Antiparasitic Activity of Enterocin M and Durancin-like from Beneficial Enterococci in Mice Experimentally Infected with Trichinella spiralis.

Beneficial/probiotic strains protect the host from pathogens by competitive displacement and production of antibacterial substances, i.e., bacteriocins. The antiparasitic potential of bacteriocins/enterocins and their producing strains in experimental murine trichinellosis were tested as a new therapeutic strategy. Enterocin M and Durancin-like and their producers Enterococcus faecium CCM8558 and Enterococcus durans ED26E/7 were administered daily to mice that were challenged with Trichinella spiralis. Our study confirmed the antiparasitic effect of enterocins/enterococci, which reduced the number of adults in the intestine (Enterocin M-43.8%, E. faecium CCM8558-54.5%, Durancin-like-16.4%, E. durans ED26E/7-35.7%), suppressed the Trichinella reproductive capacity ex vivo (Enterocin M-61%, E. faecium CCM8558-74%, Durancin-like-38%, E. durans ED26E/7-66%), and reduced the number of muscle larvae (Enterocin M-39.6%, E. faecium CCM8558-55.7%, Durancin-like-15%, E. durans ED26E/7-36.3%). The direct effect of enterocins on Trichinella fecundity was documented by an in vitro test in which Durancin-like showed a comparable reducing effect to Enterocin M (40-60%) in contrast to the ex vivo test. The reducing activity of T.spiralis infection induced by Enterocin M was comparable to its strain E. faecium CCM8558; Durancin-like showed lower antiparasitic activity than its producer E. durans ED26E/7.

Modulatory Effect of Beneficial Enterococci and Their Enterocins on the Blood Phagocytes in Murine Experimental Trichinellosis.

Bacteriocins (enterocins) represent a new therapeutic strategy in various intestinal and non-intestinal infections. In antiparasitic defence, an oxidative inflammation of phagocytes is effective in destroying new-born Trichinella spiralis larvae. The strains Enterococcus faecium CCM8558 and E. durans ED26E/7 and their enterocins, enterocin M and a durancin-like enterocin, respectively, were administered daily, and mice were then infected with T. spiralis larvae on the seventh day of treatment. Phagotest and Bursttest kits were used to detect the phagocytosis and respiratory burst in blood leukocytes. T. spiralis infection inhibited phagocytosis from day 11 post-infection (dpi) during the migration of new-born larvae into the muscles. E. faecium CCM8558, E. durans ED26E/7, and the durancin-like enterocin increased phagocytic activity from day 11 dpi. Both strains and their enterocins (enterocin M and durancin-like) stimulated the ingestion capability of phagocytes from 18 to 32 dpi. Enterococci/enterocins therapy prevented a reduction in cells with respiratory burst caused by T. spiralis infection from 11 dpi. The enzymatic activity of phagocytes was stimulated on 18 and 25 dpi, particularly by E. faecium CCM8558 and enterocin M. Enterocin M and the durancin-like enterocin were as effective in stimulating phagocytosis as the bacterial strains that produce them. The stimulation of phagocytosis could contribute to decreased larval migration and reduced parasite burden in the host.

Dirofilaria immitis conquering the regions in Slovakia previously endemic for D. repens.

The present study was focused on the current state of Dirofilaria species distribution in the territory of the Slovak Danubian Lowland, a region previously identified endemic for Dirofilaria repens. For the research, blood samples of 330 dogs tested positive for dirofilariosis using concentration tests or "rapid heartworm tests" were sent by private veterinary practitioners for further DNA analyses and species determination. The results revealed an unquestionable change in the pattern of Dirofilaria species distribution with Dirofilaria immitis, diagnosed as the agent of mono- and co-infections with D. repens, responsible for 60.00% of all cases. The results showed that in the course of the last 5 years, D. immitis has spread significantly in Slovakia and has become the dominant causal agent of dirofilariosis in the former endemic areas of D. repens distribution, which increases infection risk for both dogs and humans.

