibers using respirometry, and isolated complex activity by blue native gel electrophoresis (BN PAGE). All measurements were compared with saline-injected control animals (CON; n = 12). Overall fiber cross-sectional area was smaller in MCT than CON: 1,843 ± 114 vs. 2,322 ± 120 µm(2) (P = 0.009). Capillary-to-fiber ratio was lower in MCT in the oxidative plantaris region (1.65 ± 0.09 vs. 1.93 ± 0.07; P = 0.03), but not in the glycolytic region. SDH activity (P = 0.048) and maximal respiratory rate (P = 0.012) were each ∼15% lower in all fibers in MCT. ADP sensitivity was reduced in both skeletal muscle regions in MCT (P = 0.032), but normalized by rotenone. A 20% lower complex I/IV activity in MCT was confirmed by BN PAGE. MCT-treatment was associated with lower mitochondrial volume density (lower SDH activity), quality (lower complex I activity), and fewer capillaries per fiber area in oxidative skeletal muscle. These features are consistent with structural and functional remodeling of the determinants of oxygen supply potential and utilization that may contribute to exercise intolerance and reduced quality of life in patients with RV HF.

Involvement of X-box binding protein 1 and reactive oxygen species pathways in the pathogenesis of tumour necrosis factor receptor-associated periodic syndrome.

OBJECTIVES: To investigate convergence of endoplasmic reticulum stress pathways and enhanced reactive oxygen species (ROS) production, due to intracellular retention of mutant tumour necrosis factor receptor 1 (TNFR1), as a disease mechanism in TNFR-associated periodic syndrome (TRAPS). METHODS: Peripheral blood mononuclear cells from patients with TRAPS (n=16) and healthy controls (HC) (n=22) were studied alongside HEK293T cells expressing wild type-TNFR1 or TRAPS-associated mutations. Unfolded protein response (UPR)-associated proteins (protein kinase-like endoplasmic reticulum kinase, PERK), phosphorylated-PERK (p-PERK), phosphorylated inositol-requiring enzyme 1α (p-IRE1α) and spliced X-box binding protein 1 (sXBP1)) were measured by flow cytometry. XBP1 splicing and UPR-associated transcript expression were assessed by reverse transcription PCR/quantitative real-time PCR. ROS levels were measured using CM-H(2)DCFDA and MitoSOX Red in patients' monocytes or HEK293T cells by flow cytometry. RESULTS: Mutant TNFR1-expressing HEK293T cells had increased TNFR1 expression associated with intracellular aggregation. TRAPS patients had increased sXBP1 transcripts (p<0.01) compared with HC. Raised p-PERK protein was seen, indicative of an UPR, but other UPR-associated transcripts were normal. Increased ROS levels were observed in TRAPS monocytes compared with HCs (p<0.02); these increased further upon IL-6 stimulation (p<0.01). Lipopolysaccharide-stimulated peripheral blood mononuclear cells of patients with TRAPS, but not HCs, demonstrated increased sXBP1 levels (p<0.01), which were reduced by antioxidant treatment (p<0.05). CONCLUSIONS: Patients with TRAPS have evidence of increased sXBP1 and PERK expression but without other signs of classical UPR, and also with high ROS generation that may contribute to the pro-inflammatory state associated with TRAPS. The authors propose a non-traditional XBP1 pathway with enhanced sXBP1 as a novel disease-contributing mechanism in TRAPS.

Drivers and hazards of consumption of unpasteurised bovine milk and milk products in high-income countries.

Introduction: The consumption of dairy products contributes to health, nutrition, and livelihoods globally. However, dairy products do not come without microbiological food safety risks for consumers. Despite this risk, common hygiene measures in high-income countries, particularly pasteurisation, ensures that milk is safe, and is indeed frequently mandated by law. Nevertheless, over the past two decades, there has been a global increase in the number of consumers in high-income developed countries actively seeking out unpasteurised milk in liquid and product forms for perceived nutritional and health benefits, and improved taste. The often-anecdotal claims upon which consumers make such choices are not all supported by scientific evidence; however, some recent research studies have investigated (and in some cases demonstrated) the positive impact of unpasteurised milk consumption on the prevalence of asthma, atopy, rectal cancer and respiratory illness. Methods: To investigate the significance of unpasteurised milk and milk product consumption for human health in high-income countries, outbreak data between the years 2000 and 2018 were obtained for the United States of America, Canada, the European Union, the United Kingdom, Japan, New Zealand and Australia, which were then categorized into three World Health Organisation subregions: AMR A, EUR A and WPR A. Outbreak dynamic variables such as pathogens, the place of consumption, numbers of outbreaks and deaths per million capita, the average number of cases per outbreak and regulations were described and analysed using R Studio. To provide an overview of unpasteurised milk-related disease outbreaks, a rapid evidence review was also undertaken to establish an overview of what is known in the current literature about hazards and drivers of consumption. Results: Foodborne outbreaks associated with unpasteurised dairy consumption have risen in high-income countries over the period 2000 to 2018, with Campylobacter spp. being the most common aetiological agent responsible, followed by Escherichia coli and Salmonella spp. The most common places of consumption are on farms or in households, indicating individuals choose to drink unpasteurised milk, rather than a widespread distribution of the product, for example, at social events and in schools. Further study is needed to better understand contributing factors, such as cultural differences in the consumption of dairy products. Conclusion: There are several observable health benefits linked to consuming raw milk, but outbreaks associated with unpasteurised milk and milk products are on the rise. It cannot be definitively concluded whether the benefits outweigh the risks, and ultimately the decision lies with the individual consumer. Nevertheless, many countries have regulations in place to protect consumer health, acknowledging the definite risks to human health that unpasteurised dairy foods may pose, particularly from microbial hazards.

