One health approach to study human health risks associated with Dermanyssus gallinae mites.

Despite the significant health risks associated with Dermanyssus gallinae infestations in humans, they are often overlooked. This study investigated a household case of D. gallinae infestation and explored the resulting clinical manifestations and risk of infection in family members. Microfluidic PCR was employed for high-throughput screening of pathogens in collected mites and blood samples from both chickens and family members. Morphological and molecular examinations confirmed the identity of the mites as D. gallinae sensu stricto (s.s.), with evidence indicating recent blood feeding. Results indicated that the mites exclusively harbored various pathogens, including Bartonella spp., Ehrlichia spp., Apicomplexa, and Theileria spp. Blood samples from family members and poultry tested negative for these pathogens, suggesting a potential reservoir role for D. gallinae. The study further identified haplotypes of D. gallinae, classifying them into D. gallinae s.s., cosmopolitan haplogroup A. Serological analysis revealed elevated IgE seroreactivity against mite proteins in the family member with bite lesions. Antibodies against Bartonella spp. were detected in this individual, indicating exposure to the pathogen. In summary, this study sheds light on the clinical manifestations, pathogen detection, and genetic characterization of D. gallinae infestations, underscoring the necessity of adopting comprehensive approaches to manage such infestations effectively.

Real-world evidence of rabies post-exposure prophylaxis in Serbia: Nation-wide observational study (2017-2019).

BACKGROUND: Rabies remains a deadly zoonotic disease, primarily prevalent in Eastern European countries, with a significant global burden in Asia and Africa. Post-exposure prophylaxis (PEP) is critical to prevent clinical rabies. Serbia, a country with a relatively low animal rabies incidence, has been implementing a 4-dose Essen PEP regimen for 13 years. This real-world study aimed to assess the effectiveness of the 4-dose Essen regimen, considering demographic and clinical factors, after WHO Category III exposure. METHOD: The study included 601 patients who received the 4-dose Essen PEP and 79 who received an additional 5th dose. RESULTS: Age emerged as a critical factor influencing seroconversion rates after the 4-dose regimen, with older individuals exhibiting lower RVNA titers. Logistic regression indicated a 3.18% decrease in seroconversion odds for each added year of age. The Cox proportional hazards mixed model highlighted age-related risks, with age groups 45-60 and 75-92 at the highest risk of non-seroconversion. Human Rabies Immune Globulin (HRIG) administration was associated with lower RVNA values after the 4-dose regimen, suggesting interference with vaccine immunogenicity among people who received larger doses of HRIG. CONCLUSIONS: This study provides valuable real-world evidence for rabies PEP in a non-homogeneous population with potential comorbidities. The results underscore the importance of optimizing PEP strategies, particularly in older individuals, and reconsidering HRIG dosing to improve seroconversion rates.

Evidence of tick-borne encephalitis virus neutralizing antibodies in Serbian individuals exposed to tick bites.

Introduction: Tick-borne encephalitis (TBE) is an emerging vector-borne and food-borne disease caused by the tick-borne encephalitis virus (TBEV; Orthoflavivirus encephalitidis), with a distribution spanning the Eurasian continent. Despite its significant public health impact in various European regions, TBE remains largely underdiagnosed in Serbia due to limited awareness and diagnostic challenges. In response to this, our study aimed to comprehensively assess TBEV exposure in individuals infested with ticks and to identify potential TBEV foci within Serbia. Materials and methods: From 2019 to 2021, we conducted an observational study involving 450 patients who reported tick infestations. Results: Our demographic analysis revealed a median age of 38 years, with a slight male predominance among the participants. We documented tick infestations in 38 municipalities across 14 districts of Serbia, with a notable concentration in proximity to Fruska Gora Mountain. The ticks most frequently removed were Ixodes ricinus, with nymphs and adult females being the predominant stages. On average, nymphs were removed after about 27.1 hours of feeding, while adult females remained attached for approximately 44.4 hours. Notably, we found age as a significant predictor of infestation time for both nymphs and adult females. Furthermore, we detected TBEV-neutralizing antibodies in 0.66% of the serum samples, shedding light on potential TBEV foci, particularly in Fruska Gora Mountain and other regions of Serbia. Conclusion: Our study emphasizes the urgent need for active TBE surveillance programs, especially in areas suspected of hosting TBEV foci, in order to assess the true TBE burden, identify at-risk populations, and implement effective preventive measures.

