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The “beautiful viviparous fly”, Wohlfahrtia magnifica, may have a magnificent appearance due to its striking morphology; however, it is a potentially deadly agent of obligate traumatic myiasis in humans and animals, with a serious impact on welfare and economics. The fly is found across the Palearctic realm, including the Western Balkan region, with reports from former Yugoslavian countries from the first half of the 20th century. In this paper, a recent case of wohlfahrtiosis recorded in Northern Serbia is evidenced using morphological and molecular techniques. Larvae were collected from two adult sheep with severe hoof myiasis and two young sheep with genital and interdigital myiasis. Morphological identification was performed for adults bred from the infested vulva and third-stage larvae (L3) collected from the hoof wounds, supported with barcoding sequences of the COI gene obtained from larval pairs from the hoof wounds of older and the genitalia of younger sheep. W. magnifica was identified according to the appearance of male fly terminalia and the morphology of L3, which was confirmed after the comparison of representative sequences of the COI gene (deposited in GenBank™ under accession numbers MT027108–MT027114) to those available in GenBank™. This finding represents the first reported case of wohlfahrtiosis in the Western Balkans in 80 years, highlighting the need to re-inform relevant stakeholders to achieve adequate disease control.

Purpose: Data on gastrointestinal infections in horses in Southeastern Europe are limited; thus, this study aimed to add to the existing knowledge on this topic by reporting on the prevalence of intestinal parasites of horses in the territory of the Republic of Serbia. Methods: In the period from April 2017 to December 2018, parasitological examination of 548 samples from horses of different breed, age and sex from four regions and 18 districts of the country was performed. Coprological diagnostic was done by using qualitative methods without concentration and qualitative methods with concentration of parasitic elements. Quantification of the obtained results was performed using semi-qualitative faecal egg count. Results: Four helminthoses were detected in the examined samples: P. equorum (8.57%), O. equi (3.65%), strongylid eggs (71.17%) and Anoplocephala spp. (0.91%). The total prevalence of helminthoses was 77.19%. Monoinfections were significantly more present 70.07% compared to coinfections (7.12%). The highest prevalence of helminthoses was detected in free-ranging horses (93.10%—27/29), in autumn 86.67% (117/135) and winter 79.71% (165/207), in Šumadija and West Serbia region (100%), and in the youngest category (100%). Significant difference (p < 0.001) was detected in the prevalence of monoinfections by strongylids and O. equi and also coinfections by strongylid/P. equorum between horses of different age categories. Conclusion: Obtained results are of great contribution to clinical parasitology and pathology, especially from the aspect of animal health, welfare and preservation of horse population.

Background. Urinary capillariosis in dogs is caused by Capillaria plica (syn. Pearsonema plica), a ubiquitous parasitic nematode resembling a string which belongs to the family Capillariidae. It parasitizes the feline, canine and musteline urinary bladder, and has been found in ureters and renal pelvises as well. C. plica has an indirect life cycle, with earthworms (Lumbricina) as intermediate hosts and domestic and wild animals (dog, cat, fox and wolf) as primary hosts. Infection of primary hosts occurs via ingestion of earthworms that contain infective first stadium (L1) larvae. An alternative path of infection for primary hosts is assumed to be ingestion of soil contaminated by infectious larvae derived from decomposed earthworms. Infection is mostly asymptomatic, but the clinical picture presents with pollakiuria, dysuria, haematuria, polydipsia, incontinence and/or fever. Scope and Approach. The aim of this review is to highlight the importance of urinary capillariosis in dogs. Since the health care of wild and domestic carnivores is extremely important, this review provides information about the morphology, biology and epizootiology of the C. plica nematode. Due to the importance of this disease for clinicians and increased disease prevalence during the last decade in many countries, this review presents the latest information on the pathogenesis, clinical signs, diagnosis, treatment and prevention of this infection. Key Findings and Conclusions. Capillariosis is usually accidentally diagnosed due to the nonspecific clinical signs and there is no treatment of choice. Practitioners should consider latent urinary capillariosis infection as a possible cause while examining for urinary tract diseases., Uvod. Urinarnu kapilariozu pasa uzrokuje Capillaria plica (syn. Pearsonema plica), ubikvitarna nematoda končastog oblika iz familije Capillariidae. Uzročnik parazitira u mokraćnoj bešici kanida, felida i mustelida, a može da se ustanovi u ureterima i bubrežnoj karlici. Capillaria plica ima indirektan životni ciklus koji uključuje kišne gliste (Lumbricina) kao prelazne domaćine i domaće i divlje životinje (pas, mačka, lisica, vuk) kao prave domaćine. Infekcija pravog domaćina nastaje unošenjem kišnih glista koje sadrže infektivnu larvu prvog stadijuma (L1). Pretpostavlja se da alternativni put infekcije za životinje može da bude ingestija zemljišta kontaminiranog infektivnim larvama poreklom od kišnih glista koje su podlegle procesu dekompozicije. Infekcija je najčešće asimptomatska, a u slučaju ispoljavanja kliničke slike simptomi su polakiurija, disurija, hematurija, polidipsija, inkontinencija urina i groznica Cilj i pristup. Cilj rada je da pruži informacije o morfologiji, biologiji i epizootiologiji nematode Capilaria plica kao i da ukaže na mogućnost postojanja infekcije kod lovačkih i nevlasničkih pasa. Zbog značaja oboljenja za kliničare i povećanja prevalencije tokom poslednje decenije u zemljama u okruženju, u radu su iznete najnovije informacije o patogenezi, kliničkim simptomima, dijagnostici, lečenju i prevenciji ove nematodoze. Ključni nalazi i zaključak. Dijagnoza kapilarioze se najčešće postavlja slučajnim nalazom parazita zbog nespecifičnе kliničke slike. U slučaju sumnje na oboljenja urinarnog trakta, treba diferencijalno dijagnostički isključiti latentnu infekciju ovom nematodom.

