Four distinct species of human coronaviruses (HCoVs) circulate in humans. Despite the recent attention due to SARS-CoV-2, a comprehensive understanding of the molecular epidemiology and genomic evolution of HCoVs remains unclear. Here, we employed primary differentiated human nasal epithelial cells for the successful isolation and genome sequencing of HCoVs derived from two retrospective cohorts in Singapore and Tanzania. Phylodynamic inference shows that HCoV-229E and HCoV-OC43 were subject to stronger genetic drift and reduced purifying selection from the early 2000s onwards, primarily targeting spike Domain A and B. This resulted in increased lineage diversification, coinciding with a higher effective reproductive number (Re>1.0). However, HCoV-NL63 and HCoV-HKU1 experienced weaker genetic drift and selective pressure with prolonged regional persistence. Our findings suggest that HCoV-229E and HCoV-OC43 viruses are adept at generating new variants and achieving widespread intercontinental dissemination driven by continuous genetic drift, recombination, and complex migration patterns.
Abstract The neuraminidase (NA) and hemagglutinin (HA) are essential surface glycoproteins of influenza A virus (IAV). In this study, the evolution of subtype N2 NA paired with H1 and H3 subtype HA in swine was evaluated to understand if the genetic diversity of HA and NA were linked. Using time-scaled Bayesian phylodynamic analyses, the relationships of paired swine N2 with H1 or H3 from 2009 to 2018 were evaluated. These data demonstrated increased relative genetic diversity within the major N2 clades circulating in swine in the USA (N2.1998 between 2014 and 2017 and N2.2002 between 2010 and 2016). Preferential pairing was observed among specific NA and HA genetic clades. Gene reassortment between cocirculating influenza A strains resulted in novel pairings that persisted. The changes in genetic diversity in the NA gene were quantified using Bayesian phylodynamic analyses, and increases in diversity were observed subsequent to novel NA–HA reassortment events. The rate of evolution among NA–N2 clades and HA–H1 and HA–H3 clades were similar. Bayesian phylodynamic analyses demonstrated strong spatial patterns in N2 genetic diversity, but frequent interstate movement of rare N2 clades provided opportunity for reassortment and emergence of new N2–HA pairings. The frequent regional movement of pigs and their influenza viruses is an explanation for the documented patterns of reassortment and subsequent changes in gene diversity. The reassortment and evolution of NA and linked HA evolution may result in antigenic drift of both major surface glycoproteins, reducing vaccine efficacy, with subsequent impact on animal health.
This study evaluated influenza A virus (IAV) detection and genetic diversity over time, specifically at the human–swine interface in breeding and nursery farms. Active surveillance was performed monthly in five swine farms in the Midwest United States targeting the employees, the prewean piglets at sow farms, and the same cohort of piglets in downstream nurseries. In addition, information was collected at enrollment for each employee and farm to assess production management practices, IAV vaccination status, diagnostic procedures, and biosecurity. Farm employee and swine samples were screened by IAV reverse transcription real-time polymerase chain reaction (RT-rtPCR), followed by IAV subtyping RT-rtPCR and whole genome sequencing on PCR-positive samples. This study showed higher positivity of IAV RNA detection in nursery pigs compared to prewean pigs, and more whole genome sequences were also obtained in the nursery phase. Surveillance of farm employees revealed two detections of H3N2 representing the 2022–2023 human IAV season, confirming the presence of influenza in farm employees while present at work, and thus highlighting the importance of biosecurity measures at the human–swine interface. This study highlights the importance of routine active surveillance to understand the dynamics of IAV at the farm level in both farm employees and swine.
