Detection of avian influenza A(H7N9) virus from live poultry markets in Guangzhou, China: a surveillance report.

PURPOSE: A virologic surveillance program for A(H7N9) virus was conducted from April 15, 2013 to February 14, 2014 in Guangzhou, aiming to clarify the geographical distribution of A(H7N9) viruses among live poultry markets (LPMs) and poultry farms in Guangzhou. Virological and serological surveys of poultry workers were also conducted to evaluate the risk of poultry-to-human transmission of the A(H7N9) virus. METHODS: 36 retail LPMs, 6 wholesale LPMs and 8 poultry farms were involved in our surveillance program. About 20 live poultry and environmental samples were obtained from each surveillance site at every sampling time. Different environmental samples were collected to represent different poultry-related work activities. RT-PCR and virus culture were performed to identify the A(H7N9) virus. Hemagglutinin inhibition assay and RT-PCR were conducted to detect possible A(H7N9) infection among poultry workers. RESULTS: A total of 8900 live poultry and environmental samples were collected, of which 131(1.5%) were tested positive for A(H7N9) virus. 44.4% (16/36) of retail LPMs and 50.0% (3/6) of wholesale LPMs were confirmed to be contaminated. No positive samples was detected from poultry farms. A significant higher positive sample rate was found in environmental samples related to poultry selling (2.6%) and slaughtering (2.4%), compared to poultry holding (0.9%). Correspondingly, A(H7N9) viruses were isolated most frequently from slaughter zone. In addition, 316 poultry workers associated with the 19 contaminated-LPMs were recruited and a low seroprevalence (1.6%) of antibody against A(H7N9) virus was detected. An asymptomatic A(H7N9) infection was also identified by RT-PCR. CONCLUSIONS: Our study highlights the importance of conducting effective surveillance for A(H7N9) virus and provides evidence to support the assumption that slaughtering is the key process for the propagation of A(H7N9) virus in retail LPMs. Moreover, the ability of A(H7N9) virus to cross species barrier is proved to be still limited.

Influenza associated mortality in Southern China, 2010-2012.

OBJECTIVE: Influenza caused substantial morbidity and mortality worldwide. The mortality burden caused by influenza has been under evaluation; however, data assessing this burden have been relatively sparse in tropical or subtropical regions. We estimated influenza-associated mortality in Guangzhou, China and assessed the excess mortality due to different influenza virus subtypes. METHODS: We estimated influenza-associated excess mortality due to all-cause, pneumonia and influenza, cardiorespiratory disease and other influenza-associated diagnoses from weekly numbers of deaths and influenza surveillance data through negative binomial regression model during 2010-2012. RESULTS: Estimates derived from the model indicated that influenza resulted in 14.72 (95% confidence interval (CI), 12.12-17.31) deaths per 100,000 population per year from all-cause death among all ages group. Most deaths (84.2%) occurred among people aged ≥65 years. B virus caused 5.84 (95%CI, 4.10-7.58) deaths per 100,000 population for all-cause death, which was higher than A (H3N2) (4.89, 95%CI, 3.19-6.59) or A(H1N1)pdm09 (3.99, 95%CI, 2.32-5.66). CONCLUSIONS: Influenza is responsible for a substantial mortality especially among people aged ≥65 years and influenza B virus caused the highest influenza-associated mortality. The results highlight the need for seasonal influenza vaccination programs in subtropical areas to decrease excess mortality.

[Evaluation on the risks of H5, H7 and H9 avian influenza infections in Guangzhou: using data from the 2006-2012 avian influenza surveillance program].

