Virus Subtype Research Articles

A human isolate of bovine H5N1 is transmissible and lethal in animal models.

The outbreak of clade 2.3.4.4b highly pathogenic avian influenza viruses of the H5N1 subtype (HPAI H5N1) in dairy cows in the US has so far resulted in spillover infections of at least thirteen farm workers1-3, who presented with mild respiratory symptoms or conjunctivitis, and one individual with no known animal exposure who was hospitalized but recovered3,4. Here, we characterized A/Texas/37/2024 (huTX37-H5N1), a virus isolated from the eyes of an infected farm worker who developed conjunctivitis5. huTX37-H5N1 replicated efficiently in primary human alveolar epithelial cells, but less efficiently in corneal epithelial cells. Despite causing mild disease in the infected worker, huTX37-H5N1 was lethal in mice and ferrets and spread systemically with high titres in respiratory and non-respiratory organs. Importantly, in four independent experiments in ferrets, huTX37-H5N1 transmitted via respiratory droplets in 17%-33% of transmission pairs and five of six exposed ferrets that became infected died. PB2-631L (encoded by bovine isolates), promoted influenza polymerase activity in human cells, suggesting a role in mammalian adaptation like that of PB2-627K (encoded by huTX37-H5N1). Additionally, bovine HPAI H5N1 viruses were found to be susceptible to polymerase inhibitors both in vitro and in mice. Thus, HPAI H5N1 virus derived from dairy cattle transmits by respiratory droplets in mammals without prior adaptation and causes lethal disease in animal models.

Application of surface plasmon resonance imaging in the high-throughput detection of influenza virus.

To evaluate the application effect of SPRi monoclonal antibody (mAb) chip in the detection of influenza virus antigen in complex mixtures. A total of 115 strains of mAbs against different subtypes (H1N1, H5N1, A1, A3, B, H7N9, H9N2, and H3N2) of influenza virus were prepared. The chip of mAbs against influenza virus was prepared by surface plasmonic resonance imaging (SPRi) technology, which was used for the detection of influenza virus supernatant, and compared with the traditional antigen capture ELISA method. Comparative studies have shown that traditional antigen capture ELISA methods have a higher sensitivity (86.8% (46/53) vs. 46.5% (46/99); z = 4.84, P < 0.001), while the SPRi chip methods present a significantly higher specificity (56.3% (9/16) vs. 14.5% (9/62); z = 3.54, P < 0.001). The SPRi chip detection method for influenza virus antibodies can well reflect the specific binding characteristics of influenza virus antigens and antibodies. The SPRi mAb chip can be used for the detection of specific pathogenic microorganisms or viral proteins in complex mixtures such as influenza virus supernatant. It has significant advantages of label free, real-time, high-throughput, and good specificity, and can play an important role in disease diagnosis and infectious disease prevention and control.

Influenza Virus Types, Subtypes and Genomic Lineage with Its Prevention and Control: A Narrative Review

Since influenza viruses continue to be a serious hazard to both humans and animals, they are still very important. Types A, B, and C influenza viruses are among the members of the Orthomyxoviridae family. A quick summary of the molecular factors is provided, which determine pathogenicity and clinical signs and symptoms of influenza. A review of host range and evolution highlights the genetic diversity of influenza A viruses and their capacity to successfully infect a variety of hosts, including avian and mammalian species. Moreover, influenza viruses may reassemble segments because of the way their segmented genome is designed. The significance of host sialic acid distribution and viral receptor-binding hemagglutinins in species-restricted virus binding is emphasized. It results in yearly outbreaks and necessitates the creation of novel vaccination formulations. This may ultimately result in the creation of a virus that can spread among humans and has unique antigenic qualities, perhaps sparking a pandemic. Current developments in our knowledge of the seasonality, transmission, and prevention of influenza viruses are outlined, along with their significance for halting the virus's spread. Journal of Monno Medical College June, 2024; 10 (1):43-51

Long-Lasting Chemiluminescence Based on Functionalized Multicolor Protein Capsules for Multiple Visualization Detection of Avian Influenza Virus Biomarkers.

