Comparing analytical methods to detect SARS-CoV-2 in wastewater

Wastewater based epidemiology (WBE) has emerged as a reliable strategy to assess the coronavirus disease 2019 (COVID-19) pandemic. Recent publications suggest that SARS-CoV-2 detection in wastewater is technically feasible; however, many different protocols are available and most of the methods applied have not been properly validated. To this end, different procedures to concentrate and extract inactivated SARS-CoV-2 and surrogates were initially evaluated. Urban wastewater seeded with gamma-irradiated SARS-CoV-2, porcine epidemic diarrhea virus (PEDV), and mengovirus (MgV) was used to test the concentration efficiency of an aluminum-based adsorption-precipitation method and a polyethylene glycol (PEG) precipitation protocol. Moreover, two different RNA extraction methods were compared in this study: a commercial manual spin column centrifugation kit and an automated protocol based on magnetic silica beads. Overall, the evaluated concentration methods did not impact the recovery of gamma-irradiated SARS-CoV-2 nor MgV, while extraction methods showed significant differences for PEDV. Mean recovery rates of 42.9 ± 9.5%, 27.5 ± 14.3% and 9.0 ± 2.2% were obtained for gamma-irradiated SARS-CoV-2, PEDV and MgV, respectively. Limits of detection (LoD95%) for five genomic SARS-CoV-2 targets (N1, N2, gene E, IP2 and IP4) ranged from 1.56 log genome equivalents (ge)/mL (N1) to 2.22 log ge/mL (IP4) when automated system was used; while values ranging between 2.08 (N1) and 2.34 (E) log ge/mL were observed when using column-based extraction method. Different targets were also evaluated in naturally contaminated wastewater samples with 91.2%, 85.3%, 70.6%, 79.4% and 73.5% positivity, for N1, N2, E, IP2 and IP4, respectively. Our benchmarked comparison study suggests that the aluminum precipitation method coupled with the automated nucleic extraction represents a method of acceptable sensitivity to provide readily results of interest for SARS-CoV-2 WBE surveillance.The study was funded by grants from CSIC (202070E101), Generalitat Valenciana (Covid_19-SCI), MICINN co-founded by AEI/FEDER, UE (AGL2017-82909), and MICINN/AEI (PID2019-105509RJ-I00). EC-F is recipient of a predoctoral contract from the MICINN, Call 2018. WR is holder of the APOSTD/2018/150 postdoctoral contract from Generalitat Valenciana.Peer reviewe

Antiviral capacity of sanitizers against infectious viruses in process water from the produce industry under batch and continuous conditions

The presence of human enteric viruses in produce has extensively been reported. However, the significance of the quality of process water (PW) used by the produce industry and the viral inactivation capacity of water disinfection agents used to maintain the microbiological quality of PW has received limited attention. This study evaluates the antiviral disinfection efficacy of chlorine, chlorine dioxide (ClO2) and peracetic acid (PAA) at recommended operational limits in PW using hepatitis A virus (HAV), the cultivable norovirus surrogate, murine norovirus (MNV-1), and MS2 coliphages. Defined commodity representative crops (baby leaves, bell peppers, and the vegetable mix of tomatoes, cucumbers, peppers, and onions) associated with specific water-based processes were studied. Two systems classified as either batch or continuous system were used. The continuous system allows the continuously entrance of sanitizer solution and organic matter added to the washing tank to simulate the conditions of an industry wash tank. Batch scale experiments showed that 20 mg/L chlorine and 3 mg/L chlorine dioxide completely inactivated MNV-1 and MS2 (mean of 5 log) after 1 min contact time regardless of the PW type. However, the infectivity of HAV was reduced only by less than 2 log after 1 min for chlorine and chlorine dioxide and the complete inactivation was not observed even after 10 min. On the contrary, residual viral infectivity/viability of HAV, MNV-1 and MS2 was observed for PAA in the three types of PW. The inactivation kinetic models for MS2 coliphages were developed based on the data obtained under the continuous system comparing the three types of PW. Chlorine (5 mg/L) and chlorine dioxide (2-3 mg/L) avoided the accumulation of MS2 below the detection limit while PAA (80 mg/L) was unable to prevent it independently of the type of PW. In summary, in the washing operation, it is a key objective to reach virus inactivation through the selection of the most effective sanitizer by guaranteeing that sufficient concentration and contact times prevent the risk of viral cross-contamination

