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Address for correspondence: Fang Huang, Beijing Center for Disease
Prevention and Control, Institute for Communicable Disease Control
and Prevention, No. 16, Hepingli Middle Av, Dongcheng District,
Beijing 100013, China; email: firstname.lastname@example.org
Recent Findings of Potentially
Lethal Salamander Fungus
David Lastra González, Vojtech Baláž,
Milič Solský, Barbora Thumsová,
Krzysztof Kolenda, Anna Najbar,
Bartłomiej Najbar, Matej Kautman, Petr Chajma,
Monika Balogová, Jiří Vojar
Author aliations: Czech University of Life Sciences, Prague,
Czech Republic (D. Lastra González, M. Solský, B. Thumsová,
P. Chajma, J. Vojar); University of Veterinary and Pharmaceutical
Sciences, Brno, Czech Republic (V. Baláž, M. Kautman);
University of Wrocławski, Wroclaw, Poland (K. Kolenda,
A. Najbar); University of Zielona Góra, Lubuskie, Poland
(B. Najbar); Slovak Academy of Sciences, Košice, Slovakia
(M. Kautman); Pavol Jozef Šafárik University in Košice, Košice
The distribution of the chytrid fungus Batrachochytrium
salamandrivorans continues to expand in Europe. During
2014–2018, we collected 1,135 samples from salamanders
and newts in 6 countries in Europe. We identied 5 cases of
B. salamandrivorans in a wild population in Spain but none
in central Europe or the Balkan Peninsula.
Chytridiomycosis, an amphibian disease caused by the
chytrid fungi Batrachochytrium dendrobatidis and B.
salamandrivorans, is responsible for declines of amphib-
ian populations worldwide (1). The recently discovered B.
salamandrivorans (2) is severely impacting salamanders
and newts in Europe (3,4). This emerging fungal pathogen
infects the skin of caudates and causes lethal lesions (2). It
most likely was introduced to Europe by the pet salamander
trade from Southeast Asia (3). In Europe, the Netherlands,
Belgium, and Germany have conrmed B. salamandriv-
orans in wild caudates; the United Kingdom, Germany,
and Spain have conrmed the fungus in captive animals
(5,6). Several countries have established trade regulations
(5) and a recent European Union decision, no. 2018/320,
implements measures to protect against the spread of B.
salamandrivorans via traded salamanders (7). The World
Organisation for Animal Health listed infection with B.
salamandrivorans as a notiable disease in 2017. In ad-
dition to controlling the amphibian pet trade, surveillance
of the pathogen is urgently needed to establish disease in-
tervention strategies in aected areas and prevention in B.
During 2014–2018, we collected 1,135 samples directly
for the detection of B. salamandrivorans or as a part of unre-
lated studies. Samples came from 10 amphibian species at 47
sites in 6 countries in Europe. Most samples came from the
re salamander, Salamandra salamandra, which is a known
suitable host for B. salamandrivorans (3), and the palmate
newt, Lissotriton helveticus, which is known to be resistant
to B. salamandrivorans (Appendix Table 1, http://wwwnc.
Most samples were skin swabs collected by following
the standard procedure for sampling of amphibian chytrid
fungi (8). A smaller portion of samples was toe clippings
(Appendix Table 2). We extracted genomic DNA following
the protocol of Blooi et al. (9), and 2 laboratories with dif-
ferent equipment tested for B. salamandrivorans. Samples
from Spain and the Czech Republic initially were analyzed
at the Czech University of Life Sciences (Prague, Czech
1416 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 7, July 2019
Republic) by standard PCR with B. salamandrivorans–
specic primers STerF and STerR, as described by Martel
et al. (2), with subsequent electrophoresis on the ampli-
ed target. We reanalyzed samples that produced positive
or equivocal results by using duplex quantitative PCR
(qPCR) for B. dendrobatidis and B. salamandrivorans (9)
at the University of Veterinary and Pharmaceutical Scienc-
es (Brno, Czech Republic). Trenton Garner of the Institute
of Zoology, Zoological Society of London (London, Eng-
land), provided DNA for quantication standards of the B.
dendrobatidis GPL lineage, strain IA042, and An Martel
of Ghent University (Ghent, Belgium) provided quantica-
tion standards of B. salamandrivorans.
We directly analyzed samples from other countries by
qPCR. We used negative and positive controls for standard
PCR analyses and quantication standards for qPCR analy-
ses. For B. dendrobatidis– or B. salamandrivorans–posi-
tive sites, we estimated prevalence and Bayesian 95% CIs
using 3 parallel Markov chains with 2,000 iterations each,
a burn-in of 1,000 iterations, and no thinning (Appendix
Table 1). We performed all statistical analyses in R 3.3.1
using the R2WinBUGS package and WinBUGS 1.4.3 (10).
