The first report of Setaria tundra (Issaitshikoff & Rajewskaya, 1928) in Slovakia by using of molecular methods.

Setaria tundra is a filarioid parasite occurring in the northern hemisphere. Adult forms of helminths are located free in the peritoneal cavity of its definitive host - cervids, while microfilariae are presented in the host's bloodstream. Intermediate hosts are represented by several mosquito species, mainly of the genus Aedes.Nematode S. tundra is well adapted to roe deer (Capreolus capreolus) and therefore is the infection usually asymptomatic. In this study we present the first report of S. tundra in Slovakia. During a period 2022 a total of 6 roe deer coming from eastern Slovakia (Trebisov district) were examined. Nematodes were found during the evisceration process in the abdominal cavity of 3 specimens Intensity of infection was in range from 5 to 38 helminths per host. Mean intensity of infection reached 18.3 parasites per host. The helminths were identified as S. tundra by morphological examination and molecular typing of the COI gene. This study is the first report of S. tundra in Slovakia.

Morphological and molecular identification of adult Stichorchis subtriquetrus in beaver in Slovakia.

The European beaver (Castor fiber) was a fully eradicated species in Slovakia. Thanks to year-round protection and several different reintroduction programs the population is now increasing. However, there are limited reports about their health status.A 2-year-old female European beaver, was found dead by road near town Hanusovce nad Toplou in eastern Slovakia in 2021. Necropsy was carried out at the Department of Breeding and Diseases of the Game, Fish and Bees, Ecology and Cynology, University of Veterinary Medicine and Pharmacy in Kosice. Necropsy revealed a total of 13 trematodes, collected from the caecum and colon. Based on morphological and molecular analysis digenean trematode Stichorchis subtriquetrus (Rudolphi, 1814) was identified. This is the first record of adult helminth Stichorchis subtriquetrus in beaver in Slovakia.

The survey of soil-transmitted helminth species abundance in Slovakia with an emphasis on parameters important for their distribution.

Soil-transmitted helminths (STH) can be easily dispensable in socially disadvantaged groups. The Roma people represent the group most at risk in Slovakia. This study aimed to investigate the occurrence of STH infections in minorities living with animals under low hygienic conditions and on contaminated soil. Subsequently, we identified the risk assessment factors of the STH transmissions based on parasitological results. In our study, STHs were predominantly found among the Roma communities living in unsanitary conditions. The prevalence of Ascaris lumbricoides among the majority was 0.79%, and Trichuris trichiura was 0.05 %. On the contrary, a community-based cross-sectional survey across eastern Slovakia also found a prevalence of 22.28 and 3.47% for A. lumbricoides and T. trichiura among the Roma population. Inhabitants that belong to the Roma minority had a 37.12 infection OR times higher than non-Roma inhabitants. The Roma people living in the countryside have a 2.23-fold higher chance of getting infected with STH than Roma living in the city. Therefore, soil and domestic animals were also examined for the presence of the STH eggs to show the environmental burden. In general, the presence of STH eggs was confirmed in 26.26% of the soil examined samples. The detailed description is as follows: eggs of Toxocara spp., Trichuris spp., eggs from the family Ancylostomatidae, and Toxascaris leonina. Ascaris eggs were detected only in the soil from localities with low hygienic standards. The probability of contracting the STH eggs in segregated settlements was 15.94 times higher compared to urban or rural areas. In addition to humans, dogs can also be a source of STH eggs in the soil. The STH eggs were confirmed in 43.55% of dog droppings. The most interesting finding was that the eggs of the genus Ascaris were up to 7.93% of dog droppings from localities with a low hygienic standard were positive. This study revealed that climatic factors and the WASH conditions influenced the distribution of STHs to variable degrees. In addition, ethnicity and sanitation were crucial factors in the distribution of STH infection in eastern Slovakia.

Effect of Wastewater Treatment on Bacterial Community, Antibiotic-Resistant Bacteria and Endoparasites.

