The effect of topical ophthalmic 1% atropine on heart rate and rhythm in normal dogs.

OBJECTIVE: To determine if topically administered ophthalmic atropine affects heart rate or rhythm in clinically normal dogs. METHODS: Two groups of 15 healthy dogs were evaluated, one consisting of dogs weighing <15 kg, the other consisting of dogs weighing >15 kg. Each dog was suited with a Holter monitor. At start time 0, dogs received one drop of ophthalmic 1% atropine solution, or one drop of sterile saline solution as a control in each eye, via random assignment. Each dog served as their own control. This procedure was repeated two more times, at 6-h intervals, for a total of three treatments over a 12-h period. Holter monitors recorded heart rate and rhythm for 24 h. Statistical analysis was performed to compare values between the groups. Dose dependent changes in cardiac parameters were evaluated. RESULTS: The mean heart rate and average minimum heart rate was significantly higher during the treatment period compared to the control period (8% and 13%, respectively). The mean number of hours with a heart rate <50 bpm decreased by 47% in the treatment vs. the control period. The mean maximum heart rate and number of h with a heart rate > 180 bpm did not differ significantly between the two groups. There was no evidence of dose dependence on heart rate when comparing small and large dogs. No significant differences in heart rhythm were noted between groups for measurable parameters. CONCLUSIONS: Topically administered atropine causes a small but significant increase in heart rate in healthy dogs.

Sexual behaviors and other risk factors for oral human papillomavirus infections in young women.

BACKGROUND: Oral human papillomavirus (HPV) is associated with a rising incidence of certain head and neck cancers, and oral sex has been associated with oral HPV. This study sought to identify more specific patterns of oral sexual activity, including self-inoculation, that are associated with oral HPV infections in young women. METHODS: A total of 1010 women attending a large university completed a computer-based questionnaire and provided oral specimens that were tested for any oral HPV using a Linear Array assay that detects any HPV as well as 37 HPV genotypes. Twenty-seven women provided additional samples up to 12 months after enrollment. Bivariable and multivariable analyses were conducted to identify oral sexual patterns and other risk factors associated with prevalent oral HPV. RESULTS: Nineteen women had prevalent oral HPV (1.9%), with 10 women (1%) having a type-specific infection. Oral HPV was significantly associated with lifetime coital sex partnership numbers (P = 0.03), lifetime and yearly oral sex partnership numbers (P < 0.01), and hand and/or sex toy transfer from genitals to mouth (P < 0.001). Oral HPV was also associated with greater use of alcohol, cigarettes, marijuana, and sharing of smoking devices, lipstick, or toothbrushes (P < 0.05 for each), with an apparent dose-response for alcohol use and smoking behavior, stratified by number of sexual partners. Of 7 women with prevalent HPV who provided follow-up samples, none had evidence of a persistent type-specific infection. CONCLUSIONS: These data provide additional evidence of transmission of oral HPV from oral sexual activity and also suggest possible transmission from self-inoculation or sharing of oral products.

New disease records for hatchery-reared sturgeon. I. Expansion of frog virus 3 host range into Scaphirhynchus albus.

In 2009, juvenile pallid sturgeon Scaphirhynchus albus, reared at the Blind Pony State Fish Hatchery (Missouri, USA) to replenish dwindling wild stocks, experienced mass mortality. Histological examination revealed extensive necrosis of the haematopoietic tissues, and a virus was isolated from affected organs in cell culture and then observed by electron microscopy. Experimental infection studies revealed that the virus is highly pathogenic to juvenile pallid sturgeon, one of several species of sturgeon currently listed as Endangered. The DNA sequence of the full length major capsid protein gene of the virus was identical to that of the species Frog virus 3 (FV3), the type species for the genus Ranavirus, originally isolated from northern leopard frog Lithobates pipiens. Although FV3 infections and epizootics in amphibians and reptiles are well documented, there is only 1 prior report of a natural infection of FV3 in fish. Our results illustrate the broad potential host range for FV3, with the known potential to cause significant mortality in poikilothermic vertebrates across 3 taxonomic classes including bony fishes, anuran and caudate amphibians, and squamate and testudine reptiles.

Serologic evidence of avian influenza virus infections among Nigerian agricultural workers.

