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Blastocystis is a common protist colonizing the gastrointestinal tract of humans and various animals. The first subtyping of Blastocystis isolates in pigs and humans in Serbia revealed unusual avian-specific subtype ST6 in humans. In total, 48 pig faecal specimens collected on seven pig farms and 50 human faecal specimens positive to Blastocystis by microscopic examination were selected for the study. Eleven randomly selected PCR-positive pig samples and 10 samples from human patients (with gastrointestinal complaints) were subjected to SSU rDNA sequencing. Three subtypes were identified (ST3, ST5 and ST6) by phylogenetic analysis. ST5 was found in all pig samples; while in human samples, we detected ST3 and ST6. The latter subtype is relatively uncommon in Europe and highly adapted to avian hosts; therefore, the possibility of sporadic zoonotic transmission to human patients should not be ignored.

The objective of this paper is to describe two cases of heartworm disease in dogs presented with ascites and dyspnea. A diagnosis of mixed infection with both Dirofilaria immitis (D. immitis) and Dirofilaria repens (D. repens) was established in the first case, while D. immitis was detected in the other dog. Based on clinical, parasitological, laboratory and radiographic examinations, the diagnosis of severe stage heartworm disease (HWD) was established because both dogs were in heart failure due to HWD. An alternative therapeutic protocol with combined doxycycline and ivermectin was used as an antiparasitic treatment in both cases. No microfilariae were detected after 30 and 60 days, and no antigen to D. immitis was detected after 8 and 12 months in the first and second case, respectively. Based on the clinical examinations and parasitological testing performed, the two dogs were considered cured of HWD by this alternative therapy.

Lyme borreliosis is the most common tick-borne infectious disease in Eurasia. Borrelia miyamotoi is the only known relapsing fever Borrelia group spirochete transmitted by Ixodes species. The aim of this study was to investigate the presence of Lyme Borrelia spp. and relapsing fever Borrelia spp. in Ixodes ricinus ticks collected from dogs and the vegetation from different parts of Vojvodina, Serbia. A total of 71 Ixodes ricinus ticks were collected and screened for the presence of Lyme Borrelia spp. group and relapsing fever Borrelia spp. by real-time PCR for the Borrelia flagellin B (flaB) gene followed by DNA sequencing of PCR products. Species identification was verified by PCR of the outer surface protein A (ospA) gene for Lyme Disease Borrelia spp. and by PCR of the glycerophosphodiester phosphodiesterase (glpQ) gene for relapsing fever Borrelia spp. Lyme Borrelia spp. were found in 15/71 (21.13%) of the ticks evaluated and included B. luisitaniae (11.3%), B. afzelii (7%), B. valaisiana (1.4%), and B. garinii (1.4%). Borrelia miyamotoi, from the relapsing fever Borrelia complex, was found, for the first time in Serbia, in one (1.4%) nymph collected from the environment. Co-infections between Borrelia species in ticks were not detected. These results suggest that the dominance of species within B. burgdorferi s.l. complex in I. ricinus ticks may vary over time and in different geographic regions. Further systematic studies of Borrelia species in vectors and reservoir hosts are needed to understand eco-epidemiology of these zoonotic infections and how to prevent human infection in the best way.

Ticks play an important role in disease transmission globally due to their capability to serve as vectors for human and animal pathogens. The Republic of Serbia is an endemic area for a large number of tick-borne diseases. However, current knowledge on these diseases in Serbia is limited. The aim of this study was to investigate the presence of new emerging tick-borne pathogens in ticks collected from dogs and the vegetation from different parts of Vojvodina, Serbia. A total of 187 ticks, including 124 Rhipicephalus sanguineus, 45 Ixodes ricinus and 18 Dermacentor reticulatus were collected from dogs. In addition, 26 questing I. ricinus ticks were collected from the vegetation, using the flagging method, from 4 different localities in Vojvodina, Serbia. DNA was extracted from each tick individually and samples were tested by either conventional or real-time PCR assays for the presence of Rickettsia spp.-DNA (gltA and ompA gene fragments), Ehrlichia/Anaplasma spp.-DNA (16S rRNA gene fragment) and Hepatozoon spp./Babesia spp.-DNA (18S rRNA gene fragment). In addition, all I. ricinus DNA samples were tested for Bartonella spp.-DNA (ITS locus) by real-time PCR. In this study, the presence of novel emerging tick-borne pathogens including Rickettsia raoultii, Rickettsia massiliae, Babesia venatorum, Babesia microti, Hepatozoon canis and Candidatus Neoehrlichia mikurensis was identified for the first time in Serbia. Our findings also confirmed the presence of Rickettsia monacensis, Babesia canis and Anaplasma phagocytophilum in ticks from Serbia. The findings of the current study highlight the great diversity of tick-borne pathogens of human and animal importance in Serbia. Physicians, public health workers and veterinarians should increase alertness to the presence of these tick-borne pathogens in this country.

