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Tissue pathomorphology in myocardial and lung lesions in infected chickens, ducks and mice caused by the three H5N8 viruses (stained by H&E). (a-c) Myocardial and lung lesions from QD5, 0420 and WF1 virus-infected chickens, respectively. (d-f) Myocardial and lung lesions from QD5, 0420 and WF1 virus-infected ducks, respectively. (g-i) Myocardial and lung lesions from QD5, 0420 and WF1 virus-infected mice, respectively
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Three H5N8 avian influenza viruses isolated from domestic geese in China in 2014 were characterized phylogenetically and biologically. Phylogenetic analysis of the complete genomic sequences of the three isolates from this study and those of 61 other H5N8 viruses retrieved from the GISAID platform indicated that, chronologically and geographically,...
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Wild birds of the orders Anseriformes (mainly ducks, geese and swans) and Charadriiformes (mainly gulls, terns and waders) constitute the natural reservoir for low pathogenic avian influenza (LPAI) viruses. In Egypt, highly pathogenic avian influenza (HPAI) H5N1 and LPAI H9N2 viruses are endemic in domestic poultry, forming a threat to animal and h...
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... The selected strains' sequence analyses revealed that all H5N8 viruses were direct progeny of the K1203 (H5N8)-like viruses discovered in China in 2010 and belonged to the Asian H5N1 HA lineage of clade 2.3.4.4. The recent common clade 2.3.4.4 H5N8 reassortants, which have severely damaged the poultry sector and pose a threat to public health, were created by K1203-like viruses, according to studies (Li et al., 2014). ...
... On the basis of similarity, H5N8 viruses evolved into three groups (Li et al., 2014). Groups I and II contain the isolates belonging to clade 2.3.4.4b and the Eurasian continent, whereas group III contains isolates from the North American lineage, with apparent divergence from those in groups I and II. ...
Orthomyxoviruses are negative-sense, RNA viruses with segmented genomes that are highly unstable due to reassortment. The highly pathogenic avian influenza (HPAI) subtype H5N8 emerged in wild birds in China. Since its emergence, it has posed a significant threat to poultry and human health. Poultry meat is considered an inexpensive source of protein, but due to outbreaks of HPAI H5N8 from migratory birds in commercial flocks, the poultry meat industry has been facing severe financial crises. This review focuses on occasional epidemics that have damaged food security and poultry production across Europe, Eurasia, the Middle East, Africa, and America. HPAI H5N8 viral sequences have been retrieved from GISAID and analyzed. Virulent HPAI H5N8 belongs to clade 2.3.4.4b, Gs/GD lineage, and has been a threat to the poultry industry and the public in several countries since its first introduction. Continent-wide outbreaks have revealed that this virus is spreading globally. Thus, continuous sero- and viro-surveillance both in commercial and wild birds, and strict biosecurity reduces the risk of the HPAI virus appearing. Furthermore, homologous vaccination practices in commercial poultry need to be introduced to overcome the introduction of emergent strains. This review clearly indicates that HPAI H5N8 is a continuous threat to poultry and people and that further regional epidemiological studies are needed.
... The receptor binding characteristic of the influenza virus is that avian influenza virus preferentially binds SA α-2, 3-Gal receptor, human influenza virus preferentially binds alpha-2,6-linked sialic acid (SA α-2,6-Gal) receptor [65,66]. The ability of a virus to adapt to new hosts can be improved by the process of changing the binding properties of its receptors from preferentially binding SA α-2, 3-Gal receptor to SA α-2,6-Gal receptor. ...
