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In the context of the near-global eradication of wild poliovirus, the significance of non-polio enteroviruses (NPEVs) in causing acute flaccid paralysis (AFP) and their impact on public health has gained increased attention. This research, conducted from 2001 to 2021, examined stool samples from 1597 children under 15 years in São Paulo, Brazil, through the AFP/Poliomyelitis Surveillance Program, detecting NPEVs in 6.9% of cases. Among the 100 NPEV-positive strains analyzed, 90 were genotyped through genomic sequencing of the partial VP1 region, revealing a predominance of EV-B species (58.9%), followed by EV-A (27.8%) and EV-C (13.3%). This study identified 31 unique NPEV types, including EV-A71, CVB2, and E11, as the most prevalent, along with the first documented occurrence of CVA19 in Brazil. These findings emphasize the importance of NPEV genotyping in distinguishing AFP from poliomyelitis, enhancing understanding of these viruses' epidemiology. Moreover, it ensures that AFP cases are correctly classified, contributing to the effective surveillance and eradication efforts for poliomyelitis. [ABSTRACT FROM AUTHOR]

Purpose: Recently, cases of serious illness in newborns infected with Echovirus 11 have been reported in Europe, including Italy. Here, we report the case of a newborn diagnosed with disseminated Echovirus 11 infection, which occurred in October 2023 in the Province of Bolzano, Italy. Methods: A molecular screening, by Real-Time RT-PCR, was employed to analyse the cerebrospinal fluid, blood and stool samples, and nasal swabs. The entire viral genome was sequenced using both Illumina and Nanopore technologies. Results: The patient was admitted to hospital due to fever. Molecular testing revealed the presence of enterovirus RNA. Typing confirmed the presence of Echovirus 11. The patient was initially treated with antibiotic therapy and, following the diagnosis of enterovirus infection, also with human immunoglobulins. Over the following days, the patient remained afebrile, with decreasing inflammation indices and in excellent general condition. Genomic and phylogenetic characterization suggested that the strain was similar to strains from severe cases reported in Europe. Conclusions: Despite the low overall risk for the neonatal population in Europe, recent cases of Echovirus 11 have highlighted the importance of surveillance and complete genome sequencing is fundamental to understanding the phylogenetic relationships of Echovirus 11 variants. [ABSTRACT FROM AUTHOR]

Introduction. Enteroviruses (EV) are characterized by: species and type diversity, polymorphism of clinical manifestations, a tendency to epidemic spread, and are often the cause of disease outbreaks, which determines the relevance of monitoring EV strains in various clinical forms of infection, including in conditions of anti-epidemic measures. The aim of the study: to characterize the prevalence and diversity of non-polio enteroviruses (NPEV) types in children with acute intestinal infection (AII) in the period 2018–2023, including the COVID-19 pandemic. Materials and methods. The RT-PCR method was used to study 7302 samples of feces from children hospitalized with a diagnosis of AII in the infectious diseases hospital of Nizhny Novgorod. Genotyping of EV strains was carried out using fragment Sanger sequencing of the genome region encoding capsid protein 1 (VP1) and the online resource BLAST. Results. EVs were found in 5.0 ± 0.3% (1.7–7.8%), both in mono- and mixed infections with other enteric viruses. The long-term dynamics of the frequency of EV detection and the incidence of EV infection in children in the Nizhny Novgorod region was characterized by a sharp decrease in indicators in 2020 against the backdrop of the introduction of anti-epidemic measures. When genotyping 299 strains, 41 types of NPEV of 4 species were identified. The spectrum included the main pathogens of exanthema and neuroinfections and rare types found in “minor” or intestinal forms of infection. During the study period, a redistribution of NPEV species was established. Before the pandemic, the ratio of Enterovirus A : Enterovirus B : Enterovirus C species was as follows — 41.0 : 46.7 : 12.3%; during the 2020 pandemic season the ratio was 0.0 : 37.5 : 62.5%; after the lifting of restrictive measures — 47 : 29 : 23%, which may be due to the different effectiveness of the restrictive measures on the mechanisms of transmission of EVs of different types. Conclusion. The genetic diversity of NPEVs detected in children with AII complements information on the typical composition of the territorial enterovirus population. In children with AII, when the airborne transmission of SARS-CoV-2 was blocked, there was a decrease in the frequency of detection of viruses of the Enterovirus B type, the absence of detection of Enterovirus A and the constant presence of Enterovirus C.

