The prevalence of canine H3N8 influenza and human H1N1 and H3N2 influenza in dogs in Ohio was estimated by conducting serologic tests on 1,082 canine serum samples. In addition, risk factors, such as health status and age were examined. The prevalences of human H1N1, H3N2, and canine H3N8 influenzas were 4.0%, 2.4%, and 2.3%, respectively. Two samples were seropositive for two subtypes (H1N1 and H3N2; H1N1 and canine influenza virus [CIV] H3N8). Compared to healthy dogs, dogs with respiratory signs were 5.795 times more likely to be seropositive against H1N1 virus (p = 0.042). The prevalence of human flu infection increased with dog age and varied by serum collection month. The commercial enzyme-linked immunosorbent assay used in this study did not detect nucleoprotein-specific antibodies from many hemagglutination inhibition positive sera, which indicates a need for the development and validation of rapid tests for influenza screening in canine populations. In summary, we observed low exposure of dogs to CIV and human influenza viruses in Ohio but identified potential risk factors for consideration in future investigations. Our findings support the need for establishment of reliable diagnostic standards for serologic detection of influenza infection in canine species.
Animals ; Antibodies ; Cross-Sectional Studies ; Dogs* ; Enzyme-Linked Immunosorbent Assay ; Hemagglutination ; Hospitals, Animal* ; Humans ; Influenza A virus* ; Influenza A Virus, H1N1 Subtype ; Influenza, Human* ; Mass Screening ; Ohio* ; Orthomyxoviridae ; Prevalence ; Risk Factors ; Seroepidemiologic Studies* ; Serologic Tests

Rapid and accurate identification of an influenza outbreak is essential for patient care and treatment. We describe a next-generation sequencing (NGS)-based, unbiased deep sequencing method in clinical specimens to investigate an influenza outbreak. Nasopharyngeal swabs from patients were collected for molecular epidemiological analysis. Total RNA was sequenced by using the NGS technology as paired-end 250 bp reads. Total of 7 to 12 million reads were obtained. After mapping to the human reference genome, we analyzed the 3-4% of reads that originated from a non-human source. A BLAST search of the contigs reconstructed de novo revealed high sequence similarity with that of the pandemic H1N1 virus. In the phylogenetic analysis, the HA gene of our samples clustered closely with that of A/Senegal/VR785/2010(H1N1), A/Wisconsin/11/2013(H1N1), and A/Korea/01/2009(H1N1), and the NA gene of our samples clustered closely with A/Wisconsin/11/2013(H1N1). This study suggests that NGS-based unbiased sequencing can be effectively applied to investigate molecular characteristics of nosocomial influenza outbreak by using clinical specimens such as nasopharyngeal swabs.
Databases, Genetic ; Genotype ; High-Throughput Nucleotide Sequencing ; Humans ; Influenza A Virus, H1N1 Subtype/classification/*genetics/isolation & purification ; Influenza, Human/diagnosis/*virology ; Nasopharynx/*virology ; Nucleic Acid Amplification Techniques ; Phylogeny ; RNA, Viral/analysis/metabolism ; Sequence Analysis, RNA ; Viral Proteins/genetics

The influenza A (H1N1) virus, also known as swine flu is a leading cause of morbidity and mortality since 2009. There is a need to explore novel anti-viral drugs for overcoming the epidemics. Traditionally, different plant extracts of garlic, ginger, kalmegh, ajwain, green tea, turmeric, menthe, tulsi, etc. have been used as hopeful source of prevention and treatment of human influenza. The H1N1 virus contains an important glycoprotein, known as neuraminidase (NA) that is mainly responsible for initiation of viral infection and is essential for the life cycle of H1N1. It is responsible for sialic acid cleavage from glycans of the infected cell. We employed amino acid sequence of H1N1 NA to predict the tertiary structure using Phyre2 server and validated using ProCheck, ProSA, ProQ, and ERRAT server. Further, the modelled structure was docked with thirteen natural compounds of plant origin using AutoDock4.2. Most of the natural compounds showed effective inhibitory activity against H1N1 NA in binding condition. This study also highlights interaction of these natural inhibitors with amino residues of NA protein. Furthermore, among 13 natural compounds, theaflavin, found in green tea, was observed to inhibit H1N1 NA proteins strongly supported by lowest docking energy. Hence, it may be of interest to consider theaflavin for further in vitro and in vivo evaluation.
Amino Acid Sequence ; Curcuma ; Garlic ; Ginger ; Glycoproteins ; Hope ; In Vitro Techniques ; Influenza A virus ; Influenza A Virus, H1N1 Subtype ; Influenza, Human* ; Life Cycle Stages ; Molecular Docking Simulation ; Mortality ; N-Acetylneuraminic Acid ; Neuraminidase* ; Phytochemicals ; Plant Extracts ; Plants ; Polysaccharides ; Swine ; Tea