Long-lasting chemiluminescence (CL) emissions are necessary for improving the detection accuracy and expanding the application scope. Here, we have synthesized three oil-in-water (O/W) multicolor protein capsules (LCBA, F/LCBA, and RB/F/LCBA) using a simple ultrasound method and have engineered specific target-triggered catalytic hairpin assembly on their surface and chemiluminescence resonance energy transfer inside. Consequently, three multicolor capsules exhibit excellent structural stability, generate blue-, green-, and red-colored emissions when reacting with H2O2, have long-lasting CL emission over 1 h, and successfully achieve the accurate multiple visualization detection of avian influenza virus subtype targets. Without the need for complex instruments and analysis procedures, the CL imaging assays can be carried out and recorded with a common smartphone. The detection limits for visualizing H1N1, H7N9, and H5N1 are 5.5, 7.6, and 9.0 pM, respectively. There is a linear range between 20.0 and 625 pM and excellent selectivity against interfering DNA. Furthermore, visualization detection has been successfully applied for the detection of H1N1, H7N9, and H5N1 in healthy human serum samples. With these merits, this facile, ultrasensitive, and multiple visualization sensor has potential applications in point-of-care testing and early diagnosis.

Inserting CTL Epitopes of the Viral Nucleoprotein to Improve Immunogenicity and Protective Efficacy of Recombinant Protein against Influenza A Virus.

Conserved influenza virus proteins, such as the hemagglutinin stem domain (HA2), nucleoprotein (NP), and matrix protein (M), are the main targets in the development of universal influenza vaccines. Previously, we constructed a recombinant vaccine protein Flg-HA2-2-4M2ehs containing the extracellular domain of the M2 protein (M2e) and the aa76-130 sequence of the second HA subunit as target antigens. It demonstrated immunogenicity and broad protection against influenza A viruses after intranasal and parenteral administration. This study shows that CD8+ epitopes of NP, inserted into a flagellin-fused protein carrying M2e and HA2, affect the post-vaccination immune humoral response to virus antigens without reducing protection. No differences were found between the two proteins in their ability to stimulate the formation of follicular Th in the spleen, which may contribute to a long-lasting antigen-specific humoral response. The data obtained on Balb/c mice suggest that the insertion of CTL NP epitopes into the flagellin-fused protein carrying M2e and HA2 reduces the antibody response to M2e and A/H3N2. In C57Bl6 mice, this stimulates the formation of NP-specific CD8+ Tem and virus-specific mono- and multifunctional CD4+ and CD8+ Tem in the spleen and completely protects mice from influenza virus subtypes A/H1N1pdm09 and A/H3N2.

Pathogenetic investigation of an outbreak of upper respiratory tract infection in a kindergarten in Baiyin City, Gansu Province

This study focuses on the cases(mainly characterized by respiratory symptoms such as cough, runny nose, fever, sore throat, and nasal congestion)of an outbreak of upper respiratory tract infections in a kindergarten in Jingyuan County, Baiyin City, Gansu Province, in May 2023. The epidemiological data were collected, and pharyngeal swab specimens were also obtained from the patients. The specimens of the research participants were subjected to respiratory multi-pathogen testing, and the positive specimens were further analyzed by sequencing the second hypervariable region (HRV2) of the G gene of respiratory syncytial virus (RSV) and constructing a phylogenetic tree. A total of 90 patients were collected, with an incidence rate of 22.84% (90/394), and the highest incidence was observed in the junior class group at 29.55%. Among the 17 pharyngeal swab specimens collected, 16 specimens were identified with the A subtype of respiratory syncytial virus. Sequencing analysis confirmed that it was the A subtype ON1 genotype. Based on the aforementioned testing results, it can be concluded that the current epidemic was primarily caused by infection with the A subtype of respiratory syncytial virus. Following the implementation of intervention measures, the epidemic has been effectively controlled.

