Swine Influenza Research Articles

Porcine Airway Organoid-Derived Well-Differentiated Epithelial Cultures as a Tool for the Characterization of Swine Influenza a Virus Strains

Swine influenza A viruses (IAVsw) are important causes of disease in pigs but also constitute a public health risk. IAVsw strains show remarkable differences in pathogenicity. We aimed to generate airway organoids from the porcine lower respiratory tract and use these to establish well-differentiated airway epithelial cell (WD-AEC) cultures grown at an air–liquid interface (ALI) for in vitro screening of IAVsw strain virulence. Epithelial cells were isolated from bronchus tissue of juvenile pigs, and airway organoids were cultured in an extracellular matrix in a culture medium containing human growth factors. Single-cell suspensions of these 3D organoids were seeded on Transwell filters and differentiated at ALI to form a pseudostratified epithelium containing ciliated cells, mucus-producing cells and tight junctions. Inoculation with a low dose of IAVsw in a low volume inoculum resulted in virus replication without requiring the addition of trypsin, and was quantified by the detection of viral genome loads in apical washes. Interestingly, inoculation of an H3N2 strain known to cause severe disease in pigs induced a greater reduction in trans-epithelial resistance and more damage to tight junctions than H1N2 or H1N1 strains associated with mild disease in pigs. We conclude that the porcine WD-AEC model is useful in assessing the virulence of IAVsw strains.

#PorkandPigs: an online media listening analysis of public perception of the U.S. swine industry

Abstract With the majority of the U.S. swine industry being generally bounded by social licensing, there is a growing need to understand social perspectives to better adapt to consumer demands. Online and social media data are rich datasets that researchers are leveraging to tackle economic and societal challenges. The information that can be gleaned from online media regarding public perceptions makes it an important tool for producers to understand driving topics of interest to the public. This study utilized a prominent online and social media listening and data analytics platform to identify and quantify online and social media mentions related to the U.S. swine industry from January 1, 2020, to December 31, 2023. A general search for online media referencing pork and pigs was designed resulting in a total of 41,093,309 mentions. The search was further queried to specific topics of food and cooking (20,580,068 mentions), price (2,866,548 mentions), welfare (1,615,208 mentions), sustainability (771,819 mentions), and top authors/organizations within the U.S. swine industry (1,430 mentions). The study spanned the geographic locations of the overall U.S. (including U.S. Minor Outlying Islands), the top 5 most populous states, and the top 5 pork producing states as of 2024. Of all mentions, X/Twitter was the largest domain for nearly all mentions related to pork and pigs. Major organizations designed to spread information between the general public and pork producers had a minor impact on the overall conversation and no online media presence in the top 5 most populous states. Net sentiment was overall positive across all four years and all geographies except for major events such as the COVID-19 pandemic, the H1N1 Swine Flu, the passing of California Proposition 12, environmental events, and inflation-associated pork prices. Overall, most media presence had positive net sentiments and with most interest surrounding ways to cook pork and how to keep bacon cheap. Understanding public perceptions of the U.S. swine industry provides the opportunity to make informed decisions on marketing strategies and production practices.

Effect of stabilizers on the detection of swine influenza A virus (swIAV) in spiked oral fluids over time.

Aggregated samples such as oral fluids (OFs) display an animal friendly and time and cost-efficient sample type for swine Influenza A virus (swIAV) monitoring. However, further molecular and biological characterization of swIAV is of particular significance. The reportedly inferior suitability of aggregated samples for subtyping of swIAV presents a major drawback compared to nasal swabs, still considered the most appropriate sample type for this purpose (Garrido-Mantilla et al. BMC Vet Res 15(1):61, 2019). In addition, the viral load in the original sample, storage conditions and characteristics of different swIAV strains might further compromise the eligibility of aggregated samples for molecular detection and subtyping. Therefore, the present study aimed to evaluate the suitability of stabilizing media to minimize the degradation of viral RNA and thus increase the detection and subtyping rate of swIAV by RT-qPCR in spiked OFs under different conditions (virus strain, storage temperature and viral load in the original sample) over a time span of 14days. The use of stabilizing media in spiked OFs resulted in a significant higher probability to detect swIAV RNA compared to OFs without stabilizers (OR = 46.1, p < 0.001). In addition,swIAV degradation over time was significantly reduced in samples suspended with stabilizer (OR = 5.80, p < 0.001), in samples stored at 4°C (OR = 2.53, p < 0.001) and in samples spiked with the avian derived H1N2 subtype (OR = 2.26, p < 0.01). No significant differences in swIAV RNA detection and degradation of swIAV RNA in spiked OFs over time were observed between the three different stabilizing media. Addition of stabilizers and storage of samples under cooled conditions significantly improved detection and subtyping of swIAV in spiked OFs.