Tick-Borne Encephalitis Virus and Borrelia burgdorferi Seroprevalence in Balkan Tick-Infested Individuals: A Two-Centre Study.

Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are important tick-borne diseases in Europe. This study aimed to investigate the seroreactivity against Borrelia burgdorferi and TBE virus (TBEV) in tick-infested individuals in North Macedonia and Serbia. Serum samples were collected from tick-infested individuals and from healthy individuals in the same regions. Samples were tested for anti-Borrelia IgG reactivity and TBEV-neutralizing antibodies. Results showed higher seroreactivity against Borrelia antigens in patients and healthy donors from Novi Sad compared to those from the Skopje region. However, there was no statistically significant difference between tick-infested patients and healthy donors within each region. No TBEV-neutralizing antibodies were detected in participants from Novi Sad or in the control groups, except for one person from North Macedonia who had a moderate TBEV-neutralizing reaction. The study highlights the need for improved surveillance and diagnostic capabilities for LB and TBE in these regions. It also suggests the potential existence of TBEV foci in North Macedonia. The findings provide a complementary understanding of the LB and TBE epidemiology in the studied regions; however, further research is needed to investigate the presence and distribution of Borrelia spp. and TBEV in ticks to assess the significance of detected seroreactivity.

Emerging tick-borne spotted fever group rickettsioses in the Balkans.

The impact of tick-borne pathogens (TBPs) on human health has increased in the last decades, since the incidence of emerging and re-emerging infectious and zoonotic tick-borne diseases has increased worldwide. Tick-borne rickettsiae of the Spotted Fever group (SFGR) are considered as emerging pathogens that can infect humans and cause a variety of non-specific clinical symptoms. Here, we report nine cases of atypical tick-borne diseases (9/460; 1.95%) that occurred over a period of four months (from 15 April 2021 to 16 August 2021) in Serbia, from which five cases were classified as confirmed SFGR infection, two cases as probable SFGR infection and two cases as suspected SFGR infection. Within cases of confirmed SFGR infection, R. helvetica was detected as the causative agent in two cases. The most common clinical finding was non-expanding persistent circular redness, followed by eschar and enlargement of regional lymph nodes, and pain at lesion site. Rickettsia outer membrane protein B (ompB) and citrate synthase (gltA) gene fragments were amplified from clinical samples and ticks attached to patients and IgG reacting with Rickettsia conorii antigen were detected in sera samples of patients, which are highly suggestive of exposure to SFGR. Surveillance and monitoring of rickettsial diseases in Serbia should continue and extended to new areas due to the increasing trend of clinical infections caused by SFGR in the country.

Differential detection of tick-borne pathogens in human platelets and whole blood using microfluidic PCR.

The tick-borne pathogens (TBPs) with adhesive phenotype can use platelets for dissemination and colonization of distant tissues and organs, and it has been shown that they can be found concentrated in the platelet fraction of blood. This study shows the differential presence of TBPs in samples of human platelet fraction (n = 68), whole blood samples (n = 68) and ticks collected (n = 76) from the same individuals, using an unbiased high-throughput pathogen detection microfluidic system. The clinical symptoms were characterized in enrolled patients. In patients with suspected TBP infection, serological assays were conducted to test for the presence of antibodies against specific TBPs. Tick species infesting humans were identified as Ixodes ricinus, Dermacentor reticulatus, and Haemaphysalis punctata. Eight patients developed local skin lesions at the site of the tick bite including non-specific lesions, itching sensation at the lesion site, and eschar. Most common TBPs detected in platelet fraction were Borrelia spielmanii and Rickettsia sp., followed by Borrelia afzelii and Anaplasma phagocytophilum. Multiple infections with three TBPs were detected in platelet fraction. In whole blood, most common TBPs detected were Anaplasma spp. and A. phagocytophilum, followed by Rickettsia spp. and B. afzelii. In ticks, the most common TBP detected was Rickettsia spp., followed by Borrelia spp. and Anaplasma spp. Overall, nine different pathogens with variable prevalence were identified using species-specific primers, and the most common was Rickettsia helvetica. In three patients, there were no coincidences between the TBPs detected in whole blood and tick samples. Only in one patient was detected A. phagocytophilum in both, whole blood and tick samples. These results suggest the unequal detection of TBPs in whole blood, platelet fraction and ticks collected, from the same individual. The results justify the use of both whole blood and platelet fraction for molecular diagnosis of TBPs in patients.

