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bioRxiv2021

Natural infection of SARS-CoV-2 delta variant in Asiatic lions (Panthera leo persica) in India

In May 2021, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was detected in nine Asiatic lions (Panthera leo persica) in Arignar Anna Zoological Park, Chennai, Tamil Nadu, India. Sequence and phylogenetic analysis showed that the SARS-CoV-2 viruses belong to a variant of concern (VOC, delta variant, B.1.617.2 lineage) and that these viruses clustered with B.1.617.2 lineage viruses of the same geographical region detected in the same month.

microbiology↗

bioRxiv2021

Measuring biodiversity from DNA in the air

Impacts of the biodiversity crisis far exceed our ability to monitor changes in terrestrial ecosystems. Environmental DNA has revolutionized aquatic biomonitoring, permitting remote population and diversity assessments. Here we demonstrate that DNA from terrestrial animals can now be collected from the air under natural conditions, a ground-breaking advance for terrestrial biomonitoring. Using air samples from a zoological park, where species are spatially confined and unique compared to native fauna, we show that DNA in air can be used to identify the captive species and their potential interactions with local taxa. Air samples contained DNA from 25 species of mammal and bird including 17 known (and distinct) terrestrial zoo species. We also identified food items from air sampled in enclosures and detected four taxa native to the local area, including the Eurasian hedgehog, endangered in the UK, and the muntjac deer, a locally established invasive species. Our data provide evidence that airDNA is concentrated around recently inhabited areas (e.g., indoor enclosures) but that there is dispersal away from the source suggesting an ecology to airDNA movement which highlights the potential for airDNA sampling at distance. Our data clearly demonstrate the profound potential of air as a source of DNA for global terrestrial biomonitoring and ecological analysis. Significance StatementThe global decline in biodiversity requires rapid non-invasive biomonitoring tools applicable at a global scale. In this study we collect environmental DNA from mammals and birds from air samples collected in a natural setting. Using only air, we identified 25 species of mammal and bird known to be in the area. Our dataset detected species at risk of local extinction and several confirmed predator-prey interactions. This approach will revolutionize terrestrial biodiversity surveys.

ecology↗

bioRxiv2021

Revisiting rustrela virus - new cases of encephalitis and a solution to the capsid enigma

Rustrela virus (RusV, species Rubivirus strelense) is a recently discovered relative of rubella virus (RuV) that has been detected in cases of encephalitis across a wide spectrum of mammals, including placental and marsupial animals. Here we diagnosed two additional cases of fatal RusV-associated meningoencephalitis in a South American coati (Nasua nasua) and a Eurasian otter (Lutra lutra) that were detected in a zoological garden with history of prior RusV infections. Both animals showed abnormal movement or unusual behaviour and their brains tested positive for RusV using specific RT-qPCR and RNA in situ hybridization. As previous sequencing of RusV proved to be very challenging, we employed a sophisticated target-specific capture enrichment with specifically designed RNA baits to generate complete RusV genome sequences from both detected encephalitic animals and apparently healthy wild yellow-necked field mice (Apodemus flavicollis). Furthermore, the technique was used to revise three previously published RusV genomes from two encephalitic animals and a wild yellow-necked field mouse. Virus-to-host sequence ratio and thereby sequence coverage improved markedly using the enrichment method as compared to standard procedures. When comparing the newly generated RusV sequences to the previously published RusV genomes, we identified a previously undetected stretch of 309 nucleotides predicted to represent the intergenic region and the sequence encoding the N-terminus of the capsid protein. This indicated that the original RusV sequence was likely incomplete due to misassembly of the genome at a region with an exceptionally high G+C content of >80 mol%, which could not be resolved even by enormous sequencing efforts with standard methods. The updated capsid protein amino acid sequence now resembles those of RuV and ruhugu virus in size and harbours a predicted RNA binding domain that was not encoded in the original RusV genome version. The new sequence data indicate that RusV has the largest overall genome (9,631 nucleotides), intergenic region (290 nucleotides) and capsid protein-encoding sequence (331 codons) within the genus Rubivirus.

microbiology↗

bioRxiv2022

A developmental program that regulates mammalian organ size offsets evolutionary distance

