Surveillance Program of Clinical Samples for Polio and Non-Polio Enteroviruses in the Slovak Republic during the 1958-2020 Period.

Enteroviruses (EVs) are associated with a wide spectrum of diseases involving various organs. Our aim was to give a historical overview of the genesis of clinical sample processing for EVs in the Slovak Republic (SR) during the 1958-2020 period, within the framework of the World Health Organization (WHO) polio program. Further, analyses were made of the data obtained from the archives of processed clinical sample surveillance using statistical methods. We used generalized additive models (GAM) with binomial distribution and logit link functions and an autoregressive moving average (ARMA) to analyze the data obtained during this 63-year period. Our results show trends in the composition of EV strains circulating in the population. Furthermore, statistically significant increasing trends of the non-polio enteroviruses (NPEVs) were observed over the studied time, represented by echoviruses (E) and coxsackieviruses A and B (CVA and CVB), with a cyclical pattern of occurrence. The most prevalent serotype over this period was CVB5, which became significantly more prevalent after 2000. While PVs, CVB1, and CVB3 were present in the second half of the studied period, CVA10, CVA16, E3, E25, and E30 appeared more frequently.

The stringency of the containment measures in response to COVID-19 inversely correlates with the overall disease occurrence over the epidemic wave.

Non-pharmaceutical interventions (NPIs) were the only viable choice to mitigate or suppress transmission of COVID-19 in the absence of efficient and safe vaccines. In this study, we examined the association between the stringency of containment measures and cumulative incidence of the COVID-19 cases in the first wave of the pandemic across 28 European countries. Our results support the effectiveness of containment measures in the mitigation or suppression of COVID-19 epidemics. Early adoption of stringent containment measures prior to detection of the first confirmed case, together with ramping up containment stringency during the early days of epidemics, was associated with a lower disease occurrence. The delayed adoption of stringent containment measures did not fully compensate for the lack of early response. Containment measures continue to play a significant role in the control of COVID-19 in the post-vaccination period, when limited vaccination coverage, the emergence of vaccine resistance, and/or increased mobility enabled further disease transmission (Tab. 4, Fig. 22, Ref. 50). Keywords: non-pharmaceutical interventions, containment, COVID-19, mobility, social distancing, Containment and Health Index, epidemiology, public health measures, SARS-CoV-2.

History of the Wastewater Assessment of Polio and Non-Polio Enteroviruses in the Slovak Republic in 1963-2019.

We describe the genesis of poliovirus (PV) and non-polio enterovirus (NPEV) surveillance program of sewage wastewaters from its inception to the present in the Slovak Republic (SR). Sampling procedures and evolution of the methodology used in the SR for the detection of PVs and NPEVs are presented chronologically. For statistical data processing, we divided our dataset into two periods, the first period from 1963 to 1998 (35 years), and the second period from 1999 to 2019 (21 years). Generalized additive models were used to assess temporal trends in the probability of occurrence of major EV serotypes during both periods. Canonical correspondence analysis on relative abundance data was used to test temporal changes in the composition of virus assemblages over the second period. The probability of occurrence of major viruses PV, coxsackieviruses (CVA, CVB), and Echoviruses (E)) significantly changed over time. We found that 1015 isolated PVs were of vaccine origin, called "Sabin-like" (isolates PV1, PV2, PV3). The composition of EV assemblages changed significantly during the second period. We conclude that during the whole period, CVB5, CVB4, and E3 were prominent NPEVS in the SR.

Coxsackievirus B4 sewage-isolate induces pancreatitis after oral infection of mice.

Numerous serotypes which belong to the genus Enterovirus (EV) show variability in their virulence and clinical manifestations. They are also known to undergo changes caused by mutations and recombination during their circulation in the environment and the population. Various EV serotypes are prevalent in groundwater, wastewater and surface waters. Our previous studies showed that oral infection induces pancreatitis depending on specific conditions, such as gravidity, in an outbred murine model. Our aim in the present study was to further explore the pancreatic histopathology in an outbred mouse model following oral infection with clinical isolates from a patient who had aseptic meningitis and an isolate from a treated-sewage sample recovered from the residential area of the patient. The isolates were identified as coxsackievirus B4 (CVB4) in tissue culture. The CVB4 sewage-isolate induced pancreatitis after oral infection. In contrast, pancreatitis was absent following infection with the clinical isolates. Comparison of polyprotein sequences showed that the treated-sewage strains differed from the patient's isolates by 9 and 11 amino acids. We conclude that the isolates of clinical and environmental origin differed in their pathogenic properties and showed genetic variation.