2018-2019 human seasonal H3N2 influenza A virus spillovers into swine with demonstrated virus transmission in pigs were not sustained in the pig population.

Human seasonal H3 clade 3C3a influenza A viruses (IAV) were detected four times in U.S. pigs from commercial swine farms in Michigan, Illinois, and Virginia in 2019. To evaluate the relative risk of this spillover to the pig population, whole genome sequencing and phylogenetic characterization were conducted, and the results revealed that all eight viral gene segments were closely related to 2018-2019 H3N2 human seasonal IAV. Next, a series of in vitro viral kinetics, receptor binding, and antigenic characterization studies were performed using a representative A/swine/Virginia/A02478738/2018(H3N2) (SW/VA/19) isolate. Viral replication kinetic studies of SW/VA/19 demonstrated less efficient replication curves than all 10 swine H3N2 viruses tested but higher than three human H3N2 strains. Serial passaging experiments of SW/VA/19 in swine cells did not increase virus replication, but changes at HA amino acid positions 9 and 159 occurred. In swine transmission studies, wild-type SW/VA/19 was shed in nasal secretions and transmitted to all indirect contact pigs, whereas the human seasonal strain A/Switzerland/9715293/2013(H3N2) from the same 3C3a clade failed to transmit. SW/VA/19 induced minimal macroscopic and microscopic lung lesions. Collectively, these findings demonstrate that these human seasonal H3N2 3C3a-like viruses did not require reassortment with endemic swine IAV gene segments for virus shedding and transmission in pigs. Limited detections in the U.S. pig population in the subsequent period of time suggest a yet-unknown restriction factor likely limiting the spread of these viruses in the U.S. pig population.IMPORTANCEInterspecies human-to-swine IAV transmission occurs globally and contributes to increased IAV diversity in pig populations. We present data that a swine isolate from a 2018-2019 human-to-swine transmission event was shed for multiple days in challenged and contact pigs. By characterizing this introduction through bioinformatic, molecular, and animal experimental approaches, these findings better inform animal health practices and vaccine decision-making. Since wholly human seasonal H3N2 viruses in the United States were not previously identified as being transmissible in pigs (i.e., reverse zoonosis), these findings reveal that the interspecies barriers for transmission to pigs may not require significant changes to all human seasonal H3N2, although additional changes may be required for sustained transmission in swine populations.

H1N1 swine influenza viruses upregulate NEU1 expression through histone H3 acetylation regulated by HDAC2

H1N1 swine influenza viruses upregulate NEU1 expression through histone H3 acetylation regulated by HDAC2

Influenza A Viruses in the Swine Population: Ecology and Geographical Distribution

Despite the efforts of practical medicine and virology, influenza viruses remain the most important pathogens affecting human and animal health. Swine are exposed to infection with all types of influenza A, B, C, and D viruses. Influenza viruses have low pathogenicity for swine, but in the case of co-infection with other pathogens, the outcome can be much more serious, even fatal. Having a high zoonotic potential, swine play an important role in the ecology and spread of influenza to humans. In this study, we review the state of the scientific literature on the zoonotic spread of swine influenza A viruses among humans, their circulation in swine populations worldwide, reverse zoonosis from humans to swine, and their role in interspecies transmission. The analysis covers a long period to trace the ecology and evolutionary history of influenza A viruses in swine. The following databases were used to search the literature: Scopus, Web of Science, Google Scholar, and PubMed. In this review, 314 papers are considered: n = 107 from Asia, n = 93 from the U.S., n = 86 from Europe, n = 20 from Africa, and n = 8 from Australia. According to the date of publication, they are conditionally divided into three groups: contemporary, released from 2011 to the present (n = 121); 2000–2010 (n = 108); and 1919–1999 (n = 85).

