This paper reviews important aspects of infection of pigs with avian influenza viruses. Wild waterfowl are the main reservoir for influenza A viruses; other species, such as pigs, can be infected, but most avian strains are imperfectly adapted to replication and transmission in such new hosts. However, some avian-to-porcine host jumps have resulted in the emergence of stable swine influenza virus lineages, with major consequences for both animal and human health. Different categories of factors are involved in these cross-species adaptations, both epidemiological (relating to host-host interactions) and virological (relating to host-virus interactions). An understanding of the adaptation of avian influenza viruses to pigs has benefited from a number of recent studies, but more research is warranted to fully appreciate the key molecular and epidemiological factors involved in this intriguing viral host jump.

Influenza A virus in swine (IAV-S) circulating in the United States of America are phylogenetically and antigenically distinct. A human H3 hemagglutinin (HA) was introduced in the IAV-S gene pool in the late 1990s, sustained continued circulation, and evolved into five monophyletic genetic clades after 2009, H3 IVA-E. Across these phylogenetic clades, distinct antigenic clusters were identified, with three clusters (cyan, red and green) among the most frequently detected antigenic phenotypes. Although it was demonstrated that antigenic diversity of H3N2 IAV-S was associated with changes at a few amino acid positions in the head of the HA, the implications of this diversity on vaccine efficacy was not tested. Using antigenically representative H3N2 viruses, we compared whole inactivated virus (WIV) and live attenuated influenza vaccine (LAIV) vaccines for protection against challenge with antigenically distinct H3N2 viruses in pigs. WIV provided partial protection against antigenically distinct viruses, but did not prevent virus replication in the upper respiratory tract. In contrast, LAIV provided complete protection from disease and virus was not detected after challenge with antigenically distinct viruses.IMPORTANCE Due to the rapid evolution of the influenza A virus, vaccines require continuous strain updates. Additionally, the platform used to deliver the vaccine can have an impact on the breadth of protection. Currently, there are various vaccine platforms available to prevent influenza A virus infection in swine, and we experimentally tested two: adjuvanted-whole inactivated virus and live attenuated virus. When challenged with an antigenically distinct virus, adjuvanted-whole inactivated virus provided partial protection while live attenuated virus provided effective protection. Additional strategies are required to broaden the protective properties of inactivated virus vaccines given the dynamic antigenic landscape of co-circulating strains in North America, whereas live attenuated vaccines may require less frequent strain updates based on demonstrated cross-protection. Enhancing vaccine efficacy to control influenza infections in swine will help reduce the impact it has on swine production and reduce the risk of swine-to-human transmission.

Influenza A virus pandemics are rare events caused by novel viruses which have the ability to spread in susceptible human populations. With respect to H1 subtype viruses, swine H1N1 and H1N2 viruses occasionally cross the species barrier to cause human infection. Recently isolated from humans (termed variants), swine viruses were shown to display great genetic and antigenic diversity, hence posing considerable public health risk. Here, we utilized in vitro and in vivo approaches to provide characterization of H1 subtype variant viruses isolated since the 2009 pandemic and discuss the findings in context with previously studied H1 subtype human isolates. The variant viruses were well adapted to replicate in human respiratory cell line, Calu-3, and the respiratory tracts of mice and ferrets. However, with respect to HA activation pH, the variant viruses had fusion pH thresholds closer to that of most classical swine and triple reassortant H1 isolates rather than viruses that had adapted to humans. Consistent with previous observations for swine isolates, the tested variant viruses were capable of efficient transmission between co-housed ferrets but could transmit via respiratory droplets to differing degrees. Overall, this investigation demonstrates that swine H1 viruses that infected humans possess adaptations required for robust replication and, in some cases, efficient respiratory droplet transmission in a mammalian model, and therefore, need to be closely monitored for additional molecular changes that could facilitate transmission among humans. This work highlights the need for risk assessments of emerging H1 viruses as they continue to evolve and cause human infections.IMPORTANCE Influenza A virus is a continuously evolving respiratory pathogen. Endemic in swine, H1 and H3 subtype viruses sporadically cause human infections. As each zoonotic infection represents an opportunity for human adaptation, the emergence of a transmissible influenza virus to which there is little or no pre-existing immunity is an ongoing threat to public health. Recently isolated variant H1 subtype viruses were shown to display extensive genetic diversity and in many instances were antigenically distinct from seasonal vaccine strains. In this study, we provide characterization of representative H1N1v and H1N2v viruses isolated since the 2009 pandemic. Our results show that, although recent variant H1 viruses possess some adaptation markers of concern, these viruses have not fully adapted to humans and require further adaptation to present a pandemic threat. This investigation highlights the need for close monitoring of emerging variant influenza viruses for molecular changes that could facilitate efficient transmission among humans.

Reconstruction of the 1918 influenza virus has facilitated considerable advancements in our understanding of this extraordinary pandemic virus. However, the benefits of virus reconstruction are not limited to this one strain. Here, we provide an overview of laboratory studies which have evaluated the reconstructed 1918 virus, and highlight key discoveries about determinants of virulence and transmissibility associated with this virus in mammals. We further discuss recent and current pandemic threats from avian and swine reservoirs, and provide specific examples of how reconstruction of the 1918 pandemic virus has improved our ability to contextualize research employing novel and emerging strains. As influenza viruses continue to evolve and pose a threat to human health, studying past pandemic viruses is key to future preparedness efforts.