Nigeria has had multiple incursions of highly pathogenic avian influenza A (HPAI) H5N1 virus into its poultry population since 2006. This study aimed to determine if Nigerians exposed to poultry had evidence of avian influenza virus transmission to man. Between 2008 and 2010, 316 adult farmers and open market workers and 54 age-group matched, non-animal exposed controls were enrolled in a prospective, population-based study of zoonotic influenza transmission in four towns in southeastern Nigeria. Questionnaire data and sera obtained at the time of enrollment were examined for evidence of previous infection with 10 avian influenza virus strains. Serologic studies on sera collected at the time of enrollment showed modest evidence of previous infection with three avian-origin influenza viruses (H5N1, H5N2, and H11N1) and one avian-like H9N2 influenza virus, with eight (2.4%) of animal-exposed subjects and two (3.7%) unexposed subjects having elevated microneutralization assay antibody titer levels (ranging from 1:10 to 1:80). Statistical analyses did not identify specific risk factors associated with the elevated antibody titers observed for these zoonotic influenza viruses. These data suggested only occasional virus transmission to humans in areas thought to have been enzootic for avian influenza virus. Prospective data from this cohort will help the authors to better understand the occurrence of zoonotic infections due to avian influenza viruses in Nigeria.

Influenza A(H1N1)pdm09 virus among healthy show pigs, United States.

Within 5 months after the earliest detection of human influenza A(H1N1)pdm09 virus, we found molecular and culture evidence of the virus in healthy US show pigs. The mixing of humans and pigs at swine shows possibly could further the geographic and cross-species spread of influenza A viruses.

Evidence for subclinical avian influenza virus infections among rural Thai villagers.

BACKGROUND: Regions of Thailand reported sporadic outbreaks of A/H5N1 highly pathogenic avian influenza (HPAI) among poultry between 2004 and 2008. Kamphaeng Phet Province, in north-central Thailand had over 50 HPAI poultry outbreaks in 2004 alone, and 1 confirmed and 2 likely other human HPAI infections between 2004 and 2006. METHODS: In 2008, we enrolled a cohort of 800 rural Thai adults living in 8 sites within Kamphaeng Phet Province in a prospective study of zoonotic influenza transmission. We studied participants' sera with serologic assays against 16 avian, 2 swine, and 8 human influenza viruses. RESULTS: Among participants (mean age 49.6 years and 58% female) 65% reported lifetime poultry exposure of at least 30 consecutive minutes. Enrollees had elevated antibodies by microneutralization assay against 3 avian viruses: A/Hong Kong/1073/1999(H9N2), A/Thailand/676/2005(H5N1), and A/Thailand/384/2006(H5N1). Bivariate risk factor modeling demonstrated that male gender, lack of an indoor water source, and tobacco use were associated with elevated titers against avian H9N2 virus. Multivariate modeling suggested that increasing age, lack of an indoor water source, and chronic breathing problems were associated with infection with 1 or both HPAI H5N1 strains. Poultry exposure was not associated with positive serologic findings. CONCLUSIONS: These data suggest that people in rural central Thailand may have experienced subclinical avian influenza infections as a result of yet unidentified environmental exposures. Lack of an indoor water source may play a role in transmission.

Response of feral cats to vaccination at the time of neutering.

OBJECTIVE: To determine whether administration of inactivated virus or modified-live virus (MLV) vaccines to feral cats at the time of neutering induces protective serum antiviral antibody titers. DESIGN: Prospective study. ANIMALS: 61 feral cats included in a trap-neuter-return program in Florida. PROCEDURES: Each cat received vaccines against feline panleukopenia virus (FPV), feline herpes virus (FHV), feline calicivirus (FCV), FeLV, and rabies virus (RV). Immediately on completion of surgery, vaccines that contained inactivated RV and FeLV antigens and either MLV or inactivated FPV, FHV, and FCV antigens were administered. Titers of antiviral antibodies (except those against FeLV) were assessed in serum samples obtained immediately prior to surgery and approximately 10 weeks later. RESULTS: Prior to vaccination, some of the cats had protective serum antibody titers against FPV (33%), FHV (21%), FCV (64%), and RV (3%). Following vaccination, the overall proportion of cats with protective serum antiviral antibody titers increased (FPV [90%], FHV [56%], FCV [93%], and RV [98%]). With the exception of the FHV vaccine, there were no differences in the proportions of cats protected with inactivated virus versus MLV vaccines. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that exposure to FPV, FHV, and FCV is common among feral cats and that a high proportion of cats are susceptible to RV infection. Feral cats appeared to have an excellent immune response following vaccination at the time of neutering. Incorporation of vaccination into trap-neuter-return programs is likely to protect the health of individual cats and possibly reduce the disease burden in the community.

High rate of A(H1N1)pdm09 infections among rural Thai villagers, 2009-2010.