Wastewater based epidemiology beyond SARS-CoV-2: Spanish wastewater reveals the current spread of Monkeypox virus

Besides nasopharyngeal swabs, monkeypox virus (MPXV) DNA has been detected in a variety of samples such as saliva, semen, urine and fecal samples. Using the environmental surveillance network previously developed in Spain for the routine wastewater surveillance of SARS-CoV-2 (VATar COVID-19), we have analyzed the presence of MPXV DNA in wastewater from different areas of Spain. Samples (n=312) from 24 different wastewater treatment plants were obtained between May 9 (week 22_19) and August 4 (week 22_31), 2022. Following concentration of viral particles by flocculation, a qPCR procedure allowed us to detect MPXV DNA in 63 wastewater samples collected from May 16 to August 4, 2022, with values ranging between 2.2 per 103 to 8.7 per 104 genome copies (gc)/L. This study shows that MPXV DNA can be reproducibly detected by qPCR in longitudinal samples collected from different Spanish wastewater treatment plants. According to data from the National Epidemiological Surveillance Network (RENAVE) in Spain a total of 6,119 cases have been confirmed as of August 19, 2022. However, and based on the wastewater data, the reported clinical cases seem to be underestimated and asymptomatic infections may be more frequent than expected.his research was supported by the European Commission NextGenerationEU fund, through CSIC's Global Health Platform (PTI Salud Global) and samples were obtained from the COVID-19 wastewater surveillance project (VATar COVID-19) funded by the Spanish Ministry for the Ecological Transition and the Demographic Challenge and the Spanish Ministry of Health. IGG is recipient of a predoctoral contract from the Generalitat Valenciana (ACIF/2021/181) and AP-C was supported by a postdoctoral fellowship (APOSTD/2021/292). PT is holding a Ramon y Cajal contract from the Ministerio de Ciencia e Innovacion and AC is recipient of a predoctoral contract FI-SDUR from the Generalitat de CatalunyaN

Spanish wastewater reveals the current spread of Monkeypox virus

Besides nasopharyngeal swabs, monkeypox virus (MPXV) DNA has been detected in a variety of samples such as saliva, semen, urine and fecal samples. Using the environmental surveillance network previously developed in Spain for the routine wastewater surveillance of SARS-CoV-2 (VATar COVID-19), we have analyzed the presence of MPXV DNA in wastewater from different areas of Spain. Samples (n = 312) from 24 different wastewater treatment plants were obtained between May 9 (week 19 of 2022) and August 4 (week 31 of 2022). Following concentration of viral particles by a validated aluminum adsorption-precipitation method, a qPCR procedure allowed us to detect MPXV DNA in 56 wastewater samples collected from May 16 to August 4, 2022, with values ranging between 2.2 × 103 to 8.7 × 104 genome copies (gc)/L. This study shows that MPXV DNA can be reproducibly detected by qPCR in longitudinal samples collected from different Spanish wastewater treatment plants. According to data from the National Epidemiological Surveillance Network (RENAVE) in Spain a total of 6,119 cases have been confirmed as of August 19, 2022. However, and based on the wastewater data, the reported clinical cases seem to be underestimated and asymptomatic infections may be more frequent than expected.This research was supported by the European Commission NextGenerationEU fund, through CSIC's Global Health Platform (PTI Salud Global), project CEX2021-001189-S MCIN/AEI / 10.13039/501100011033 and Fundación Séneca (Region of Murcia). Samples were obtained from the COVID-19 wastewater surveillance project (VATar COVID-19) funded by the Spanish Ministry for the Ecological Transition and the Demographic Challenge and the Spanish Ministry of Health.With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2021-001189-S).Peer reviewe

Food Safety

24 Páginas.-- 2 FigurasEmerging and re-emerging food safety risks are driven by climate change and other factors like globalization, water scarcity, population aging, and migration, among others. These factors affect the persistence and occurrence of conventional and emerging primary risks. Successful initiatives to manage emerging risks need to identify, evaluate, and prioritize potential risks as well as to respond with sustainable strategies able to reduce the threatsPeer reviewe

Hispanic Latin America, Spain and the Spanish-speaking Caribbean: A rich source of reference material for public health, epidemiology and tropical medicine