Porcine reproductive and respiratory syndrome virus (PRRSV) is the most economically important swine pathogen in North America and is second globally only to African swine fever virus. PRRSV is a positive sense, single stranded RNA virus associated with reproductive disorder of sows and respiratory disease of pigs at all age. Diagnostic tests are commonly used to monitor the presence of PRRSV in swine populations including sequencing the open reading frame 5 (ORF5) gene to track the epidemiology of the virus and lateral introductions into a farm. PRRSView is a web portal created at the Iowa State University Veterinary Diagnostic Laboratory (ISU VDL) to host analytical and phylogenetic tools related to PRRSV ORF5 sequences with the goal of assisting veterinarians and producers in evaluating the genetic diversity, spatial, and temporal aspects of ORF5 sequences maintained in the ISU VDL database. PRRSView works in conjunction with the broader Swine Disease Reporting System (SDRS) project to contextualize the ever changing patterns of PRRSV diversity, and supports interactive tools for veterinarians to analyze their sequence data compared to other sequences detected throughout the United States. The PRRSView homepage provides a phylogenetic overview of the sequences generated by the ISU VDL within the previous month, indicating the current strains detected in circulation. There are currently three ORF5 analytical tools available on PRRSView: a genetic sequence BLAST tool, a vaccine identity tool, and an RFLP tool. The ORF5 BLAST tool allows the users to submit their ORF5 gene sequences and returns up to 10 closely related sequences from the ISU VDL database, with metadata that includes the state, genetic lineage, RFLP, and identity to the query sequence. The vaccine identity tool allows users to quickly calculate the percent homology of their sequence(s) to five different PRRSV vaccines: Inglevac PRRS ATP, Inglevac PRRS MLV, Prime Pac PRRS, Fostera PRRS, and Prevacent PRRS, as well as the distance to the Lelystad strain. This tool also builds a neighbor joining tree with a set of curated strains to estimate the genetic lineage of the sequence, which is rendered in the web browser for viewing. Additionally, this tool will calculate the RFLP of the sequence, and the exact positions of the cut sites are shown when hovering over the RFLP value. The last analytical tool provided is the ORF5 RFLP tool, which quickly calculates the RFLP pattern of the input sequences. These analytical tools are designed to allow veterinarians and researchers to easily analyze their PRRSV ORF5 sequences against the expansive ISU VDL database to gain valuable epidemiologic information and comparative data regarding the genetic lineages and related metadata of the PRRSV circulating in a production system, while lowering the barrier of entry for use.
Effective biosecurity practices in swine production are key in preventing the introduction and dissemination of infectious pathogens. Ideally, on-farm biosecurity practices should be chosen by their impact on bio-containment and bio-exclusion; however, quantitative supporting evidence is often unavailable. Therefore, the development of methodologies capable of quantifying and ranking biosecurity practices according to their efficacy in reducing disease risk has the potential to facilitate better-informed choices of biosecurity practices. Using survey data on biosecurity practices, farm demographics, and previous outbreaks from 139 herds, a set of machine learning algorithms were trained to classify farms by porcine reproductive and respiratory syndrome virus status, depending on their biosecurity practices and farm demographics, to produce a predicted outbreak risk. A novel interpretable machine learning toolkit, MrIML-biosecurity, was developed to benchmark farms and production systems by predicted risk and quantify the impact of biosecurity practices on disease risk at individual farms. By quantifying the variable impact on predicted risk, 50% of 42 variables were associated with fomite spread while 31% were associated with local transmission. Results from machine learning interpretations identified similar results, finding substantial contribution to predicted outbreak risk from biosecurity practices relating to the turnover and number of employees, the surrounding density of swine premises and pigs, the sharing of haul trailers, distance from the public road and farm production type. In addition, the development of individualized biosecurity assessments provides the opportunity to better guide biosecurity implementation on a case-by-case basis. Finally, the flexibility of the MrIML-biosecurity toolkit gives it the potential to be applied to wider areas of biosecurity benchmarking, to address biosecurity weaknesses in other livestock systems and industry-relevant diseases.
During the SARS-CoV-2 pandemic, we have witnessed the emergence and disappearance of variants. When random mutations generate a new variant capable of infecting more individuals than existing variants, such a variant poses a public health threat due to the virus’s increased transmissibility. Therefore, it is essential to know how transmissible a new variant is compared to existing ones. In this paper, we introduce a computer program called RelRe, which allows users to estimate the relative instantaneous reproduction numbers among variants as well as the relative generation time using time series data of variant counts. Based on the estimated parameters, one can predict future variant replacements. The program was implemented with the Julia language, and its source code is available on our GitHub page (https://github.com/KimihitoIto/RelRe).