OBJECTIVE: To analyze the results of avian influenza surveillance program in Guangzhou from 2006 to 2012 and to evaluate the risk of infections with H5, H7 and H9 subtypes avian influenza viruses. METHODS: Avian influenza surveillance system in Guangzhou consisted five components:serum surveillance on occupational population, environmental specimen surveillance of avian influenza virus, avian flu emergency surveillance, influenza viruses surveillance on ILI patient and surveillance on pneumonia of unknown causes. Hemagglutination inhibition test was conducted to detect the antibodies against H5, H7 and H9 while RT-PCR was used to test the nucleic acid of H5, H7 and H9 viruses. RESULTS: From 2006 to 2012, 4103 serum specimens were collected from occupational populations and the overall positive rate of H5/H7/H9 antibodies was 3.82% . The antibody positive rates for H5, H7 and H9 were 0.22% ,0.00% and 3.70% respectively. 4 serum specimens for H5 and H9 simultaneously showed antibody positive. The positive rate of H9 among occupational populations(4.21%)appeared higher than that from the control population(2.16%). 2028 specimens were collected from poultry sites and 55 samples found positive for H5 nucleic acid (positive rate:2.71%), 14 samples positive for H9 nucleic acid (positive rate:0.69%), 5 specimens, simultaneously positive for H5 and H9 nucleic acids. However, none of the samples showing H7 nucleic acid positive. From 2006 to 2012, all the tested H5/H7/H9 virus were negative from the respiratory/serum specimens among those close contacts of patients or high risk groups through the avian flu emergency surveillance program,ILI patient influenza virus surveillance programs or pneumonia of unknown causes surveillance program. CONCLUSION: Contamination of H5/H9 avian influenza virus did exist in the poultry sites in Guangzhou, especially in the wet Markets. The H5/H9 avian influenza virus caused asymptomatic infection was proved to be existed within the population exposed to the poultry, suggesting that the poultry occupational population in Guangzhou was under the risk of avian influenza virus infection.

A two-year surveillance of 2009 pandemic influenza A (H1N1) in Guangzhou, China: from pandemic to seasonal influenza?

In this two-years surveillance of 2009 pandemic influenza A (H1N1) (pH1N1) in Guangzhou, China, we reported here that the scale and duration of pH1N1 outbreaks, severe disease and fatality rates of pH1N1 patients were significantly lower or shorter in the second epidemic year (May 2010-April 2011) than those in the first epidemic year (May 2009-April 2010) (P<0.05), but similar to those of seasonal influenza (P>0.05). Similar to seasonal influenza, pre-existing chronic pulmonary diseases was a risk factor associated with fatal cases of pH1N1 influenza. Different from seasonal influenza, which occurred in spring/summer seasons annually, pH1N1 influenza mainly occurred in autumn/winter seasons in the first epidemic year, but prolonged to winter/spring season in the second epidemic year. The information suggests a tendency that the epidemics of pH1N1 influenza may probably further shift to spring/summer seasons and become a predominant subtype of seasonal influenza in coming years in Guangzhou, China.

Antibody dynamics of 2009 influenza A (H1N1) virus in infected patients and vaccinated people in China.

BACKGROUND: To evaluate the risk of the recurrence and the efficiency of the vaccination, we followed-up antibody responses in patients with the 2009 pandemic H1N1 influenza and persons who received the pandemic H1N1 vaccine in Guangzhou China. METHODS: We collected serum samples from 129 patients and 86 vaccinated persons at day 0, 15, 30, 180 after the disease onset or the vaccination, respectively. Antibody titers in these serum samples were determined by haemagglutination inhibition (HI) assay using a local isolated virus strain A/Guangdong Liwan/SWL1538/2009(H1N1). RESULTS: HI antibody positive rate of the patients increased significantly from 0% to 60% at day 15 (χ(2) = 78, P<0.001) and 100% at day 30 (χ(2) = 23, P<0.001), but decreased significantly to 52% at day 180 (χ(2) = 38, P<0.001), while that of vaccinated subjects increased from 0% to 78% at day 15 (χ(2) = 110, P<0.001) and 81% at day 30 (χ(2) = 0.32, P = 0.57), but decreased significantly to 34% at day 180 (χ(2) = 39, P<0.001). Geometric mean titers (GMT) of HI antibodies in positive samples from the patients did not change significantly between day 15 and day 30 (T = 0.92, P = 0.36), but it decreased significantly from 80 at day 30 to 52 at day 180 (T = 4.5, P<0.001). GMT of vaccinated persons increased significantly from 100 at day 15 to 193 at day 30 (T = 4.5, P<0.001), but deceased significantly to 74 at day 180 (T = 5.1, P<0.001). Compared to the patients, the vaccinated subjects showed lower seroconversion rate (χ(2) = 11, P<0.001; χ(2) = 5.9, P = 0.015), but higher GMT (T = 6.0, P<0.001; T = 3.6, P = 0.001) at day 30 and day 180, respectively. CONCLUSION: Vaccination of 2009 influenza A (H1N1) was effective. However, about half or more recovered patients and vaccinated persons might have lost sufficient immunity against the recurrence of the viral infection after half a year. Vaccination or re-vaccination may be necessary for prevention of the recurrence.