Objectives: Several factors can cause acute flaccid paralysis cases including non-polio enteroviruses. In Senegal, few studies on non-polio enteroviruses (NPEV) have been performed. Methods: Our study assess the molecular epidemiology of non-polio enteroviruses in Senegal from 2013 to 2021 through the previously existing programs for surveillance of polioviruses. Results: A total of 3815 stool samples and 281 sewage samples were collected. After virus isolation by cell culture, non-polio enteroviruses-positive isolates were confirmed by reverse transcriptase-quantitative polymerase chain reaction. Following this detection, the positive samples were subjected to molecular characterization. Our data showed that 15.22% and 52.66% were positive in cell culture for non-polio enteroviruses in acute flaccid paralysis surveillance and environmental surveillance, respectively. These non-polio enteroviruses-positive isolates were detected all year round but tend to unequal peaks of circulation, and the age group 0-5 years was more vulnerable to infection (84.4%). Genetic characterization revealed the circulation of enteroviruses species infecting humans (Enterovirus A – Enterovirus D): Enterovirus A (29.2%) and Enterovirus B (63.1%) isolates from both the acute flaccid paralysis surveillance and environmental surveillance while Enterovirus C (5.3%) and Enterovirus D (2.4%) were only isolated from the acute flaccid paralysis surveillance. However, the highly prevalent Enterovirus B species from the acute flaccid paralysis surveillance included echovirus 7 and echovirus 13, whereas coxsackievirus A6 was the predominant species from the environmental surveillance. Conclusion: This first 8-year period study of NPEV in Senegal showed that NPEV represent important viral etiologies associated with acute flaccid paralysis cases and circulating in environmental surveillance in Senegal and highlighted the need to promote effective long-term strategies for monitoring of non-polio enteroviruses infections.

Enteroviruses are one of the most common causative agents of infectious diseases of the central nervous system. They are characterized by genetic variability, the ability to infect a wide range of cells, including brain microglial cells and astrocytes, and persist in the central nervous system tissue, causing delayed and chronic diseases. The review presents data on the basis of neurovirulence of non-polio enteroviruses and the most common pathogens causing enteroviral neuroinfections.

More than 100 types of non-polio enteroviruses (NPEVs) are ubiquitous in the human population and cause a variety of symptoms ranging from very mild to meningitis and acute flaccid paralysis (AFP). Much of the information regarding diverse pathogenic properties of NPEVs comes from the surveillance of poliovirus, which also yields NPEV. The analysis of 265 NPEV isolations from 10,433 AFP cases over 24 years of surveillance and more than 2500 NPEV findings in patients without severe neurological lesions suggests that types EV-A71, E13, and E25 were significantly associated with AFP. EV-A71 was also significantly more common among AFP patients who had fever at the onset and residual paralysis compared to all AFP cases. In addition, a significant disparity was noticed between types that were common in humans (CV-A2, CVA9, EV-A71, E9, and E30) or in sewage (CVA7, E3, E7, E11, E12, and E19). Therefore, there is significant evidence of non-polio viruses being implicated in severe neurological lesions, but further multicenter studies using uniform methodology are needed for a definitive conclusion. [ABSTRACT FROM AUTHOR]

Human non-polio enteroviruses (NPEVs) are the etiological agents involved in most cases of hand-foot-and-mouth disease (HFMD), herpangina and aseptic meningitis. Information on the epidemiology profiles of NPEV in the Ural Federal District and Western Siberia is very limited, with no published data available. The aim of this study is to describe NPEV incidence in the Ural Federal District and Western Siberia among patients with different forms of non-polio enterovirus infections (NPEVIs) during 2022, stratified by age and clinical manifestations. A total of 265 samples that tested positive for NPEV using a polymerase chain reaction (PCR) were genotyped by semi-nested PCR for the VP1 gene. The results showed that 21 genotypes were identified among patients in this study. CVA6 was the most common genotype for HFMD. CVA6, along with CVA10, accounted for the majority of herpangina cases, while CVA9 was implicated in most meningitis cases. Sequence and phylogenetic analysis showed that nearly all of the CVA6 strains identified in this study displayed a close genetic relationship to strains identified in other cities in Russia and strains from China. NPEV surveillance allows for monitoring the circulation of clinically relevant genotypes, resulting in continuous data about NPEV epidemiology. This is important for improving case prevention, diagnosis and guiding clinical management. [ABSTRACT FROM AUTHOR]

Background and Aim: During the COVID-19 pandemic public health measures and protocols such as regular hand washing and wearing a face mask were recommended at the national level. The implementation of these health protocols reduced the prevalence of respiratory diseases, but no study has been conducted to investigate the impact of the implementation of these protocols on the spread of viruses that are transmitted through fecal-oral route. Considering that non-polio enteroviruses (NPEVs) are mainly transmitted through fecal-oral route, this study was conducted to determine the prevalence of non-polio enteroviruses in patients with acute flaccid paralysis (AFP) in Iran before and during the COVID-19 pandemic. Materials and Methods: To detect non-polio enteroviruses stool samples of AFP cases received by Iran National Polio Laboratory in 2019 (before the COVID-19 pandemic) and 2021 (during the COVID-19 pandemic) were tested. To isolate NPEVs by cell culture, the WHO standard protocol was used, and the TaqMan One-Step Real Time PCR was used for molecular testing. Results: In 2019, a sample of 21 out of 1070 patients and a sample of 12 out of 100 patients were found to be positive for non-polio enterovirus by cell culture and the molecular method, respectively; these ratios decreased to10 out of 678 and 3 out of 100 in 2021. Conclusion: Based on the findings of this study, it can be concluded that the observance of personal hygiene and public health protocols during the COVID-19 epidemic have, in addition to the general reduction of respiratory transmission of viruses, also affected the faecal-oral transmission of viruses. [ABSTRACT FROM AUTHOR]

