| Abstract | Divokoza (Rupicapra spp.) rasprostranjena je diljem Europe gdje nastanjuje srednje i visoke
planinske regije, a u iznimnim slučajevima nizinske predjele riječnih kanjona i područja na
razini mora. Danas prema morfološkim i bihevioralnim svojstvima postoje dvije vrste
divokoza: južna divokoza Rupicapra pyrenaica (s podvrstama parva, pyrenaica i ornata) i
sjeverna divokoza Rupicapra rupicapra (s podvrstama cartusiana, rupicapra, tatrica,
carpatica, balcanica, asiatica i caucasica). Dvije od sedam podvrsta sjevernih divokoza (R.
r. rupicapra i R. r. balcanica) nastanjene su u Hrvatskoj, dok je balkanska divokoza
rasprostranjena diljem Balkanskog poluotoka još u osam zemalja: Bosna i Hercegovina,
Srbija, Crna Gora, Kosovo, Sjeverna Makedonija, Albanija, Bugarska i Grčka.
Trenutno znanje o genetskoj raznolikosti i strukturi populacija balkanske divokoze
ograničeno je na lokalna istraživanja. U ovom istraživanju korištena je kombinacija seta
mikrosatelitnih lokusa i sekvence kontrolne regije mitohondrijske DNA kako bi se istražila
genetska struktura i povezanost populacija balkanske divokoze u cijelom rasponu
distribucije s ciljem razvoja strategije gospodarenja i zaštite. Osim toga, korišten je isti set
lokusa kako bi se utvrdila točnost postojećih povijesnih podataka o podrijetlu biokovske
populacije i razjasnilo koja je podvrsta bila prisutna prije reintrodukcije balkanske i alpske
divokoze na planinu Velebit s ciljem procjene i dokumentiranja genetskoga statusa izvornih
i translociranih populacija.
Za postizanje ciljeva istraživanja ukupno je genotipiziran 141 uzorak balkanske divokoze s
područja država Balkanskog poluotoka (osim Kosova) i 4 lubanje divojaraca (muzejski
primjerci). Za izradu filogenetske mreže, prikupljeno je 56 dodatnih uzoraka koji su pripadali
drugim podvrstama divokoza. Sekvenca kontrolne regije duga 376 parova baza uspješno
je očitana kod 44 jedinke.
STRUCTURE model predlaže 3 genetska klastera i svrstao je jedinke iz Srbije i Bugarske
u dva odvojena klastera, dok su jedinke iz ostalih zemalja pripadale istom klasteru.
Analizom sekvenci kontrolne regije mitohondrijske DNA otkriveno je 30 novih haplotipova s
privatnim haplotipovima u svim analiziranim populacijama i dva haplotipa dijeljena između
populacija, što može biti rezultat prošlih translokacija jedinki. STRUCTURE i GENELAND
analize pokazale su jasnu odvojenost reintroducirane populacije na Biokovu od divokoza
nastanjenih na Prenju i značajnu genetsku sličnost između biokovske populacije i populacija
uzorkovanih na Čvrsnici i Čabulji. GENELAND analiza prepoznala je balkansku divokozu s
Prenja kao zaseban genetski skup, različit od populacija koje obitavaju na Čvrsnici i Čabulji.
Ovi rezultati sugeriraju da su rijeka Neretva i državna cesta M17 geografske barijere za
rasprostranjenost podvrste. STRUCTURE i DAPC analize muzejskih uzoraka pokazale su
veću vjerojatnost da je na Velebitu prije reintrodukcije obitavala alpska podvrsta divokoze.
Rezultati ovog istraživanja su dali uvid u genetsku raznolikost populacija balkanske
divokoze i pokazali uzorak arhipelaga diljem rasprostranjenosti podvrste kao posljedica
fragmentacije staništa i populacija. Rezultati ovog istraživačkog rada mogu poslužiti kao
smjernice za izradu akcijskog plana gospodarenja populacijama balkanske divokoze u
Hrvatskoj, ali i u drugim zemljama gdje ova podvrsta obitava, te osigurati dizajn za buduću
zaštitu podvrste. |
| Abstract (english) | Chamois (Rupicapra spp.) are medium-sized ungulates that inhabit alpine pastures and
rocky areas on the main mountain massifs of both Europe and the Near East and,
exceptionally, the low elevations of river gorges, forested and coastal areas. The currently
accepted taxonomy of chamois, based on morphological, behavioral and molecular
evidence, recognizes two species: the Northern chamois (Rupicapra rupicapra) with seven
subspecies distributed on the Alps (R. r. rupicapra and R. r. cartusiana), the Balkans (R. r.
balcanica), the Tatras (R. r. tatrica) and the Carpathians (R. r. carpatica), in western Asia
(R. r. asiatica) and the Caucasus (R. r. caucasica). The Southern chamois (Rupicapra
pyrenaica) includes three geographically isolated subspecies on the Cantabrian Massif (R.
p. parva), the Pyrenees (R. p. pyrenaica) and the central Apennines (R. p. ornata). In the
early 1900s, chamois populations in the northern Dinaric Mountains in Croatia were
extirpated due to predation, natural events, and unsustainable hunting before their
taxonomic classification was assessed. Several decades after local extinction, the chamois
population was re-established through several reintroduction efforts between 1964 and
1978. This resulted in the current populations of Northern chamois in the northern Dinaric
Mountains being descended from successfully reintroduced individuals, captured in
mountainous areas in Bosnia and Herzegovina (Rupicapra r. balcanica) and Slovenia
(Rupicapra r. rupicapra), except for the Dinara massif, where the only Croatian
autochthonous population of Balkan chamois lives. Since different subspecies have been
involved in reintroduction in the past, a contact zone has formed in the northern Velebit
Mountains, where these subspecies hybridize.
