#Respiratory disease due to mixed viral #infections in #poultry flocks in #Egypt between 2017 and 2018: #Upsurge of highly pathogenic #avian #influenza virus subtype #H5N8 since 2018 (Transbound Emerg Dis., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Transbound Emerg Dis. 2019 Jul 11. doi: 10.1111/tbed.13281. [Epub ahead of print]

Respiratory disease due to mixed viral infections in poultry flocks in Egypt between 2017 and 2018: Upsurge of highly pathogenic avian influenza virus subtype H5N8 since 2018.

Hassan KE1,2, El-Kady MF2, El-Sawah AAA2, Luttermann C3, Parvin R1,4, Shany S2, Beer M1, Harder T1.

Author information: 1 Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, Greifswald-Riems, Germany. 2 Department of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt. 3 Institute of Immunology Virology, Friedrich-Loeffler-Institute, Greifswald-Riems, Germany. 4 Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh.

 

Abstract

For several years, poultry production in Egypt has been suffering from co-circulation of multiple respiratory viruses including highly pathogenic avian influenza virus (HPAIV) H5N1 (clade 2.2.1.2) and low pathogenic H9N2 (clade G1-B). Incursion of HPAIV H5N8 (clade 2.3.4.4b) to Egypt in November 2016 via wild birds followed by spread into commercial poultry flocks further complicated the situation. Current analyses focussed on 39 poultry farms suffering from respiratory manifestation and high mortality in six Egyptian governorates during 2017-2018. Real-time RT-PCR (RT-qPCR) substantiated the co-presence of at least two respiratory virus species in more than 80% of the investigated flocks. The percentage of HPAIV H5N1-positive holdings was fairly stable in 2017 (12.8%) and 2018 (10.2%), while the percentage of HPAIV H5N8-positive holdings increased from 23% in 2017 to 66.6% during 2018. The proportion of H9N2-positive samples was constantly high (2017:100% and 2018:63%), and H9N2 co-circulated with HPAIV H5N8 in 22 out of 39 (56.8%) flocks. Analyses of 26 H5, 18 H9 and 4 N2 new sequences confirmed continuous genetic diversification. In silico analysis revealed numerous amino acid substitutions in the HA and NA proteins suggestive of increased adaptation to mammalian hosts and putative antigenic variation. For sensitive detection of H9N2 viruses by RT-qPCR, an update of primers and probe sequences was crucial. Reasons for the relative increase of HPAIV H5N8 infections versus H5N1 remained unclear, but lack of suitable vaccines against clade 2.3.4.4b cannot be excluded. A reconsideration of surveillance and control measures should include updating of diagnostic tools and vaccination strategies.

© 2019 Blackwell Verlag GmbH.

KEYWORDS: Egypt; Highly pathogenic avian influenza; co-infection; control; diagnostic tools; reassortant viruses

PMID: 31297991 DOI: 10.1111/tbed.13281

Keywords: Avian Influenza; H5N1; H5N8; H9N2; Poultry; Egypt.

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Serial Section Array Scanning Electron Microscopy Analysis of Cells from #Lung #Autopsy #Specimens Following #Fatal A/ #H1N1pdm09 #Influenza Virus #Infection (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Serial Section Array Scanning Electron Microscopy Analysis of Cells from Lung Autopsy Specimens Following Fatal A/H1N1 2009 Pandemic Influenza Virus Infection

Michiyo Kataoka, Kinji Ishida, Katsutoshi Ogasawara, Takayuki Nozaki, Yoh-ichi Satoh, Tetsutaro Sata, Yuko Sato, Hideki Hasegawa, Noriko Nakajima

DOI: 10.1128/JVI.00644-19

 