Pigs are evolutionarily more distant from humans than mice, but their physiological organs are closest to humans. The molecular program leading to a more than 1,000-fold increase in organ size in pigs and humans over that of mice across evolution has not been elucidated. We generated large-scale transcriptional landscapes throughout swine lung development. Our cross-species single-cell molecular atlas let us discover swine progenitor identities, stage-specific markers, and a core organ-size regulation program (COSRP), well-conserved in swine and humans but less so in mice. Across eight mammalian species, human COSRP promoters showed higher homologies to evolutionary-distant large animals, including pigs, than evolutionary-close small animals. Our study provides a molecular foundation during swine lung development that unveils animal size regulation conserved in the COSRP promoter, independent of genome-wide evolution. COSRP is a critical paradigm for studying thousands-fold changes in biological sizes in evolution, development, cancer, zoology, respirology, organoids, and biotechnology, particularly human-compatible organ generation. One Sentence SummaryA cross-species developmental molecular atlas identified the indicator of lung and animal size beyond evolution

evolutionary biology↗

bioRxiv2023

All Quiet on the Western Front? The evolutionary history of monogeneans (Dactylogyridae: Cichlidogyrus, Onchobdella) infecting a West and Central African tribe of cichlid fishes (Chromidotilapiini)

Owing to the largely unexplored diversity of metazoan parasites, their speciation mechanisms and the circumstances under which such speciation occurs--in allopatry or sympatry--remain vastly understudied. Cichlids and their monogenean flatworm parasites have previously served as study system for macroevolutionary processes, e.g. for the role of East African host radiations on parasite communities. Here, we investigate the diversity and evolution of the poorly explored monogeneans infecting a West and Central African lineage of cichlid fishes: Chromidotilapiini Greenwood, 1987, which is the most species-rich tribe of cichlids in this region. We screened gills of 149 host specimens (27 species) from natural history collections and measured systematically informative characters of the sclerotised attachment and reproductive organs of the parasites. Ten monogenean species (Dactylogyridae: Cichlidogyrus and Onchobdella) were found, eight of which are described and one redescribed herein. The phylogenetic positions of chromidotilapiines-infecting species of Cichlidogyrus were inferred through a parsimony analysis of the morphological characters. Furthermore, we employed machine learning algorithms to detect morphological features associated with the main lineages of Cichlidogyrus. Although the results of these experimental algorithms remain inconclusive, the parsimony analysis indicates that West and Central African lineages of Cichlidogyrus and Onchobdella are monophyletic, unlike the paraphyletic host lineages. Several instances of host sharing suggest occurrences of intra-host speciation (sympatry) and host switching (allopatry). Some morphological variation was recorded that may also indicate the presence of species complexes. We conclude that collection material can provide important insights on parasite evolution despite the lack of well-preserved DNA material. Data availability statementType material was deposited in the invertebrate collection of the Royal Museum for Central Africa (Tervuren, Belgium) (RMCA) (RMCA_VERMES_XXXXX-XXX), the collection of the Research Group Zoology: Biodiversity and Toxicology of Hasselt University (Diepenbeek, Belgium) (HU XXX-XXX), the Finnish Museum of Natural History (Helsinki, Finland) (MZH XXXXX-XXX), and the Iziko South African Museum (Cape Town, South Africa) (SAMC-XXXXXXX-XXX). The morphological data that support the findings of this study are openly available in MorphoBank at www.morphobank.org, at https://dx.doi.org/XXXXXXXX. Phylogenetic trees and data matrices for the analysis in TNT are included as additional data in MorphoBank.

evolutionary biology↗

bioRxiv2023

Molecular evidence of anteroposterior patterning in adult echinoderms

The origin of the pentaradial body plan of echinoderms from a bilateral ancestor is one of the most enduring zoological puzzles. Since echinoderms are defined by morphological novelty, even the most basic axial comparisons with their bilaterian relatives are problematic. Here, we used conserved antero-posterior (AP) axial molecular markers to determine whether the highly derived adult body plan of echinoderms masks underlying patterning similarities with other deuterostomes. To revisit this classical question, we used RNA tomography and in situ hybridizations in the sea star Patiria miniata to investigate the expression of a suite of conserved transcription factors with well-established roles in the establishment of AP polarity in bilaterians. We find that the relative spatial expression of these markers in P. miniata ambulacral ectoderm shows similarity with other deuterostomes, with the midline of each ray representing the most anterior territory and the most lateral parts exhibiting a more posterior identity. Interestingly, there is no ectodermal territory in the sea star that expresses the characteristic bilaterian trunk genetic patterning program. This suggests that from the perspective of ectoderm patterning, echinoderms are mostly head-like animals, and prompts a reinterpretation of the evolutionary trends that made echinoderms the most derived animal group.