Reproductive health of Roma women in Slovakia.

OBJECTIVE: In most indicators of the way of life, the Roma community is generally different from the majority population and dominant culture. The objective of the study was to describe factors affecting the health of the Roma living in Slovakia, with an emphasis on the sexual and reproductive health of Roma women, and report on the results of analysis of high-risk pregnancies of Roma women in the district of Rimavská Sobota, Slovakia. METHODS: A retrospective study of medical documentation was used. The results were analyzed using the absolute and relative frequencies. Statistical methods were used. RESULTS: A total of 1,256 high-risk pregnancies were analyzed, of which 622 (49.52%) were in Roma women. The average age of Roma respondents was lower by 5 years compared to non-Roma. The age of Roma women at the first pregnancy was statistically significantly lower compared to non-Roma (p < 0.001). The Roma respondents achieved statistically significantly lower levels of education than non-Roma. There was a demonstrably higher number of pregnancies as well as a higher number of artificial and spontaneous abortions per Roma woman. These results were statistically significant. For Roma women, pregnancy began to be risky demonstrably earlier than for non-Roma (p < 0.001). There was a statistically significant difference in attending prenatal counselling. Roma women attended prenatal counselling statistically significantly less frequently than non-Roma (p < 0.001). A significant statistical dependence was found between attending prenatal counselling and the onset of pregnancy problems in Roma women. There was no significant difference in the incidence of other diseases associated with high-risk pregnancy among Roma and non-Roma respondents. CONCLUSION: The findings indicate that Roma women are exposed to health problems in the area of sexual and reproductive health in Slovakia. In the approach to the Roma, it is essential to focus on improving accessibility to health care, prevention, knowledgeableness and effectively preventing, eradicating and strongly penalizing all forms of discrimination in access to health care, especially for Roma women, who are more likely to receive health care.

Reoccurrence of botulinum neurotoxin subtype A3 inducing food-borne botulism, Slovakia, 2015.

A case of food-borne botulism occurred in Slovakia in 2015. Clostridium botulinum type A was isolated from three nearly empty commercial hummus tubes. The product, which was sold in Slovakia and the Czech Republic, was withdrawn from the market and a warning was issued immediately through the European Commission's Rapid Alert System for Food and Feed (RASFF). Further investigation revealed the presence of botulinum neurotoxin (BoNT) subtype BoNT/A3, a very rare subtype implicated in only one previous outbreak (Loch Maree in Scotland, 1922). It is the most divergent subtype of BoNT/A with 15.4% difference at the amino acid level compared with the prototype BoNT/A1. This makes it more prone to evading immunological and PCR-based detection. It is recommended that testing laboratories are advised that this subtype has been associated with food-borne botulism for the second time since the first outbreak almost 100 years ago, and to validate their immunological or PCR-based methods against this divergent subtype.

Should Enteroviruses Be Monitored in Natural Recreational Waters?

Enteroviruses (EVs) infections occur worldwide. Although, infections by these viruses are often asymptomatic and go unnoticed, they can be shed in stool for several weeks. The EVs are associated with sporadic outbreaks and a wide range of clinical symptoms, occasionally accompanied with fatal consequences. Presently in the Slovak Republic (SR) recreational waters are tested only for bacterial indicators. Our aim was to monitor EVs in recreational waters. Water samples were collected during the years 2012-2014 from different recreational natural lakes in Central and West regions of SR. The samples were concentrated by centrifugation using the two-phase separation method recommended by the World Health Organization (WHO) used for EVs surveillance in the treated sewage waste water. Each of the two phases collected from the samples was analysed by polymerase chain reaction for detection of EVs and primary sequencing was done. Our study demonstrated presence of EVs in three localities consecutively for three years, indicating a probability of constant local source of faecal contamination. This is the first monitoring report on the occurrence of EVs in the natural recreational waters in SR.

Factors affecting the success of influenza laboratory diagnosis.

