RESUMO
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.
RESUMO
BACKGROUND: Acute flaccid paralysis (AFP) surveillance has been adopted globally as a key strategy for monitoring the progress of the polio eradication initiative. Hereby, to evaluate the completeness of the ascertainment of AFP cases in Italy, a hospital-discharges based search was carried out. METHODS: AFP cases occurring between 2007 and 2016 among children under 15 years of age were searched in the Italian Hospital Discharge Records (HDR) database using specific ICD-9-CM diagnostic codes. AFP cases identified between 2015 and 2016 were then compared with those notified to the National Surveillance System (NSS). RESULTS: Over a 10-year period, 4163 hospital discharges with diagnosis of AFP were reported in Italy. Among these, 956 (23.0%) were acute infective polyneuritis, 1803 (43.3%) myopathy, and 1408 (33.8%) encephalitis, myelitis and encephalomyelitis. During the study period, a decreasing trend was observed for all diagnoses and overall the annual incidence rate (IR) declined from 5.5 to 4.5 per 100,000 children. Comparing NSS with HDR data in 2015-2016, we found a remarkable underreporting, being AFP cases from NSS only 14% of those recorded in HDR. In particular, the acute infective polyneuritis cases reported to NSS accounted for 42.6% of those detected in HDR, while only 0.9% of myopathy cases and 13.1% of encephalitis/myelitis/encephalomyelitis cases have been notified to NSS. The highest AFP IRs per 100,000 children calculated on HDR data were identified in Liguria (17.4), Sicily (5.7), and Veneto (5.1) Regions; regarding the AFP notified to the NSS, 11 out of 21 Regions failed to reach the number of expected cases (based on 1/100,000 rate), and the highest discrepancies were observed in the Northern Regions. Overall, the national AFP rate was equal to 0.6, therefore did not reach the target value. CONCLUSIONS: AFP surveillance data are the final measure of a country's progress towards polio eradication. The historical data obtained by the HDR have been useful to assess the completeness of the notification data and to identify the Regions with a low AFP ascertainment rate in order to improve the national surveillance system.
Assuntos
Paralisia/epidemiologia , Alta do Paciente/estatística & dados numéricos , Poliomielite/epidemiologia , Vigilância da População , Adolescente , Criança , Pré-Escolar , Feminino , Registros Hospitalares , Humanos , Lactente , Itália/epidemiologia , Masculino , Paralisia/virologia , Poliomielite/complicaçõesRESUMO
The WHO Regional Commission for the Certification of Poliomyelitis Eradication has recently indicated Bosnia and Herzegovina (B&H) as a high risk country for transmission, following importation, of wild poliovirus (WPV) or circulating vaccine-derived poliovirus (cVDPV). We analyzed data on Acute Flaccid Paralysis (AFP) surveillance between 2007 to 2016, and the trend of polio immunization coverage in B&H. The majority of AFP cases was recorded in 2016 suggesting an enhancement of the AFP surveillance activities. However, the decline in the immunization coverage, around 74%, and the isolation of two Sabin-like poliovirus type 2 strains, one of them close to a VDPV, require a particular attention in the area. Although B&H has successfully maintained its polio-free status since 2002 several challenges need to be addressed.
Assuntos
Poliomielite/epidemiologia , Poliomielite/virologia , Vacina Antipólio Oral/efeitos adversos , Poliovirus/classificação , Poliovirus/isolamento & purificação , Adolescente , Bósnia e Herzegóvina/epidemiologia , Criança , Pré-Escolar , Monitoramento Epidemiológico , Humanos , Lactente , Recém-NascidoRESUMO
We report the presence of Echovirus 11 (E11) in wastewater in Sicily (Southern Italy), since August 2022. Overall, the 5.4 % of sewage samples (7/130) collected in 2022 were positives for E11 and then the percentage of E11-positive sewage samples reached the value of 27.27(18/66) in the first semester of 2023. Phylogenetic analysis of VP1 sequences showed for most E11-positive samples (16/25: 64 %) close genetic correlation (98.4-99.4 % nucleotide identity) to E11 lineage 1 strains involved in recently reported severe neonatal infections.