Non-polio enteroviruses (NPEVs) are ubiquitous and are one of the main causative agents of viral infections in children. NPEVs most commonly infect newborns and young children, due to their lack of antibodies. In children, clinical manifestations can range from acute febrile illness to severe complications that require hospitalization and lead in some cases to disability or death. NPEV infections can have severe consequences, such as polio-like diseases, serous meningitis, meningoencephalitis, myocarditis, etc. The most promising strategy for preventing such diseases is vaccination. No less than 53 types of NPEVs have been found to circulate in Russia. However, of epidemic importance are the causative agents of exanthemic forms of the disease, aseptic meningitis and myocarditis. At the same time, the frequency of NPEV detection in the constituent entities of the Russian Federation is characterized by uneven distribution and seasonal upsurges. The review discusses the epidemic significance of different types of enteroviruses, including those relevant to the Russian Federation, as well as current technologies used to create enterovirus vaccines for the prevention of serious diseases.

ABSTRACT Continuous surveillance of enteroviruses (EVs) in urban domestic sewage can timely reflect the circulation of EVs in the environment and crowds, and play a predictive and early warning role in EV-related diseases. To better understand the long-term epidemiological trends of circulating EVs and EV-related diseases, we conducted a 9-year (2013 to 2021) surveillance study of non-polio EVs (NPEVs) in urban sewage in Guangzhou city, China. After concentrating and isolating the viruses from the sewage samples, NPEVs were detected and molecular typing was performed. Twenty-one different NPEV serotypes were identified. The most isolated EVs were echovirus 11 (E11), followed by coxsackievirus (CV) B5, E6, and CVB3. EV species B prevailed in sewage samples, but variations in the annual frequency of different serotypes were also observed in different seasons, due to spatial and temporal factors. E11 and E6 were detected continuously before 2017, and the number of isolates was relatively stable during the surveillance period. However, after their explosive growth in 2018 and 2019, their numbers suddenly decreased significantly. CVB3 and CVB5 had alternating trends; CVB5 was most frequently detected in 2013 to 2014 and 2017 to 2018, while CVB3 was most frequently detected in 2015 to 2016 and 2020 to 2021. Phylogenetic analysis showed that at least two different transmission chains of CVB3 and CVB5 were prevalent in Guangzhou City. Our results show that in the absence of a comprehensive and systematic EV-related disease surveillance system in China, environmental surveillance is a powerful and effective tool to strengthen and further investigate the invisible transmission of EVs in the population. IMPORTANCE This study surveilled urban sewage samples from north China for 9 years to monitor enteroviruses. Samples were collected, processed, and viral identification and molecular typing were performed. We detected 21 different non-polio enteroviruses (NPEVs) with yearly variations in prevalence and peak seasons. In addition, this study is very important for understanding the epidemiology of EVs during the COVID-19 pandemic, as the detection frequency and serotypes of EVs in sewage changed considerably around 2020. We believe that our study makes a significant contribution to the literature because our results strongly suggest that environmental surveillance is an exceptionally important tool, which can be employed to detect and monitor organisms of public health concern, which would otherwise be missed and under-reported by case-based surveillance systems alone.

Human non-polio enteroviruses (NPEVs) are the etiological agents involved in most cases of hand-foot-and-mouth disease (HFMD), herpangina and aseptic meningitis. Information on the epidemiology profiles of NPEV in the Ural Federal District and Western Siberia is very limited, with no published data available. The aim of this study is to describe NPEV incidence in the Ural Federal District and Western Siberia among patients with different forms of non-polio enterovirus infections (NPEVIs) during 2022, stratified by age and clinical manifestations. A total of 265 samples that tested positive for NPEV using a polymerase chain reaction (PCR) were genotyped by semi-nested PCR for the VP1 gene. The results showed that 21 genotypes were identified among patients in this study. CVA6 was the most common genotype for HFMD. CVA6, along with CVA10, accounted for the majority of herpangina cases, while CVA9 was implicated in most meningitis cases. Sequence and phylogenetic analysis showed that nearly all of the CVA6 strains identified in this study displayed a close genetic relationship to strains identified in other cities in Russia and strains from China. NPEV surveillance allows for monitoring the circulation of clinically relevant genotypes, resulting in continuous data about NPEV epidemiology. This is important for improving case prevention, diagnosis and guiding clinical management.