Fatal Case of Imported Tick-Borne Encephalitis in South Serbia.

Tick-borne encephalitis (TBE) is vaccine-preventable neglected zoonotic neuroinvasive disease, caused by tick-borne encephalitis virus (TBEV). Many of the Central and Eastern European countries are affected by TBE, which is often poorly perceived by tourists visiting endemic territories. Here we are reporting a fatal case of imported TBE in Serbian resident who was exposed to a tick bite during a visit to Switzerland.

Mediterranean spotted fever-like illness caused by Rickettsia sibirica mongolitimonae, North Macedonia, June 2022.

Mediterranean spotted fever-like illness (MSF-like illness) is a tick-borne disease caused by Rickettsia sibirica mongolitimonae first reported in France more than 25 years ago. Until today, more than 50 cases of MSF-like illness have been reported in different regions of Europe and Africa, highlighting variable clinical manifestation. Here we report a case of MSF-like illness following a bite from a Hyalomma tick in the Skopje region of North Macedonia.

Unexpected TBEV Seropositivity in Serbian Patients Who Recovered from Viral Meningitis and Encephalitis.

The tick-borne encephalitis virus (TBEV) causes a life-threatening disease named Tick-borne encephalitis (TBE). The clinical symptoms associated with TBE range from non-specific to severe inflammation of the central nervous system and are very similar to the clinical presentation of other viral meningitis/encephalitis. In consequence, TBE is often misclassified by clinical physicians, mainly in the non-identified high-risk areas where none or only a few TBE cases have been reported. Considering this situation, we hypothesized that among persons from northern Serbia who recovered from viral meningitis or encephalitis, there would be evidence of TBEV infection. To test this hypothesis, in this observational study, we evaluated the seroreactivity against TBEV antigens in patients from northern Serbia who were hospitalized due to viral meningitis and/or viral encephalitis of unknown etiology. Three cases of seroreactivity to TBEV antigens were discovered among convalescent patients who recovered from viral meningitis and/or encephalitis and accepted to participate in the study (n = 15). The clinical and laboratory findings of these patients overlap with that of seronegative convalescent patients. Although TBE has been a notifiable disease in Serbia since 2004, there is no active TBE surveillance program for the serologic or molecular screening of TBEV infection in humans in the country. This study highlights the necessity to increase the awareness of TBE among physicians and perform active and systematic screening of TBEV antibodies among patients with viral meningitis and/or encephalitis.

Intestinal helminth infections in the golden jackal (Canis aureus L.) from Vojvodina: Hotspot area of multilocular echinococcosis in Serbia.