BACKGROUND: Pandemic influenza A(H1N1)pdm09 emerged in Thailand in 2009. A prospective longitudinal adult cohort and household transmission study of influenza-like illness (ILI) was ongoing in rural Thailand at the time of emergence. Symptomatic and subclinical A(H1N1)pdm09 infection rates in the cohort and among household members were evaluated. METHODS: A cohort of 800 Thai adults underwent active community-based surveillance for ILI from 2008-2010. Acute respiratory samples from ILI episodes were tested for A(H1N1)pdm09 by qRT-PCR; acute and 60-day convalescent blood samples were tested by A(H1N1)pdm09 hemagglutination inhibition assay (HI). Enrollment, 12-month and 24-month follow-up blood samples were tested for A(H1N1)pdm09 seroconversion by HI. Household members of influenza A-infected cohort subjects with ILI were enrolled in household transmission investigations in which day 0 and 60 blood samples and acute respiratory samples were tested by either qRT-PCR or HI for A(H1N1)pdm09. Seroconversion between annual blood samples without A(H1N1)pdm09-positive ILI was considered as subclinical infection. RESULTS: The 2-yr cumulative incidence of A(H1N1)pdm09 infection in the cohort in 2009/2010 was 10.8% (84/781) with an annual incidence of 1.2% in 2009 and 9.7% in 2010; 83.3% of infections were subclinical (50% in 2009 and 85.9% in 2010). The 2-yr cumulative incidence was lowest (5%) in adults born ≤ 1957. The A(H1N1)pdm09 secondary attack rate among household contacts was 47.2% (17/36); 47.1% of these infections were subclinical. The highest A(H1N1)pdm09 secondary attack rate among household contacts (70.6%, 12/17) occurred among children born between 1990 and 2003. CONCLUSION: Subclinical A(H1N1)pdm09 infections in Thai adults occurred frequently and accounted for a greater proportion of all A(H1N1)pdm09 infections than previously estimated. The role of subclinical infections in A(H1N1)pdm09 transmission has important implications in formulating strategies to predict and prevent the spread of A(H1N1)pdm09 and other influenza virus strains.

Performance of the 10-item Center for Epidemiologic Studies Depression scale for caregiving research.

OBJECTIVES: The Center for Epidemiologic Studies Depression (CESD) scale has been useful in a broad spectrum of health research on patient and population outcomes. A brief version is used when depressive symptoms are not the primary focus. Rasch (item response) analysis previously demonstrated potential problems with positively worded items. We tested the 10-item CESD (CESD-10) scale and considered an 8-item version with both psychometric and Rasch analyses. METHODS: This was a special sample of 2067 caregivers from three existing US databases. We describe item response patterns and internal constancy in addition to Rasch scale results. RESULTS: There were few problems with missing data, and internal consistency was high (alpha = 0.86-0.88) for both CESD versions. Rasch analysis indicated that one of the positive items ("hopeful about future") could be dropped. CONCLUSIONS: We partly confirmed prior work that suggested dropping positive items for the CESD-10. Among caregivers, item-level problems and scaling problems seem minimal. At present, there is not a strong rationale for dropping the CESD-10 positive items: the one poorly performing positive item might be explained by the special caregiver sample.

Recovery of live virus after storage at ambient temperature using ViveST™.

BACKGROUND: A major impediment to performing virological field studies in developing nations is the lack of ultra-low freezers as well as the expense and difficulty of shipping frozen samples. A commercially available product, ViveST™, was developed to preserve nucleic acids at ambient temperature for use in specimen storage and transportation. However, its applications as a viral storage, transport and recovery device have not been evaluated. OBJECTIVE: To examine the ability of ViveST to preserve live virus following storage at ambient temperature. STUDY DESIGN: A panel of six viruses was stored at ambient temperature (~22°C) in ViveST with fetal bovine serum (FBS), or ViveST with minimal essential media (MEM) and compared with virus stored in universal transport media (M4RT), MEM, and FBS alone. Stored viruses included: human adenovirus (14p), dengue virus 2 (16608), echovirus 3 (Morrisey), human rhinovirus 15 (1734), Coxsackie virus B5 (Faulkner), and herpes simplex virus 1 (HF). After 7 days storage at ambient temperature, virus recovery was measured via titration using viral plaque assays or focus-forming unit assays. RESULTS: Viral titer studies indicate that ViveST with either FBS or M4RT preserved/recovered 5 different viruses for 1 week at ambient temperature. MEM preserved 4 viruses while FBS and ViveST with MEM preserved 3 viruses each. Statistical analyses indicate that M4RT and ViveST with FBS preserved significantly more virus than the other treatments. CONCLUSIONS: These data suggest that ViveST with either FBS or M4RT may be useful in field specimen collection scenarios where ultra-cold storage is not available.