ABSTRACT

A/H1N1 2009 pandemic influenza virus (A/H1N1/pdm09) was first identified as a novel pandemic influenza A virus (IAV) in 2009. Previously, we reported that many viral antigens were detected in type II alveolar epithelial cells (AEC-IIs) within autopsied lung tissue from a patient with A/H1N1/pdm09 pneumonia. It is important to identify the association between the virus and host cells to elucidate the pathogenesis of IAV pneumonia. To investigate the distribution of virus particles and morphological changes in host cells, the autopsied lung specimens from this patient were examined using transmission electron microscopy (TEM) and a novel scanning electron microscopy (SEM) method. We focused on AEC-IIs as viral antigen-positive cells, and on monocytes/macrophages (Ms/MΦs) and neutrophils (Neus) as innate immune cells. We identified virus particles and intranuclear dense tubules, which are associated with matrix 1 (M1) proteins from IAV. Large-scale two-dimensional observation was enabled by digitally ‘stitching’ together contiguous SEM images. A single whole cell analysis using a serial section array (SSA)-SEM identified virus particles in vesicles within the cytoplasm and/or around the cell surface of AEC-IIs, Ms/MΦs, and Neus; however, intranuclear dense tubules were found only in AEC-IIs. Computer-assisted processing of SSA-SEM images from each cell type enabled 3D modeling of the distribution of virus particles within an ACE-II, a M/MΦ, and a Neu.

 

IMPORTANCE

Generally, it is difficult to observe IAV particles in post-mortem samples from patients with seasonal influenza. In fact, only a few viral antigens are detected in bronchial epithelial cells from autopsied lung sections. Previously, we detected many viral antigens in AEC-IIs from the lung. This was because the majority of A/H1N1/pdm09 in the lung tissue harbored an aspartic acid to glycine substitution at position 222 (D222G) of the hemagglutinin protein. A/H1N1/pdm09 harboring the D222G substitution has a receptor-binding preference for α-2,3-linked sialic acids expressed on human AECs and infects them in the same way as H5N1 and H7N9 avian IAVs. Here, we report the first successful observation of virus particles not only in AEC-IIs, but also in Ms/MΦs and Neus, using electron microscopy. The finding of a M/MΦ harboring numerous virus particles within vesicles and at the cell surface suggests that Ms/MΦs are involved in the pathogenesis of IAV primary pneumonia.

Copyright © 2019 Kataoka et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

Keywords: Influenza A; Seasonal Influenza; Pandemic Influenza; Avian Influenza; H1N1pdm09; H5N1; H7N9; Viral pathogenesis.

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A novel I117T #substitution in #neuraminidase of highly pathogenic #avian #influenza #H5N1 virus conferring reduced susceptibility to #oseltamivir and #zanamivir (Vet Microbiol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Vet Microbiol. 2019 Jun 5;235:21-24. doi: 10.1016/j.vetmic.2019.06.005. [Epub ahead of print]

A novel I117T substitution in neuraminidase of highly pathogenic avian influenza H5N1 virus conferring reduced susceptibility to oseltamivir and zanamivir.

Kode SS1, Pawar SD2, Tare DS1, Keng SS1, Hurt AC3, Mullick J1.

Author information: 1 Avian Influenza Group, Microbial Containment Complex, ICMR-National Institute of Virology, 130/1, Sus Road, Pashan, Pune 411021, India. 2 Avian Influenza Group, Microbial Containment Complex, ICMR-National Institute of Virology, 130/1, Sus Road, Pashan, Pune 411021, India. Electronic address: shaileshpawarniv@gmail.com. 3 WHO Collaborating Centre for Reference and Research on Influenza (VIDRL), Peter Doherty Institute for Infection and Immunity, 792 Elizabeth Street, Melbourne, VIC 3000, Australia.

 

Abstract

Occurrence of avian influenza (AI) with Neuraminidase (NA) mutations which confer reduced neuraminidase inhibitor (NAI) susceptibility has remained a cause of concern. The susceptibility to NAIs of 67 highly pathogenic avian influenza H5N1 viruses isolated during 2006-2012 in India was tested in phenotypic fluorescence-based NA inhibition assay, sequence analysis and in ovo. One isolate showed a novel NA I117T amino acid substitution (N2 numbering) and eight isolates showed previously known NAI-resistance marker mutations (I117V, E119D, N294S, total 9/67). The overall incidence of resistant variants was 13.4%. The novel I117T substitution reduced oseltamivir susceptibility by 18.6-fold and zanamivir susceptibility by 11.8-fold, compared to the wild type AI H5N1virus, thus showed cross-resistance to both oseltamivir and zanamivir in NA inhibition assays. However, the other two isolates with I117V substitution were sensitive to both the NAIs. In addition, the comparison of growth of the I117T and I117V variants in presence of NAI’s in the in ovo assays exhibited difference in growth levels. The present study reports the natural occurrence of a novel I117T mutation in AI H5N1 virus conferring cross-resistance to oseltamivir and zanamivir highlighting the urgent need of antiviral surveillance of AI viruses.