developmental biology↗

bioRxiv2023

Life history and cancer in birds: clutch size predicts cancer

Cancer is a disease that affects nearly all multicellular life, including birds. However, little is known about what factors explain the variance in cancer prevalence among species. Litter size is positively correlated with cancer prevalence in managed species of mammals, and larger body size, but not incubation or nestling period, is linked to tumor prevalence in wild birds. Also, birds that produce more elaborate sexual traits are expected to have fewer resources for cancer defenses and thus higher cancer prevalence. In this study, we examined whether cancer prevalence is associated with a wide variety of life history traits (clutch size, incubation length, body mass, lifespan, and the extent of sexual dimorphism) across 108 species of managed birds in 25 different zoological facilities, sanctuaries, and veterinary clinics. We found that clutch size was positively correlated with cancer and neoplasia (both benign and malignant) prevalence, even after controlling for body mass. Cancer prevalence was not associated with incubation length, body mass, lifespan, or sexual dimorphism. The positive correlations of clutch size with cancer prevalence and neoplasia prevalence suggest that there may be life-history trade-offs between reproductive investment and somatic maintenance (in the form of cancer prevention mechanisms) in managed birds.

cancer biology↗

bioRxiv2023

Rapid flapping and fiber-reinforced membrane wings are key to high-performance bat flight

Bats fly using significantly different wing motions than other fliers, stemming from the complex interplay of their membrane wings motion and structural properties. Biological studies show that many bats fly at Strouhal numbers, the ratio of flapping to flight speed, 50-150% above the range typically associated with optimal locomotion. We use high-resolution fluid-structure interaction simulations of a bat wing to independently study the role of kinematics and material/structural properties on aerodynamic performance and show that peak propulsive and lift efficiencies for a bat-like wing motion require flapping 66% faster than for a symmetric motion, agreeing with the increased flapping frequency observed in zoological studies. In addition, we find that reduced membrane stiffness is associated with improved propulsive efficiency until the membrane flutters, but that incorporating microstructural anisotropy arising from biological fiber reinforcement enables a tenfold reduction of the flutter energy whilst maintaining high aerodynamic efficiency. Our results indicate that animals with specialized flapping motions may have correspondingly specialized flapping speeds, in contrast to arguments for a universally efficient Strouhal range. Additionally, our study demonstrates the significant role that the microstructural constitutive properties of the membrane wing of a bat can have on its propulsive performance.

biophysics↗

bioRxiv2023

High pathogenicity avian influenza A (H5N1) clade 2.3.4.4b virus infection in a captive Tibetan black bear (Ursus thibetanus): investigations based on paraffin-embedded tissues, France, 2022

High pathogenicity avian influenza viruses H5Nx (HPAIVs) of clade 2.3.4.4b have been circulating increasingly in both wild and domestic birds in recent years. In turn, this has led to an increase in the number of spillovers events affecting mammals. In November 2022, a HPAIV H5N1 caused an outbreak in a zoological park in the south of France, resulting in the death of a Tibetan black bear (Ursus thibetanus) and several captive and wild bird species. We detected the virus in various tissues of the bear and a wild black-headed gull found dead in its enclosure using histopathology, two different in-situ detection techniques and next generation sequencing, all performed on formalin fixed paraffin embedded tissues. Phylogenetic analysis performed on HA gene segment showed that bear and gull strains shared 99.998% genetic identity, making the bird strain the closest related one. We detected the PB2 E627K mutation in minute quantities in the gull, whereas it predominated in the bear, which suggests that this mammalian adaptation marker was selected during the bear infection. Our results provide the first molecular and histopathological characterization of an H5N1 virus infection in this bear species.

microbiology↗

bioRxiv2024

Molecular detection using hybridization capture and next-generation sequencing reveals cross-species transmission of feline coronavirus type-1 between a domestic cat and a captive wild felid