Isolation and characterization of H3N8 equine influenza A virus associated with the 2011 epizootic in Mongolia.

BACKGROUND: Equine influenza virus (EIV) epizootics affect 2.1 million Mongolian horses approximately every 10 years and critically impact economy and nomadic livelihood of Mongolia. OBJECTIVES: An active surveillance program was established in 2011 to monitor influenza viruses circulating among Mongolian horses. METHODS: Nasal swabs were collected from horses in free-ranging horse herds in Töv, Khentii, and Dundgovi aimags (provinces) from January to September 2011. Real-time reversetranscriptase-polymerase chain reaction (rRT-PCR) was used to determine the presence of influenza A virus. Influenza A-positive specimens were cultured to amplify virus; viral RNA was extracted, and gene segments were amplified and sequenced by Sanger sequencing. RESULTS: A total of 745 horses were swabbed; most horses were without clinical signs of illness. In July 2011, reports of influenza-like illnesses emerged among horses in Mongolia's capital, and subsequently, surveillance efforts were adjusted to swab horses associated with the epizootic. Thirty-four specimens of rRT-PCR influenza-positive virus were collected in May, June, August, and September. Three specimens yielded detectable virus. Gene sequence studies suggested that all three isolates were identical H3N8 viruses. Phylogenetic analyses indicated the strain was very similar to other H3N8 EIVs circulating in central Asia between 2007 and 2008. CONCLUSIONS: As large Mongolian equine herds often seem to suffer from EIV epizootics, it seems prudent to continue such routine equine influenza surveillance. Doing so will provide an early warning system, should novel viruses emerge, help in assessing if EIV is crossing over to infect humans and provide data to assess the likely effectiveness of current EIV vaccines.

Serological evidence for avian H9N2 influenza virus infections among Romanian agriculture workers.

In recent years, wild birds have introduced multiple highly pathogenic avian influenza (HPAI) H5N1 virus infections in Romanian poultry. In 2005 HPAI infections were widespread among domestic poultry and anecdotal reports suggested domestic pigs may also have been exposed. We sought to examine evidence for zoonotic influenza infections among Romanian agriculture workers. Between 2009 and 2010, 363 adult participants were enrolled in a cross-sectional, seroepidemiological study. Confined animal feeding operation (CAFO) swine workers in Tulcea and small, traditional backyard farmers in Cluj-Napoca were enrolled, as well as a non-animal exposed control group from Cluj-Napoca. Enrollment sera were examined for serological evidence of previous infection with 9 avian and 3 human influenza virus strains. Serologic assays showed no evidence of previous infection with 7 low pathogenic avian influenza viruses or with HPAI H5N1. However, 33 participants (9.1%) had elevated microneutralization antibody titers against avian-like A/Hong Kong/1073/1999(H9N2), 5 with titers ≥ 1:80 whom all reported exposure to poultry. Moderate poultry exposure was significantly associated with elevated titers after controlling for the subjects' age (adjusted OR = 3.6; 95% CI, 1.1-12.1). There was no evidence that previous infection with human H3N2 or H2N2 viruses were confounding the H9N2 seroreactivity. These data suggest that H9N2 virus may have circulated in Romanian poultry and occasionally infected man.

Evidence for avian H9N2 influenza virus infections among rural villagers in Cambodia.

BACKGROUND: Southeast Asia remains a critical region for the emergence of novel and/or zoonotic influenza, underscoring the importance of extensive sampling in rural areas where early transmission is most likely to occur. METHODS: In 2008, 800 adult participants from eight sites were enrolled in a prospective population-based study of avian influenza (AI) virus transmission where highly pathogenic avian influenza (HPAI) H5N1 virus had been reported in humans and poultry from 2006 to 2008. From their enrollment sera and questionnaires, we report risk factor findings for serologic evidence of previous infection with 18 AI virus strains. RESULTS: Serologic assays revealed no evidence of previous infection with 13 different low-pathogenic AI viruses or with HPAI avian-like A/Cambodia/R0404050/2007(H5N1). However, 21 participants had elevated antibodies against avian-like A/Hong Kong/1073/1999(H9N2), validated with a monoclonal antibody blocking ELISA assay specific for avian H9. CONCLUSIONS: Although cross-reaction from antibodies against human influenza viruses cannot be completely excluded, the study data suggest that a number of participants were previously infected with the avian-like A/Hong Kong/1073/1999(H9N2) virus, likely due to as yet unidentified environmental exposures. Prospective data from this cohort will help us better understand the serology of zoonotic influenza infection in a rural cohort in SE Asia.

A comparison of viral fitness and virulence between emergent adenovirus 14p1 and prototype adenovirus 14p strains.