Copyright © 2019 Elsevier B.V. All rights reserved.

KEYWORDS: Antiviral resistance; H5N1; I117T substitution; In ovo assay; Oseltamivir; Zanamivir

PMID: 31282375 DOI: 10.1016/j.vetmic.2019.06.005

Keywords: Avian Influenza; H5N1; Antivirals; Drugs Resistance; Oseltamivir; Zanamivir; India.

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Evidence of #exposure and #human #seroconversion during an #outbreak of #avian #influenza A(#H5N1) among #poultry in #Cameroon (Emerg Microbes Infect., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Emerg Microbes Infect. 2019;8(1):186-196. doi: 10.1080/22221751.2018.1564631.

Evidence of exposure and human seroconversion during an outbreak of avian influenza A(H5N1) among poultry in Cameroon.

Monamele CG1, Y P2, Karlsson EA2, Vernet MA1, Wade A3, Okomo MA4, Abah ASA5, Yann S2, Etoundi GAM5, Mohamadou NR1, Feussom JM3, Horm S2, Horwood PF2,6, Ly S7, Njouom R1, Dussart P2.

Author information: 1a Virology Department, Centre Pasteur du Cameroun , Institut Pasteur International Network , Yaoundé , Cameroon. 2b Virology Unit , Institut Pasteur du Cambodge, Institut Pasteur International Network , Phnom Penh , Cambodia. 3c Ministry of Livestock, Fisheries and Animal Industries , Yaoundé , Cameroon. 4d National Laboratory of Public Health , Yaoundé , Cameroon. 5e Ministry of Public Health , Yaoundé , Cameroon. 6f Australian Institute of Tropical Health and Medicine , James Cook University , Cairns , Australia. 7g Epidemiology and Public Health Unit , Institut Pasteur du Cambodge, Institut Pasteur International Network , Phnom Penh , Cambodia.

 

Abstract

From May 2016 to March 2017, 22 poultry outbreaks of avian influenza A(H5N1) were reported in Cameroon, mainly in poultry farms and live bird markets. No human cases were reported. In this study, we sought to describe the 2016 A(H5N1) outbreak strain and to investigate the risk of infection in exposed individuals. We find that highly pathogenic influenza subtype A(H5N1), clade 2.3.2.1c from Cameroon is closely related phylogenetically and antigenically to strains isolated in central and western Africa at the time. No molecular markers of increased human transmissibility were noted; however, seroconversion was detected in two poultry workers (1.5% of total screened). Therefore, the continued outbreaks of avian influenza in poultry and the risk of zoonotic human infection highlight the crucial need for continued and vigilant influenza surveillance and research in Africa, especially in areas of high poultry trade, such as Cameroon.

KEYWORDS: Africa; Avian influenza; Cameroon; H5N1; Zoonoses; outbreak; surveillance

PMID: 30866772 PMCID: PMC6455145 DOI: 10.1080/22221751.2018.1564631 [Indexed for MEDLINE] Free PMC Article

Keywords: Avian Influenza; H5N1; Seroprevalence; Human; Poultry; Cameroon.

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The #safety and #immunogenicity of a cell-derived adjuvanted #H5N1 #vaccine – A phase I randomized clinical trial (J Microbiol Immunol Infect., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

J Microbiol Immunol Infect. 2019 May 18. pii: S1684-1182(18)30176-2. doi: 10.1016/j.jmii.2019.03.009. [Epub ahead of print]

The safety and immunogenicity of a cell-derived adjuvanted H5N1 vaccine – A phase I randomized clinical trial.

Cheng A1, Hsieh SM1, Pan SC1, Li YH2, Hsieh EF2, Lee HC2, Lin TW2, Lai KL3, Chen C2, Shi-Chung Chang S3, Chang SC4.

