Attachement 1
A draft of the multidisciplinary research project
Evolutionary biogeography of fruit bat, Rousettus
aegyptiacus, in the Mediterranean region.
GAAV grant IAA601110905
02 Summary
In geographic and genetic structure, the Mediterranean population of fruit bat, Rousettus
aegyptiacus, an only offshot of the family Pteropodidae beyond tropes, markedly differs from
all other bats of that region. The project is intended to explain the background factors of
these specificities, history and life-history traits promoting colonization of the Mediterranean
and maintenance of local populations and answer which aspects of the climatic and
environmental history of the region affected it essentially. The project combines methods of
fine-grain molecular phylogeography and a long-term field study of the biology of the model
species in several geographically distant model subpopulations and is expected to provide a
complex information on the Mediterranean fruit bat including estimates of its total
abundance and a critical reexamination of its biogeographic specificities.
03 Principal investigator
Charles University in Prague
Persons working on project:
Prof. RNDr. Ivan Horáček, Csc: horacek@natur.cuni.cz
Ing. Petr Jedlicka Ph.D.
Mgr., Tomáš Bartonicka Ph.D.: bartonic@sci.muni.cz
RNDr. Petr Benda, PhD
Doc.RNDr. Vladimír Hanák
RNDr. Pavel Hulva, PhD: hulva@natur.cuni.cz
Mgr.Helena Jahelkova, PhD : hjahel@yahoo.com
Mgr.Radek Lucan : rlucan@centrum.cz
1. The Problem
Biogeography, an influential integrative platform of last decades for various disciplines of
organismic biology (Lomolimo et al. 2006) suffers from obvious incompatibility of the
phenomena and explanations revealing at different scales (Storch et al. 2007). The factors
sufficiently explaining specificities of a range and its coverage at a large scale have often
little to do with those responsible for same phenomena at the local scales.
Consequently, the task of the day is to precise how the taxon-specific setting of life history
traits, their performace under limiting effects of local conditions and aspects of the local
history influence the current form of its distribution and its local dynamics.
The aspects of the Mediterranean range of the Egyptian Fruit bat, Rousettus aegyptiacus,
seem to be a particularly suitable model for such studies. The species occupies a very large
range coveringconsiderbale part of Africa from the Cape Region to Egypt (Bergmans 1994,
Kwiecinski and Griffiths 1999). The extension of its range into the Eastern Mediterranean, to
which the project is concentrated, represents for several reasons a stricking biogeographic
phenomenon which is not well understood and calls for a detailed explanation.
It is the only off-shot of the range of family Pteropodidae beyond the tropical zone
(Simmons 2006)and the reasons why it is situated just there and what promoted there the
species survival are largerly unknown. R.aegyptiacus is the only frugivorous element in the
bat fauna of the Palearctic region (Horácek et al. 2000), and also in other respects it shows
several remarkable differences from other Palearctic bats. Its Palearctic range is restricted
onto a narow strip of the thermo-Mediterranean zone (in sense of Blondel and Arison 2001)
and splitted there into several more or less isolated subpopulations (SE Turkey, Cyprus, W
Syria, Lebanon to Israel, Nile valley in Egypt and Sinai). It absents from all islands except
for Cyprus (which is the least easily colonizable island of the region - cf. Benda et al. 2007).
The comprehensive analysis of all chiropteran records from the Eastern Mediterranean (as
summarized by Benda and Horácek 1998; Hanák et al. 2001;Benda et al. 2003, 2006, 2007;
Horácek et al. 2007a, b) revealed for this species an extreme disproportion between high
mean frequency (1.09) and very low percentage of actually occupied cells (0.16) in the
database of 5869 bat records from the Eastern Mediterranaen (26 to 42 deg N,
18 to 60 deg E, expressed in the 2x2 deg grid system - Horácek et al. 2007a).
Against expectancy, preliminary results of genetic screening of the Mediterranean and Middle
East populations of the species (Hulva et al. in prep.) revealed a stricking genetic
homogeneity throughout all the region (cyt b, genetic distances from 0.001 to 0.016) though
shalow signals indicating a marked phylogeographic patterning do exist here as well. The
colonization history and the relationship among particular local populations are completely
unknown, of course, similarly as the behavioral dispositions of the species to expected
dispersal events which could increase a gene flow rate. Anecdotical data available on that
respect (mostly Makin 1990) suggests migrations between roosting and feeding sites up to
13 kms, and interseasonal migrations up to the distance of 30 and 80 kms, respectivelly.