Assuntos
Enterovirus , Águas Residuárias , Humanos , Recém-Nascido , Esgotos , Sicília , Filogenia , Enterovirus Humano B/genéticaRESUMO
Innate immune responses to coronavirus infections are highly cell specific. Tissue-resident macrophages, which are infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in patients but are inconsistently infected in vitro, exert critical but conflicting effects by secreting both antiviral type I interferons (IFNs) and tissue-damaging inflammatory cytokines. Steroids, the only class of host-targeting drugs approved for the treatment of coronavirus disease 2019 (COVID-19), indiscriminately suppress both responses, possibly impairing viral clearance. Here, we established in vitro cell culture systems that enabled us to separately investigate the cell-intrinsic and cell-extrinsic proinflammatory and antiviral activities of mouse macrophages infected with the prototypical murine coronavirus MHV-A59. We showed that the nuclear factor κB-dependent inflammatory response to viral infection was selectively inhibited by loss of the lysine demethylase LSD1, which was previously implicated in innate immune responses to cancer, with negligible effects on the antiviral IFN response. LSD1 ablation also enhanced an IFN-independent antiviral response, blocking viral egress through the lysosomal pathway. The macrophage-intrinsic antiviral and anti-inflammatory activity of Lsd1 inhibition was confirmed in vitro and in a humanized mouse model of SARS-CoV-2 infection. These results suggest that LSD1 controls innate immune responses against coronaviruses at multiple levels and provide a mechanistic rationale for potentially repurposing LSD1 inhibitors for COVID-19 treatment.
Assuntos
COVID-19 , Lisina , Animais , Humanos , Camundongos , Antivirais/farmacologia , Tratamento Farmacológico da COVID-19 , Citocinas/metabolismo , SARS-CoV-2/metabolismoRESUMO
BACKGROUND: Acute Flaccid Paralyses Surveillance (AFPS) monitors the emergence of polioviruses and can track Non-Polio Enteroviruses (NPEVs). We report AFPS activity in the Lombardy region (Northern Italy) from 2016 to 2018. METHODS: Fecal and respiratory samples were collected from children <15 years who met the WHO definition of an AFP case, analyzed by virus isolation in cell cultures (RD/L20B) and by a one-step real-time RT-PCR assay specific for the 5'-noncoding-region of NPEV. NPEV-positive specimens were further analyzed by sequencing a fragment of the VP1 gene. RESULTS: 36 AFP cases (89 stool and 32 respiratory samples) were reported with an incidence of 1.1/100'000, 0.9/100'000, 0.6/100'000 children <15 years in 2016, 2017, 2018, respectively. Poliovirus was not identified, whereas NPEVs were detected in 19.4% (7/36) of AFP cases. The presence of one Echovirus-25 (2016), two EV- and D68 (2016 and 2018), one EV-A71 (2016), and one Echovirus-30 (2016) sharing high nucleotide identity with NPEVs detected in Europe was identified. CONCLUSION: The absence of polio was confirmed. The unpredicted detection of emerging EV-D68, EV-A71, and E-30 sharing high sequence nucleotide similarity with viruses involved in the latest outbreaks, provided valuable and up-to-date information, emphasizing the importance of monitoring NPEVs through AFPS.
Assuntos
Viroses do Sistema Nervoso Central/epidemiologia , Viroses do Sistema Nervoso Central/virologia , Enterovirus/isolamento & purificação , Enterovirus/fisiologia , Monitoramento Epidemiológico , Mielite/epidemiologia , Mielite/virologia , Doenças Neuromusculares/epidemiologia , Doenças Neuromusculares/virologia , Criança , Pré-Escolar , Fezes/virologia , Feminino , Humanos , Incidência , Lactente , Itália/epidemiologia , MasculinoRESUMO
Polyphenols have been widely studied for their antiviral effect against respiratory virus infections. Among these, resveratrol (RV) has been demonstrated to inhibit influenza virus replication and more recently, it has been tested together with pterostilbene against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In the present work, we evaluated the antiviral activity of polydatin, an RV precursor, and a mixture of polyphenols and other micronutrients, named A5+, against influenza virus and SARS-CoV-2 infections. To this end, we infected Vero E6 cells and analyzed the replication of both respiratory viruses in terms of viral proteins synthesis and viral titration. We demonstrated that A5+ showed a higher efficacy in inhibiting both influenza virus and SARS-CoV-2 infections compared to polydatin treatment alone. Indeed, post infection treatment significantly decreased viral proteins expression and viral release, probably by interfering with any step of virus replicative cycle. Intriguingly, A5+ treatment strongly reduced IL-6 cytokine production in influenza virus-infected cells, suggesting its potential anti-inflammatory properties during the infection. Overall, these results demonstrate the synergic and innovative antiviral efficacy of A5+ mixture, although further studies are needed to clarify the mechanisms underlying its inhibitory effect.