In the last decade, as the result of climate changes, there have been considerable changes in the parasitofauna of domestic and wild carnivores. Th e prevalence of the existing parasitic species has varied signifi cantly, showing an increasing tendency, and some parasitic species not present before in this epizootiologic territory have been diagnosed as well. It is thought that the reason for such an epizootiological situation is increased presence of owners with their pet animals in the regions endemic for particular zoonotic helminthoses during summer holidays and touristic visits. Th is tendency has become especially conspicuous in the last several years characterized by warm winters and very hot summers due to global warming eff ects, with abundant atmospheric precipitation. Oral vaccination of foxes against rabies, regulated in Serbia by appropriate laws since 2010, has led to an increased number of foxes and rise of prevalence of the parasitic diseases for which foxes represent the infection source/reservoir. Continued urbanization of Serbian cities, with the extension of urban belts into the suburbia and recreational (“weekend”) settlements, lead to a closer contact of street dogs and owned dogs with foxes, which results in a signifi cant change in the parasitic fauna of dogs. It is an additional factor, which in the chain fox - street dog - owned dog - human increases the risk and tendency for the occurrence of human infections with zoonotic endoparasites of wild and domestic carnivores. In order to reliably predict the degree of spread of particular zoonotic helminthoses in particular regions in Serbia, for which wild carnivores represent the infection source, it is necessary to institute continued monitoring of the parasitic fauna in this type of wild animals.

Control of the gastrointestinal parasites in different systems of ruminant breeding is based on vaccination, chemotherapy, improved herd management and use of genetic potentials of host animals. Strategy of the helminths control based on frequent anthelmintics usage is dominant among the world, although it is considered unsustainable due to the appearance of increased number and species of parasites that are resistance to drugs. Development of resistance on all three groups of broad-spectrum anthelmintics (nicotinic anthelmintics, benzimidazoles and macrolides lactones) as well as increased care for consumers health caused by appearance of used drugs residues in food, additionally complicate overall nematode control. With the aim to decrease appearance of anthelmintics resistance of gastrointestinal nematodes of small ruminants and in order to organize process of sustainable integrated parasite management, principle of targeted selective treatment is globally accepted. Implementing of this strategy has only recently become feasible, with development and practical use of systems that serve for clinical assessment of anemia in small ruminants which suffer from hemonchosis. Besides that, short term changes in body weight and body condition scoring may be indicators of diseases caused by endoparasites, as it can provide rapid identification of aminals that will probably have benefits from therapy. Obtained results of quantitative coprological diagnostic tests and results for anemia assessment are criteria that provide differential diagnosis between healthy and resilient animals and easier diseases diagnostic. Since resilient animals play important role in pasture contamination, the significance of their detection in understandable.