Developing Correlates of Protection for Vaccines Is Needed More than Ever—Influenza, COVID-19 and RSV Infection

One of the greatest success stories of modern medicine is the prevention of infectious diseases by vaccination, most notably against smallpox and poliomyelitis. However, recent events, such as the 2009–2010 swine flu and the 2020 COVID-19 pandemics, as well as the continued emergence of highly pathogenic avian influenza viruses highlighted the fact that we still need to develop new vaccines, and perhaps we should be proactive, rather than reacting to epidemics and pandemics. However, the development of tools for evaluating novel vaccines has not been able to keep up with the rate of vaccine production. Humoral and cellular immune responses to vaccination have both been suggested to be important in preventing infections or ameliorating their consequences, although there is uncertainty regarding their exact roles and importance. This, together with the rapid development of new vaccines, means that the need for developing immunogenicity parameters, and even more importantly, reliable correlates of protection, is more important than ever.

Exploring the Effect of Active Components in Oil Tree Peony Seed Meal on Swine Disease Resistance and its Potential Mechanisms Based on Network Pharmacology and Molecular Docking.

This study aims to explore the feasibility of using network pharmacology and molecular docking technology to predict the effects of active components from oil tree peony seed meal (PSM) on swine diseases. Ten active components of PSM were screened Screening through literature search and network pharmacology standards, including Betulinic acid, Quercetin, Kaempferol, Luteolin, Isorhamnetin, Hydroxygenkwanin, Hederagenin, Benzoyl Paeoniflorin, Albiflorin, Paeoniflorin. Ten types of swine diseases were selected, including African Swine Fever, Aftosa, Swine Vesicular Disease, Transmissible Gastroenteritis, Swine Streptococcal Infection, Blue Aural Disease, Swine Infectious Atrophic Rhinitis, Swine Influenza, Swine Erysipelas, Swine Epidemic Encephalitis. The results showed that the average number of cross genes between the potential target genes of PSM active components and each swine disease target gene accounted for 7.64 % of the total number of swine disease target genes. The GO enrichment analyses showed that putative targets exist in endosomes, lysosomes, cell membranes, nerves, growth factor activity, receptor tyrosine kinase binding, enzyme binding, growth factor binding, transcription coactivator binding, oxidoreductase activity, prostaglandin E receptor activity and insulin receptor substrate binding. The KEGG enrichment analysis results showed that these putative genes were involved in various cancer progression pathways, signaling pathways, and hormone regulatory pathways. A total of 8 core targets were obtained through protein-protein interaction networks analysis, including Protein Kinase CAMP-Activated Catalytic Subunit Alpha (PRKACA), Non-Receptor Tyrosine Kinase (SRC), Mitogen-Activated Protein Kinase 1 (MAPK1), E1A Binding Protein P300 (EP300), Hypoxia Inducible Factor 1 Subunit Alpha (HIF1A), Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta (PIK3CB), C-X-C chemokine receptor type 4 (CXCR4) and Estrogen Receptor 2 (ESR2). The HIF-1 signaling pathway was found to be associated with all 10 selected swine diseases. The PD-L1 expression, and PD-1 checkpoint pathway in cancer, and thyroid hormone signaling pathway were not only enriches the core target with a quantity of 7, but also associated with 9 Swine diseases. In addition, the molecular docking results indicate that the core ingredients have strong affinity with hub genes. The research suggests that the active components of PSM may intervene in swine diseases through multiple components, targets, and pathways.

Two amino acid residues in the N-terminal region of the polymerase acidic protein determine the virulence of Eurasian avian-like H1N1 swine influenza viruses in mice.

