Biology subjects

Raghwani, J.

Publications and source records attributed to Raghwani, J..

13 recordsLinked to original sources

bioRxiv2025

Prevalence of zoonotic hepatic nematode varies with small mammal community diversity across a heterogenous landscape in Eastern Uganda

Identifying key drivers of pathogen infection prevalence and intensity in wildlife is essential to understand disease dispersal and transmission. Calodium hepatica (syn. Capillaria hepatica) is a generalist nematode that infects liver parenchyma of mammals worldwide and is capable of human infections. Prevalence ranges from 0-100% in wildlife, often varying across small geographic areas, making it an ideal parasite for understanding ecological drivers of variation. Here, we quantify prevalence of Calodium hepatica and present initial surveys of synanthropic small mammals in four villages representing differing land cover. Cross-sectional rodent trapping was conducted within and around households over consecutive dry seasons in Eastern Uganda. 18s rRNA gene of C. hepatica was amplified and a sub-set of PCR products sequenced to confirm presence of C. hepatica. Landscape structural diversity was classified by tree crown density and mean canopy height derived from 30m LiDAR data within a 0.5km buffer. Multivariable binomial generalised linear models were fit to C. hepatica prevalence. C. hepatica infection was common (overall 34.5%, CI95% 27.9-41.0) and found in rodent and shrew species inside and outside residences. We observe village-level differences in prevalence (18.2%-75.0%), with higher C. hepatica prevalence associated with higher relative proportion of Rattus rattus to other species (aOR=2.22 CI95% 1.30-3.85). Host diversity appears to be protective against parasite prevalence. Differences in molecular and macroscopy identification highlight challenges in diagnosis and a need for more specialized molecular tools. Further investigation is required to understand individual host and community variation in pathogen infection intensity and implications for zoonotic risk. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=127 SRC="FIGDIR/small/674808v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@e9cfdaorg.highwire.dtl.DTLVardef@4d0f6dorg.highwire.dtl.DTLVardef@12fe43dorg.highwire.dtl.DTLVardef@1318d83_HPS_FORMAT_FIGEXP M_FIG C_FIG Key FindingsO_LIZoonotic reservoirs, including Rattus rattus, Mastomys erythroleucus and Crocidura olivieri, are abundant in human-modified habitats and show elevated levels of household incursion C_LIO_LIHigh prevalence of Calodium hepatica, a nematode endoparasite, was identified in a range of small mammal species C_LIO_LIHigher proportion of native small mammal species relative to Rattus rattus appears protective against zoonotic pathogen load, with higher village-level C. hepatica prevalence in ecologically depleted sites C_LIO_LISmall-holder agriculture may provide a dilution effect through secondary wildlife support and small mammal competition, while dense village settings potentiate C. hepatica prevalence C_LI

ecology↗

bioRxiv2025

Quantifying virus load and characterising virus diversity in wildlife samples with target enrichment sequencing

Metagenomics is a powerful tool for characterising viruses, with broad applications across diverse disciplines, from understanding the ecology and evolutionary history of viruses to identifying causative agents of emerging outbreaks with unknown aetiology. Additionally, metagenomic data contains valuable information about the amount of virus present within samples. However, we have yet to leverage metagenomics to assess viral load, which is a key epidemiological parameter. To effectively use sequencing outputs to inform transmission, we need to understand the relationship between read depth and viral load across a diverse set of viruses. Here, using target enrichment sequencing, we investigated the detection and recovery of virus genomes by spiking known concentrations of DNA and RNA viruses into wild rodent faecal samples. In total, 15 experimental replicates were sequenced with target enrichment sequencing and compared to shotgun sequencing of the same background samples. Target enriched sequencing recovered all spike-in viruses at every concentration (102, 103, and 105 {+/-} 1 log genome copies) and showed a log-linear relationship between spike-in concentration and mean read depth. Background viruses (including Kobuvirus and Cardiovirus) were recovered consistently across all biological and technical replicates, but genome coverage was variable between virus genera and likely reflected the composition of target enrichment probe panel. Overall, our study highlights the strengths and weaknesses of using commercially available panels to quantify and characterise wildlife viromes, and underscores the importance of probe panel design for accurately interpreting coverage and read depth. To advance the use of metagenomics for understanding virus transmission, further research will be needed to elucidate how sequencing strategy (e.g. library depth, pooling), virome composition, and probe design influence viral read counts and genome coverage.

genomics↗

bioRxiv2024

Utilising citizen science data to rapidly assess potential wild bridging hosts and reservoirs of infection: avian influenza outbreaks in Great Britain