Diversity, Geographical Distribution and Predictive Factors of Hepatitis C Virus Genotypes and Subtypes in Rwanda

Existing data on the prevalence of hepatitis C virus (HCV) genotypes and subtypes in Rwanda need to be strengthened. The aim of this study was to identify HCV genotypes and subtypes among HCV-infected patients, as well as their geographical distribution in Rwanda, and to identify the social and economic factors that could influence HCV epidemiology which would make it possible to target national preventive and management actions for infected patients. This study included 560 patients with confirmed chronic HCV infection. Patients were recruited from various health facilities in the four provinces of Rwanda as well as in the City of Kigali and had never received treatment with direct-acting antiviral (DAAs). HCV viral loads were measured using Cobas® AmpliPrep/Cobas® TaqMan® HCV Quantitative Test, version 2.0. HCV genotyping was performed using an in-house sequencing protocol targeting the NS5B central region. Genotypic HCV prevalence was correlated with patient geographic location, sociodemographic, behavioral, lifestyle, and clinical factors. HCV genotype 4 was detected in 99.3% of the patients, while genotype 3 was identified in 0.7%. A total of eight (8) HCV subtypes were detected, with 4k being the predominant subtype nationwide (49.5%), followed by subtypes 4r (21.2%), 4q (16.2%), 4v (7.9%), 4b (2.0%), 4l (1.8%), 4c and 3h represent 0.7% each. Our findings reveal subtype distribution variations among provinces. Subtype 4k was prevalent across regions, particularly in Kigali (64.0%) and the Eastern Province (61.6%). Subtype 4q was more common in the northern province (40.7%), 4r in the southern (43.9%) and western provinces (37.1%), and 4v in the eastern province (17.8%). Farmers exhibit a distinct infection profile compared to other occupations, showing a lower prevalence of subtype 4k but a higher prevalence of subtype 4r. Our study revealed that HCV infection is unevenly distributed in Rwanda, dominated by HCV genotype 4, with considerable heterogeneity in the repartition of the different subtypes. We found potential associations between rural/urban lifestyles and HCV subtype profiles. Determined HCV distribution and diversity can serve as basis not only for HCV infection awareness and prevention campaigns, but also success and guidance for personalized treatment.

Circulating respiratory viruses including SARS-CoV-2 during 2021-2022 season in Tunisia: Epidemiological and dynamic changes

BackgroundChanging patterns in community respiratory virus activity were reported in different geographical locations during the COVID-19 pandemic. In this study, we aimed to assess the prevalence of circulating respiratory viruses, including SARS-CoV-2, during the season 2021-2022 in Tunisia. MethodsWe retrospectively enrolled 328 nasopharyngeal samples received at the Triage Center of Habib Bourguiba Hospital from patients with acute respiratory symptoms during September 2021-May 2022. All samples were screened for both SARS-CoV-2 and common respiratory viruses. This latter detection was performed using end-point multiplex RT-PCRs, Real-Time PCR, and AllplexTM Respiratory Panel 1 kit (Seegene) for Influenza Virus A (IFVA) and Respiratory Syncytial Virus (RSV) subtyping. ResultsAmong included patients, at least one viral pathogen was identified in 118 (35.9%) patients. The detection rate of SARS-CoV-2 was 21.6%. A low viral coinfection rate was observed (3.3%). The most prevalent pathogen among non-SARS-CoV-2 viruses was Enterovirus/Rhinovirus (HEV/HRV) (59.6%) followed by IFVA (15.3%) and Adenoviruses (ADV) (11.5%). Only IFVA H3N2 was found to circulate during the study period. A negative virus interaction was eventually induced by SARS-CoV-2, as it was shown by lower levels of activity of non-SARS-CoV-2 viruses (not exceeding 17.7%) while infections due to pandemic Omicron variants of concern became widespread. ConclusionsThis study highlights the relative return of community IFVA circulation during the 2021-2022 season in Tunisia. A negative viral interaction between SARS-CoV-2 and other respiratory viruses is highly suggested, which explains, in addition to the easing of COVID-19 restriction measures, the epidemiological changes in non-SARS-CoV-2 viruses circulation.

