Renata Kissova, MSc., PhD is a Virologist currently heads the Virology Laboratory of the Regional Authority of Public Health in Banska Bystrica, Slovak Republic. Her work is focused mainly on the surveillance and diagnosis of enteroviruses and influenza viruses, especially virus isolations in cell cultures techniques. She received her MSc. Microbiology degree from the Safarik University in Kosice and the Slovak Medical University in Bratislava, Slovakia. She got her PhD. at the Faculty of Public Health of Slovak medical University in Bratislava, Slovakia. Since 2013 she has been teaching at the Slovak Medical University, Faculty of Health in Banska Bystrica
We present the monitoring programme of sewage water according to the World Health Organization (WHO) strategy for polio eradication in the Slovak Republic (SR). Polioviruses (PV) and non-polio enteroviruses (NPEV). \r\nWe cover the period from 2001 to 2016, which ranges from before and after the change in polio vaccination strategy, in the Slovak Republic. Samples from the sewage treatment plants of 48 localities from Western, Central and Eastern regions were tested. We have compared the circulation pattern of PVs and NPEVs in the SR, as a component of the PV surveillance program. WHO standard procedures were followed for virus isolations and identiﬁcations. 870 human enteroviruses (EVs) were detected: 357 (41%) coxsackie B viruses (CBV), 309 (36%) echoviruses, 76 (9%) NPEV not-typed and 114 (13%) Sabin-like PVs (PV1, 2, 3) including vaccine-derived poliovirus (VDPV) isolates. The percentage of PV isolates fell from 66% to 30% during 2001–2005 and thereafter to zero. CBV5, echoviruses 3 and 11 were the NPEVs endemic during the study period. In conclusion we have shown that the changes in the vaccination program of oral polio vaccine towards inactivated polio vaccine in the year 2005 changed the balance of circulating serotypes from Sabin-like PV and vaccine derived PV towards the NPEV. In September 2015, wild PV type 2 was officially declared eradicated. According to the WHO Global Action Plan to minimize poliovirus facility-associated risk after type-specific eradication of wild polioviruses, Slovak republic disposed all material containing PV2.\r\n
Brigita Benkőová, MSc, is a Virologist currently working at the Enterovirus Laboratory and the National Reference Centre for Identification of Enteroviruses at the Medical Faculty of the Slovak Medical University in Bratislava, Slovakia. She received her BSc. and MSc. Virology degree at Comenius University in Bratislava, Faculty of Natural Sciences. Her Bachelor thesis contains a literature review of neonatal enteroviral infections, their pathogenesis and clinical manifestation. In her MSc research thesis was focused on adaptive and innate immune response caused by coxsackievirus infection.\r\n\r\n
Coxsackieviruses (CV) are small positive-stranded RNA viruses, important human pathogens that cause both acute and chronic diseases. These viruses induce lytic infections; their cytopathic effect (CPE) includes morphological changes and destruction of the host cell monolayer in vitro. Their viral RNA can remain persistent for prolonged times in cells and organs post infection. Literature reports show moderate replication of enteroviruses in NIH 3T3 cells. Our aim was to study the replication of coxsackieviruses in NIH 3T3 clones with resistance to Puromycin alone or along with Blastomycin, in combination with presence of a truncated variant of the Dicer ribonuclease. Four genetically modified clones of NIH 3T3 cells were infected with CVB3 (Nancy strain). This virus was previously passaged in Vero cells with a titer of 106.75 TCID50. Cells were infected with a multiplicity of infection of 0.1. We checked the cells for presence of viral replication (CPE) and viral RNA by the reverse transcriptase polymerase chain reaction (PCR) and Nested-PCR. We also made an attempt to adapt the virus to the cells by blind passages in the NIH 3T3 cells. Morphological changes were absent in the infected cells as compared to the mock-infected control of the NIH 3T3 cell lines. After third and fourth passages of the virus in the same cell line, rounding of cells was observed but this effect did not increase on further passages of the virus. When checked for presence of viral RNA, the cultures were found to be positive irrelevant of the rounding effect. Considering the viral kinetics we hypothesize a slow viral replication in the first phase of infection in these cells which might allow a time frame for the induction of cytokines, resulting in further slowing down the virus replication and spread of infection between cells and ensure cell survival and leading to persistence of viral RNA.