Nevertheless, it is completely unknown to which degree the respective spatial dynamics is a
common feature of the species, which is seasonal turnover in composition of its colonies and
roost occupancy etc. Surprisingly, except for few studies dealing mostly with diet and
reproduction (Kulzer 1958, 1979, Makin 1990, Korine et al. 1994, 1999, 2004) almost no
comprehensive data are available on other aspect of biology and life history traits of the
species or the information is restricted onto observations on captive bats. Among other it
concerns a structure of colonies, its seasonal variation (including expected sex seggregation
and seasonal migrations) and also reproductive pattern that seems to be biomodal in Israel
and Cyprus (Spitzenberger 1979, Makin 1990, Korine at al. 2004) but may vary throughout
the region as suggested by our observations (Benda et al. 2007).
Rousettus was often considered an agricultural pest in the region and subjected to extensive
erradications (Makin 1989, 1990, Hadjisterkotis 2006). Of course, it is undoubtedly quite a
unique element of the Palearctic fauna (and even that of the European Union – cf. Cyprus)
which should be subjected to a very rigorous conservation interest. Unfortunately, reliable
scientific arguments enabling to establish a proper balance between the controversial views
still absent. The present project arising of the solid platform of our previous results
concerning the distribution and biology of the species in Eastern Mediterranean (cf. Benda et
al. 2006,2007) is expected to fill the gap in all the above mentioned respects.
2. Objectives of the project
The project is intended to explain the background factors responsible for specificities of the
Mediterranean range of R.aegyptiacus, reconstruct its history and explain which of its life
history traits play a role of key mechanisms of the range colonization and maintenance of
localpopulations.
We expect to respond that task via answering the following particular questions:
(1) Does the E-Mediterranean range of the species consists of more or less separated set
centers oflocal aggregation or such a view is mere a result of incomplete record?
(2) Which are actual abundances of the species in particular centers of aggregation and what
would be a total population of the species in its Mediterranean range?
(3) Which are molecular divergences among particular subpopulations and which the
molecular datings of them?
(4) Are there the divergences of local populations accompanied with differences in their
phenotypic and/or life-history traits?
(5) Which is migration capacity of the species and which are actual home ranges, spatial
dynamics of particular colonies and seasonal variation in it?
(6) Which is social structure of a colony, its social cohesiveness, and differences between
particular age cohorts and sexes in degree of filopatry and in dispersal patterns?
(7) How does R.aegyptiacus respond to restriction of food resources in winter?
(8) Which are actual forms of the interdependence with carob (Ceratonia siliqua) as its
ultimate winter diet (cf. Benda et al. 2007) and their role in range history and dispersal?
(8) Are there differences among local populations in these traits, and if so: which are further
correlates of these differences and which is their geographic variation? Do they show
divergence-by-distance patterns or scaling with climatic and environmental currents?
(10) Which were forms of influence by human population in historical past (including
anthropogenic spread of carob and other food resources) and their possible impacts upon
the species under study?
We are quite convinced that all these questions can be completely answered with aid of a
field investigations combining different methods and approaches and focused on a long-term
study in carefully selected model populations. The outputs of these field studies obligatory
supplemented with results of molecular analyses (from individual identifications, haplotype
and microsatellite mapping to phylogeograhic reconstructions and molecular clock datings)
will undoubtedly provide a reliable basis for a synthesis responding to major project
objectives as drawn above.
3. Time schedule
The project is scheduled for five years. The long span of the project is a necessary
prerequisite for filtering effects of temporal variation and obtaing relevant data on
population dynamics, stability of colonies, and state of particular life-history traits under
study.
The first year: field expeditions in the region aimed to refining available distributional record,
establishing the conditions for a long-term investigation (including model populations and
colonies) and field testing of reliability of instrumental equipement and methods (marking,
radio-tracking, recaptures, non-invasive DNA sampling). Finishing of the molecular analyses
of the samples obtained prior beginning of the project (DNA isolation, PCR, sequencing),
screening of variability of nuclear markers, testing microsatellite primers. Analyses of
phenotypic variation.