Sheep scab in Northern Ireland: Its distribution, costs and farmer knowledge about prevention and control.

Sheep scab (psoroptic mange), which is endemic in the United Kingdom (UK) flock, has a significant, negative impact on sheep welfare. Nothing has previously been published about the distribution of sheep scab in Northern Ireland (NI), nor about Northern Irish farmers' knowledge and behaviours relating to the disease, its treatment, prevention and control. Between March and June 2021 an online questionnaire on the disease was completed by sheep farmers in NI. Forty-four respondents out of a total of 122 valid returns (36%) indicated that they had at least one outbreak of sheep scab in their flock within the previous five years. These flocks were spread throughout NI and included flocks grazing on common land. Farmers reporting sheep scab in their flock considered movements of sheep between flocks to be the main cause of flock infestation. Respondents demonstrated knowledge gaps in relation to the parasite biology, disease transmission, prevention and treatment options, as well as a lack of awareness of some of the relevant industry guidelines. We highlight that some farmers rely on clinical signs alone to rule out the possibility that newly purchased sheep are infested with sheep scab before mixing them with their flock. This activity poses a high risk for the introduction of sheep scab into previously uninfested flocks. The inadequacy of some farmers' quarantine rules, or their inability to follow them, was also reported by farmers as being the cause of their flock infestation. Sheep scab outbreaks were shown to result in significant financial cost, with some farmers reporting their most recent outbreak had cost over £2500 ($3329). The paper also highlights that in addition to the animal health and welfare impact and financial cost, sheep scab was reported to have a social cost: 94 respondents (79%) agreed that a sheep scab outbreak caused emotional stress to affected farmers. These findings have provided evidence of the widespread nature of sheep scab in the NI flock, and of the knowledge gaps and behaviours which need to be addressed to improve sheep scab control. This will require a combination of focused research, knowledge exchange between farmers, advisors, policy makers and regulators, and co-developed disease control plans at a flock and national level.

A Nonrestrictive Approach to Fluoroquinolone Stewardship at Two Community Hospitals.

Background: Fluoroquinolones are one of the most prescribed antimicrobials in the United States and have been increasingly used in inpatient and outpatient settings to treat various infectious diseases syndromes. Due to the unwanted collateral effects on antibiotic resistance, poor susceptibility rates among Gram-negative pathogens, and adverse effects, fluoroquinolones are often targeted by hospital antimicrobial stewardship programs to prevent overutilization. This study describes the association of nonrestrictive antimicrobial stewardship interventions at 2 nonacademic community hospitals on levofloxacin utilization, prescribing patterns on alternative antibiotics, and Pseudomonas aeruginosa nonsusceptibility rates to levofloxacin. Methods: Nonrestrictive antimicrobial stewardship interventions included monitoring and reporting of fluoroquinolone susceptibility trends to physician groups, performing medication use evaluations of levofloxacin accompanied with prescriber detailing, daily prospective audit and feedback, implementation of beta-lactam-based institutional guidelines for empiric therapy in various infectious disease syndromes, review and adjustment of electronic medical record order sets containing fluoroquinolones, and intensive prescriber education. No preauthorization of levofloxacin was used during this study period. Antibiotic utilization data were collected for the time periods of August 2015 through January 2021. Correlation between levofloxacin and other broad-spectrum antibiotc use was investigated as well as the impact on Pseudomonas aeruginosa levofloxacin nonsusceptibility rates. Results: Both hospitals showed an overall downward trend in the prescribing of levofloxacin during the time period of August 2015 to January 2021. There was a significant negative correlation between monthly ceftriaxone and levofloxacin days of therapy for both hospitals (P < .0001). There was a positive correlation between levofloxacin days of therapy and P aeruginosa nonsusceptibility (P < .02 at both hospitals). Conclusions: Our results demonstrate that a nonrestrictive approach to fluoroquinolone stewardship interventions had a significant impact on reducing levofloxacin utilization, increasing ceftriaxone utilization, and improving P aeruginosa levofloxacin susceptibility.

Regulation of canonical Wnt signalling by the ciliopathy protein MKS1 and the E2 ubiquitin-conjugating enzyme UBE2E1.

