Animal Conservation.

Print ISSN 1367-9430

SUPPLEMENT ARTICLE

Widespread occurrence of the amphibian chytrid fungus

in Kenya
J. Kielgast1, D. Rödder2, M. Veith2 & S. Lötters2
1 Department of Biology, University of Copenhagen, Copenhagen, Denmark
2 Department of Biogeography, Trier University, Trier, Germany

Keywords
amphibian decline; Batrachochytrium
dendrobatidis; chytridiomycosis; ‘out of
Africa’; endemic; enzootic; Kenya; Africa.
Correspondence
J. Kielgast, Department of Biology, Section
for Evolution and Microbiology, University
of Copenhagen, Universitetsparken 15,
2100 Copenhagen, Denmark.
Email: jkielgast@snm.ku.dk
Received 15 February 2009; accepted 15
June 2009
doi:10.1111/j.1469-1795.2009.00297.x
Abstract
Amphibians at the global scale are dramatically declining and the pathogenic
fungus Batrachochytrium dendrobatidis (Bd) has been suggested to be an important
driver in this biodiversity crisis. Increasing evidence points towards the global
emergence of Bd being a panzootic caused by pathogen pollution. Africa has been
suggested to be the origin of the pathogen but remains one of the least-studied
areas. We have conducted the most comprehensive survey on the continent to date
focusing on Kenya for investigating taxonomic and environmental components in
the distribution of Bd in tropical Africa. Eleven sites along a 770 km transect from
the coast up to the border of Uganda were surveyed. Using quantitative PCR, we
screened 861 samples from 23 different species in nine genera. The pathogen was
confirmed at all studied sites, with an overall prevalence of 31.5%. No dead or
symptomatic specimens were found and no declines have been reported in the
region so far. Both prevalence and parasite load ranged from the detection limit to
some of the highest ever reported. The parasite load showed a significant
taxonomic bias and a strong inverse correlation with temperature. Our findings
suggest that Bd may be enzootic in the region. We recommend that further
research should focus on comparative experimental studies of susceptibility to Bd
in African species. Moreover, we stress the need for improved knowledge on the
conservation status of the tropical African amphibian fauna to confirm the
enzootic nature of widespread Bd infections.

spread (James et al., 2009). This is supported by empirical

Introduction
Amphibians are the most drastically declining vertebrates
on our planet, with nearly one-third of the c. 6400 known
species being threatened with extinction under the IUCN
Red List (Stuart et al., 2008). This biodiversity crisis is
developing globally at a rate only paralleled by mass extinc-
tion events in geological time (Mendelson III et al., 2006;
Wake & Vredenburg, 2008). Epizootics of the disease
chytridiomycosis, caused by the parasitic fungus Batracho-
chytrium dendrobatidis (Bd), have been proposed to play an
important role as the proximate cause of the rapid decline of
more than 200 amphibian species (Skerratt et al., 2007). This
is supported by well-documented aetiology in focal studies
of population decline in North America (e.g. Rachowicz
et al., 2006), Central America (e.g. Lips et al., 2006), Europe
(e.g. Bosch, Martı́nez-Solano & Garcı́a-Parı́s, 2001) and
Australia (e.g. Schloegel et al., 2006). Bd shows a remark-
ably low host specificity and has now been detected in more
than 400 different anuran and salamander species on all
continents on which amphibians occur (Fisher, 2008). How-
ever, molecular studies strongly indicate that Bd’s current
geographic distribution is the result of a recent panzootic
evidence of a wave-like emergence of disease in the Neo-
tropics (Lips et al., 2008).
The large-scale dissemination of Bd has been suggested to
be human mediated and data on the international trade of
amphibian species underline the potential for pathogen
pollution (e.g. Daszak et al., 2006; Fisher & Garner, 2007;
Walker et al., 2008). The point of origin is therefore a central
question towards an understanding of the epidemiology of
this disease. It has been advocated, based on evidence from
clawed frogs (genus Xenopus), that an ‘out of Africa’
scenario is plausible (Weldon et al., 2004). This was founded
on the oldest known record of Bd, a temporally constant Bd
prevalence in museum collections and disease resistance.
Moreover, a dissemination pathway exists via worldwide
trade in these frogs beginning as early as the 1930s – first
used for a pregnancy assay and subsequently as a model
animal in biological and medical teaching and research
(Weldon et al., 2004; Weldon, De Villiers & Du Preez, 2007).
The ‘out of Africa’ hypothesis has been frequently re-
ferred to in the literature. However, research on Bd has so
far suffered from a geographic bias, whereby Africa has
largely been left unstudied. Owing to the lack of apparent