Kontrola gastrointestinalnih parazita u različitim sistemima uzgoja preživara zasnivana je na vakcinaciji, hemoterapiji, poboljšanju kvaliteta menadžmenta stada i korišćenju genetskih potencijala domaćina. Širom sveta dominira strategija kontrole helminata zasnovana na čestoj upotrebi antihelmintika, koja se smatra neodrživom, s obzirom na pojavu sve većeg broja i vrsta parazita koji su otporni na lekove. Razvoj rezistencije na sve tri grupe antihelmintika širokog spektra (nikotinski antihelmintici, benzimidazoli i makrolidni laktoni) i povećana briga za zdravlje potrošača uslovljene pojavom rezidua primijenjenih lijekova u hrani, dodatno komplikuju kontrolu ovih nematoda. U cilju smanjenja brzine pojave rezistencije na antihelmintike kod gastrointestinalnih nematoda malih preživara i organizacije procesa odživog integrisanog upravljanja parazitima, globalno je prihvaćen princip ciljanog selektivnog tretmana. Sprovođenje ove strategije je tek odnedavno postalo izvodljivo, sa razvojem i praktičnom primenom sistema koji služi za kliničku procjenu anemije kod malih preživara obolelih od hemonhoze. Pored toga, kratkotrajne promene telesne težine i određivanje telesne kondicije mogu biti pokazatelji endoparazitoza, što omogućava brzu identifikaciju životinja koje će verovatno imati koristi od tretmana. Dobijeni rezultati kvantitativne koprološke dijagnostike i rezultati procene anemije su kriterijumi koji omogućavaju diferencijalnu dijagnostiku između zdravih i rezilijentnih životinja i lakšu dijagnostiku rezistencije parazita. S obzirom na to da rezilijentne životinje igraju važnu ulogu u kontaminaciji pašnjaka, razumljiv je kliničko-parazitološki značaj njihovog pravovremenog otkrivanja., Control of the gastrointestinal parasites in different systems of ruminant breeding is based on vaccination, chemotherapy, improved herd management and use of genetic potentials of host animals. Strategy of the helminths control based on frequent anthelmintics usage is dominant among the world, although it is considered unsustainable due to the appearance of increased number and species of parasites that are resistance to drugs. Development of resistance on all three groups of broad-spectrum anthelmintics (nicotinic anthelmintics, benzimidazoles and macrolides lactones) as well as increased care for consumers health caused by appearance of used drugs residues in food, additionally complicate overall nematode control. With the aim to decrease appearance of anthelmintics resistance of gastrointestinal nematodes of small ruminants and in order to organize process of sustainable integrated parasite management, principle of targeted selective treatment is globally accepted. Implementing of this strategy has only recently become feasible, with development and practical use of systems that serve for clinical assessment of anemia in small ruminants which suffer from hemonchosis. Besides that, short term changes in body weight and body condition scoring may be indicators of diseases caused by endoparasites, as it can provide rapid identification of aminals that will probably have benefits from therapy. Obtained results of quantitative coprological diagnostic tests and results for anemia assessment are criteria that provide differential diagnosis between healthy and resilient animals and easier diseases diagnostic. Since resilient animals play important role in pasture contamination, the significance of their detection in understandable.

Thelazia callipaeda, originally known as an Oriental eyeworm, is a small nematode parasitizing the conjunctival sacs of domestic and wild animals and humans. Previous studies conducted in Serbia have reported the eyeworm infections in dogs, cats, and foxes, as well as in a human patient. As the data regarding thelaziosis from wildlife is still scarce, the aim of this study was to investigate the presence of T. callipaeda in gray wolf (Canis lupus) from Serbia. All collected nematodes were morphologically identified as T. callipaeda males (n = 64) or females (n = 225). Molecular characterization, conducted by PCR amplification followed by sequence analysis of partial cytochrome c oxidase subunit 1 gene (cox1), revealed only haplotype 1 of T. callipaeda. The overall prevalence of thelaziosis was 38.1% (8/21). In all positive animals, both eyes were affected, with a total parasitic load ranging from four to 132 worms per animal. Our results indicate the important epidemiological role of wolves as wildlife reservoirs of T. callipaeda, expanding geographic range of infection, as well as intra- and interspecies contact rates, although the role of other wild carnivore species (i.e., foxes and jackals) should be investigated in future studies.

This report represents the prevalence of endoparasitic infections of birds in zoological gardens in Serbia. Coprological examination conducted during 2015 and 2016, involved samples from 318 individual birds belonging to 112 species, in three zoo gardens (Belgrade, Palic and Bor). Individual faeces samples were analysed by the flotation method in salt solution. The presence of endoparasites was established in 51.96% (Belgrade), 46.16% (Palic) and 16.66% (Bor) of the tested birds. We diagnosed coccidian oocysts, eggs of Capillaria spp, Heterakis spp, Trichostrongylus spp, Ascaridia spp, Syngamus trachea and eggs of yet unidentified trematode. There were no detection of parasites from the class of cestodes. Prevalence of infection with coccidia, depending on location, was in the ranges between 8.33% and 32.84%, capillariosis 2.77% and 29.41%, heterakiosis 2.56% and 12.74%, trichostrongylidosis 5.55% and 25.64%, askaridiosis 10.25% and 10.78%, syngamosis 2.94% and trematodes of unknown etiology 2.45%. Coinfections with two or three parasitic species were detected in Belgrade and Palic Zoo Gardens, while quadruple infection was established in birds of Belgrade Zoo Garden. Obtained results can be used in establishing programs for health protection of endangered bird species.

