Arbovirus and insect‐specific virus discovery in Kenya by novel six genera multiplex high‐resolution melting analysis

A broad diversity of arthropod‐borne viruses (arboviruses) of global health concern are endemic to East Africa, yet most surveillance efforts are limited to just a few key viral pathogens. Additionally, estimates of arbovirus diversity in the tropics are likely to be underestimated as their discovery has lagged significantly over past decades due to limitations in fast and sensitive arbovirus identification methods. Here, we developed a nearly pan‐arbovirus detection assay that uses high‐resolution melting (HRM) analysis of RT–PCR products from highly multiplexed assays to differentiate broad diversities of arboviruses. We differentiated 15 viral culture controls and seven additional synthetic viral DNA sequence controls, within Flavivirus, Alphavirus, Nairovirus, Phlebovirus, Orthobunyavirus and Thogotovirus genera. Among Bunyamwera, sindbis, dengue and Thogoto virus serial dilutions, detection by multiplex RT–PCR‐HRM was comparable to the gold standard Vero cell plaque assays. We applied our low‐cost method for enhanced broad‐range pathogen surveillance from mosquito samples collected in Kenya and identified diverse insect‐specific viruses, including a new clade in anopheline mosquitoes, and Wesselsbron virus, an arbovirus that can cause viral haemorrhagic fever in humans and has not previously been isolated in Kenya, in Culex spp. and Anopheles coustani mosquitoes. Our findings demonstrate how multiplex RT–PCR‐HRM can identify novel viral diversities and potential disease threats that may not be included in pathogen detection panels of routine surveillance efforts. This approach can be adapted to other pathogens to enhance disease surveillance and pathogen discovery efforts, as well as the study of pathogen diversity and viral evolutionary ecology.     

A new hero emerges: another exceptional mammalian spine and its potential adaptive significance

The hero shrew''s (Scutisorex somereni) massive interlocking lumbar vertebrae represent the most extreme modification of the vertebral column known in mammals. No intermediate form of this remarkable morphology is known, nor is there any convincing theory to explain its functional significance. We document a new species in the heretofore monotypic genus Scutisorex; the new species possesses cranial and vertebral features representing intermediate character states between S. somereni and other shrews. Phylogenetic analyses of DNA sequences support a sister relationship between the new species and S. somereni. While the function of the unusual spine in Scutisorex is unknown, it gives these small animals incredible vertebral strength. Based on field observations, we hypothesize that the unique vertebral column is an adaptation allowing these shrews to lever heavy or compressive objects to access concentrated food resources inaccessible to other animals.     

Metagenomic Survey for Viruses in Western Arctic Caribou,Alaska, through Iterative Assembly of Taxonomic Units

Pathogen surveillance in animals does not provide a sufficient level of vigilance because it is generally confined to surveillance of pathogens with known economic impact in domestic animals and practically nonexistent in wildlife species. As most (re-)emerging viral infections originate from animal sources, it is important to obtain insight into viral pathogens present in the wildlife reservoir from a public health perspective. When monitoring living, free-ranging wildlife for viruses, sample collection can be challenging and availability of nucleic acids isolated from samples is often limited. The development of viral metagenomics platforms allows a more comprehensive inventory of viruses present in wildlife. We report a metagenomic viral survey of the Western Arctic herd of barren ground caribou (Rangifer tarandus granti) in Alaska, USA. The presence of mammalian viruses in eye and nose swabs of 39 free-ranging caribou was investigated by random amplification combined with a metagenomic analysis approach that applied exhaustive iterative assembly of sequencing results to define taxonomic units of each metagenome. Through homology search methods we identified the presence of several mammalian viruses, including different papillomaviruses, a novel parvovirus, polyomavirus, and a virus that potentially represents a member of a novel genus in the family Coronaviridae.     