Reassortant Eurasian avian-like H1N1 (rEA H1N1) viruses carrying the internal genes of H1N1/2009 virus have been circulating in pigs for more than 10 years and have caused sporadic human infections. The enhanced virulence phenotype of the rEA H1N1 viruses highlights potential risks to public health. However, the molecular mechanism underlying the viral pathogenicity of the currently circulating rEA H1N1 viruses remains unclear. In this study, we found that two naturally isolated rEA H1N1 swine influenza viruses, A/swine/Liaoning/FX38/2017 (FX38) and A/swine/Liaoning/SY72/2018 (SY72), possessed similar genetic characteristics but exhibited significantly different pathogenicity in a mouse model. Using reverse genetics, we demonstrated that amino acid mutations at positions 100 and 122 in the polymerase acidic (PA) protein had individual and synergistic effects on the polymerase activity and viral replication capacity in vitro, as well as the viral pathogenicity in mice. Furthermore, we revealed that amino acid residue 100 in PA influenced the transcription of viral RNA (vRNA) by altering the endonuclease activity, and amino acid residue 122 affected the synthesis of complementary RNA and messenger RNA by altering the RNA-binding ability and endonuclease activity of the PA protein. Taken together, we identified that two naturally occurring amino acid mutations in PA derived from H1N1/2009 virus are crucial determinants of the virulence of rEA H1N1 viruses and revealed the differential mechanism by which these two mutations affect the transcription and replication of vRNA. These findings will extend our understanding of the roles of PA in the virulence of influenza A viruses.IMPORTANCEMultiple genetic determinants are involved in the virulence of influenza A viruses. In this study, we identified two naturally occurring amino acid mutations, located at residues 100 and 122 in the polymerase acidic (PA) protein, which are associated with viral polymerase activity, replication competence, and pathogenicity in mice. In particular, we clarified the specific mechanism by which the two residues play an important role in viral transcription and replication. These findings will help to improve understanding the functions of amino acid residues in the N-terminal region of the PA protein involved in the pathogenicity of influenza A viruses.

Knowledge and Awareness on Swine Origin Influenza A(H1N1) Among Health Care Providers and Civil Employees in a Tertiary Level Hospital, Dhaka Bangladesh

Background: Swine Origin Influenza A(H1N1) viruses create yearly epidemics and rarely pandemics that cost millions of lives worldwide. Health care providers and civil employees at hospital can help in prevention of transmission of influenza by acquiring knowledge and awareness. Methods: A descriptive cross-sectional study was conducted at Combined Military Hospital Dhaka from July to December 2023. Total 213 participants were interviewed that comprises doctors, nurses, medical assistants and civil employees. Purposive sampling technique was used to collect data. Face to face interview was done through a pre-tested semi-structured questionnaire. Data were analysed by using Statistical Package for the Social Sciences (SPSS) version 25.0. Informed consent were obtained from the respondents addressing proper ethical issues. Results: In this study out of total participants 70.4% had previously heard about H1N1 influenza A. Total 38% participants said that internet was found as major source of information. Among the participants 64.3% told fever was common symptom and use of facemask at duty hour was consented by 33.8% as an important means for prevention. Maximum (66.3%) civil employees do not know about washing hand after contact as a preventive procedure leading to spread of infection. Maintaining distance as preventive measures observed by 13.6% participants. Moreover, all participants were aware of transmission through droplet infection except civil employees. In the present study, 50% doctors and 38.1% nurses had knowledge that drug Oseltamivir is very effective against swine Influenza A(H1N1). Remarkable differences were identified among doctors, nurses, medical assistants and civil employees regarding knowledge of quarantine and availability of vaccine. RT-PCR laboratory test is the confirmatory method to defect swine influenza A(H1N1) were exactly known by 9.9% participants, while 70.9% participants don't know the confirm diagnostic method. Conclusion: Knowledge and awareness regarding swine influenza A(H1N1) was low among study participants. For controlling epidemic or even pandemic, awareness by mass media is important. Remarkable gap observed regarding knowledge and awareness about swine influenza A(H1N1) among the health care providers and civil employees, needs to be addressed by proper training and motivation. Bangladesh Armed Forces Med J Vol 57 No (1) June 2024, pp 1-7

Detection of viral agents associated with Porcine Respiratory Disease Complex (PRDC) in pigs in North Kerala