A Study of Oribatid Mites as Potential Intermediate Hosts of Anoplocephalid Tapeworms of Tatra chamois and Tatra marmots from the Tatra Mountains, Central Europe, and Report of a New Intermediate Host for Andrya cuniculi, the Parasite of Leporidae.

Tatra chamois (Rupicapra rupicapra tatrica (Blahout 1972)) and Tatra marmot (Marmota marmota latirostris (Kratochvíl 1961)) are significant endemic subspecies of the subalpine and alpine ranges of the Tatra Mountains in Central Europe. In four studied localities in the range of their typical biotopes in Slovakia and Poland, we investigated intestinal parasites of Tatra chamois and Tatra marmots, with an emphasis on anoplocephalid tapeworms. We also studied the occurrence, species diversity, and abundance of oribatid mites as intermediate hosts thereof, and the prevalence of cysticercoid larval stages of anoplocephalid tapeworms in collected oribatids using morphological and molecular methods. Coprological analyses revealed the average positivity of Moniezia spp. in chamois faeces at 23.5% and Ctenotaenia marmotae in marmot samples at 71.1%, with significant differences between the localities under study. Morphological analyses determined the presence of cysticercoids in five oribatid species: Ceratozetes gracilis, Edwardzetes edwardsi, Scheloribates laevigatus, Trichoribates novus, and Tectocepheus velatus sarekensis. This is the first record of T. v. sarekensis as an intermediate host of anoplocephalid tapeworms, as well as the first report of Andrya cuniculi occurrence in the territory of the Tatra Mountains, confirmed also by molecular methods.

Systematic Review and Modelling of Age-Dependent Prevalence of Toxoplasma gondii in Livestock, Wildlife and Felids in Europe.

Toxoplasma gondii is a zoonotic parasite of importance to both human and animal health. The parasite has various transmission routes, and the meat of infected animals appears to be a major source of human infections in Europe. We aimed to estimate T. gondii prevalence in a selection of animal host species. A systematic literature review resulting in 226 eligible publications was carried out, and serological data were analyzed using an age-dependent Bayesian hierarchical model to obtain estimates for the regional T. gondii seroprevalence in livestock, wildlife, and felids. Prevalence estimates varied between species, regions, indoor/outdoor rearing, and types of detection methods applied. The lowest estimated seroprevalence was observed for indoor-kept lagomorphs at 4.8% (95% CI: 1.8-7.5%) and the highest for outdoor-kept sheep at 63.3% (95% CI: 53.0-79.3%). Overall, T. gondii seroprevalence estimates were highest within Eastern Europe, whilst being lowest in Northern Europe. Prevalence data based on direct detection methods were scarce and were not modelled but rather directly summarized by species. The outcomes of the meta-analysis can be used to extrapolate data to areas with a lack of data and provide valuable inputs for future source attribution approaches aiming to estimate the relative contribution of different sources of T. gondii human infection.

Pilot research on gastrointestinal parasites of the Tatra chamois (Rupicapra rupicapra tatrica).

INTRODUCTION AND OBJECTIVE: The Tatra chamois (Rupicapra rupicapra tatrica) is a significant representative of the High Tatra Mountains endemic fauna species. In terms of health hazards for these animals, an important role is played by parasitic infections that can lead to a significant depletion of the entire population. OBJECTIVE: The aim of the study was to describe the occurrence of gastrointestinal parasites of Tatra chamois in the current environmental and climatic conditions of the High Tatra Mountains. MATERIAL AND METHODS: During the pilot project in 2014 - 2017, a total of 494 chamois faecal samples were collected from the Slovak High Tatra and 114 samples in the Polish part of the mountain and examined using standard coprological methods. RESULTS: The results revealed that the overall positivity for gastrointestinal parasites in chamois of the Slovak High Tatra reached 74.7%. Most frequent were protozoa - Eimeria spp. (42.7%), helminths were represented by Moniezia spp. tapeworms (23.5%), eggs of GIS-strongylids (7.1%), and sporadically Capillaria spp. (1.4%). The chamois from the Polish Tatra Mts. were infected with Eimeria spp. (43.9%), GIT-strongylids (9.6%), and Moniezia spp. (6.1%). Parasitic infection was determined in 59.6 % of faecal samples from the Polish part of the mountains. Statistical analyses revealed a significant difference in Moniezia spp. occurrence in different Slovak Tatra Mts. Regions, as well as between Slovak and Polish Tatra Mts. CONCLUSIONS: Initial research on the gastrointestinal parasites of the Tatra chamois revealed one indisputable finding - a relatively high prevalence of the genus Moniezia, which is closely linked to the climate and microclimate conditions of the mountains. Further intensive research on parasite composition and distribution in Tatra chamois is needed in broader temporal, ecological, and zoological contexts.