There is a multiplicity of journals originating in Spain and the Spanish-speaking countries of Latin America and the Caribbean (SSLAC) in the health sciences of relevance to the fields of epidemiology and public health. While the subject matter of epidemiology in Spain shares many features with its neighbours in Western Europe, many aspects of epidemiology in Latin America are particular to that region. There are also distinctive theoretical and philosophical approaches to the study of epidemiology and public health arising from traditions such as the Latin American social medicine movement, of which there may be limited awareness. A number of online bibliographic databases are available which focus primarily on health sciences literature arising in Spain and Latin America, the most prominent being Literatura Latinoamericana en Ciencias de la Salud (LILACS) and LATINDEX. Some such as LILACS also extensively index grey literature. As well as in Spanish, interfaces are provided in English and Portuguese. Abstracts of articles may also be provided in English with an increasing number of journals beginning to publish entire articles written in English. Free full text articles are becoming accessible, one of the most comprehensive sources being the Scientific Electronic Library Online (SciELO). There is thus an extensive range of literature originating in Spain and SSLAC freely identifiable and often accessible online, and with the potential to provide useful inputs to the study of epidemiology and public health provided that any reluctance to explore these resources can be overcome. In this article we provide an introduction to such resources

Maximum levels of cross‐contamination for 24 antimicrobial active substances in non‐target feed. Part 12: Tetracyclines: tetracycline, chlortetracycline, oxytetracycline, and doxycycline

[EN] The specific concentrations of tetracycline, chlortetracycline, oxytetracycline and doxycycline in non-target feed for food-producing animals, below which there would not be an effect on the emergence of, and/or selection for, resistance in bacteria relevant for human and animal health, as well as the specific antimicrobial concentrations in feed which have an effect in terms of growth promotion/increased yield were assessed by EFSA in collaboration with EMA. Details of the methodology used for this assessment, associated data gaps and uncertainties are presented in a separate document. To address antimicrobial resistance, the Feed Antimicrobial Resistance Selection Concentration (FARSC) model developed specifically for the assessment was applied. The FARSC for these four tetracyclines was estimated. To address growth promotion, data from scientific publications obtained from an extensive literature review were used. Levels in feed that showed to have an effect on growth promotion/increased yield were reported for tetracycline, chlortetracycline, oxytetracycline, whilst for doxycycline no suitable data for the assessment were available. Uncertainties and data gaps associated with the levels reported were addressed. It was recommended to perform further studies to supply more diverse and complete data related to the requirements for calculation of the FARSC for these antimicrobialsSIThe specific concentrations of tetracycline, chlortetracycline, oxytetracycline and doxycycline in non-target feed for food-producing animals, below which there would not be an effect on the emergence of, and/or selection for, resistance in bacteria relevant for human and animal health, as well as the specific antimicrobial concentrations in feed which have an effect in terms of growth promotion/increased yield were assessed by EFSA in collaboration with EMA. Details of the methodology used for this assessment, associated data gaps and uncertainties are presented in a separate document. To address antimicrobial resistance, the Feed Antimicrobial Resistance Selection Concentration (FARSC) model developed specifically for the assessment was applied. The FARSC for these four tetracyclines was estimated. To address growth promotion, data from scientific publications obtained from an extensive literature review were used. Levels in feed that showed to have an effect on growth promotion/increased yield were reported for tetracycline, chlortetracycline, oxytetracycline, whilst for doxycycline no suitable data for the assessment were available. Uncertainties and data gaps associated with the levels reported were addressed. It was recommended to perform further studies to supply more diverse and complete data related to the requirements for calculation of the FARSC for these antimicrobial

Maximum levels of cross‐contamination for 24 antimicrobial active substances in non‐target feed. Part 11: Sulfonamides