U ovom radu ispitivana je genetička varijabilnost medonosne pčele Apis mellifera i pčelinjeg krpelja Varroa destructor, uz praćenje korelacije u distribuciji haplotipova domaćina i parazita. Za identifikaciju haplotipova pčela korišćeni su uzorci iz 48 pčelinjih društava sa osam lokaliteta na teritoriji Srbije, dok je za tipizaciju varoa i praćenje korelacije haplotipova domaćina i parazita analizirano 245 uzoraka varoe iz 29 košnica. Sekvencioniranjem tRNAleu-cox2 regiona definisano je šest haplotipova A. mellifera, pri čemu je haplotip C2aa opisan po prvi put. Zastupljenost identifikovanih haplotipova pčela razlikovala se u ukupnom uzorku, pri čemu je najzastupljeniji bio haplotip C2d, dok su najmanje prisutni bili haplotipovi C2aa i C2i. Distribucija haplotipova A. mellifera razlikovala se u zavisnosti od lokaliteta, pri čemu je na tri posmatrana lokaliteta utvrđen po jedan haplotip, na jednom lokalitetu dva i na četiri lokaliteta utvrđeno je prisustvo tri haplotipa pčela. Rezultati sekvencioniranja delova mitohondrijalnih gena V. destructor otkrili su prisustvo tačkastih mutacija na pozicijama 1932 i 10133 mtDNK unutar cox1 i cytb sekvenci. Na osnovu tih polimorfnih mesta, definisana su dva nova haplotipa varoe koji su nazvani Srbija 1 (S1) i Pešter 1 (P1). Obe otkrivene mutacije po svojoj prirodi spadaju u „tihe‟ mutacije koje ne dovode do promene proteinskog lanca. Na hromatogramima pojedinih uzoraka otkriveni su dupli pikovi na haplotip definišućim mestima unutar cox1 i cytb, čime je po prvi put utvrđena nukleotidna heteroplazmija kod vrste V. destructor. Nakon utvrđivanja polimorfnih mesta metodom sekvencioniranja, za identifikaciju haplotipova varoe i otkrivanje jedinki sa heteroplazmijom korišćene su ARMS i RFLP metode. Analizom cox1 sekvence ARMS metodom utvrđeno je 48,2% jedinki K haplotipa, 31,8% jedinki S1 haplotipa i 20,0% jedinki sa heteroplazmijom..., In this study, genetic variability of honey bee Apis mellifera and honey bee mite Varroa destructor populations was analyzed, monitoring haplotype-based distribution pattern between the host and the parasite. For the identification of honey bee haplotypes, samples from 48 honey bee colonies were used, while 245 Varroa mite samples originating from 29 colonies were analyzed for haplotype identification and monitoring correlation between A. mellifera and V. destructor haplotypes. Sequencing of tRNAleu-cox2 region revealed six A. mellifera haplotypes, with C2aa haplotype observed for the first time. Different frequency of identifed bee haplotypes was detected in total sample, with C2d haplotype being the most frequent, and C2aa and C2i haplotypes with the lowest frequency. Distribution of A. mellifera haplotypes was found to be dependent on the locality, with three sites represented by only one haplotype, one locality represented by two haplotypes and four localities represented by three honey bee haplotypes. Sequencing results of V. destructor mitochondrial fragments revealed the point mutations at position 1932 and 10133 mtDNA within cox1 and cytb sequences, respectively. Based on these polymorphic sites, two novel Varroa haplotypes were defined and named as Serbia 1 (S1) and Peshter 1 (P1). Both detected mutations were the silent ones and did not affect the protein sequence. At the sequence chromatograms in some of the analyzed samples, double peaks were detected at haplotype-defining sites within cox1 and cytb, observing the phenomenon of nucleotide heteroplasmy for the first time in V. destructor. After the polymorphic sites had been observed using the sequencing method, ARMS and RFLP assays were performed for identification of Varroa haplotypes and detection of heteroplasmic individuals. Results of the cox1 sequence analysis showed that 48,2% of individuals belonged to the K haplotype, 31,8% belonged to the S1 haplotype and 20,0% Varroa samples was found to be he