Author information: 1 Department of Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan. 2 Medigen Vaccine Biologics Corporation, Hsinchu, Taiwan. 3 Medigen Biotechnology Corporation, Taipei, Taiwan. 4 Department of Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan. Electronic address: changsc@ntu.edu.tw.

 

Abstract

BACKGROUND:

Development of an efficacious egg-free mock-up H5N1 vaccine is key to our preparedness against pandemic avian flu.

METHODS:

This is a single-center, randomized, observer-blinded phase I clinical trial evaluating the safety and immunogenicity of an alum-adjuvanted Madin-Darby canine kidney (MDCK)-derived inactivated whole-virion H5N1 influenza vaccine in healthy adults. Hemagglutination inhibition (HAI) and neutralizing antibody titers were measured using horse and turkey red blood cells (RBCs).

RESULTS:

Thirty-six adult subjects were randomized to receive two doses of 0.5 mL of the MDCK-derived H5N1 alum-adjuvanted vaccine containing 7.5, 15, or 30 μg of hemagglutinin (HA) 21 days apart. The candidate vaccine was well tolerated and safe across the three dosing groups. The most frequent adverse event was injection site pain (46.5%). Both HAI and neutralizing antibody titers increased after each vaccination in all three dosing groups. The best HAI responses, namely a seroconversion rate of 91.7% and a geometric mean ratio of 9.51 were achieved with the HA dose of 30 μg assayed using horse RBCs at day 42. HAI titers against H5N1 avian influenza virus was significantly higher when measured using horse RBCs compared with turkey RBCs.

CONCLUSIONS:

This Phase I trial showed the MDCK-derived H5N1 candidate vaccine is safe and immunogenic. The source of RBCs has a significant impact on the measurement of HAI titers (ClinicalTrials.gov number: NCT01675284.).

Copyright © 2019. Published by Elsevier B.V.

KEYWORDS: Avian influenza; H5N1; Immune response; Influenza A; Vaccination

PMID: 31255574 DOI: 10.1016/j.jmii.2019.03.009

Keywords: Avian Influenza; H5N1; Vaccines.

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Identification and #analysis of long non-coding #RNAs in response to #H5N1 #influenza viruses in #duck (Anas platyrhynchos) (BMC Genomics, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

BMC Genomics. 2019 Jan 11;20(1):36. doi: 10.1186/s12864-018-5422-2.

Identification and analysis of long non-coding RNAs in response to H5N1 influenza viruses in duck (Anas platyrhynchos).

Lu C1, Xing Y1, Cai H1, Shi Y1, Liu J2, Huang Y3.

Author information: 1 State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, 100193, China. 2 Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, China. 3 State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, 100193, China. cauhyh@cau.edu.cn.

 

Abstract

BACKGROUND:

Long non-coding RNAs (lncRNAs) are important component of mammalian genomes, where their numbers are even larger than that of protein-coding genes. For example, human (Homo sapiens) (96,308 vs. 20,376) and mouse (Mus musculus) (87,774 vs. 22,630) have more lncRNA genes than protein-coding genes in the NONCODEv5 database. Recently, mammalian lncRNAs were reported to play critical roles in immune response to influenza A virus infections. Such observation inspired us to identify lncRNAs related to immune response to influenza A virus in duck, which is the most important natural host of influenza A viruses.

RESULTS:

We explored features of 62,447 lncRNAs from human, mouse, chicken, zebrafish and elegans, and developed a pipeline to identify lncRNAs using the identified features with transcriptomic data. We then collected 151,970 assembled transcripts from RNA-Seq data of 21 individuals from three tissues and annotated 4094 duck lncRNAs. Comparing to duck protein-coding transcripts, we found that 4094 lncRNAs had smaller number of exons (2.4 vs. 10.2) and longer length of transcripts (1903.0 bp vs. 1686.9 bp) on average. Among them, 3586 (87.6%) lncRNAs located in intergenic regions and 619 lncRNAs showed differential expression in ducks infected by H5N1 virus when compared to control individuals. 58 lncRNAs were involved into two co-expressional modules related to anti-influenza A virus immune response. Moreover, we confirmed that eight lncRNAs showed remarkably differential expression both in vivo (duck individuals) and in vitro (duck embryo fibroblast cells, DEF cells) after infected with H5N1 viruses, implying they might play important roles in response to influenza A virus infection.