High pathogenicity avian influenza virus (HPAIV) is a rapidly evolving orthomyxovirus causing significant economic and environmental harm. Wild birds are a key reservoir of infection and an important source of viral incursions into poultry populations. However, we lack thorough understanding of which wild species drive incursions and whether this changes over time. We explored associations between abundances of 152 avian species and cases of HPAI in poultry premises across Great Britain between October-2021 and January-2023. Spatial generalised additive models were used, with species abundance distributions sourced from eBird modelled predictions. Associations were investigated at the species-specific level and across aggregations of species. During autumn/winter, associations were generally strongest with waterbirds such as ducks and geese; however, we also found significant associations in other groups such as non-native gamebirds, and rapid change in species-specific associations over time. Our results demonstrate the value of citizen science in rapid exploration of wild reservoirs of infection as facilitators of disease incursion into domestic hosts, especially in regions where surveillance programmes in wild birds are absent. This can be a critical step towards improving species-specific biosecurity measures and targeted surveillance; particularly for HPAIV, which has undergone sudden shifts in host-range and continues to rapidly evolve.

ecology↗

bioRxiv2023

Association of poultry vaccination with the interspecies transmission and molecular evolution of H5 subtype avian influenza virus

The effectiveness of vaccinating poultry in preventing the transmission of highly pathogenic avian influenza viruses (AIVs) has been questioned for years and its impact on wild birds is uncertain 1-3. Here we reconstruct movements of H5 subtype AIV lineages among vaccinated poultry, unvaccinated poultry, and wild birds, worldwide from 1996 to 2023. We find that lineage transitions among host types are lagged and that movements from wild birds to unvaccinated poultry were more frequent than those from wild birds to vaccinated poultry. However, we also find that the HA gene of the AIV lineage that circulated predominately among Chinese poultry with high vaccination coverage underwent faster evolution and greater nonsynonymous divergence than other lineages. Further, this Chinese poultry lineage contained more codons inferred to be under positive selection, including at known antigenic sites, and its rates of nonsynonymous divergence and adaptative fixation increased after mass poultry vaccination began. Our results indicate that the epidemiological, ecological and evolutionary consequences of widespread AIV vaccination in poultry may be linked in complex ways, and that much work is needed to better understand how such interventions may affect AIV transmission to, within and from wild birds.

evolutionary biology↗

bioRxiv2022

Discovery of a novel coronavirus in Swedish bank voles (Myodes glareolus)

We identified a novel Betacoronavirus from bank voles (Myodes glareolus) in Grimso, Sweden. Repeated detection over three years and an overall prevalence of 3.4% suggests the virus commonly occurs in bank voles. Furthermore, phylogenetic analyses indicate the virus belongs to a highly divergent Embecovirus lineage predominantly associated with bank voles.

microbiology↗

bioRxiv2022

Seasonal dynamics of the wild rodent faecal virome

Viral discovery studies in wild animals often rely on cross-sectional surveys at a single time point. As a result, our understanding of the temporal stability of wild animal viromes remains poorly resolved. While studies of single host-virus systems indicate that host and environmental factors influence seasonal virus transmission dynamics, comparable insights for whole viral communities in multiple hosts are lacking. Leveraging non-invasive faecal samples from a long-term wild rodent study, we characterised viral communities of three common European rodent species (Apodemus sylvaticus, A. flavicollis, and M. glareolus) living in temperate woodland over a single year. Our findings indicate that a substantial fraction of the rodent virome is seasonally transient and associated with vertebrate or bacteria hosts. Further analyses of one of the most abundant virus families, Picornaviruses, show pronounced temporal changes in viral richness and diversity, which were associated with concurrent and up to [~]3-month lags in host density, ambient temperature, rainfall and humidity, suggesting complex feedbacks from the host and environmental factors on virus transmission and shedding in seasonal habitats. Overall, this study emphasizes the importance of understanding the seasonal dynamics of wild animal viromes in order to better predict and mitigate zoonotic risks.

ecology↗

bioRxiv2020

Limited evolution despite years of measurable viremia in a cART-treated seronegative HIV-1 positive individual

Understanding the role that antibodies play in controlling HIV-1 infection and in the dynamics that underpin the formation of the HIV-1 reservoir are important steps towards combatting this global disease. To address these gaps, we performed whole-genome, deep sequence analysis of longitudinal plasma HIV-1 samples from an individual who failed to develop detectable anti-HIV-1 antibodies for 4 years post infection. These analyses reveal limited evolution despite months of measurable viremia during treatment with cART. We used a mathematical model to simultaneously analyse the viral and evolutionary dynamics of this unique individual. We propose a role for antibodies in reducing viral infectivity and demonstrate how our data are consistent with a theory of rapid activation of latently infected cells prior to effective viral suppression. Our study supports and elucidates a recent finding that although the latent reservoir persists for years once virus is effectively suppressed, prior to suppression, viral strains within the reservoir turn over rapidly. The implications for a cure are significant.