Seroprevalence of tick-borne infections in endemic areas could be high. In this study, we investigated the seroprevalence of tick-borne pathogens (Anaplasma phagocytophilum, Borrelia burgdorferi, Babesia canis, Ehrlichia canis, Rickettsia conorii) in hunting dogs, naturally infected with one or more pathogens. Serological test results of the investigated animals were compared to those from clinical examination, as well as from haematological and biochemical analyses. A total of 74.14% dogs were seropositive (R.conorii 44.83%, B. canis 32.76%, B. burgdorferi 25.86%, E. canis 13.79%, A. phagocytophilum 8.47%), with 25.86% of dogs seropositive to two pathogens, 15.52% seropositive to three pathogens, and 1.72% of dogs seropositive to four pathogens. Among all registered clinical signs, only pyrexia (p

Tick-borne encephalitis (TBE) is a zoonotic flaviviral infection that is a growing public health concern in European countries. The aims of this research were to detect and characterize tick-borne encephalitis virus (TBEV) in Ixodes ricinus ticks at presumed natural foci in Serbia, and to determine seroprevalence of TBEV IgG antibodies in humans and animals. A total of 500 I. ricinus ticks were examined for the presence of TBEV by real-time RT-PCR, and conventional nested PCR and sequencing. To determine TBEV seroprevalence, 267 human sera samples were collected, as were 200 sera samples from different animal species. All sera samples were examined by ELISA for the presence of anti-TBEV antibodies. To exclude cross-reactivity, all sera samples were tested for anti-West Nile virus (WNV) antibodies and all human sera samples were also tested for anti-Usutu virus antibodies by ELISA. Results of this preliminary study indicated TBEV activity in Serbia at two microfoci. Several decades after the previous documentation of TBEV in Serbia, we have demonstrated the presence of TBEV in I. ricinus questing nymphs (prevalence 2% and 6.6% at the two different localities) and anti-TBEV antibodies in humans (seroprevalence 0.37%). Moreover, we show for the first time TBEV seroprevalence in several animal species in Serbia, including dogs (seroprevalence 17.5%), horses (5%), wild boars (12.5%), cattle (2.5%), and roe deer (2.5%). None of the goats tested was positive for anti-TBEV IgG antibodies. TBEV isolate from I. ricinus tick in this study belonged to the Western European subtype. To understand the true public health concern in Serbia, detailed epidemiological, clinical, virological, and acarological research are required. This is important for implementation of effective control measures to reduce the incidence of TBE in Serbia.

The object of this study was to determine the effect of the area available per piglet, during the nurturing phase, on production performance and presence of Salmonella spp. Testing was performed on two farrow-to-finish farms over a period of two years. The study was carried out in three variants: 0.16 m 2 , 0.25 m 2 and 0.32 m 2 of floor space per piglet. Pooled faeces samples, which were tested for the presence of Salmonella spp., were collected three times during the nurturing stage: 7, 21 and 35 days after weaning. The piglets which had the lowest spatial area available (0.16 m 2 ) had average daily gain (ADG) significantly lower (Po0.05), and feed conversion ratio (FCR) higher than piglets with 0.25 m 2 and 0.32 m 2 of floor space available. Morbidity and mortality were significantly higher in piglets with 0.16 m 2 of floor space available, compared to piglets housed on larger floor spaces, for both farms examined. The overall presence of Salmonella spp. was 6.2% (19/ 306). Out of 19 Salmonella-positive faecal samples, 15 were from piglets with 0.16 m 2 of floor space available (78.9%). The results showed that holding piglets in an area smaller than recommended has no economic justification and may imposes a higher risks of infection of piglets.