In the present study, 64 golden jackals were examined for intestinal helminths in three regions of Vojvodina, Serbia. Among the examined jackals 57.8% were infected with at least one parasite species. Using the intestinal scraping technique (SCT), eight species of intestinal helminths were found: Alaria alata (7.8%), Toxascaris leonina (9.4%), Toxocara canis (4.7%), Uncinaria stenocephala (20.3%), Echinococcus multilocularis (14.1%), Mesocestoides sp. (42.2%), Taenia pisiformis, and Taenia hydatigena (the overall prevalence of Taenia infection was 6.3%). To the best of our knowledge, this is the first report of T. leonina in jackals from Serbia. In comparison with the SCT results, coprological tests were less sensitive and specific for parasite identification, as only two nematode species (T. leonina and T. canis) as well as ancylostomatid and taeniid eggs were identified. The total prevalence of intestinal helminths was higher in males (71.9% males, 45% females), but the difference was not statistically significant (χ 2 = 3.76; P = 0.052). Co-infection with two species of intestinal helminths was found in 35% of the examined golden jackal individuals, three-species co-infection was demonstrated in 21.6%, whereas four-species co-infection was detected in 2.7% of the golden jackals examined. Echinococcus multilocularis has previously been recorded in jackals and foxes in Serbia, but only in Vojvodina. Our results corroborate the findings of previous studies, and indicate that the Vojvodina Province, more specifically the Srem region, is probably a high-risk area for E. multilocularis transmission to humans.

A One Health approach to study the circulation of tick-borne pathogens: A preliminary study.

Tick-borne pathogens (TBPs) have complex life cycles involving tick vectors and vertebrate hosts. However, there is limited empirical evidence on the zoonotic circulation of TBPs. In this study, we used a One Health approach to study the possible circulation of TBPs in ticks, animals and humans within a rural household in the foothills of the Fruska Gora mountain, northern Serbia. The presence of TBP DNA was assessed using microfluidic PCR (25 bacterial species, 7 parasite species, 5 bacterial genera, 3 parasite genera) in animal, human and tick samples and the presence of tick-borne encephalitis virus (TBEV) RNA was screened for using RT-qPCR on tick samples. In addition, Lyme borreliosis serology was assessed in patients sera. Rhipicephalus sanguineus and Ixodes ricinus ticks were identified on dogs and Haemaphysalis punctata was identified on house walls. Rickettsia helvetica was the most common pathogen detected in pooled R. sanguineus and I. ricinus tick samples, followed by Hepatozoon canis. None of the H. punctata tick samples tested positive for the presence of TBPs. Anaplasma phagocytophilum and Rickettsia monacensis were the most frequent pathogens detected in dogs, followed by Rickettsia felis, whereas Anaplasma bovis was the only pathogen found in one of the goats tested. None of the human blood samples collected from family members tested positive for the presence of TBPs. Although microfluidic PCR did not detect Borrelia sp. in any of the tested tick or blood samples, a family member with a history of Lyme disease was seropositive for Borrelia burgdorferi sensu lato (s.l.). We conclude that, despite the presence of TBPs in tick and vertebrate reservoirs, there is no evidence of infection with TBPs across various components of the epidemiological chain in a rural Fruska Gora household.

Tick-Borne Encephalitis Virus Seropositivity among Tick Infested Individuals in Serbia.

Tick-borne encephalitis (TBE), caused by the TBE virus (TBEV), is a life-threatening disease with clinical symptoms ranging from non-specific to severe inflammation of the central nervous system. Despite TBE is a notifiable disease in Serbia since 2004, there is no active TBE surveillance program for the serologic or molecular screening of TBEV infection in humans in the country. This prospective cohort study aimed to assess the TBEV exposure among tick-infested individuals in Serbia during the year 2020. A total of 113 individuals exposed to tick bites were recruited for the study and screened for anti-TBEV antibodies using a commercial indirect fluorescent antibody test (IFA) test. Blood samples from 50 healthy donors not exposed to tick bites were included as a control group. Most of the enrolled patients reported infestations with one tick, being I. ricinus the most frequent tick found in the participants. The TBEV seroprevalence was higher (13.27%, 15 total 113) in tick-infested individuals than in healthy donors (4%, 2 total 50), although the difference was not significant. Notably, male individuals exposed to tick bites showed five times higher relative risk (RR) of being TBEV-seropositive than healthy donors of the same gender (RR= 5.1, CI = 1.6-19; p = 0.007). None of the seropositive individuals developed clinical manifestations of TBE, but the first clinical-stage of Lyme borreliosis (i.e., erythema migrans) was detected in seven of them. Potential TBEV foci were identified in rural areas, mostly in proximity or within the Fruska Gora mountain. We conclude that the Serbian population is at high risk of TBEV exposure. Further epidemiological studies should focus on potential TBEV foci identified in this study. The implementation of active surveillance for TBEV might contribute to evaluating the potential negative impact of TBE in Serbia.

