Molecular screening for enteric parasites and subtyping of Blastocystis sp. in haemodialysis patients in Slovakia.

INTRODUCTION AND OBJECTIVE: Intestinal parasitoses are important causes of morbidity and mortality, especially in immunocompromised individuals. In patients with chronic renal insufficiency (CRI), the accumulation of non-excreted metabolites leads to uraemia, which induces a state of immunodeficiency, increasing the incidence of infections. The aim of the study was molecular screening for enteric protozoa in patients with chronic renal insufficiency. MATERIAL AND METHODS: A total of 53 samples were collected in January 2023 from patients undergoing dialysis at Logman Ltd. Nephrodialysis Centre in Kosice, Slovakia. Samples were examined by polymerase chain reaction (PCR) for the presence of Cryptosporidium parvum / Cryptosporidium hominis, Giardia intestinalis, Microsporidia spp., and Blastocystis sp. RESULTS: From the 53 samples, the only pathogen identified by PCR was Blastocystis sp., in 13 patients (24.5 %). Sequence analyses confirmed that the most prevalent subtype (ST) among patients was ST 3 (n=9, 69.2%), followed by ST 1 (n=3, 23.1%) and ST 2 (n=1, 7.7%). CONCLUSIONS: Molecular methods for the detection of microscopic enteric parasites are not used as a first-line diagnostic method in Slovakia. In immunocompromised patients, diarrhoea can be caused not only by a chronic disease or therapy but can also be a result of an ongoing underdiagnosed infection. Early diagnosis leads to targeted therapy and subsequent partial improvement of the quality of life. This study also shows the first insights into Blastocystis sp. subtype distribution in humans in Slovakia.

Is Slovakia Almost a Hepatitis D Free Country?

BACKGROUND: It is assumed that the prevalence of hepatitis D in HBsAg-positive individuals reaches 4.5-13% in the world and on average about 3% in Europe. Data from several European countries, including Slovakia, are missing or are from an older period. METHODS: We analyzed all available data on hepatitis D from Slovakia, including reports from the Slovak Public Health Authority and the results of one prospective study, and three smaller surveys. The determination of anti-HDV IgG and IgM antibodies and/or HDV RNA was used to detect hepatitis D. RESULTS: In the years 2005-2022, no confirmed case of acute or chronic HDV infection was reported in Slovakia. The presented survey includes a total of 343 patients, of which 126 were asymptomatic HBsAg carriers, 33 acute hepatitis B, and 184 chronic hepatitis B cases. In a recent prospective study of 206 HBsAg-positive patients who were completely serologically and virologically examined for hepatitis B and D, only 1 anti-HDV IgG-positive and no anti-HDV IgM or HDV RNA-positive cases were detected. In other smaller surveys, two anti-HDV IgG-positive patients were found without the possibility of HDV RNA confirmation. In total, only 3 of 329 HBsAg-positive patients (0.91%) tested positive for anti-HDV IgG antibodies, and none of 220 tested positive for HDV RNA. CONCLUSION: The available data show that Slovakia is one of the countries with a very low prevalence of HDV infection, reaching less than 1% in HBsAg-positive patients. Routine testing for hepatitis D is lacking in Slovakia, and therefore it is necessary to implement testing of all HBsAg-positive individuals according to international recommendations.

Vaccination, Risk Factors and Outcomes of COVID-19 Infection in Patients with Psoriasis-A Single Centre Real-Life Experience from Eastern Slovakia.

