Journal of General Virology (2009), 90, 2015–2022 DOI 10.1099/vir.0.

010694-0

Detection of polyoma and corona viruses in bats of

Canada
Vikram Misra,1 Timothy Dumonceaux,2 Jack Dubois,3 Craig Willis,4
Susan Nadin-Davis,5 Alberto Severini,2 Alex Wandeler,5 Robbin Lindsay2
and Harvey Artsob2
Correspondence 1
Department of Veterinary Microbiology, Western College of Veterinary Medicine, 52 Campus Road,
Vikram Misra University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
vikram.misra@usask.ca 2
National Microbiology Laboratory, Canadian Science Centre for Human and Animal Disease,
1015 Arlington Street, Winnipeg, MB R3E 3R2, Canada
3
Manitoba Conservation, Wildlife and Ecosystem Protection Branch, Box 24,
200 Saulteaux Crescent, Winnipeg, MB R3J 3W3, Canada
4
Department of Biology and Centre for Forest Interdisciplinary Research, University of Winnipeg,
515 Portage Avenue, Winnipeg, MB R3B 2E9, Canada
5
Centre of Expertise for Rabies, Ottawa Laboratory-Fallowfield, Canadian Food Inspection Agency,
3851 Fallowfield Road, Ottawa, ON K2H 8P9, Canada

Several instances of emerging diseases in humans appear to be caused by the spillover of viruses
endemic to bats, either directly or through other animal intermediaries. The objective of this study
was to detect, identify and characterize viruses in bats in the province of Manitoba and other
regions of Canada. Bats were sampled from three sources: live-trapped Myotis lucifugus from
Manitoba, rabies-negative Eptesicus fuscus, M. lucifugus, M. yumanensis, M. septentrionalis, M.
californicus, M. evotis, Lasionycteris (L.) noctivagans and Lasiurus (Las.) cinereus, provided by
the Centre of Expertise for Rabies of the Canadian Food Inspection Agency (CFIA), and L.
noctivagans, Las. cinereus and Las. borealis collected from a wind farm in Manitoba. We
attempted to isolate viruses from fresh tissue samples taken from trapped bats in cultured cells of
bat, primate, rodent, porcine, ovine and avian origin. We also screened bat tissues by PCR using
primers designed to amplify nucleic acids from members of certain families of viruses. We
detected RNA of a group 1 coronavirus from M. lucifugus (3 of 31 animals) and DNA from an as-
Received 29 January 2009 yet undescribed polyomavirus from female M. lucifugus (4 of 31 animals) and M. californicus
Accepted 31 March 2009 (pooled tissues from two females).

INTRODUCTION last two decades are enzootic in bats and are transmitted,
albeit infrequently, to humans and domestic animals
In a recent article, Jones et al. (2008) examined over 330
(reviewed by Calisher et al., 2006; Dobson, 2005; Wong
events of emerging infectious disease in humans over a
et al., 2007). Economic or sociological conditions that lead
60 year period from 1940 to 2004. They concluded that
to an increase in bat–human contact appear to predispose
most of the events were zoonotic and that over 70 % of
these cross-species transmissions. In some of the emergent
these could be attributed to wildlife. Other studies (Taylor
viruses, such as Hendra and Nipah viruses, there is
et al., 2001; Woolhouse & Gowtage-Sequeria, 2005)
evidence of direct transmission of bat viruses to horses
provide similar estimates. Several of the zoonotic viruses
and swine and from them to humans. Nipah virus has also
that have emerged in various regions of the world in the
been transmitted directly from bats to humans (Luby et al.,
2006) and between humans (Gurley et al., 2007). For other
The GenBank/EMBL/DDBJ accession number of the complete
nucleotide sequence of the Myotis polyomavirus and deduced amino
viruses such as severe acute respiratory syndrome (SARS)
acid sequence is FJ188392. (Lau et al., 2005), Marburg (Towner et al., 2007) and Ebola
(Leroy et al., 2005), viruses similar to those causing disease
Three supplementary tables showing primer sequences, background
information on bat specimens and GenBank accession numbers of the in people have been isolated from or detected in bats.
sequences analysed in this study are available with the online version of Melaka, a virus similar to Tioman virus previously isolated
this paper. from bats (Chua et al., 2007), was recovered from an