From the second year onwards: the regular sampling, observations and radio-tracking
studies in model populations will be pereformed with repeated yearly controls both in
summer and winter period. At the same time the routine molecular analyses of field samples
will be continually performed. Supplementary studies on phenotypic variation.
The third year: Finishing studies on distribution, phenotypic variation and large-scale
phylogeography, presenting manuscripts on these subjects. The fourth and fifth year of the
project will be focused to synthesis of particular outputs, statistical analyses and presenting
manuscripts concerning life-history traits and aspects of evolutionary biogeography.
4. Significance of the project, expected public interests and outputs
The scientific outputs of the project will provide detailed information of fine scale
biogeograophy and biology of the model species and supposedly will demonstrate
possibilities of integration of various approaches in organismic biology and the way to cross
constitutional incompatibility of the global and local scale biogeography.
The detailed information on distributional statut of one of the most interesting elements of
the West Palearctic mammal fauna, abundance and biology of its local populations in the
Mediterranean region will appreciated also in other contexts. These informations are
expected to be of a considerable international interest as a basis for the EU conservation
strategy focused on the species and at the same time as a reliable source of arguments to
discussions on population management and control of that species forced by local
authorities. Last but not least, integration of students in the project will contribute to
university education and the new techniques (e.g. automatic radiotelemetry system) and
approaches that are to be applied in frame of the project could be worth of a common
interest as well. The project should results in a series of journal publications, conference
presentations and in a scientific monograph besides a detailed report on the species status
for EU Conservation authorities.
5. Conceptual setting and methods
The project expects a considerable amount of
(i)
field investigations in the regions (with particular focus to model populations in
Cyprus, Lebanon, Turkey and Sinai),
(ii)
laboratory studies (mostly molecular and phylogenetic analyses) and
(iii)
post-hoc investigations (statistical data analyses,GIS analyses of radio-tracking
data, testing effects of climatic and environmental variables etc.).
5.1. Distributional status and abundances of local populations
The database of all known records of the species in the Eastern Mediterranean gathered in
the previous stage of study (comp. Benda et al. 2006, 2007, Horácek et al. 2007) will be
subsequently enlarged with the records obtained by systematic search for its roost and
foraging ground in major centres of its distribution (Lebanon, Cyprus, Turkey, Nile valley,
Sinai) as well as in the areas between them from where only exceptional records are
available. In the selected model areas we will attempt to discover all the sites used by the
species and establish a direct estimate of its actual abundance. In the case of R.aegyptiacus,
such a task, quite illusory for any other bats, can be actually accomplished: it is a large,
distinguishable and highly social animal demanding spacious underground roosts which
availability in limited. The occupied roosts are locally well known as a rule what further
increase the chance to discover them completely. The direct abundance estimations will be
confronted with the probabilistic estimates obtained by a marking-recapture
techniques. Based on resulting data and detailed distributional information we will model a
series of alternative hypotheses on the total population of the species in the region.
5.2. Sampling techniques, handling and individual identification The bats will be captured
with aid of mist netting and/or with potrable Tuttle's trap either in entrances of roosts, at
night roosts or at foraging grounds. All captured bats will be sexed, weighted and measured
(FA), their individual age will be estimated by degree of tooth wear and
reproduction conditions based on the standard external characters (nipples type. milk
production, testes etc.). The samples for DNA analysis (and subsequent gentic individual
identification) will be taken from every captured individual. A swab of oral mucose is
proposed as a method to be applied in routine handling. At the first stage of the project,
tThe standard control sampling by sterile biopsy from the plagiopatagium will be applied as a
parallel control. In the model colonies selected for a long-term study we will apply two
techniques of individual marking: chain ball necklaces with numbered rings (Bumrungsri et
al. 2007) and passive RFID transponders (supposedly BMDS IMI-1000 or Assion IDD 101)
enabling individual identification either by handling (with aid of iMAx Black Label Microchip
Scanner), directly in bat clusters in their roosts or just at roost entrance via
automatical recording (comp. Dell'omo et al. 1997).
