ISSN 10674136, Russian Journal of Ecology, 2015, Vol. 46, No. 5, pp. 490–493. © Pleiades Publishing, Ltd., 2015.
Original Russian Text © K.A. Kravchenko, A.S. Vlashchenko, O.V. Prilutskii, A.S. Prilutskaya, 2015, published in Ekologiya, 2015, No. 5, pp. 397–400.
In 2006, a pandemic of previously unknown fungal
disease suddenly broke out in caves of North America
(Cohn, 2008; Blehert et al., 2009), affecting bats dur
ing winter hibernation (Gargas et al., 2009; Cryan
et al., 2010). Between 2006 and 2012, six million bats
of ten species died (http://www.batcon.org/index.
and the infection was spreading from the east coast of
the United States into the continent for several hun
dred kilometers per year (Turner, 2011; Raloff, 2011).
The mass mortality of bats from this disease and the
high rate of its spread attracted close attention from
researchers. In 2009, the causative agent of this dis
ease, named White Nose Syndrome (WNS), was iden
Blehert et Gargas (Gar
gas et al., 2009), a fungal species previously unknown
to science whose mycelium grows into the surface
body tissues of hibernating bats (Carol et al., 2009).
The main hypothesis concerning the origin of this
pandemic outbreak—that the fungus was introduced
from Europe (Wibbelt et al., 2010; Puechmaille et al.,
2010)—was confirmed in subsequent morphological,
immunological, and genetic studies (Wernecke et al.,
2012). In 2008 to 2011, largescale surveys of under
ground bat winter roosts were conducted in Western
and Central Europe in order to find bats affected by
WNS (Wibbelt et al., 2010; Puechmaille et al., 2011).
The easternmost record of
was made in
Gumenetskie Shtol’ni, a system of adits in Khmelnitsky
oblast, Ukraine (Puechmaille et al., 2011). Information
on the occurrence of this fungus in eastern and north
ern regions of Europe was absent.
The purposes of our study were as follows: (1) to
evaluate the occurrence of
ground cavities of Ukraine and European Russia,
(2) to compare the efficiency of its detection by tradi
tional morphological and modern genetic methods,
and (3) to present preliminary data on the mycobiota
of surveyed caves.
Samples for analysis were taken in 2010 and 2010
from seven manmade caves serve as major winter
roosts for bats and are located in four regions of
Ukraine and European Russia (Table 1). Special cen
suses of hibernating bats were taken only if compre
hensive published data were not available. Spores were
trapped on the surface of glass slides (below, referred to
as spore traps) or Petri dishes coated with agar medium
(Flannigan, 1997). In both cases, Sabouraud pep
tone–glucose agar (SAB) and cornmeal agar (CMA)
was used (Bills and Foster, 2004). Spore traps were
fixed on cave walls and exposed for 60 days, while Petri
dishes were exposed for 1–3 h. Both sampling meth
ods were tested in Liptsy1 and Liptsy2 caves in June
to August 2010; samples from other caves were taken
only with Petri dishes.
In Liptsy1 and Liptsy2, ten Petri dishes and ten
spore traps were exposed in each cave, half of them
with CMA and the other half with SAB. Subsequent
samples were taken only with CMA, which proved to
be more effective. Twenty dishes were exposed in caves
of Leningrad oblast (12 in Levoberezhnaya and 8 in
Plyazhnaya); 15 dishes, in Zaporozhye oblast (12 in
Skelyanskie Mines and 3 in Mayachanskaya); and
16 dishes, in Gumenetskie Mines, where samples
(scrapings) were also taken directly from the skin of
affected bats, from patches with white fungal over
After sampling, Petri dishes were incubated in a
, and examined for fungal growth
every 4–5 days, recording characteristics of colonies.
Sporebearing colonies were subcultured to new
dishes with CMA to obtain pure colonies. In some
cases two to three passages were made using stab inoc
ulation (Bilai, 1982).
Skin scrapings were plated in Petri dishes with
CMA and incubated in a cooler. Pure cultures were
examined to record their color, texture, features of
mycelium growth, and spore production. All samples
were identified to the level of genus (in some cases, of
A Search for
, a Dangerous Pathogen of Bats,
in Caves of Eastern Europe
K. A. Kravchenko
, A. S. Vlashchenko
, O. V. Prilutskii
, and A. S. Prilutskaya
Feldman Ecopark Bat Rehabilitation Center, Kievskoe sh. 12, Lesnoe, Kharkov oblast, 62340 Ukraine
Karazin Kharkov National University, pl. Svobody 4, Kharkov, 61022 Ukraine
Received March 5, 2014
, fungi, caves, bats, Ukraine, Russia