Modulation of lymphocyte subpopulations in the small intestine of mice treated with probiotic bacterial strains and infected with Trichinella spiralis.

AIMS: To study the local intestinal lymphocyte immunity in mice with trichinellosis affected by probiotic bacteria. METHODS AND RESULTS: Enterococcus faecium CCM8558, Enterococcus durans ED26E/7, Limosilactobacillus fermentum CCM7421 and Lactiplantibacillus plantarum 17 L/1 were administered daily (109  CFU ml-1 ) and mice were infected with Trichinella spiralis (400 larvae) on the 7th day of treatment. T. spiralis infection significantly inhibited lymphocyte subpopulations from 5 to 25 days postinfection (dpi). L. fermentum CCM7421 and L. plantarum 17 L/1 restored the CD4+ T cell numbers in the epithelium and lamina propria at the control level from 11 dpi. All strains stimulated the CD8+ T cells numbers in infected mice, which were restored in the lamina propria on 11 dpi and in the epithelium only on 32 dpi. B cells (CD19+ ) inhibition after T. spiralis infection was not affected by treatment till 25 dpi. CONCLUSIONS: The strain-specific immunomodulatory effect of tested bacteria was confirmed. L. fermentum CCM7421 and L. plantarum 17 L/1 showed the greatest immunomodulatory potential on CD4+ and CD8+ T lymphocytes in trichinellosis. E. faecium CCM8558 and E. durans ED26E/7 activated only CD8+ T cells in the lamina propria. SIGNIFICANCE AND IMPACT OF THE STUDY: Positive modulation of the gut lymphocyte immunity in T. spiralis infection with bacterial strains showed their beneficial effect with the host's antiparasitic defence.

Thelazia callipaeda in Slovakia - From sporadic cases to endemic areas.

Spirurid nematode Thelazia callipaeda, transmitted by the fruit fly Phortica variegata, is a causative agent of an ocular parasitic disease called also canine thelaziosis. Dogs, cats, and wild canids are considered the primary definitive hosts for the parasite, but humans may also serve as aberrant definitive hosts. For long decades the geographic range of T. callipaeda was strictly limited to the territory of Asia, but after the year 2000, the parasite began to spread rapidly through Europe. The first autochthonous infections of dogs and foxes in Slovakia were recorded in 2016. In the present study, the results of a whole-area surveillance for canine thelaziosis are reported. Altogether, 142 cases of infection caused by T. callipaeda were diagnosed by veterinarians in dogs between 2016 and the first quarter of 2021, and two cases of feline thelaziosis were recorded. The majority of the dogs showed mild ocular signs manifested by conjunctivitis; 8.5% of them suffered from more serious mucopurulent discharge, and in two dogs corneal ulceration was recorded. The screening revealed increasing trends in the occurrence of canine thelaziosis from both a temporal and spatial point of view and unambiguously confirms the endemic status of T. callipaeda in Slovakia with the prospect of its further expansion.

Mosquito surveillance of West Nile and Usutu viruses in four territorial units of Slovakia and description of a confirmed autochthonous human case of West Nile fever, 2018 to 2019.