5.3. Molecular methods
Molecular part of the project will cover topics of (a) large-scale phylogenetic and
phylogeographic structure of the species under study and dating respective divergences, (b)
geography of particular haplotypes and microsatellites, (c) genetic structure of particular
colonies and subpopulations and their mutual relations, and (d) molecular techniques of
individual identification. Some of the techniques were apllied already in previous stage
project and a large still not completely analyzed material collected in the region (about 160
ind.) is already available.
Approach: Initially, most studies of bat phylogeography were performed with single locus
attempt, enabling proposal of a posteriori hypotheses in terms of divergence and gene flow
among populations. Recently, a multilocus approach (including mitochondrial and nuclear
genes) fortifies testing of a priori hypotheses in terms of coalescent permit through the
estimation of demographic parameters, however, phylogeographic studies on bats using
nuclear data are relatively scarce. The screening of single nucleotide polymorphisms in large
scale, usual in traditional mtDNA attempt, is now increasingly available also for nuclear
markers (e.g. Brumfield et al., 2003). Although genealogical trees derived from recombining
haplotypes are conceptually different from recombinant ones (e.g. Hare 2001), the use of
both mt and nuc DNA enables interpretation in terms of different characteristics of both gene
pools. For instance, the haploid matter of mtDNA results in increased sensitivity to reduction
of population size, causing easier sorting of mitotypes during bottlenecks (e.g. Seddon et al.
2001). The maternal heredity of mtDNA (respectively paternal heredity of Y chromosome)
could be used in assessing of sex-biased contribution to gene flow. Female philopatry typical
for mammals (e.g. Miller-Butterworth et al., 2003) could be in some cases substituted by
different pattern of dispersal. We want to interpret comparison of mt and nuc DNA results in
terms of population parameters (lineage sorting, character of post glacial expansions,
bottleneck/ size of founding populations, effect of repeated contraction to refugia etc.).
In the first part of the project, we will choose appropriate genetic markers. We will continue
with analysis of mitochondrial genes (e.g. Hulva et al., 2004). We will perform screening of
variability of nuclear markers and make the final choice according to the results. Several
useful nuclear genes were amplified in the order Chiroptera (e.g. Teeling et al., 2002; 2006;
Stadelmann, 2007). Using of Y chromosome (enabling assessment of paternal contribution
to gene flow) and X chromosome markers (combining haploid state in males vs. diploid in
females) will be considered based on the detected variability. Extensive screening of sex
chromosome markers in mammals with good results for the order Chiroptera was published
(Hellborg and Ellergen, 2003; 2004) and used in phylogeographic studies in rodents (e.g.
Hellborg et al., 2005). We will perform also crosspecies analysis in order to test
microsatellite primers that were developed for species from the family Pteropodidae
(Rousettus leschenaulti, genus Cynopterus, Pteropus conspicillatus - Hua et al., 2006;
Storz, 2000; Fox et al., 2007). After sufficient amount of microsatellite loci with suitable
variability will be obtained, we will use microsatellites as additional markers for individual
identifications and characterization of population structure of Rousettus aegyptiacus, and
relations among particular local populations.
The tissue samples will be obtained either by sterile biopsy from the chiropatagium, the
minimum invasive method developed especially for field studies of bats (Wortington, Wilmer
and Barratt 1996), or (after testing its reliability) by a swab of oral mucose which will further
minimize stress of handling. The samples will be stored in 95% ethanol.
DNA analyses: The genomic DNA will be extracted from alcohol preserved or dry tissue
samples from museum collections or from biopsies (Wortington-Wilmer and Barratt 1996) by
the Dneasy Blood and Tissue Kit (Qiagen). Older museum specimens will be lysed in
proteinase K + Chelex (Biorad) solution and the DNA from supernatant will be purified using
QIA Quick PCR Purification Kit (degraded DNA removal method; Spitzenberger et al., 2001).
Selected loci will be amplified by PCR, amplicons will be excised from agarose gel, purified
with the QIAquick Gel Extraction Kit (Qiagen) and sequenced by BigDye Cycle Sequencing
Kit (ABI PRISM; with the same primers as PCR) and automated DNA sequencer (PE3100
Avant Genetic Analyzer) at the home university or under contract (Macrogen).