Bolesti pasa koje se prenose vektorima predstavljaju oboljenja virusne, bakterijske ili parazitske etiologije, koja se najčešće prenose hematofagnim artropodama, komarcima i krpeljima. Cilj ovog rada bio je da se ispita prisustvo infekcija čiji se uzročnici prenose komarcima (Dirofilaria immitis) i krpeljima (Borrelia burgdorferi, Anaplasma phagocytophilum/A. platys, Ehrlichia canis/E. ewingii) kod nevlasničkih pasa na teritoriji Beograda. U periodu od septembra do decembra 2015. godine sakupljeno je 86 uzoraka krvi od klinički zdravih nevlasničkih pasa iz različitih beogradskih opština. Svi uzorci testirani su modinikovanim Knott-ovim testom i komercijalnim SNAP® 4Dx Plus® Test-om. Infekcija izazvana barem jednim od ispitivanih uzročnika ustanovljena je kod 27,91% pasa. Najzastupljeniji uzročnik bila je D. immitis, koja je dijagnostikovana kod 17,44% životinja. Na prisustvo antitela protiv Anaplasma spp. pozitivno je bilo 9,30% pasa, dok je 2,33% jedinki bilo seropozitivno na B. burgdorferi, odnosno na Ehrlichia spp. Koinfekcije sa dva patogena zabeležene su kod 3,49% pasa. Rezultati ovog istraživanja pokazuju da su nevlasnički psi na teritoriji grada Beograda izloženi delovanju različitih vrsta zoonoznih uzročnika koji se prenose komarcima i krpeljima, zbog čega treba preduzeti odgovarajuće mere za suzbijanje ovih artropoda., Vector-borne diseases are caused by viruses, bacteria or parasites transmitted by hematophagous arthropods, mainly mosquitoes and ticks. The aim of this work was to study the presence of pathogens vectored by mosquitoes (DiroGilaria immitis) and ticks (Borrelia burgdorferi, Anaplasma phagocytophilum/A. platys, Ehrlichia canis/E. ewingii) in stray dogs from Belgrade. From September to December 2015, 86 blood samples were collected from apparently healthy stray dogs in different Belgrade municipalities. All samples were analysed using modiNied Knott’s test and commercial SNAP® 4Dx Plus®Test. Infection with at least one pathogen was detected in 27,91% of investigated dogs. The most prevalent agent was D. immitis that was diagnosed in 17,44% of animals. Antibodies against Anaplasma spp. were found in 9,30% of dogs and 2,33% individuals were seropositive to B. burgdorferi, as well as to Ehrlichia spp. Coinfections with two pathogens were recorded in 3,49% of dogs. Results of this research show that stray dogs in Belgrade are exposed to different zoonotic pathogens transmitted by mosquitoes and ticks, indicating the need for appropriate control against these arthropod species.

Babesiosis of domestic animals is a vector transmissible and clinically significant disease, caused by protozoa of genus Babesia and Theileria. Possible causative agents for this disease in dogs in Europe are: Babesia canis, B. gibsoni, B. vogeli and B. microti-like. Diagnostics of babesiosis of dogs was for a long time based on the visual inspection of stained blood smear under a microscope, while today there have been increasingly used molecular methods of detection in precise, species diagnostics. The objective of this work was molecular detection of the cause of babesiosis of dogs in the ticks sampled from asymptomatic dogs in the region of some Belgrade municipalities, all for better understanding of epizootiological situation. From three sites in Belgrade, there were collected 49 ticks, sampled from the dogs with no symptoms. There was carried out the determination of the ticks, and after that, DNA was isolated for molecular examination. First, theret was performed Polymerase Chain Reaction (PCR), for determining the species of the genus Babesia, and after that there was also carried out the determining of polymorphism in the length of restriction fragments (RFLP) for the purpose of the causative agent species determination. Out of the total number of the examined ticks, 18,34% were positive on Babesia spp. By RFLP method, in two cases (4,08%) B. Gibsoni was identified, while in 7 cases (14,92%) there were no restriction sites for the used enzymes, what suggests that most likely it was B. canis. The ticks positive on the cause of babesiosis were: Dermacentor reticulatus (4 cases), Rhipicephalus sanguineus (4 cases) i Ixodes ricinus (1 case). This work confirms the presence of Babesia spp. in the ticks sampled from asmptomatic dogs on the territory of Belgrade as well as the significance of PCR-RFLP method in diagnostics and identification of the causative agent of babesiosis in dogs. For the first time in Serbia, there was determined the presence of B. gibsoni, Babezioza domaćih životinja predstavlja vektorski prenosivo i klinički veoma značajno oboljenje, uzrokovano protozoama rodova Babesia i Theileria. Uzročnici koji mogu da izazovu oboljenje kod pasa u Evropi su: Babesia canis, B. gibsoni, B. vogeli i B. microti-like. Dijagnostika babezioze pasa se dugo vremena bazirala na vizuelizaciji uzročnika pregledom obojenog krvnog razmaza pod mikroskopom, dok se danas sve više koriste molekularne metode detekcije u preciznoj, specijskoj dijagnostici. Cilj ovog rada je bila molekularna detekcija različitih uzročnika babezioze pasa u krpeljima uzorkovanim sa asimptomatskih pasa na prostoru određenih beogradskih opština, radi boljeg razumevanja epizootiološke situacije. Sa tri lokacije u Beogradu je prikupljeno 49 krpelja, uzorkovanih sa pasa bez simptoma bolesti. Izvršena je determinacja krpelja, a nakon toga je izolovana DNK za molekularna ispitivanja. Prvo je urađena reakcija lančane polimerizacije (PCR) za utvrđivanje vrsta iz roda Babesia, a nakon toga i određivanje polimorfizma u dužini restrikcionih fragmenata (RFLP) u cilju specijske identifikacije uzročnika. Od ukupnog broja ispitanih krpelja 18,34% je bilo pozitivno na Babesia spp. RFLP metodom je u dva slučaja (4,08%) identifikovana B. gibsoni, dok u 7 slučajeva (14,92%) nije bilo restrkicionih mesta za korišćene enzime, što ukazuje da se najverovatnije radilo o B. canis. Pozitivni krpelji na uzročnike babezioze su bili Dermacentor reticulatus (4 slučaja), Rhipicephalus sanguineus (4 slučaja) i Ixodes ricinus (1 slučaj). Ovaj rad potvrđuje prisustvo Babesia spp. u krpeljima uzorkovanim sa asimptomatskih pasa na teritoriji grada Beograda, kao i značaj PCR-RFLP metode u dijagnostici i identifikaciji uzročnika babezioze pasa. Prvi put u Srbiji je utvrđeno prisustvo B. gibsoni kod krpelja (vrsta Rhipicephalus sanguineus).