RESUMO
BACKGROUND: Enterovirus infections can cause a variety of illnesses, ranging from asymptomatic infections to severe illness and death. AIM: To support polio eradication activities, in February 2019, the WHO Regional Reference Laboratory for polio in Italy, at the National Institute of Public Health (Istituto Superiore di Sanità), promoted an investigation on non-polio enterovirus laboratory capacity, with the support of the Italian Ministry of Health. The aim was to collect data on the assays used routinely for diagnostic purposes and to characterize enterovirus outbreaks strains by sequence analysis of the Viral Protein 1 region. METHODS: A questionnaire was administered to public health laboratories through all Italian Regions for 2018 and subsequently, an electronic form for lab-confirmed enterovirus infection reported from February 2019 to January 2020, including patients clinical characteristics, and laboratory data was distributed through 25 laboratories participating the survey. RESULTS: Overall, a homogenous laboratory capacity for enterovirus infection diagnosis was found and 21,000 diagnostic tests were retrospectively reported in 2018. Then, in 2019, two outbreaks of Echovirus 30 were identified and confirmed by molecular analyses. CONCLUSION: These results underline the need monitor the circulation of non-polio enterovirus to ascertain the real burden of the disease in the country.
Assuntos
Infecções por Enterovirus , Surtos de Doenças , Enterovirus Humano B/genética , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/epidemiologia , Humanos , Itália/epidemiologia , Estudos Retrospectivos , Inquéritos e QuestionáriosRESUMO
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.
RESUMO
Background: Neonatal acute liver failure (ALF) is a severe condition with a mortality rate up to 70%. Human enterovirus (HEV) infections are associated with serious diseases in newborns, including myocarditis, meningoencephalitis and, more rarely, ALF with a fulminant course.Methods: Cases of neonatal-onset ALF were identified using the institutional clinical database. The history and clinical data of infants with HEV infection were collected by medical record revision. Viral testing by nested real- time PCR (nRT-PCR) was performed by the Bambino Gesù Children's Hospital Clinical Laboratory and by National Institute of Public Health in Rome.Results: Among ten infants referred to our Institution with neonatal-onset ALF in the 2004-2018 period, we identified five cases due to HEV. In three of these, the mother reported an episode of mild fever and diarrhea during the last trimester of gestation, suggesting fetal-maternal transmission. All were late preterm infants (32-36 weeks). Two infants died as a result of ALF; the other three survived with full normalization of liver function. In four, the causing agents were coxsackie B serotypes 3 (n = 1), 4 (n = 1) and 5 (n = 2), in the fifth case we identified echovirus serotype 11.Conclusions: Human enterovirus (HEV) are a rare but relevant cause of ALF in neonates. HEV testing should be systematically performed in cases of neonatal ALF for diagnostic and management purposes.