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background. Non-polio enteroviruses (EVs) and human parechoviruses (PeVs) cause a wide range of human infections. Limited data on their true disease burden exist as standardized European-wide surveillance is lacking. Aims. Our aim is to estimate the disease burden of EV and PeV infections in Europe via establishment of standardized surveillance for hand, foot and mouth disease (HFMD) and respiratory and neurological infections caused by these viruses. We will also assess the sensitivity of assays implemented in the network of participating laboratories so that all EV and PeV types are adequately detected. Plan. The European Non-Polio Enterovirus Network (ENPEN) has developed standardized protocols for a prospective, multi-center and cross-sectional hospital-based pilot study. Protocols include guidance for diagnosis, case definition, detection, characterization and reporting of EV and PeV infections associated with HFMD and respiratory and neurological diseases. Over 30 sites from 17 European countries have already registered to this one pilot study, likely to be commenced in 2022. Benefits. This surveillance will allow European-wide comparison of data on EV and PeV infection. These data will also be used to determine the burden of EV and PeV infections, which is needed to guide the further prevention measures and policies. [ABSTRACT FROM AUTHOR]

Objective: To investigate the prevalence of viral myocarditis worldwide. Methods: We conducted a systematic search for the prevalence of the most common viruses in myocarditis and 75 studies were included for statistical analysis of the prevalence of adenovirus, hepatitis C virus, cytomegalovirus, Ebola virus, human herpesvirus 6, influenza virus, parvovirus, and non-polio enteroviruses. Results: The highest prevalence was related to B19 (25.0%) and non-polio enteroviruses (18%). The prevalence of human herpesvirus 6, cytomegalovirus, and Ebola virus was12.8%, 5.5%, and 3.1%, respectively. Hepatitis C virus accounted for 6.1% of the disease, the adenoviruses contributed to 5.2% of viral myocarditis. The lowest incidence was related to the influenza virus with 2.0%. Conclusions: Treatment of myocarditis is still problematic and may depend on the etiologic diagnosis. So it is important to know the commonly occurring viral factors in myocarditis and timely diagnosis and treatment are also imperative.

The article describes two intrafamilial foci of enterovirus infection (EVI), caused by ECHO-3 and ECHO-6 viruses. In the first group, 6 children aged from 3 months to 4 years were found (average age — 18.0 ± 4.3 months), 3 boys (50 %) and 3 girls (50 %) with various clinical manifestations of EVI associated between themselves epidemiologically (siblings of one fami­ly, who used water from one draw-well and had close household contact). ECHO-3 virus was detected in the clinical material of the examined children. The second outbreak involved two children from the same family, who simultaneously suffered from aseptic meningitis in 2017. Cerebrospinal fluid of both children was examined by reverse transcription polymerase chain reaction (RT-PCR) (Republican Research and Practical Center for Epidemiology and Microbiology, Minsk), RNA EV was detected in both samples. After RNA sequencing, ECHO 6 was detected in both samples. The verification in both cases was performed by various clinical and laboratory methods (general clinical methods, enzyme-linked immunosorbent assay with detection of antibo­dies to EV and antigens of EV in feces, RT-PCR with subsequent EV RNA sequencing and determination of virus serotype). In two cases of the disease, the diagnosis of EVI was made epidemiologically without laboratory detection of RNA EV. PCR with the determination of EV RNA in feces and cerebrospinal fluid showed the greatest effectiveness. In the described outbreaks, RNA of enteroviruses was not detected in the blood in any sample.

In addition to polioviruses, non-polio enteroviruses (NPEVs) are frequently isolated from patients with acute flaccid paralysis (AFP) worldwide. In polio-free countries, there have been expectations that with disappearing wild poliovirus from the community, the rate of AFP would decrease, but the increasing number of AFP cases proved this notion to be wrong. There are speculations that NPEVs might be the cause of increasing AFP rate. The aim of this study was to investigate frequency, genetic diversity, circulation patterns of NPEVs isolated from AFP cases in Iran from 2015 to 2018. Fifty-three NPEVs were isolated from stool specimens of AFP cases during four years of AFP surveillance. Nested PCR and VP1 sequencing revealed 20 NPEV types in which Echovirus 3 (13.2%), Echovirus 6 (13.2%), Echovirus 7 (7.5%), Echovirus 13 (7.5%) and Echovirus 21 (7.5%) were the most frequent. Coxsackie B viruses were isolated for the first time in AFP cases in Iran. The phylogenetic analysis of Echovirus 3 and Echovirus 6 revealed that Iranian echovirus strains belonged to the same cluster, indicating these viruses have been circulating in Iran for a long time. Compared to global Echovirus 3 and Echovirus 6 references, Echovirus 3 and Echovirus 6 strains detected in this study were closely related to Indian and Malaysia strains, respectively. The results of this study demonstrated a wide variety of NPEV types in Iranian patients, some of which had not been reported in previous studies. Moreover, this study highlights the need for NPEV surveillance in AFP cases. [ABSTRACT FROM AUTHOR]