Wastewater and wastewater treatment plants serve as urban reservoirs of pathogenic microorganisms. Wastewaters frequently contain bacteria, antibiotic-resistant bacteria, and developmental stages of parasites with significant zoonotic potential. Five wastewater treatment plants in the central part of Slovakia were investigated to determine the effect of treatment on bacterial community, antibiotic-resistant bacteria, and the occurrence of helminth eggs. Although all monitored chemical factors (chemical oxygen demand, biochemical oxygen demand, N-NH4, total nitrogen, and total phosphorus) in the effluent were in line with the legislative standards for discharge into public waterways, the results of minimal inhibitory concentrations show that reclaimed water harbors E. coli resistant to several commonly used antibiotics (ampicillin, piperacillin, and tazobactam, combine ampicillin and sulbactam, cefotaxime, tetracycline). The presence of endoparasite developmental stages in wastewater and sludge (Ascaris spp., Hymenolepis nana, eggs from the Ancylostomatidae family, Giardia duodenalis) indicates potential health risks for humans and workers at these sites. Treatment such as composting before applying sludge to land is necessary to reduce human pathogens.

Neglected Diseases-Parasitic Infections among Slovakian Children from Different Populations and Genotypes of Giardia duodenalis.

Children are most prone to parasitic infections. The objectives of the study were to examine the occurrence of parasitic infections in children from different populations and to perform molecular characterization of human Giardia duodenalis isolates. We examined 631 stool samples from Roma and non-Roma children for the presence of parasitic developmental stages. Samples were collected from three eastern Slovakia districts. The ages of the children ranged from 1 months to 17 years. Subsequently, the molecular characterization of human G. duodenalis isolates by PCR detected triosephosphate isomerase (tpi) and beta-giardin (bg) genes was performed. The overall prevalence of parasitic infection was 19.8%. Ascaris lumbricoides eggs were the most frequent, with an occurrence of about 13.8%. G. duodenalis cysts were present in 6.3% of samples. G. duodenalis isolates obtained from 13 children were subjected to DNA sequencing with tpi and bg genes. Five isolates were categorized as bearing subassemblage BIII, the three isolates as subassemblage BIV, one person was infected with a mixture of subassemblages BIII and BIV, four children had subassemblage AII, and one isolate revealed a structure corresponding with subassemblage AI. Our work is proof that poverty and poor hygiene contribute the most to public health problems associated with neglected parasitic diseases.

Genotyping of Ascaris spp. infecting humans and pigs in Italy, Slovakia and Colombia.

BACKGROUND: The systematics and taxonomy of Ascaris lumbricoides and Ascaris suum, two of the world's most widespread nematodes, still represent a highly debated scientific issue. Two different transmission scenarios have been described according to endemicity: separated host-specific transmission cycles in endemic regions, and a single pool of infection shared by humans and pigs in non-endemic regions. The swine roundworm A. suum is now recognized as an important cause of human ascariasis also in endemic areas such as China, where cross-infections and hybridization have also been reported, as well as in non-endemic regions like Italy. This study aimed to investigate the molecular epidemiology of human and pig ascariasis in three countries representing different epidemiological scenarios: Italy as a non-endemic country, Colombia as an endemic country, and Slovakia as a non-endemic country, but with a poor socio-economic context linked to some focal populations of Roma settlements. MATERIALS AND METHODS: A total of 237 nematodes were analysed: 46 from Colombia (13 from humans, 33 from pigs), 114 from Slovakia (20 from humans, 94 from pigs) and 77 from Italy (17 from humans and 60 from pigs). Genotyping by PCR-RFLP of nuclear (ITS) and sequencing of mitochondrial (cox1) target regions were performed. ITS genotypes were used to estimate the Hardy-Weinberg (HW) equilibrium according to hosts and country of origin. The partial cox1 sequences were used to analyse genetic polymorphisms according to hosts and country of origin, as well as to infer the network of haplotypes, their evolutionary relationships and geographical distribution. RESULTS: 110 quality cox1 sequences were obtained. Haplotype network revealed three main groups corresponding to clade A, B and C. Clade C included most of the human cases from Italy, while those from Slovakia and Colombia were grouped in clade B. Ascaris from Italian and Colombian pigs showed HW equilibrium at the ITS marker, while disequilibrium was found in A. lumbricoides from Slovak pigs, which suggest a high unexpected amount of roundworms of human origin circulating also in pigs. CONCLUSIONS: This study updates and extends the current understanding of Ascaris species and genotypes circulating in different epidemiological scenarios, with particular attention to the inclusion of human-derived Ascaris in the phylogenetic cluster C. Despite the evidence of HW equilibrium in the ITS in pig-derived Italian samples, the amount of genetic variation seems to support the existence of two closely related species.