Epidemiological investigation of an outbreak of pandemic influenza A (H1N1) 2009 in a boarding school: serological analysis of 1570 cases.

BACKGROUND: A large number of 2009 pandemic influenza A (H1N1) infections were localized in school populations. OBJECTIVES: To describe the epidemiology, clinical features and risk factors associated with an outbreak that occurred at a vocational boarding school in Guangzhou, P.R. China. STUDY DESIGN: Data were collected prospectively and retrospectively through the use of on-site doctors and a post-outbreak survey and blood collection. The survey was used to confirm symptoms, and to investigate a series of flu-related factors such as dormitory conditions, health habits, vaccine history and population contact history. Blood samples were taken for serological analysis. Pandemic H1N1 infection was initially confirmed by a real-time RT-PCR assay. Following the identification of the outbreak by the Guangzhou CDC on September 4, cases were diagnosed symptomatically and retrospectively by serological analysis using the hemagglutination inhibition assay and a neutralization assay. RESULTS: The infection rate was 32% (505/1570) and the attack rate was 22.2% (349/1570). The asymptomatic infection rate was 9.9% (156/1570). Sharing a classroom (OR=2.17, 95% CI: 1.62-2.91) and dormitory space (OR=2.32, 95% CI: 1.84-2.93) was associated with higher rates of infection. Opening windows for ventilation was the only control measure that significantly protected against infection. CONCLUSION: Social isolation and quarantine should be used to prevent the spread of infection. Ventilation and a control of air flow between classrooms and dorms should be implemented as possible. School closures may be effective if implemented early.

Immunogenicity associated with the routine use of an influenza A H1N1 vaccine in health care personnel in Guangzhou, China.

We present immunogenicity data on the routine vaccination of 103 health care personnel during the 2009 H1N1 national vaccination campaign. The seroprotection rate (percentage of samples with hemagglutination inhibition titers of > or =1:40) was 83.2% at 30 days postvaccination, lower than those obtained in previously published controlled trials. Low baseline antibody levels and an increase in seroprotection in a negative-control cohort suggest that the virus remains prevalent.

[Avian influenza virus infection in people occupied in poultry fields in Guangzhou city].

OBJECTIVE: To conduct serological investigation on H5N1/H9N2/H7N7 infection among people occupied in poultry fields. METHODS: Serum samples were collected from people working in live poultry and none-poultry retailing food markets, poultry wholesaling, large-scale poultry breading factories and in small-scale farms, wide birds breeding, swine slaughtering houses and from normal population. Antibodies of H5, H9 and H7 with hemagglutination inhibition and neutralization tests were tested and analyzed. Logistic regression and chi(2) test were used. RESULTS: Among 2881 samples, 4 were positive to H5-Ab (0.14%), 146 were positive to H9-Ab (5.07%) and the prevalence of H9 among people from live poultry retailing (14.96%) was the highest. Prevalence rates of H9 were as follows: 8.90% in people working in the large-scale poultry breading factories, 6.69% in the live poultry wholesaling business, 3.75% in the wide birds breeding, 2.40% in the swine slaughtering, 2.21% in the non-poultry retailing, 1.77% in the rural poultry farmers and 2.30% in normal population. None was positive to H7-Ab among 1926 poultry workers. CONCLUSION: The H5 prevalence among people was much lower than expected, but the H9 prevalence was higher. None of the populations tested was found positive to H7-Ab. There was a higher risk of AIV infection in live poultry retailing, wholesaling and large-scale breading businesses, with the risk of live poultry retailing the highest. The longer the service length was, the higher the risk existed.

Food markets with live birds as source of avian influenza.

A patient may have been infected with highly pathogenic avian influenza virus H5N1 in Guangzhou, People's Republic of China, at a food market that had live birds. Virus genes were detected in 1 of 79 wire cages for birds at 9 markets. One of 110 persons in the poultry business at markets had neutralizing antibody against H5N1.

Monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay reveals high sensitivity of the nucleocapsid protein in acute-phase sera of severe acute respiratory syndrome patients.

