Prevalence of Bacterial and Protozoan Pathogens in Ticks Collected from Birds in the Republic of Moldova.

Epidemiological knowledge on pathogens in ticks feeding on birds in Moldova is scarce. To reduce this gap of information, a total of 640 migrating and native birds of 40 species were caught from 2012 to 2015 and examined for the presence of ticks in the Republic of Moldova. Altogether, 262 ticks belonging to five tick species (Ixodes ricunus n = 245, Ixodes frontalis n = 12, Haemaphysalis punctata n = 2, Hyalomma marginatum n = 2 (only males), Dermacentor marginatus n = 1) were collected from 93 birds. Of these ticks, 250 (96%) were at the stage of a nymph and 9 at the stage of a larva (3%). One imago of I. frontalis and two imagoes of Hy. marginatum were found. Generally, ticks infested 14.1% of the assessed birds belonging to 12 species. DNA was extracted from individual ticks with subsequent PCR targeting Rickettsia spp., Borrelia spp. in general, as well as relapsing fever-associated Borrelia spp., in particular, Anaplasma phagocytophilum, Neoehrlichia mikurensis, Babesia spp. and Coxiella burnetii. The bird species Turdus merula showed the heaviest infestation with ticks and the highest incidence of infected ticks. Altogether, 32.8% of the assessed ticks (n = 86) were positive for one of the pathogens. DNA of Borrelia spp. was found in 15.2% (40/262) of the investigated ticks; in 7.6% of ticks (20/262), DNA of rickettsiae was detected; 6.9% (18/262) of the ticks were positive for A. phagocytophilum DNA; in 1.5% of the ticks (4/262), DNA of Neoehrlichia mikurensis was detected, followed by 1.5% (4/262) Babesia microti and 1.5% (4/262) Borrelia miyamotoi. Within the B. burgdorferi complex, B. garinii (n = 36) was largely predominant, followed by B. valaisiana (n = 2) and B. lusitaniae (n = 2). Among the detected Rickettsia spp., R. monacensis (n = 16), R. helvetica (n = 2) and R. slovaca (n = 1) were identified. In conclusion, the study provided some new information on the prevalence of ticks on birds in Moldova, as well as the presence of DNA of pathogens in the ticks. By doing so, it provided an additional piece in the puzzle of the global epidemiology of tick-transmitted infectious diseases from a geographic side from where respective surveillance data are scarce.

Biological Evaluation of a Series of Amine-Containing Mixed-Ligand Copper(II) Coordination Compounds with 2-(2-hydroxybenzylidene)-N-(prop-2-en-1-yl)hydrazinecarbothioamide.

Five compounds 2-(2-hydroxybenzylidene)-N-(prop-2-en-1-yl)hydrazinecarbothioamide (H2L), bis[µ2-2-({2-[(prop-2-en-1-yl)carbamothioyl]hydrazinylidene}methyl)phenolato-S,N,O:O]diaquadicopper(II) nitrate (1), bis[µ2-2-({2-[(prop-2-en-1-yl)carbamothioyl]hydrazinylidene}methyl)phenolato-S,N,O:O]diimidazoldicopper(II) nitrate (2), bis[µ2-2-({2-[(prop-2-en-1-yl)carbamothioyl]-hydrazinylidene}methyl)phenolato-S,N,O:O]bis-(3,5-dibromopyridine)dicopper(II) nitrate (3), bis[µ2-2-({2-[(prop-2-en-1-yl)carbamothioyl]-hydrazinylidene}methyl)phenolato-S,N,O:O]bis(4-methylpyridine)dicopper(II) nitrate hexahydrate (4) were synthesized. The antiproliferative properties of these compounds toward cancer cell lines RD, HeLa, and normal cell line MDCK have been investigated. The tested complexes surpass Doxorubicin (DOXO) in the efficiency of anticancer activity as their IC50 values toward cancer cells are lower than the corresponding values of DOXO and the selectivity indexes exceed the corresponding SI value of DOXO. The tested compounds demonstrated a high antioxidant effect against ABTS•+ radical cations as well as low toxicity on Daphnia magna.

Screening of biological properties of MoV2O2S2- and MoV2O4-based coordination complexes: Investigation of antibacterial, antifungal, antioxidative and antitumoral activities versus growing of Spirulina platensis biomass.