Chromatograms will be edited and resultant sequences aligned (Chromas; SeqMan;
ClustalW).
Data analyses and reconstructions of phylogeny: Obtained datasets will be combined with
published data and tested for phylogenetic information content - standard g1 statistic for
measuring the skewness of tree lengths of alternative trees (Hillis and Huelsenbeck, 1992),
saturation tests (e.g. Xia et al. 2003, Daugbjerg & Andersen 1997). Standard methods of
molecular phylogenetics will be used for creating and testing phylogenetic hypotheses
(distance analysis; maximum parsimony – simple, generalized and weighted; analysis of
sequence evolution model; maximum likelihood; bootstrap; Bayesian analysis; posterior
probabilities; spectral analysis; molecular clock). The appropriate software will be used –
PAUP, Modeltest, MrBayes, Spectrum, PHYLIP etc. In order to examine the history of
analyzed populations in detail (size changes, range expansion in case of recolonization
Europe, gene flow level etc.) the coalescent theory and phylogeography approach will be
used (mismatch distribution, lineages through time, skyline plots,
nested clade analysis, etc. – TCS, Arlequin, GeoDis etc).
5.4. Life-history traits
We expect a large amount of capture-recapture data sufficient not only for analysis of spatial
behaviour (fidelity, filopatry, migrality etc.) but also for statistical estimation of various
demographic variables, including survival curves, and – last but not least – for stratified
estimations of population abundances (with aid of marking-recapture techniques). The
characteristics of reprodution and its annual course in particular populations will be obtained
by repeated controls in model colonies (daily for a period of several weeks, both in summer
and winter months) and statistical analyses of reproductive data in captured individuals.
5.5. Social structure of a colonies, social cohesiveness, roost switching and roost
fidelity
We aim to understand the evolution of complex social behaviour as a function of relatedness
among reproducing and non-reproducing females. We hope that our data will improve the
general understanding of extended families, and allow us to evaluate models of reproductive
share in animal societies. Further we will test if maternity colonies are closed societies,
comprising closely related and genetically unrelated females that live together for many
years; if model colonies regularly split into several subgroups that occupy different roosts
and how often individuals switch roosts. Using different techniques of individual marking
(chain ball necklaces and passive RFID transponders with microchip scanners) in three
model underground roosts, where the most individuals will be marked, help us to detect the
social organization of maternity colonies. the fact if the females need familiar co-operation
partners for raising their young. Further we will research into a probably profit from sociality
because of warming by non-reproductive females, babysitting and information transfer
about roosts or foraging areas.
5.6. Analysis of spatial dynamics with aid of an automatized radiotracking system
Standard techniques of radio-tracking (with aid of Holohil PD-2 3.8 g transmitters lasting for
6 months, AOR Inc. AR8200 receivers and threee-element Yagi antenae) will be used for
long-term tracking of spatial activity of selected individuals including roosts alternations.
Besides that we will apply the technique of automatic radio-tracking recently developed (and
succesfully tested in field) by T.Bartonicka in cooperation with P.Jedlicka., Institute of
Scientific Instruments of the ASCR, v.v.i. Brno. Within this proposed project a set of tracking
units wil be designed on the bases of previous experience with respect to the needs of this
specific application. The technique enables simultaneous tracking of 50 individuals and will
be applied in repeated study of spatial activity and dispersal in several model colonies. The
system is based on stationary stations operating with Doppler Radio Direction Finding which
automatically monitor up to 50 different frequencies, continually store their position in
memory of a collector unit (via GSM modem) and express them in GIS coordinates via
triangulation from inputs of individual stations.
The data are exported in ArcView compatible format and will be further analyzed with aid of
Animal Movement extension via standard MCP and Kernel estimations. The field tests
confirmed reliability of the instrumental equipement, variation of a directional field
below 5 deg at a single station setting and a accuracy of position identifiaction exceeding
those obtained by standard methods. Standard radiotracking research will be performed by
up to four mobile researchers, fixes of followed animals will be coordinated by walkie-talkies
and tracked using GPS device. Once a feeding site is located, its position will be maped and
precise information on type and density of food supply will also be described during
subsequent days.
