US3 Kinase-Mediated Phosphorylation of Tegument Protein VP8 Plays a Critical Role in the Cellular Localization of VP8 and Its Effect on the Lipid Metabolism of Bovine Herpesvirus 1-Infected Cells.

Bovine herpesvirus 1 (BoHV-1) infects bovine species, causing respiratory infections, genital disorders and abortions. VP8 is the most abundant tegument protein of BoHV-1 and is critical for virus replication in cattle. In this study, the cellular transport of VP8 in BoHV-1-infected cells and its ability to alter the cellular lipid metabolism were investigated. A viral kinase, US3, was found to be involved in regulating these processes. In the early stages of infection VP8 was localized in the nucleus. Subsequently, presumably after completion of its role in the nucleus, VP8 was translocated to the cytoplasm. When US3 was deleted or the essential US3 phosphorylation site of VP8 was mutated in BoHV-1, the majority of VP8 was localized in the nuclei of infected cells. This suggests that phosphorylation by US3 may be critical for cytoplasmic localization of VP8. Eventually, the cytoplasmic VP8 was accumulated in the cis-Golgi apparatus but not in the trans-Golgi network, implying that VP8 was not involved in virion transport toward and budding from the cell membrane. VP8 caused lipid droplet (LD) formation in the nuclei of transfected cells and increased cellular cholesterol levels. Lipid droplets were not found in the nuclei of BoHV-1-infected cells when VP8 was cytoplasmic in the presence of US3. However, when US3 was deleted or phosphorylation residues in VP8 were mutated, nuclear VP8 and LDs appeared in BoHV-1-infected cells. The total cholesterol level was increased in BoHV-1-infected cells but not in ΔUL47-BoHV-1-infected cells, further supporting a role for VP8 in altering the cellular lipid metabolism during infection.IMPORTANCE Nuclear localization signals (NLSs) and nuclear export signals (NESs) are important elements directing VP8 to the desired locations in the BoHV-1-infected cell. In this study, a critical regulator that switches the nuclear and cytoplasmic localization of VP8 in BoHV-1-infected cells was identified. BoHV-1 used viral kinase US3 to regulate the cellular localization of VP8. Early during BoHV-1 infection VP8 was localized in the nucleus, where it performs various functions; once US3 was expressed, phosphorylated VP8 was cytoplasmic and ultimately accumulated in the cis-Golgi apparatus, presumably to be incorporated into virions. The Golgi localization of VP8 was only observed in virus-infected cells and not in US3-cotransfected cells, suggesting that this is mediated by other viral factors. Interestingly, VP8 was shown to cause increased cholesterol levels, which is a novel function for VP8 and a potential strategy to supply lipid for viral replication.

Escherichia coli and Klebsiella pneumoniae producing CTX-M cephalosporinase from swine finishing barns and their association with antimicrobial use.

We report the recovery of Escherichia coli or Klebsiella pneumoniae containing the extended-spectrum ß-lactamase gene bla(CTX-M) from 24 of 1,495 (1.6%) swine fecal samples in 8 of 50 (16%) finishing barns located in 5 U.S. states. We did not detect an association between antimicrobial use and recovery of bla(CTX-M).

Evaluation of Ultraviolet Type C Radiation in Inactivating Relevant Veterinary Viruses on Experimentally Contaminated Surfaces.