[EN] The specific concentrations of sulfonamides in non-target feed for food-producing animals, below which there would not be an effect on the emergence of, and/or selection for, resistance in bacteria relevant for human and animal health, as well as the specific antimicrobial concentrations in feed which have an effect in terms of growth promotion/increased yield were assessed by EFSA in collaboration with EMA. Details of the methodology used for this assessment, associated data gaps and uncertainties, are presented in a separate document. To address antimicrobial resistance, the Feed Antimicrobial Resistance Selection Concentration (FARSC) model developed specifically for the assessment was applied. However, due to the lack of data on the parameters required to calculate the FARSC, it was not possible to conclude the assessment until further experimental data are available. To address growth promotion, data from scientific publications obtained from an extensive literature review were used. Levels in feed that showed to have an effect on growth promotion/increased yield were identified for three sulfonamides: sulfamethazine, sulfathiazole and sulfamerazine. It was recommended to carry out studies to generate the data that are required to fill the gaps which prevented the calculation of the FARSC for these antimicrobials.SIThe BIOHAZ Panel, leading Panel in charge of the adoption of the scientificopinion and assessment of Term of Reference 1 (ToR1, antimicrobial resistance) wishes to thank thefollowing for the support provided to this scientific output: EFSA Panel on Animal Health and Welfare(AHAW Panel), who supported ToR1 assessments development and endorsement of those sectionsunder their remit (animal production, main use of antimicrobials); EFSA Panel for Additives andProducts or Substances used in Animal Feed (FEEDAP), in charge of the assessment and endorsementof ToR2, and providing advice and data needed for ToR1 assessments; European Medicines Agency(EMA), who was represented by an external expert and EMA secretariat as members of the WorkingGroup (WG); Valeria Bortolaia, who was member of the WG until 17 April 2020; EFSA staff members:Angelica Amaduzzi, Gina Cioacata, Pilar Garc ıa-Vello, Michaela Hempen, Rita Navarrete, Daniel Plazaand Anita Radovnikovic; EMA staff members: Barbara Freischem, Zoltan Kunsagi, Nicholas Jarrett, JordiTorren, and Julia F abrega (currently EFSA staff). The BIOHAZ Panel wishes also to acknowledge theEMA Committee for Medicinal Products for Veterinary Use (CVMP) and their experts

Maximum levels of cross‐contamination for 24 antimicrobial active substances in non‐target feed. Part 2: Aminoglycosides/aminocyclitols: apramycin, paromomycin, neomycin and spectinomycin

[EN] The specific concentrations of apramycin, paromomycin, neomycin and spectinomycin in non-target feed for food-producing animals, below which there would not be an effect on the emergence of, and/or selection for, resistance in bacteria relevant for human and animal health, as well as the specific antimicrobial concentrations in feed which have an effect in terms of growth promotion/increased yield, were assessed by EFSA in collaboration with EMA. Details of the methodology used for this assessment, associated data gaps and uncertainties, are presented in a separate document. To address antimicrobial resistance, the Feed Antimicrobial Resistance Selection Concentration (FARSC) model developed specifically for the assessment was applied. However, due to the lack of data on the parameters required to calculate the FARSC for these antimicrobials, it was not possible to conclude the assessment until further experimental data become available. To address growth promotion, data from scientific publications obtained from an extensive literature review were used. Levels in feed that showed to have an effect on growth promotion/increased yield were reported for apramycin and neomycin, whilst for paromomycin and spectinomycin, no suitable data for the assessment were available. It was recommended to carry out studies to generate the data that are required to fill the gaps which prevented the calculation of the FARSC for these four antimicrobialsSIThe BIOHAZ Panel, leading Panel in charge of the adoption of the scientific opinion and assessment of Term of Reference 1 (ToR1, antimicrobial resistance) wishes to thank the following for the support provided to this scientific output: EFSA Panel on Animal Health and Welfare (AHAW Panel), who supported ToR1 assessments development and endorsement of those sections under their remit (animal production, main use of antimicrobials); EFSA Panel for Additives and Products or Substances used in Animal Feed (FEEDAP), in charge of the assessment and endorsement of ToR2, and providing advice and data needed for ToR1 assessments; European Medicines Agency (EMA), who was represented by an external expert and EMA secretariat as members of the Working Group (WG); Valeria Bortolaia, who was member of the WG until 17 April 2020; EFSA staff members: Angelica Amaduzzi, Gina Cioacata, Pilar Garc ıa-Vello, Michaela Hempen, Rita Navarrete, Daniel Plaza and Anita Radovnikovic; EMA staff members: Barbara Freischem, Zoltan Kunsagi, Nicholas Jarrett, Jordi Torren, and Julia F abrega (currently EFSA staff). The BIOHAZ Panel wishes also to acknowledge the EMA Committee for Medicinal Products for Veterinary Use (CVMP) expert