CONCLUSIONS:

This study presented an example to annotate lncRNA in new species based on model species using transcriptome data. These data and analysis provide information for duck lncRNAs’ function in immune response to influenza A virus.

KEYWORDS: Avian H5N1 influenza virus; Duck; Immune response; LncRNAs

PMID: 30634898 PMCID: PMC6330444 DOI: 10.1186/s12864-018-5422-2 [Indexed for MEDLINE] Free PMC Article

Keywords: Avian Influenza; H5N1; Poultry.

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Development of #American-Lineage #Influenza #H5N2 #Reassortant #Vaccine Viruses for #Pandemic #Preparedness (Viruses, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Viruses. 2019 Jun 11;11(6). pii: E543. doi: 10.3390/v11060543.

Development of American-Lineage Influenza H5N2 Reassortant Vaccine Viruses for Pandemic Preparedness.

Chen PL1,2, Hu AY3, Lin CY4, Weng TC5, Lai CC6,7, Tseng YF8, Cheng MC9,10, Chia MY11,12, Lin WC13, Yeh CT14, Su IJ15, Lee MS16.

Author information: 1 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. letitia@nhri.org.tw. 2 Institute of Molecular and Cellular Biology, National Tsing Hua University, Hsinchu 30013, Taiwan. letitia@nhri.org.tw. 3 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. alanhu@nhri.org.tw. 4 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. grayingaries@outlook.com. 5 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. wtc@nhri.org.tw. 6 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. laicc2@nhri.org.tw. 7 College of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan. laicc2@nhri.org.tw. 8 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. yufents@gmail.com. 9 Department of Veterinary Medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan. mccheng@mail.npust.edu.tw. 10 Animal Health Research Institutes, Danshui, New Taipei City 25158, Taiwan. mccheng@mail.npust.edu.tw. 11 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. chiaminyuan@dragon.nchu.edu.tw. 12 Department of Veterinary Medicine, National Chung Hsing University, Taichung 40227, Taiwan. chiaminyuan@dragon.nchu.edu.tw. 13 Institute of Preventive Medicine, National Defence Medical Centre, Taipei 23742, Taiwan. spps057@gmail.com. 14 Institute of Preventive Medicine, National Defence Medical Centre, Taipei 23742, Taiwan. yyhome@mail.ndmctsgh.edu.tw. 15 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. suihjen0704@stust.edu.tw. 16 National Institution of Infectious Diseases and Vaccinology, National Health Research Institutes (NHRI), Zhunan, Miaoli 35053, Taiwan. minshi@nhri.org.tw.

 

Abstract

Novel low-pathogenic avian influenza (LPAI) H5N2 viruses hit poultry farms in Taiwan in 2003, and evolved into highly pathogenic avian influenza (HPAI) viruses in 2010. These viruses are reassortant viruses containing HA and NA genes from American-lineage H5N2 and six internal genes from local H6N1 viruses. According to a serological survey, the Taiwan H5N2 viruses can cause asymptomatic infections in poultry workers. Therefore, a development of influenza H5N2 vaccines is desirable for pandemic preparation. In this study, we employed reverse genetics to generate a vaccine virus having HA and NA genes from A/Chicken/CY/A2628/2012 (E7, LPAI) and six internal genes from a Vero cell-adapted high-growth H5N1 vaccine virus (Vero-15). The reassortant H5N2 vaccine virus, E7-V15, presented high-growth efficiency in Vero cells (512 HAU, 107.6 TCID50/mL), and passed all tests for qualification of candidate vaccine viruses. In ferret immunization, two doses of inactivated whole virus antigens (3 μg of HA protein) adjuvanted with alum could induce robust antibody response (HI titre 113.14). In conclusion, we have established reverse genetics to generate a qualified reassortant H5N2 vaccine virus for further development.

KEYWORDS: American-lineage H5N2 vaccine; American-lineage reassortant influenza viruses; Pandemic preparedness

PMID: 31212631 DOI: 10.3390/v11060543

Keywords: Avian Influenza; H5N1; H5N2; H6N1; Reassortant Strain; Vaccines.

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