Samples from 5 L. helveticus newts tested positive
for B. salamandrivorans, implying that this species is
not resistant to this fungus as previously indicated by ex-
perimental exposures (3). The positive cases were found
in populations from an isolated area encompassing 2 dif-
ferent regions in northern Spain, Cantabria and Asturias,
with remote human populations. Four cases were found in
livestock drinking troughs located 150–1,000 m above sea
level, and 1 case was found in a pond in a private garden,
30 km from the nearest recorded case. We did not nd B.
salamandrivorans–positive cases in consecutive locations
during our monitoring.
Although B. salamandrivorans cases have been re-
ported in captive salamanders (6), our reported cases were
>1,000 km from any area of known B. salamandrivorans
occurrence (7). We also detected B. dendrobatidis by du-
plex qPCR in 11 samples from 3 newt species (L. helve-
ticus, L. vulgaris, and Triturus cristatus) from Spain and
Montenegro and 1 captive Cynops ensicauda newt from the
Czech Republic. The B. dendrobatidis–positive cases did
not involve co-infection with B. salamandrivorans.
We conrmed that the known distribution of B. sala-
mandrivorans continues to expand in Europe, indicating
that this fungus might be capable of dispersing over long
distances (4), might be introduced by humans, or might
even have been circulating in this geographic range with
no detected deaths. Our results should alert the research
and conservation community and motivate urgent action to
identify regions with early emergence of the disease and
implement mitigation measures to prevent further spread
of this deadly pathogen.
We thank the Cantabria delegation of SEO/Birdlife; Fundación
Zoo Santillana del Mar; workers from Marismas de Santoña,
Victoria y Joyel Natural Park, with special thanks to
Carlos Rubio; Pepo Nieto, Pedro Barreda, and his family;
Elena Kulikova and Wiesław Babik; and also our friends
Daniel Koleška, Kamila Šimůnková, Tomáš Holer, and
Daniela Budská for eldwork.
This work was performed with permission from the Nature
Conservation Agency of the Czech Republic; Agency for Nature
and Environment Protection of Montenegro permit no. 02 Broj
UPI–321/4; Ministry of Environment of the Slovak Republic,
permit no. 4924/2017–6.3; the Endangered Species Section of
Environmental Service of Cantabria, Spain, permit no. EST–
419/2017–SEP; the Environmental Service of Castilla y León,
Spain, permit no. EP/LE/233/2017; Department of Nature
Conservation of Poland, permit nos. DZP-WG.6401.02.7.2014.
JRO, WPN.6401.211.2015.MR.2, 78/2014, and 68/2015; the
Ministry of Protection of Environment of Croatia, permit no.
UP/I–612–07/169–48/68; and agreements from other agencies,
including Red Cambera, special thanks to Sergio Tejón and Tomás
González; Fondo para la Protección de los Animales Salvajes; and
Fundación Naturaleza y Hombre, Spain. The study was supported
by the Czech University of Life Sciences, Prague, Czech Republic
(grant nos. 20174218 and 20184247) and the Internal Grant
Agency of the University of Veterinary and Pharmaceutical
Sciences, Brno, Czech Republic (grant no. 224/2016/FVHE).
K.K. was supported by MNiSW grant for Young Scientists no.
0420/1408/16; A.N. was supported by grant no. DS 1076/S/
IBŚ/2014 and MNiSW grant for Young Scientists no. 0420/1409/16.
About the Author
Mr. Lastra González is a PhD candidate at Czech University
of Life Sciences, Prague. His research focuses on amphibian
conservation and emerging infectious diseases that aect them.
1. Berger L, Roberts AA, Voyles J, Longcore JE, Murray KA,
Skerratt LF. History and recent progress on chytridiomycosis in
amphibians. Fungal Ecol. 2016;19:89–99. http://dx.doi.org/
2. Martel A, Spitzen-van der Sluijs A, Blooi M, Bert W,
Ducatelle R, Fisher MC, et al. Batrachochytrium salamandrivorans
sp. nov. causes lethal chytridiomycosis in amphibians. Proc Natl
Acad Sci U S A. 2013;110:15325–9. http://dx.doi.org/10.1073/
3. Martel A, Blooi M, Adriaensen C, Van Rooij P, Beukema W,
Fisher MC, et al. Recent introduction of a chytrid fungus endangers
Western Palearctic salamanders. Science. 2014;346:630–1.
4. Stegen G, Pasmans F, Schmidt BR, Rouaer LO, Van Praet S,
Schaub M, et al. Drivers of salamander extirpation mediated by
Batrachochytrium salamandrivorans. Nature. 2017;544:353–6.
Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 7, July 2019 1417
5. European Food Safety Authority Panel on Animal Health and
Welfare, More S, Angel Miranda M, Bicout D, Bøtner A,
Butterworth A, et al. Risk of survival, establishment and spread of
Batrachochytrium salamandrivorans (Bsal) in the EU. EFSA
Journal. 2018;16:5259. http://dx.doi.org/10.2903/j.efsa.2018.5259
6. Fitzpatrick LD, Pasmans F, Martel A, Cunningham AA.
Epidemiological tracing of Batrachochytrium salamandrivorans
identies widespread infection and associated mortalities in private
amphibian collections. Sci Rep. 2018;8:13845. http://dx.doi.org/
7. Commission Implementing Decision (EU) 2018/320 of 28
February 2018 on certain animal health protection measures for
intra-Union trade in salamanders and the introduction into the
Union of such animals in relation to the fungus Batrachochytrium
salamandrivorans. Ocial Journal of the European Union.