Radiotelemetry Applied to Field Studies of Shrews

Abstract: Radiotelemetry is a widely used method to study ecology, behavior, and physiology of different animals but has rarely been used on shrews. Small body size, wide neck and narrow skull, high mobility, and fragility of shrews cause problems for both transmitter attachment and the safety of the animals. We developed a method for nonpermanent attachment of transmitters, which allowed us to track such small mammals as the Eurasian water shrews (Neomys fodiens; n = 39, mean body mass 14.9 g), Mediterranean water shrews (Neomys anomalus; n = 32, 10.9 g), and common shrews (Sorex araneus; n = 51, 8.1 g). We used microtransmitters weighing 0.47 g, but those we applied to the larger Neomys species were heavier (0.67 g) because we fortified them with a layer of hard material to prevent damage from biting. We glued a transmitter directly to the skin on a shrew's back, with the anterior edges particularly well sealed. We tracked shrews in the wild and in outdoor enclosures. Transmitters usually dropped off together with peeled skin (on average, after 56.0 hr, n = 92 observations), but if not dropped and if the signal was not lost, mean duration of monitoring was 96.7 hours (n = 37) and in 2 cases exceeded 194 hours. Other advantages of our attachment method were 1) it was less invasive and easier to apply than implantation of transmitters into the body cavity, and 2) we could find dropped transmitters and reuse them. We give suggestions on how to minimize the risk of injury to animals by correct handling, manipulation, and gluing. In conclusion, we recommend radiotelemetry as a useful technique for studying shrew behavior in both free-living populations and experimental enclosures.     

Bergmann's rule in shrews: geographical variation of body size in Palearctic Sorex species

According to Bergmann's rule, individuals of a given species tend to be larger in colder (northern) climates. Traditional explanation points to the relatively lower surface‐to‐volume ratio in larger animals and, consequently, relatively lower costs of thermoregulation. We examined intraspecific covariation of body size with geographical location and climate in five species of Sorex shrews, animals that are among the smallest extant mammals. The condylobasal length of skull (CBL), compiled from literature data and measured on museum specimens, was used as an indicator of the overall body size of shrews. Surprisingly, in three out of five shrew species, the CBL was negatively correlated with latitude, and the same trend, although not statistically significant, was found in the fourth species. In general, shrews were smaller in colder areas, as evidenced by the positive correlations between the CBL and temperature. In two species, these positive correlations appeared when the effect of longitude was held constant in the partial correlation analysis. Characteristically, the strongest negative correlation with latitude and positive with temperatures was found in S. minutus, the smallest species under study. Shrews were in general larger in environments with high actual evapotranspiration. Body mass reviewed in S. araneus paralleled the pattern found in the CBL variation in this species, i.e. it decreased northward, both in summer‐ and winter‐caught animals. In addition, contrary to the widely accepted ? but not rigorously tested ? belief, body mass recession from summer to winter (the Dehnel Effect) did not correlate with latitude. We concluded that shrews followed the converse to Bergmann's rule, and hypothesize that part of their body size variation along the west‐east axis may be explained by character displacement. We also hypothesize that scarcity of food, especially in winter, is a major factor selecting for small body size in shrews in northern areas, as smaller individuals should require less food. © 2003 The Linnean Society of London, Biological Journal of the Linnean Society, 2003, 78 , 365–381.     

First results on the feeding ecology of sympatric shrews (Insectivora: Soricidae) in the Tai National Park,Ivory Coast

The feeding ecology of a multi-species community of shrews inhabiting secondary forest and cacao-coffee plantations in the Tai National Park (Ivory Coast) was investigated. A total of 553 shrews were captured and 194 alimentary tracts were examined. Ten species were found, includingSylvisorex megalura and nine species ofCrocidura, forming a series with respect to body size. New ecological data on these little known African species are presented. All species of shrews ate a wide diversity of arthropods, with Coleoptera, Araneae, Formicidae and Diplopoda making the largest dietary contributions. Lumbricidae were eaten by two species.C. obscurior had an exceptionally long intestine for its size but there was no evidence of dietary specialisation in this or other shrew species. All species investigated ate predominantly small prey and there was no correlation between size of prey items consumed and body mass of shrew species. There was little evidence of resource partitioning amongst the shrews, despite differences in body size.     

Small mammal abundance in relation to microhabitat in a dry sub-humid grassland in Kenya

Six species of rodents and two species of insectivores, live-trapped on a 1 ha grid over a 27 month period, were studied in relation to microhabitat factors in a dry sub-humid grassland in Kenya. Wet season peaks of small mammal species were followed by low numbers trapped during the dry seasons. Species distributions were correlated with different microhabitat parameters, and the separation of species with apparently similar environmental requirements could be explained largely in terms of current knowledge of the biology of the species. Correlation analyses indicated an ecological separation of two closely-related Mus species and of two Crocidura shrews. Overlap indices were extremely high, ranging from 0.556 to 0.877 for all combinations of pairs of species. It seems likely that neither food nor micro-habitat partitioning completely explains the coexistence of the small mammal species and it is suggested that populations are highly transient, moving from refuge areas into temporarily favourable areas during the wet season.     