The occurence of endoparasites in Slovakian household dogs and cats.

Pets play a pivotal role as definitive or reservoir hosts for many zoonotic parasites. Dogs and cats without any clinical signs may be a carrier for the infection. In a one-year study, collected fecal samples of 257 dogs and 50 cats were examined coproscopically for endoparasite infections. Out of 307 investigated fecal samples, 107 (34.9%) were positive for the presence of the propagative stages of endoparasites In 257 dogs fecal samples, following 12 different species of endoparasites were detected: Giardia duodenalis, Cystoisospora spp., Sarcocystis spp., Hammondia/Neospora-like eggs, Angiostrongylus vasorum larvae, Capillaria aerophila, Crenosoma vulpis, Toxocara spp., Toxascaris leonina, Trichuris vulpis, Strongyloides stercoralis, and eggs from the family Ancylostomatidae. Only 4 different parasitic species were found in 50 domestic cat fecal samples - G. duodenalis cysts Cystoisospora spp., T. cati, and larvae of Aelurostrongylus abstrusus. It was confirmed that significant differences were found concerning age for G. duodenalis, T. canis, S. stercoralis, and family Ancylostomatidae. Close and frequent contact between pets and people increases the risks for the transmission of zoonotic diseases.

First report on parasites of European beavers in the Slovak Republic.

European beaver (Castor fiber L. 1758) is the biggest rodent living in Europe. It is a semi-aquatic animal known for building dams and burrows. European beaver is a potential host for a wide range of parasites and other infectious diseases. In Slovakia, there is an increasing number of beavers but the data about their parasitic fauna are missing. Our work is the first documentation about the beaver's parasitofauna in Slovakia. In a 1-year study, we collected and examined 19 beaver fecal samples from the vicinity of beaver burrows inhabiting three particular localities at the Danube, Topla, and Laborec rivers in Slovakia. In these fecal samples, 4 different species of intestinal endoparasites were detected as follows: oocysts of Cryptosporidium, cysts of Giardia, eggs of Stichorchis subtriquetrus, and eggs and larvae of Travassosius rufus. Parasites were confirmed only in samples collected at river Topla. Based on our results, we can conclude that European beaver can be an important source of parasitic contamination of surface waters especially in the localities shared by people.

Intestinal parasites in segregated minority communities of Slovakia: results from a cross-sectional survey in children.

In Slovakia, the Roma population forms the second-largest ethnic minority. A large part of the Roma is semi-nomadic or lives in segregated settlements with poor living standards and limited access to health facilities. More than 40 years ago, a cross-sectional survey revealed a high prevalence of parasitic infections. There is a paucity of recent data, and hence, we designed a study to investigate the current status of intestinal parasitic infections in this population. Overall, 259 children aged 7 months to 18 years from 32 different segregated settlements provided faecal samples for microscopic examination using a sodium acetate-acetic acid-formalin concentration and the Paraprep L technique. Almost 40% of the samples yielded a positive result, with Ascaris lumbricoides (27.4%) and Giardia intestinalis (9.3%) being the most frequent helminth and intestinal protozoa species, respectively. Many children younger than 2 years were found to be infected, which suggests that community transmission is important. In view of our findings, there is a pressing need for targeted action to improve the health status of this neglected population.

