The distribution and prevalence of zoonotic pathogens infecting ixodid ticks in Western Europe have been extensively examined. However, data on ticks and tick-borne pathogens in Eastern Europe, particularly Ukraine are scarce. The objective of the current study was, therefore, to investigate the prevalence of Anaplasma phagocytophilum, Anaplasmataceae, Rickettsia spp., Babesia spp., Bartonella spp., and Borrelia burgdorferi sensu lato in engorged and questing ixodid ticks collected from five administrative regions (oblasts) of Ukraine, namely Chernivtsi, Khmelnytskyi, Kyiv, Ternopil, and Vinnytsia. The ticks were collected from both wild and domestic animals and from vegetation. Of 524 ixodid ticks collected, 3, 99, and 422 ticks were identified as Ixodes hexagonus, Ixodes ricinus, and Dermacentor reticulatus, respectively. DNA samples individually extracted from 168 questing and 354 engorged adult ticks were subjected to pathogen-specific PCR analyses. The mean prevalence in I. ricinus and D. reticulatus were, respectively: 10 % (10/97) and 3 % (12/422) for A. phagocytophilum; 69 % (67/ 97) and 52 % (220/422) for members of the Anaplasmataceae family; 25 % (24/97) and 28 % (117/422) for Rickettsia spp.; 3 % (3/97) and 1 % (6/422) for Babesia spp.; and 9 % (9/97) and 5 % (20/422) for Bartonella spp. Overall, between the five cities, there was no significant difference in the prevalence of any of the pathogens for the respective ticks (p > 0.05). The prevalence of B. burgdorferi s. l. in the questing and engorged I. ricinus varied from 0 to 27 % and 14-44%, respectively, with no statistical significance identified between the five cities (p > 0.05). In addition to reporting the updated data for Kyiv and Ternopil, this study is the first to provide the prevalences of the tick-borne pathogens for Chernivtsi, Khmelnytskyi, and Vinnytsia. This investigation is also the first to detect Neoehrlichia mikurensis in ixodid ticks from Ukraine. These new data will be useful for medical and veterinary practitioners as well as public health officials when diagnosing infections and when implementing measures to combat tick-borne diseases in Ukraine.

Canine babesiosis is a tick-borne disease caused by parasites of the genus Babesia. Tumour necrosis factor alpha (TNF-α) is a cytokine that plays a role in the pathogenesis of canine babesiosis. In this study, the authors determined the concentration of serum TNF-α in 11 dogs infected with Babesia canis and calculated Spearman’s rank correlations between the concentration of TNF-α and blood pressure, and between TNF-α and indices of renal damage such as: fractional excretion of sodium (FE(Na⁺)), urinary creatinine to serum creatinine ratio (UCr/SCr), renal failure index (RFI), urine specific gravity (USG) and urinary protein to urinary creatinine ratio (UPC). The results demonstrated statistically significant strong negative correlations between TNF-α and systolic arterial pressure (r = −0.7246), diastolic arterial pressure (r = −0.6642) and mean arterial pressure (r = −0.7151). Serum TNF-α concentration was also statistically significantly correlated with FE(Na⁺) (r = 0.7056), UCr/SCr (r = −0.8199), USG (r = −0.8075) and duration of the disease (r = 0.6767). The results of this study show there is an increase of serum TNF-α concentration during canine babesiosis, and the increased TNF-α concentration has an influence on the development of hypotension and renal failure in canine babesiosis. This probably results from the fact that TNF-α is involved in the production of nitric oxide and induction of vasodilation and hypotension, which may cause renal ischaemia and hypoxia, and finally acute tubular necrosis and renal failure.

The effect of Toxocara larval antigens on cytokine secretion by mouse splenocytes was studied in vitro. Recombinant mucins were produced in Pichia pastoris yeast, and Toxocara excretory-secretory (TES) antigens were collected from in vitro culture of L2 larvae. Tc-MUC-2, Tc-MUC-3, Tc-MUC-4, and Tc-MUC-5 were expressed as glycoproteins and were specifically recognized by Toxocara canis-infected dog serum antibodies. Mouse splenocytes stimulated with recombinant mucins produced IL-5, IL-6, and TGF-β. Cell stimulation with whole TES products was more effective and resulted in secretion of IL-4, IL-5, IL-6, IL-10, and TGF-β and downregulation of TNF-α production. IFN-γ and IL-17 secretion was noted only after ConA treatment. Cells originating from infected animals produced significantly smaller amounts of these two cytokines compared to control cells, which suggests that Th1 and Th17 response in infected mice is strongly inhibited. However, splenocyte stimulation with both TES and ConA upregulated the production of IFN-γ and IL-17. This shows that TES antigens have strong immunomodulatory properties and are able to induce a broad range of effects on murine immune cells.