Application of lentinan in suppression of Marek's disease virus infection

Marek's disease virus (MDV) is an extremely widespread avian immunosuppressive virus. In recent years, many reports have shown that there are still cases of MDV infection and immunosuppression after immunization with the vaccine. Consequently, there is a need to develop alternative complementary approaches for strengthening the efficacy of MDV prevention and control measures. Lentinan (LNT) is a macromolecular compound with immune-enhancing activity extracted from shiitake mushrooms. To explore the value and effectiveness of administering LNT through drinking water in the prevention and control of MDV, this study first observed the effects of high and low doses of LNT on weight gain, organ development, viral replication, and antibody titer of an avian influenza virus subtype H9 (AIV-H9) inactivated vaccine in specific pathogen-free (SPF) chicks infected with the wild strain of MDV. The results showed that both high and low doses of LNT significantly alleviated the weight gain retardation and liver and spleen enlargement caused by MDV infection, and significantly inhibited the replication of MDV in SPF chicks (P < 0.05), compared with the MDV-positive control group, both high and low doses of LNT significantly increased the antibody titer of AIV-H9 after immunization with inactivated AIV-H9 vaccine (P < 0.0001). On this basis, we also observed the effects of a chicken Marek's disease meq gene deletion live vaccine (SC9-1 strain), administered alone or in combination with LNT, on MDV infections of varying virulence in Hy-Line Brown chicks. The results showed that combined administration of LNT and the SC9-1 vaccine resulted in a significant alleviation of weight gain retardation and liver and spleen enlargement due to MDV infection (P < 0.05), as well as a significant inhibition of MDV replication and release in Hy-Line Brown chicks compared to the vaccine alone (P < 0.0001). These findings suggest that LNT not only alleviates the adverse effects of MDV infection in chickens but also enhances the efficacy of MDV vaccination, offering a potential auxiliary measure for controlling MDV infection.

Pichia pastoris an Ideal Host for the Production of Recombinant Influenza Vaccines

Pichia pastoris is a methylotrophic yeast with remarkable characteristics such as lacking endotoxin, producing high amounts of recombinant protein, performing post-translational modifications, and so on. Influenza A virus, a member of the Orthomyxoviridae family, is the cause of avian influenza. Three avian influenza virus subtypes, H5, H7 and H9, are commercially and physiologically significant in the poultry industry. Some researchers considered influenza to be the next pandemic disease. Nowadays, researchers have paid attention to producing novel and effective recombinant vaccines, especially in the poultry industry. Due to the advantages of P. pastoris yeast, it can be used as an ideal expression system for producing subunit vaccines. Although several studies have been conducted in this field, there is no comprehensive review of using P. pastoris to produce recombinant influenza vaccines. This review explains the different strains, phenotypes, and advantages of this yeast and then the production of recombinant influenza vaccines using this expression system is discussed in detail.

Experience with HCV Detection and Molecular Genetic Characterization among Otherwise Healthy Pregnant Women and Their Partners in the Republic of Guinea.

According to recent data, there are currently 170 to 200 million people infected with HCV worldwide, and the number of new cases annually is approximately 40,000. Thus, the overall prevalence of the pathogen in the world is about 1.8-3%. The dynamic monitoring of circulating viral variants in specific groups that reflect the situation in the wider population, including potential pathogen spread, is of high importance for predicting the epidemiologic situation. Pregnant women are such a group. The Republic of Guinea is one of the poorest countries in the world, in which medicine receives little finance from the state. Among other conditions, HCV infection is not monitored in the country. This work used blood plasma from pregnant women living in the Republic of Guinea and their partners (1810 and 481). ELISA diagnostic kits were used to detect serologic markers, and PCR diagnostic kits were used to detect molecular biologic markers. Sanger sequencing, followed by phylogenetic analysis, was used for genotyping. The present study shows that HCV antibodies were detected in 3.2% of the pregnant women examined and in 3.33% of their male partners. HCV RNA was detected in 0.5% of cases in women and in all anti-HCV-positive male partners (3.33%). HCV RNA was more common in the men than in the pregnant women (χ2 = 25.6, df 1, p < 0.0001, RR = 6.69 with 95% CI: 2.97-15.04). The HCV viral load was determined for all the RNA-HCV-positive samples. The HCV viral load exceeded 1000 IU/mL in all nine women and only in two cases in men. The HCV genes NS5A and NS5B and the NS3 gene fragment were sequenced for 11 samples. Subtype 2q was determined for three isolates and 2j for another three isolates. Another five isolates could not be confidently assigned a subtype because different results were obtained with different methods of analyzing the three viral regions. It can be assumed that these isolates belong to new viral subtypes or to recombinant forms between genotype 2 subtypes. No drug resistance mutations were identified, but a large number of natural polymorphisms in the analyzed genomic regions of the HCV isolates were shown. These results may serve as baseline data for the future planning of a nationwide estimate of the prevalence of bloodborne infections among pregnant women.