BackgroundDespite the known circulation of West Nile virus (WNV) and Usutu virus (USUV) in Slovakia, no formal entomological surveillance programme has been established there thus far.AimTo conduct contemporaneous surveillance of WNV and USUV in different areas of Slovakia and to assess the geographical spread of these viruses through mosquito vectors. The first autochthonous human WNV infection in the country is also described.MethodsMosquitoes were trapped in four Slovak territorial units in 2018 and 2019. Species were characterised morphologically and mosquito pools screened for WNV and USUV by real-time reverse-transcription PCRs. In pools with any of the two viruses detected, presence of pipiens complex group mosquitoes was verified using molecular approaches.ResultsAltogether, 421 pools containing in total 4,508 mosquitoes were screened. Three pools tested positive for WNV and 16 for USUV. USUV was more prevalent than WNV, with a broader spectrum of vectors and was detected over a longer period (June-October vs August for WNV). The main vectors of both viruses were Culex pipiens sensu lato. Importantly, WNV and USUV were identified in a highly urbanised area of Bratislava city, Slovakias' capital city. Moreover, in early September 2019, a patient, who had been bitten by mosquitoes in south-western Slovakia and who had not travelled abroad, was laboratory-confirmed with WNV infection.ConclusionThe entomological survey results and case report increase current understanding of the WNV and USUV situation in Slovakia. They underline the importance of vector surveillance to assess public health risks posed by these viruses.

Fifteen years since the first record of Trichinella pseudospiralis in Slovakia: What's new?

The presence of Trichinella pseudospiralis has been increasingly reported in Europe in the last decade. The parasite was recorded for the first time in Central Europe in 2003-2004, in eastern Slovakia, in pigs, rats and a cat from a pig breeding farm. In the following years, it was also repeatedly diagnosed in co-infection with T. britovi in sylvatic animals from this area. Molecular analyses revealed a distinctive genetic relationship of the Slovak isolate with those from Finland and Sweden, suggesting the potential role of migratory birds of prey in the transmission of the parasite. Thus, potential host species, including mammals and birds, were investigated for the presence of T. pseudospiralis. During 2006-2018, a total of 360 carcasses of raptorial, carrion-feeding and scavenging birds were collected and examined using artificial digestion of pectoral muscle samples. Muscle larvae were detected in muscle of one golden eagle (Aquila chrysaetos), two common kestrels (Falco tinnunculus) and one peregrine falcon (F. peregrinus). Molecular analyses confirmed the presence of T. pseudospiralis. These findings in diurnal raptorial species represent new host records for Trichinella spp. In 2017, a mandatory examination of pigs (Sus scrofa domesticus), red foxes (Vulpes vulpes) and wild boars (Sus scrofa) revealed one wild boar from the central part of territory to be infected with T. pseudospiralis. Our data confirm that the parasite has already established itself in Slovakia, and thus adequate veterinary measures and public education are needed to prevent its transmission to the food chain and the risk of human infection.

Trichinellosis in Slovakia - epidemiological situation in humans and animals (2009-2018).

INTRODUCTION: Trichinella spp. are zoonotic parasitic nematodes with almost worldwide distribution. The infection can be transmitted through the foodborne route and can cause serious health problems in infected human patients. It is also an economically important issue due to the high financial cost connected with prevention of the disease. OBJECTIVE: The study aimed to discover trends in the epidemiological situation in people and animals in Slovakia in 2009-2018. MATERIAL AND METHODS: Data on human trichinellosis originated from the Public Health Authority of the Slovak Republic, and data on Trichinella infection in animals from the State Veterinary and Food Administration of the Slovak Republic. A seroepidemiological study on 655 voluntary respondents was performed. RESULTS: Altogether, 29 human cases were reported between 2009-2018, with a maximum of 13 cases in 2011. Males were affected more often (19 cases) than females (10 cases); the average age of patients was 45.1 years. Antibodies to Trichinellawere detected in two sera (0.3%): in a serum from one hunter and one veterinarian. In the monitored period, the average prevalence of Trichinella infection was 9.6% in 2,295 red foxes, and 0.04% in 165,643 wild boars. Three (1.7%) of 178 surveyed brown bears were positive. Within the compulsory monitoring of trichinellosis in domestic pigs, none from a total of 1,632,688 pigs were positive. The presence of three species, Trichinellabritovi, T. spiralis and T. pseudospiralis, was documented, with T. britovi representing 93.9% of identified isolates. CONCLUSIONS: The study indicates that the prevalence of Trichinella infection has not changed considerably with time in Slovakia, and the risk of human infection outbreaks is connected mainly with the consumption of wild boar meat.