Swine influenza is a worldwide disease, which causes damage to the respiratory system of pigs. The H1N1 and H3N2 subtypes circulate mainly in the swine population of Mexico. There is evidence that new subtypes of influenza virus have evolved genetically and have been rearranged with human viruses and from other species; therefore, the aim of our study was to identify and characterize the genetic changes that have been generated in the different subtypes of the swine influenza virus in Mexican pigs. By sequencing and analyzing phylogenetically the eight segments that form the virus genome, the following subtypes were identified: H1N1, H3N2, H1N2 and H5N2; of which, a H1N1 subtype had a high genetic relationship with the human influenza virus. In addition, a H1N2 subtype related to the porcine H1N2 virus reported in the United States was identified, as well as, two other viruses of avian origin from the H5N2 subtype. Particularly for the H5N2 subtype, this is the first time that its presence has been reported in Mexican pigs. The analysis of these sequences demonstrates that in the swine population of Mexico, circulate viruses that have suffered punctual-specific mutations and rearrangements of their proteins with different subtypes, which have successfully adapted to the Mexican swine population. This article is protected by copyright. All rights reserved.

There remains today a critical need for new antiviral agents, particularly in view of the alarming increase in drug resistance and associated issues. The marine environment has been a prolific contributor towards the identification of novel therapeutic agents in the recent few decades. Added to this, glycans (or carbohydrate- or sugar-based compounds) have in very recent decades made outstanding contributions to the development of novel therapeutics. This review brings together these significant facets of modern drug discovery by presenting the reported literature on glycans derived from marine organisms that possess antiviral activity.The glycans have been grouped together based on the marine organism they were isolated from, namely, (1) bacteria, (2) chromists, (3) plants and (4) animals. For chromists, glycans are further subsectioned into Ochrophyta (brown algae), Miozoa (according to www.algaebase.org ; also called Myzozoa according to WoRMS, www.marinespecies.org) (dinoflagellates) and Bacillariophyta (diatoms). For plants, glycans are further subsectioned into Chlorophyta, Rhodophyta and Tracheophyta. Glycans isolated to date are reported as alginates, chitosan, extracellular polysaccharides, fucans (e.g. fucoidans), galactans (e.g. carrageenans), glycolipids, glycosaminoglycans, glycosides, glycosylated haemocyanin, laminarans, mannans, polysaccharides (not defined), rhamnans and xylomannans. Interestingly, many of the glycans displaying antiviral properties are sulfated.Reports indicate that marine-sourced glycans have exhibited antiviral activity against African swine fever virus, cytomegalovirus, dengue virus, Epstein-Barr virus, encephalomyocarditis virus, human immunodeficiency virus, hepatitis C virus, herpes simplex virus, human cytomegalovirus, human papilloma virus, human rhino virus, influenza virus, Japanese encephalitis virus, murine leukaemia virus, murine sarcoma virus, Newcastle disease virus, parainfluenza virus, respiratory syncytial virus, Semliki Forest virus, tobacco mosaic virus, vaccinia virus, varicella zoster virus, viral haemorrhagic septicaemia virus and vesicular stomatitis virus. Selected representative glycan structures are presented in Fig. 20.1.

Swine influenza A viruses (SIVs) causing outbreaks of acute, highly contagious respiratory disease in pigs also pose a potential threat to public health. European avian-like H1N1 (EA H1N1) SIVs are the predominant circulating viruses in pigs in China and also occasionally cause human infection. In this study, a high-growth reassortant virus (SH1/PR8), with HA and NA genes from a representative EA H1N1 isolate A/Swine/Shanghai/1/2014 (SH1) in China and six internal genes from the high-growth A/Puerto Rico/8/34 (PR8) virus, was generated by plasmid-based reverse genetics and tested as a candidate seed virus for the preparation of inactivated vaccine. The protective efficacy of inactivated SH1/PR8 was evaluated in mice and pigs challenged with wild-type SH1 virus. After primer and boost vaccination, the SH1/PR8 vaccine induced high-level hemagglutination inhibiting (HI) antibodies, IgG antibodies, and neutralization antibodies in mice and pigs. Mice and pigs in the vaccinated group showed less clinical phenomena and pathological changes than those in the unvaccinated group. In conclusion, the inactivated high-growth reassortant vaccine SH1/PR8 could induce high antibody levels and complete protection is expected against SH1 wild type SIV, and protection against heterologous EA H1N1 SIV needs further evaluation.

Primary porcine bronchial epithelial cells (PBECs) are an ideal model to study the molecular and pathogenic mechanisms of various porcine respiratory pathogens. However, the short lifespan of primary PBECs greatly limit their application. Here, we isolated and cultured primary PBECs and established immortalized PBECs by transfecting primary PBECs with the pEGFP-hTERT recombinant plasmid containing human telomerase reverse transcriptase (hTERT). Immortalized PBECs (hTERT-PBECs) retained the morphological and functional features of primary PBECs as indicated by cytokeratin 18 expression, telomerase activity assay, proliferation assays, karyotype analysis, and quantitative reverse-transcriptase polymerase chain reaction. Compared to primary PBECs, hTERT-PBECs had higher telomerase activity, extended replicative lifespan, and displayed enhanced proliferative activity. Moreover, this cell line is not transformed in vitro and does not exhibit a malignant phenotype in vivo, suggesting that it can be safely used in further studies. Besides, hTERT-PBECs were susceptible to swine influenza virus of H3N2 subtype and porcine circovirus type 2. In conclusion, the immortalized hTERT-PBECs represent a valuable in vitro model, which can be widely used in the study of porcine respiratory pathogenic infections.