Background: Human non-polio enteroviruses (NPEVs) have been associated with certain life-threatening disorders in children. However, there is paucity of NPEV infection data in most developing countries. This study determined the 3-year prevalence of non-polio enteroviruses (NPEVs) among children in some Northern States of Nigeria. Materials and Methods: Duplicate stool samples were collected from 27778 children =15 years. These samples were processed and analyzed for characteristic NPEVs cytopathic effects (CPE) on L20B and RD cell lines. Tests were considered positive if the duplicate samples produced distinct CPE on both cell lines. Results: Of the 27778 samples processed, 3991 (14.4%) NPEVs were isolated. Participants of the male gender (14.5%) within the age range of 0-5 years (14.7%) from Yobe state (15.3%) whose samples were received in the month of June (22.2%) and in the year 2015 (18.8%) had the highest prevalence of NPEVs. June had significant risk factors of NPEVs (p=0.001, OR=1.95 [95%CI: 1.60-2.34]). However, there was no significant association between age, sex and location of sample collection with the prevalence of NPEV (p=0.05) Conclusions: This study revealed a relatively high prevalence of NPEVs among the study population. This calls for the need for government implementation of consistently improved water, food and environmental hygiene. [ABSTRACT FROM AUTHOR]

Surveillance for acute flaccid paralysis syndrome (AFP) in children under 15 is the backbone of the Global Polio Eradication Initiative. Laboratory examination of stool samples from AFP cases allows the detection of, along with polioviruses, a variety of non-polio enteroviruses (NPEV). The etiological significance of these viruses in the occurrence of AFP cases has been definitively established only for enteroviruses A71 and D68. Enterovirus Coxsackie A2 (CVA2) is most often associated with vesicular pharyngitis and hand, foot and mouth disease. Among 7280 AFP cases registered in Russia over 20 years (2001–2020), CVA2 was isolated only from five cases. However, these included three children aged 3 to 4 years, without overt immune deficiency, immunized with 4–5 doses of poliovirus vaccine in accordance with the National Vaccination Schedule. The disease resulted in persistent residual paralysis. Clinical and laboratory data corresponded to poliomyelitis developing during poliovirus infection. These findings are compatible with CVA2 being the cause of AFP. Molecular analysis of CVA2 from these patients and a number of AFP cases in other countries did not reveal association with a specific phylogenetic group, suggesting that virus genetics is unlikely to explain the pathogenic profile. The overall results highlight the value of AFP surveillance not just for polio control but for studies of uncommon AFP agents.

Here we analyzed six years of acute flaccid paralysis (AFP) surveillance, from 2015 to 2020, of 10 countries linked to the WHO Regional Reference Laboratory, at the Istituto Superiore di Sanità, Italy. The analysis also comprises the polio vaccine coverage available (2015–2019) and enterovirus (EV) identification and typing data. Centralized Information System for Infectious Diseases and Laboratory Data Management System databases were used to obtain data on AFP indicators and laboratory performance and countries’ vaccine coverage from 2015 to 2019. EV isolation, identification, and typing were performed by each country according to WHO protocols. Overall, a general AFP underreporting was observed. Non-Polio Enterovirus (NPEV) typing showed a high heterogeneity: over the years, several genotypes of coxsackievirus and echovirus have been identified. The polio vaccine coverage, for the data available, differs among countries. This evaluation allows for the collection, for the first time, of data from the countries of the Balkan area regarding AFP surveillance and polio vaccine coverage. The need, for some countries, to enhance the surveillance systems and to promote the polio vaccine uptake, in order to maintain the polio-free status, is evident.

Background. Non-polio enteroviruses (EVs) and human parechoviruses (PeVs) cause a wide range of human infections. Limited data on their true disease burden exist as standardized European-wide surveillance is lacking. Aims. Our aim is to estimate the disease burden of EV and PeV infections in Europe via establishment of standardized surveillance for hand, foot and mouth disease (HFMD) and respiratory and neurological infections caused by these viruses. We will also assess the sensitivity of assays implemented in the network of participating laboratories so that all EV and PeV types are adequately detected. Plan. The European Non-Polio Enterovirus Network (ENPEN) has developed standardized protocols for a prospective, multi-center and cross-sectional hospital-based pilot study. Protocols include guidance for diagnosis, case definition, detection, characterization and reporting of EV and PeV infections associated with HFMD and respiratory and neurological diseases. Over 30 sites from 17 European countries have already registered to this one pilot study, likely to be commenced in 2022. Benefits. This surveillance will allow European-wide comparison of data on EV and PeV infection. These data will also be used to determine the burden of EV and PeV infections, which is needed to guide the further prevention measures and policies.

Abstract Background The need to investigate the contribution of non-polio enteroviruses to acute flaccid paralysis (AFP) cannot be over emphasized as we move towards a poliovirus free world. Hence, we aim to identify non-polio enteroviruses recovered from the faeces of children diagnosed with AFP in Nigeria. Methods Ninety-six isolates, (95 unidentified and one previously confirmed Sabin poliovirus 3) recovered on RD cell culture from the stool of children

Hereby, the partial Viral Protein 1 sequences of Coxsackievirus B5 (CV-B5) from sewage samples, collected in Italy from 2016 to 2017, were compared with those available in GenBank from clinical samples. Phylogenetic analysis highlighted: (I) the predominant circulation of CV-B5 genogroup B in Italy, and (II) the presence of two new sub-genogroups. [ABSTRACT FROM AUTHOR]

