P. Mancini, D. Brandtner, C. Veneri, G. Bonanno Ferraro, M. Iaconelli, S. Puzelli, M. Facchini, G. Di Mario, P. Stefanelli, L. Lucentini, A. Muratore, The SARI network, E. Suffredini, G. La Rosa
{"title":"Evaluation of Trends in Influenza A and B Viruses in Wastewater and Human Surveillance Data: Insights from the 2022–2023 Season in Italy","authors":"P. Mancini, D. Brandtner, C. Veneri, G. Bonanno Ferraro, M. Iaconelli, S. Puzelli, M. Facchini, G. Di Mario, P. Stefanelli, L. Lucentini, A. Muratore, The SARI network, E. Suffredini, G. La Rosa","doi":"10.1007/s12560-024-09622-2","DOIUrl":"10.1007/s12560-024-09622-2","url":null,"abstract":"<div><p>Wastewater-based epidemiology (WBE) is a recognized, dynamic approach to monitoring the transmission of pathogens in communities through urban wastewater. This study aimed to detect and quantify influenza A and B viruses in Italian wastewater during the 2022–2023 season (October 2022 to April 2023). A total of 298 wastewater samples were collected from 67 wastewater treatment plants (WTPs) across the country. These samples were analyzed for influenza A and B viruses (IAV, IBV) using primers originally developed by the Centers for Disease Control and Prevention (CDC) for real-time PCR and adapted for digital PCR. The overall detection rates of IAV and IBV across the entire study period were 19.1% and 16.8%, respectively. The prevalence of IAV in wastewater showed a gradual increase from October to December 2022, peaking at 61% in December. In contrast, IBV peaked at 36% in February 2023. This temporal discrepancy in peak concentrations suggests different seasonal patterns for the two influenza types. These trends mirrored human surveillance data, which showed influenza A cases peaking at 46% in late December and declining to around 2% by April 2023, and influenza B cases starting to increase significantly in January 2023 and peaking at about 14% in March. IAV concentrations ranged from 9.80 × 10<sup>2</sup> to 1.94 × 10<sup>5</sup> g.c./L, while IBV concentrations ranged from 1.07 × 10<sup>3</sup> to 1.43 × 10<sup>4</sup> g.c./L. Overall, the environmental data were consistent with the human surveillance trends observed during the study period in the country. These results demonstrate the value of WBE in tracking epidemiological patterns and highlight its potential as a complementary tool to infectious diseases surveillance systems.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09622-2.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142789375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kata Farkas, Rachel C. Williams, Luke S. Hillary, Alvaro Garcia-Delgado, Eleanor Jameson, Jessica L. Kevill, Matthew J. Wade, Jasmine M. S. Grimsley, Davey L. Jones
{"title":"Harnessing the Power of Next-Generation Sequencing in Wastewater-Based Epidemiology and Global Disease Surveillance","authors":"Kata Farkas, Rachel C. Williams, Luke S. Hillary, Alvaro Garcia-Delgado, Eleanor Jameson, Jessica L. Kevill, Matthew J. Wade, Jasmine M. S. Grimsley, Davey L. Jones","doi":"10.1007/s12560-024-09616-0","DOIUrl":"10.1007/s12560-024-09616-0","url":null,"abstract":"<div><p>Wastewater-based epidemiology (WBE) has emerged as a valuable surveillance tool for SARS-CoV-2 and other pathogens globally, providing insights into community-level infections, including asymptomatic and pre-symptomatic cases. While most WBE programmes focus on quantitative pathogen assessment, next-generation sequencing (NGS) approaches have enabled more detailed analyses, including variant and recombinant genotype identification for viruses like SARS-CoV-2 and poliovirus. Despite recent NGS advancements allowing for the detection of known and novel viruses in wastewater, many of these tools remain underutilised in routine WBE. This short review critically evaluates the applicability of common NGS tools in routine WBE programmes, assessing their capability for identifying emerging threats with epidemic or pandemic potential. Here, we provide evidence-based recommendations for integrating NGS techniques into WBE and the use of results for informed decision-making within a One Health framework, aiming to enhance global infectious disease surveillance and pandemic preparedness.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09616-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142753935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Priscilla Gomes da Silva, José Gonçalves, Elisa Rodriguéz, Pedro A. García-Encina, Maria São José Nascimento, Sofia I. V. Sousa, João R. Mesquita