In this study, genetic variability of honey bee Apis mellifera and honey bee mite Varroa destructor populations was analyzed, monitoring haplotype-based distribution pattern between the host and the parasite. For the identification of honey bee haplotypes, samples from 48 honey bee colonies were used, while 245 Varroa mite samples originating from 29 colonies were analyzed for haplotype identification and monitoring correlation between A. mellifera and V. destructor haplotypes. Sequencing of tRNAleu-cox2 region revealed six A. mellifera haplotypes, with C2aa haplotype observed for the first time. Different frequency of identifed bee haplotypes was detected in total sample, with C2d haplotype being the most frequent, and C2aa and C2i haplotypes with the lowest frequency. Distribution of A. mellifera haplotypes was found to be dependent on the locality, with three sites represented by only one haplotype, one locality represented by two haplotypes and four localities represented by three honey bee haplotypes. Sequencing results of V. destructor mitochondrial fragments revealed the point mutations at position 1932 and 10133 mtDNA within cox1 and cytb sequences, respectively. Based on these polymorphic sites, two novel Varroa haplotypes were defined and named as Serbia 1 (S1) and Peshter 1 (P1). Both detected mutations were the silent ones and did not affect the protein sequence. At the sequence chromatograms in some of the analyzed samples, double peaks were detected at haplotype-defining sites within cox1 and cytb, observing the phenomenon of nucleotide heteroplasmy for the first time in V. destructor. After the polymorphic sites had been observed using the sequencing method, ARMS and RFLP assays were performed for identification of Varroa haplotypes and detection of heteroplasmic individuals. Results of the cox1 sequence analysis showed that 48,2% of individuals belonged to the K haplotype, 31,8% belonged to the S1 haplotype and 20,0% Varroa samples was found to be he, U ovom radu ispitivana je genetička varijabilnost medonosne pčele Apis mellifera i pčelinjeg krpelja Varroa destructor, uz praćenje korelacije u distribuciji haplotipova domaćina i parazita. Za identifikaciju haplotipova pčela korišćeni su uzorci iz 48 pčelinjih društava sa osam lokaliteta na teritoriji Srbije, dok je za tipizaciju varoa i praćenje korelacije haplotipova domaćina i parazita analizirano 245 uzoraka varoe iz 29 košnica. Sekvencioniranjem tRNAleu-cox2 regiona definisano je šest haplotipova A. mellifera, pri čemu je haplotip C2aa opisan po prvi put. Zastupljenost identifikovanih haplotipova pčela razlikovala se u ukupnom uzorku, pri čemu je najzastupljeniji bio haplotip C2d, dok su najmanje prisutni bili haplotipovi C2aa i C2i. Distribucija haplotipova A. mellifera razlikovala se u zavisnosti od lokaliteta, pri čemu je na tri posmatrana lokaliteta utvrđen po jedan haplotip, na jednom lokalitetu dva i na četiri lokaliteta utvrđeno je prisustvo tri haplotipa pčela. Rezultati sekvencioniranja delova mitohondrijalnih gena V. destructor otkrili su prisustvo tačkastih mutacija na pozicijama 1932 i 10133 mtDNK unutar cox1 i cytb sekvenci. Na osnovu tih polimorfnih mesta, definisana su dva nova haplotipa varoe koji su nazvani Srbija 1 (S1) i Pešter 1 (P1). Obe otkrivene mutacije po svojoj prirodi spadaju u „tihe‟ mutacije koje ne dovode do promene proteinskog lanca. Na hromatogramima pojedinih uzoraka otkriveni su dupli pikovi na haplotip definišućim mestima unutar cox1 i cytb, čime je po prvi put utvrđena nukleotidna heteroplazmija kod vrste V. destructor. Nakon utvrđivanja polimorfnih mesta metodom sekvencioniranja, za identifikaciju haplotipova varoe i otkrivanje jedinki sa heteroplazmijom korišćene su ARMS i RFLP metode. Analizom cox1 sekvence ARMS metodom utvrđeno je 48,2% jedinki K haplotipa, 31,8% jedinki S1 haplotipa i 20,0% jedinki sa heteroplazmijom...