In pigs, stress factors such as pathogenic infections, environmental conditions, and various managemental practices can lead to a disease condition known as Porcine respiratory disease complex (PRDC). The main viral agents associated with this condition are Porcine circovirus 2 (PCV2), Porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine respiratory coronavirus (PRCV), and pseudorabies virus. A study was undertaken to assess whether PCV2 and PRRSV, alone or in combination, are present in cases of respiratory ailments of pigs in North Kerala. The presence of PCV2 was detected by polymerase chain reaction (PCR) targeting the open reading frame 2 (ORF2) of the virus and PRRSV was detected by reverse transcriptase PCR (RT-PCR) targeting ORF6. A total of 54 tissue and lung samples were collected and screened for the presence of PCV2 and PRRSV. Of the samples tested, PCV2 could be detected in 22 (40.74 %) and PRRSV could be detected in 13 (24.07 %) samples. Two tissue samples showed mixed infections with PCV2 and PRRSV. The result shows that viral agents associated with PRDC are prevalent in pigs in North Kerala and that coinfections are also present. Keywords: Porcine respiratory disease complex, Porcine circovirus 2, Porcine reproductive and respiratory syndrome virus, polymerase chain reaction

Potent Anti-Influenza Synergistic Activity of Theobromine and Arainosine.

Influenza represents one of the biggest health threats facing humanity. Seasonal epidemics can transition to global pandemics, with cross-species infection presenting a continuous challenge. Although vaccines and several anti-viral options are available, constant genetic drifts and shifts vitiate any of the aforementioned prevention and treatment options. Therefore, we describe an approach targeted at the virus's channel to derive new anti-viral options. Specifically, Influenza A's M2 protein is a well-characterized channel targeted for a long time by aminoadamantane blockers. However, widespread mutations in the protein render the drugs ineffective. Consequently, we started by screening a repurposed drug library against aminoadamantane-sensitive and resistant M2 channels using bacteria-based genetic assays. Subsequent in cellulo testing and structure-activity relationship studies yielded a combination of Theobromine and Arainosine, which exhibits stark anti-viral activity by inhibiting the virus's channel. The drug duo was potent against H1N1 pandemic swine flu, H5N1 pandemic avian flu, aminoadamantane-resistant and sensitive strains alike, exhibiting activity that surpassed Oseltamivir, the leading anti-flu drug on the market. When this drug duo was tested in an animal model, it once more outperformed Oseltamivir, considerably reducing disease symptoms and viral RNA progeny. In conclusion, the outcome of this study represents a new potential treatment option for influenza alongside an approach that is sufficiently general and readily applicable to other viral targets.

Antiviral Susceptibility of Swine-Origin Influenza A Viruses Isolated from Humans, United States.

Since 2013, a total of 167 human infections with swine-origin (variant) influenza A viruses of A(H1N1)v, A(H1N2)v, and A(H3N2)v subtypes have been reported in the United States. Analysis of 147 genome sequences revealed that nearly all had S31N substitution, an M2 channel blocker-resistance marker, whereas neuraminidase inhibitor-resistance markers were not found. Two viruses had a polymerase acidic substitution (I38M or E199G) associated with decreased susceptibility to baloxavir, an inhibitor of viral cap-dependent endonuclease (CEN). Using phenotypic assays, we established subtype-specific susceptibility baselines for neuraminidase and CEN inhibitors. When compared with either baseline or CEN-sequence-matched controls, only the I38M substitution decreased baloxavir susceptibility, by 27-fold. Human monoclonal antibodies FI6v3 and CR9114 targeting the hemagglutinin's stem showed variable (0.03 to >10 µg/mL) neutralizing activity toward variant viruses, even within the same clade. Methodology and interpretation of laboratory data described in this study provide information for risk assessment and decision-making on therapeutic control measures.

A Narrative Review on the Pandemic Zoonotic RNA Virus Infections Occurred During the Last 25 Years.

Pandemic zoonotic RNA virus infections have continued to threaten humans and animals worldwide. The objective of this review was to highlight the epidemiology and socioeconomic impacts of pandemic zoonotic RNA virus infections that occurred between 1997 and 2021. Literature search was done from Web of Science, PubMed, Google Scholar and Scopus databases, cumulative case fatalities of individual viral infection calculated, and geographic coverage of the pandemics were shown by maps. Seven major pandemic zoonotic RNA virus infections occurred from 1997 to 2021 and were presented in three groups: The first group consists of highly pathogenic avian influenza (HPAI-H5N1) and swine-origin influenza (H1N1) viruses with cumulative fatality rates of 53.5% and 0.5% in humans, respectively. Moreover, HPAI-H5N1 infection caused 90-100% death in poultry and economic losses of >$10billion worldwide. Similarly, H1N1 caused a serious infection in swine and economic losses of 0.5-1.5% of the Gross Domestic Product (GDP) of the affected countries. The second group consists of severe acute respiratory syndrome-associated coronavirus infection (SARS-CoV), Middle East Respiratory Syndrome (MERS-CoV) and Coronavirus disease 2019 (COVID-19) with case fatalities of 9.6%, 34.3% and 2.0%, respectively in humans; but this group only caused mild infections in animals. The third group consists of Ebola and Zika virus infections with case fatalities of 39.5% and 0.02%, respectively in humans but causing only mild infections in animals. Similar infections are expected in the near future, and hence strict implementation of conventional biosecurity-based measures and development of efficacious vaccines would help minimize the impacts of the next pandemic infection.

