Predrag Simeunovic

Morphological, biochemical and hematological characterization of endangered Balkan donkey breed.

Abstract The aim of the study was to establish morphometric, biochemical and hematological values for the endangered Balkan donkey breed (Serbia) and to explore the possible age dependence of the parameters tested. Inter-breed similarity of morphometric parameters was assessed by comparing the data obtained for the Balkan donkey with morphometric measurements of several previously characterized domestic donkey breeds. The study population included 74 donkeys, divided in two age groups (group A ≤ 3 years; group B > 3 years). In total, 18 morphometric, 13 hematological and 14 biochemical parameters were assessed. Significant morphometric differences (p<0.05) in body length, head length, chest circumference and body weight were found between the two age groups. Significant differences in morphological parameters were revealed among the Balkan donkey and other donkey breeds (Catalonian, Croatian and Albanian), but results of cluster analysis demonstrated the smallest distance between the Balkan donkey and Albanian donkeys. The results of morphometric analyses showed consistency of the obtained values within the breed, and diversity as compared to other donkey breeds, and, thus, could be taken as referent for the Balkan donkey. Hematological and biochemical profiles obtained for the Balkan donkey were consistent with previous reports and within the recommended reference ranges. White blood cell, mid cell and granulocyte counts, showed significantly higher (p<0.05) values in donkeys under 3 years of age, while the only biochemical parameter affected by age was alkaline phosphatase. The information gained through characterization of the Balkan donkey breed provides a basis for conservation and development of the breed standard.

Nosema ceranae and queen age influence the reproduction and productivity of the honey bee colony

Summary The strength of the honey bee colony and the quality concerning brood production, number of worker bees and honey yield depend substantially upon the characteristics of the queen. Reproductive and productive capacities of the colony, as two important physiological qualities can depend a great deal on the age of the queen. Over the years, the laying ratio of queens could decrease depending on the age; therefore colonies headed by one-year-old queens have proven to be more efficient considering colony population and honey production than ones with older queens. Nosema ceranae infection is a possible reason for the decrease in reproductive capacities and the reduction in the honey production of infected colonies. With the help of numerous pheromones, the queen can preserve colony homeostasis, however exhausted by N. ceranae infection she might compromise the renewal of worker population. The aim of this research is to investigate the influence of N. ceranae on reproduction and productivity of honey bee colonies headed by queens of different age. Experiment was conducted on 27 honey bee colonies equalized with regard to brood, food source, number of worker bees and Nosema infection. In the three year investigation period (2009–2012), Nosema detection, identification and spore load per each colony, as well as parameters of colony reproduction (such as uncapped and capped brood area, queen egg-laying rate and colony strength) and productivity (the total extracted honey and the winter honey stores) were recorded. Concerning the influence of the age of the queens on followed parameters we compared one, two and three-years old queens and found significantly higher reproductive and productive values in favour of younger ones. Significant differences regarding the effects of N. ceranae infection on productive and reproductive characteristics could be explained with a higher capacity of one year old queens to compensate the effects of Nosema infection; a capacity which gradually declines in the second and the third year.

Variability of the honey bee mite Varroa destructor in Serbia, based on mtDNA analysis

Only two mitochondrial haplotypes (Korea and Japan) of Varroa destructor, the ectoparasitic honey bee mite, are known to be capable of infesting and successfully reproducing in Apis mellifera colonies worldwide. Varroadestructor (then called Varroa jacobsoni) was observed in Serbia for the first time in 1976. In order to obtain insight into the genetic variability of the mites parasitizing A. mellifera we analyzed 45 adult female mites sampled from nine localities dispersed throughout Serbia. Four fragments within cox1, atp6, cox3 and cytb mtDNA genes were sequenced. The Korea haplotype of V. destructor was found to be present at all localities, but also two new haplotypes (Serbia 1 and Peshter 1) were revealed, based on cox1 and cytb sequence variability. The simultaneous occurrence of Korea and Serbia 1 haplotypes was observed at five localities, whereas Peshter 1 haplotype was identifed at only one place.