First report on Giardia duodenalis assemblage F in Slovakian children living in poor environmental conditions.

BACKGROUND: Giardiasis is one of the most common gastrointestinal infections of humans and animals attributable to complex of eight morphologically identical genetic assemblages, further divided into sub-assemblages. Disease is common for a wide range of hosts and genetic characterization is needed for better understanding of multifaceted epidemiology for this protozoan parasite. The aim of this study was to identify genetic heterogeneity in assemblages and sub-assemblages of Giardiaduodenalis circulating among the children population living in deprived socioeconomic conditions. METHODS: A total of 333 stool samples from children in eastern Slovakia were collected during the period of 2015-2016 and analysed by molecular methods. Molecular characterization of G. duodenalis was performed by sequence analysis of triose phosphate isomerase gene (tpi) and glutamate dehydrogenase gene (gdh). RESULTS: G. duodenalis DNA was detected in 21 samples (6.3%), out of which 14 isolates (66.7%) belonged to assemblage B, 4 isolates (19.0%) to sub-assemblage AII and 3 isolates (14.3%) corresponded to assemblage F. As regards the determination of sub-assemblages of assemblage B, 4 isolates were characterized as sub-assemblage BIII and 6 isolates as sub-assemblage BIV. CONCLUSION: This study is the first finding of cat specific assemblage F in man not only in Slovakia, but also in Europe. The absence of molecular data about G. duodenalis in companion animals in Slovakia establishes a strong need for further investigation for potential sources of giardiasis and understanding the epidemiology will help to improve the preventive strategies in eradication of infection in this population.

Devitalisation of pathogens in stored pig slurry and potential risk related to its application to agricultural soil.

The study investigated the risks arising from application of pig slurry to soil in relation to viability of Salmonella typhimurium, Escherichia coli, total coliforms, faecal enterococci and eggs of Ascaris suum at different temperatures. Potential effect of changes in physico-chemical parameters, particularly dry matter (DM), pH and ammonia, were also investigated. Examination showed that S. typhimurium was devitalised after storage in the slurry for 115 days at 4 °C and after 90 days at 20 and 42 °C. Devitalization of E. coli and faecal entrerococci required more than 115 at temperature of 4 °C and faecal enterococci were recovered from slurry after 115 days of storage even at temperature of 20 °C. Total coliforms survived for 115 days at all investigated temperatures. Complete devitalization of A. suum eggs was not achieved even after 115 days at 42 °C. Our investigations indicated potential microbial and parasitic risk related to application of pig slurry to soil even after 115 days of storage.

Occurrence of the most common helminth infections among children in the Eastern Slovak Republic.

OBJECTIVES: Ascariasis, trichuriasis and hymenolepiasis occur primarily within poor communities with low hygiene standards. This study examined the occurrence of intestinal helminth infections among children living in two counties (Kosice and Presov) in the Eastern Slovak Republic. STUDY DESIGN: Four hundred and twenty-six children were divided into groups according to ethnicity (non-Roma and Roma), age, sex, urban/rural residency and county of residence. METHODS: Stool samples collected from participants were processed by formalin-ethyl acetate sedimentation and examined microscopically. RESULTS: The overall prevalence of infection was 16.90% and the most prevalent species was Ascaris lumbricoides (14.32%). This was followed by Trichuris trichiura (3.76%), Hymenolepis nana (0.94%) and Hymenolepis diminuta (0.23%). The odds ratio for infection was 52 times higher among Roma children compared with non-Roma children. Among Roma children, the lowest prevalence of infection was reported in adolescents aged ≥16 years. No significant differences in the prevalence of helminths were found between different sexes, or between hospitalized and non-hospitalized participants. Roma children living in urban areas had a 3.36 higher probability of infection than those living in rural areas. Among Roma children, helminth ova were found in 31.76% of the specimens from Kosice County and 19.69% of the specimens from Presov County. Among non-Roma children, there was only one positive finding in Presov County, and no cases in Kosice County. CONCLUSIONS: Important risk factors associated with helminth infections are ethnicity, county of residence and urban/rural residency. Ascariasis, trichuriasis and hymenolepiasis still occur in children with low hygiene standards, and this needs to be addressed by local authorities.