Assuntos
Infecções por Enterovirus , Enterovirus , Falência Hepática Aguda , Criança , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/epidemiologia , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Unidades de Terapia Intensiva Neonatal , Falência Hepática Aguda/diagnóstico , Falência Hepática Aguda/epidemiologia , Falência Hepática Aguda/etiologiaRESUMO
Oral poliovirus vaccine (OPV) can cause, in extremely rare cases vaccine-associated paralytic poliomyelitis in recipients, or contacts of vaccinees. Three cases of vaccine-associated paralytic poliomyelitis (two contacts and one recipient) occurred in the Bourgas region of Bulgaria in the spring of 2006. The first two cases, notified as acute flaccid paralysis, were 55 days old unvaccinated twin brothers, having been in contact with vaccinees. The third case concerned a 4-month-old infant who had received the first OPV dose 37 days prior to the onset of illness. Complete clinical, epidemiological, virological, serological and molecular investigations of the children with paralysis and their contacts were undertaken. In all the three cases type 3 polioviruses were isolated from fecal samples and characterized as Sabin-like poliovirus strains. Type 3 polioviruses isolated from the twin brothers demonstrated by sequence analysis U-to-C back mutation at nt 472 of the 5' UTR, known to correlate with neurovirulence, and mutation in the VP1 region. Type 3 poliovirus isolated from the third child demonstrated in the 3D sequenced region a recombination with Sabin type 1 poliovirus. In the latter region, three silent mutations and one, resulting in amino acid substitution, were also observed. The clinical, epidemiological and virological data and the neurological sequelae observed 60 days following the onset of paralysis, confirmed the diagnosis of vaccine-associated paralytic poliomyelitis in all the three patients.
Assuntos
Poliomielite/virologia , Vacinas contra Poliovirus/efeitos adversos , Poliovirus/isolamento & purificação , Regiões 5' não Traduzidas/genética , Adulto , Bulgária , Criança , Pré-Escolar , Fezes/virologia , Humanos , Lactente , Recém-Nascido , Masculino , Mutação Puntual , Poliomielite/patologia , Poliovirus/genética , Recombinação Genética , Análise de Sequência de DNARESUMO
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.
Assuntos
Enterovirus Humano B/genética , Esgotos/virologia , Enterovirus Humano B/classificação , Enterovirus Humano B/isolamento & purificação , Monitoramento Ambiental , Itália , Filogenia , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
Human enteroviruses (HEVs) occur in high concentrations in wastewater and can contaminate receiving environmental waters, constituting a major cause of acute waterborne disease worldwide. In this study, we investigated the relative abundance, occurrence, and seasonal distribution of polio and other enteroviruses at three wastewater treatment plants (WWTPs) in Naples, Southern Italy, from January 2010 to December 2014. Influent and effluent samples from the three WWTPs were collected monthly. One hundred and sixty-one of the 731 wastewater samples collected (22.0%) before and after water treatment were CPE positive on RD cells; while no samples were positive on L20B cells from any WWTPs. Among the 140 non-polio enterovirus isolated from inlet sewage, 69.3% were Coxsackieviruses type B and 30.7% were Echoviruses. Among these, CVB3 and CVB5 were most prevalent, followed by CVB4 and Echo6. The twenty-one samples tested after treatment contained 6 CVB4, 5 CVB3, 3 Echo11, and 2 Echo6; while other serotypes were isolated less frequently. Data on viral detection in treated effluents of WWTPs confirmed the potential environmental contamination by HEVs and could be useful to establish standards for policies on wastewater management.
Assuntos
Infecções por Enterovirus/virologia , Enterovirus/isolamento & purificação , Águas Residuárias/virologia , Purificação da Água/métodos , Humanos , Itália , Poliomielite/virologia , Poliovirus/isolamento & purificação , Esgotos/virologia , Inquéritos e QuestionáriosRESUMO
Within the initiatives for poliomyelitis eradication by WHO, Italy activated an environmental surveillance (ES) in 2005. ES complements clinical Acute Flaccid Paralysis (AFP) surveillance for possible polio cases, detects poliovirus circulation in environmental sewage, and is used to monitor transmission in communities. In addition to polioviruses, the analyses comprised: (i) the monitoring of the presence of non-polio enteroviruses in sewage samples and (ii) the temporal and geographical distribution of the detected viruses. From 2009 to 2015, 2880 sewage samples were collected from eight cities participating in the surveillance. Overall, 1479 samples resulted positive for enteroviruses. No wild-type polioviruses were found, although four Sabin-like polioviruses were detected. The low degree of mutation found in the genomes of these four isolates suggests that these viruses have had a limited circulation in the population. All non-polio enteroviruses belonged to species B and the most frequent serotype was CV-B5, followed by CV-B4, E-11, E-6, E-7, CV-B3, and CV-B2. Variations in the frequency of different serotypes were also observed in different seasons and/or Italian areas. Environmental surveillance in Italy, as part of the 'WHO global polio eradication program', is a powerful tool to augment the polio surveillance and to investigate the silent circulation or the re-emergence of enteroviruses in the population.