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Abstract Non-Polio EnteroViruses (NPEV) are one of the known causative agents of Acute Flaccid Paralysis (AFP). In the present study, we identified, sequenced and characterized the complete genome of sixty-five Coxsackievirus-A10, an NPEV. These were isolated from stool specimens of AFP cases from Bihar, Karnataka, Kerala, and Uttar Pradesh (UP) states of India. Evolutionary analysis of complete genome (7420 nucleotides) and VP1 gene (894 nucleotides) demonstrates that there are four different intra-typic strains circulating in India which were dissimilar to Chinese strains. First intratypic strain circulating in UP, Bihar, and Karnataka; second in UP and Karnataka; third in UP and Bihar and; fourth was restricted only to Kerala state. The divergence of Kerala strain with respect to all other circulating strain of UP, Bihar and Karnataka states in India is 24%, 24.9%, and 24.4% respectively. Recombinations were observed between few of these strains which might be one of the factors of the observed intra-typic diversity. Article summary line We report the identification, characterization and phylogenetic analysis of sixty-five Non-Polio Enterovirus (NPEV) isolates, performed during the year 2009–17, causing acute flaccid paralysis in pediatric cases with their divergences and recombinations from four states of India. Highlights • Percentage of Non-Polio Enteroviruses (NPEV) detections are increasing in India. • There are multiple NPEVs causing Acute Flaccid Paralysis that varies from region to region. • This study identifies Coxsackievirus A10 from stool samples of AFP cases from Bihar, Karnataka, Kerala, and Uttar Pradesh states of India. • Four different intra-typic strains were found to be circulating in India which was dissimilar to Chinese strains. [ABSTRACT FROM AUTHOR]

Introduction : Viral meningitis is an inflammation of the leptomeninges as a manifestation of central nervous system (CNS) infection, and more than 85% of viral meningitis cases are caused by non-polio enteroviruses. Molecular methods such as polymerase chain reaction (PCR) are increasingly used to detect and type non-polio enteroviruses. Here, we describe the impact of these viruses on aseptic meningitis in young children in Iran. Methods: Cerebrospinal fluids were collected from 366 children under 8 years old with primary clinical diagnosis of aseptic meningitis. Real-Time RT-PCR and PCR were performed using specific primers for each human enterovirus (HEV), human parechovirus (HPeV), and enterovirus 71 (EV71). Results: Positive results were obtained for HEV, HPeV, and EV71 in 118 (32.2%), 154 (42%), and 94 (25.7%) patients, respectively. HPeV detection rate was higher in both male and female patients with no statistically significant difference. Conclusion: Using molecular methods, such as PCR as routine laboratory diagnostic tests for HEVs will result in better understanding of disease burden of these viruses, especially in central nervous system involvements, and will consequently reduce hospitalization and use of antibiotic, which often lead to other complications. J Med Microbiol Infec Dis, 2014, 2 (2): 5 pages.

Background: The need to investigate the contribution of non-polio enteroviruses to acute flaccid paralysis (AFP) cannot be over emphasized as we move towards a poliovirus free world. Hence, we aim to identify non-polio enteroviruses recovered from the faeces of children diagnosed with AFP in Nigeria. Methods: Ninety-six isolates, (95 unidentified and one previously confirmed Sabin poliovirus 3) recovered on RD cell culture from the stool of children <15 years old diagnosed with AFP in 2014 were analyzed. All isolates were subjected to RNA extraction, cDNA synthesis and three different PCR reactions (one panenterovirus 5'-UTR and two different VP1 amplification assays). VP1 amplicons were then sequenced and isolates identified. Results: 92.71% (89/96) of the isolates were detected by at least one of the three assays as an enterovirus. Precisely, 79.17% (76/96), 6.25% (6/96), 7.30% (7/96) and 7.30% (7/96) of the isolates were positive for both, positive and negative, negative and positive, as well as negative for both the 5'-UTR and VP1 assays, respectively. In this study, sixty-nine (69) of the 83 VP1 amplicons sequenced were identified as 27 different enterovirus types. The most commonly detected were CV-B3 (10 isolates) and EV-B75 (5 isolates). Specifically, one, twenty-four and two of the enterovirus types identified in this study belong to EV-A, EV-B and EV-C respectively. Conclusions: This study reports the circulating strains of 27 non-polio enterovirus types in Nigerian children with AFP in 2014 and Nigerian strains of CV-B2, CV-B4, E17, EV-B80, EV-B73, EV-B97, EV-B93, EV-C99 and EV-A120 were reported for the first time. Furthermore, it shows that being positive for the 5'-UTR assay should not be the basis for subjecting isolates to the VP1 assays. [ABSTRACT FROM AUTHOR]