Shared Odds of Borrelia and Rabies Virus Exposure in Serbia.

Lyme borreliosis (LB) is the most common tick-borne disease in Serbia and other European countries. Rabies is a fatal zoonosis distributed worldwide and is caused by the rabies virus. Professionals at risk of rabies-including veterinarians, hunters, communal service workers, and forestry workers-overlap with some professions at a higher risk of exposure to tick bites and tick-borne pathogen infections. We hypothesized that individuals identified by the public health system as at risk of rabies virus infection, and consequently vaccinated against rabies virus, also share a higher likelihood of Borrelia exposure. To test our hypothesis, a case-control study was carried out during 2019 in Serbia to determine the seroprevalence of anti-Borrelia antibodies in two case groups (individuals at risk and vaccinated against rabies virus) and a control group (individuals without risk of rabies). Individuals vaccinated against rabies following either "pre-exposure protocol" (PrEP, n = 58) or "post-exposure protocol" (PEP, n = 42) were considered as rabies risk groups and healthy blood donors (n = 30) as the control group. The results showed higher Borrelia seroprevalence in PrEP (17.2%; 10/58) and PEP (19.0%; 8/42) groups compared with the control group (6.67%; 2/30). Furthermore, odds ratio (OR) analysis showed that risk of rabies (in either the PrEP (OR = 2.91) or PEP (OR = 3.29) groups) is associated with increased odds of being seropositive to Borrelia. However, the difference in Borrelia seroprevalence between groups was not statistically significant (Chi-square (χ²) test p > 0.05). The shared odds of LB and rabies exposure found in this study suggest that, in countries where both diseases occur, the common citizen can be at risk of both diseases when in a risky habitat. These findings are important to guide physicians in targeting high-risk groups, and diagnose LB, and to guide decision-makers in targeting control and prevention measures for both infections in risk areas.

Clinical Aspects and Detection of Emerging Rickettsial Pathogens: A "One Health" Approach Study in Serbia, 2020.

Ticks carry numerous pathogens that, if transmitted, can cause disease in susceptible humans and animals. The present study describes our approach on how to investigate clinical presentations following tick bites in humans. To this aim, the occurrence of major tick-borne pathogens (TBPs) in human blood samples (n = 85) and the ticks collected (n = 93) from the same individuals were tested using an unbiased high-throughput pathogen detection microfluidic system. The clinical symptoms were characterized in enrolled patients. In patients with suspected TBP infection, serological assays were conducted to test for the presence of antibodies against specific TBPs. A field study based on One Health tenets was further designed to identify components of a potential chain of infection resulting in Rickettsia felis infection in one of the patients. Ticks species infesting humans were identified as Ixodes ricinus, Rhipicephalus sanguineus sensu lato (s.l.), Dermacentor reticulatus, and Haemaphysalis punctata. Five patients developed local skin lesions at the site of the tick bite including erythema migrans, local non-specific reactions, and cutaneous hypersensitivity reaction. Although Borrelia burgdorferi s.l., Babesia microti, Anaplasma phagocytophilum, and Candidatus Cryptoplasma sp. DNAs were detected in tick samples, different Rickettsia species were the most common TBPs identified in the ticks. The presence of TBPs such as Rickettsia helvetica, Rickettsia monacensis, Borrelia lusitaniae, Borrelia burgdorferi, Borrelia afzelii, A. phagocytophilum, and B. microti in ticks was further confirmed by DNA sequencing. Two of the patients with local skin lesions had IgG reactive against spotted fever group rickettsiae, while IgM specific to B. afzelii, Borrelia garinii, and Borrelia spielmanii were detected in the patient with erythema migrans. Although R. felis infection was detected in one human blood sample, none of the components of the potential chain of infection considered in this study tested positive to this pathogen either using direct pathogen detection in domestic dogs or xenodiagnosis in ticks collected from domestic cats. The combination of high-throughput screening of TBPs and One Health approaches might help characterize chains of infection leading to human infection by TBPs, as well as prevalence of emerging rickettsial pathogens in the Balkan region.