Twenty-nine Lactobacillus plantarum strains isolated from different types of Polish regional cheeses (Oscypek and Korycinski) were assessed for selected probiotic properties and anti-staphylococcal activity. Most of the tested L. plantarum strains were considered safe. Whole bacterial cultures (WBC) and cell-free supernatants (CFSs) of L. plantarum strains inhibited growth of Staphylococcus aureus (average inhibition growth zones were 2.8 mm ± 1.2 and 2.8 mm ± 1.1 respectively). Moreover, almost all neutralized, catalase-treated cell-free supernatants (CFN) of L. plantarum cultures also exhibited slight anti-staphylococcal activity in vitro. The most promising strains Os4 and Kor14 were selected for further study. Both strains were able to survive during digestive gastro-intestinal passage model. Live cells of L. plantarum Os4 and Kor14 caused the strongest inhibition of S. aureus adhesion to Caco-2 cells comparing with CFN and heat-killed bacterial cells. S. aureus and L. plantarum (Os4 or Kor14) co-cultured in skim milk resulted in growth inhibition of S. aureus in both 8 °C and 37 °C incubation temperatures. Observed abilities, demonstrated for L. plantarum Os4 and Kor14, confirms that these strains could be used in the food industry as protective cultures.

In the current study, the probiotic activity of ten Lactobacillus (Lb.) strains, previously isolated from the traditional Polish fermented vegetable, was characterized. Strains were assessed for adhesion to human intestinal epithelial Caco-2 cells and regulation of selected cytokine production (IL-1β, IL-6, IL-10, IL-23, and TNF-α) by THP-1 macrophages. The effect of tested strains on Caco-2 cell apoptosis was also investigated using a caspase-3 assay. Adhesion capacity was strain-dependent (1.29–8.24% of initial population). Highest adhesion was observed for Lb. brevis O24. All Lactobacillus strains investigated in this study stimulated two- to threefold increase in TNF-α, IL-1β, and IL-6 production, compared to the control. Additionally, selected strains of Lactobacillus caused a significant decrease of pro-inflammatory cytokine production by lipopolysaccharide (LPS)-stimulated THP-1 cells. Almost all Lactobacillus investigated in this study are potent stimulators of IL-10 production. All tested Lactobacillus cells slightly increased the caspase-3 activity in Caco-2 cells. Lb. casei O18 was the most inducing strain. The tested strains had no effect on staurosporine (STS)-induced caspase-3 activity. According to these results, the most promising strains are Lb. plantarum O20, two strains Lb. brevis O22 and O24, and Lb. rhamnosus K3. These newly identified lactobacilli hold promise for use as probiotics in functional food applications.

The effect of Toxocara canis antigens on cytokine production by human THP-1 macrophages was studied in vitro. Toxocara Excretory–Secretory products (TES) and recombinant mucins (Tc-MUC-2, Tc-MUC-3, Tc-MUC-4, and Tc-MUC-5) as well as deglycosylated forms of these antigens were used in the study. TES products stimulated macrophages to produce the innate proinflammatory IL-1β, IL-6, and TNF-α cytokines regardless of the presence of glycans. Recombinant mucins induced glycan-dependent cytokine production. Sugar moieties led to at least 3-fold higher production of regulatory IL-10 as well as proinflammatory cytokines. The presence of glycans on mucins also affected the downstream signalling pathways in stimulated cells. The most prominent difference was noted in AKT and AMPK kinase activation. AKT phosphorylation was observed in cells stimulated with glycosylated mucins, whereas treatment with deglycosylated antigens led to AMPK phosphorylation. MAP kinase family members such as JNK and p38 and c-Jun transcription factor were phosphorylated in both cases what suggests that toll-like receptor signalling may be involved in mucin-treated macrophages. This pathway is however modified by other signalling molecules as only mucins containing intact sugars significantly induced the production of cytokines.