Ticks (Acari: Ixodidae) Parasitizing Red Foxes (Vulpes vulpes) in Slovakia and New Data About Subgenus Pholeoixodes Occurrence.

BACKGROUND: Distribution and biology of Pholeoixodes ticks is not very well understood. The goal of the study was to collect new data on the Pholeoixodes tick occurrence in Slovakia. METHODS: Tick infestation of red foxes in the regions of Kosice, Presov, Bratislava and Zilina was studied during the period 2017-2018. Ticks were collected from the fur of animals using tweezers and identified using appropriate keys. In total, 146 red foxes (Vulpes vulpes) were investigated. RESULTS: In total, 39 (26.7%) of animals were found to be infected with ticks from five species. Pholeoixodes ticks were found on 13 (3.4%) of the foxes: Ixodes hexagonus (Leach, 1815) on 5 specimens (3.4%), in the Kosice, Presov and Zilina regions; I. crenulatus (Koch, 1844) on 8 specimens (5.5%) in the Presov and Bratislava regions; Ixodes ricinus (Linnaeus, 1758) collected from 25 (17.2%) foxes in every locality; Dermacentor reticulatus (Fabricius, 1794) from 5 foxes (3.4%) in the Kosice, Presov and Zilina regions; Haemaphysalis concinna (Koch, 1844), from 4 foxes (2.8%) from the Kosice region. CONCLUSIONS: Ixodes hexagonus has been previously recorded in Slovakia. However, this is the first finding of I. crenulatus in the country. The morphological features of the I. crenulatus specimens found in Slovakia were identical to those of ticks described in Poland and descriptions given in identification keys.

Seroprevalence of Echinococcus spp. and Toxocara spp. in Invasive Non-native American Mink.

Invasive non-native species can become reservoirs of zoonotic pathogens and cause their spread during colonization, increasing the risk of zoonoses transmission to both wild hosts and humans. American mink (Neovison vison) are considered an important invasive mammal species responsible for carrying endoparasites. The aim of our study was to evaluate the role of feral American mink as a possible transmission vector of Echinococcus spp. and Toxocara spp. in wildlife. We analysed the frequency of American mink exposure to both parasites, the spatial distribution in Poland, and the variability over time on the basis of specific antibody presence using ELISA and Western blot. Alimentary tract analyses revealed that American mink do not serve as definitive hosts for these parasites. Altogether, 1100 American mink were examined. The average seropositivity for American mink was 14.2% for echinococcosis and 21.7% for toxocarosis; dual-seropositivity was detected in only 6.0%. Seroprevalence of both parasites differed between study sites and significantly increased over time in Toxocara spp. Thus, our study revealed that free-living American mink are exposed to parasites and likely to be involved in the maintenance of both Echinococcus spp. and Toxocara spp. in the wild as paratenic hosts.

Co-Circulation of West Nile and Usutu Flaviviruses in Mosquitoes in Slovakia, 2018.

Monitoring West Nile virus (WNV) and Usutu virus (USUV) activity now has the highest priority among mosquito-borne pathogenic viruses circulating in the European Union. This study documents a first time detection and the co-circulation of WNV lineage-2 (with the minimal prevalence of 0.46%) and USUV clade Europe 2 (with the minimal prevalence of 0.25%) in mosquitoes from the same habitat of south-western Slovakia and underlines necessity to perform rigorous surveillance in birds, mosquitoes, horses and humans in that country.

Accumulation of α-2,6-sialyoglycoproteins in the Muscle Sarcoplasm Due to Trichinella Sp. Invasion.