Assuntos
Infecções por Enterovirus/virologia , Enterovirus/imunologia , Poliomielite/virologia , Poliovirus/imunologia , Esgotos/virologia , Cidades , Enterovirus/classificação , Enterovirus/isolamento & purificação , Infecções por Enterovirus/epidemiologia , Monitoramento Ambiental , Humanos , Itália/epidemiologia , Limite de Detecção , Poliomielite/epidemiologia , Poliovirus/classificação , Poliovirus/isolamento & purificaçãoRESUMO
We conducted a study to determine the main traits of pertussis among unimmunized infants less than 6 months of age. From August 2012 to March 2015, 141 nasopharyngeal aspirates (NPAs) were collected from infants with respiratory symptoms attending 2 major hospitals in Rome. Clinical data were recorded and analyzed. Lab-confirmation was performed by culture and realtime PCR. B. pertussis virulence-associated genes (ptxP, ptxA and prn), together with multilocus variable-number tandem repeat analysis (MLVA), were also investigated by the sequence-based analysis on the DNAs extracted from positive samples. Antibiotic susceptibility with Etest was defined on 18 viable B. pertussis isolates. Samples from 73 infants resulted positives for B. pertussis. The median age of the patients was 45 d (range 7-165); 21 infants were treated with macrolides before hospital admission. Cough was reported for a median of 10 d before admission and 18 d after hospital discharge among infected infants, 84% of whom showed paroxysmal cough. No resistance to macrolides was detected. Molecular analysis identified MT27 as the predominant MLVA profile, combined with ptxP3-ptxA1-prn2 associated virulence genes. Although our data may not be generalized to the whole country, they provide evidence of disease severity among infants not vaccinated against pertussis. Moreover, genetically related B. pertussis strains, comprising allelic variants of virulence associated genes, were identified.
Assuntos
Bordetella pertussis/genética , Coqueluche/epidemiologia , Coqueluche/microbiologia , Antibacterianos/farmacologia , Bordetella pertussis/efeitos dos fármacos , Bordetella pertussis/isolamento & purificação , Bordetella pertussis/patogenicidade , DNA Bacteriano/genética , Feminino , Humanos , Lactente , Recém-Nascido , Itália/epidemiologia , Macrolídeos/farmacologia , Masculino , Tipagem de Sequências Multilocus , Nasofaringe/microbiologia , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Virulência/genéticaRESUMO
The global eradication of polio is close to achieving success. However, transmission of wild poliovirus persists in countries where the disease is endemic, and outbreaks may also occur in previously polio-free countries where population immunity is not maintained. To achieve polio eradication, several key actions are required: (1) to detect any poliovirus transmission, (2) to strengthen immunization systems and withdraw all oral polio vaccines from use, (3) to contain polioviruses and certify interruption of transmission, and (4) to sustain ongoing public health programs. In this context, residual hurdles to global polio eradication have been identified, from unstable political situation in affected countries to population movements from and to endemic areas, and opposition to immunization strategies. Global efforts are needed in order to promote routine immunization campaigns and large-scale vaccination rounds, which may provide direct protection of individuals and minimize virus circulation.
Assuntos
Poliomielite/prevenção & controle , Doenças Endêmicas/prevenção & controle , Saúde Global , Humanos , Poliomielite/epidemiologia , Vacinas contra PoliovirusRESUMO
We report pertussis cases in 4 infants less than 6 months admitted with symptoms compatible with pertussis to the intensive care unit of the Università Cattolica del Sacro Cuore in Rome, April 2014. Realtime PCR confirmed pertussis diagnosis for the 4 infants, 2 of them were cousins, and for the household contacts of 1 of them. Analysis of pertussis toxin, its promoter and pertactin was also performed. First of all, this report emphasizes the need to investigate household contact of infants with pertussis; secondly, to evaluate the selective vaccination of household members of newborns as an effective program to reduce pertussis in infants.
