Doomed drones? Using passage experiments and mathematical modelling to determine Deformed wing virus population dynamics in male honey bees

Funding: This research was funded by the BBSRC (Biotechnology and Biological Sciences Research Council), grant numbers: BB/M010996/1 and BB/R00305X/1. P.C.S was supported by the BBSRC, grant number: BB/S00243X/1.Varroa destructor is an ectoparasitic mite of honeybees which vectors a range of pathogenic viruses, the most notable being Deformed wing virus (DWV). Mites parasitise bees during pupal development and male honeybees, drones, have a longer development cycle than female workers (24 versus 21 days), allow for more progeny mites to develop per foundress (1.6–2.5 compared to 0.7–1.45). How this longer exposure time influences evolution of the transmitted virus population is unknown. Using uniquely tagged viruses recovered from cDNA we investigated the replication, competition and morbidity of DWV genotypes in drones. Assays examining virus replication and morbidity revealed drones are highly susceptible to both predominant genotypes of DWV. In virus passage studies using an equimolar inocula of major DNA genotypes and their recombinants, the recombinant form dominated but did not reach 100% of the virus population within 10 passages. Using an in-silico model of the virus–mite–bee system we examined bottlenecks during virus acquisition by the mite and subsequent injection of viruses into the host, which may play a significant role in shaping virus diversity. This study furthers our understanding of the variables influencing DWV diversity changes and provides insight into areas of future research in the mite–virus–bee system.Publisher PDFPeer reviewe

All together now : geographically coordinated miticide treatment benefits honey bee health

This research was funded by the BBSRC (Biotechnology and Biological Sciences Research Council), grant number: BB/M010996/1.Deformed wing virus (DWV) is a pathogenic virus of honey bees transmitted by the ectoparasitic mite Varroa destructor. Annual overwintering colony losses, accounting for ~25% of all colonies, are associated with high levels of Varroa-DWV infestation. Effective miticide treatments are available to control Varroa. However, the absence of coordinated treatment means environmental transmission of mites continues unchecked. We aimed to determine whether rational, coordinated treatment is beneficial, and characterized the DWV population as an indicator of colony health. This study uses coordinated treatment of Varroa in a geographically isolated environment (Isle of Arran, Scotland) over 3 years. The study area contained 50–84 colonies managed by ~20 amateur beekeepers. Sampling and virus analysis to assess strain diversity and viral loads were conducted before and after treatments, and changes in population diversity were quantified by sequence analysis. Over the 3 years analysis of the virus population revealed that the dominant DWV variant shifted from Type A to Type B in all apiaries, regardless of mite levels or proximity to other colonies. During this period the number of managed colonies increased by 47% (57–84 colonies), but despite this, we estimate total mite numbers decreased by 58%. Synthesis and applications. In this study, the beekeepers in Arran significantly improved the number of colonies they managed, without importing any bees onto the island, indicating that an improved focus on management techniques, through the combination of a coordinated miticide programme and an improved understanding of bee diseases, could yield positive results for bee health and sustainability.Publisher PDFPeer reviewe

First come, first served : superinfection exclusion in Deformed wing virus is dependent upon sequence identity and not the order of virus acquisition

Funding: Biotechnology and Biological Sciences Research Council (BBSRC), BB/M00337X/2.Deformed wing virus (DWV) is the most important globally distributed pathogen of honey bees and, when vectored by the ectoparasite Varroa destructor, is associated with high levels of colony losses. Divergent DWV types may differ in their pathogenicity and are reported to exhibit superinfection exclusion upon sequential infections, an inevitability in a Varroa-infested colony. We used a reverse genetic approach to investigate competition and interactions between genetically distinct or related virus strains, analysing viral load over time, tissue distribution with reporter gene-expressing viruses and recombination between virus variants. Transient competition occurred irrespective of the order of virus acquisition, indicating no directionality or dominance. Over longer periods, the ability to compete with a pre-existing infection correlated with the genetic divergence of the inoculae. Genetic recombination was observed throughout the DWV genome with recombinants accounting for ~2% of the population as determined by deep sequencing. We propose that superinfection exclusion, if it occurs at all, is a consequence of a cross-reactive RNAi response to the viruses involved, explaining the lack of dominance of one virus type over another. A better understanding of the consequences of dual- and superinfection will inform development of cross-protective honey bee vaccines and landscape-scale DWV transmission and evolution.Publisher PDFPeer reviewe

Imprecise recombinant viruses evolve via a fitness-driven, iterative process of polymerase template-switching events

