Magda Sindičić

A multidimensional approach to managing the European brown bear in Croatia

Abstract Because of its biological characteristics, its important place in the minds of humans, and the considerable international interest for its conservation, management of the brown bear (Ursus arctos) in Europe is challenging. The Brown Bear Management Plan for Croatia (BMPC) was approved in 2004 and addressed interests such as ecology, aesthetics, and economics, as well as concern for the safety of people and property. It attempts to ensure conditions for the long-term survival of the brown bear, a species listed as endangered by some international regulations but as a game species (subject to regulated hunting) in others, including Croatia. Careful evaluation of actions affecting population size represents the most critical part of this plan. Those actions should sustain long-term viability of the bear population while maintaining densities at a level that minimize human–bear conflict. To achieve this goal, a series of actions and measures have to be regulated that are related to (1) bear habitat, (2) human activities in the habitat (e.g., highway construction, feeding of bears by humans), (3) prevention of problematic bear occurrences, and (4) the scientific monitoring of population changes. Although the plans development and implementation is the responsibility of bear management experts, various interests groups were considered. In large carnivore management, and especially in bear management, there are no final and universal solutions. Changes in the number of bears, areas of their presence, or behavior require new decisions. This plan offers guidelines for the decision-making process, and, because it includes a revision process, can be adapted to address new circumstances that arise. Citizens interested in conservation, not only in Croatia but also in neighboring countries, expect Croatia to work toward maintaining the long-term existence of as many bears as possible in appropriate habitats, with as few negative effects as possible. This plan is an important step in fulfilling those expectations.

Serologic evidence of Leptospira spp. serovars in brown bears (Ursus arctos) from Croatia.

Serum samples from 52 free-ranging brown bears (Ursus arctos) collected in Croatia over a period of 10 yr (1998–2007) were tested by microscopic agglutination test for specific antibodies (Ab) to 12 Leptospira spp. pathogenic serovars. At titers ranging from 1:100 to 1:2,000, 19 samples (36.5%) were Ab-positive to at least one serovar. Antibodies for 10 Leptospira spp. serovars were detected: Icterohaemorrhagiae, Australis, Sejroe, Canicola, Poi, Hardjo, Ballum, Saxkoebing, Pomona, and Grippotyphosa. In comparison to previous reports, the prevalence of Ab to serovar Icterohaemorrhagiae (52.6%) was significantly higher. Other common serovars were Australis (47.4%) and Sejroe (42.1%). High Ab titers for serovars Canicola (1:500) and Grippotyphosa (1:1,000) were detected for the first time in free-ranging bears from Croatia. A significant correlation between the age of the bears and detection of Ab to Leptospira spp. serovars suggested the presence of pathogenic agents in the natural habitats, whereas increasing trends of Ab prevalence for specific serovars (Icterohaemorrhagiae, Australis, and Sejroe) confirmed cohabitation of bears with rats and other small terrestrial mammals on garbage dumps and at bear feeding stations. To prevent cohabitation of bears and rodents, improvements in Croatian waste treatment, big game management, and rodent control programs are strongly recommended, especially in Lika and Gorski Kotar, regions that have high-quality natural habitats for brown bears in Croatia.

Repetitive sequences in Eurasian lynx (Lynx lynx L.) mitochondrial DNA control region

Mitochondrial DNA (mtDNA) control region (CR) of numerous species is known to include up to five different repetitive sequences (RS1–RS5) that are found at various locations, involving motifs of different length and extensive length heteroplasmy. Two repetitive sequences (RS2 and RS3) on opposite sides of mtDNA central conserved region have been described in domestic cat (Felis catus) and some other felid species. However, the presence of repetitive sequence RS3 has not been detected in Eurasian lynx (Lynx lynx) yet. We analyzed mtDNA CR of 35 Eurasian lynx (L. lynx L.) samples to characterize repetitive sequences and to compare them with those found in other felid species. We confirmed the presence of 80 base pairs (bp) repetitive sequence (RS2) at the 5′ end of the Eurasian lynx mtDNA CR L strand and for the first time we described RS3 repetitive sequence at its 3′ end, consisting of an array of tandem repeats five to ten bp long. We found that felid species share similar RS3 repetitive pattern and fundamental repeat motif TACAC.