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Amphibian chytrid fungus in Kenya J. Kielgast et al.

barriers to Bd within the continent, we hypothesize that an
‘out of Africa’ scenario would render widespread occurrence
of Bd in suitable habitats. Furthermore, if the host–patho-
gen system has co-evolved in the African herpetofauna,
some intrinsic factors characterizing its interactions are
likely to differ from the epizootic range, that is parameters
such as virulence and host specificity. Here we examine the
host–pathogen system in tropical Africa using landscape-
diverse Kenya as a representative. For the first time, we
investigate multiple localities and host taxa with quantita-
tive methods and present the most comprehensive survey of
Bd conducted in Africa so far.
Materials and methods
From 16 September to 22 October 2006, fieldwork was
carried out at 11 localities along a 770 km transect from the
Kenyan coast up to the border of Uganda covering semi-
humid and humid habitats at an altitudinal range of
159–3100 m a.s.l. (Fig. 1, Appendix S1). The time of sam-
pling targeted the beginning of the ‘short rains’ of the
bimodal precipitation pattern. No information on the sea-
sonality of Bd infection dynamics in Africa is available but
sampling took place corresponding well with the recommen-
dations given for sampling in Australia, for example, focus-
ing on the cold season (Kriger & Hero, 2006; Skerratt et al.,
2008). Approximately 1500 specimens were examined in situ
during the survey. A minimum of 30 individual diagnostic
samples for Bd per species and locality was targeted to
enable 95% probability of detecting a positive assuming an
underlying prevalence of 10% (following Cannon & Roe,
1982). Only post-metamorphic and adult anurans were
sampled (as caecilians were not found and salamanders are
absent). Recommended disinfection and containment pro-
cedures were followed to avoid transmission and dissemina-
tion of the pathogen (see Speare et al., 2004). Specimens
were caught by hand or, in a few cases, by dip-nets and
placed individually into plastic bags. Transmission of Bd
between individuals was eliminated by collecting the speci-
mens wearing a latex glove or a plastic bag on the hand.
Diagnostic sampling was carried out using a diagnostic fine-
tip dry swab (Medical Wire & Equipment, MW-100) by
comprehensively swabbing each specimen’s dorsum, ven-
trum, both lateral sides, the dorsal and ventral surface of
hind limbs, toes and toe webbing. A fresh pair of latex gloves
was used for every specimen to avoid contamination of
samples. Voucher specimens to validate the taxonomic
identification (following Channing & Howell, 2006) of all
sampled species (Appendix S2) were deposited at National
Museums of Kenya (NMK), Nairobi and Zoologisches
Forschungsmuseum Alexander Koenig (ZFMK), Bonn.
Swabs were stored as cool as possible in the field and
thereafter at 20 1C until processing.
For Bd analysis, 861 samples from 23 different species in
nine genera (Appendix S2) were analysed by quantitative
PCR, following the protocol of Boyle et al. (2004), but
running each sample in two replicates. Samples were identi-
fied as positive for Bd if a clear log-linear amplification was
observed for both replicates and genomic equivalents (GE)
quantified according to standards yielded above 0.5. If

Figure 1 Study sites in Kenya sampled for Batrachochytrium dendrobatidis: 1, Saiwa Swamp National Park; 2, Mt. Elgon National Park; 3,
Kakamega Forest National Reserve; 4, Thompson Falls; 5, Aberdares National Park (moorlands); 6, Aberdares National Park (Salient); 7, Tigoni
Dam; 8, Nairobi (Karens); 9, Taita Hills (Mwundanyi); 10, Taita Hills (Mwatate); 11, Shimba Hills National Parc (see Appendix S1).