BACKGROUND: Epidemiological studies from the last decade have suggested that the morbidity and mortality associated with a newly emergent strain of human adenovirus (HAdV-14p1) is greater than other, more prevalent, adenovirus strains. Recent molecular analysis identified very minor genetic differences in HAdV-14p1 compared to prototype HAdV-14p. No studies have evaluated how these differences may affect virulence. OBJECTIVE: To compare HAdV-14p1 and HAdV-14p strains for competitive fitness and virulence. STUDY DESIGN: We performed in vitro and molecular assays to evaluate growth kinetics, cellular infectivity, cytotoxicity, and plaque morphology of the two strains. RESULTS: Growth kinetic data showed no viral replication at 30°C and minimal differences at 37°C for both strains. Cellular infectivity data showed propagation capabilities for both strains in a diverse array of cell lines, with human lung and kidney cells having the highest propagation potential. Cytotoxicity data indicated cellular distress differences induced by both strains of virus in the first 12h, but similar distress levels between 12 and 48 h. Plaque morphology assays showed some differences in average plaque diameter. CONCLUSIONS: These data suggest that the increase in morbidity and mortality observed in recent HAdV-14p1 infections is not due to viral growth or cellular infectivity differences from the prototypic HAdV-14 strain. While there were some statistically important differences detected between strains in cytotoxicity and plaque morphology assays, it seems more likely that other factors, such as environmental stressors, co-infections, or individual host response are likely contributing to the increase in morbidity.

Long-term fertility control in female cats with GonaCon™, a GnRH immunocontraceptive.

The uncontrolled reproduction of free-roaming feral cats contributes to overpopulation and associated concerns regarding their welfare and impact on public health and the environment. Nonsurgical fertility control that could be administered to feral cats in the field would be a powerful tool for cat population control. The objective was to test the efficacy and duration of activity of a single-dose GnRH immunocontraceptive vaccine (GonaCon™) on the fertility of adult female laboratory cats. Vaccinated cats (n = 15) received a single injection of vaccine containing a GnRH-KLH conjugate (200 µg) emulsified in a mycobacterial and oil adjuvant on study Day 0. Sham-treated cats (n = 5) received a single injection containing all vaccine components except the GnRH-KLH conjugate. A breeding trial started on study Day 120. Vaccinated cats had a longer time to conception (median 39.7 mo) compared to sham-treated cats (4.4 mo; P < 0.001). A total of 93% of vaccinated cats remained infertile for the first year following vaccination, whereas 73, 53, and 40% were infertile for 2, 3, and 4 y, respectively. At study termination (5 y after a single GnRH vaccine was administered), four cats (27%) remained infertile. The GnRH antibody titers declined more rapidly in short-term responding cats with < 2 y of infertility (n = 4), compared to long-term responding cats that experienced fertility control for >2 y (n = 11) (P < 0.05). Non-painful but persistent late-onset granulomatous injection site masses appeared 2 y after initial vaccination in five cats. We concluded that GnRH immunocontraception is an ideal candidate for further development for feral cat control.

A national study of US bird banders for evidence of avian influenza virus infections.

BACKGROUND: Previously we have found that Midwestern US wildlife biologists, poultry farmers, veterinarians, and duck hunters have had evidence of avian influenza virus infections (AIVs). OBJECTIVES: We sought to evaluate a national sample of US bird banders for previous evidence of AIV infection. STUDY DESIGN: Controlled, cross-sectional serological survey. RESULTS: In 2009 and 2010 we enrolled 157 registered bird banders from 40 US states and compared their enrollment data and serological results with 78 adult age-group matched controls from Iowa. On average, the bird banders had 15 years of wild bird exposure, banded 20 days per year, worked chiefly in 1 of the 4 North American flyways, and banded 300 individual birds of 5 different species per season. While handling birds, only 15% of banders reported wearing gloves. Three bird banders and 1 control had evidence of previous infection (1 AIV each) with A/BWTE/Ohio/07/495762-6(H7N3), A/Ty/MN/38391-6/95(H9N2) or A/CK/NJ/7290-2/95(H11N3) by microneutralization assay. There was no evidence of previous infection with a representative sample of H4, H5, H6, H8, or H10 AIVs. Participants were followed for influenza-like-illness for a median of 7 months and 4 (3 bird banders) submitted self-collected eye, nasal, and throat influenza-like-illness swab specimens, 1 of which collected in November of 2009, yielded a pandemic H1N1 influenza A virus. CONCLUSION: Despite reports of conjunctivitis and upper respiratory symptoms while bird banding, we found sparse evidence that US bird banders had infections with AIVs.