The Balkan chamois is widespread on the Balkan Peninsula, along mountain massifs from
Croatia in the north to Greece in the south and Bulgaria in the east. The distribution of the
Balkan chamois is patchy and covers only parts of the massifs and mountain chains across
the countries that form its range. The subspecies has low rates of colonization and reduced
gene flow between isolated populations, which may result in genetic differentiation due to
the inbreeding effect and a loss of allelic variants as a consequence of genetic drift.
Reduced genetic diversity in these small and isolated populations might, in turn, cause
negative impacts on fitness, resulting in decreased effective population size and, eventually,
increase the probabilities of extinction. Other threats to the Balkan chamois survival are
considered to be poaching, introductions of other chamois subspecies (mostly Alpine
chamois), forest succession, road infrastructure, intensive livestock grazing, predation,
unsustainable hunting and natural events. Due to these threats, the Balkan chamois is
protected by Annexes II and IV of the European Union Habitats Directive 92/43/EEC (OJ L
206, 22.7.1992) and Appendix III of the Bern Convention (OJ L 38, 10.2.1982). The
conservation and management status within the different national legislations varies
between countries (members and non-members of the EU) and depends on the degree of
the local communities' interest in the conservation of the subspecies. Over the past two
decades, the genus Rupicapra has been the subject of numerous genetic studies and,
although Balkan chamois has been included in several of these phylogenetic studies that
used both nuclear and mitochondrial markers, it remains one of the less-studied
subspecies. Current knowledge on the genetic diversity and structure of the Balkan chamois
population is limited and restricted to regional-local studies. Multiplex set of microsatellite
loci and a partial mitochondrial control region were used to investigate the genetic structure
and connectivity of Balkan chamois throughout its distribution range to support the
development of management and conservation strategies. In addition, the same
microsatellite set was used to determine the accuracy of existing historical data on the origin
of the Biokovo population and to clarify which subspecies was present before reintroduction
of Balkan chamois to Velebit Mountain. This was done with the aim to assess and document
the genetic status of both the source and translocated populations.
To test Balkan chamois genetic diversity and population structure, DNA from bone, dried
skin and muscle tissue was extracted and successfully genotyped in 92 individuals of
Balkan chamois and the partial control region was sequenced in 44 individuals. Additional
20 samples from Biokovo and 29 samples from three areas which might have served as
source populations for reintroduction and possible recent recolonization (Prenj, Čvrsnica
and Čabulja mountains) were genotyped. To clarify which subspecies was present before
reintroduction to Velebit Mountain, four male chamois skulls originating from Velebit,
collected around 25 years before the population local extinction were genotyped.
The Bayesian analysis suggested that Balkan chamois are divided into 3 genetic clusters,
and assigned individuals from Serbia and Bulgaria to two separate clusters, while
individuals from the other countries belonged to the same cluster. Thirty new haplotypes
were obtained from partial mitochondrial DNA sequences, with private haplotypes in all
analysed populations and only two haplotypes shared among populations, indicating the
possibility of past translocations. Both STRUCTURE and GENELAND analyses showed a
clear separation of the reintroduced population on Biokovo from Prenj’ s chamois and
considerable genetic similarity between the Biokovo population and the Čvrsnica–Čabulja
population. This suggests that the current genetic composition of the Biokovo populations
does not derive exclusively from Prenj, as suggested by the available literature and personal
interviews, but also from Čvrsnica and Čabulja. GENELAND analysis recognized the Balkan
chamois from Prenj as a separate cluster, distinct from the populations of Čvrsnica and
Čabulja. This suggests that the Neretva River and the state M17 road are geographic
barriers for the species dispersal, as they form a genetic boundary. Concerning the
identification of the subspecies inhabiting Velebit Mountain before the reintroducitons,
according to assignment based on microsatellite loci, using both Bayesian clustering in
STRUCTURE (with q values between 0.55 and 0.73) and DAPC (with individual
membership probabilities of 0.99 and 1.00), Alpine subspecies showed an higher likelihood.
The subspecies’ genetic composition presented in this doctoral thesis provides the
necessary starting point for assessing the conservation status of Balkan chamois and allows
the development of strategies necessary for its sustainable management. The use of
molecular markers, in this case microsatellites and mitochondrial DNA, provided important
information on the genetic diversity and evolutionary history of Balkan chamois populations. |