010694 G 2009 SGM Printed in Great Britain 2015

V. Misra and others

individual with severe respiratory disease. In addition, bat bats) have rarely been encountered by people historically,
lyssaviruses in the UK (Nathwani et al., 2003) and Australia because they roost exclusively in the foliage of trees (Willis
(Hanna et al., 2000; Samaratunga et al., 1998), both of & Brigham, 2005). The third migratory species (the silver-
which are considered to be free of terrestrial rabies, have haired bat) is also considered a tree-roosting species (Betts,
caused fatal disease in people. 1998) but has been more often encountered by people
when roosting in deadfall or woodpiles
Increased human development in wilderness areas of
Canada and encroachment into bat habitats could lead to If bats indeed harbour viruses that have the potential to
similar inter-species transmission of potentially zoonotic cause severe disease in humans and other domestic
viruses that may be present in bats, particularly via species animals, and if climatic and socio-economic changes are
which can roost in human-made structures. Additionally, likely to lead to conditions that facilitate contact between
widespread mortality of the so-called ‘migratory tree bats’, bats and domestic animals, it might be useful to identify
that has now been documented at industrial-scale wind viruses that infect bats. Such surveys have been conducted
energy facilities throughout North America (Arnett et al., elsewhere in the world with the identification of several
2008; Baerwald et al., 2008; Betts, 1998) has created the previously unknown bat viruses. Because of the devastating
potential for direct virus transmission between formerly effect of SARS on the economy of several countries, most of
cryptic forest bat species and domestic pets or livestock and these studies have specifically targeted coronaviruses
indirect transmission to wildlife species which scavenge bat (Carrington et al., 2008; Chu et al., 2006, Dominguez et
carcasses at wind turbines (e.g. foxes, skunks and crows; al., 2007; Gloza-Rausch et al., 2008; Lau et al., 2005; Poon
Klug & Barclay, 2008). Identification of such viruses and et al., 2005). Our objective in this study was to carry out a
characterization of their biology and epizootiology would preliminary screen of bats in Canada for the presence of
be invaluable in anticipating and possibly preventing viruses of several families. We attempted to isolate viruses
transmission to humans or, if it occurred, in controlling from fresh tissue samples taken from trapped bats in
its spread and impact. cultured cells of bat, primate, rodent, porcine, ovine and
At least 17 species of bats have been detected in Canada. avian origin. We also screened nucleic acids purified from
These include little brown bat (Myotis lucifugus), Yuma these samples by PCR using low-specificity primers
myotis (M. yumanensis), long-legged myotis (M. volans), designed to amplify nucleic acids from all members of
fringed myotis (M. thysanodes), northern myotis (M. certain families of viruses. We detected RNA related to that
septentrionalis), eastern small-footed myotis (M. leibii), of Rocky Mountain bat coronavirus (RMBCV) from little
Keen’s myotis (M. keenii), long-eared myotis (M. evotis), brown bats and DNA of a previously undescribed
western small-footed myotis (M. ciliolabrum), California polyomavirus from female little brown bats and
myotis (M. californicus), big brown bat (Eptesicus fuscus), California myotis.
silver-haired bat [Lasionycteris (L.) noctivagans], hoary bat
[Lasiurus (Las.) cinereus], red bat (Las. borealis), tri-
coloured bat (formerly the eastern pipstrelle, Perimyotis METHODS
subflavus), spotted bat (Euderma maculatum) and Bats. Bats for the study were obtained from three sources.
Townsend’s big-eared bat (Corynorhinus townsendii). Of
these species, the little brown bat, silver-haired bat, hoary (i) Little brown bats were trapped using a harp trap set up at a cave in
northern Manitoba during a September 2007 mating swarm. Some
bat, northern myotis, big brown bat and eastern red bat hibernating bats were also collected from hibernacula in two other
have been observed in the province of Manitoba (details in caves in May 2008. Bats were euthanized on site by an overdose of
Bilecki, 2003). During winter in Manitoba, the little and inhaled isoflurane (Aerrane) and placed on ice for transport to the
big brown bats and northern myotis bats hibernate, while National Microbiology Laboratory (NML) in Winnipeg. Bats were
the three other local species, the ‘migratory tree bats’ processed for virus isolation within 48 h of collection. A Wildlife
(silver-haired bat, hoary bat and eastern red bat), migrate Scientific Permit (WB 06638) to trap the bats was obtained from the
south. The little brown and northern myotis bats hibernate Wildlife and Ecosystem Protection Branch of Manitoba Conservation.
The bats were treated in accordance with procedures approved by the
in caves in the extensive limestone karst in the inter-lake
Animal Care Committee of the Canadian Science Centre for Human
and northern regions of Manitoba. Big brown bat and Animal Health (H-07-001 Rev.1). All individuals handling bats
hibernacula have not as-yet been identified. The little had been immunized against rabies and, according to recent tests, had
brown bats congregate at the caves in mid-August to late antibody levels considered protective.
September, presumably to mate. Later in the year, they
(ii) The Centre of Expertise for Rabies at the Canadian Food
move to the caves where they habitually hibernate during Inspection Agency (CFIA) provided bats diagnosed as negative for
winter. The bats leave the caves in mid-May to early June rabies. These bats had been submitted to the CFIA laboratories in
and females form maternity colonies in tree hollows and Lethbridge, Alberta and Ottawa, Ontario, and frozen at 220 uC after
other structures, including buildings. Little brown bats, in diagnosis. This group of bats included little and big brown bats,
particular, are common inhabitants of buildings through- silver-haired bats, hoary bats, Yuma myotis and California myotis.
out their range and are, therefore, the species most likely to (iii) Dead bats were collected for a survey of bat mortality at a
be encountered by humans in Manitoba. In contrast, two Manitoba wind farm. These included silver-haired, hoary and red
of the three migratory tree bats (hoary and eastern red bats.