Monitoring of Respiratory Disease Patterns in a Multimicrobially Infected Pig Population Using Artificial Intelligence and Aggregate Samples

A 24/7 AI sound-based coughing monitoring system was applied in combination with oral fluids (OFs) and bioaerosol (AS)-based screening for respiratory pathogens in a conventional pig nursery. The objective was to assess the additional value of the AI to identify disease patterns in association with molecular diagnostics to gain information on the etiology of respiratory distress in a multimicrobially infected pig population. Respiratory distress was measured 24/7 by the AI and compared to human observations. Screening for swine influenza A virus (swIAV), porcine reproductive and respiratory disease virus (PRRSV), Mycoplasma (M.) hyopneumoniae, Actinobacillus (A.) pleuropneumoniae, and porcine circovirus 2 (PCV2) was conducted using qPCR. Except for M. hyopneumoniae, all of the investigated pathogens were detected within the study period. High swIAV-RNA loads in OFs and AS were significantly associated with a decrease in respiratory health, expressed by a respiratory health score calculated by the AI The odds of detecting PRRSV or A. pleuropneumoniae were significantly higher for OFs compared to AS. qPCR examinations of OFs revealed significantly lower Ct-values for swIAV and A. pleuropneumoniae compared to AS. In addition to acting as an early warning system, AI gained respiratory health data combined with laboratory diagnostics, can indicate the etiology of respiratory distress.

Reverse zoonotic transmission of human seasonal influenza to a pig herd in Sweden.

In January 2023, a Swedish piglet-producing farm with 2800 sows in production (SIP) was diagnosed with IAV (Influenza A virus) and the isolates were shown to cluster with the human seasonal influenza (2022/2023). In December 2022, employees with flu like symptoms tended to the pigs and a few weeks later, respiratory signs appeared in different age groups; sows in farrowing units were anorectic and pyrectic. Lung and nasal swabs were tested positive for IAV and other respiratory infectious agents. Blanket vaccination against H1N1pdm09 of sows and gilts was initiated but discontinued for sows after 2 treatments. Biosecurity measures aiming to reduce the spread of virus were implemented. However, the compliance to follow the protocol was moderate.Combining immunity and strict sanitary measures is crucial to control virus circulation. As the farmer discontinued sow vaccination and just partly increased biosecurity, this may have contributed to ongoing virus circulation and clinical signs in pigs, even 5 months post-diagnosis. Although H1N1pdm09 already had been found in the herd in 2017, there were no clinical signs or diagnostic results indicating continuous circulation of this or other IAV strains afterwards. However, this cannot be entirely excluded. Swine IAVs pose a risk of reintroduction into the human population, highlighting the importance of vaccination of farm workers against seasonal influenza.

Exploring the Genetic Diversity of Mycoplasma hyopneumoniae in Pigs with Pneumonia and Pleurisy at Slaughter.

Mycoplasma (M.) hyopneumoniae is the key pathogen of the porcine respiratory disease complex (PRDC) and contributes to pleurisy in pigs. Due to its limited metabolism and laborious cultivation, molecular tools are useful for diagnosis. This study investigated the genetic diversity of M. hyopneumoniae in slaughter pigs with pneumonia and pleurisy, and it assessed co-infections by Pasteurella multocida type A (PM), Actinobacillus pleuropneumoniae (APP), and swine influenza virus A (sIVA). Lungs (n = 70) with different pleurisy scores and lesions compatible with M. hyopneumoniae infection were collected for convenience. Macroscopic and microscopic evaluations were performed. M. hyopneumoniae was detected using qPCR, and MLST was used for genetic characterization. Co-infections with PM and APP were also evaluated by qPCR, while the immunohistochemistry assessed sIVA infection. All lungs were positive for M. hyopneumoniae. Histopathology confirmed M. hyopneumoniae-associated lesions. MLST characterization was possible in 25 lungs and revealed 10 distinct allelic profiles, with none matching known sequence types in the public database. Co-infections were detected in 40% of the samples with APP and 32% with PM, with 12% showing both pathogens and 52% of the samples presenting microscopic lesions compatible with sIVA infection. The diverse genetic profiles found underscore the need for research on isolation and potential pathogenic variations.