Funding: This work was supported by the Biotechnology and Biological Sciences Research Council - https://bbsrc.ukri.org/ - (BB/M009343/1 to D.J.E) and an ISSF award from The Wellcome Trust - https://wellcome.org/ - to the BSRC, University of St Andrews. F.G.A was supported by a PhD studentship from the Ministry of Education, Government of Saudi Arabia. S.J was supported by a Microbiology Society - https://microbiologysociety.org/ - Harry Smith Vacation Studentship awarded to K.B.Viruses with positive-sense RNA genomes, such as poliovirus, have several mechanisms by which they evolve. One of these is the process of recombination involving the large-scale exchange of genetic information. Recombination occurs during replication when the viral polymerase, bound to the nascent RNA chain, switches from copying one genome to another. However, the polymerase does not always accurately switch between the two, resulting in sequence duplications or deletions, and genomes that are referred to as imprecise. Over multiple rounds of replication sequence duplications are lost and genomes are resolved to wild type length, but it is unclear how this occurs. Here we used synthetic polioviruses containing defined sequence duplications to determine that the genome population undergoes repeated rounds of recombination until sequence duplications are lost and viruses with precise, wild type length genomes are selected for. This selection is based on the overall fitness of the virus population, with less fit imprecise viruses evolving more quickly. Our study suggests that recombination is a continual process where virus fitness drives the selection of a small subset of recombinant variants. These data are important for understanding how novel viruses evolve via recombination and how this process can be blocked to prevent novel and dangerous pathogens from arising.Publisher PDFPeer reviewe

Can plastic pollution drive the emergence and dissemination of novel zoonotic diseases?

As the volume of plastic in the environment increases, so too does human interactions with plastic pollution. Similarly, domestic, feral, and wild animals are increasingly interacting with plastic pollution, highlighting the potential for contamination of plastic wastes with animal faeces, urine, saliva, and blood. Substantial evidence indicates that once in the environment, plastics rapidly become colonised by microbial biofilm (the so-called ‘plastisphere), which often includes potentially harmful microbial pathogens (including pathogens that are zoonotic in nature). Climate change, increased urbanisation, and the intensification of agriculture, mean that the three-way interactions between humans, animals, and plastic pollution are becoming more frequent, which is significant as almost 60% of emerging human infectious diseases during the last century have been zoonotic. Here, we critically review the potential for contaminated environmental plastics to facilitate the evolution of novel pathogenic strains of microorganisms, and the subsequent role of plastic pollution in the cyclical dissemination of zoonotic pathogens. As the interactions between humans, animals, and plastic pollution continues to grow, and the volume of plastics entering the environment increases, there is clearly an urgent need to better understand the role of plastic waste in facilitating zoonotic pathogen evolution and dissemination, and the effect this can have on environmental and human health

Elevated immune gene expression is associated with poor reproductive success of urban blue tits

Urban and forest habitats differ in many aspects that can lead to modifications of the immune system of wild animals. Altered parasite communities, pollution, and artificial light at night in cities have been associated with exacerbated inflammatory responses, with possibly negative fitness consequences, but few data are available from free-living animals. Here, we investigate how urbanization affects major immune pathways and experimentally test potentially contributing factors in blue tits (Cyanistes caeruleus) from an urban and forest site. We first compared breeding adults by quantifying the mRNA transcript levels of proteins associated with anti-bacterial, anti-malarial (TLR4, LY86) and anti-helminthic (Type 2 transcription factor GATA3) immune responses. Adult urban and forest blue tits differed in gene expression, with significantly increased TLR4 and GATA3, but not LY86, in the city. We then experimentally tested whether these differences were environmentally induced by cross-fostering eggs between the sites and measuring mRNA transcripts in nestlings. The populations differed in reduced reproductive success, with a lower fledging success and lower fledgling weight recorded at the urban site. This mirrors the findings of our twin study reporting that the urban site was severely resource limited when compared to the forest. Because of low urban survival, robust gene expression data were only obtained from nestlings reared in the forest. Transcript levels in these nestlings showed no (TLR4, LY86), or weak (GATA3), differences according to their origin from forest or city nests, suggesting little genetic or maternal contribution to nestling immune transcript levels. Lastly, to investigate differences in parasite pressure between urban and forest sites, we measured the prevalence of malaria in adult and nestling blood. Prevalence was invariably high across environments and not associated with the transcript levels of the studied immune genes. Our results support the hypothesis that inflammatory pathways are activated in an urban environment and suggest that these differences are most likely induced by environmental factors

Vector species-specific association between natural Wolbachia infections and avian malaria in black fly populations

Thanks to the Institute of Biodiversity Animal Health and Comparative Medicine (University of Glasgow) for funding vector traps.Artificial infection of mosquitoes with the endosymbiont bacteria Wolbachia can interfere with malaria parasite development. Therefore, the release of Wolbachia-infected mosquitoes has been proposed as a malaria control strategy. However, Wolbachia effects on vector competence are only partly understood, as indicated by inconsistent effects on malaria infection reported under laboratory conditions. Studies of naturally-occurring Wolbachia infections in wild vector populations could be useful to identify the ecological and evolutionary conditions under which these endosymbionts can block malaria transmission. Here we demonstrate the occurrence of natural Wolbachia infections in three species of black fly (genus Simulium), which is a main vector of the avian malaria parasite Leucocytozoon. Prevalence of Leucocytozoon was high (25%), but the nature and magnitude of its association with Wolbachia differed between black fly species. Wolbachia infection was positively associated with avian malaria infection in S. cryophilum, negatively associated in S. aureum, and unrelated in S. vernum. These differences suggest that Wolbachia interacts with the parasite in a vector host species-specific manner. This provides a useful model system for further study of how Wolbachia influences vector competence. Such knowledge, including the possibility of undesirable positive association, is required to guide endosymbiont based control methods.Publisher PDFPeer reviewe