The sialylation of the glycoproteins in skeletal muscle tissue is not well investigated, even though the essential role of the sialic acids for the proper muscular function has been proven by many researchers. The invasion of the parasitic nematode Trichinella spiralis in the muscles with subsequent formation of Nurse cell-parasite complex initiates increased accumulation of sialylated glycoproteins within the affected area of the muscle fiber. The aim of this study is to describe some details of the α-2,6-sialylation in invaded muscle cells. Asynchronous invasion with infectious T. spiralis larvae was experimentally induced in mice. The areas of the occupied sarcoplasm were reactive towards α-2,6-sialic acid specific Sambucus nigra agglutinin during the whole process of transformation to a Nurse cell.The cytoplasm of the developing Nurse cell reacted with Helix pomatia agglutinin, Arachis hypogea agglutinin and Vicia villosa lectin-B4 after neuraminidase pretreatment.Up-regulation of the enzyme ST6GalNAc1 and down-regulation of the enzyme ST6GalNAc3 were detected throughout the course of this study. The results from our study assumed accumulation of sialyl-Tn-Ag, 6`-sialyl lactosamine, SiA-α-2,6-Gal-ß-1,3-GalNAc-α-Ser/Thr and Gal-ß-1,3-GalNAc(SiA-α-2,6-)-α-1-Ser/Thr oligosaccharide structures into the occupied sarcoplasm. Further investigations in this domain will develop the understanding about the amazing adaptive capabilities of skeletal muscle tissue.

Degree of Bacterial Contamination of Mobile Phone and Computer Keyboard Surfaces and Efficacy of Disinfection with Chlorhexidine Digluconate and Triclosan to Its Reduction.

The main aim of our study was to verify the effectiveness of simple disinfection using wet wipes for reduction of microbial contamination of mobile phones and computer keyboards. Bacteriological swabs were taken before and after disinfection with disinfectant wipes with active ingredients chlorhexidine digluconate and triclosan. The incidence and type of microorganisms isolated before and after disinfection was evaluated; the difference was expressed as percentage of contamination reduction. Our results confirmed the high degree of surface contamination with bacteria, some of which are opportunistic pathogens for humans. Before the process of disinfection, on both surfaces, mobile phones, and computer keyboards, the common skin commensal bacteria like coagulase-negative staphylococci were diagnosed most frequently. On the keyboards, species of the genus Bacillus and representatives of the family Enterobacteriaceae were abundant. The potentially pathogenic species were represented by Staphylococcus aureus. Cultivation of swabs performed 5 min after disinfection and subsequent calculation of the reduction of contamination have shown that simple wiping with antibacterial wet wipe led to a significant reduction of microbial contamination of surfaces, with effect ranging from 36.8 to 100%.

Fleas and Ticks of Red Foxes as Vectors of Canine Bacterial and Parasitic Pathogens, in Slovakia, Central Europe.

The present survey aimed to investigate flea and tick fauna parasitizing Slovak red fox populations with special emphasis on canine pathogens they transmit. A total of 407 fleas and 105 ticks were collected from 90 red foxes from two geographically distant regions. Seven flea species (Chaetopsylla globiceps, Pulex irritans, Archaeopsylla erinacei, Chaetopsylla rothschildi, Chaetopsylla trichosa, Ctenocephalides canis, and Ctenopthalmus assimilis) and three species of hard ticks (Ixodes ricinus, Ixodes hexagonus, Haemaphysalis concinna) were recorded on sampled animals. Consequently, the DNA of five different pathogen taxa was confirmed in collected arthropod vectors: Bartonella spp. (in P. irritans, Ch. globiceps, and Ct. assimilis), Rickettsia spp. (in A. erinacei, I. ricinus, I. hexagonus, and H. concinna), Anaplasma phagocytophilum (in I. ricinus), Theileria sp. (in Ch. globiceps and H. concinna), and Hepatozoon canis (in I. ricinus and I. hexagonus). Mycoplasma spp., Dipylidium caninum, and Acanthocheilonema reconditum were not found in fleas or ticks in this study.

Update of the helminth fauna in Eurasian lynx (Lynx lynx) in Poland.