This paper deals with the biological potential of coordination compounds based on binuclear core [MoV2O2E2]2+ (E = O or S) coordinated with commercially available ligands such as oxalates (Ox2-), L-cysteine (L-cys2-), L-histidine (L-his-), Iminodiacetate (IDA2-), Nitrilotriacetate (HNTA2- or NTA3-) or ethylenediamine tetraacetate (EDTA4-) by means of various in vitro assays in a screening approach. Results suggest that the obtained complexes show weak antibacterial and antifungal properties while not being cytotoxic on cancerous and mammalian cells. In contrast, [Mo2O2E2(L-cys)2]2- complexes stand out as powerful antioxidant, whereas [Mo2O2E2(EDTA)]2- associating tetraphenylphosphonium counter-cations display strong antibiotic activity. Finally, some complexes have evidenced a positive activity towards the growing of spirulina platensis together with a modification of the proportions of biological components inside the cells. These findings reveal promising bioactivity of the bridged binuclear Mo(+V) cores inside complexes and encourage further research for new highly active yet non-toxic molecules for biological and biomedical applications.

Abamectin Efficacy on the Potato Cyst Nematode Globodera pallida.

The potato cyst nematode Globodera pallida is a major pest of the potato crop. Abamectin is a biological pesticide showing high nematicide activity, but its efficacy to control G. pallida has not been investigated to date. In this study, combination of different abamectin concentrations ranging from 1.125 to 36 µg/mL x exposure times from 24 to 384 h were tested on the nematode in a hatching test. Abamectin induced mortality with LD50 value in the range of 13.23 (after 24 h) to 2.90 µg/mL (after 384 h). A glasshouse experiment was also performed in pots filled with soil infected with G. pallida in the presence of sprouted potato tubers cultivar "Spunta". Abamectin at 4.5, 9.0, 18.0 and 36.0 µg/mL was used in comparison with nematicide fosthiazate. The doses of 18 and 36 µg/mL significantly reduced number of eggs, juveniles, cyst/g soil and reproduction rate in comparison to both untreated control and fosthiazate treatment. Soil applications of abamectin provided significant G. pallida control with LD50 and LD99.9 of 14.4 and 131.3 µg/mL, respectively. These results indicate the efficacy of abamectin to control G. pallida on potato crops and its potential use in organic agriculture or in an integrated pest management program.

Age-dependent effect between MARCO and TLR4 on PMMA particle phagocytosis by macrophages.

Progressive generation of total joint implant-derived wear particles is one of the major risk factors in development of peri-prosthetic osteolysis especially in the aging society. It is commonly accepted that macrophages predominantly drive the inflammatory response to wear debris particles. Among various surface receptors that activate the macrophages to phagocytize particles, it is believed that the Toll-like receptor 4 (TLR4) and the scavenger macrophage receptor with collagenous structure (MARCO) play key roles in recognition of wear debris particles. However, a strong body of evidence indicates an age-dependent diminished function of human TLRs. Thus, we hypothesized that the MARCO receptor may be more engaged than TLRs in the phagocytosis of wear debris particles which in turn up-regulate production of pro-inflammatory cytokines from aged macrophages. We demonstrated that peritoneal macrophages isolated from aged mice show elevated expression of MARCO receptor compared to that from young mice. In contrast the expression of TLR4 was significantly decreased on the surface of aged macrophages. Furthermore, using anti-MARCO and anti-TLR4 neutralizing mAbs, we demonstrated the age-dependent pathogenic role of MARCO, but not TLR4, receptor in promoting poly-methyl methacrylate (PMMA) bone cement particles phagocytosis by macrophages leading to the release of pro-inflammatory cytokines migration inhibitory factor and tumour necrosis factor in vitro. These data also suggest that the approach to neutralize MARCO may lead to the development of therapeutic regimen for the prevention of particle-induced osteolysis in aged patients.

Change of strategy or always overlooked? Third recent record of the beardless tadpole goby Benthophiloides brauneri from an inland location far from the coast.

The present study documents three specimens of Benthophiloides brauneri from the River Dniester. It is one of the rarest freshwater fishes in Europe, living along the western coast of the Black and Asov Seas, but these new records are 200 river km from the coast. We discuss whether the species has recently started to establish inland populations or inland specimens have previously been overlooked.

Comparison of tick-borne microorganism communities in Ixodes spp. of the Ixodes ricinus species complex at distinct geographical regions.