When things go wrong: Cysticercus longicollis in an adult wild red fox (Vulpes vulpes)

First case of Cysticercus longicollis, larval stage of Taenia crassiceps, was diagnosed in a wild adult male red fox (Vulpes vulpes). The fox was killed by dogs at Nature Park Medvednica and presented to the University of Zagreb Faculty of Veterinary Medicine with history of being unable to run away and having skin lesions on legs that resembled to those of mange. Necropsy revealed whitish fluctuant mass full of cysticercus-like structures, surrounded by fibrous capsule and placed between the leg muscles, and numerous of spherical cysts in the subcutis and in the peritoneal cavity. Cysticerci were identified as C. longicollis based on their size, number and size of the rostellar hooks, mode of proliferation and DNA analysis. To our knowledge, this is the first case of T. crassiceps cysticercosis in a wild carnivore.

PCR-based Y chromosome marker for discriminating between golden jackal (Canis aureus) and domestic dog (Canis lupus familiaris) paternal ancestry

AbstractNumerous reports of hybridization between wild Canis species and domestic dog suggest that hybridization between golden jackal and dog is likely. Here we present an assay of a Y chromosome marker for discriminating between golden jackal (Canis aureus) and dog (Canis lupus familiaris) paternal ancestry. Taking advantage of an insertion found in a dog Zfy intron haplotype but not in a golden jackal haplotype, we developed a three-primer PCR system in which species are differentiated based on the number of amplicons. Two amplicons are produced from domestic dog DNA template, while one amplicon is produced from golden jackal DNA template. Both amplicons can be analyzed in a single agarose gel electrophoresis run, while the longer amplicon also serves as an internal control. The method was validated using 27 golden jackal and 25 dog samples; all samples showed the expected amplicons. This simple and cost-effective method should prove useful for determining the direction of golden jackal–dog hybridization.

Molecular identification of the rumen flukes Paramphistomum leydeni and Paramphistomum cervi in a concurrent infection of the red deer Cervus elaphus.

Paramphistomosis, caused by paramphistomid flukes, is a gastrointestinal parasitic disease of domestic and wild ruminants. Originally thought to be limited to the tropics and subtropics, the disease has recently been reported in temperate regions. Here we describe the concurrent infection of a red deer doe (Cervus elaphus) with Paramphistomum leydeni and Paramphistomum cervi. This is the first report of P. leydeni in Croatia. Flukes were identified on the basis of morphological keys (tegumental papillae) and sequencing of the internal transcribed spacer region 2 in ribosomal DNA. Our results confirm that the absence of tegumental papillae allows P. cervi to be differentiated morphologically from other paramphistomid species in Europe based on incident light stereomicroscopy. Nevertheless the limitations of morphological identification and taxonomic issues suggest that previous findings on paramphistomid infection should be interpreted carefully. The possible worldwide distribution of these pathogens means that paramphistomosis may be more common and its economic impact greater than previously thought.

Targeted resequencing of coding DNA sequences for SNP discovery in nonmodel species

Targeted capture coupled with high‐throughput sequencing can be used to gain information about nuclear sequence variation at hundreds to thousands of loci. Divergent reference capture makes use of molecular data of one species to enrich target loci in other (related) species. This is particularly valuable for nonmodel organisms, for which often no a priori knowledge exists regarding these loci. Here, we have used targeted capture to obtain data for 809 nuclear coding DNA sequences (CDS) in a nonmodel organism, the Eurasian lynx Lynx lynx, using baits designed with the help of the published genome of a related model organism (the domestic cat Felis catus). Using this approach, we were able to survey intraspecific variation at hundreds of nuclear loci in L. lynx across the species’ European range. A large set of biallelic candidate SNPs was then evaluated using a high‐throughput SNP genotyping platform (Fluidigm), which we then reduced to a final 96 SNP‐panel based on assay performance and reliability; validation was carried out with 100 additional Eurasian lynx samples not included in the SNP discovery phase. The 96 SNP‐panel developed from CDS performed very successfully in the identification of individuals and in population genetic structure inference (including the assignment of individuals to their source population). In keeping with recent studies, our results show that genic SNPs can be valuable for genetic monitoring of wildlife species.

First identification of Echinococcus multilocularis in golden jackals in Croatia

Alveolar echinococcosis, caused by the tapeworm Echinococcus multilocularis, is one of the world’s most dangerous zoonosis and an emerging disease with growing incidence in humans. The disease has been reported in new areas and host species in the last two decades, and the primary hosts of the parasite — red fox, golden jackal and grey wolf — are expanding their distribution in Europe. Here we report the morphological and molecular identification of Echinococcus multilocularis tapeworms in one of 29 carcasses of adult golden jackals in Croatia, where the only previous report of the parasite was in red foxes in 2016. These results suggest that alveolar echinococcosis should be treated as an emerging disease in Croatia.