Many swine farms employ UVC treatment in employees' personal belongings and small tools entering farms as part of the biosecurity protocol to decrease the risk of pathogen introduction into the operation. However, the UVC efficacy in some veterinary viruses is not fully evaluated. This study evaluated the efficacy of ultraviolet type C (UVC) radiation in inactivating seven relevant veterinary viruses: Swine Poxvirus (SwPV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Porcine Epidemic Diarrhea Virus (PEDV), Swine Influenza Virus (SIV), Bovine Viral Diarrhea Virus (BVDV), Porcine Parvovirus (PPV), and Senecavirus A (SVA). The experimentally contaminated materials included polystyrene and filter paper. The samples were exposed to UVC for 5 min (total dose of 360 mJ/cm2). The UVC treatment caused a decrease over 4 log10 in SwPV titer on the polystyrene surface, whereas it consistently reduced about 5 log10 in PPV and SVA samples. No viable virus was recovered from PRRSV, PEDV, SIV, and BVDV samples. In filter paper, conversely, the efficacy was reduced. This study provides essential information on the inactivation effectiveness of a specific dose of UVC on important veterinary viruses, further supporting the rational application and strategic guidance for UVC radiation use to disinfect materials.

Employee absenteeism based on occupational health visits in an urban tertiary care Canadian hospital.

(1) To merge Occupational Health (OH) and Human Resources (HR) administrative data to describe reasons for absenteeism among hospital employees and (2) to consider the advantages and disadvantages of using these combined data for surveillance of health care workers. This study utilized a retrospective cohort design, involving a record linkage of two administrative databases at a Canadian general hospital: OH and HR. Data were included for the period of June 1, 2004, to May 31, 2005. Data linkage was performed using sex, postal code, and date of birth. The most common self-reported reasons for absence were respiratory illness (31%), gastrointestinal illness (17%), and musculoskeletal injuries/disabilities (15%). Employees working in the Department of General Medicine experienced the highest number of times absent--1.9 per 1,000 work hours. The department with the highest percentage of staff not reporting to OH was General Medicine (43%). This research highlights the issue of absenteeism among health care workers and the need to improve reporting of illness and injury to OH for surveillance efficacy. Further, a public health surveillance system that monitors OH visits among health care workers can facilitate public health practice.

Influenza vaccination and intention to receive the pandemic H1N1 influenza vaccine among healthcare workers of British Columbia, Canada: a cross-sectional study.

OBJECTIVE: To assess healthcare workers' attitudes and concerns regarding seasonal and pandemic influenza vaccines in order to improve vaccination campaign communications. DESIGN: Cross-sectional survey. SETTING: All 6 health authorities in British Columbia, Canada. METHODS: An anonymous, self-administered online survey was conducted from August 30 through September 30, 2009. Question topics included demographic characteristics, factors influencing acceptance of seasonal influenza vaccine, factors influencing intentions to accept pandemic H1N1 influenza vaccine, and knowledge and concerns regarding the effect of the influenza pandemic. PARTICIPANTS: All 96,217 British Columbia healthcare workers were eligible to participate. RESULTS: A volunteer sample of 4,046 healthcare workers returned the survey; 3,563 (88%) were women, 58% were under 50 years old (mean age + or - standard deviation, 45.3 + or - 10.9 years), 3,152 of 4,023 (79%) had 5 or more years of experience in their profession, 1,853 of 4,023 (46%) were nurses, and 2,833 (70%) had been vaccinated against seasonal influenza the previous year. Two thousand eight hundred (69%) respondents reported intending to receive the pandemic H1N1 vaccine. The most important predictor of this intention was having received the seasonal vaccine the previous year (odds ratio [OR], 6.25 [95% confidence interval {CI}, 5.39-7.26]). Worry about making loved ones ill was the only attitude associated with intention to receive the pandemic H1N1 vaccine (adjusted OR, 1.38 [95% CI, 1.27-1.50]). Concerns with vaccine safety (adjusted OR, 0.31 [95% CI, 0.25-0.39]) and belief "that H1N1 is not severe enough" (adjusted OR, 0.29 [95% CI, 0.26-0.32]) were independently associated with the intention to reject the pandemic H1N1 vaccine. CONCLUSIONS: Vaccination campaigns for pandemic H1N1 vaccine should use messages that emphasize the risk of illness among younger people and the opportunity to protect loved ones by getting the vaccine and should address concerns about the safety and effectiveness of the vaccine.