2018;L62:18–33 [cited 2018 Jun 1]. http://data.europa.eu/eli/
8. Hyatt AD, Boyle DG, Olsen V, Boyle DB, Berger L, Obendorf D,
et al. Diagnostic assays and sampling protocols for the detection of
Batrachochytrium dendrobatidis. Dis Aquat Organ. 2007;73:175–
9. Blooi M, Pasmans F, Longcore JE, Spitzen-van der Sluijs A,
Vercammen F, Martel A. Duplex real-time PCR for rapid
simultaneous detection of Batrachochytrium dendrobatidis and
Batrachochytrium salamandrivorans in amphibian samples. J Clin
Microbiol. 2013;51:4173–7. http://dx.doi.org/10.1128/JCM.02313-13
10. Lunn DJ, Thomas A, Best N, Spiegelhalter D. WinBUGS–
a Bayesian modelling framework: concepts, structure, and
extensibility. Stat Comput. 2000;10:325–37. http://dx.doi.org/
Address for correspondence: David Lastra González, Faculty of
Environmental Sciences, Czech University of Life Sciences,
Kamýcká 129, 165 21 Prague–Suchdol, Czech Republic;
Fever Virus Genome in Tick
from Migratory Bird, Italy
Elisa Mancuso, Luciano Toma, Andrea Polci,
Silvio G. d’Alessio, Marco Di Luca,
Massimiliano Orsini, Marco Di Domenico,
Maurilia Marcacci, Giuseppe Mancini,
Fernando Spina, Maria Goredo,
Author aliations: Istituto Zooprolattico Sperimentale
dell’Abruzzo e del Molise “G. Caporale,” Teramo, Italy
(E. Mancuso, A. Polci, S.G. d’Alessio, M. Orsini,
M. Di Domenico, M. Marcacci, G. Mancini, M. Goredo,
F. Monaco); Istituto Superiore di Sanità, Rome, Italy (L. Toma,
M. Di Luca); Istituto Superiore per la Protezione e la Ricerca
Ambientale, Bologna, Italy (F. Spina)
We detected Crimean-Congo hemorrhagic fever virus in a
Hyalomma rupes nymph collected from a whinchat (Saxi-
cola rubetra) on the island of Ventotene in April 2017. Partial
genome sequences suggest the virus originated in Africa.
Detection of the genome of this virus in Italy conrms its
potential dispersion through migratory birds.
Crimean-Congo hemorrhagic fever virus (CCHFV) is a
vectorborne virus responsible for severe illness in hu-
mans, whereas other mammals usually act as asymptomatic
reservoirs. The virus is transmitted through tick bites or by
direct contact with blood or body uids of infected verte-
brate hosts. CCHFV, an Orthonairovirus within the Nairo-
viridae family, has a negative-sense tripartite RNA genome
characterized by high genetic diversity. The sequences of
the circulating strains cluster in 6 genotypes (I–VI) reect-
ing their geographic origin; worldwide distribution is the
result of ecient dispersion through migratory birds, hu-
man travelers, and the trade and movement of livestock and
wildlife (1,2). In Europe, CCHFV distribution was limited
to the Balkan region until 2010, when the virus was iden-
tied in ticks collected from a red deer (Cervus elaphus)
and, 6 years later, in 2 autochthonous human cases in the
same region of Spain (3). Sequences from the Iberia strains
clustered in the Africa genotype III (4), supporting the hy-
pothesis of CCHFV dispersion through ticks hosted by mi-
The role of birds in the potential spread of the virus
was conrmed by CCHFV detection in ticks collected from
migratory birds in Greece in 2009 (5) and Morocco in 2011
(6). Because Italy hosts an intense passage of birds migrat-
ing along major routes connecting winter quarters in Africa
and breeding areas in Europe, the country is potentially
exposed to the risk for virus introduction. We report the
detection of CCHFV RNA in a tick collected in Italy from
a migratory bird.
We conducted tick sampling during March–May 2017
on the island of Ventotene, where a ringing station has been
operating since 1988 as part of the Small Islands Project, a
large-scale and long-term eort to monitor spring migra-
tions of birds across the central and western Mediterranean.
We ringed 5,095 birds and checked ≈80% for ectoparasites.
We collected 14 adults, 330 nymphs, and 276 larvae from
268 passerines belonging to 28 species; 18 species were
trans-Saharan migrants. We stored ticks in 70% ethanol
until morphologic identication and assignment to a genus
or, whenever possible, a species (7). We then individually
1418 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 25, No. 7, July 2019