Detection of bovine torovirus in fecal specimens from calves
with diarrhea in Turkey
Received: 8 October 2013 / Accepted: 31 December 2013 / Published online: 14 January 2014
Ó Springer-Verlag Wien 2014
Abstract Bovine torovirus (BToV), a member of the
family Coronaviridae, is an established gastrointestinal
infectious agent in cattle. In this study, we performed a survey
to detect BToV in Turkey between 2009 and 2011 using 235
fecal samples from neonatal calves with diarrhea that were
analyzed by the nested reverse transcription (RT) PCR
method using primers located in the consensus sequences of
the BToV membrane (M) gene. The BToV M gene was
detected in 4.7 % (11/235) of the samples using the nested
RT-PCR method. The nucleotide sequences of partial M
fragments from the BToV isolates, including the newly
identiﬁed Turkish isolates, showed more than 96 % identity.
The result indicates that BToV is one of the pathogens that
contribute to neonatal calf diarrhea cases in Turkey.
Torovirus is a genus of the family Coronaviridae, order
Nidovirales, and the members of this genus are enveloped,
single-stranded, positive-sense RNA viruses [14, 28, 31].
Toroviruses cause gastroenteritis in mammals and have
been detected in humans, horses, cattle, and swine
worldwide [1–3, 6, 9, 13, 16]. BToV was ﬁrst detected in
the USA during an outbreak of diarrhea in cattle in 1979
. Epidemiological studies of BToV have shown that it
is widespread throughout the world [7, 10–13, 15, 17, 22–
26, 30, 32, 34]. However, there have not been any reports
of BToV infections in Turkey.
Human and bovine toroviruses are difﬁcult to grow in
cell culture. However there has been one report on a BToV
(Aichi/2004) that was isolated in a human rectal adeno-
carcinoma cell line (HRT-18) from the ileum content of a
calf with diarrhea . Since growth of BToV in cell
culture was established just a few years ago, there are only
a limited number of sequences of BToV available in the
GenBank database [4, 5, 8–13, 15, 25, 27].
The aim of this study was to determine the presence of
bovine toroviruses in diarrheic feces from calves in Turkey.
This report also describes the ﬁrst detection of BToV in
Turkey, using the RT-PCR technique, along with the
genetic diversity of the BToV strains based on partial
BToV membrane (M) gene sequences.
Materials and methods
A total of 235 diarrheic fecal specimens were collected from
calves with diarrhea from 28 provinces of Turkey between
2009 and 2011. Figure 1
indicates the locations where samples
were taken and where they were found to be positive for BToV.
The fecal samples were diluted in a 109 volume of 0.01 M
phosphate-buffered saline (0.138 M NaCl, 0.0027 M KCl,
The Pendik Veterinary Control and Research Institute,
H. Is¸ıdan (&)
Department of Virology, Cumhuriyet University Faculty of
Veterinary Medicine, 58140 Sivas, Turkey
Department of Virology, Erciyes University Faculty of
Veterinary Medicine, 38039 Kayseri, Turkey
Arch Virol (2014) 159:1623–1627