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J. Kielgast et al.
between-replicate standard deviation on the GE was higher
than either of the quantified replicates, or only one replicate
amplified above the detection threshold, the sample was re-
run. If this also yielded equivocal results, the sample was
identified as negative. The mean GE of positive samples was
regarded as an index for parasite load.
Fisher’s exact test was used for comparing Bd prevalence
in the sampled genera. The parasite loads were compared by
a rank-based generalized linear model (using SAS 9.1
Statistical package). Detected GE loads within a locality
were percentile ranked by partitioning them into 100 groups
in which the smallest received a value of 0 and the largest
value received a value of 99. Thus, the influence of sample
size and locality-specific parasite abundance was removed,
enabling comparison of interspecific parasite load across
localities in a generalized linear model. The GE loads
present in the investigated genera were further compared
by the distribution-free mood’s median test using Quantita-
tive Parasitology 3.1 (http://www.zoologia.hu/qp/qp.html).
In order to assess the relationships between climate
conditions during the surveys and Bd prevalence among
sites, we extracted information on current climate (mini-
mum and maximum temperature and precipitation) and
topography from the Worldclim database, version 1.4
(http://www.worldclim.org). This is a climate model based
on the weather conditions recorded between 1950 and 2000
with a grid cell resolution of 30 arc sec (Hijmans et al., 2005)
and was created by interpolation using a thin-plate smooth-
ing spline of observed climate at weather stations with
latitude, longitude and elevation as independent variables
(Hutchinson, 1995; 2004). The relationships between local-
ity-level prevalence and environmental factors were assessed
with simple linear regressions calculated with Xlstat 2009
(http://www.addinsoft.com).
Results
All 11 sampled localities were Bd positive, with a prevalence
from 4 to 71% and an overall mean of 31.5% (Table 1,
Appendix S2). Parasite load ranged from the detection
threshold up to more than one million GE. The highest
parasite load coincided with the highest genus-specific pre-
valence at a stream habitat in the moorlands of Aberdares
National Park (Fig. 1), where only a single species, Amietia
wittei (Ranidae), was sampled. The distribution of Bd
parasite load was heavily aggregated in a small subset of
samples with extremely high GE loads as illustrated by the
variance/mean ratio (Table 1). However, a high parasite
load was generally unusual throughout the survey, as
indicated by the median mirroring central tendency for GE
load in the infected fraction of samples (Table 1). Again, the
sampled A. wittei from Aberdares National Park displayed
unusually high values with a median in the parasitized
fraction of 3966 GE. Evaluating potential taxonomic bias
in the distribution of Bd, we found a significant difference in
the proportion of infected individuals across all sampled
genera (exact two-sided, Po0.001). The parasite load simi-
larly showed a significant effect of genus (n = 861, d.f. = 8,
Animal Conservation 13, Suppl.1 (2010) 1–8
c 2009 The Authors. Journal compilation
Amphibian chytrid fungus in Kenya
w2 = 67.57, Po0.001). Hence, we can conclude that our
results strongly indicate a taxonomical component to the
abundance and intensity of infection at the time of sampling.
There was no significant difference in the median parasite
load across all genera containing more than a single infected
individual (exact two-sided, P= 0.077). Regression analysis
on climate variables indicates a negative correlation between
temperature and prevalence during the sampling period.
This pattern strongly co-varies with altitudinal distribution
and shows an increase in the frequency of Bd infection with
altitude. At the same time, the general precipitation pattern
was not found to correlate with prevalence (Fig. 2).
Discussion
Using landscape-diverse Kenya as a representative, we have
examined the Bd host–pathogen system in tropical Africa.
Bd was detected at all investigated sites, indicating that it is
ubiquitous in the sampled habitat types and may be wide-
spread in tropical Africa. We furthermore found that the
prevalence and intensity of Bd infection correlated with the
taxonomic and climatic patterns in the sampled region.
Our results suggest that there is a taxonomic component
to Bd susceptibility in the anuran communities studied.
Susceptibility appeared to be highest in the genus Amietia
(Ranidae), which consistently showed a high prevalence and
parasite load. The genera Ptychadena (Ptychadenidae) and
Hyperolius (Hyperoliidae) exhibited intermediate infection
levels and members of the genus Xenopus (Pipidae) were
seldom infected. Interestingly, the median parasite load of
infected individuals was not taxonomically biased, indicat-
ing that the extreme ends of the intensity distribution
constitute the difference.
During our survey, despite extensive collection efforts, no
dead or moribund anurans were encountered, nor were any
clinical symptoms of chytridiomycosis observed. Even the
most heavily infected individuals carrying a parasite load of
over one million GE appeared to be in a good body
condition. The high prevalence of Bd and the large number
of sub-clinically infected individuals may suggest that the
pathogen is enzootic in and possibly native to the studied
region. However, considering the cryptic disease progres-
sion characteristic for chytridiomycosis, it is not possible to
exclude that it may be causing mortality in the studied
populations undetected. Sub-clinical infections can build
up to a threshold, with symptoms only occurring in the
terminal phase of disease (see e.g. Carey et al., 2006; Voyles
et al., 2007), and field observations equivalent to ours have
even been made just before mass die-offs (Woodhams et al.,
2007). Furthermore, an enzootic state of Bd may be a
consequence of populations being in a post-decline phase
and that declines were merely not observed as they hap-
pened. There are now numerous examples of enzootic Bd
infection in amphibian populations globally (Retallick,
McCallum & Speare, 2004; McDonald et al., 2005; Long-
core et al., 2007; Brem & Lips, 2008; Woodhams et al., 2008;
Padgett-Flohr & Hopkins, 2009), and recent compelling
evidence suggests that Bd can have a marked impact even
c 2009 The Zoological Society of London 3
Amphibian chytrid fungus in Kenya J. Kielgast et al.