evolutionary biology↗

bioRxiv2020

Vulnerabilities in coronavirus glycan shields despite extensive glycosylation

Severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronaviruses (CoVs) are zoonotic pathogens with high fatality rates and pandemic potential. Vaccine development has focussed on the principal target of the neutralizing humoral immune response, the spike (S) glycoprotein, which mediates receptor recognition and membrane fusion. Coronavirus S proteins are extensively glycosylated viral fusion proteins, encoding around 69-87 N-linked glycosylation sites per trimeric spike. Using a multifaceted structural approach, we reveal a specific area of high glycan density on MERS S that results in the formation of under-processed oligomannose-type glycan clusters, which was absent on SARS and HKU1 CoVs. We provide a comparison of the global glycan density of coronavirus spikes with other viral proteins including HIV-1 envelope, Lassa virus glycoprotein complex, and influenza hemagglutinin, where glycosylation plays a known role in shielding immunogenic epitopes. Consistent with the ability of the antibody-mediated immune response to effectively target and neutralize coronaviruses, we demonstrate that the glycans of coronavirus spikes are not able to form an efficacious high-density global shield to thwart the humoral immune response. Overall, our data reveal how differential organisation of viral glycosylation across class I viral fusion proteins influence not only individual glycan compositions but also the immunological pressure across the viral protein surface.

microbiology↗

bioRxiv2019

A de novo approach to inferring within-host fitness effects during untreated HIV-1 infection

In the absence of effective antiviral therapy, HIV-1 evolves in response to the within-host environment, of which the immune system is an important aspect. During the earliest stages of infection, this process of evolution is very rapid, driven by a small number of CTL escape mechanisms. As the infection progresses, immune escape variants evolve under reduced magnitudes of selection, while competition between an increasing number of polymorphic alleles (i.e., clonal interference) makes it difficult to quantify the magnitude of selection acting upon specific variant alleles. To tackle this complex problem, we developed a novel multi-locus inference method to evaluate the role of selection during the chronic stage of within-host infection. We applied this method to targeted sequence data from the p24 and gp41 regions of HIV-1 collected from 34 patients with long-term untreated HIV-1 infection. We identify a broad distribution of beneficial fitness effects during infection, with a small number of variants evolving under strong selection and very many variants evolving under weaker selection. The uniquely large number of infections analysed granted a previously unparalleled statistical power to identify loci at which selection could be inferred to act with statistical confidence. Our model makes no prior assumptions about the nature of alleles under selection, such that any synonymous or non-synonymous variant may be inferred to evolve under selection. However, the majority of variants inferred with confidence to be under selection were non-synonymous in nature, and in nearly all cases were associated with either CTL escape in p24 or neutralising antibody escape in gp41. Sites inferred to be under selection in multiple hosts have high within-host and between-host diversity albeit not all sites with high between-host diversity were inferred to be under selection at the within-host level. Our identification of selection at sites associated with resistance to broadly neutralising antibodies (bNAbs) highlights the need to fully understand the role of selection in untreated individuals when designing bNAb based therapies.\n\nAuthor SummaryDuring the within-host evolution of HIV-1, the diversity of the viral population increases, with many beneficial variants competing against each other. This competition, known as clonal interference, makes the identification of variants under positive selection a challenging task. We here apply a novel method for the inference of selection to targeted within-host sequence data describing changes in the p24 and gp41 genes during HIV-1 infection in 34 patients. Our method adopts a parsimonious approach, assigning selection to the smallest number of variants necessary to explain the evolution of the system. The large size of our dataset allows for the confident identification of variants under selection, alleles at certain loci being repeatedly inferred as under selection within multiple individuals. While early CTL escape mutations have been identified to evolve under strong positive selection, we identify a distribution of beneficial fitness effects in which a large number of mutations are under weak selection. Variants that were confidently identified under selection were primarily found to be associated with either CTL escape in p24 or neutralising antibody escape in gp41, including sites associated with escape from broadly neutralising antibodies. We find that the most frequently selected loci have high diversity both within-host and at the between-host level.

evolutionary biology↗

bioRxiv2018

Link between the numbers of particles and variants founding new HIV-1 infections depends on the timing of transmission