2016 Journal of General Virology 90

 Viruses in bats of Canada

Sampling tissues. All bats were processed in biosecurity level 3 MS2 RNA, seeded into raw samples as a control for RNA extraction,
facilities at either the NML or the CFIA Fallowfield laboratory in was detected using quantitative RT-PCR (Dreier et al., 2005) by the
Ottawa. Carcasses were sprayed with 70 % ethanol and placed on virology facility at the NML.
absorbent paper for 5 min before dissection. One kidney and spleen,
Broad spectrum PCR for polyomavirus detected a conserved portion
and portions of liver, lung, small intestine and brain were removed
of the VP1 and VP3 genes (Johne et al., 2005). The remainder of the
from each bat. All tissues from each live-trapped bat were pooled. For
genome of polyomaviruses from three bats was obtained by ‘long-
bats obtained from CFIA and the wind farm, tissues from up to four
range’ PCR using primers facing outwards from the initial product of
bats of the same species were pooled. BA [Medium 199 (Gibco), 50 mM
the VP1 gene. The larger PCR product was cloned into pCRTOPO
Tris/HCl, pH 7.6, 1 % BSA and penicillin–streptomycin (Gibco)] was
using a PCR cloning kit (Invitrogen). The sequence of the fragment
added to tissues from individual animals (2 ml) and pooled tissues was determined using primers bracketing the cloning site of
(4 ml) and the tissues were homogenized either in a Polytron pCRTOPO and additional primers were designed as sequencing
homogenizer or by shaking with a sterile ball bearing in a Retsch progressed. Sequences were aligned into contigs using DNASTAR and
M300 oscillating homogenizer. The samples were then centrifuged at represented complete sequences of both DNA strands. Open reading
16 000 g for 5 min and the supernatant was transferred to a fresh tube. frames (ORFs) were deduced using MacVector 7. Deduced amino
acid sequences of the Myotis polyomavirus genes for Large T-antigen,
Inoculation of cultured cells. Lamb and pig kidney cells, baby Small t-antigen, VP1 and VP2,3 were compared with those of other
hamster kidney cells (BHK-21), chicken fibroblasts and quail polyomaviruses using the PHYLIP 3.63 package. The complete
fibroblasts (QT-35) were obtained from Marta Sabara (CFIA) and nucleotide sequence of the Myotis polyomavirus and deduced amino
mouse fibroblasts (NIH-3T3), African green monkey (Vero) cells and acid sequences of the deduced proteins were submitted to GenBank
bat tracheal (BT) cells were obtained from James Strong (Public and assigned the accession number FJ188392.
Health Agency of Canada, NML). All cells were grown in medium
normally used to propagate the cells.
Tissue homogenates were diluted 1/10 and 1/100 in BA and 100 ml of RESULTS
undiluted or diluted homogenate was added to cultured cells in 24-
well culture dishes. After incubating for 1 h at 37 uC, the inocula were We attempted to detect viruses in Canadian bats by using
removed and replaced with 1 ml medium containing 5 % fetal bovine two complementary strategies: to isolate virus in a variety
serum. Cells were observed for cytopathic effect (CPE) every 2 days. of cultured cells and to screen samples by PCR using
Any morphological differences or differences in the colour of the primers designed to amplify nucleic acids from members of
growth medium from mock-infected cells were considered a CPE.
selected families and genera of viruses and from individual
After 3 days of incubation, cultures inoculated with the lowest
dilution of homogenate that contained viable cells were dispersed by viruses. The target viral families and genera were selected