{"title":"SARS-CoV-2 RNA Presence in Outdoor Air of Public Spaces in Valladolid During Winter, 2021","authors":"Priscilla Gomes da Silva, José Gonçalves, Elisa Rodriguéz, Pedro A. García-Encina, Maria São José Nascimento, Sofia I. V. Sousa, João R. Mesquita","doi":"10.1007/s12560-024-09615-1","DOIUrl":"10.1007/s12560-024-09615-1","url":null,"abstract":"<div><p>As SARS-CoV-2 continues to evolve and herd immunity establishes, an increasing number of asymptomatic infections have been reported, increasing the risk of airborne spread of the virus. Most of the studies regarding SARS-CoV-2 RNA presence in air refer to indoor environments, with few studies having reported SARS-CoV-2 RNA in outdoor air. The aim of this study was to assess the presence of SARS-CoV-2 RNA at two different settings, crowded outdoor versus empty outdoor environments in Valladolid, Spain, during winter 2021. Using a Coriolis® air sampler, samples were taken from nine different locations within the city center. RNA extraction and a one-step RT-qPCR were carried out. Six out of the 20 air samples were found to be positive, and they were all obtained from crowded outdoor environments. These results highlight that although in less quantity, SARS-CoV-2 RNA is still present in outdoor air, especially at moments of relaxed mitigation efforts and depending on the number of people present.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09615-1.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142753968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sibiri Sylvain Rouamba, François Tapsoba, Boukaré Kaboré, Adama Patrice Soubeiga, Moumouni Bandé, Elie Kabré, Aly Savadogo
{"title":"Impact of Wastewater Use for Irrigation and Contamination of Lettuce by Enteric Viruses: Case of Ouagadougou Market Gardening Sites, Burkina Faso","authors":"Sibiri Sylvain Rouamba, François Tapsoba, Boukaré Kaboré, Adama Patrice Soubeiga, Moumouni Bandé, Elie Kabré, Aly Savadogo","doi":"10.1007/s12560-024-09621-3","DOIUrl":"10.1007/s12560-024-09621-3","url":null,"abstract":"<div><p>Raw vegetables irrigated with polluted water that may contain enteric viruses can be associated with foodborne viral disease outbreaks. The objective of this study is to investigate the possible transmission of enteric viruses from irrigation water to lettuce. Therefore, we performed a commercial multiplex real-time PCR assay to monitor the occurrence of enteric viruses in irrigation water samples and in raw vegetables that were cultivated at market gardening sites in Ouagadougou, Burkina Faso. Samples were collected from six market gardening sites located in Ouagadougou. RT-PCR was performed to detect norovirus GI, norovirus GII, rotavirus, enteric adenoviruses F (Serotype 40/41), astrovirus and sapovirus (Genogroups G1, 2, 4, 5). From the 10 irrigation water samples and the 80 lettuce samples, three (30%) and twenty-two (27.5%) were positive for enteric viruses, respectively. Norovirus GII, astrovirus and enteric adenoviruses F (Serotype 40/41) were the most frequently detected viruses in lettuce and irrigation water samples. Our results indicate that raw vegetables may be contaminated with a broad range of enteric viruses, which may originate from virus-contaminated irrigation water, and these vegetables may act as a potential vector of food-borne viral transmission.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-11-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guadalupe Di Cola, Anabella C. Fantilli, Gonzalo Rodríguez-Lombardi, Kevin A. Rucci, Gonzalo Castro, Santiago Mirazo, Silvia Viviana Nates, María Belén Pisano, Viviana E. Ré
{"title":"Assessment of Hepatitis E Virus RNA Detection in Meat Samples: Optimization of Pre-analytical Conditions","authors":"Guadalupe Di Cola, Anabella C. Fantilli, Gonzalo Rodríguez-Lombardi, Kevin A. Rucci, Gonzalo Castro, Santiago Mirazo, Silvia Viviana Nates, María Belén Pisano, Viviana E. Ré","doi":"10.1007/s12560-024-09617-z","DOIUrl":"10.1007/s12560-024-09617-z","url":null,"abstract":"<div><p>Hepatitis E virus (HEV) is primarily transmitted via the fecal–oral route and is considered an anthropozoonosis. Genotypes with zoonotic potential (mainly HEV-3 and HEV-4) can be transmitted through the consumption of raw or undercooked pork, wild boar, deer meat, or processed products. This study aims to explore methodologies for processing meat samples to establish a protocol for HEV detection in meat. The analysis of pre-analytical conditions involved comparing homogenization with PBS versus TRIzol, comparing tissue disruption methods (ultra-turrax versus mortar and pestle), and assessing nucleic acid extraction techniques (spin columns and magnetic beads) across three types of artificially contaminated meat matrices: pork, salmon (fish-meat), and salami. Each test included a process control virus (PP7) and an HEV transcript. Molecular detection was performed via RT-qPCR. Results indicated that TRIzol provided better recovery rates for homogenization, while spin columns were the most effective option for RNA extraction. Both the ultra-turrax homogenizer and the mortar-pestle methods were effective for pork and fish-meat homogenization, while the use of the UT yielded superior results for salami. HEV recovery rates were 36.7%, 26.3%, and 34.1% for salami, salmon, and pork meat, respectively. In conclusion, we reached a simple and reliable protocol for the detection of RNA-HEV from three meat matrices. This method, which includes homogenization with TRIzol, mechanical tissue disruption, and RNA extraction using spin columns followed by real-time PCR, can be applied in future studies to evaluate HEV prevalence in food sources and contribute to the discussion about HEV detection methodologies.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142691841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Monitoring Influenza A (H1N1, H3N2), RSV, and SARS-CoV-2 Using Wastewater-Based Epidemiology: A 2-Year Longitudinal Study in an Indian Megacity Covering Omicron and Post-Omicron Phases","authors":"Rinka Pramanik, Kiran Nannaware, Vinita Malik, Priyanki Shah, Poornima Sangewar, Niharika Gogate, L. S. Shashidhara, Radhika Boargaonkar, Dhawal Patil, Saurabh Kale, Asim Bhalerao, Nidhi Jain, Sanjay Kamble, Syed Dastager, Mahesh Dharne","doi":"10.1007/s12560-024-09618-y","DOIUrl":"10.1007/s12560-024-09618-y","url":null,"abstract":"<div><p>The bourgeoning field of wastewater-based epidemiology (WBE) for the surveillance of several respiratory viruses which includes Influenza A, H1N1pdm09, H3N2, respiratory syncytial viruses (RSV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of interest for public health concerns. However, there are few long-term monitoring studies globally. In this study, respiratory viruses were detected and quantified from 11 sewer sheds by utilizing reverse transcription-quantitative polymerase chain reaction analysis in Pune city, India, from Jan 2022 to Dec 2023. The RNA fragments of respiratory viruses were detected in sewage samples before clinical cases were reported, underscoring the potential of WBE for early detection and monitoring within the population. The Spearman correlation of wastewater viral copies was positively and significantly correlated with the clinically positive case of H1N1pdm09 (<i>ρ</i> = 0.55, <i>p</i> = 1.4 × 10<sup>–9</sup>), H3N2 (<i>ρ</i> = 0.25, <i>p</i> = 9.9 × 10<sup>–3</sup>), and SARS-CoV-2 (<i>ρ</i> = 0.43, <i>p</i> = 4.1 × 10<sup>–6</sup>). The impact of public health interventions on the circulation of infectious respiratory diseases showed a significant difference in the viral load during the period when many preventing measures were carried out against the COVID-19 pandemic (restriction phase), compared to the period when no such preventive measures are followed (no-restriction phase) for Influenza A, H1N1pdm09, H3N2, and RSV with <i>p</i>-value < 0.05, which indicates the influence of health policy implementation in controlling disease spread. The present study provides an effective approach to detecting multiple respiratory viruses from wastewater and provides insights into the epidemiology of respiratory illnesses. The WBE aids in providing information on the spread of pathogens (viruses) in the community, offering a proactive strategy for public health management, allowing for timely interventions and implementing targeted measures to mitigate the spread of these viruses under one health approach.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"17 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Ultraviolet (UV-C) Light Systems for the Inactivation of Feline Calicivirus and Tulane Virus in Model Fluid Foods","authors":"E. Corson, B. Pendyala, A. Patras, D. H. D’Souza","doi":"10.1007/s12560-024-09614-2","DOIUrl":"10.1007/s12560-024-09614-2","url":null,"abstract":"<div><p>Conventional UV-C (254 nm) inactivation technologies have limitations and potential operator-safety risk. To overcome these disadvantages, novel UV-C light-emitting diodes (LED) are developed and investigated for their performance. This study aimed to determine the inactivation of human norovirus (HuNoV) surrogates, Tulane virus (TV), and feline calicivirus (FCV-F9), by UV-C (254 nm) in comparison to UV-C LED (279 nm) in phosphate-buffered saline (PBS) and coconut water (CW). Five-hundred microliters of FCV-F9 (~ 5 log plaque forming units (PFU)/mL) or TV (~ 6 log PFU/mL) were