Blood smears stained with Diff Quick are the initial tool for cytological diagnosis of Haemogregarina spp. However, the development of sensitive and specific molecular methods enabled the detection and identification of parasites in the sample and to clarify the evolutionary relationships of adeleorinid parasites within the Apicomplexa. The current study was attempted in order to perform cytological investigation and molecular identification of the hemoparasites in thirty European pond turtles (Emys orbicularis) from the quarantine section at Belgrade Zoo, which have been found in poor health condition with massive skin hemorrhages, based on intraerythrocytic parasitic forms on hematological smears and 18S rDNA sequence, respectively. Different life cycle stages of the Haemogregarina sp. were noticed within the erythrocytes in the peripheral blood. Biochemical analysis indicated lower values of AST and iron in most of the infected turtles while hematological analysis showed a changed hematocrit value, a decrease in the number of red blood cells and low hemoglobin levels. Amplifications of the 18S rDNA sequence of Haemogregarina were detected in 30/30 (100%) turtles with clinical symptoms. The identity of PCR products was confirmed by direct DNA sequencing. Future research concerning H. stepanowi in Serbia should be applied to its definitive host-the leech.

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 findings 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 confirmed 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 confirmation 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)., Pčelinji krpelj Varroa destructor i mikrosporidija Nosema ceranae su najznačajniji faktori rizika po zdravlje pčela i pčelarstvo. Dugo se smatralo da N. apis i N. ceranae inficiraju isključivo ćelije crevnog epitela pčela, međutim neki nalazi ukazuju na prisustvo ovih mikrosporidija i u drugim tkivima. Postoje indikacije da se ovi patogeni mogu naći i u hemolimfi pčela, kao medijumu za njihovu distribuciju do anatomski udaljenih tkiva. S obzirom na to da se V. destructor kao ektoparazit hrani hemolimfom pčela, cilj ovog rada bio je ispitivanje prisustva DNK mikrosporidija roda Nosema u hemolimfi pčela i pčelinjem krpelju V. destructor. Istraživanje je obavljeno na uzorcima pčelinje hemolimfe i pčelinjim krpeljima V. destructor iz 44 društva Apis mellifera. Iz svake košnice je uzorkovano pet krpelja i 10 μL hemolimfe (sakupljene iz 4-5 pčela) koji su korišćeni kao uzorci za izolaciju DNK i ispitivanje prisustva Nosema spp. putem PCR metode. Prisustvo DNK N. ceranae je potvrđeno u 61,36% analiziranih krpelja V. destructor i 68,18% uzoraka hemolimfe pčela, što predstavlja prvu detekciju prisustva DNK N. ceranae u hemolimfi pčela i krpeljima V. destructor. Nalaz DNK N. ceranae u V. destructor može se tumačiti kao rezultat ishrane V. destructor hemolimfom pčela koja je inficirana sa N. ceranae. Međutim, da bi se potvrdila vektorska uloga V. destructor u širenju nozemoze, potrebna su dalja mikroskopska istraživanja radi detekcije spora u oba ispitivana medijuma (hemolimfi i unutrašnjim tkivima V. destructor krpelja).

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., Thelazia callipaeda je parazitska nematoda koja izaziva infekciju oka različitih vrsta sisara i ljudi, a koja se klinički manifestuje pojačanim suzenjem, pojavom konjuktivitisa, keratitisa, zamućenja rožnjače ili pojave ulcera na rožnjači. U ovom radu opisani su nalazi autohtone telazioze pasa i mačaka na šest različitih lokaliteta u Srbiji. Iz očiju inficiranih životinja (n=6) sa simptomima uni- ili bilateralnog konjuktivitisa sakupljeno je 285 parazita (85 mužjaka i 200 ženki), pri čemu se broj parazita po životinji kretao od 7 do 150. Svi paraziti morfološki su identifikovani kao T. callipaeda, dok su molekularne analize citohrom oksidaza 1 (cox1) gena utvrdile postojanje h1 haplotipa, koji je do sada i jedini utvrđeni haplotip ovog parazita u Evropi. S obzirom da se radi o novootkrivenom parazitu na teritoriji Srbije koji može inficirati i ljude, neophodno je uložiti napore u edukaciju vlasnika životinja i saradnju stručnih službi u cilju kontrole ove zoonoze.

