Tihomir Florijančić

First evidence of hybridization between golden jackal (Canis aureus) and domestic dog (Canis familiaris) as revealed by genetic markers

Interspecific hybridization is relatively frequent in nature and numerous cases of hybridization between wild canids and domestic dogs have been recorded. However, hybrids between golden jackals (Canis aureus) and other canids have not been described before. In this study, we combined the use of biparental (15 autosomal microsatellites and three major histocompatibility complex (MHC) loci) and uniparental (mtDNA control region and a Y-linked Zfy intron) genetic markers to assess the admixed origin of three wild-living canids showing anomalous phenotypic traits. Results indicated that these canids were hybrids between golden jackals and domestic dogs. One of them was a backcross to jackal and another one was a backcross to dog, confirming that golden jackal–domestic dog hybrids are fertile. The uniparental markers showed that the direction of hybridization, namely females of the wild species hybridizing with male domestic dogs, was common to most cases of canid hybridization. A melanistic 3bp-deletion at the K locus (β-defensin CDB103 gene), that was absent in reference golden jackal samples, but was found in a backcross to jackal with anomalous black coat, suggested its introgression from dogs via hybridization. Moreover, we demonstrated that MHC sequences, although rarely used as markers of hybridization, can be also suitable for the identification of hybrids, as long as haplotypes are exclusive for the parental species.

The hygiene and quality of hare meat (Lepus europaeus Pallas) from Eastern Croatia

This research was conducted over two seasons (winter and spring) and comprised a total of 71 samples of hares shot in the eastern region of Croatia, and was aimed at examining the quality and hygiene of the meat. The degree of usefulness in relation to the total average weight of the shot hares was 66.54%. Chemical tests showed that the average amount of water in the meat of the hares amounted to 75.34%, protein 23.19%, fats 1.12% and ash 1.16%. Of the 71 samples examined, as many as 44 (61.97%) did not meet the requirements of the rules on microbiological food standards, of which in 19 samples (26.76%) this was due to the high level of aerobic mezophilic bacteria/g, 2 samples (2.82%) contained Staphylococcus aureus/g, and 37 samples (52.11%) were not satisfactory because they contained enterobacteria/g. In terms of heavy metals in kidney and meat samples, 17 (23.94%) did not meet the provisions of the rules on toxins, metals, metaloids and other harmful substances which may be found in food. Of these 15 kidney samples (88.24%) contained cadmium, and 2 samples (11.76%) mercury in amounts greater than the permitted concentration. Tests on liver samples from the hares (n=71) did not show organophosphorus pesticides, nor organochlorinated pesticides or polychlorinated biphenyls.

Trichinella pseudospiralis in pig from Croatia.

Trichinella spiralis and Trichinella britovi are species that are frequently found in domestic pigs and various sylvatic animals in Croatia. During routine trichinoscopy, non-encapsulated larvae were detected in the muscle tissue of a domestic pig. Artificial digestion revealed a larvae burden of 602 muscle larvae per gram of tissue. Tissue section analysis confirmed the presence of non-encapsulated larvae. Multiplex PCR identified the larvae as T. pseudospiralis. This observation is consistent with the reports of a local veterinary inspector who described the presence of non-encapsulated Trichinella in four individual cases over the last 2 years. This is the first report of T. pseudospiralis in Croatia and one of very few cases of T. pseudospiralis infection described in domestic pigs. The detection of non-encapsulated larvae stresses the need for implementation of artificial digestion instead of trichinoscopy for the detection and identification of Trichinella infections.

Heavy metal concentrations in the liver of two wild duck species: influence of species and gender

Abstract The risk of wild ducks exposure to heavy metals in the environment was assessed by analyzing 20 wild ducks shot in the hunting area at the fish farm in Donji Miholjac, Eastern Croatia. Liver samples obtained from 10 Mallards and 10 Common Pochards were examined for heavy metals (Cd, Pb, As, Hg) by using flame atomic absorption spectrophotometry (AAS). Significant difference in heavy metal content between analyzed species was confirmed only for Cd (P≤0.01), and no differences were found between genders within species. The results obtained suggest the importance of wild ducks as bioindicators of heavy metal pollution, especially Common Pochard for Cd, and Mallard for Pb accumulation.

THE EFFECTS OF ORGANIC SELENIUM AND MANNAN OLIGOSACCHARIDES ON THE PRODUCTIVITY AND HEALTH OF PHEASANT CHICKEN (PHASIANUS COLCHICUS)

The investigation included 37 pheasant chickens divided into three groups (control and two experimentals groups) which were all fed with a standard starter feed mixture (28% crude protein and 11.7 MJ ME/kg) for the first 28 days and then with a grower for pheasant chickens (24% of crude protein and 12.1 MJ ME/kg). Sel-Plex® was added to the first experimental group in a concentration of 0.2%. A biochemical investigation of the blood samples on the 70th day of the trial showed a significantly (P 0.05), when compared to the control.