The aim of this research was to determine the influence of immunostimulants to the height of antibody titers specific to PCV2 after vaccination of piglets, based on monitoring the class G antibody titers in piglets and fattening pigs. The experiment was conducted on 60 piglets, divided into two groups of 30 ones each. Both groups were vaccinated on the 21st day of age, and to the second group (group B) immunostimulant was added in food. Three weeks after the vaccination had been performed in the experimental groups, average titer in the population of piglets in group B was 10% higher compared to the average titer in group A, and 7 weeks after the vaccination, average titer values were also higher in group B (EU 1589) compared to group A (1440 EU). Homogeneous immune response and higher average titer values of antibodies against PCV2 after vaccination in group B were probably the result of the use of immuniostimulant containing RNA, nucleotides and nucleotide precursors that are necessary for the synthesis of antibodies.

Ticks are vectors for many infectious diseases and represent a constant threat to human population and other animals, especially with respect to zoonoses. Th e cause of granulocyte anaplasmosis Anaplasma phagocytophilum is a gram-negative, obligate intracellular bacterium that infects people as well as various domestic and wild animals. Th e agent is spread worldwide, persisting in the natural environment through an enzootic circle between ticks and their hosting vertebrae. Th e aim of this paper is to demonstrate the distribution of ticks’ infection with Anaplasma phagocytophilum in the territory of the Autonomous Province of Vojvodina. Ticks were collected at ten locations in the Autonomous Province of Vojvodina, Serbia, which are a rural habitat for ticks. By applying the nested PCR method, the ticks were examined for the presence of specifi c DNA p44/msp2 Anaplasma phagocytphilum. Of the ten examined pools of ticks collected in the area of the Autonomous Province of Vojvodina, the presence of genome of the agent Anaplasma phagocytophilum was confi rmed in six pools applying the PCR method. In five pools of ticks of the species Ixodes ricinus, presence of agent Anaplasma phagocytophilum was confi rmed. Th ese ticks were found at the following locations: Poplar Research Institute (2 locations), Fruška gora (2 locations) Poloj-forest Bačka Palanka (1 location). In one pool of ticks of the species Dermacentor reticulatus from the location Poloj-forest Bačka Palanka, we confi rmed the presence of Anaplasma phagocytophilum genome. This infection can be a problem for public health, so further and more comprehensive acharological and epidemiological research is necessary in the Autonomous Province of Vojvodina.

Lyme disease is a multisystemic disease, zoonotic in nature, caused by the Borrelia burgdorferi sensu lato complex. In the continent of Europe, these spirochetes are predominantly transmitted by ticks of the genus Ixodes. Small mammals and birds have particular significance as reservoirs of the cause of lyme disease. The objective of these epidemiological investigations was to determine the value of IgG seroprevalence to Borrelia burgdorferi and to secure the geographic distribution of seropositive dogs in Vojvodina. The investigations covered 135 dogs that were not vaccinated against lyme disease. The indirect ELISA test was used to determine IgG prevalence to Borrelia burgdorferi antigens. Reactive blood serums of dogs were tested again using the rapid immunochromatographic and immunoblot test. A seroprevalence of G class antibodies to antigens of lyme disease causes of 8.1% (11/135) was established in the examined dog population of Vojvodina. The biggest number of positive results was recorded for the South Backa District. The presented value for the seroprevalence of anti-Borrelia burgdorferi antibodies in the dog population indicates the exhistence of a significant risk of humans becoming infected with the cause of lyme disease in Vojvodina.

Dermatophytoses are frequent contagious fungal skin diseases that affect the skin of people and animals. Zoophile dermatophytes pose a significant problem for both human and veterinary medicine, and they are especially present among bovines. In this paper we showed a simultaneous occurrence of trichophytosis among professionally exposed people and bovines on a farm in Vojvodina, Serbia. The tested samples (skin scrapings and hair) originating from people and animals, were positive for Trichophyton verrucosum dermatophyte which was determined by applying a direct microscopic examination of the smears, as well as the isolation and identification of the agents.