added to 4.5 mL PBS or CW in continuously stirred glass beakers and exposed to 254 nm UV-C for 0 up to 15 min (maximum dosage of 33.89 mJ/cm<sup>2</sup>) or 279 nm UV-C LED for 0 up to 2.5 min (maximum dosage of 7.03 mJ/cm<sup>2</sup>). Recovered viruses were assayed in duplicate from each treatment replicated thrice. Mixed model analysis of variance was used for data analysis. Significantly lower D<sub>10</sub> values were obtained in PBS and CW (<i>p</i> ≤ 0.05) for both tested viruses using UV-C LED (279 nm) where FCV-F9 showed D<sub>10</sub> values of 7.08 ± 1.75 mJ/cm<sup>2</sup> and 3.75 ± 0.11 mJ/cm<sup>2</sup>, while using UV-C (254 nm) showed D<sub>10</sub> values of 13.81 ± 0.40 mJ/cm<sup>2</sup> and 6.43 ± 0.44 mJ/cm<sup>2</sup> in PBS and CW, respectively. Similarly, lower D<sub>10</sub> values were obtained for TV of 3.91 ± 1.03 mJ/cm<sup>2</sup> and 4.26 ± 1.02 mJ/cm<sup>2</sup> with 279 nm UV-C LED and were 18.76 ± 3.16 mJ/cm<sup>2</sup> and 10.21 ± 1.48 mJ/cm<sup>2</sup> with 254 nm UV-C in PBS and CW, respectively. Viral resistance to these treatments was fluid-matrix dependent. These findings indicate that use of 279 nm UV-C LED is more effective in inactivating HuNoV surrogates than conventional 254 nm UV-C in the tested fluids.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"16 4","pages":"506 - 515"},"PeriodicalIF":4.1,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142387174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Allyson N. Hamilton, Flor Maes, Génesis Yosbeth Chávez Reyes, Giselle Almeida, Dan Li, Mieke Uyttendaele, Kristen E. Gibson
{"title":"Machine Learning and Imputation to Characterize Human Norovirus Genotype Susceptibility to Sodium Hypochlorite","authors":"Allyson N. Hamilton, Flor Maes, Génesis Yosbeth Chávez Reyes, Giselle Almeida, Dan Li, Mieke Uyttendaele, Kristen E. Gibson","doi":"10.1007/s12560-024-09613-3","DOIUrl":"10.1007/s12560-024-09613-3","url":null,"abstract":"<div><p>Human norovirus (HuNoV) is the leading cause of foodborne illness in the developed world and a major contributor to gastroenteritis globally. Its low infectious dose and environmental persistence necessitate effective disinfection protocols. Sodium hypochlorite (NaOCl) bleach is a widely used disinfectant for controlling HuNoV transmission via contaminated fomites. This study aimed to evaluate the susceptibility of HuNoV genotypes (<i>n</i> = 11) from genogroups I, II, and IV to NaOCl in suspension. HuNoV was incubated for 1 and 5 min in diethyl pyrocarbonate (DEPC) treated water containing 50 ppm, 100 ppm, or 150 ppm NaOCl, buffered to maintain a pH between 7.0 and 7.5. Neutralization was achieved by a tenfold dilution into 100% fetal bovine serum. RNase pre-treatment followed by RT-qPCR was used to distinguish between infectious and non-infectious HuNoV. Statistical methods, including imputation, machine learning, and generalized linear models, were applied to process and analyze the data. Results showed that NaOCl reduced viral loads across all genotypes, though efficacy varied. Genotypes GI.1, GII.4 New Orleans, and GII.4 Sydney were the least susceptible, while GII.6 and GII.13 were the most susceptible. All NaOCl concentrations above 0 ppm were statistically indistinguishable, and exposure duration did not significantly affect HuNoV reduction, suggesting rapid inactivation at effective concentrations. For instance, some genotypes were completely inactivated within 1 min, rendering extended exposure unnecessary, while other genotypes maintained the initial concentration at both 1 and 5 min, indicating a need for longer contact times. These findings underscore the critical role of HuNoV genotype selection in testing disinfection protocols and optimizing NaOCl concentrations. Understanding HuNoV susceptibility to NaOCl bleach informs better disinfection strategies, aiding public health and food safety authorities in reducing HuNoV transmission and outbreaks.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"16 4","pages":"492 - 505"},"PeriodicalIF":4.1,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09613-3.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. Scott, D. Ryder, M. Buckley, R. Hill, S. Treagus, T. Stapleton, D. I. Walker, J. Lowther, F. M. Batista