Understanding which HIV-1 variants are most likely to be transmitted is important for vaccine design and predicting virus evolution. Since most infections are founded by single variants, it has been suggested that selection at transmission has a key role in governing which variants are transmitted. We show that the composition of the viral population within the donor at the time of transmission is also important. To support this argument, we developed a probabilistic model describing HIV-1 transmission in an untreated population, and parameterised the model using both within-host next generation sequencing data and population-level epidemiological data on heterosexual transmission. The most basic HIV-1 transmission models cannot explain simultaneously the low probability of transmission and the non-negligible proportion of infections founded by multiple variants. In our model, transmission can only occur when environmental conditions are appropriate (e.g. abrasions are present in the genital tract of the potential recipient), allowing these observations to be reconciled. As well as reproducing features of transmission in real populations, our model demonstrates that, contrary to expectation, there is not a simple link between the number of viral variants and the number of viral particles founding each new infection. These quantities depend on the timing of transmission, and infections can be founded with small numbers of variants yet large numbers of particles. Including selection, or a bias towards early transmission (e.g. due to treatment) acts to enhance this conclusion. In addition, we find that infections initiated by multiple variants are most likely to have derived from donors with intermediate set-point viral loads, and not from individuals with high set-point viral loads as might be expected. We therefore emphasise the importance of considering viral diversity in donors, and the timings of transmissions, when trying to discern the complex factors governing single or multiple variant transmission.

epidemiology↗

bioRxiv2018

High resolution evolutionary analysis of within-host hepatitis C virus infection

Despite the breakthroughs in the treatment of HCV infection in recent years, we have a limited understanding of how virus diversity generated within individuals impacts the evolution and spread of HCV variants at the population scale. Addressing this gap will be important for building models for molecular epidemiology, which can identify main sources of disease transmission and evaluate the risks of drug-resistance mutations emerging and disseminating in a population. Here, we have undertaken a high-resolution analysis of HCV within-host evolution from four individuals co-infected with HIV. Specifically, we used long-read, deep-sequenced data of the full-length HCV envelope glycoprotein, longitudinally sampled from acute to chronic HCV infection to investigate the underlying viral evolutionary dynamics. In three individuals we found strong statistical support for population structure maintaining within-host HCV genetic diversity. Furthermore, we found significant variation in rates of molecular evolution among different regions of the HCV envelope region, both within and between individuals. Lastly, we report the first estimate of the within-host population genetic rate of recombination for HCV (0.28 x 10-7 recombinations per site per day; interquartile range: 0.13-1.05 x 10-7), which is two orders of magnitude lower than that estimated for HIV-1, and four orders of magnitude lower than the nucleotide substitution rate of the HCV envelope gene. Together, these observations indicate that population structure and strong genetic linkage shapes within-host HCV evolutionary dynamics. These results will guide the future investigation of potential HCV drug resistance adaptation during infection, and at the population scale.

evolutionary biology↗

bioRxiv2017

Selection on non-antigenic gene segments of seasonal influenza A virus and its impact on adaptive evolution

Most studies on seasonal influenza A/H3N2 virus adaptation have focused on the main antigenic gene, haemagglutinin. However, there is increasing evidence that the genome-wide genetic background of novel antigenic variants can influence these variants emergence probabilities and impact their patterns of dominance in the population. This suggests that non-antigenic genes may be important in shaping the viral evolutionary dynamics. To better understand the role of selection on non-antigenic genes in the adaptive evolution of seasonal influenza viruses, we here develop a simple population genetic model that considers a virus with one antigenic and one non-antigenic gene segment. By simulating this model under different regimes of selection and reassortment, we find that the empirical patterns of lineage turnover for the antigenic and non-antigenic gene segments are best captured when there is both limited viral coinfection and selection operating on both gene segments. In contrast, under a scenario of only neutral evolution in the non-antigenic gene segment, we see persistence of multiple lineages for long periods of time in that segment, which is not compatible with the observed molecular evolutionary patterns. Further, we find that reassortment, occurring in coinfected individuals, can increase the speed of viral adaptive evolution by primarily reducing selective interference and genetic linkage effects mediated by the non-antigenic gene segment. Together, these findings suggest that, for influenza, with 6 internal or non-antigenic gene segments, the evolutionary dynamics of novel antigenic variants are likely to be influenced by the genome-wide genetic background as a result of linked selection among both beneficial and deleterious mutations.

evolutionary biology↗

bioRxiv2017

Molecular evolution, diversity and adaptation of H7N9 influenza A viruses in China

A novel H7N9 avian influenza virus has caused five human epidemics in China since 2013. The substantial increase in prevalence and the emergence of antigenically divergent or highly pathogenic (HP) H7N9 strains during the current outbreak raises concerns about the epizootic-potential of these viruses. Here, we investigate the evolution and adaptation of H7N9 by combining publicly available data with newly generated virus sequences isolated in Guangdong between 2015-2017. Phylogenetic analyses show that currently-circulating H7N9 viruses belong to distinct lineages with differing spatial distributions. Using ancestral sequence reconstruction and structural modelling we have identified parallel amino-acid changes on multiple separate lineages. Furthermore, we infer mutations in HA primarily occur at sites involved in receptor-recognition and/or antigenicity. We also identify seven new HP strains, which likely emerged from viruses circulating in eastern Guangdong around March 2016 and is further associated with a high rate of adaptive molecular evolution.

evolutionary biology↗