trypsinization and re-plated. Inoculated cells were passaged in this either because of a precedent for members spilling over
manner five times and were discarded if no obvious CPE was into humans or because the tests were available in our
observed. Samples from supernatants from cultures that showed CPE laboratory, thus allowing us to undertake a broad general-
were transferred to electron microscope grids and screened for virus ized screening. In positive control reactions, the primers
particles by the NML electron microscopy laboratory. detected nucleic acids from representative target viruses.
Extraction of nucleic acids. Total nucleic acids were extracted from
140 ml tissue homogenate using the QIAamp viral RNA mini kit Live-trapped M. lucifugus
(Qiagen) following the manufacturer’s instructions [this kit purifies
viral DNA as well as RNA (Allan Grolla, personal communication)]. We examined 31 live-trapped M. lucifugus (Table 1).
An additional washing step with buffer AW1 (Qiagen) was included. Twenty-one bats were collected from a mating swarm in
Before extraction, all samples were seeded with bacteriophage MS2 September 2007. Of these, thirteen were female and seven
RNA as an internal control. Nucleic acid samples for PCR for DNA were male (one unknown). An additional five bats were
viruses and for the two-step real-time PCR for West Nile virus were
collected from each of two cave hibernacula in May 2008. All
used directly. For PCR for RNA viruses, RNA was converted to cDNA
using the Qiagen Sensiscript RT kit with random nonamers (Gene of these bats were male. Various tissues from individual bats
Link) as primers. Control RNA from Hendra, Nipah, measles, SARS were pooled and attempts were made to isolate virus using
corona and rabies viruses were obtained from NML laboratories. cultured cells. However, we observed no virus particles by
electron microscopy, even after repeated attempts to detect
PCR. The primer sequences used for PCR and RT-PCR are provided virus in tissue culture supernatants and concentrated
in Supplementary Table S1 (available in JGV Online). PCRs for material. We did, however, detect virus infection by PCR
corona, lyssa, flavi, bunya, Cache Valley and morbilli viruses were
analysis of nucleic acids purified from tissue homogenates of
performed with ABS AmpliTaq (Applied Biosystems) according to
the manufacturer’s instructions. The amplification process included seven of the 31 live-trapped M. lucifugus examined (Table 1).
holding the samples at 94 uC for 15 min; 18 ‘touch down’ cycles of We determined the nucleotide sequence of the PCR
94 uC for 15 s, 55–37 uC (with a 1 uC decrease in temperature per products, which confirmed that four of the viruses were
cycle) and 72 uC for 30 s; 30 cycles of 94 uC for 15 s, 55 uC for 30 s polyomavirus while three were coronavirus. All four bats
and 72 uC for 30 s; 72 uC for 7 min. Secondary amplifications for from which we detected polyomavirus were females.
corona, lyssa and morbilli viruses were performed on 1 ml of the
primary PCR product using the same amplification conditions. PCR
products were analysed by electrophoresis on 2 % agarose gels. In Bats from the Centre for Expertise in Rabies and
some gels, PCR products from control viral cDNAs were included. bats collected from a wind energy facility
PCR products of the expected size were submitted to the NML
genomics core facility and sequences were analysed by BLAST to The Centre for Expertise in Rabies provided 84 bat
determine if they resembled known viral sequences. Bacteriophage carcasses representing four genera and six species