{"title":"Long Amplicon Nanopore Sequencing for Dual-Typing RdRp and VP1 Genes of Norovirus Genogroups I and II in Wastewater","authors":"G. Scott, D. Ryder, M. Buckley, R. Hill, S. Treagus, T. Stapleton, D. I. Walker, J. Lowther, F. M. Batista","doi":"10.1007/s12560-024-09611-5","DOIUrl":"10.1007/s12560-024-09611-5","url":null,"abstract":"<div><p>Noroviruses (NoVs) are the leading cause of non-bacterial gastroenteritis with societal costs of US$60.3 billion per annum. Development of a long amplicon nanopore-based method for dual-typing the <i>RNA-dependent RNA polymerase</i> (<i>RdRp</i>) and major structural protein (<i>VP1</i>) regions from a single RNA fragment could improve existing norovirus typing methods. Application to wastewater-based epidemiology (WBE) and environmental testing could enable the discovery of novel types and improve outbreak tracking and source apportionment. Here, we have developed such a method with a consensus-based bioinformatics pipeline and optimised reverse transcription (RT) and PCR procedures. Inhibitor removal and LunaScript® RT gave robust amplification of the ≈ 1000 bp <i>RdRP</i> + <i>VP1</i> amplicon for both the GI and GII PCR assays. Platinum™ Taq polymerase showed good sensitivity and reduced levels non-specific amplification (NSA) when compared to other polymerases. Optimised PCR annealing temperatures significantly reduced NSA (51.3 and 42.4% for GI and GII), increased yield (86.5% for GII) and increased taxa richness (57.7%) for GII. Analysis of three NoV positive faecal samples showed 100% nucleotide similarity with Sanger sequencing. Eight GI genotypes, 11 polymerase types (p-types) and 13 combinations were detected in wastewater along with 4 GII genotypes, 4 p-types and 8 combinations; highlighting the diversity of norovirus taxa present in wastewater in England. The most common genotypes detected in clinical samples were all detected in wastewater while we also frequently detected several GI genotypes not reported in the clinical data. Application of this method into a WBE scheme, therefore, may allow for more accurate measurement of norovirus diversity within the population.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"16 4","pages":"479 - 491"},"PeriodicalIF":4.1,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09611-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142138949","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sérgio Santos-Silva, Jesús L. Romalde, Jaqueline T. Bento, Andreia V. S. Cruz, Pedro López-López, Helena M. R. Gonçalves, Wim H. M. Van der Poel, Maria S. J. Nascimento, António Rivero-Juarez, João R. Mesquita
{"title":"Serological and Molecular Survey of Hepatitis E Virus in Small Ruminants from Central Portugal","authors":"Sérgio Santos-Silva, Jesús L. Romalde, Jaqueline T. Bento, Andreia V. S. Cruz, Pedro López-López, Helena M. R. Gonçalves, Wim H. M. Van der Poel, Maria S. J. Nascimento, António Rivero-Juarez, João R. Mesquita","doi":"10.1007/s12560-024-09612-4","DOIUrl":"10.1007/s12560-024-09612-4","url":null,"abstract":"<div><p>Hepatitis E virus (HEV) is currently recognized as an emerging problem and a growing concern for public health in developed countries, with HEV infections mainly attributable to foodborne transmission of HEV-3. The zoonotic HEV genotype 3 infects a wide range of mammalian hosts, with swine considered as the primary host. This study investigates the occurrence of HEV among small ruminants in Portugal. The primary aim of the present research was to evaluate the circulation and the potential for HEV infection among sheep and goats. A total of 400 bile samples and 493 blood samples were collected from sheep and goats at a slaughterhouse in the center region of Portugal, between January 2022 and March 2023. The HEV RNA detection in bile samples was performed using a nested broad-spectrum RT-PCR targeting the ORF1 region. Serological analysis to detect anti-HEV antibodies was conducted using a commercial double-antigen sandwich multi-species ELISA. The HEV RNA was not detected in any bile samples using the nested broad-spectrum RT-PCR. Serological analysis revealed an overall HEV antibody seroprevalence of 2% (10/493, 95% CI: 0.98–3.70) among the small ruminants, namely 2.2% in goats and 2.0% in sheep. Curiously, no statistically significant association among the factors, age, sex and species and HEV seroprevalence was observed. Although HEV RNA was not detected in the bile of sheep and goats, this study the evidence of seroprevalence in these small ruminant species. Further research could provide additional insights into the factors influencing HEV transmission dynamics in small ruminants in Portugal and its potential implications for public health.</p></div>","PeriodicalId":563,"journal":{"name":"Food and Environmental Virology","volume":"16 4","pages":"516 - 524"},"PeriodicalIF":4.1,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12560-024-09612-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142131545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}