Background: In some countries, though wild poliovirus isolation from clinical specimens has not been done, evidences of silent circulation of wild virus in sewage system have been reported. On the other hand, based on WHO recommendations, if there is any probability of wild virus entrance from endemic countries, environmental surveillance is advised as a completion to Acute Flaccid Paralysis (AFP) surveillance. Methods: AFP and environmental surveillance were performed simultaneously in Sistan and Balouchestan province to confirm wild poliovirus eradication. To perform AFP surveillance, 21 stool specimens of AFP suspected cases were collected in a one year period and evaluated using L20B, RD and Hep-2 cell lines. To perform environmental surveillance, 86 samples from urban and hospital sewage disposal systems were collected. Using direct, Pellet and Two-phase methods for virus concentration, Enterovirus existence was investigated in above-mentioned cell lines. Results: Non-polio enteroviruses and polioviruses were isolated from 53.49% and 20.93% of sewage specimens, respectively. Sabin-Like polioviruses were isolated from 9.52% of AFP cases, and non-polio enteroviruses from 14.28% of them. Poliovirus type 2 was not isolated form stool specimens and there was no Poliovirus type 1 isolation from sewage specimens, either. Conclusion: The environmental surveillance as a completion to AFP in Sistan and Balouchestan Province in Iran confirms poliovirus eradication.

Introduction: Poliovirus infections have been established to be in circulation in the remaining three polio-endemic nations. These pathogens have been associated with several chronic diseases, particularly acute flaccid paralysis of children. This study sought to ascertain whether polioviruses are silently shed by apparently healthy schoolchildren in Bauchi, Katagum, and Misau local government areas of Bauchi state, Nigeria. Methodology: This was a cross-sectional prospective study that involved 200 stool samples collected from apparently healthy schoolchildren. All samples were processed and inoculated onto rhabdomyosarcoma (RD) and L20B cell-lines. Inoculated cell lines were monitored for cytopathic effects (CPEs) for 10 days with one subculture after first 5 days. Results: None of the samples came down with CPEs on L20B, and thus all samples were negative for poliovirus; however, three were positive for non-polio enteroviruses (NPEVs) on RD and not on the L20B cell line: one coxsackie B virus from a seven-year-old male, and two others were untypeable isolates, one each from a male and a female child. The coxsackie B virus was identified by microneutralization test using polyclonal sera as described by the World Health Organization. Conclusions: Findings from this investigation indicate the absence of polioviruses in the children studied. This is an indication of good polio immunization coverage in these communities. However, more intensive and periodic surveillance is required to confirm the presence or exclude the absence of polioviruses in these communities and other parts of Nigeria. [ABSTRACT FROM AUTHOR]

Background: In the global polio eradication initiative acute flaccid paralysis cases are followed up two to three months after onset of paralysis to assess recovery of the children. In Zambia AFP cases are followed up regularly but there is no documentation of the clinical and laboratory findings. The purpose of this paper is to document the support WHO country office offers to the follow up of the AFP cases in Zambia to identify gaps which the WHO Country office could address. Methods: This study used secondary data from a case control study design, conducted and supported by WHO country office, which was aimed at assessing the association of NPEVs with AFP. Particularly this study aims at assessing the presence or absence of residual paralysis and the laboratory findings of the affected children. Results: A total of 93 cases of, which over 55% were males were included in this study. Majority of the cases were aged between 24 and 35 months (28.4%). Regarding vaccination status, 77% of the cases had received 1 to 4 doses of the Oral Polio (OPV) vaccine. About 62% of the viruses isolated were identified as Cox B, and Echo 3, 6, 7, 11, 12, 14 and 29. Of all NPEVs 37.1% yielded no neutralization pattern. Only 9 (9.67%) cases were followed up; out of which 3 (33.3%) had residual paralysis with one of those with residual paralysis who later died. Conclusion: AFP surveillance and follow up of cases is carried out in Zambia. However, rehabilitation information of the affected children is not followed up, an issue which WHO country office with regards to the transformation agenda could pursue to ensure that the affected children are adequately supported as a contribution to the polio eradication end game. [ABSTRACT FROM AUTHOR]