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V. Misra and others

Table 1. Virus nucleic acids detected in live-trapped M. lucifugus veterinarian. No viruses were detected in these animals
(total of 100).
NVD, No virus was detected.

Bat no. Sex* CaveD Virus detectedd Myotis polyomavirus

1 F St G NVD Polyomavirus DNA was detected using primers designed
2 F St G NVD from a conserved portion of the VP1 gene of polyoma-
3 F St G NVD viruses. Using primers designed from the nucleotide
4 U St G NVD sequence of the VP1 fragment, we amplified the remainder
5 M St G NVD
of the viral genome from three bats. The sequence of the
6 M St G NVD
entire genome was determined and putative ORFs for
7 F St G NVD
major polyomavirus proteins (VP1, VP2 and 3, Large T
8 M St G NVD
antigen and Small t-antigen) were deduced. The derived
9 M St G NVD
amino acid sequences for these proteins were compared
10 F St G Polyoma
11 F St G Polyoma
with those of all other known polyomaviruses. Only VP1
12 F St G Polyoma sequences were available for the goose polyomavirus.
13 F St G NVD Fig. 1 depicts phylogenetic trees of the polyomavirus viral
14 M St G Corona proteins based on a parsimony analysis of the amino acid
15 F St G NVD
sequences of the proteins. While there were some
16 F St G NVD
variations in the groupings when the sequences of the
17 M St G NVD
different proteins were compared, all four proteins of the
18 F St G NVD
Myotis bat virus were closely related to those of the mouse
19 F St G NVD
20 M St G NVD
pneumotropic (Kilham) and squirrel monkey viruses.
21 F St G Polyoma
22 M Dale’s NVD Myotis coronavirus
23 M Dale’s NVD
24 M Dale’s Corona We amplified a 500 bp segment of RNA from a coronavirus
25 M Dale’s NVD in three M. lucifugus bats using primers designed to amplify
26 M Dale’s NVD a portion of the coronavirus RNA polymerase gene. Bat no.
27 M Abyss NVD 14 was trapped in September 2007, while bats 24 and 30 were
28 M Abyss NVD collected in May 2008; the three bats were from different
29 M Abyss NVD caves (Table 1). The sequence of the amplified portion of the
30 M Abyss Corona coronavirus gene most closely resembled those of the
31 M Abyss NVD RMBCV sequence detected in M. occultus (Dominguez et
al., 2007) and E. fuscus in Colorado (Dominguez et al.,
*F, Female; M, male; U, unknown. 2007). A comparison of the sequences (Fig. 2) using
DBats were collected at three caves: St George (St G), Dale’s and CLUSTAL_X showed that the sequences of viruses from M.
Abyss. Bats from St George cave were trapped during a mating swarm lucifugus bats 14 and 24 were very similar. All three
in September 2007. Bats from Dale’s and Abyss caves were collected in
sequences were closely related to sequences amplified from
May 2008.
M. occultus but had several differences from those amplified
dPolyoma or corona viruses were detected by PCR in tissue lysates of
from E. fuscus (Dominguez et al., 2007).
seven of 31 animals.
When the 500 bp segment from the Myotis coronavirus was
compared with corresponding sequences from coronaviruses
detected in bats in North America, Europe and South-East Asia
(Supplementary Table S2, available in JGV Online). These (Fig. 3), the sequences of the Myotis coronavirus most closely
bats were submitted to the Centre for Rabies Testing and
resembled sequences from Group 1 coronaviruses, including
were found to be negative for rabies virus. The carcasses,
viruses from the other North American bats and Asian bats
which had been stored frozen at 220 uC, were thawed
Rhinolophus sinicus, Myotis ricketti and Miniopterus australis
overnight at room temperature before dissection. We
(Woo et al., 2006). In our analysis, the sequences of
detected polyomavirus in a pool from two female M.
coronaviruses detected in European bats (Gloza-Rausch et al.,
californicus bats submitted to the Centre from British
2008) formed a distinct clade within the group 1 coronaviruses.
Columbia. We also obtained 15 bat carcasses representing
the three migrating species (Las. cinereus, L. noctivagans
and Las. borealis) from a wind plant near Winnipeg. The
carcasses were collected in autumn 2007 and frozen at
DISCUSSION
220 uC until the tissues were analysed by PCR. An The objective of the project was to screen bats in the province
additional Las. cinerus bat was submitted by a local of Manitoba and other regions of Canada for viruses. Most of