Table 1 Summary statistics for the distribution of Batrachochytrium dendrobatidis (Bd) across localities (see Fig. 1, Appendix S1) and genera
surveyed
Specimens Variance/mean
sampled Bd positive Prevalence (CI)a Max GE Mean GE Median GE ratio
Locality
Aberdares National Park (moorlands) 31.00 22.00 0.71 (0.53–0.84) 1 003 737.00 95 146.00 3966.00 739 406.00
Aberdares National Park (Salient) 111.00 43.00 0.39 (0.30–0.48) 120 350.00 7144.00 48.00 73 771.00
Kakamega Forest National Park 72.00 12.00 0.17 (0.10–0.27) 52 232.00 6059.00 96.00 40 414.00
Mt. Elgon National Park 11.00 5.00 0.46 (0.20–0.74) 14 610.00 2938.00 6.00 14 512.00
Nairobi (Karens) 109.00 27.00 0.25 (0.17–0.34) 48 945.00 2543.00 12.00 39 526.00
Saiwa Swamp National Park 126.00 62.00 0.49 (0.40–0.58) 227 779.00 5145.00 24.00 176 607.00
Shimba Hills National Park 150.00 6.00 0.04 (0.02–0.09) 9.00 6.00 6.00 7.00
Taita Hills (Mwatate) 21.00 4.00 0.19 (0.07–0.40) 980.00 296.00 101.00 828.00
Taita Hills (Mwundanyi) 116.00 34.00 0.29 (0.21–0.38) 10 395.00 587.00 7.00 6185.00
Thompson Falls 71.00 47.00 0.66 (0.54–0.76) 20 336.00 1124.00 65.00 9780.00
Tigoni Dam 50.00 9.00 0.18 (0.09–0.31) 565.00 83.00 12.00 443.00
Genus
Afrixalus (Hyperoliidae) 18.00 2.00 0.11 (0.02–0.33) 4.00 4.00 4.00 3.00
Amietia (Ranidae) 81.00 49.00 0.61 (0.49–0.71) 1 003 737.00 49 327.00 133.00 680 415.00
Amietophrynus (Bufonidae) 13.00 3.00 0.23 (0.07–0.52) 5.00 5.00 5.00 4.00
Hyperolius (Hyperoliidae) 418.00 180.00 0.43 (0.38–0.48) 120 350.00 2579.00 30.00 56 624.00
Kassina (Hyperoliidae) 30.00 1.00 0.03 (0.00–0.18) 2.00 2.00 2.00 2.00
Leptopelis (Arthroleptidae) 20.00 0.00 0.00 (0.00–0.17) 0.00 0.00 0.00 0.00
Phrynobatrachus (Phrynobatrachidae) 27.00 1.00 0.04 (0.00–0.18) 200.00 200.00 200.00 200.00
Ptychadena (Ptychadenidae) 126.00 31.00 0.25 (0.18–0.33) 48 945.00 2216.00 15.00 37 812.00
Xenopus (Pipidae) 128.00 4.00 0.03 (0.01–0.08) 18.00 7.00 4.00 14.00
a
95% confidence intervals on Bd prevalence in parentheses were constructed using Sterne’s exact method (Reiczigel, 2003).
CI, confidence intervals; GE, genomic equivalents.