Transmission network of Hepatitis C virus subtype 2a in Huazhou County, Shaanxi Province, China

BackgroundHuazhou County has one of the highest rates of hepatitis C virus (HCV) infection incidence and prevalence in Shaanxi Province, northwest China. Understanding the characteristics of HCV transmission patterns in this area could help guide targeted prevention strategies. This study employed phylogenetic analysis and the construction of a molecular transmission network of HCV-infected people in Huazhou County to describe the predominant strains of HCV and identify factors associated with onward transmission.MethodsWhole blood samples were obtained from HCV RNA-positive individuals for sequencing of the non-structural protein 5B region. A maximum-likelihood (ML) phylogenetic tree was constructed to determine HCV subgenotypes, and Bayesian phylogenetic analysis was employed to estimate the evolutionary history. The transmission network was constructed using the ML phylogenetic tree and pairwise distances. Logistic regression was used to identify factors associated with clustering in the transmission network.ResultsML phylogenetic analysis confirmed that the 61 sequences analyzed in the study belonged to subtype 2a. Bayesian phylogenetic analysis showed that the majority of subtype 2a sequences originated in the northwest of China and had descended approximately 8 to 20 years before sampling. Overall, 26.2% of participant sequences were grouped into phylogenetic network clusters. Multivariate logistic regression showed that individuals who had a history of blood transfusions and were living in Shi Village, Huazhou County, were more likely to form clusters within the transmission network.ConclusionHCV transmission in Huazhou County was predominantly associated with subtype 2a. Having a history of blood transfusions and living in residential Shi Village, Huazhou County, were factors associated with a high risk of HCV infection transmission. Prioritizing targeted interventions for these patient groups may help to prevent further infections.

First detection of influenza A virus subtypes H1N1 and H3N8 in the Antarctic region: King George Island, 2023.

Influenza A virus is characterized by a segmented single-stranded RNA genome. Such organization of the virus genome determines the possibility of reassortment, which can lead to the emergence of new virus variants. The main natural reservoir of most influenza A virus subtypes are wild waterfowl. Seasonal migrations gather waterfowl from all major migration routes to nesting areas near the northern and southern polar circles. This makes intercontinental spread of influenza A viruses possible. Objective ‒ to conduct molecular genetic monitoring and study the phylogenetic relationships of influenza A virus variants circulating in Antarctica in 2023. We studied 84 samples of biological material obtained from birds and marine mammals in April‒May 2023 in coastal areas of Antarctica. For 3 samples, sequencing was performed on the Miseq, Illumina platform and phylogenetic analysis of the obtained nucleotide sequences of the influenza A virus genomes was performed. The circulation of avian influenza virus in the Antarctic region was confirmed. Heterogeneity of the pool of circulating variants of the influenza A virus (H3N8, H1N1) was revealed. Full-length genomes of the avian influenza virus were sequenced and posted in the GISAID database (EPI_ISL_19032103, 19174530, 19174467). The study of the genetic diversity of influenza A viruses circulating in the polar regions of the Earth and the identification of the conditions for the emergence of new genetic variants is a relevant task for the development of measures to prevent biological threats.

Serological monitoring of Influenza A among wild and domestic ungulates in Ukraine

The article provides a brief historical background of equine influenza, the spread of this disease worldwide, and the current epizootic situation. The results of serological monitoring by ELISA of wild and domestic ungulates from different farms and regions of Ukraine for the presence of antibodies to influenza A viruses are presented. Blood serum samples from 372 domestic horses and 32 wild ungulates were tested. Samples from animals collected in 2023 and 2024 and archival blood serum samples from 2021 were used and tested according to the manufacturer’s instructions using ELISA test systems manufactured by IDEXX, INGEZIM, and IDVet. The data obtained indicate a fairly active circulation of influenza A viruses in populations of unvaccinated domestic horses. The circulation was established not only in recent years (2023–2024, seroprevalence from 10% to 100%), but was observed earlier, as evidenced by the detection of 60.9% of positive samples in samples collected in 2021. In addition, two out of three positive samples were found in wild horses from Kherson Region, which indicates the circulation of influenza A virus among wild animals and requires further investigation. The results correlate with the worsening of the epidemiological situation regarding influenza in animals in Europe. The subsequent phase of the research is serotyping, which involves determining the presence of antibodies to specific virus subtypes by hemagglutinin