Retrospective Analysis of the Bluetongue Outbreak in Serbia

Abstract Bluetongue, a vector-born disease caused by the Bluetongue virus (BTV) and transmitted by Culicoides biting midges, is considered to be one of the most important diseases of domestic ruminants. The first outbreak of bluetongue in Serbia was reported in 2001, when BTV serotype 9 was identified in sampled materials. In 2014, outbreak of BTV-4 in Serbia caused considerable economic losses affecting sheep, cattle and goats. During this outbreak, BTV-4 was recorded in 644 outbreaks within 49 municipalities, part of 17 administrative regions. From the total number of sheep kept in areas affected by bluetongue (n=1 748 110), 2 083 cases (0.2%) were proven to be BTV-4 infected. Total of 206 infected cattle and 24 infected goats were reported during this investigation period, which represents 0.06% and 0.03% of the total number of cattle and goats kept in affected areas, respectively. The highest incidence of infected sheep, cattle and goats was recorded on the territory covered by veterinary institute of Nis. Recorded lethality in cattle, sheep and goats was 18.45% (n=38), 48.10% (n=1002) and 54.17% (n=13), respectively. The peak of the outbreak was in September and October when 94.43% of the confirmed positive cases, regardless of the species, was recorded. Monitoring of bluetongue disease in Serbia relies on active surveillance programmes aimed at: (i) identification and tracing of susceptible and potentially infected animals and (ii) detection, distribution and prevalence of insect vectors. Vaccination of sheep is planned to be implemented as a control measure against bluetongue in Serbia.

A survey of deformed wing virus and acute bee paralysis virus in honey bee colonies from serbia using real-time RT-PCR

Abstract In this study 55 honey bee colonies from different Serbian regions were monitored for the presence of Deformed Wing Virus (DWV) and Acute Bee Paralysis Virus (ABPV) using TaqMan-based real-time RT-PCR assay. The results revealed the presence of DWV in each sampling location, and ABPV in 10 out of 11 apiaries. High frequency of DWV (76.4%) and ABPV (61.8%) positive samples in asymptomatic colonies can be the consequence of inefficient and postponed Varroa treatment concerning the role of this mite in the transmission and activation of honey bee viruses. The real-time RTPCR technique described in this paper is proved to be the most reliable method for this kind of investigation.

Validation of 10 Microsatellite Loci for Their use in Parentage Verification and Individual Identification in the Yugoslavian Shepherd Dog Sharplanina / Walidacja 10 Loci Mikrosatelitarnych Do Kontroli Pochodzenia Oraz Identyfikacji Osobniczej Jugosłowiańskiego Psa Pasterskiego Z Szar Płaniny