2018 Journal of General Virology 90

 Viruses in bats of Canada

Fig. 1. Phylogenetic relationships between the amino acid sequences of VP1, VP2,3, Large T-Antigen and Small t-antigen of
polyomaviruses. The sequences for Myotis polyomavirus (MyPyV, boxed) were derived as described in Methods, the others
were obtained from GenBank (see Supplementary Table S3). The trees for each of the four proteins were generated using
SEQBOOT, Protpars and CONSENSE. The consensus trees were based on a maximum-parsimony analysis using 1000 bootstrap
replicates of the data. For each tree, the bootstrap value (of 1000 bootstrap replicates) for a specific branch is indicated either
above or below the branch. Note that the values are given as a percentage. Values ,70 % are generally not given except where
the value is of some particular interest. AGMonkey, African green monkey polyomavirus; JC, JC virus; SV40/12, Simian virus 40/
12; WU, WU polyomavirus; KI, KI polyomavirus.

the viruses were detected in live-trapped M. lucifugus. Seven for several months and it appeared that considerable time had
of the 31 animals (over 22 %) examined yielded virus. With elapsed between the death of these animals, many by trauma,
the exception of two M. californicus, no viruses were detected and freezing. Bats from the wind plant may have been
in any of the bats obtained from the rabies laboratory or the exposed to the elements for as long as 24 h before being found
wind plant. These 100 bats represented eleven species. Our by searchers and collected. In contrast, the live-trapped bats
failure to detect viruses in these animals likely reflects the were sampled within a day of euthanasia and maintained on
condition of the animals. All were thawed after being frozen ice during that period.

