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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.

Mediterranean spotted fever-like illness (MSF-like illness) is a tick-borne disease caused by

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.

Objective. To test if the direct immunofluorescence can be used for the detection of Borrelia afzelii in brain tissue during the standardization of the animal model of neuroborreliosis in NMRI mice. Methods. The study was performed on 15 mice of NMRI strain. All mice were subcutaneously inoculated with 100 ml of BSK-H medium containing the local isolate of Borrelia afzelii. Animals were sacrificed after inoculation at III (n = 4), IV (n = 6) and V (n = 5) weeks, by cervical dislocation. In the sampled brains of mice, the presence of Borrelia was detected by direct immunofluorescence (DIF) and chain polymerization reaction (PCR). Results. The first brain tested positive for Borrelia three weeks after the inoculation. The bacteria were detected in 1 out of 4 brains (25%). After that, there was a growth in the percentage of positive results. The data showed that 3 out of 6 brains (50%) were found positive on Borrelia presence by the end of the fourth week. Whereas, in 3 out of 5 brains (60%) Borrelia was detected five weeks following the inoculation. Conclusion. According to the preliminary results, direct immunofluorescence appeared to be a practical, low budget method for following the kinetics of neuro-infection. NMRI mice could be considered as an adequate animal model for neuroborreliosis. Thus, more research is needed on the topics of infection kinetics for the period after fifth week post inoculation, as well as sensitivity and specificity of direct immunofluorescence.

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.

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.

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.

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.

Summary As part of routine monitoring of foxes (Vulpes vulpes) and jackals (Canis aureus) on the territory of Vojvodina province (northern Serbia), an analysis of respiratory and cardiopulmonary parasitic nematodes was conducted. Both host species harbored Eucoleus aerophilus, E. boehmi and Crenosoma vulpis, whereas Angiostrongylus vasorum was found only in foxes. A high prevalence of infection (72.6 %) was noted for E. aerophilus in foxes. The remaining parasite species occurred less frequently in both host species. In all species where it could be quantified, a high degree of parasite aggregation within host individuals was noted. Single species infections were most common, whereas two and three species infections occurred less frequently in both host species. The distribution of abundance of E. aerophilus was affected by host sex, with abundances higher in male foxes. Sampling site and year influenced abundance variation in E. boehmi.

The majority of suggested mechanisms of

International audience; 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.

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.