Feline coronavirus (FCoV) infection normally causes mild or subclinical signs and is common in domestic cats. However, in some cats, FCoV infection can also lead to the development of feline infectious peritonitis (FIP)--a typically lethal disease. FCoV has two serotypes or genotypes, FCoV-1 and FCoV-2, both of which can cause FIP. The main difference between the genotypes is the viral spike (S) protein that determines tropism and pathogenicity, crucial mechanisms in the development of FIP. Subclinical infection and FIP have both been reported in wild felids, including in threatened species. Due to the high genetic variability of the S gene and the technical challenges to sequencing it, detection and characterization of FCoV in wild felids have mainly centered on other more conserved genes. Therefore, the genotype causing FIP in most wild felids remains unknown. Here, we report a retrospective molecular epidemiological investigation of FCoV in a zoological institution in the United States. In 2008, a domestic cat (Felis catus) and a Pallas cat (Otocolobus manul) sharing the same room succumbed to FIP. Using hybridization capture and next-generation sequencing, we detected and sequenced the complete 3-end of the viral genome ([~]8.2 Kb), including the S gene, from both felids. Our data show for the first time that FCoV-1 can be transmitted between domestic and wild felids and extends the known host range of FCoV-1. Our findings highlight the importance of identifying the genotype causing FIP, to develop effective control measures.

microbiology↗

bioRxiv2024

RESOLVING BIOLOGY'S DARK MATTER: SPECIES RICHNESS, SPATIOTEMPORAL DISTRIBUTION, AND COMMUNITY COMPOSITION OF A DARK TAXON

BackgroundZoologys dark matter comprises hyperdiverse, poorly known taxa that are numerically dominant but largely unstudied, even in temperate regions where charismatic taxa are well understood. It is everywhere, but high diversity, abundance, and small size have historically stymied its study. We demonstrate how entomological dark matter can be elucidated using high-throughput DNA barcoding ("megabarcoding"). We reveal the high abundance and diversity of scuttle flies (Diptera: Phoridae) in Sweden using 31,800 specimens from 37 sites across four seasonal periods. We investigate the number of scuttle fly species in Sweden and the environmental factors driving community changes across time and space. ResultsSwedish scuttle fly diversity is much higher than previously known, with 549 mOTUs (putative species) detected, compared to 374 previously recorded species. Hierarchical Modelling of Species Communities reveals that scuttle fly communities are highly structured by latitude and strongly driven by climatic factors. Large dissimilarities between sites and seasons are driven by turnover rather than nestedness. Climate changes are predicted to significantly affect the 47% of species that show significant responses to mean annual temperature. Results were robust whether using haplotype diversity or species-proxies (mOTUs) as response variables. Additionally, species-level models of common taxa adequately predict overall species richness. ConclusionsUnderstanding the bulk of the diversity around us is imperative during an era of biodiversity loss. We show that dark insect taxa can be efficiently characterized and surveyed with megabarcoding. Undersampling of rare taxa and choice of operational taxonomic units do not alter the main ecological inferences, making it an opportune time to tackle zoologys dark matter.

ecology↗

bioRxiv2024

SARS-CoV-2 outbreak in lions, tigers and hyenas at Denver Zoo

In late 2019, SARS-CoV-2 spilled-over from an animal host into humans, where it efficiently spread, resulting in the COVID-19 pandemic. Through both natural and experimental infections, we learned that many animal species are susceptible to SARS-CoV-2. Importantly, animals in close proximity to humans, including companion, farmed, and those at zoos and aquariums, became infected, and many studies demonstrated transmission to/from humans in these settings. In this study, we first review the literature of SARS-CoV-2 infections in tigers and lions, and compare species, sex, age, virus and antibody detection assay, and types, frequency and length of clinical signs, demonstrating broad heterogeneity amongst infections. We then describe a SARS-CoV-2 outbreak in lions, tigers and hyenas at Denver Zoo in late 2021. Animals were tested for viral RNA (vRNA) for four months. Lions had significantly more viral RNA in nasal swabs than both tigers and hyenas, and many individual lions experienced viral recrudescence after weeks of undetectable vRNA. Infectious virus was correlated with high levels of vRNA and was more likely to be detected earlier during infection. Four months post-infection, all tested animals generated robust neutralizing antibody titers. Animals were infected with Delta lineage AY.20 identical to a variant circulating at less than 1% in Colorado humans at that time, suggesting a single spillover event from an infected human spread within and between species housed at the zoo. Better understanding of epidemiology and susceptibility of SARS-CoV-2 infections in animals is critical to limit the current and future spread and protect animal and human health. ImportanceSurveillance and experimental testing have shown many animal species, including companion, wildlife, and conservatory, are susceptible to SARS-CoV-2. Early in the COVID-19 pandemic, big cats at zoological institutions were among the first documented cases of naturally infected animals; however, challenges in the ability to collect longitudinal samples in zoo animals have limited our understanding of SARS-CoV-2 kinetics and clearance in these settings. We measured SARS-CoV-2 infections over three months in lions, tigers and hyenas at Denver Zoo, and detected viral RNA, infectious virus, neutralizing antibodies, and recrudescence after initial clearance. We found lions had longer and higher levels of virus compared to the other species. All animals were infected by a rare viral lineage circulating in the human population, suggesting a single spillover followed by interspecies transmission. These data are important in better understanding natural SARS-CoV-2 spillover, spread and infection kinetics within multiple species of zoo animals.