Characterization of composting mixtures and compost of rabbit by-products to obtain a quality product and plant proposal for industrial production.

In this study we have observed the effects of using rabbit manure and slaughtering by-products in a composting process. Three piles of this material, 4700 kg each, with different amount and C/N ratio, have been investigated and experimental tests were carried out in an industrial horizontal axe reactor using a prototype of turning machine. The composting time lasted 85 days; 2 experimental cycles were conducted: one in Winter and one in Summer. In the Winter test, mesophilic reaction started only in the control mixture (animal manure + slaughtering by-products without straw). It is noteworthy that, the 3 investigated mixtures produced soil amendment by compost with good agronomical potential but with parameters close to the extreme limits of the law. In the Summer test, there was thermophilic fermentation in all mixtures and a better quality compost was obtained, meeting all the agronomic and legislative constraints. For each pile, we examined the progression of fermentation process and thus the plant limitations that did not allow a correct composting process. The results obtained in this study are useful for the development of appropriate mixtures, machines, and plants assuring continuance and reliability in the composting of the biomass coming from rabbit industry.

Nematodes as bioindicators of soil degradation due to heavy metals.

The effect of distance from a heavy metal pollution source on the soil nematode community was investigated on four sampling sites along an 4 km transect originating at the Kovohuty a.s. Krompachy (pollution source). The soil nematode communities were exposed to heavy metal influence directly and through soil properties changes. We quantified the relative effects of total and mobile fraction of metals (As, Cd, Cr, Cu, Pb, and Zn) on soil ecosystem using the nematode community structure (trophic and c-p groups,) and ecological indices (Richness of genera, H', MI2-5, etc.). Pollution effects on the community structure of soil free living nematodes was found to be the highest near the pollution source, with relatively low population density and domination of insensitive taxa. A decrease in heavy metals contents along the transect was linked with an increase in complexity of nematode community. The majority of used indices (MI2-5, SI, H') negatively correlated (P < 0.05 or P < 0.01) with heavy metals content and were sensitive to soil ecosystem disturbance. Contamination by heavy metals has negatively affected the soil environment, which resulted in nematode community structure and ecological indices changes. Results showed that the free-living nematodes are useful tools for bioindication of contamination and could be used as an alternative to the common approaches based on chemical methods.

Sanitation composting process in different seasons. Ascaris suum as model.

The influence of aerobic composting of organic wastes (straw, sawdust, wood brash and sludge from WWTP) on the viability of the model parasite eggs were studied in production conditions of the Industrial Composting Plant TOPVAR, Inc. in Topolcany, Slovak Republic. During summer and winter seasons (2005-2006), the survival of non-embryonated Ascaris suum eggs was evaluated with respect to aerobic composting processes and air temperature, which varied from -6 degrees C to +12 degrees C in winter and from 11 degrees C to 32 degrees C in summer. Our results revealed the induction of maximum temperature of the substrate (+65 degrees C) on day 6 of the composting irrespective of the low air temperature in winter. During the summer season, higher substrate temperature was observed (+71 degrees C). Due to a high temperature and chemical changes (pH, dry matter, inorganic and organic matter, NH(4)(+), total phosphorus, total nitrogen and the C:N ratio), which occur in substrate during composting process, the total devitalization of the A. suum eggs was observed between day 4-5 of the experiment in the winter season and on day 6 in the summer season. Thus we may conclude that the final product is suitable for agricultural purposes without a risk of transmitting the pathogenic germs into the environment.