In October 2021, a wild bird-origin H3N8 influenza virus-A/Chinese pond heron/Jiangxi 5-1/2021 (H3N8)-was isolated from Chinese pond heron in China. Phylogenetic and molecular analyses were performed to characterize the genetic origin of the H3N8 strain. Phylogenetic analysis revealed that eight gene segments of this avian influenza virus H3N8 belong to Eurasian lineages. HA gene clustered with avian influenza viruses is circulating in poultry in southern China. The NA gene possibly originated from wild ducks in South Korea and has the highest homology (99.3%) with A/Wild duck/South Korea/KNU2020-104/2020 (H3N8), while other internal genes have a complex and wide range of origins. The HA cleavage site is PEKQTR↓GLF with one basic amino acid, Q226 and T228 at HA preferentially bind to the alpha-2,3-linked sialic acid receptor, non-deletion of the stalk region in the NA gene and no mutations at E627K and D701N of the PB2 protein, indicating that isolate A/Chinese pond heron/Jiangxi 5-1/2021 (H3N8) was a typical avian influenza with low pathogenicity. However, there are some mutations that may increase pathogenicity and transmission in mammals, such as N30D, T215A of M1 protein, and P42S of NS1 protein. In animal studies, A/Chinese pond heron/Jiangxi 5-1/2021 (H3N8) replicates inefficiently in the mouse lung and does not adapt well to the mammalian host. Overall, A/Chinese pond heron/Jiangxi 5-1/2021 (H3N8) is a novel wild bird-origin H3N8 influenza virus reassortant from influenza viruses of poultry and wild birds. This wild bird-origin avian influenza virus is associated with wild birds along the East Asian-Australasian flyway. Therefore, surveillance of avian influenza viruses in wild birds should be strengthened to assess their mutation and pandemic risk in advance.
... The viruses were propagated in specific pathogen-free ECEs (AviFarms Pty [Ltd], Pretoria, South Africa), and EID 50 titres were determined according to the method of Reed and Muench (1938). At the time of the experiments, the stocks had been passaged three times in ECEs and the virus concentration given at 10 6 50% egg infectious doses (EID 50 )/0.1 mL, prescribed viral dose for challenge studies in avian species (Li et al. 2016). ...
... Following the increasing reports of HPAIV, a joint team of experts from WHO, FAO, and OIE developed a new classification for these viruses based on changes in the H5 gene to examine the emerging strains. The H5 viruses were categorized into ten separate phylogenetic clades (clades 0-9) based on the HA (Hemagglutinin) gene of H5 isolates from 1996 to 2006 in China [3,4].In 2010, the HPAI A/duck/ Jiangsu/k1203/2010 H5N8 virus of the Asian H5N1 lineage belonging to the 2.3.4 clade was isolated for the first time from mallard ducks at a live poultry market in the eastern region of China [5,6]. Novel H5N8 reassortments were initially isolated at live poultry markets in eastern China circa 2013, followed by wild birds and poultry in Japan and the Edited by Zhen Fu. ...
The highly pathogenic avian influenza (HPAI) H5N1 virus has received considerable attention during the past 2 decades due to its zoonotic and mutative features. This Virus is of special importance due to to the possibility of causing infection in human populations. According to it’s geographical location, Iran hosts a large number of aquatic migratory birds every year, and since these birds can be considered as the host of the H5 HPAI, the country is significantly at risk of this virus. the In this study, the molecular characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of the H5N8 strain were identified in Malard county of Tehran province and Meighan wetland of Arak city, Markazi province were investigated. Based on the analysis of the amino acid sequence of the HA genes, the cleavage site of the gene includes the PLREKRRKR/GLF polybasic amino acid motif, which is a characteristic of highly pathogenic influenza viruses. The HA gene of two viruses had T156A, S123P, S133A mutations associated with the increased mammalian sialic acid-binding, and the NA gene of two viruses had H253Y mutations associated with the resistance to antiviral drugs. Phylogenetic analysis of the HA genes indicated the classification of these viruses in the 2.3.4.4 b subclade. Although the A/Goose/Iran/180/2016 virus was also an H5N8 2.3.4.4 b virus, its cluster was separated from the A/Chicken/Iran/162/2016 virus. This means that the entry of these viruses in to the country happened through more than one window. Furthermore, it seems that the introduction of these H5N8 HPAI strains in Iran probably occurred through the West Asia–East African flyway by wild migratory aquatic birds.
... The viruses were propagated in specific pathogen-free ECEs (AviFarms Pty [Ltd], Pretoria, South Africa), and EID 50 titres were determined according to the method of Reed and Muench (1938). At the time of the experiments, the stocks had been passaged three times in ECEs and the virus concentration given at 10 6 50% egg infectious doses (EID 50 )/0.1 mL, prescribed viral dose for challenge studies in avian species (Li et al. 2016). ...