The isolation of histoplasma duboisii and keratinophilic fungi from soils of east africa

Summary Isolation ofHistoplasma duboisii marks the first incidence of this African pathogen being found in the soil of Kenya.Investigation of keratinophilic fungi in soil from Kenya and Tanzania produced 27 isolates of dermatophytes belonging to the four known genera of this group.Trichophyton evolceanui was found in five of these samples.It is to be noted that localities frequented by wild and domestic animals produced a higher incidence of keratinophilic fungi than did the unshady, isolated areas.     

Molecular phylogeny of Armillaria from the Patagonian Andes

A number of species in the plant pathogen genus Armillaria are known from South America where they cause root rot disease on a wide variety of hosts. Knowledge pertaining to phylogenetic relationships of these species with those of other Armillaria species is almost non-existent. In addition, very few cultures representing these species are available, making DNA-based phylogenetic analyses impossible. The aim of this study was to characterise a collection of Armillaria isolates from the Patagonian Andes using DNA sequences and to determine their phylogenetic relationships with other Armillaria species. DNA sequences were obtained from the internal transcribed regions (ITS1, 5.8S and ITS4) and ribosomal large subunit (LSU) gene and used in phylogenetic analyses. Phylogenetic trees generated from the sequences separated the Armillaria isolates into four lineages. Lineages I and II represented A. novae-zelandiae and A. luteobubalina, respectively. Isolates belonging to A. novae-zelandiae from Malaysia, New Zealand, Australia and South America showed considerable intra-clade sub-structure. Lineages III and IV are probably distinct species and are most closely related to A. hinnulea and an unnamed species isolated from New Zealand and Kenya. This is the first comprehensive study of the phylogenetic relationships of Armillaria species from Patagonia and it provides a foundation for future research in this region.     

Unexpectedly high diversity of trypanosomes in small sub-Saharan mammals

The extremely species-rich genus Trypanosoma has recently been divided into 16 subgenera, most of which show fairly high host specificity, including the subgenus Herpetosoma parasitizing mainly rodents. Although most Herpetosoma spp. are highly host-specific, the best-known representative, Trypanosoma lewisi, has a cosmopolitan distribution and low host specificity. The present study investigates the general diversity of small mammal trypanosomes in East and Central Africa and the penetration of invasive T. lewisi into communities of native rodents. An extensive study of blood and tissue samples from Afrotropical micromammals (1528 rodents, 135 shrews, and five sengis belonging to 37 genera and 133 species) captured in the Central African Republic, Ethiopia, Kenya, Malawi, Mozambique, Tanzania, and Zambia revealed 187 (11.2%) trypanosome-positive individuals. The prevalence of trypanosomes in host genera ranged from 2.1% in Aethomys to 37.1% in Lemniscomys. The only previously known trypanosome detected in our dataset was T. lewisi, newly found in Ethiopia, Kenya, and Tanzania in a wide range of native rodent hosts. Besides T. lewisi, 18S rRNA sequencing revealed 48 additional unique Herpetosoma genotypes representing at least 15 putative new species, which doubles the known sequence-based diversity of this subgenus, and approaches the true species richness in the study area. The other two genotypes represent two new species belonging to the subgenera Ornithotrypanum and Squamatrypanum. The trypanosomes of white-toothed shrews (Crocidura spp.) form a new phylogroup of Herpetosoma, unrelated to flagellates previously detected in insectivores. With 13 documented species, Ethiopia was the richest region for trypanosome diversity, which corresponds to the very diverse environments and generally high biodiversity of this country. We conclude that besides T. lewisi, the subgenus Herpetosoma is highly host-specific (e.g., species parasitizing the rodent genera Acomys and Gerbilliscus). Furthermore, several newly detected trypanosome species are specific to their endemic hosts, such as brush-furred mice (Lophuromys), dormice (Graphiurus), and white-toothed shrews (Crocidura).     