microbiology↗

bioRxiv2024

Cross-species transmission of human hepatitis B virus to wild Neotropical primates

Hepatitis B virus (HBV) infects approximately one-third of the worlds human population and kills over one million people each year. HBV is also prevalent in Old World apes but not in New World primates. Human-to-primate transmission of HBV was suspected in zoo-captive monkeys and Mauritius macaques, but empirical data are scarce. Here, we collected blood and liver samples from 88 monkeys of 27 species in two areas of the Amazon, one with pristine forest and the other highly occupied and deforested by humans. A total of 17 (34.7%) out of 49 specimens from the human-occupied region tested positive for HBV. At this site, there was a positive relationship between human population density in the sampling location and the likelihood of primates being infected by HBV. Conversely, all 39 samples from the pristine forest tested negative for HBV. By sequencing a portion of the HBV S gene in five positive samples, each from a distinct primate genus, we found that four samples were closely related to the globally widespread human HBV-A strain, but not to the Americas-native HBV-F strain. The fifth sample aligned with the human HBV-D type, prevalent in the region where these samples were obtained. To our knowledge, this study represents the first reported cases of HBV in multiple wild New World primate species anywhere in the world. Our results suggest that primates were infected by strains brought into this part of Brazil by human immigrants, where HBV transmission may have been facilitated by the close contact between humans and monkeys due to high human occupation. This shows that the impact of human immigration, occupation and population growth in the Amazon extends beyond habitat loss; it also facilitates cross-species infections, potentially leading to the emergence of new, virulent viral strains that threaten both Amazonian biodiversity and human health. Author SummaryJean P. Boubli: Professor in Primate Ecology and Evolution, School of Science, Engineering and the Environment, University of Salford, Uk Hani R. El Bizri: PhD in Wildlife Conservation with a focus on Amazonian sustainable development. Analyst of Center for International Forestry Research (CIFOR), Bogor, Indonesia Luan F. Botelho-Souza: Masters and doctorate in experimental biology at the Universidade Federal de Rondonia/Brazil, Chrysoula Gubili: Researcher at the Fisheries Research Institute Nea Peramos, Kavala, Greece, specializing in population ecology and conservation genetics. Stephen J. Martin: Professor in Social Insects, School of Science, Engineering and the Environment, University of Salford, Uk Maisa da S. Araujo: Masters and doctorate in experimental biology, at the Universidade Federal de Rondonia/Brazil, Mariluce R. Messias: Professor of Zoology and curator of the Mammalogy Museum at the Universidade Federal de Rondonia/Brazil. Alcione de O. dos Santos: Masters and doctorate in experimental biology at the Universidade Federal de Rondonia/Brazil. Luiz S. Ozaki: Masters degree from Universidade de Brasilia and PhD in Physiological Sciences, Molecular Biology in Fukuoka, Kyushu, Japan. Andre V.C. Pereira: Bachelor in Biology the Universidade Federal de Rondonia/Brazil. Tony H. Katsuragawa: Masters and doctorate in experimental biology, at the Universidade Federal de Rondonia/Brazil. Ana Maisa Passos-Silva: Masters degree in experimental biology from the Federal University of Rondonia/Brazil. Luiz H. S. Gil: Masters in experimental biology, at the Universidade Federal de Rondonia/Brazil. Izeni P. Farias: Professor of Genetics, Universidade Federal do Amazonas, Brazil Juan M.V. Salcedo: Master in Tropical Medicine from the University of Brasilia/Brazil and PhD in Sciences from the University of Sao Paulo/Brazil. Tommy C. Burch: Masters in Biological Sciences and PhD candidate from the University of Salford, UK. Deusilene Vieira: Master in Tropical Medicine from the University of Brasilia/Brazil and PhD in Sciences from the University of Sao Paulo/Brazil.