Abstract The aim of the study was to assess a commercially available microsatellite panel for use in paternity and identification analyses in the Yugoslavian Shepherd Dog Sharplanina, an ancient livestock guarding breed. Allele frequencies for 10 microsatellite loci (PEZ01, FHC2054, FHC2010, PEZ05, PEZ20, PEZ12, PEZ03, PEZ06, PEZ08 and FHC2079) were determined in 103 unrelated Yugoslavian Shepherd Dog individuals. The loci revealed varied levels of polymorphism (five to 12 alleles), with an average of 7.83 per locus. Average values of observed heterozygosity and polymorphic information content (PIC) were 0.64 and 0.66, respectively. Nine out of 10 microsatellite markers were highly informative with PIC values higher than 0.5. The obtained value of combined power of exclusion (0.9989) confirms usefulness of this panel of microsatellites for parentage verification, while the value of combined power of discrimination of 0.9999 clearly shows that the panel can conclusively identify individual dogs. In conclusion, the results obtained suggest that the selected set of commercially available microsatellite markers may be used as a routine tool for parentage verification and individual identification in the Yugoslavian Shepherd Dog. Although analysis of genetic variability of the Yugoslavian Shepherd Dog was not the primary focus of the study, the established values of major diversity indices disclose a highly variable gene pool in the breed. Streszczenie Celem eksperymentu była ocena przydatności komercyjnego panelu markerów mikrosatelitarnych do analizy pochodzenia i identyfikacji osobniczej owczarka jugosłowiańskiego Sarplaninac, starodawnej rasy psów pasterskich. Frekwencje alleli dla 10 loci mikrosatelitarnych (PEZ01, FHC2054, FHC2010, PEZ05, PEZ20, PEZ12, PEZ03, PEZ06, PEZ08 i FHC2079) określono u 103 niespokrewnionych osobników rasy Sarplaninac. W loci ujawniono zróżnicowany poziom polimorfizmu (5 do 12 alleli), wynoszący średnio 7,83 na locus. Średnie wartości heterozygotyczności obserwowanej i współczynnika polimorfizmu (PIC) wyniosły, odpowiednio 0,64 i 0,66. Dziewięć z 10 markerów mikrosatelitarnych było wysoko informatywnych, przy wartościach PIC przekraczających 0,5. Uzyskana wartość skumulowanej siły wykluczenia (0,9989), potwierdza przydatność badanego panelu markerów mikrosatelitarnych do kontroli pochodzenia, natomiast wartość skumulowanej siły dyskryminacji, wynoszącej 0,9999 wyraźnie wskazuje, że panel ten może jednoznacznie identyfikować poszczególne osobniki. Uzyskane wyniki wskazują, że wybrany zestaw komercyjnych markerów mikrosatelitarnych może służyć jako rutynowe narzędzie do kontroli pochodzenia oraz identyfikacji osobniczej jugosłowiańskiego psa pasterskiego. Choć analiza zmienności genetycznej owczarka jugosłowiańskiego nie była głównym celem badań, uzyskane wartości głównych indeksów różnorodności wskazują na występowanie wysoko zróżnicowanej puli genowej w tej rasie.

The effect of Agaricus brasiliensis extract supplementation on honey bee colonies

This study was done to discover any beneficial effect of a medicinal mushroom Agaricus brasiliensis extract on the honey bee. Firstly, a laboratory experiment was conducted on 640 bees reared in 32 single-use plastic rearing cups. A. brasiliensis extract proved safe in all doses tested (50, 100 and 150 mg/kg/day) irrespective of feeding mode (sugar syrup or candy). Secondly, a three-year field experiment was conducted on 26 colonies treated with a single dose of A. brasiliensis extract (100 mg/kg/day) added to syrup. Each year the colonies were treated once in autumn and twice in spring. The treatments significantly increased colony strength parameters: brood rearing improvement and adult population growth were noticed more often than the increase in honey production and pollen reserves. These positive effects were mainly observed in April. In conclusion, A. brasiliensis extract is safe for the bees and helps maintaining strong colonies, especially in spring.

A molecular genetic approach to roebuck individual identification in the case of poaching in Serbia

Application of the molecular genetic methods in forensic cases dealing with wild animals has significantly increased recently. These techniques are practically used in order to help solving four key problems : determination of kind of the wild animal, geographic origin, kinship ties and individual identification. In this work the first case of introducing the examination of polimorphism of microsatelite genetic markers within forensic analysis in the cases of poaching in Serbia is presented. The objectives of this forensic analysis was to determine if the meat confiscated during house search of the suspect comes from roebuck origin (Capreolus capreolus), which remains had been found by a game warden in the field during closed season, where the suspect denied the offense, claiming that the meat comes from other roebuck that had been shot during the previous hunting season. DNK was isolated from the skin and fur samples taken from the roebuck corpse found in the woods, as well as from the frozen meat found in the suspect’s house. Both amplification and polimorphism examination of the eight microsatelite markers (ROE01, NVHRT21, NVHRT24, NVHRT48, NVHRT73, RT7 AND RT27) were carried out. In all the examined samples, the same pattern of variability of the tested microsatelites was determined, that is it was proved that DNK profiles of the samples taken from roebuck corpse were identical to DNK profile of the meat sample found in the suspect’s house. This result clearly indicates that all the examined biological samples originate from the same animal, and consequently represents forensically valid evidence in the case of roebuck poaching. [Projekat Ministarstva nauke Republike Srbije, br. III46002]