INTRODUCTIONThe pandemic caused by the H1N1 influenza virus in 2009 resulted in extensive morbidity and mortality worldwide. As the virus was a novel virus, there was limited data available on the clinical effects of the virus on children in Malaysia. Herein, we describe the clinical characteristics of children hospitalised with H1N1 influenza in a tertiary care centre; we also attempted to identify the risk factors associated with disease severity.

METHODSIn this retrospective study, we compared the characteristics of the children who were admitted into the University of Malaya Medical Centre, Malaysia, for H1N1 influenza during the pandemic with those who were admitted for seasonal influenza in 2002-2007.

RESULTSAmong the 77 children (aged ≤ 12 years) admitted to the centre due to H1N1 influenza from 1 July 2009-30 June 2010, nearly 60% were aged < 6 years and 40.3% had an underlying medical condition. The top three underlying medical conditions were bronchial asthma (14.3%), cardiac disease (10.4%) and neurological disorder (11.7%). The risk factors for severe disease were age < 2 years, underlying bronchial asthma and chronic lung disease. The three patients who died had a comorbid medical condition. The underlying cause of the deaths was acute respiratory distress syndrome or shock.

CONCLUSIONThe clinical presentation of the children infected with the pandemic (H1N1) 2009 influenza virus did not differ significantly from that of children infected with seasonal influenza. However, there were more complaints of fever, cough and vomiting in the former group.

Adolescent ; Child ; Child, Hospitalized ; statistics & numerical data ; Child, Preschool ; Disease Outbreaks ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; epidemiology ; therapy ; Malaysia ; epidemiology ; Male ; Retrospective Studies ; Risk Factors ; Seasons ; Tertiary Care Centers ; statistics & numerical data

Preparation of maternal strain A/PR/8/34 HA antiserum for influenza virus classical reassortment. A/PR/8/34 virus was digested by bromelain after inactivation and purification. 5%-20% sucrose continuous density gradient centrifugation method was used to purify HA protein. SIRD method was used to select the target protein. SDS-PAGE method was used to identified HA protein. High Immunogenic A/PR/8/34 HA protein was successfully prepared and HI titer reached 10240. High purity HA antiserum was identified by SIRD method. The key reagent in the classical reassortment of influenza virus was prepared, and the complete set of technical methods were explored, which laid the foundation for the independent research and development of seasonal influenza vaccine strains of China.
Animals ; Antibodies, Viral ; immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Hemagglutination Inhibition Tests ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Rabbits ; Reassortant Viruses ; genetics ; immunology

Child, Preschool ; Hospitalization ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; diagnosis ; epidemiology ; prevention & control ; Inpatients ; Malaysia ; Risk Factors