Wastewater Surveillance for Influenza A Virus and H5 Subtype Concurrent with the Highly Pathogenic Avian Influenza A(H5N1) Virus Outbreak in Cattle and Poultry and Associated Human Cases — United States, May 12–July 13, 2024

Wastewater Surveillance for Influenza A Virus and H5 Subtype Concurrent with the Highly Pathogenic Avian Influenza A(H5N1) Virus Outbreak in Cattle and Poultry and Associated Human Cases — United States, May 12–July 13, 2024

A post-marketing study to evaluate the safety and immunogenicity of a quadrivalent influenza split-virion vaccine in elderly people aged 60 years and older

BackgroundInfluenza remains a global public health concern. Understanding the vaccination-induced response in an aging population, which is susceptible and at high risk, is essential for disease prevention and control. Here, we report findings on the safety and immunogenicity of a quadrivalent influenza split-virion vaccine (15 µg/subtype/0.5 ml/dose) (hereinafter referred to as the “quadrivalent influenza vaccine”) in a population aged ≥ 60 years.MethodsThis open-label, pragmatic post-marketing trial enrolled 1399 older adults to receive one dose of an approved commercially available quadrivalent influenza vaccine manufactured by Hualan Biological Bacterin Inc. (hereinafter referred to as “Hualan Bio”). Participants with contraindications for the vaccine were excluded, while poor health condition was acceptable. All vaccinated subjects experienced adverse events collection within 30 days and serious adverse events within 180 days post-vaccination. 25% subjects, selected randomly, underwent venous blood sampling pre-vaccination and 30 days after post-vaccination, for detecting antibody titers against each subtype of influenza virus by hemagglutination inhibition assay. The incidences of adverse events and antibody titers against each subtype of influenza virus were statistically analyzed using SAS 9.4.ResultsNo grade 3 adverse reactions occurred within 30 days post-vaccination. The incidences of overall adverse reactions, local adverse reactions and systemic adverse reactions were 3.79%, 2.86% and 1.00%, respectively. No serious adverse reactions occurred within 180 days post-vaccination. There were 350 subjects who completed venous blood sampling pre-vaccination, among whom 348 subjects completed venous blood sampling at 30 days post-vaccination for immunogenicity assessment. With respect to hemagglutination inhibition antibodies against influenza viruses H1N1, H3N2, BV and BY subtypes, at 30 days post-vaccination, the seroconversion rates were 87.64%, 75.57%, 73.28% and 78.74%, respectively; the seropositive rates were 93.97%, 98.56%, 79.31% and 95.40%, respectively; and the geometric mean increase (GMI) in post-immunization/pre-immunization antibodies was 24.80, 7.26, 10.39 and 7.39, respectively.ConclusionOne 15 µg/subtype dose of the vaccine had a good safety profile and elicited favorable immunogenicity among subjects aged ≥ 60 years. The results of this study indicate that Hualan Bio quadrivalent influenza vaccine strike balance between safety and immunogenicity, supporting unnecessity to increase dosage or inoculation frequency for further enhancing immunogenicity.Trial registrationRegistered on ClinicalTrials.gov. Registration number: NCT06334510. Registered on 28/03/2024 (retrospectively registered).

Analyzing Molecular Traits of H9N2 Avian Influenza Virus Isolated from a Same Poultry Farm in West Java Province, Indonesia, in 2017 and 2023

Background Indonesia is one of the countries that is endemic to avian influenza virus subtype H9N2. This study aims to compare the molecular characteristics of avian influenza virus (AIV) subtype H9N2 from West Java. Methods Specific pathogen-free (SPF) embryonated chicken eggs were used to inoculate samples. RNA extraction and RT–qPCR confirmed the presence of H9 and N2 genes in the samples. RT–PCR was employed to amplify the H9N2-positive sample. Nucleotide sequences were obtained through Sanger sequencing and analyzed using MEGA 7. Homology comparison and phylogenetic tree analysis, utilizing the neighbor-joining tree method, assessed the recent isolate’s similarity to reference isolates from GenBank. Molecular docking analysis was performed on the HA1 protein of the recent isolate and the A/Layer/Indonesia/WestJava-04/2017 isolate, comparing their interactions with the sialic acids Neu5Ac2-3Gal and Neu5Ac2-6Gal. Results RT–qPCR confirmed the isolate samples as AIV subtype H9N2. The recent virus exhibited 11 amino acid residue differences compared to the A/Layer/Indonesia/WestJava-04/2017 isolate. Phylogenetically, the recent virus remains within the h9.4.2.5 subclade. Notably, at antigenic site II, the recent isolate featured an amino acid N at position 183, unlike A/Layer/Indonesia/WestJava-04/2017. Molecular docking analysis revealed a preference of HA1 from the 2017 virus for Neu5Ac2-3Gal, while the 2023 virus displayed a tendency to predominantly bind with Neu5Ac2-6Gal. Conclusion In summary, the recent isolate displayed multiple mutations and a strong affinity for Neu5Ac2-6Gal, commonly found in mammals.