Demographic and Clinical Factors Associated with Reactivity of Anti-SARS-CoV-2 Antibodies in Serbian Convalescent Plasma Donors.

Passive immunotherapy with convalescent COVID-19 plasma (CCP) is used as a therapeutic procedure in many countries, including Serbia. In this study, we analyzed the association between demographic factors, COVID-19 severity and the reactivity of anti-SARS-CoV-2 antibodies (Abs) in Serbian CCP donors. Individuals (n = 468) recovered from confirmed SARS-CoV-2 infection, and who were willing to donate their plasma for passive immunization of COVID-19 patients were enrolled in the study. Plasma samples were tested for the presence of IgG reactive to SARS-CoV-2 spike glycoprotein (S1) and nucleocapsid antigens. Individuals were characterized according to age, gender, comorbidities, COVID-19 severity, ABO blood type and RhD factor. Total of 420 candidates (420/468; 89.74%) reached the levels of anti-SARS-CoV-2 IgG that qualified them for inclusion in CCP donation program. Further statistical analysis showed that male individuals (p = 0.034), older age groups (p < 0.001), existence of hypertension (p = 0.008), and severe COVID-19 (p = 0.000) are linked with higher levels of anti-SARS-CoV-2 Abs. These findings will guide the selection of CCP donors in Serbia. Further studies need to be conducted to assess the neutralization potency and clinical efficiency of CCP collected from Serbian donors with high anti-SARS-CoV-2 IgG reactivity.

Humans infested with Ixodes ricinus are exposed to a diverse array of tick-borne pathogens in Serbia.

Tick-borne pathogens (TBPs) pose a major threat to human health in Europe and the whole northern hemisphere. Despite a high prevalence of TBPs in Ixodes ricinus ticks, knowledge on the incidence of tick-borne diseases in humans infested by this tick species is limited. This study was conducted in the year 2019 on patients who presented themselves to the Pasteur Institute Novi Sad with tick infestations. Ticks (n = 31) feeding on human (n = 30) and blood samples from the same individuals were collected by physicians and a microfluidic real-time high-throughput PCR system was used to test the genomic DNA of the samples for the presence of 27 bacterial and eight parasitic microorganisms in Serbia. Except for one Rhipicephalus sanguineus s.l. adult male tick, all ticks infesting humans were morphologically identified as I. ricinus. A high proportion of ticks (74 %, 23/31) were infected with at least one of the tested TB microorganisms, being Rickettsia helvetica (54 %, 17/31) the most common pathogen, but Borrelia afzelii (9 %, 3/31), Anaplasma phagocytophilum (6 %, 2/31), Borrelia miyamotoi (6 %, 2/31), and Francisella like-endosymbiont (6 %, 2/31), Borrelia valaisiana (3 %, 1/31), Borrelia lusitaniae (3 %, 1/31), Rickettsia felis (3 %, 1/31) and Rickettsia aeschlimannii (3 %, 1/31) were also identified. Despite the high infection rate of TBPs in ticks, only two human blood samples (6 %, 2/30) tested positive for the presence of TBPs, one patient (code H12, 67 years old female) was diagnosed with Borrelia spp. and the other patient was diagnosed (code H17, 71 years old female) with R. felis infection. The tick infesting patient H12 tested positive for B. afzelii, and R. helvetica and the tick infesting patient H17 tested positive for R. felis. Upon clinical examination, both patients were diagnosed with erythema migrans. No additional discomfort was reported by the patient and no additional pathology was observed by the physician. We concluded that humans bitten by I. ricinus in Serbia are exposed to a diverse array of TBPs with clinical impact in the Serbian cohort studied.

Molecular evaluation of chronic restrain stress in mice model of non metastatic fibrosarcoma.