Distribution of small rodents and shrews in a lowland rain forest zone of Nigeria, with observations on their reproductive biology

This paper describes the habitat preferences of small rodents and shrews from April 1984 to March 1985 in four types of vegetation in a lowland rain forest zone of Nigeria. The vegetation types were high forest, scrub, farmlands and teak plantations in Ogba Forest Reserve, Nigeria. A collection of 359 small mammals, consisting of six species of small rodents and four species of shrews, was made in the study area. The small rodents and their percentage composition were Mus musculoides (26.2%), Praomys tullbergi (16.2%), Mastomys natalensis (7.0%), Lophuromys sikapusi (4.5%), Lemniscomys striatus (2.5%), and Arvicanthis niloticus (1.1%). The shrews were Crocidura nigeriae (19.5%), C. crossei (12.3%), C.grandiceps (7.8%) and C. ftavescens manni (3.1%). The vegetational distribution of the small rodents and shrews is discussed. In the high forest, seven species of small mammals were caught; these were Crocidura nigeriae, C. crossei, C.grandiceps, C.flavescens manni, Mus musculoides, Praomys tullbergi and Lophuromys sikapusi. Mus musculoides, Crocidura nigeriae and Praomys tullbergi were more abundant in the high forest. All the species of small mammals recorded in the study area, except Arvicanthis niloticus, were found in the scrub. In the farmlands, all ten species of small mammals were trapped. Mus musculoides was the commonest species in the farmlands. Three species of shrews, Crocidura crossei, C. grandiceps and C.flavescens manni, and also two species of small rodents, Mus musculoides and Mastomys natalensis were caught in the teak plantation. The reproductive data for Mus musculoides, Crocidura nigeriae and Praomys tullbergi show that breeding in these species occurs throughout most months of the year. Fecundity appears to be maximal in the wet season in Mus musculoides and Crocidura nigeriae and in the dry season in the case of Praomys tullbergi.     

Genetic identity of the critically endangered Wimmer's shrew Crocidura wimmeri

Coastal primary rainforests have suffered damage in Côte d'Ivoire as a result of a lack of protection and urban pressures. Consequently, the highly endemic and critically endangered Wimmer's shrew, Crocidura wimmeri, known only from its type locality, Adiopodoumé, near Abidjan, was considered to have been extinct since 1976. Shrew species assignment is often problematic because of strong phenotypic similarities among many species. The phylogenetic position of C. wimmeri within the African Crocidura species should thus be clarified. In light of its recent rediscovery in the nearby small Banco National Park (34 km²), we investigated the genetic identity of seven specimens of C. wimmeri, based on 1020 bp of the mitochondrial DNA cytochrome b gene compared to other species sampled in the same region and published sequences from GenBank. Crocidura wimmeri formed a well‐defined clade, the closest‐related species being Crocidura sp., with a distance of 9.3%, a yet unknown species from Taï and Ziama forests. These results thus confirmed the validity of this species. This genetic characterization not only contributes to our knowledge of the evolution of West African shrews, but also may help in the discovery of additional populations of this critically endangered species. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 224–229.     

Behavioural mechanisms of conflict avoidance among shrews

Stable co-existence of similar species should be facilitated by mechanisms impairing, besides exploitative, interference competition. We investigated avoidance of intra- and interspecific conflicts in a four-species community of shrews [Sorex minutus Linnaeus, 1766,S. araneus Linnaeus, 1758,Neomys anomalus Cabrera, 1907, andN. fodiens (Pennant, 1771)], using the method of dyadic encounters in a neutral arena. We tested whether the use of passive (habituation, reduction of mobility, increase of inter-individual distance, and stillness) and active (‘to-and-fro’ and ‘keeping distance’ behaviours) forms of conflict avoidance depends on species, size or domination rank. The duration of conflicts was positively correlated with mobility and negatively with inter-individual distance, whereas it was unrelated to time of stillness and the active forms. The repertoire of conflict avoidance mechanisms was not species-specific and the display of these mechanisms depended rather on the size and domination rank of animals participating in a given interaction. In contrast to rodents, shrews did not avoid conflicts by the most passive forms: freeze and stillness reactions. All other forms were used with a higher or lower efficiency by all species. However, consistent with our predictions, large shrews (asN. fodiens) used mainly the passive mechanisms of conflicts avoidance (‘wait-and-see’ strategy), whereas small shrews (asS. minutus) invest proportionally more time in active forms (‘escape’ strategy).     