Discriminating North American Swine Influenza Viruses with a Portable, One-Step, Triplex Real-Time RT-PCR Assay, and Portable Sequencing.

Swine harbors a genetically diverse population of swine influenza A viruses (IAV-S), with demonstrated potential to transmit to the human population, causing outbreaks and pandemics. Here, we describe the development of a one-step, triplex real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay that detects and distinguishes the majority of the antigenically distinct influenza A virus hemagglutinin (HA) clades currently circulating in North American swine, including the IAV-S H1 1A.1 (α), 1A.2 (β), 1A.3 (γ), 1B.2.2 (δ1) and 1B.2.1 (δ2) clades, and the IAV-S H3 2010.1 clade. We performed an in-field test at an exhibition swine show using in-field viral concentration and RNA extraction methodologies and a portable real-time PCR instrument, and rapidly identified three distinct IAV-S clades circulating within the N.A. swine population. Portable sequencing is used to further confirm the results of the in-field test of the swine triplex assay. The IAV-S triplex rRT-PCR assay can be easily transported and used in-field to characterize circulating IAV-S clades in North America, allowing for surveillance and early detection of North American IAV-S with human outbreak and pandemic potential.

A Study to Assess the Effectiveness of Structured Teaching Program on Knowledge Regarding Prevention of Swine Flu Among in Rural Area in Indore City

Pigs around the world are susceptible to swine influenza, a respiratory disease brought on by type A influenza viruses, particularly strains H1N1, H1N2, H2N1, H3N1, H3N2, and H2N3. Swine influenza viruses can infect humans and cause mild to severe sickness in people of all ages. Those who come into contact with pigs are particularly at risk. Influenza epidemiology research was previously only done in resource-rich nations. After 30,000 cases of H1N1 illness were recorded from more than 70 countries on June 11, 2009, the World Health Organization proclaimed an H1N1 pandemic. The frequency of swine influenza significantly increased in 2015, reaching a 5-year high. 2015 saw 10,000 instances of swine flu confirmed in India, with 774 fatalities. KEYWORDS : Prevention from Swine flu, Rural area, Structured Teaching Programme

COBRA N2 NA vaccines induce protective immune responses against influenza viral infection

ABSTRACT Influenza neuraminidase (NA) is a promising target for a broadly protective vaccine. In this study, the Computationally Optimized Broadly Reactive Antigen (COBRA) methodology was used to develop N2 NA vaccine candidates. The unique wild type (WT) N2 sequences of human and swine influenza strains isolated between 1957 and 2019 were used to design the COBRA N2-A NA vaccine, while the unique WT N2 sequences of human influenza strains isolated between 2000 and 2019 were used to design the COBRA N2-B NA vaccine. Sera collected from COBRA N2 NA vaccinated mice showed more broadly reactive antibody responses against a broad panel of H×N2 influenza virus strains than sera collected from mice vaccinated with WT N2 NA vaccines. Antibodies elicited by COBRA or WT N2 NA antigens cross react with recent human H3N2 influenza viruses from different clades, while the antibodies elicited by A/Switzerland/9715293/2013 hemagglutinin (HA) reacted with viruses from the same clade. Furthermore, mice vaccinated with COBRA N2-B NA vaccine had lower viral lung titers compared to mock vaccinated mice when challenged with human H3N2 influenza viruses. Thus, the COBRA N2 NA vaccines elicit broadly protective murine anti-NA antibodies against multiple strains across subtypes and the viral loads were significantly decreased in the lungs of the mice in the COBRA N2 NA vaccine groups, compared to the mice in the mock vaccinated group, indicating that the COBRA-based N2 subtype NA vaccines have a potential to be a component in a universal influenza vaccine.