Accurate and timely diagnosis of severe acute respiratory syndrome coronavirus (SARS-CoV) infection is a critical step in preventing another global outbreak. In this study, 829 serum specimens were collected from 643 patients initially reported to be infected with SARS-CoV. The sera were tested for the N protein of SARS-CoV by using an antigen capture enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies against the N protein of SARS-CoV and compared to 197 control serum samples from healthy donors and non-SARS febrile patients. The results of the N protein detection analysis were directly related to the serological analysis data. From 27 SARS patients who tested positive with the neutralization test, 100% of the 24 sera collected from 1 to 10 days after the onset of symptoms were positive for the N protein. N protein was not detected beyond day 11 in this group. The positive rates of N protein for sera collected at 1 to 5, 6 to 10, 11 to 15, and 16 to 20 days after the onset of symptoms for 414 samples from 298 serologically confirmed patients were 92.9, 69.8, 36.4, and 21.1%, respectively. For 294 sera from 248 serological test-negative patients, the rates were 25.6, 16.7, 9.3, and 0%, respectively. The N protein was not detected in 66 patients with cases of what was initially suspected to be SARS but serologically proven to be negative for SARS and in 197 serum samples from healthy donors and non-SARS febrile patients. The specificity of the assay was 100%. Furthermore, of 16 sera collected from four patients during the SARS recurrence in Guangzhou, 5 sera collected from 7 to 9 days after the onset of symptoms were positive for the N protein. N protein detection exhibited a high positive rate, 96 to 100%, between day 3 and day 5 after the onset of symptoms for 27 neutralization test-positive SARS patients and 298 serologically confirmed patients. The N protein detection rate continually decreased beginning with day 10, and N protein was not detected beyond day 19 after the onset of symptoms. In conclusion, an antigen capture ELISA reveals a high N protein detection rate in acute-phase sera of patients with SARS, which makes it useful for early diagnosis of SARS.

[Analysis on the risk factors of severe acute respiratory syndromes coronavirus infection in workers from animal markets].

OBJECTIVE: To investigate the possible risk factors of severe acute respiratory syndromes coronavirus (SARS-CoV) infection in workers from animal markets. METHODS: Self-designed questionnaires were used and serum samples were tested. Logistic regression analysis was used to analyze the data. RESULTS: Results from simple factor logistic regression analysis showed that jobs which dealing with domestic livestock, wild livestock, wild animals, aquatics were related to risk factors of SARS-CoV infection. Results from multifactor logistic regression analysis showed that jobs that dealing with wild livestock and poultry were important risk factors with OR 12.28 and 0.41. CONCLUSION: Job that dealing with palm civets was the main risk factor of SARS-CoV infection in animal market workers.

[An epidemiologic investigation on infection with severe acute respiratory syndrome coronavirus in wild animals traders in Guangzhou].

OBJECTIVE: To investigate status of infection with severe acute respiratory syndrome coronovirus (SARS-CoV) in traders of wild animals wholesale markets in Guangzhou. METHODS: Serum antibody against SARS-CoV IgG was determined cross-sectionally and symptoms of respiratory infection were investigated retrospectively for part of traders of three wholesale markets for wild animals in Guangzhou. RESULTS: Overall rate of infection with SARS-CoV in 635 traders was 16.69%, varying in three different markets. Infection rate in market A mainly engaging in wild animals ranked the highest of 25.61%, significantly higher than that in markets B and C engaging in domestic fowls and snakes. Infection rate in traders only engaging in civet cats was 58.54%, significantly higher than that in traders engaging in snakes only (9.46%). In market A, infection rate varied in different persons, 59.34%, 20.59%, 16.00%, 15.22%, 10.40% and 9.68% in traders engaging in wild animals, managers, children of the traders, traders engaging in domestic fowls, traders engaging in snakes, and traders engaging in frozen food, respectively, in a decreasing pattern as their contact opportunities. During the period of SARS epidemic, detection rate of SARS-CoV antibody in people with symptoms of acute respiratory infection was higher (30.70%) than that in those without such symptoms (20.08%). Prevalence of symptoms of acute upper respiratory infection in people with positive antibody against SARS-CoV was higher (49.28%) than that in those with negative antibody (30.35%). CONCLUSIONS: Infection with SARS-CoV in traders of animal markets possibly related to their direct exposure to wild animals, particularly to civet cats. During the period of SARS epidemic, some of the traders did infect with SARS-CoV, but they were neglected due to clinically inapparent manifestations.