Incidence and pathology of common viruses of nerine in Tasmania

This is the first study of the incidence of common viruses within bulb stocks of nerine in Australia. Five viruses were frequently found within commercial stocks but were never associated with obvious viral symptom expression in the field. This study found cucumber mosaic virus (CMV) infections of Nerine flexuosa, and nerine virus Y (NeVY) infections of N. flexuosa, N. bowdenii and N. fothergilli which we believe to be new host records. Virus incidence varied with virus species, host species, and sample site but did not vary within the same bulb stocks over a 2 yr period. Incidence of the major viruses appeared to correlate with the efficiency of transfer from mother bulb to offsets, suggesting that virus status in Tasmania may be largely determined by the incidence of viruses within original planting material and by subsequent vegetative spread. No effect of the most frequent virus infections was found on the efficiency of flowering, flower quality, and dormant bulb fresh weight in N. flexuosa and N. fothergilli with the exception of a small (16.8%), but economically unimportant decrease in scape length of N. flexuosa co‐infected with NeLV and NeVY. It is concluded that current virus infections in nerine in Tasmania pose little commercial significance to the developing cut flower industry, however discovery of NeYSV in one sample of Lycoris aurea warrants monitoring to ensure that spread of this important pathogen does not occur.     

Carving out turf in a biodiversity hotspot: multiple,previously unrecognized shrew species co‐occur on Java Island,Indonesia

In theory, competition among species in a shared habitat results in niche separation. In the case of small recondite mammals such as shrews, little is known about their autecologies, leaving open questions regarding the degree to which closely related species co‐occur and how or whether ecological niches are partitioned. The extent to which species are able to coexist may depend on the degree to which they exploit different features of their habitat, which may in turn influence our ability to recognize them as species. We explored these issues in a biodiversity hotspot, by surveying shrew (genus Crocidura) diversity on the Indonesian island of Java. We sequenced portions of nine unlinked genes in 100–117 specimens of Javan shrews and incorporated homologous data from most known Crocidura species from other parts of island South‐East Asia. Current taxonomy recognizes four Crocidura species on Java, including two endemics. However, our phylogenetic, population genetic and species delimitation analyses identify five species on the island, and all are endemic to Java. While the individual ranges of these species may not overlap in their entirety, we found up to four species living syntopically and all five species co‐occurring on one mountain. Differences in species' body size, use of above ground‐level habitats by one species and habitat partitioning along ecological gradients may have facilitated species diversification and coexistence.     

Small mammal mycophagy in hemiboreal forest communities of Lithuania

The diets of small mammals in different hemiboreal spruce-dominated, oak-dominated and mixed forests in western part of Lithuania were studied by examination of fungal spores in fresh fecal pellets of caught animals. In the diets of mice (Apodemus spp.), bank voles (Myodes glareolus), and common and pygmy shrews (Sorex araneus and S. minutus), 22 different fungal taxa were identified, 15 of which were hypogeous fungi. The sporocarp abundance and the spores in fecal samples of Elaphomyces fungi prevailed in study area during this investigation. Although most of the captured individuals consumed fungi, the consumption varied among small mammal species. The data show that the fungi were more frequent and taxonomically diverse in Myodes glareolus than in Apodemus spp. diets. The study provided evidence that the fungal component in the diets of insectivorous Sorex species is more diverse than previously known. The availability of sporocarps and the fungal component in the diets of small mammals showed seasonal effects. Annual hypogeous and epigeous sporocarp abundances did not vary significantly across forest types. The significant difference in mycophagy was observed across all forest cover types, with the greatest fungal diversity in fecal samples collected in mixed coniferous-deciduous tree stands.     

The feeding ecology of the European Water shrew

This review examines our knowledge of the feeding ecology of the European Water shrew, Neomys fodiens. It describes and discusses the different methods which have been used to study the diets of Water shrews, from the first casual observations to the discovery of food caches and food remains in the wild, and the analysis of stomach contents and faecal pellets of trapped animals. These studies have revealed that Water shrews feed on a variety of invertebrate and vertebrate prey from tiny freshwater dip-teran larvae to sizable fish and frogs. Although aquatic prey are seen to be the major dietary component in most studies, terrestrial prey are also important and may be eaten in an equal proportion. Laboratory studies have increased our knowledge of the foraging behaviour of Water shrews but there is still much to be learnt, especially about underwater prey location.     