Influenza is one of the most common human infectious diseases, and has profound health and economic consequences. The laboratory diag- nosis of influenza virus infections plays an important role in the global surveillance of influenza. Therefore, there is a growing demand for highly sensitive and rapid methods for detecting influenza. The performance of particular diagnostic methods is affected by various factors. In this study, we assess the effects of patients' age and time to diagnosis on the probability of detecting influenza using four diagnostic methods (virus isolation, rapid test, RT-PCR and real-time RT-PCR). We examined 3,546 samples from central and eastern Slovakia during the influenza seasons from 2005-2006 to 2010-2011. In general, the probability of influenza detection significantly decreased with the time from onset of illness to sample collection (T1) as well as with patients' age (AGE). On the contrary, time from sample collection to delivery (T2) did not play a role in the prob- ability of influenza detection. As judged by odds ratios, the virus isolation method was most sensitive to T1, followed by the rapid test and RT-PCR methods. For the effect of AGE, the rapid test and virus isolation methods were more sensitive than PCR-based methods. The effects of T1 and AGE were independent of each other. Laboratories which participate in inifluenza surveillance should use several methods to enable rapid and accurate influenza A and B virus detection.

A real-time PCR diagnostic method for detection of Naegleria fowleri.

Naegleria fowleri is a free-living amoeba that can cause primary amoebic meningoencephalitis (PAM). While, traditional methods for diagnosing PAM still rely on culture, more current laboratory diagnoses exist based on conventional PCR methods; however, only a few real-time PCR processes have been described as yet. Here, we describe a real-time PCR-based diagnostic method using hybridization fluorescent labelled probes, with a LightCycler instrument and accompanying software (Roche), targeting the Naegleria fowleriMp2Cl5 gene sequence. Using this method, no cross reactivity with other tested epidemiologically relevant prokaryotic and eukaryotic organisms was found. The reaction detection limit was 1 copy of the Mp2Cl5 DNA sequence. This assay could become useful in the rapid laboratory diagnostic assessment of the presence or absence of Naegleria fowleri.

[Monitoring the prevalence of Streptococcus pneumoniae in various groups]. / Monitoring prevalencie Streptococcus pneumoniae v rôznych kolektívoch.

BACKGROUND: The aim of the present study was to monitor the nasopharyngeal presence of Streptococcus pneumoniae in different age groups (especially children) in Banská Bystrica, Slovakia. The purpose of this screening was to determine the prevalence of different serotypes and to follow up the presence of pneumococcus in these children after the vaccination with heptavalent protein-conjugate vaccine. A contribution of molecular biology techniques was the detection of S. pneumoniae DNA by PCR and also the typisation and comparison of pneumococcal strains by pulsed-field gel electrophoresis. METHODS: S. pneumoniae in nasopharyngeal swabs was detected by cultivation on blood agar plates. Serotypisation was performed by standard Quellung reaction. The commercial diagnostic kit was used for PCR detection of S. pneumoniae DNA. Pulsed-field electrophoresis was performed by modified scheme according to literature. RESULTS: The incidence of pneumococcus is decreasing and less significant with the increasing age. Among youngest children is relatively high prevalence of pneumococci and the relatedness of isolated strains is high as well. After the vaccination, the less invasive serotypes were detected, although the overall incidence of S. pneumoniae was similar. CONCLUSIONS: The monitoring of S. pneumoniae in population is important according to variability of this bacteria with respect to possible changes in pneumococcal types as a consequence of vaccination.

Polymerase chain reaction (PCR)--an efficient tool for diagnosis of influenza and acute respiratory illnesses during influenza season.

Many severe diseases of the respiratory tract lead to hospitalisation. These diseases are often caused by viral infections and may cause increased mortality. The most common viral pathogens involved in these cases, which are also associated with significant morbidity and mortality during the influenza seasons are influenza viruses. Rapid differential diagnosis of influenza viruses is therefore of great importance. Classical diagnosis of these viruses involves virus cultures. Of the rapid diagnostic methodologies which have been developed are RT-PCR, multiplex PCR, real-time PCR. In the present study we have monitored clinical samples from patients of different age groups from selected regions in Slovakia and compared the effectiveness of the classical and molecular biological diagnostic methods. The molecular biological methods proved to be rapid, accurate and effective. Application of these techniques in diagnosis of the respiratory illnesses should help in the prevention, therapy and disease control.

First confirmation of Bordetella pertussis occurrence in Slovakia by using real-time PCR.

By application of the real-time PCR we manage to confirm the diagnosis and occurrence of a disease, which is caused by Bordetella pertussis-pertussis. Using this method we have proven the presence of DNA of Bordetella pertussis in the biological materials (nasopharyngeal swabs). The presence of IS481 genome sequence of Bordetella pertussis was confirmed. This method of detection of pathogens seems to be very rapid, simple, and specific. In the case of adequate technical laboratory equipment it may become very suitable and important supporter in explanation and confirmation of the occurrence of bacterial infections.