Coronavirus disease (COVID-19) represents a threat for people with immune-mediated diseases. It seems that patients with psoriasis appear to have a similar SARS-CoV-2 infection rate as the general population. Our study aimed to identify factors associated with contracting COVID-19 and determining the severity of COVID-19 among psoriatic patients in a real practice setting. We conducted a cross-sectional study with 379 respondents. About one-quarter (n = 78; 25.8%) of the respondents who provided information on their COVID-19 (n = 302) status had contracted COVID-19. Most variables tested for their effect on getting COVID-19 proved to be statistically insignificant, except education, age and gender. Our study proved the protective effect of vaccination, especially the third dose, against the COVID-19 outcome. From all the potential variables, we found that non-Roma ethnicity increased the chance of being vaccinated at least once by 2.6-fold. Patients with a longer psoriasis duration had a higher chance of being vaccinated. We consider biological treatment of psoriasis safe during COVID-19. Vaccination of patients was a statistically significant protector against COVID-19. It is important to point out that only three doses of vaccine decreased with statistical significance the chance of getting the illness. Our findings should be confirmed on larger samples in further studies.

Occurrence of cryptosporidium parvum IIaA17G1R1 in hospitalized hemato-oncological patients in Slovakia.

This study aimed to determine the presence of cryptosporidiosis in immunosuppressed patients hospitalized at the Clinic of Haematology and Oncohaematology in a form of routine screening. Samples were collected from November 2019 to February 2020, when the first wave of the Coronavirus pandemic occurred in Slovakia. A total of 36 samples were collected from patients hospitalized at the Clinic of Haematology and Oncohaematology, both from the open ward and the intensive care unit. For the diagnosis of cryptosporidiosis, a nested PCR targeting the gp60 gene and the SSU rRNA locus was used. From the 36 samples, Cryptosporidium parvum subtype IIaA17G1R1 was diagnosed in 9 patients (7 from the open ward and 2 from the intensive care unit), all hospitalized at the clinic at the same time, in February 2020. The occurrence of the same species and subtype, Cryptosporidium parvum IIaA17G1R1, in 9 patients hospitalized at the same time, both at the open ward and the intensive care unit may suggest a possible transmission occurred at the clinic.

Comparison of Two Diagnostic Methods for the Detection of Hepatitis B Virus Genotypes in the Slovak Republic.

The hepatitis B virus (HBV), belonging to the Hepadnaviridae family, is responsible for a global health concern still in the 21st century. The virus is divided into 10 genotypes, which differ in geographical distribution and in their effect on disease progression and transmission, susceptibility to mutations, and response to treatment. There are many methods for diagnostics of HBV and differentiating its genotypes. Various commercial kits based on real-time polymerase chain reaction (RT PCR) and hybridization available, as well as whole genome sequencing or the sequencing of only individual parts of the genomes. We compared a commercial kit AmpliSens HBV-genotype-FRT, based on RT PCR, with an adapted method of amplification of the surface genomic region combined with Sanger sequencing. In the examined samples we identified the A, B, C, D, and E genotypes. By PCR with Sanger sequencing, the genotypes were determined in all 103 samples, while by using the commercial kit we successfully genotyped only 95 samples, including combined genotypes, which we could not detect by sequencing.

The first report of animal genotypes of Cryptosporidium parvum in immunosuppressed and immunocompetent humans in Slovakia.

The aim of our study was to determine species and genotypes of Cryptosporidium in patients suffering from immunosuppressive illnesses, but also in immunocompetent patients suffering from diarrhoea. A total of 80 samples of faeces were collected from both immunosuppressed and immunocompetent patients. The immunosuppressed patients (65 samples) - 35 adult patients (group A) and 30 children (group B) were hospitalized at the Clinic of Oncohemathology. Samples from immunocompetent humans (15 samples, group C) were taken from patients with clinical signs of acute diarrhoea. With the use of molecular methods targeting the 60 kDa glycoprotein (GP60) gene region, we have identified multiple genotypes of Cryptosporidium. parvum and Cryptosporidium. hominis in immunocompromised, but also in immunocompetent individuals (C. hominis IbA10G2, IeA12G3T3; C. parvum IIaA10G1R1, IIaA11G2R1, IIaA12G2R1, IIaA13G1R1, IIaA14G1R1, IIaA14G2R1, IIaA17G1R1 and IIaA18G1R1). This is the first report of the occurrence of genotypes IIaA10G1R1, IIa12G2R1 and IIaA18G1R1 in human hosts.

