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.
Honey bee mite Varroa destructor and microsporidium Nosema ceranae are currently considered the most important threats to honey bees and beekeeping. It has been believed that both N. apis and N. ceranae invade exclusively epithelial cells of the honey bee ventriculus. However, some fi ndings suggest that these microsporidia may infect other tissues of honey bees. There are indications that these pathogens could be found in honey bee haemolymph, as the medium for its distribution to anatomically distant tissues. Knowing that V. destructor being an ectoparasitic mite feeds on the honey bee’s haemolymph, the aim of this study was to investigate if DNA of Nosema spp. microsporidia could be found in honey bee haemolymph and in V. destructor.
The study was conducted on bee haemolymph and V. destructor mites from 44 Apis mellifera colonies. From each hive five mite individuals and 10 μL of haemolymph (from 4-5 bees) were used as samples for DNA isolation and PCR detection of Nosema spp.
The DNA of N. ceranae was confi rmed in 61.36% of V. destructor mites and 68.18% of haemolymph samples. This is the first report of N. ceranae DNA in honey bee haemolymph and in V. destructor mites. The finding of DNA of N. ceranae in V. destructor could be interpreted as the result of mite feeding on N. ceranae infected bee haemolymph. However, for a full confi rmation of the vector role of V. destructor in spreading of nosemosis, further microscopy investigations are required for the detection of spores in both investigated matrices (haemolymph and V. destructor internal tissues).
Thelazia callipaeda is a parasitic nematode causing ocular infections in different mammalian species and humans, clinically manifested as lacrimation, conjunctivitis, keratitis, corneal opacity or corneal ulcer. In this paper, we reported six cases of autochthonous canine and feline thelaziosis on different localities in Serbia. Total of 285 parasites (85 males and 200 females) were collected from the eyes of infected animals (n=6) suffering from uni- or bilateral conjunctivitis, with the number of parasites ranging from 7 to 150 per animal. All parasites were morphologically identified as T. callipaeda, while molecular analyses of cytochrome oxidase 1 (cox1) gene revealed the presence of h1 haplotype, as the unique previously reported in other studies in Europe. Since T. callipaeda is a newly detected parasite in Serbia with the infective potential for humans, there is a necessity for animal owners education and cooperation among professional services in order to control this zoonosis.
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.
Vladimir Dimitrijevic, Jevrosima Stevanovic, Mila Savic, Branko Petrujkic, Predrag Simeunovic, Ivan Milosevic and Zoran Stanimirovic
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.
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.
Ivan Vujanac, Radiša Prodanović, Goran Korićanac, Jovan Bojkovski, Predrag Simeunović, Milija Palamarević, Sreten Nedić, Irena Celeska and Danijela Kirovski
This study aimed to evaluate glucose-induced insulin response in cows exposed to different temperature-humidity index. Twenty early lactating Holstein-Friesian cows were divided into 2 equal groups based on season, as summer (SU) and spring (SP). SP cows were not exposed to heat stress, while SU cows were exposed to moderate or severe heat stress. Milk production was recorded daily. Starting from day 30 of lactation, intravenous glucose tolerance test (IVGTT) was carried out three times at 30-day intervals. Blood samples were taken before (basal) and after glucose infusion, and glucose and insulin were measured at each sample point. The homeostatic model assessment (HOMA) index was calculated. Milk yield from days 30 to 40 and 64 to 90 of lactation were higher in SP cows than in SU cows. Basal glucose did not differ on days 30 and 60 of lactation, while basal insulin and HOMA were lower in SU compared to SP cows. On day 90 of lactation, SU cows had higher basal glucose, whereas basal insulin and HOMA did not differ. IVGTT results revealed that glucose tolerance was affected by heat stress such that SU cows had higher glucose clearance. Insulin responses to IVGTT did not differ on days 30 and 60 of lactation. Heat stress had a marked effect on insulin secretion on day 90 of lactation, illustrated by higher increments, peak concentrations and area under the curve for insulin in SU cows. Overall, season differences in glucose tolerance depend not only on heat stress and milk production but also on the stage of lactation.