Gary L. Heil

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 Systematic Review and Meta-Analysis of the Seroprevalence of Influenza A(H9N2) Infection Among Humans

INTRODUCTION Given that influenza A(H9N2) is recognized as a pandemic threat, we evaluated the overall burden of influenza A(H9N2) infections among avian-exposed human populations. METHODS We performed a systematic search of PubMed, AGRICOLA, and CAB Abstracts databases for literature published during 1997-2013. Studies reporting serological evidence of human influenza A(H9N2) infection among avian-exposed populations were included. We used a World Health Organization (WHO)-recommended case definition for serological evidence of infection based on results of hemagglutination inhibition (HI) and microneutralization (MN) assays. We calculated overall seroprevalence through a random effects meta-analysis model. RESULTS Seroprevalence data reported by the studies ranged from 1% to 43% (median, 9%) by HI, which was not significantly different from the seroprevalence estimated through the WHO-recommended case definition (median, 1.3%; range, 0.5%-42.6%). Reported seroprevalence by MN ranged from 0.6% to 9% (median, 2.7%), which was greater than the seroprevalence estimated through the WHO-recommended case definition (median, 0.3%; range, 0.1%-1.4%). CONCLUSIONS A small proportion of avian-exposed humans had evidence of influenza A(H9N2) infection. As the virus has a near global distribution in poultry, it seems likely that present surveillance efforts are missing mild or asymptomatic infections among avian-exposed persons. It seems prudent to closely monitor avian-exposed populations for influenza A(H9N2) infection to provide prepandemic warnings.

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

Equine influenza virus (EIV) epizootics affect 2·1 million Mongolian horses approximately every 10 years and critically impact economy and nomadic livelihood of Mongolia.

Influenza A(H1N1)pdm09 Virus among Healthy Show Pigs, United States

Because animals can transmit some diseases to people, it is wise to be cautious around animals that carry these diseases. But how do you know which animals are carrying disease? Sometimes they appear perfectly healthy. A study of 57 apparently healthy show pigs at a 2009 US state fair found that almost 20% were carrying influenza virus and at least 4 were carrying the 2009 pandemic virus. Of concern is the possibility that different types of influenza virus—pandemic, swine, avian—could combine in pigs and emerge as new viruses that then spread to humans. Swine workers, veterinarians, and other persons with pig contact may be at high risk for infection with pig influenza and should receive seasonal influenza vaccines, use personal protective equipment when working with healthy pigs, and limit their contact with sick pigs. Regular monitoring of influenza virus among pigs and testing of sick persons who have been exposed to pigs are needed.

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.

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. J. Med. Virol. 85:670–676, 2013.

Sparse evidence of MERS-CoV infection among animal workers living in Southern Saudi Arabia during 2012.

Middle East respiratory syndrome coronavirus (MERS‐CoV) is an emerging viral pathogen that primarily causes respiratory illness. We conducted a seroprevalence study of banked human serum samples collected in 2012 from Southern Saudi Arabia. Sera from 300 animal workers (17% with daily camel exposure) and 50 non‐animal‐exposed controls were examined for serological evidence of MERS‐CoV infection by a pseudoparticle MERS‐CoV spike protein neutralization assay. None of the sera reproducibly neutralized the MERS‐CoV‐pseudotyped lentiviral vector. These data suggest that serological evidence of zoonotic transmission of MERS‐CoV was not common among animal workers in Southern Saudi Arabia during July 2012.

Prospective Study of Avian Influenza Virus Infections among Rural Thai Villagers

Background In 2008, 800 rural Thai adults living within Kamphaeng Phet Province were enrolled in a prospective cohort study of zoonotic influenza transmission. Serological analyses of enrollment sera suggested this cohort had experienced subclinical avian influenza virus (AIV) infections with H9N2 and H5N1 viruses. Methods After enrollment, participants were contacted weekly for 24mos for acute influenza-like illnesses (ILI). Cohort members confirmed to have influenza A infections were enrolled with their household contacts in a family transmission study involving paired sera and respiratory swab collections. Cohort members also provided sera at 12 and 24 months after enrollment. Serologic and real-time RT-PCR assays were performed against avian, swine, and human influenza viruses. Results Over the 2 yrs of follow-up, 81 ILI investigations in the cohort were conducted; 31 (38%) were identified as influenza A infections by qRT-PCR. Eighty-three household contacts were enrolled; 12 (14%) reported ILIs, and 11 (92%) of those were identified as influenza infections. A number of subjects were found to have slightly elevated antibodies against avian-like A/Hong Kong/1073/1999(H9N2) virus: 21 subjects (2.7%) at 12-months and 40 subjects (5.1%) at 24-months. Among these, two largely asymptomatic acute infections with H9N2 virus were detected by >4-fold increases in annual serologic titers (final titers 1∶80). While controlling for age and influenza vaccine receipt, moderate poultry exposure was significantly associated with elevated H9N2 titers (adjusted OR = 2.3; 95% CI, 1.04–5.2) at the 24-month encounter. One subject had an elevated titer (1∶20) against H5N1 during follow-up. Conclusions From 2008–10, evidence for AIV infections was sparse among this rural population. Subclinical H9N2 AIV infections likely occurred, but serological results were confounded by antibody cross-reactions. There is a critical need for improved serological diagnostics to more accurately detect subclinical AIV infections in humans.

Sparse evidence for equine or avian influenza virus infections among Mongolian adults with animal exposures.

In recent years, Mongolia has experienced recurrent epizootics of equine influenza virus (EIV) among its 2·1 million horses and multiple incursions of highly pathogenic avian influenza (HPAI) virus via migrating birds. No human EIV or HPAI infections have been reported. In 2009, 439 adults in Mongolia were enrolled in a population‐based study of zoonotic influenza transmission. Enrollment sera were examined for serological evidence of infection with nine avian, three human, and one equine influenza virus strains. Seroreactivity was sparse among participants suggesting little human risk of zoonotic influenza infection.

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.