Sow mortality is a great challenge in intensive pig production worldwide. The aim of this study was to forensically examine the reason of sow death for a two-year period on two farms with intensive pig keeping, based on available data. In sows with a greater number of farrowings (over VII) greater number of deaths was found. Seasonality has an impact on sow mortality, in the summer period a larger number of sow death was found. Poor sow condition that usually occurs in the second half of lactation and after weaning, are the predisposition to the factors that lead to the sow death. On both examined farms in more than 60% of dead sows Clostridium spp. and Escherichia coli were isolated. In order to reduce the mortality of sows more attention should be paid to the older sows with a larger number of farrowings, provide better conditions in summer, cooling, and pay more attention to sows during the period from farrowing to the next insemination, respectively.

Canine monocytic ehrlichiosis is a bacterial, vector-transmitted infection caused by Ehrlichia canis. Th e pathogen is mainly transmitted by ticks Rhipicephalus sanguineus. The disease highest distributionis most widely distributed in tropical and subtropical countries but it is also reported in Mediterranean countries of Europe (Spain, France, Italy, Turkey). Temperate continental climate and presence of these tick species are responsible for the maintenance and spreading of canine monocytic ehrlichiosis within the dog population in our region as well. Since hunting dogs are more exposed to tick bites then pet dogs, ed thisour study was conducted with the aims of determining the seroprevalence and basic epidemiological characteristics of monocytic ehrlichiosis infection in the population of hunting dogs, and comparing the obtained results with the results of other authors. This research involved 58 hunting dogs from the region of Autonomous Province of Vojvodina. All dogs were clinically examined and their basic epidemiological characteristics were recorded. Then, blood samples were collected in order to determine the presence of specifi c G class antibodies against Ehrlichia canis antigens. An indirect immunofl uorescence test manufactured by VMRD, U.S.A., was used. In this study, the seroprevalence of monocytic ehrlichiosis in a population of examined hunting dogs from the region of Vojvodina was 13.79%. Th is rate is similar to the seroprevalence of monocytic ehrlichiosis in the general population of dogs in Vojvodina.

The aim of this research was to determine the effect of vaccination on the amount of antibody titers specific for PCV2, and to determine the effect of vaccination on characteristics of pig production. The first group (A) was vaccinated at 15 days old, the second (B) at 21 days old while the third (C) was the control group. Group B piglets attained the best results, so the vaccination of piglets at 21 days old would have an advantage compared to vaccination at 15 days old, although we note that at 15 days old, there is a far greater influence of maternal antibodies on the creation and development of immune responses in the piglets after vaccination.

Borrelia burgdorferi sensu lato (s.l.) complex represents a group of different types of spirochete that are present globally, which causes Lyme borreliosis. The total number of types is still not final because new genetic and antigenic isolates are still being described. The clinical picture of Lyme borreliosis in people is polymorphic and is characterized with symptoms similar to flu syndromes together with erythema migrans, rheumatologic, cardiologic and neurological complications. Apart from people, dogs, horses, bovines and sheep can suffer from Lyme borreliosis. In Europe, the most important vector transmitting Lyme disease is the tick Ixodes ricinus. A description of different species of Borrelia in ticks has opened a completely new field of investigating the ecology of Lyme borreliosis. It is especially important to investigate a connection between different types of Borrelia, ticks as vectors and vertebrates as the reservoirs on various geographical localities. The aim of the work is to isolate Borrelia species from the collected ticks I. ricinus from the geographical territory of Vojvodina, and to do a molecular characterization of the isolated strains of Borrelia. A total of 12 tick pools of the type I. ricinus were cultivated in Barbour-Stoenner-Kelly-H medium with additional antibiotics and subcultivated into a modified Kelly-Pettenkofer/Preac-Mursic medium, until a clear culture is reached. For the molecular characterization of isolated strains of Borrelia, molecular methods MluI-LRFP and real-time PCR for hbb gene were used. In this research, out of 12 pools of ticks species I. ricinus 3 strains of Borrelia from the B. burgdorferi s.l. complex were isolated. All three isolates of Borrelia from ticks of the tick species I. ricinus from the territory of Vojvodina were identified as Borrelia afzelii by applying molecular methods (MluI-LRFP and real-time PCR for hbb gene). By applying MluI-LRFP all three isolated strains of B. afzelii were characterized as a subtype Mlal. In this research, like in the previous researches, we have not proved the presence of a pathogenic species Borrelia spielmanii in ticks of the species I. ricinus.