pathology↗

bioRxiv2024

Emergence and Dynamics of Usutu and West Nile Viruses in the Netherlands, 2016-2022

BackgroundMosquito-borne arboviruses, including Usutu virus (USUV) and West Nile virus (WNV), are emerging threats in Europe, with changes in climate, land use, and increased travel and trade influencing their dynamics. Understanding the emergence and establishment of these viruses in new regions is critical for informing targeted mitigation of drivers of emergence and enhancing public and wildlife health preparedness. MethodsSeven years (2016-2022) of interconnected studies were conducted in the Netherlands. Live birds were sampled by volunteer bird ringers, dead birds were referred for sampling by citizen-scientists and zoological institutions, and mosquitoes were trapped. Samples were tested for USUV and WNV using RT-PCR and screened for Orthoflavivirus antibodies using protein microarray and neutralization assays. Sequencing and phylogenetic analyses were performed. FindingsUSUV was first detected in the Netherlands in 2016, caused large outbreaks in birds until 2018, and resurged in 2022. One dominant, enzootic lineage, co-circulated alongside several limited introductions of a second lineage. A localized WNV lineage 2 outbreak occurred in live birds and mosquitoes in 2020, with another positive bird in 2022 and serological evidence of continued circulation. InterpretationWe provide the first comprehensive, multi-year documentation of two emerging arboviruses in the Netherlands. USUV was enzootically maintained over seven years and was associated with substantial bird mortality. WNV is in an early stage of establishment with no bird mortality observed. Our integrated wildlife sampling was crucial in detecting a human WNV outbreak, bringing this emerging threat to public health attention. FundingZonMw, Dutch Research Council (NWO), European Union, Government of the Netherlands

microbiology↗

bioRxiv2025

Antero-posterior patterning in the brittle star Amphipholis squamata and the evolution of body plans across echinoderms

Although the adult pentaradial body plan of echinoderms evolved from a bilateral ancestor, identifying axial homologies between the morphologically divergent echinoderms and their bilaterian relatives has been an enduring problem in zoology. The expression of conserved bilaterian patterning genes in echinoderms provides a molecular framework for resolving this puzzle. Recent studies in juvenile asteroids suggest that the bilaterian antero-posterior axis maps onto the medio-lateral axis that is perpendicular to each of the five rays of the pentaradial body plan. Here we test this hypothesis in another echinoderm class, the ophiuroids, using the cosmopolitan brittle star Amphipholis squamata. Our results show that the general principles of axial patterning are similar to those described in asteroids, and comparisons with existing molecular data from other echinoderm taxa support the idea that medio-lateral deployment of the AP patterning program across the rays predates the evolution of the asterozoan and likely the echinoderm crown-groups. Our data also reveal expression differences between A. squamata and asteroids, which we attribute to secondary modifications specific to ophiuroids. Together, this work provides important comparative data to reconstruct the evolution of axial properties in echinoderm body plans.

evolutionary biology↗

bioRxiv2025

Impact of temperature on vector competence of Culex pipiens molestus: implications for Usutu virus transmission in temperate regions