Chronic stress is regarded as a significant factor in the etiology of the many diseases. Numerous methods have been developed through which the effect of chronic stress is examined. The aim of this study is to demonstrate the new experimental model for analysis of immuno-suppression induced by chronic restraint stress, through challenge with conditionally tumorigenic cell line BHK-21/C13. 20 male NMRI mice were randomly divided into 2 groups-control and experimental. Each mouse was subcutaneously inoculated with BHK-21/C13 cells. Stress in the experimental group was induced for 20 days. After the experiment, tumor masses were removed, and analyzed using histology and immunohistochemistry techniques. We found a statistically significant difference (p = 0.034) in tumor expression and tumor volumes (p = 0.0061) between groups, as well as in immunopositivity on Ki67, cytochrome C and matrix metalloproteinase 9. Absence of immune infiltrate was noticed in experimental, and the presence of inflammatory infiltrate at tumor invasion front in control group.

Significance of the red fox as a natural reservoir of intestinal zoonoses in Vojvodina, Serbia.

In the present study, 223 foxes were collected from various localities in the northern part of the Republic of Serbia (Vojvodina province) and examined for intestinal helminths. Among the examined foxes 178 (79.8%) were infected. The most frequently identified parasites were Mesocestoides spp. (49.3%) and Toxascaris leonina (36.3%). The parasite with the lowest prevalence was Pterygodermatites affinis (0.9%), and this is the first confirmed finding in Serbia. The other recovered species were Alaria alata (25.6%), Taenia spp. (6.3%), Echinococcus multilocularis (13%), Toxocara canis (16.6%) and Uncinaria stenocephala (14.8%). The highest number of foxes infected with E. multilocularis were in the Srem area. The results of this study indicate the presence of helminth species in red foxes in Vojvodina which may also infect humans.

Mysterious path of Borrelia spielmanii: spreading without morphological alteration of collagen type I and IV.

The majority of suggested mechanisms of Borrelia spreading inside erythema migrans (EM) are developed from in vitro studies and animal models. This report is the first to describe pathomorphological substrate of EM caused by Borrelia spielmanii in humans, addressing the hypothesis of enhanced Borrelia penetration through extracellular matrix. In the process of ruling out of atypical Masters' disease, we conducted a punch biopsy of suspected EM and a two-tier serology testing for Lyme borreliosis, where we registered antibodies against B. spielmanii. Skin biopsy showed CD4+ and CD8+ lymphocyte involvement and high activity of matrix metalloproteinase 9. No alterations were detected in distribution and morphology of collagen type I and IV. Therefore, it is suggested that other mechanisms should be considered as major contributing factors to local spreading of B.spielmanii.

Comparison of Conventional and Molecular Diagnostic Techniques for Detection of Blastocystis sp. in Pig Faeces.

BACKGROUND: Blastocystis is a common protist colonizing the gastrointestinal tract of humans and various animals. Pigs have been suggested to be a reservoir for human Blastocystis infections because of high prevalence of the parasite in these animals and the presence of zoonotic subtypes. Nevertheless, epidemiological data is often misinterpreted due to the lack of standard diagnostic procedures. This study aimed to compare the sensitivity of different diagnostic techniques in detection of Blastocystis sp. in pigs. METHODS: Overall, 48 individual faecal samples were collected from pigs reared in an intensive farming system (Autonomous Province of Vojvodina, Serbia) and were tested by microscopic examination of direct wet mount, in vitro cultivation in modified Jones' medium and conventional PCR for rRNA gene. RESULTS: Xenic in vitro cultivation in Jones' medium showed higher sensitivity than direct wet mount when we compared it with PCR. Namely, the estimated sensitivity of direct wet mount was 46.15%, while the sensitivity of in vitro cultivation was 84.62%. CONCLUSION: Low sensitivity of conventional parasitological compared to molecular methods is proven. Thus, reports on prevalence that rely solely on microscopy of faecal samples (unprocessed or concentrated) are probably underestimating the true prevalence of the parasite.