Concentrations of selected toxic elements in ewe living near an environmentally loaded area of eastern part of Slovakia.

INTRODUCTION AND OBJECTIVE: Research focused on the monitoring of selected heavy metals in ewes' blood. Concentrations of selected toxic elements, lead, cadmium and zinc, in ewes living near an environmentally-loaded area, concerned 15 ewes (aged 3-4 years) in good physical condition, during the spring of 2014 and 2015 in the eastern part of Slovakia. The aim of the research was to determine the concentration of selected heavy metals and state the correlations of selected heavy metals in ewes' blood. MATERIAL AND METHODS: Within the period of 2 years, 15 ewe were evaluated. Ewes' blood samples were collected twice during the spring season from a farm located in area Spis, eastern Slovakia, and then analysed for heavy metal contents. In the area under investigation, contamination with heavy metals was assumed as a result of intensive agricultural development and former mining activities. The level of selected heavy metals in the experimental group of animal blood was determined using an optical spectrophotometry. RESULTS: Statistical analyses were carried out using the Statistica programme. The significant differences between means were calculated by the statistical method of the non-parametric Mann-Whitney´s U test. The statistical test experimental group of ewes in 2014 and 2015 confirmed the presence of selected heavy metals in ewes. The measured values of Cd (P=0.0003), Pb (P=0.0200) and Zn (P=0.0018) showed significant differences when comparing the years 2014 and 2015. CONCLUSIONS: The obtained and analysed blood samples confirmed the presence of selected heavy metals in ewes from area of Spis in eastern Slovakia, which belongs the sub-region or is among the localities environmentally burdened. The conclusions are centred on the population's interest and concern for the environment, as well as on the preoccupation with factors that affect the satisfaction of basic needs, the local agricultural development and former mining activities.

Rodents as a reservoir of infection caused by multiple zoonotic species/genotypes of C. parvum, C. hominis, C. suis, C. scrofarum, and the first evidence of C. muskrat genotypes I and II of rodents in Europe.

Cryptosporidium spp. is an important causative agent of intestinal parasitoses-induced diarrhoea in humans and animals worldwide. Rodents (small mammals), the main reservoir of infections, are globally expanded and overpopulated, which increases the risk of transfer of human and zoonotic pathogens from the genus Cryptosporidium. In this study, Cryptosporidium was detected in wild immunocompetent asymptomatic small mammals. Altogether 262 fecal samples were collected from five areas in Eastern Slovakia from four different rodent species (Myodes glareolus, Apodemus agrarius, Apodemus flavicollis, Rattus norvegicus), eight samples originated from two insectivore species (Sorex araneus, Crocidura suaveolens), and two sample from a carnivore Mustela nivalis. The samples were examined using a method modified in our laboratory, based on the use of specific primers on a small subunit rRNA (18S rRNA) gene for species identification, and amplification of GP60 gene coding 60-kDa glycoprotein for genotype determination. The following species were identified: Cryptosporidium parvum (n=15), genotypes IIaA18G3R1 (n=11; KU311673), IIaA10G1R1 (n=1; KU311670), IIcA5G3a (n=1; KU311669), IIiA10 (n=2; KU311672); Cryptosporidium suis (n=4; KU311671); Cryptosporidium scrofarum (n=28); Cryptosporidium environment sp. (n=12; KU311677); Cryptosporidium muskrat genotype I (n=3; KU311675); Cryptosporidium muskrat genotype II (n=3; KU311676). From one of the rodent, the species Cryptosporidium hominis genotype IbA10G2 (KU311668) was identified for the first time. The results of this study indicate low host specificity of the detected Cryptosporidium species and imply the importance of free-living small mammals in urban and suburban habitats as a potential source of human cryptosporidiosis.