Are the dinaric mountains a boundary between continental and mediterranean wild boar populations in Croatia

Wild boar is the most important large game species in Croatia, with populations subjected to hunting/management programs. There are indications from previous analyses of morphological variation and reproductive performance that population structure of wild boar in Croatia is strongly influenced by the Dinaric mountain range that separates Croatia in two biogeographic regions: Continental and Mediterranean. Of the two main wild boar haplogroups found in Europe, the most widespread is the E1 clade (present from Portugal to Russia), while the E2 clade is found within the Italian peninsula and on Sardinia. Outside Italy, the E2 clade has to date been found only in archaeological samples from the Mediterranean region of Croatia. The aims of this study were to investigate the hypotheses that (i) the Dinaric Mountains act as a natural boundary separating the Continental and Mediterranean wild boar subpopulations in Croatia; and (ii) the mtDNA E2 clade is present in Croatian wild boar. A total of 14 unlinked microsatellite loci were examined in 264 geo-referenced individuals, and the 486-bp fragment of the mtDNA control region sequence was examined in 69 individuals. The results suggest that the region of the Dinaric Mountains. represents a weak boundary between the Continental and Mediterranean subpopulations, and that the predominant factor causing the observed genetic structuring is isolation by distance. Based on these results, it can be proposed that the Croatian wild boar population in the mainland should be managed as a whole. Three variable sites and four haplotypes were obtained from the mtDNA sequences, all of which belonged to the major European clade (E1).

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.

STEROID HORMONES PROFILE DURING AN OVARIAN SYNCHRONIZATION PROCEDURE IN DIFFERENT AGE CATEGORIES OF RED DEER HINDS (Cervus elaphus L.)

The objective of the present study was to compare estradiol/progesterone ratios of different age categories of red deer hinds and use it as a predictor of estrus synchronization success and consequently conception rate. To accomplish this we used 38 red deer hinds to establish serum progesterone and estradiol levels in young (21 animals), mature (10 animals) and old (7 animals) hinds during the estrus synchronization procedure (transvaginal/cervical AI). The following estrus synchronization was used: at the start of the experiment each hind received a controlled intravaginal drug-releasing device (CIDR, Pharmacia&Upjohn, New Zealand) containing 0.3 g of progesterone. The device was removed on day 11, simultaneously with an application of 250 IU of Pregnant Mare Serum Gonadotropin (PMSG, Folligon® Intervet International, Boxmeer, Holland). Transvaginal/cervical AI (artificial insemination) was performed 48 hours after CIDR withdrawal (day 13). Blood samples were obtained from the jugular vein using a Venoject® vacutainer without an anticoagulant for hormonal tests on the same experimental day (0, 11th and 13th day). A statistically (p<0.01) higher progesterone level was found in young hinds on the 11th day after controlled intravaginal drug-releasing device insertion. A significantly higher (p<0.01) estrogen level was observed in the young in regard to mature and old hinds on the expected day of estrus (13th day). Estradiol/progesterone ratios showed a statistically significant difference (p<0.01) on insemination day (13th day) between old and young hinds (98.67 : 46.59) and between old and mature hinds (98.67 : 51.79). Out of a total of 38 hinds only 9 had their offspring, 6 of the young and 3 of the mature hinds.

Preliminary serological and molecular investigation of selected viral pathogens in Croatian cervid species

A total of 131 blood samples and 175 spleen samples were collected from three cervid species: roe deer (Capreolus capreolus), red deer (Cervus elaphus) and fallow deer (Dama dama) inhabiting the continental part of Croatia. Serum samples were tested for antibodies against bovine herpesvirus 1, parainfl uenza-3 virus, bluetongue virus, bovine respiratory syncytial virus, hepatitis E virus, bovine viral diarrhoea virus and enzootic bovine leukosis virus. The tested sera were negative for bovine viral diarrhoea virus, enzootic bovine leukosis virus, bluetongue virus, bovine respiratory syncytial virus and hepatitis E virus antybodies. The antibody prevalence in roe deer and red deer samples was 21.11% for bovine herpesvirus 1 and 75.55% for parainfl uenza-3 virus. Sera from bovine herpesvirus 1 positive animals were subsequenty tested with comparative virus neutralization test and bovine herpesvirus 1 neutralising antibodies were found in 18 (out of 19) sera. In fallow deer, no antibodies against any of the viral pathogens were detected. All spleen samples tested for bovine viral diarrhoea virus and enzootic bovine leukosis virus came back negative, except for one red deer spleen sample found to be weakly diarrhoea virus-positive. Our fi ndings provide the fi rst information on the exposure of Croatia-inhabiting cervid species to viral pathogens, and could serve as valuable baseline data for future investigations regarding deer exposure to various pathogens and the distribution of diseases shared between wildlife and livestock. As of now, the epidemiology of these viruses in the Croatian cervid population has been only poorly understood, so that further research is recommended.

Habitat conditions for wildlife management in the Dinaric regions of Croatia and Montenegro

Hunting is an important socio-economic activity, particularly in rural areas. Characteristics and quality of habitat are important in the sustainable management of game animals and wildlife conservation. Three hunting grounds were analysed and their habitat composition was compared with regard to abundance of main game animal species. Hunting ground VIII/2 “Bjelolasica” is located in Gorski kotar region; XVII/6 “Maglaj-Cista” in the inland of middle Dalmatia; and hunting ground “Dragišnica” is located in northern part of Montenegro.