A coprological examination of 680 grazing sheep was performed in Eastern Serbia from March 2011 to November 2012 in order to determine the presence of gastrointestinal (GI) nematode parasites. Fecal samples were randomly collected and examined by using qualitative and quantitative coprological techniques. It was found that 74.56% sheep were infected. Samples that contained nematode eggs were processed for larval development and eleven nematode genera were identified: Haemonchus (46.91%), Ostertagia (25.88%), Marshallagia (21.91%), Cooperia (14.12%), Trichostrongylus (39.85%), Nematodirus (35.88%), Bunostomum (23.97%), Strongyloides (17.06%) Oesophagostomum (40.73%), Chabertia (32.79%) and Trichuris (10.88%). Higher prevalence of infection was observed in females (p lt 0.01), as well as in adults (p lt 0.001). Regarding the intensity of infection, in 40.63% sheep it was low, in 51.87% moderate and in 7.50% high. There was no difference in intensity of infection considering sex and age of animals. Moreover, simultaneous infection with different number of nematode genera was dependent on sheep's age (p lt 0.001). These results suggest that GI nematodes are a conspicuous problem of grazing sheep in the study area., U periodu od marta 2011. do novembra 2012. godine, na teritoriji Istočne Srbije sprovedeno je koprološko ispitivanje 680 ovaca držanih u pašnom načinu gajenja na prisustvo gastrointestinalnih (GI) nematoda. Uzorci fecesa uzimani su metodom slučajnog uzorka i ispitivani korišćenjem kvalitativnih i kvantitativnih koproloških tehnika. Ispitivanjem je utvrđeno 74,56% inficiranih ovaca. Uzorci u kojima su pronađena jaja nematoda podvrgnuti su determinaciji larvi, pri čemu je identifikovano jedanaest rodova nematoda: Haemonchus (46,91%), Ostertagia (25,88%), Marshallagia (21,91%), Cooperia (14,12%), Trichostrongylus (39,85%), Nematodirus (35,88%), Bunostomum (23,97%), Strongyloides (17,06%) Oesophagostomum (40,73%), Chabertia (32,79%) i Trichuris (10,88%). Veća prevalencija infekcije uočena je kod ženki (p lt 0,01), kao i kod adultnih životinja (p lt 0,001). Kod 40,63% ovaca, infekcija je bila niskog, kod 51,87% umerenog, a kod 7,50% visokog intenziteta. Nisu dokazane razlike u intenzitetu infekcije u zavisnosti od pola i starosti životinja. Istovremena infekcija sa više rodova nematoda zavisila je od starosti ovaca (p lt 0,001). Dobijeni rezultati upućuju na zaključak da su GI nematode značajan problem ovaca držanih na pašnjaku u ispitivanom području.

The incidence, symptoms and consequences of Nosema ceranae infection were monitored in 200 honey bee colonies in Serbia over 5 years (2008-2012) to reveal if they display characteristics described for nosemosis type C. Continual high frequency of N. ceranae-positive colonies was recorded within each study year, ranging from 73 to 98 %. A seasonal pattern in N. ceranae incidence was observed over 4 years (2008-2011). Symptoms traditionally attributed to Nosema apis infection were observed in the majority of N. ceranae-infected colonies, both among surviving and those that died. These symptoms could not serve as predictive markers for colony losses, as they were significantly more frequently observed among surviving colonies in 2010, 2011 and 2012. No clear association between N. ceranae infection and colony losses was affirmed neither during the winter nor during the summer season. Thus, N. ceranae infection in Serbian bees do not fit the characteristics for nosemosis type C.

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. Varroa destructor (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.

This study was undertaken to estimate the prevalence of gastrointestinal helminths in calves in western Serbia. Throughout 2011 faecal samples were collected from 600 calves aged up to 180 days, samples were examined with the flotation method and a modified McMaster technique. The parasitizing helminth species were identified and the level of infection compared between different age groups. As many as 64.17% animals were found to be infected. The following parasite species were diagnosed: Moniezia spp. (3.17%), Toxocara vitulorum (35.00%), Strongyloides papillosus (34.50%), gastrointestinal strongyles (4.50%) and Trichuris discolor (2.17%). The majority of calves were infected with two, fewer with three or one helminth species, and the smollest number of calves harboured four parasite species. The prevalence of established helminth infections varied depending on the calves' age., Studija je sprovedena sa ciljem da se utvrdi prevalencija gastrointestinalnih helminata kod teladi u zapadnom delu Srbije. Tokom 2011. godine prikupljeni su uzorci fecesa ukupno 600 teladi starosti do 180 dana. Uzorci su pregledani metodom flotacije i modifikovanom metodom po McMasteru. Determinisane su vrste helminata i određena je prevalencija infekcije kod teladi različite starosti. Rezultati ispitivanja pokazali su da je infekcija helmintima bila prisutna kod 64,17% pregledanih životinja. Ustanovljene su sledeće vrste helminata: Moniezia spp. (3,17%), Toxocara vitulorum (35,00%), Strongyloides papillosus (34,50%), želudačnocrevne strongilide (4,50%) i Trichuris discolor (2,17%). Većina teladi istovremeno je bila inficirana dvema vrstama, zatim sa tri i jednom vrstom helminta, dok je kod samo nekoliko životinja bilo ustanovljeno prisustvo četiri vrste ili grupe helminata. Prevalencija infekcija helmintima razlikovala se kod teladi različite starosti.