The Eurasian lynx (Lynx lynx) is a strictly protected species of large carnivore in Poland. It inhabits forest complexes in north-eastern Poland and the Carpathian region in southern Poland. The status of the lynx within Poland requires special attention because its range decreased between 1980 and 2001 and has not yet recovered. One of the factors negatively affecting lynx populations is diseases, particularly parasites. The helminth fauna of the Eurasian lynx is not fully known in Poland. Previous coprological studies revealed that Polish lynx have been infected with seven species of nematodes, three species of cestodes, and one species of trematode. In this study, we present new data based on examination of opportunistically collected lynx carcasses. The aim of the study was to complement data on the helminth fauna of Eurasian lynx inhabiting Poland based on morphological and molecular analysis of parasites. Four species of cestodes-Taenia lynciscapreoli, Mesocestoides lineatus, Spirometra sp., and Taenia krabbei-were found for the first time in Eurasian lynx from Poland and three previously reported species of nematodes-Ancylostoma tubaeforme, Toxascaris leonina, and Toxocara cati-were confirmed. Larvae of Trichinella britovi were also detected in Eurasian lynx in Poland for the first time.

Urbanization impact on mosquito community and the transmission potential of filarial infection in central Europe.

BACKGROUND: Despite long-term research on dirofilariosis in Slovakia, little attention has thus far been paid to Dirofilaria vectors. The particular aim of the present study was molecular screening for filarioid parasites in two different habitats of Bratislava, the capital city of Slovakia. In addition, the effect of urbanisation on mosquito species abundance and composition, associated with the risk of mosquito-borne infections, was studied and discussed. METHODS: Mosquitoes were identified by morphological features, and molecular methods were also used for determination of selected individuals belonging to cryptic species from the Anopheles maculipennis and Culex pipiens complexes. The presence of filarioid DNA (Dirofilaria repens, Dirofilaria immitis and Setaria spp.) was detected using standard PCR approaches and sequencing. RESULTS: A total of 6957 female mosquitoes were collected for the study. Overall, the most abundant mosquito species was Aedes vexans, closely followed by unidentified members of the Cx. pipiens complex and the less numerous but still plentiful Ochlerotatus sticticus species. Further investigation of mosquito material revealed 4.26% relative prevalence of Dirofilaria spp., whereby both species, D. repens and D. immitis, were identified. The majority of positive mosquito pools had their origin in a floodplain area on the outskirts of the city, with a relative prevalence of 5.32%; only two mosquito pools (1.26%) were shown to be positive in the residential zone of Bratislava. Setaria spp. DNA was not detected in mosquitoes within this study. CONCLUSIONS: The study presented herein represents initial research focused on molecular mosquito screening for filarioid parasites in urban and urban-fringe habitats of Bratislava, Slovakia. Molecular analyses within the Cx. pipiens complex identified two biotypes: Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus. To our knowledge, Dirofilaria spp. were detected for the first time in Slovakia in mosquitoes other than Ae. vexans, i.e. D. repens in Anopheles messeae and unidentified members of An. maculipennis and Cx. pipiens complexes, and D. immitis in Coquillettidia richiardii and Cx. pipiens biotype pipiens. Both dirofilarial species were found in Och. sticticus. The suitable conditions for the vectors' biology would represent the main risk factor for dirofilariosis transmission.

GIS-based environmental analysis of fox and canine lungworm distribution: an epidemiological study of Angiostrongylus vasorum and Crenosoma vulpis in red foxes from Slovakia.

Over a period of intervening years, the distribution of two canine cardiopulmonary metastrongylid nematodes, Angiostrongylus vasorum and Crenosoma vulpis, has been recognised in Central Europe. Here, we report the first epidemiological research conducted in red foxes from Slovakia and the potential influence of selected environmental variables on the parasites' occurrence, quantified by logistic regression. The environmental models revealed that distribution of C. vulpis is not significantly influenced by any environmental variables, and the parasite is present in the whole area under study. Models for A. vasorum revealed some weak influence of environmental variables, as it tends to occur in drier areas with lower proportion of forest. Moreover, A. vasorum shows a typical spatial clustering and occurs in endemic foci identified mainly in the eastern part of Slovakia. A cluster of A. vasorum infection foci was also found in the north-eastern region, where the average winter air temperature regularly falls below - 10 °C.