5.7. Foraging biology
The data arising from radio-tracking studies will be analysed with particular respect to
spatial distribution, size and distances of foraging grounds and differences in these variables
between sexes, age cohorts, and particular periods of the year. Proper analytic software
tools (GIS, Homerange etc.) will be applied on data to obtain detail information on spatial
behaviour of hunting bats. The data on spatial activity will be analyzed using multimethod
approaches, including compositional analysis. This technique makes full use of time budget
data, and by using the individual animal as the sampling unit, avoids problems of
autocorrelation of data points that are inherent in methods such as chi-square and
simultaneous confidence intervals on pooled data (Neu et al. 1974, Aebischer and Robertson
1992, Aebischer et al. 1993). Besides that we will map the spatial distribution of plant
species known to be consumed by fruit bats in the surroundings of the model roosts to
assess relation of position of particular roosts to distance and distribution pattern of food
resources. Following methods by Korine et al. (1999), we will collect food remnants and
faeces of fruit bats in their day- and night- roosts in different periods of the year. Plant
pieces and seeds will be determined using reference colection of species occuring
in particular study area. Based on these data, we will describe year round changes in the
diet and test the obvious interregional differences suggested by literary data (comp. e.g.
Albayrak et al. 2008).
Particular attention will be paid to winter diet and forms of interactions with Ceratonia
siliqua, modes of its harvesting and handling by bats (which represent most probably the
major agent ofits seed disperal). Intensity of harvesting during course of winter will be
studied by daily controls ofmodel trees and direct night observations.
We expect to obtain in the compared model populations the comprehensive answer to the
questions like: How does change food supply, its spatial ditribution and its use by fruit bats
duringthe season? Is there any rule for how do fruit bats exploit food resources? How does
change theproportion of native and introduced (i. e. mostly commercial) plant species in the
diet of fruit bats with respect to season of the year and geographic area? How do fruit bats
cope with severe conditions during winter period when almost no food resources are
available – do they rely on accumulating fat reserves or, alternativelly, change their spatial
activity and/or feeding habits?
5.8. Synthesis of the results
The outputs of the above mentioned particular approaches will be summarized into complex
characteristics of individual model populations. The differences between populations in
particular variables will be examined and their relations to abundance characteristics,
climatic and environmental currents and genetic specificities of the respective populations
will be analyzed with aid of various multivariate techniques. The possible effects responsible
for the observed pattern of local distribution will be formulated and discussed in context of
the historical and ecological biogeography of the region.
6. Cooperation with foreign institutions
The field study will be effected in close cooperation with local specialist and local authorities
(Prof.R.Sadek, Dr.M.Abi-Said, Rena Karanouh and Speleo-Club Liban in Lebanon, Dr.
E.Hadjisterkotis, and P.Georgiakakis in Cyprus, Prof. I.Albayrak, Prof.A.Karatas in Turkey).
The comparative studies of museum material will be underataken in Natur-Museum u.
Forschunginstitut Senckenberg, Frankfurt a.M., BRD, Naturhistorisches Museum Wien and
Natural History Museum London.
7. Competency of the applicants and their contribution to the project
Ivan Horácek, Vladimír Hanák and Petr Benda (Charles University, Prague), the senior
scientist included in the project, are prominent specialists in biogeography, taxonomy and
ecology of bats, all with extensive experiences in study of Mediterranean bats (including
authorships of a mongraphic series Bats of Eastern Mediterranean. Part 1-7 covering more
than 700 pp.) and with a considerable foreign reputation. They all will take part in field
studies, and are responsible for phenotypic and chorologic analyses. The applicant,
I.Horácek is further responsible for coordination of particular aspects of the project.
Radek Lucan, a PhD student of I.Horácek, who is expected to receive a full employment
from the grant, will be responsible for major bulk of field studies. He is extrardinary
experienced in field study of bats, both in long-term investigation of autecology of various
Palearctic species and in field work in diversed exotic regions, including all the countries of
the Eastern Mediterranean.
Tomáš Bartonicka, assistant professor at Institute of Botany and Zoology MU Brno, is a
specialist in long-term field investigation of roosting and foraging biology of bats and in
related instrumental techniques. He essentially contributed to development of the technique
and instrumental equipment of automatized telementry that will be applied in field study of
spatial activity.