The dynamics of the cytokine profile of the lungs and liver of BALB/C male mice infected with the H1N1 subtype avian influenza virus

The increasing number of reported cases of direct transmission of avian influenza viral strains to humans is obviously associated with the threat of a new pandemic and the need for a more detailed study of the pathogenesis of the viral infectious process. The aim of the research was to study the pathogenesis of influenza infection and in particular to establish the role of proinflammatory cytokines in the pathogenesis of the inectious process caused by influenza virus subtype A/WSN/1/33(H1N1). The experiment involved 21 male BALB/c mice aged 4-6 weeks weighing 16-18 g, which16were infected intranasally with influenza A/WSN/1/33(H1N1) virus under ether anesthesia by inoculation of 50 µl of allantoic fluid containing 10 LD50 of the virus. Histological studies were performed on samples of lung and liver tissues. It was evidenced that infection caused by the highly pathogenic for mice influenza virus A/WSN/1/33(H1N1) initiates the spreading of dystrophic and necrotic processes mainly in the lungs and liver. This can occur due the increase of proinflammatory cytokines leading to the violation of the circulatory system triggered by the active reproduction of the virus and toxic manifestations on behalf of cellular immunity. Structural changes in the lung and liver tissues of mice experimentally infected with the influenza A/WSN/1/33(H1N1) virus indicated the presence of a large amount of the virus at all stages of the experiment that requires further research.

Yeast Surface-Displayed Quenchbody as a Novel Whole-Cell Biosensor for One-Step Detection of Influenza A (H1N1) Virus.

Timely surveillance of airborne pathogens is essential to preventing the spread of infectious diseases and safeguard human health. Methods for sensitive, efficient, and cost-effective detection of airborne viruses are needed. With advances in synthetic biology, whole-cell biosensors have emerged as promising platforms for environmental monitoring and medical diagnostics. However, the current design paradigm of whole-cell biosensors is mostly based on intracellular detection of analytes that can transport across the cell membrane, which presents a critical challenge for viral pathogens and large biomolecules. To address this challenge, we developed a new type of whole-cell biosensor by expressing and displaying VHH-based quenchbody (Q-body) on the surface of the yeast Saccharomyces cerevisiae for simple one-step detection of influenza A (H1N1) virus. Seventeen VHH antibody fragments targeting the hemagglutinin protein H1N1-HA were displayed on the yeast cells and screened for the H1N1-HA binding affinity. The functionally displayed VHHs were selected to create surface-displayed Q-body biosensors. The surface-displayed Q-body exhibiting the highest quenching and dequenching efficiency was identified. The biosensor quantitatively detected H1N1-HA in a range from 0.5 to 16 μg/mL, with a half-maximal concentration of 2.60 μg/mL. The biosensor exhibited high specificity for H1N1-HA over other hemagglutinin proteins from various influenza A virus subtypes. Moreover, the biosensor succeeded in detecting the H1N1 virus at concentrations from 2.4 × 104 to 1.5 × 107 PFU/mL. The results from this study demonstrated a new whole-cell biosensor design that circumvents the need for transport of analytes into biosensor cells, enabling efficient detection of the target virus particles.

Highly pathogenic avian influenza A virus subtype H5N1 (clade 2.3.4.4b) isolated from a natural protected area in Peru.

The panzootic caused by H5N1 avian influenza viruses is a high concern for wild birds' conservation and the study of spillover events into mammals. The near coding-complete genome of H5N1 clade 2.3.3.4b sequencing in the Miseq Illumina platform was performed from a bird located in Pantanos of Villa National Wildlife Refuge.