Assuntos
Características da Família , Saúde da Família , Coqueluche/epidemiologia , Proteínas da Membrana Bacteriana Externa/genética , Bordetella pertussis/genética , Bordetella pertussis/isolamento & purificação , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Toxina Pertussis/genética , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Cidade de Roma/epidemiologia , Fatores de Virulência de Bordetella/genéticaRESUMO
A new rapid method for identification and determination of the titer of polioviruses and other enteroviruses in cell monolayers grown in microtiter plates is described. The method is based on immunoperoxidase staining of infected cells with commercial monoclonal antibodies (MAb) and biotin-labeled secondary antibody. The presence of poliovirus or other enteroviruses was established as the appearance of at least one focus of cells with stained cytoplasm 6 h post-infection. Viral titers determined by this method were expressed as focus forming unit (FFU) per ml which was found to correspond approximately to 10 TCID(50)/ml. The suitability of this technique to determine poliovirus antibody titers in human sera was also tested comparing the immunocytochemical neutralization assay with a conventional neutralization in microtiter plates. The test was standardized using reference human sera in order to produce antibody titers expressed in international units (IU). In addition to high reproducibility, the new neutralization test appears to be sensitive, specific and rapid, and might thus represent a useful tool for the diagnosis of polio and other enterovirus infections.
Assuntos
Anticorpos Antivirais/sangue , Técnicas Imunoenzimáticas/métodos , Testes de Neutralização/métodos , Poliomielite/virologia , Poliovirus/imunologia , Poliovirus/isolamento & purificação , Anticorpos Monoclonais/imunologia , Biotinilação , Linhagem Celular , Humanos , Poliomielite/diagnóstico , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sorotipagem , Fatores de TempoRESUMO
This study evaluated the presence and seasonal distribution of polio and other enteroviruses in four wastewater treatment plants in three cities in Italy, using different treatment systems. Detection of enteroviruses was carried out by virus isolation in cell cultures after concentration of water samples collected at both inlet and outlet of the treatment plants, following the methods described in the WHO guidelines. Viral serotypes isolated before and after water treatment were compared. Forty-eight non-polio enteroviruses were isolated from 312 samples collected at the inlet of the four wastewater treatment plants, 35 of which were Coxsackievirus type B (72.9 %) and 13 Echovirus (27.1 %). After treatment, 2 CVB3, 1 CVB5, and 1 Echo 6 were isolated. CVB3 and Echo 6 serotypes were also detected in samples collected at the inlet of the TP, in the same month and year. The high rate of detection of infectious enteroviruses in inlet sewage samples (30.1 %) indicates wide diffusion of these viruses in the populations linked to the collectors. The incomplete removal of infectious viruses following sewage treatment highlights possible risks for public health relate to treated waters discharge into the environment.
Assuntos
Enterovirus/isolamento & purificação , Águas Residuárias/virologia , Purificação da Água/instrumentação , Enterovirus/classificação , Enterovirus/genética , Itália , Filogenia , Esgotos/virologiaRESUMO
A panel of murine IgG monoclonal antibodies (MAbs) was produced against coxsackievirus A9 (CAV9). Fifty-nine MAbs reactive in ELISA with purified CAV9 were identified. Eighteen of them could efficiently inhibit infection by CAV9 but not coxsackieviruses B. Neutralization-resistant CAV9 variants to four different MAbs were isolated and tested for resistance to neutralization by other MAbs of the panel. Three groups of reactivity including 10, 7, and 1 MAbs were thus identified. Sequencing of neutralization-escape virus mutants showed that neutralization by one MAb group was affected by change of VP3 amino acids 62 or 69. For the second group of reactivity, mutations included amino acids 154 or 165 of VP2. The only MAb of the third group selected for a change at residue 70 of VP2. Protection studies in a newborn mouse model of myositis suggested that either epitopes in VP2 or in VP3 mediate protection from CAV9 infection in vivo.