Influenza A viruses (IAVs) are typically isolated and cultured by successive passages using 9- to 11-day-old embryonated chicken eggs (ECEs) and in 14-day old ECEs for virus mutational studies. Real-time reverse transcription-polymerase chain reaction tests (RT-PCRs) are commonly used for IAV diagnosis, but virus isolation remains invaluable in terms of its high sensitivity, providing viable isolates for further studies and the ability to distinguish between viable and nonviable virus. Efforts at isolating ostrich-origin IAVs from RT-PCR positive specimens using ECEs have often been unsuccessful, raising the possibility of a species bottleneck, whereby ostrich-adapted IAVs may not readily infect and replicate in ECEs, yet the capacity of an ostrich embryo to support the replication of influenza viruses has not been previously demonstrated. This study describes an optimised method for H5 and H7 subtype IAV isolation and propagation in 28-day old embryonated ostrich eggs (EOEs), the biological equivalent of 14-day old ECEs. The viability of EOEs transported from breeding sites could be maximised by pre-incubating the eggs for 12 to 14 days prior to long-distance transportation. This method applied to studies for ostrich-adapted virus isolation and in ovo studies will enable better understanding of the virus-host interaction in ostriches and the emergence of potentially zoonotic diseases.
... Most of the outbreaks of the H5N1 in Iran took place in backyard poultry in the Mazandaran Province [1,3,7]. The Asian lineage of HPAI H5N8 A/duck/Jiangsu/k1203/2010 virus was the first to report in eastern China in 2010, isolated from mallard ducks [3,8]. H5N8 prevalences to be reported from Europe, Africa, the Middle East, and Asia in poultry and wild birds; The countries of HPAI H5N8 subtypes outbreaks are located migratory route, so H5N8 spread along the route and contaminated migratory waterfowls [3,6]. ...
Background and Aims: Global epizootic distribution of HPAI H5N8 (Clade 2.3.4.4) in poultry and wild birds was demonstrated after 2010. HPAI virus is a major concern in the birds and poultry industry and global human health. Wild migratory birds and their link to backyard birds play a critical role in spreading HPAI and creating genetic reassortment.
Materials and Methods: In this study in 2018, HPAI H5N8 was isolated from Backyard poultry (turkey) in East Azerbaijan Province, Iran. Three reports of outbreaks were submitted. The first outbreak was in a village, Sholebaran, and the second was in a Livestock market in Bahman, and the third was in another Village, Yaghbastloo. Tracheal and pancreas tissue samples were obtained from 10 dead birds, and 300 susceptible domestic birds include in the turkey and chickens. Samples' diagnosis was based on real-time reverse transcriptase PCR (RRT-PCR) and partial HA gene sequencing. Turkey samples were positive and characterized as H5N8.
Results: Phylogenetic analysis result based on a partial HA gene revealed that the Iranian HPAI H5N8 virus in our study, belong to the subgroup clade 2.3.4.4 and cluster within group B.
Conclusion: These findings indicate that it provides new insights into the evolution and spread of H5N8 in Iran; based on these results, we have to recognize an improper monitoring protocol for reducing the reassortment of them. Therefore, we could prevent HPAI from circulating.
Keywords: Phylogenetic tree; H5N8; Avian influenza; Backyard poultry; HAPI
... Every year these viruses are causing great economic loss and affecting the trade of poultry all over the world (Okwor et al., 2011;Aslam et al., 2016;Sae Silva et al., 2016). Extensive poultry farming, dense human population, lack of education, a high risk of infection and poor biosecurity measures are common factors in spread of viral diseases in Southeast Asian, North African and Middle East countries (Premnathan et al., 1992;Giasuddin et al., 2012;Lee et al., 2012;Li et al., 2016). Although extensive vaccination campaigns were introduced to control viral outbreaks but still these diseases are uncontrolled in different parts of the world especially in Pakistan. ...