1 1
(a) (b)
0.8 0.8

0.6 0.6
Prevalence

Prevalence

0.4 0.4

0.2 0.2

0 0
0 5 10 15 20 15 20 25
−0.2 −0.2
Minimum temperature (°C) Minimum temperature (°C)

1 Figure 2 Simple linear regressions illustrating

1
(c) (d) the relationships between the mean Bd preva-
0.8 0.8 lence at the studied sites and (a) minimum
temperature (r2 = 0.825, Po0.001), (b) maxi-
0.6 0.6
mum temperature (r2 = 0.666, P= 0.002), (c)
Prevalence

Prevalence

0.4 0.4 precipitation (r2 = 0.002, P= 0.904) and (d) alti-

tude (r2 = 0.709, Po0.001); 95% confidence
0.2 0.2 intervals are indicated as vertical bars. The grey
lines represent the 95% confidence limits for
0 0
0 50 100 150 200 0 1000 2000 3000
the mean of the prediction of a given value
−0.2 −0.2 (dotted line) and the 95% confidence limits on a
Precipitation (mm) Altitude (m) single prediction for a given value (solid line).

decades after becoming enzootic (Murray et al., 2009). Existing accounts of Bd in African amphibians are
Hence, the apparently enzootic state of Bd found in the equivocal regarding pathogenicity and so far little data are
present study should be regarded as a conservation concern available. In tropical Africa, Bd has been reported to be
and requires further investigation. present in Nigeria (Imasuen et al., 2009), The Democratic