Acute gastroenteritis is a major cause of childhood morbidity and mortality worldwide. Rotavirus (RV) and Norovirus (NoV) are the leading cause of the disease. Despite the use of improved diagnostic methods a significant proportion of gastroenteritis cases remained undiagnosed. Though nonpolio enteroviruses (NPEVs) have been reported frequently in children with acute gastroenteritis, their etiologic role has not been established. To investigate the epidemiology of NPEVs in gastroenteritis cases which remained negative for leading causative agents, 955 RV and NoV negative stool specimens from children hospitalized for acute gastroenteritis were included in the study. A case control study was conducted which includes stool specimens from 450 children with gastroenteritis and 162 asymptomatic control subjects to determine the association of NPEVs with the disease. NPEV detection and typing was carried out by RT-PCR and sequencing. Presence of RV, NoV, Adenovirus, and Astrovirus was confirmed by ELISA or PCR/RT-PCR. Overall 14% NPEV prevalence was noted. The percentage of children with NPEV infection differed significantly between gastroenteritis and non-gastroenteritis patients (13.7% vs. 4.9%). NPEV was more prevalent among patients with gastroenteritis of undetectable etiology as compared to those detected positive for other viruses (17.9% vs. 7%) ( P < 0.01). Genotyping of NPEV identified predominance of EV-B species (56.5%) followed by EV-C (16.7%), EV-A (13.8%) species and mixed NPEV infections (13%). These data support the association of NPEVs with acute gastroenteritis and highlights the clinical and epidemiological features of NPEV infections in patients with acute gastroenteritis from western India. J. Med. Virol. 87: 154-161, 2015. © 2014 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Human enteroviruses are associated with various clinical syndromes from minor febrile illness to severe, potentially fatal conditions like aseptic meningitis, paralysis, myocarditis, and neonatal enteroviral sepsis. Between June 2000 and August 2008 echovirus (E) type 2, 4, 6, 7, 9, 11, 13, 25, 30, coxsackievirus (CV) -A16, -A19, -B5, and enterovirus 71 (EV71) were reported in Hungary. In this study, 29 previously enterovirus positive samples from 28 patients diagnosed with hand, foot and mouth disease, meningitis and encephalitis, were molecularly typed. The genetic relationships of identified serotypes CV-A16, EV71, and E30 were assessed by direct sequencing of genomic region encoding the capsid protein VP1. The sequences were compared to each other and sequences from other geographical regions possessed in Genbank. The phylogenetic analysis of CV-A16 revealed that the viruses were mostly of Far-Eastern or Asia-Pacific origin. Typing of EV71 showed that one virus from 2000 belonged to genotype C1 and five viruses observed in 2004 and 2005 were identified as genotype C4. The 11 echovirus 30 strains showed homology with those of neighbor European countries. The molecular examination of E30 revealed that three separate lineages circulated in 2000, 2001, and 2004–2006 in Hungary. [ABSTRACT FROM AUTHOR]

Surveillance for acute flaccid paralysis syndrome (AFP) in children under 15 is the backbone of the Global Polio Eradication Initiative. Laboratory examination of stool samples from AFP cases allows the detection of, along with polioviruses, a variety of non-polio enteroviruses (NPEV). The etiological significance of these viruses in the occurrence of AFP cases has been definitively established only for enteroviruses A71 and D68. Enterovirus Coxsackie A2 (CVA2) is most often associated with vesicular pharyngitis and hand, foot and mouth disease. Among 7280 AFP cases registered in Russia over 20 years (2001–2020), CVA2 was isolated only from five cases. However, these included three children aged 3 to 4 years, without overt immune deficiency, immunized with 4–5 doses of poliovirus vaccine in accordance with the National Vaccination Schedule. The disease resulted in persistent residual paralysis. Clinical and laboratory data corresponded to poliomyelitis developing during poliovirus infection. These findings are compatible with CVA2 being the cause of AFP. Molecular analysis of CVA2 from these patients and a number of AFP cases in other countries did not reveal association with a specific phylogenetic group, suggesting that virus genetics is unlikely to explain the pathogenic profile. The overall results highlight the value of AFP surveillance not just for polio control but for studies of uncommon AFP agents. [ABSTRACT FROM AUTHOR]

Abstract: Background: The 66 serotypes of human enteroviruses (EVs) are classified into four species A–D, based on phylogenetic relationships in multiple genome regions. Partial VP1 amplification and sequence analysis are reliable methods for identifying non-polio enterovirus serotypes, especially in negative cell culture specimens from patients with residual paralysis. Objectives: In Iran during the years 2000–2002, there were 29 residual paralysis cases with negative cell (RD, HEp2 and L20B) culture results. Study design: The genomic RNA was extracted from stool specimens from cases of residual paralysis and detected by amplification of the 5′-nontranslated region using RT-PCR with Pan-EV primers. Partial VP1 amplification by semi-nested RT-PCR (snRT-PCR) and sequence analysis were done. Results: Specimens from the 29 culture-negative cases contained echoviruses of six different serotypes. Conclusions: The global eradication of wild polioviruses is near and study of non-polio enteroviruses, which can cause poliomyelitis, is increasingly important to understand their pathogenesis. The VP1 sequences, derived from the snRT-PCR products, allowed rapid molecular analysis of these non-polio strains. [Copyright &y& Elsevier]

BackgroundEnterovirus D68 (EV-D68) has caused major outbreaks of severe respiratory illness worldwide since 2010.AimOur aim was to evaluate EV-D68 circulation in the Netherlands by conducting a serosurvey of EV-D68 neutralising antibodies (nAb) among the Dutch general population.MethodsWe screened 280 sera from children and adults in the Netherlands and used two independent sets of samples collected in the years 2006 and 2007 and in the years 2015 and 2016, time points before and after the first EV-D68 upsurge in 2010. Neutralisation capacity of the sera was tested against the prototype Fermon EV-D68 strain isolated in 1962 and against a recent EV-D68 strain (genotype B3) isolated in France in 2016.ResultsRegardless of the time of serum collection, we found remarkably high overall seropositivity (94.3-98.3%) for nAb against both EV-D68 strains. Geometric mean titres increased in an age-dependent manner.ConclusionsOur data suggest that EV-D68 has been circulating in the Netherlands for decades and that the enterovirus surveillance does not accurately capture the prevalence of this clinically relevant pathogen.