Herbal medicines are becoming more popular and acceptable day by day due to their effectiveness, limited side effects, and cost-effectiveness. Cholistani plants are reported as a rich source of antibacterial, antifungal, antiprotozoal, antioxidant, and anticancer agents. The current study has evaluated antiviral potential of selected Cholistani plants. The whole plants were collected, ground and used in extract formation with n-hexane, ethyl acetate and n-butanol. All the extracts were concentrated by using a rotary evaporator and concentrate was finally dissolved in an appropriate vol of the same solvent. All of the extracts were tested for their antiviral potential by using 9-11 days old chick embryonated eggs. Each extract was tested against the Avian Influenza virus H9N2 strain (AIV), New Castle Disease virus Lasoota strain (NDV), Infectious bronchitis virus (IBV) and an Infectious bursal disease virus (IBDV). Hemagglutination test (HA) and Indirect Hemagglutination (IHA) tests were performed for different viruses. The overall order of the antiviral potential of Cholistani plants against viruses was NDV>IBV>IBDV>AIV. In terms of antiviral activity from extracts, the order of activity was n-butanol>ethyl acetate>n-hexane. The medicinal plants Achyranthes aspera, Neuroda procumbens, Panicum antidotale, Ochthochloa compressa and Suaeda fruticose were very effective against all four poultry viruses through their extracts. The low IC 50 values of these extracts confirm the high antiviral potential against these viruses. It is worth to mention that Achyranthes aspera was found positive against IBDV through all its extracts which overcome the problem of unavailability of any known drug against IBDV. In short, the study proved that Cholistani plants are rich source of antiviral agent and their extracts can be used as good source of antiviral drugs both in crude and in purified form.
... Every year these viruses are causing great economic loss and affecting the trade of poultry all over the world (Okwor et al., 2011;Aslam et al., 2016;Sae Silva et al., 2016). Extensive poultry farming, dense human population, lack of education, a high risk of infection and poor biosecurity measures are common factors in spread of viral diseases in Southeast Asian, North African and Middle East countries (Premnathan et al., 1992;Giasuddin et al., 2012;Lee et al., 2012;Li et al., 2016). Although extensive vaccination campaigns were introduced to control viral outbreaks but still these diseases are uncontrolled in different parts of the world especially in Pakistan. ...
Herbal medicines are becoming more popular and acceptable day by day due to their effectiveness, limited side effects, and cost-effectiveness. Cholistani plants are reported as a rich source of antibacterial, antifungal, antiprotozoal, antioxidant, and anticancer agents. The current study has evaluated antiviral potential of selected Cholistani plants. The whole plants were collected, ground and used in extract formation with n-hexane, ethyl acetate and n-butanol. All the extracts were concentrated by using a rotary evaporator and concentrate was finally dissolved in an appropriate vol of the same solvent. All of the extracts were tested for their antiviral potential by using 9-11 days old chick embryonated eggs. Each extract was tested against the Avian Influenza virus H9N2 strain (AIV), New Castle Disease virus Lasoota strain (NDV), Infectious bronchitis virus (IBV) and an Infectious bursal disease virus (IBDV). Hemagglutination test (HA) and Indirect Hemagglutination (IHA) tests were performed for different viruses. The overall order of the antiviral potential of Cholistani plants against viruses was NDV>IBV>IBDV>AIV. In terms of antiviral activity from extracts, the order of activity was n-butanol>ethyl acetate>n-hexane. The medicinal plants Achyranthes aspera, Neuroda procumbens, Panicum antidotale, Ochthochloa compressa and Suaeda fruticose were very effective against all four poultry viruses through their extracts. The low IC50 values of these extracts confirm the high antiviral potential against these viruses. It is worth to mention that Achyranthes aspera was found positive against IBDV through all its extracts which overcome the problem of unavailability of any known drug against IBDV. In short, the study proved that Cholistani plants are rich source of antiviral agent and their extracts can be used as good source of antiviral drugs both in crude and in purified form.
... The wild-type (WT) A/mallard/Huadong/S/2005(H5N1) (S) and A/ chicken/Anhui/QD1/2014(H5N1) (QD1) viruses were isolated from swab samples of infected poultry. As previously described, the S virus was genetically and antigenically homologous with the Re-5 vaccine, both belonging to clade 2.3.4 (Li et al., 2016 (Li et al., 2016). All experiments involving live viruses were conducted in authorized animal biosafety level 3 (ABSL-3) facilities at Yangzhou University. ...
... The wild-type (WT) A/mallard/Huadong/S/2005(H5N1) (S) and A/ chicken/Anhui/QD1/2014(H5N1) (QD1) viruses were isolated from swab samples of infected poultry. As previously described, the S virus was genetically and antigenically homologous with the Re-5 vaccine, both belonging to clade 2.3.4 (Li et al., 2016 (Li et al., 2016). All experiments involving live viruses were conducted in authorized animal biosafety level 3 (ABSL-3) facilities at Yangzhou University. ...