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J. Kielgast et al.
Republic of Congo (Greenbaum et al., 2008), Uganda
(Goldberg, Readel & Lee, 2007) and Tanzania (Weldon
et al., 2004), while a single study reports not detecting Bd in
Cameroon (Doherty-Bone et al., 2008). In Tanzania, Bd was
suggested to be linked to the decline and disappearance of
the Kihansi spray toad Nectophrynoides asperginis, sup-
ported by finding two out of four dead individuals collected
during the decline to be Bd positive (Lee et al., 2006). The
species was categorized under ‘rapid enigmatic declines’ in
the IUCN Global Amphibian Assessment 2004 (Stuart
et al., 2004), which have been suggested to cover over Bd-
driven extinction processes (Skerratt et al., 2007). However,
the Kihansi spray toad was endemic to a single waterfall
system (c. 2 ha.) that was radically changed by the construc-
tion of a power plant before declines (Krajick, 2006). The
aetiology of this decline is therefore highly confounded by
habitat change and the role of Bd is unclear. All other
studies from tropical Africa have not been coupled with
observations of mortality or decline. Goldberg et al. (2007)
conducted a single site survey with methodology similar to
ours and report congruent findings with 22% prevalence in
109 specimens and no mortality events noted in the region.
The remaining reports are accounts of small-scale sampling
and single Bd-positive individuals.
In the South African region, Bd has been found to be
widespread but no associated population declines have been
observed (Hopkins & Channing, 2003; Weldon, 2002; Wel-
don et al., 2004; Smith et al., 2007). However, Bd-associated
mortality in frogs has been detected twice without presum-
ing an aetiological link. The findings of the diagnostic
characteristics of chytridiomycosis (Lane, Weldon & Bing-
ham, 2003), and patterns of mortality and infection resem-
bling a die-off (Hopkins & Channing, 2003), call for further
investigations, but do not contradict an endemic presence of
Bd in the region.
Information on the susceptibility of African amphibians
to Bd under controlled conditions is limited to pipids and
indicates susceptibility in the western African species Silur-
ana tropicalis but resistance in Xenopus laevis (Reed et al.,
2000; Parker et al., 2002; Fisher & Garner, 2007). This has
not yet been verified by experimental infection challenges
but is solely based on observations of mortality and aclinical
infection in captive populations.
It has repeatedly been shown that the pathogenicity of
chytridiomycosis is context dependent and particularly
affected by environmental parameters, for example tem-
perature and precipitation (Bosch et al., 2001; Daszak,
Cunningham & Hyatt, 2003; Berger et al., 2004; Kriger &
Hero, 2006; Kriger, Peregolou & Hero, 2007; Kriger, 2009;
Longo, Burrowes & Joglar, in press). Furthermore, the
environmental suitability for a species at a given site may
affect the capacity of the host’s immune system affecting the
prevalence and intensity of Bd infections (Raffel et al., 2006;
Fisher, 2007). Our results indicate that prevalence decreases
with higher monthly minimum and maximum temperatures
(Fig. 2), thus supporting previous findings (i.e. Berger et al.,
2004; Piotrowski, Annis & Longcore, 2004; Kriger & Hero,
2006; Rödder, Veith & Lötters, 2008). In our study, pre-
Animal Conservation 13, Suppl.1 (2010) 1–8
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Amphibian chytrid fungus in Kenya
cipitation patterns did not correlate with Bd prevalence (Fig.
2). The species sampled were a priori the most abundant;
hence, it is understandable that species compositions varied
among sampling sites. The correlation detected between
temperature and prevalence may therefore either reflect
varying degrees of climatic suitability of the study sites for
Bd (e.g. coincident with the results shown by Piotrowski
et al., 2004; Kriger et al., 2007; Rödder et al., 2008) or
differences in the susceptibility of the sampled amphibian
communities or both.
The question remains open as to whether the Bd panzoo-
tic has its origin in Africa. If this is the case, the observed
patterns of high prevalence and apparently low virulence
found in the present study can be explained in a co-evolu-
tionary context, that is evolution of host resistance, aviru-
lence or attenuation of the local strain of Bd (McCallum,
2005). There is accumulating evidence that Bd exists in
various strains exhibiting markedly different levels of viru-
lence (Berger et al., 2005; Retallick & Miera, 2007; Fisher
et al., 2009) although recent evidence points towards no
correspondence between the genetic lineage of strain and
virulence (James et al., 2009). Comparative clinical studies
investigating host–pathogen dynamics by experimental ex-
posure of African and non-African amphibian hosts and Bd
isolates may be a way forward in testing the ‘out-of-Africa’
hypothesis. Here, the conditional nature of pathogenicity
should be evaluated, by including a range of climatic
conditions, to determine the relative importance of the
intrinsic taxonomic components of susceptibility and cli-
matic factors. Moreover, the ‘out-of-Africa’ hypothesis
needs to be addressed by analysing the phylogenetic patterns
and global genetic diversity in Bd including a broader
coverage of Bd isolates from Africa than are currently
available (James et al., 2009).
It should be underlined that disease surveys as such have
a limited capability of detecting declines and proving patho-
logical causality. This requires clinical experimental studies
of host–pathogen dynamics (including fulfilment of Koch’s
postulates) and long-term disease surveillance coupled with
studies of host population dynamics. There is an urgent need
to improve our knowledge of the conservation status of
tropical Africa’s amphibian fauna. A crucial step towards
this is a qualified assessment of the risk imposed by wide-
spread Bd infections found in the present study. As a
priority, we urge the initiation of population monitoring
focusing on susceptible amphibian communities in the high-
lands of tropical Africa.
Acknowledgements
We are grateful to Trent Garner and Andrew Cunningham
for making available the facilities and expertise of the
Institute of Zoology, London. This extraordinary helpful-
ness was completely essential in realizing the present study.
We thank The National Museums of Kenya and staff for
fruitful cooperation. Susanne Schick, Felix Müller, George
Kennedy and Beryl Bwong were of invaluable assistance
c 2009 The Zoological Society of London 5
Amphibian chytrid fungus in Kenya
during fieldwork. The first author would like to express
deepest gratitude to Mads Frost Bertelsen and Copenhagen
Zoo for decisively contributing to the study and Mogens
Andersen and Peter Gravlund for practical support. Finally,
we would like to extend many thanks to two anonymous
reviewers whose helpful comments greatly improved the
paper. The project was financed through the Copenhagen
Zoo/Center for Zoo and Wild Animal Health, The Danish
WWF and Aase og Ejnar Danielsens Fond, BIOLOG-
BIOTA from the Federal Ministry of Education and Re-
search (BMB+F, Germany), H.R. Frederiksen og Grete
Siim Frederiksen Fond, Fonden Kjebi. D.R. is grateful to
the ‘Graduiertenförderung des Landes Nordrhein-Westfa-
len’ for financial support. The Kenya Wildlife Service,
KWS, kindly granted permission for this work (No. KWS/
RP/5001).
Conflicts of interest: None.
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Supporting Information
Additional Supporting Information may be found in the
online version of this article:
Appendix S1. Localities sampled including altitude in m
a.s.l. (see Fig. 1).
Appendix S2. Bd distribution within the sampled am-
phibian communities (species identification after Channing
& Howell, 2006). Exact Clopper-Pearson 95% Confidence
intervals are presented for the prevalence. Parasite load
summary statistics are based on the infected fraction of the
sample.
As a service to our authors and readers, this journal
provides supporting information supplied by the authors.
Such materials are peer-reviewed and may be re-organized
for online delivery, but are not copy-edited or typeset.
Technical support issues arising from supporting informa-
tion (other than missing files) should be addressed to the
authors.
c 2009 The Zoological Society of London