Primary ciliary defects cause a group of developmental conditions known as ciliopathies. Here, we provide mechanistic insight into ciliary ubiquitin processing in cells and for mouse model lacking the ciliary protein Mks1. In vivo loss of Mks1 sensitises cells to proteasomal disruption, leading to abnormal accumulation of ubiquitinated proteins. We identified UBE2E1, an E2 ubiquitin-conjugating enzyme that polyubiquitinates ß-catenin, and RNF34, an E3 ligase, as novel interactants of MKS1. UBE2E1 and MKS1 colocalised, and loss of UBE2E1 recapitulates the ciliary and Wnt signalling phenotypes observed during loss of MKS1. Levels of UBE2E1 and MKS1 are co-dependent and UBE2E1 mediates both regulatory and degradative ubiquitination of MKS1. We demonstrate that processing of phosphorylated ß-catenin occurs at the ciliary base through the functional interaction between UBE2E1 and MKS1. These observations suggest that correct ß-catenin levels are tightly regulated at the primary cilium by a ciliary-specific E2 (UBE2E1) and a regulatory substrate-adaptor (MKS1).

A simple cell based assay to measure Parkin activity.

Parkin is an ubiquitin-protein ligase mutated in Autosomal Recessive - Juvenile Parkinsonism. Here, we describe a cell-based assay to measure Parkin's ubiquitin-protein ligase activity. It relies on the ability of Parkin to recognise depolarised mitochondria and exploits a cell line where Parkin expression is inducible. In these cells, Parkin expression promotes mitophagy and accelerates cell death in response to mitochondrial depolarisers. Time-lapse imaging confirmed cell death and revealed increased perinuclear mitochondrial clustering following induction of Parkin expression in cells exposed to carbonyl cyanide m-chlorophenylhydrazone. Similar effects were not observed with α-synuclein or DJ-1, other proteins associated with the development of Parkinson's disease, confirming the specificity of the assay. We have used this assay to demonstrate that ligase-defective Parkin mutants are inactive, and cellular proteasomal activity (using the proteasomal inhibitors MG132, clasto-lactacystin ß-lactone and epoxomicin) is essential for the Parkin mediated effect. As the assay is suitable for high-throughput screening, it has the potential to identify novel proteostasis compounds that stimulate the activity of Parkin mutants for therapeutic purposes, to identify modulators of kinase activities that impact on Parkin function, and to act as a functional read-out in reverse genetics screens aimed at identifying modifiers of Parkin function during mitophagy.

Why isn't the transition period getting the attention it deserves? Farm advisors' opinions and experiences of managing dairy cow health in the transition period.

During the transition period three weeks before and after calving the dairy cow is at greater risk of developing disease, to the detriment of welfare and production. An understanding of the reasons why and how farmers and their advisors engage in efforts to control metabolic disease during the transition period is required if these diseases are to be more successfully controlled. The study reported here, based on interview research, investigates the opinions and behaviours of farm advisors on transition cow management and nutrition, their experiences of working with their respective farm clients, and interactions with other farm advisors to help manage transition cow health and productivity. Semi-structured interviews were conducted with 12 veterinary advisors and 12 non-veterinary advisors (nutritionists, feed company representatives and independent consultants) in England. A key theme emerging from this qualitative data was a perceived lack of focussed transition management advice provided by advisors. Reasons for suboptimal or lack of appropriate advice included: time pressures for advisors to visit as many farms as possible; avoiding the investigation of areas of potential improvement, for fear of not meeting transition health and performance targets; financial disincentives for nutritionists, as the sales commission attributed to transition cow feeding was small relative to the main milking herd; and a lack of confidence in the subject. Other aspects included the responsibility of providing transition advice which was perceived to be high-risk, a lack of cooperation between veterinarians and nutritionists, and the perceived varying competencies of nutritionists. The findings demonstrate the importance of the varied influences of 'people factors' on transition cow health such as the nature of the advisor-farmer relationship, advisor-farmer communication and herd-level advisor collaboration on transition cow health and management.

Dairy cow health and management in the transition period: The need to understand the human dimension.

During the transition from the dry period to lactation the dairy cow undergoes a period of physiological, metabolic and immunological change, and is at greater risk of developing disease, to the detriment of health, welfare and production. Many studies have been undertaken to determine appropriate management strategies to improve health and welfare during the transition period, however the incidence of disease, particularly metabolic disease, in this period remains high. To date, a lack of research attention has been paid to the social factors which may affect the management of transition dairy cows. An understanding of farmer and advisor attitudes and behaviour, and the challenges they face in managing transition cows, may help to direct farmers towards more effective disease prevention and control. It is also possible that transition cow morbidity may be due to complex interactions that are difficult to manage, despite efforts to implement best practice. This review paper provides a brief overview of some of the management factors that may influence herd health during the transition period. It then investigates how social influences may relate to the uptake of transition management practices by exploring the use of qualitative interviews investigating farmer and stakeholder attitudes and behaviour in relation to cattle health and welfare, before focussing more specifically on farmer behaviour. Additionally, this paper explores farm advisor behaviour, and how that has been shown to influence farmer adherence to advice, which has particular relevance to transition cow management. It then suggests potential research strategies to investigate the human influences affecting the scale of the problem that may provide solutions to tackle the challenge of improving dairy cow health and welfare.