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V. Misra and others

Fig. 2. Relationship of sequences from a conserved segment of

the RNA polymerase gene of RMBCV detected in North American
bats. The nucleotide sequences for virus detected in M. lucifugus
Fig. 3. Phylogenetic relationship between the nucleotide
(M. luc, boxed; these are MyPyV sequences) were derived as
sequences of an approximately 250 bp segment of the RNA
described in Methods, the others [M. occultus (M. occ) and E.
polymerase gene from several bat coronaviruses. The nucleotide
fuscus (E. fus)] were obtained from GenBank (see Supplementary
sequences for virus detected in M. lucifugus (MyPyV strains;
Table S3). GenBank accession numbers are given in parentheses.
boxed) were derived as described in Methods, others were
Only the corresponding sequences present in all samples were
obtained from GenBank (see Supplementary Table S3).
included in the CLUSTAL_X alignment. The unrooted trees were
Sequences corresponding to those amplified by PCR were
drawn in NJPlot using data from CLUSTAL_X.
extracted from some sequences before alignment. Sequences of
bat coronaviruses detected in North America (Roman type), Asia
(italics) and Europe (underlined) are represented. The bat species
Using a strategy that exploited the circular nature of in which the viruses were detected and their accession numbers
polyomavirus genomes (Johne et al., 2006), we recovered are: Pabr, Pipistrelles abramus, DQ249216; Tpac, Tylonycteris
the entire genome of the virus detected in two Myotis pachypus, DQ075642; Mdau, M. daubentonii, D8.38, EU375874;
species. Analysis of the complete sequence of the viral D8.32, EU375875; Mdas, M. dasycneme, D3.33, EU375854;
genomes recovered from three animals revealed an as-yet D5.17, EU375861; Pnat, P. nathusii, D5.16, EU375864; D5.73,
EU375869; Mbec, M. bechsteinii, D6.6, EU375865; Ppyg, P.
undescribed polyomavirus. We suggest that the virus
pygmaeus, D5.71, EU375868; D5.85, EU375870; Mric, M.
should be called Myotis polyomavirus (MyPyV). The
ricketti, DQ249224; Maus, M. australis, DQ249228; Efus,
genome of MyPyV appeared to have the same arrangement
Eptesicus fuscus, EF544566; Mocc, M. occultus, 3, EF544567;
of genes as other polyomaviruses, with potential ORFs for 11, EF544563; 48, EF544565. The unrooted trees were drawn in
the capsid proteins VP1, VP2 and VP3. ORFs with NJPlot using data from CLUSTAL_X.
potential splice sites, the non-structural proteins Small t
and Large T antigens were also present. The genome
contained other small ORFs, including one for a potential
‘agno’ protein. However, we could not detect significant pups. In contrast with the mammalian viruses, at least two
similarity between the deduced amino acid sequence of of the avian viruses cause acute, frequently fatal inflam-
these ORFs and peptides identified for the better- matory disease in some species of psittacine birds and geese
characterized polyomaviruses. (Johne & Müller, 2007). The cycle of infection of most
polyomaviruses in natural hosts has not been determined.
Some mammalian polyomaviruses cause tumours when However, the avian polyomavirus appears to have a unique
susceptible rodents are infected experimentally and the form of reverse vertical transmission in the European pied
newly discovered Merkel polyomavirus is associated with flycatcher, where blowfly larvae transmit the virus to
the aggressive Merkel cell skin cancer in humans (Feng
nestlings, which then infect the parents through the faecal–
et al., 2008). In addition, both the mouse polyomavirus
oral route (Potti et al., 2007).
and pneumotropic virus can cause disease when inoculated
into newborn mice. However, the mammalian polyoma- Like other mammalian polyomaviruses in natural hosts,
viruses are not generally associated with acute disease in MyPyV appears not to cause disease in adult M. lucifugus.
natural non-immunocompromised hosts. While the nat- We detected the virus in apparently healthy bats in a
urally occurring mouse virus does retain its ability to cause mating swarm. While the number of bats we examined was
fatal disease in newborn mice, infection under natural too low to reach a definitive conclusion, it is interesting
conditions in feral mice does not result in disease (Carroll that we only detected the virus in females of two Myotis
et al., 2007), possibly because of the simultaneous species. Myotis females rear pups in maternity roosts with
transmission of virus and maternal immunity to newborn little input into offspring care by males. It is tempting to

2020 Journal of General Virology 90

 Viruses in bats of Canada

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