Petr Jedlicka, research assistent at Institute of Scientific Instruments ASCR Brno, v.v.i.
(Academy of Sciences of the Czech Republic, is a technical specialist contributed to the
optimalization of automatized telemetry system and transmision of tracking data into GIS
software.
Pavel Hulva, will be responsible for coordination of the project and most of the laboratory
investigation. He has considerable practical experience in molecular genetics and
evolutionary disciplines and a series of valuable achievements in that branch including first
molecular data on the family Craseonycteridae (Hulva and Horácek, 2002), phylogeographic
analysis of the family Rhinopomatidae (Hulva et al., 2007), discovery of cryptic species
diversity in the Mediterranean populations of P. pipistrellus group (Benda et al., 2003a;
Hulva et al., 2004; Benda et al., 2004a; Hulva et al., 2007). He also took part in field
expeditions to Iran, Cyprus, Crete, Lebanon and Sicily.
Peter Vallo, a PhD student in Comenius University, Bratislava, will take part in molecular
analyses, in particular in microsatellite studies and routine analyses of individual identity.
We expect that one or two MSc students will joint the project and assist both in field and
laboratory.
Helena Jahelkova, research assistent in Charles University, will be responsible for the field
work on ethology, acoustic communication and part of foraging ecology.
8. Technical and instrumental background
The field study in the region will partly profit of the facilities of the cooperating local
institutions,nevertheless, major bulk of expenses is expected to be covered by the grant.
Among other this will include car rentings, expenses of installation of stations of automatized
radiotracking system and living expenses of the investigators. Consequently, the travel
expenses represent a considerable part of the project budget.
The proper instrumental equipment is mostly already at our disposal, microchip
transponders, transmitters and additional receivers will be purchased.
Molecular analyses will be performed in the Laboratory for Biodiversity Research at
Department of Zoology, Faculty of Science, Charles University. Sequencing will be partly
performed under contract (Center of service laboratories at Charles University, Macrogen).
Considering the specialization, the home laboratory focuses on isolation and analyses of
biopolymers (with focus of production of sequence data). Nevertheless, the laboratory is
equipped for performing complete genetic engineering projects and thus has capacity for
testing proximal mechanisms when necessary.
Materials and chemicals are either at disposal or will be provided from resources of the
grant.
9. References
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Rousettusaegyptiacus, in Turkey (Mammalia: Chiroptera). Turk.J.Zool. 3211-18.
Aldridge H.D.J.N., Brigham R.M. 1988. Load carrying and manoeuvrability in insectivorous
bats: atest of 5% “rule” of radio– telemetry. Journal of Mammalogy, 69: 379–382.
Aebischer N. J., Robertson P. A. 1992. Practical aspects of compositional analysis as applied
topheasant habitat utilization. In: Priede, I. G. and Swift, S. M. (eds), Wildlife telemetry:
remotemonitoring and tracking of animals. Ellis Horwood, Chischester, UK, pp. 285-293.
Aebischer, N. J.,Robertson P.A., Kenward R.E. 1993. Compositional analysis of habitat use
fromanimal radio-tracking data. Ecology 74: 1313-1325.
Benda P., Horácek I. 1998. Bats (Mammalia: Chiroptera) of the Eastern Mediterranean. Part
1.Review of distribution and taxonomy of bats in Turkey. Acta Societatis Zoologicae
Bohemicae 62,255–313.
Benda, P., Hulva, P., Andreas, M., Uhrin, M. 2003a. Notes on the distribution of Pipistrellus
pipistrellus complex in the Eastern Mediterranean: First records of P. pipistrellus for Syria
and of P.pygmaeus for Turkey. Vespertilio 7, 87–95.
Benda P., Ivanova T., Horácek I., Hanák V., Cervený J., Gaisler J., Gueorguieva A., Petrov
B.,Vohralík V. 2003b. Bats (Mammalia: Chiroptera) of the Eastern Mediterranean. Part 3.
Review ofbat distribution in Bulgaria. Acta Societatis Zoologicae Bohemicae 67, 245–356.
Benda P., Hulva, P., Gaisler, J. 2004a. Systematic status of African populations of Pipistrellus
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