Surface enhanced Raman spectroscopy technology (SERS), using gold nanoparticles as a base, was developed for rapid and sensitive detection of virus strains. SERS can be used as a rapid and reliable method to distinguish the titers of viral replication. In the present study, we characterized H1N1 subtypes of influenza A virus strains in different conditions of pH or temperatures, while we analyzed data from SERS technology using gold nanoparticles as a base and cell cultures were employed to further confirm the data from virus strains. Origin8.0 was used to collect Raman spectra, smooth and homogenize data, and to contrast spectra. Our results indicated that the peaks of different virus strains in optimal environmental conditions (T=37 ℃/pH=7.2) reached ≥3 000. This criterion was verified by subsequent virological method. The present data indicate that the established SERS protocol can be used as a rapid and reliable method to distinguish the replication rate of virus, which can be further used in clinical samples.
Gold ; Hydrogen-Ion Concentration ; Influenza A Virus, H1N1 Subtype ; growth & development ; Nanoparticles ; Spectrum Analysis, Raman ; Temperature ; Virus Cultivation ; methods

To study the effect of Yinghua Pinggan granule (YHPG) against influenza A/H1N1 virus in vivo and on the immunologic function of infected mice. The intranasal influenza virus infection was adopted in ICR mouse to establish the influenza virus pneumonia model. At the 3rd and 7th day after the infection, the lung index and pathologic changes in lung tissues of mice were detected. Realtime PCR and flow cytometry were employed to observe the virus load in lung tissues and the levels of CD4+, CD8+, and CD4+/CD8+ in peripheral blood. The result showed that at the 3rd and 7th day after the infection, YHPG (15, 30 g x kg(-1)) can significant decrease in the lung index and virus load in lung tissues of mice infected with influenza virus, alleviate the pathologic changes in lung tissues, significantly increase the levels of CD4+ and CD4+/CD8+ ratio and reduce the levels of CD8+ in whole blood. This indicated that YHPG can inhibit the influenza virus replication, alleviate pulmonary damage and adjust the weak immunologic function of infected mice, with a certain therapeutic effect on mice infected by H1N1 virus in vivo.
Animals ; Antiviral Agents ; administration & dosage ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; physiology ; Influenza, Human ; drug therapy ; pathology ; virology ; Lung ; pathology ; virology ; Male ; Mice ; Mice, Inbred ICR ; Virus Replication ; drug effects

OBJECTIVETo understand the epidemiological characteristics of influenza outbreaks in China from 2005 to 2013.

METHODSThe data of influenza-like illness outbreaks involving 10 or more cases were collected through Public Health Emergency Management Information System and National Influenza Surveillance Information System in China, and the influenza outbreaks were identified according to the laboratory detection results. Descriptive epidemiological analysis was conducted to understand the type/subtype of influenza virus and outbreak time, area, place and extent.

RESULTSFrom 2005 to 2013, a total of 3 252 influenza-like illness outbreaks were reported in the mainland of China, in which 2 915 influenza outbreaks were laboratory confirmed, and influenza A (H1N1) pdm09 virus and influenza B virus were predominant. More influenza outbreaks were reported in the influenza A (H1N1) pandemic during 2009-2010. Influenza outbreaks mainly occurred during winter-spring, and less influenza outbreaks occurred in winter and summer vacations of schools. More influenza outbreaks were reported in southern provinces, accounting for 79% of the total. Influenza outbreaks mainly occurred in primary and middle schools, where 2 763 outbreaks were reported, accounting for 85% of the total. Average 30-99 people were involved in an outbreak.

CONCLUSIONA large number of influenza outbreaks occur during influenza season every year in China, the predominant virus type or subtype varies with season. Primary and middle schools are mainly affected by influenza outbreaks.

China ; epidemiology ; Disease Outbreaks ; Humans ; Influenza A Virus, H1N1 Subtype ; isolation & purification ; Influenza B virus ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Population Surveillance ; Schools ; Seasons

Google Flu Trends (GFT) was the first application of big data in the public health field. GFT was open online in 2009 and attracted worldwide attention immediately. However, GFT failed catching the 2009 pandemic H1N1 and kept overestimating the intensity of influenza-like illness in the 2012-2014 season in the United States. GFT model has been updated for three times since 2009, making its prediction bias controlled. Here, we summarized the mechanism GFT worked, the strategy GFT used to update, and its influence on public health.
Disease Outbreaks ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; Internet ; Population Surveillance ; Public Health ; Statistics as Topic ; United States