Viral gut metagenomics of sympatric wild and domestic canids,and monitoring of viruses: Insights from an endangered wolf population

Animal host–microbe interactions are a relevant concern for wildlife conservation, particularly regarding generalist pathogens, where domestic host species can play a role in the transmission of infectious agents, such as viruses, to wild animals. Knowledge on viral circulation in wild host species is still scarce and can be improved by the recent advent of modern molecular approaches. We aimed to characterize the fecal virome and identify viruses of potential conservation relevance of diarrheic free‐ranging wolves and sympatric domestic dogs from Central Portugal, where a small and threatened wolf population persists in a highly anthropogenically modified landscape. Using viral metagenomics, we screened diarrheic stools collected from wolves (n = 8), feral dogs (n = 4), and pet dogs (n = 6), all collected within wolf range. We detected novel highly divergent viruses as well as known viral pathogens with established effects on population dynamics, including canine distemper virus, a novel bocavirus, and canine minute virus. Furthermore, we performed a 4‐year survey for the six wolf packs comprising this endangered wolf population, screening 93 fecal samples from 36 genetically identified wolves for canine distemper virus and the novel bocavirus, previously identified using our metagenomics approach. Our novel approach using metagenomics for viral screening in noninvasive samples of wolves and dogs has profound implications on the knowledge of both virology and wildlife diseases, establishing a complementary tool to traditional screening methods for the conservation of threatened species.     

Shedding of Infectious Borna Disease Virus-1 in Living Bicolored White-Toothed Shrews

Background

Many RNA viruses arise from animal reservoirs, namely bats, rodents and insectivores but mechanisms of virus maintenance and transmission still need to be addressed. The bicolored white-toothed shrew (Crocidura leucodon) has recently been identified as reservoir of the neurotropic Borna disease virus 1 (BoDV-1).

Principal Findings

Six out of eleven wild living bicoloured white-toothed shrews were trapped and revealed to be naturally infected with BoDV-1. All shrews were monitored in captivity in a long-term study over a time period up to 600 days that differed between the individual shrews. Interestingly, all six animals showed an asymptomatic course of infection despite virus shedding via various routes indicating a highly adapted host-pathogen interaction. Infectious virus and viral RNA were demonstrated in saliva, urine, skin swabs, lacrimal fluid and faeces, both during the first 8 weeks of the investigation period and for long time shedding after more than 250 days in captivity.

Conclusions

The various ways of shedding ensure successful virus maintenance in the reservoir population but also transmission to accidental hosts such as horses and sheep. Naturally BoDV-1-infected living shrews serve as excellent tool to unravel host and pathogen factors responsible for persistent viral co-existence in reservoir species while maintaining their physiological integrity despite high viral load in many organ systems.     

Novel Polyomaviruses of Nonhuman Primates: Genetic and Serological Predictors for the Existence of Multiple Unknown Polyomaviruses within the Human Population

Polyomaviruses are a family of small non-enveloped DNA viruses that encode oncogenes and have been associated, to greater or lesser extent, with human disease and cancer. Currently, twelve polyomaviruses are known to circulate within the human population. To further examine the diversity of human polyomaviruses, we have utilized a combinatorial approach comprised of initial degenerate primer-based PCR identification and phylogenetic analysis of nonhuman primate (NHP) polyomavirus species, followed by polyomavirus-specific serological analysis of human sera. Using this approach we identified twenty novel NHP polyomaviruses: nine in great apes (six in chimpanzees, two in gorillas and one in orangutan), five in Old World monkeys and six in New World monkeys. Phylogenetic analysis indicated that only four of the nine chimpanzee polyomaviruses (six novel and three previously identified) had known close human counterparts. To determine whether the remaining chimpanzee polyomaviruses had potential human counterparts, the major viral capsid proteins (VP1) of four chimpanzee polyomaviruses were expressed in E. coli for use as antigens in enzyme-linked immunoassay (ELISA). Human serum/plasma samples from both Côte d''Ivoire and Germany showed frequent seropositivity for the four viruses. Antibody pre-adsorption-based ELISA excluded the possibility that reactivities resulted from binding to known human polyomaviruses. Together, these results support the existence of additional polyomaviruses circulating within the human population that are genetically and serologically related to existing chimpanzee polyomaviruses.