Elevated immune gene expression is associated with poor reproductive success of urban blue tits

Urban and forest habitats differ in many aspects that can lead to modifications of the immune system of wild animals. Altered parasite communities, pollution, and artificial light at night in cities have been associated with exacerbated inflammatory responses, with possibly negative fitness consequences, but few data are available from free-living animals. Here, we investigate how urbanization affects major immune pathways and experimentally test potentially contributing factors in blue tits (Cyanistes caeruleus) from an urban and forest site. We first compared breeding adults by quantifying the mRNA transcript levels of proteins associated with anti-bacterial, anti-malarial (TLR4, LY86) and anti-helminthic (Type 2 transcription factor GATA3) immune responses. Adult urban and forest blue tits differed in gene expression, with significantly increased TLR4 and GATA3, but not LY86, in the city. We then experimentally tested whether these differences were environmentally induced by cross-fostering eggs between the sites and measuring mRNA transcripts in nestlings. The populations differed in reduced reproductive success, with a lower fledging success and lower fledgling weight recorded at the urban site. This mirrors the findings of our twin study reporting that the urban site was severely resource limited when compared to the forest. Because of low urban survival, robust gene expression data were only obtained from nestlings reared in the forest. Transcript levels in these nestlings showed no (TLR4, LY86), or weak (GATA3), differences according to their origin from forest or city nests, suggesting little genetic or maternal contribution to nestling immune transcript levels. Lastly, to investigate differences in parasite pressure between urban and forest sites, we measured the prevalence of malaria in adult and nestling blood. Prevalence was invariably high across environments and not associated with the transcript levels of the studied immune genes. Our results support the hypothesis that inflammatory pathways are activated in an urban environment and suggest that these differences are most likely induced by environmental factors.Publisher PDFPeer reviewe

Hybridisation rates, population structure, and dispersal of sambar deer (Cervus unicolor) and rusa deer (Cervus timorensis) in south-eastern Australia

Context. Introduced populations of sambar deer (Cervus unicolor) and rusa deer (Cervus timorensis) are present across south-eastern Australia and are subject to local population control to alleviate their negative impacts. For management to be effective, identification of dispersal capability and management units is necessary. These species also readily hybridise, so additional investigation of hybridisation rates across their distributions is necessary to understand the interactions between the two species. Aims. Measure the hybridisation rate of sambar and rusa deer, assess broad-scale population structure present within both species and identify distinct management units for future population control, and measure the likely dispersal capability of both species. Methods. In total, 198 sambar deer, 189 rusa deer, and three suspected hybrid samples were collected across Victoria and New South Wales (NSW). After sequencing and filtering, 14 099 polymorphic single-nucleotide polymorphism (SNP) markers were retained for analysis. Hybridisation rates were assessed before the data were split by species to identify population structure, diversity indices, and dispersal distances. Key results. Across the entire dataset, 17 hybrids were detected. Broad-scale population structure was evident in sambar deer, but not among the sites where rusa deer were sampled. Analysis of dispersal ability showed that a majority of deer movement occurred within 20 km in both species, suggesting limited dispersal. Conclusions. Distinct management units of sambar deer can be identified from the dataset, allowing independent population control. Although broad-scale population structure was not evident in the rusa deer populations, dispersal limits identified suggest that rusa deer sites sampled in this study could be managed separately. Sambar × rusa deer hybrids are present in both Victoria and NSW and can be difficult to detect on the basis of morphology alone. Implications. Genetic analysis can identify broad-scale management units necessary for population control, and estimates of dispersal capability can assist in delineating management units where broad-scale population structure may not be apparent. The negative impacts associated with hybridisation require further investigation to determine whether removal of hybrids should be considered a priority management aim. © 2023 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Toxigenic Vibrio cholerae can cycle between environmental plastic waste and floodwater: Implications for environmental management of cholera

Globally, there has been a significant rise in cholera cases and deaths, with an increase in the number of low- and middle-income countries (LMICs) reporting outbreaks. In parallel, plastic pollution in LMICs is increasing, and has become a major constituent of urban dump sites. The surfaces of environmental plastic pollution can provide a habitat for complex microbial biofilm communities; this so-called 'plastisphere' can also include human pathogens. Under conditions simulating a peri-urban environmental waste pile, we determine whether toxigenic Vibrio cholerae (O1 classical; O1 El Tor; O139) can colonise and persist on plastic following a simulated flooding event. Toxigenic V. cholerae colonized and persisted on plastic and organic waste for at least 14 days before subsequent transfer to either fresh or brackish floodwater, where they can further persist at concentrations sufficient to cause human infection. Taken together, this study suggests that plastics in the environment can act as significant reservoirs for V. cholerae, whilst subsequent transfer to floodwaters demonstrates the potential for the wider dissemination of cholera. Further understanding of how diseases interact with plastic waste will be central for combating infection, educating communities, and diminishing the public health risk of plastics in the environment