Characterizing the tick-borne microorganism communities of Ixodes ricinus (sheep tick) and Ixodes persulcatus (taiga tick) from the I. ricinus species complex in distinct geographical regions of Eastern Europe and European Russia, we demonstrated differences between the two ticks. Taiga ticks were more frequently mono- and co-infected than sheep ticks: 24.4 % (45/184 tested ticks) versus 17.5 % (52/297) and 4.3 % (8/184) versus 3.4 % (10/297), respectively. Ginsberg co-infection index values were significant at the various sites. Diversity of the tick-borne microorganism communities was estimated by the Shannon index, reaching values of 1.71 ± 0.46 and 1.20 ± 0.15 at the sheep-tick and the taiga-tick harbored sites, respectively. Richness of the tick-borne microorganism community in the sheep tick collection sites was about twice the value of the taiga tick collection sites. Future investigations are warranted to further characterize the peculiarities of the tick-borne microorganism communities among the ticks of the Ixodes ricinus complex.

Annotated checklist of the mosquitoes of the Republic of Moldova.

The mosquito fauna of the Republic of Moldova is poorly known. In an effort to understand the Culicidae fauna better, mosquito collections have been conducted between early April and middle November from 2008 to 2012. A total of 10,923 larval specimens and 8,246 adults were collected from 20 regions of Moldova. Altogether 36 species have been recorded during the recent study, bringing the total Moldovan mosquito fauna to 40 species in 9 genera and 11 subgenera. New state records include the following 7 species: Anopheles pseudopictus, An. melanoon, Aedes geminus, Culex torrentium, Culiseta longiareolata, Coquillettidia buxtoni, and Uranotaenia unguiculata.

Molecular detection of tick-borne pathogens in Ixodes ricinus from Moldova collected in 1960.

This study is the first report about the prevalence of tick-borne pathogens, as well as their (co-)infection rates, in the museum-archived I. ricinus female ticks collected in Moldova in 1960. A total of 16.7% (21/126) ticks was mono-infected. Borrelia burgdorferi sensu stricto was revealed as the most abundant species (4.8%) followed by B. garinii (1.6%), B. afzelii (0.8%), B. valaisiana (0.8%), and B. lusitaniae (0.8%). DNA of Rickettsia helvetica (2.4%), R. monacensis (2.4%), Anaplasma phagocytophilum (2.4%), 'Candidatus Neoehrlichia mikurensis' (0.8%), and Babesia microti (0.8%) were also detected, indicating the occurrence of these emerging tick-borne microorganisms in Moldova since 1960 at least. In this study, we detected a co-infection (0.8%; 1/126 tested ticks) between B. microti and R. helvetica. Additional investigations are warranted to further characterize a historical snapshot of the distribution of tick-borne pathogens in Europe.

Detection of Babesia Sp. EU1 and members of spotted fever group rickettsiae in ticks collected from migratory birds at Curonian Spit, North-Western Russia.

To reveal the prevalence of spotted fever group (SFG) rickettsiae and Babesia sp. in Ixodes ricinus (L.) ticks from migratory birds, 236 specimens represented 8 species of Passeriformes and were collected at Curonian Spit in Kaliningrad enclave of North-Western Russia. The ticks (total 126) being detached from four bird species, Turdus philomelos, Fringilla coelebs, Parus major, and Sturnus vulgaris, were investigated by PCR using the primers Rp CS.877p/Rp CS.1258n for the detection of Rickettsia and BJ1/BN2 for Babesia spp. Babesia spp. were detected in 2 of 126 (1.6%) ticks. The partial sequence of 18S rDNA had 100% similarity to human pathogenic Babesia sp. EU1. The SFG rickettsiae were detected in 19 of 126 (15.1%) ticks collected from the above-mentioned bird species. BLAST analysis of SFG rickettsia gltA assigned sequences to human pathogenic Rickettsia helvetica (10.3%), Rickettsia monacensis (3.9%), and Rickettsia japonica (0.8%) with 98%-100% sequence similarity. The SFG rickettsiae and Babesia sp. EU1 in ticks collected from the passerines in Russia were detected for the first time. The survey indicates that migratory birds may become a reservoir for Babesia spp. and SFG rickettsiae. Future investigations need to characterize the role of birds in the epidemiology of these human pathogens in the region.

First report of Anaplasma phagocytophilum and its co-infections with Borrelia burgdorferi sensu lato in Ixodes ricinus ticks (Acari: Ixodidae) from Republic of Moldova.

We examined 198 questing Ixodes ricinus ticks collected in Chisinau City, Republic of Moldova by PCR assays for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and co-infection of both pathogens, which were detected in 9%, 25.2% and 2.5% of tested ticks, respectively. B. burgdorferi s.l. genotyping revealed the presence of five genospecies with dominance of B. garinii. Our preliminary study provides evidence about occurrence of both pathogens in this populated area, which represent a potential health risk for inhabitants.