IntroductionUsutu virus (USUV) has been detected annually in the southeast of England since 2020. USUV RNA has been identified in wild birds and mosquito populations, and exposure of captive birds to USUV at Zoological Society of London (ZSL) has also been confirmed. Since its first detection in London, USUVs distribution has expanded across the South East, highlighting a need to understand the transmission dynamics of this virus in the UK. The primary vectors of USUV in the UK are likely Culex pipiens mosquitoes. Two biotypes have been identified, the bird-feeding Cx. pipiens pipiens and Cx. pipiens molestus which shows no restriction in host preference. The latter may play an important role in transmitting USUV from birds to humans. MethodsA laboratory colony of Cx. pipiens molestus mosquitoes were orally infected with the London strain of USUV and, incubated at 22 {degrees}C, 20 {degrees}C and 18{degrees} C for up to 28 days. Body samples and mosquito saliva samples were collected and analysed using a quantitative real-time reverse transcription PCR to determine infection and transmission potential, respectively. ResultsUSUV RNA was detected in all sample times at all temperatures assessed; with the highest temperature (22 {degrees}C) showing the greatest proportion of saliva and body positive samples. At this temperature, there was also an eight-fold increase in the relative viral copy number in the mosquito bodies, which was unobserved at other experimental temperatures. When a more sensitive PCR assay was used at the lowest experimental temperature used (18 {degrees}C) USUV RNA was present in the mosquito saliva and body samples for longer and showed a greater proportion of positive samples when compared to 20 {degrees}C. ConclusionThis study has demonstrated that Cx. pipiens molestus may be able to transmit USUV at 22 {degrees}C. Active replication of USUV was identified in the mosquito bodies at 22 {degrees}C but could not be demonstrated at lower temperatures, suggesting that 20 {degrees}C to 22 {degrees}C may be an important temperature threshold in USUV replication and transmission. Utilisation of a more sensitive assay for the lower experimental temperatures revealed that USUV was detectable at 18 {degrees}C. Therefore, when conducting infection studies on temperate mosquito-borne viruses, it is important to consider assay sensitivity.

molecular biology↗

bioRxiv2025

Emergence Risks from Within? Metagenomic Analysis of Mosquito Viromes from Two Zoos Reveals Mosquito-associated Orthobunyaviruses in the UK

In regions naive to mosquito-borne threats, risk assessments often prioritise incursion events, particularly those driven by climate change and host movement. However, these same factors may also facilitate the emergence of viruses already circulating in local vector populations. The use of metagenomics to assess mosquito viromes has expanded in recent decades, enabling the detection of both arboviruses and insect-specific viruses (ISVs), and helping to fill gaps in our understanding of virus circulation. In particular, zoological collections are potential interfaces for virus transmission between mosquitoes, animals, and humans, making them valuable sentinel sites for surveillance. Although mosquito virome studies have been conducted globally, UK mosquito populations remain comparatively understudied. To address this gap, we investigated the viromes of two key mosquito species, Culex pipiens s.l. and Culiseta annulata, through metagenomic sequencing of specimens collected at two UK zoos between 2021 and 2022. A total of 4,042 mosquitoes underwent sequencing across 44 pools, leading to the identification of 26 viral genomes, including nine novel species spanning RNA and DNA virus families. Viral distribution patterns revealed viruses shared across all pools, supporting the possibility of a conserved virome between mosquito species. Notably, two novel Orthobunyavirus species (Family Peribunyaviridae), Atherstone and Deva viruses, were detected. These represent the first documented mosquito-associated orthobunyaviruses in the UK, a genus that includes several known pathogens. Classification using the machine-learning tool MosViR suggests Atherstone virus, present at both zoos, is a putative arbovirus warranting further investigation. Supporting this is the presence of a non-structural protein S (NSs), a known virulence factor in orthobunyaviruses. In contrast, Deva virus had no NSs ORF and was not predicted to be an arbovirus. These findings improve our understanding of UK mosquito viruses, indicate possible risks to people or animals from orthobunyaviruses, and highlight the role of metagenomic surveillance in detecting emerging viral threats.

genomics↗

bioRxiv2025

Seventy-five Years of Systematic Biology: Looking Back, Moving Forward

AbstractWhat does "systematic biology" mean today, where has it been in the past, and where is it going? We explore these questions by considering five elements - collaboration, integration, discourse, infrastructure, and society - that we think have allowed systematic biology to adapt to change and sustain growth without losing its unique identity. In the spirit of celebrating the 75th anniversary of our flagship journal, we generated a comprehensive dataset for all Systematic Biology and Systematic Zoology articles that we could locate (N = 5,150) and used bibliometric and textual analyses to illustrate ways in which our field has transformed over time. We offer our humble opinions on how our community can ensure that systematic biologists inherit an enlightening, dynamic, and enduring future.

evolutionary biology↗