... All seven H5 viruses were characterized phylogenetically as previously documented (Li et al., 2016;Zhong et al., 2014), and their HA gene sequences were used to construct the phylogenetic tree using a distance-based neighbour-joining method in MEGA5.1 (Tamura et al., 2011). In addition, multiple sequence alignment was performed using MUSCLE (Edgar, 2004), and phylogenetic trees were constructed by the maximum-likelihood method in RAxML v7.0.4 (Stamatakis, 2006), with the GTRGAMMA model and 1,000 bootstrap replicates. ...
As one of the important control strategies for highly pathogenic avian influenza (HPAI) in China, vaccination has been implemented compulsively in poultry flocks since 2004. However, the emergence and dominance of the circulating antigenic variants require the update of vaccines periodically. In order to investigate the key molecular sites responsible for the antigenic drift, a total of 13 amino acid positions divergent between clade 2.3.4 H5 viruses and their descendent subclade 2.3.4.4 variants in or around the recognized antigenic epitopes A~E were initially identified through inspecting a comprehensive HA sequence alignment of the H5 subtype HPAI viruses. Subsequently, a panel of single‐site or multi‐site HA mutants were constructed by reverse genetics with two H5N1 viruses of S (clade 2.3.4) and QD1 (subclade 2.3.4.4) as the HA backbone to study their antigenic variations, respectively. The hemagglutination‐inhibition assay revealed an evident impact of mutations at sites 88, 156, 205, 208, 239 and 289 to the HA antigenicity, and highlighted that the amino acid substitutions located in the antigenic region B, especially the combined mutations at sites 205 and 208, were the major antigenic determinant which was also consistent with results from flow cytometry and antigenic mapping. Our findings provided more insights into the molecular mechanism of antigenic drift of the H5 subtype HPAI virus, which would be helpful for the selection of vaccine candidates and accordingly for the prevention and control of this devastating viral agent. This article is protected by copyright. All rights reserved.
... H5N1 highly pathogenic avian influenza viruses (HPAIVs) of the A/Goose/Guangdong/ 1/1996 (Gs/Gd) lineage have become panzootic in domestic birds in Eurasia and Africa, since they detect for the first time) [2]. HPAI A/ duck/Jiangsu/k1203/2010 H5N8 virus of the Asian H5N1 lineage (HA gene belonging to clade 2.3.4) was initially isolated from mallard ducks at a live-bird market in eastern China in 2010 [3,4]. ...
... This finding indicates the H5N8 viruses are continuously evolving. Five potential N-glycosylation sites of the Aghakhan NA protein were the same as H5N8 isolates of 2014 China isolates [4]. Phylogenetic trees of internal genes illustrated that the PA, NS, PB1, PB2, and M of Aghakhan are genetically more closely related to Korean H5N8 isolates of 2016 and 2017. ...
During 2014–2017 Clade 2.3.4.4 H5N8 highly pathogenic avian influenza viruses (HPAIVs) have spread worldwide. In 2016, an epidemic of HPAIV H5N8 in Iran caused mass deaths among wild birds, and several commercial poultry farms and captive bird holdings were affected and continue to experience problems. Several outbreaks were reported in 2017. One of them is related to Hooded crow (Corvus cornix) in a national park in Esfahan province in 2017. Whole genome sequencing and characterization have been done on the detected H5N8 sample. Based on HA sequencing results, it belongs to 2.3.4.4 clade, and the cleavage site is (PLREKRRKR/G). Phylogenetic analysis of the HA gene showed that the Iran 2017 H5N8 virus clustered within subgroup Russia 2016 2.3.4.4 b of group B in H5 clade 2.3.4.4 HPAIV. On the other hand, the NA gene of the virus is placed in group C of Eurasian lineage. Complete genome characterization of this virus revealed probable reassortment of the virus with East-Asian low-pathogenic influenza viruses. Furthermore, the virus possessed some phenotypic markers related to the increased potential for transmission and pathogenicity to mammals at internal segments. This study is the first full genome characterization H5N8 HPAIV in Iran. The data complete the puzzle of molecular epidemiology of H5N8 HPAIV in Iran and the region. Our study provides evidence for fast and continuing reassortment of H5 clade 2.3.4.4 viruses, that might lead to changes in virus structural and functional characteristics such as the route and method of transmission of the virus and virus infective, pathogenic and zoonotic potential.
