APPLICATION OF A ONE-STEP REAL TIME RT-PCR PROTOCOL FOR NOROVIRUS DETECTION IN SHELLFISH HARVERSTED AND COMMERCIALIZED IN CAMPANIA REGION

Shellfish are recognized as a potential vehicle of viral diseases. However, according to European Regulations, only bacteriological parameters (Salmonella and E. coli) are used to establish their microbiological quality. Because of the lack of a standard reference method for the detection of viruses, data on the incidence of enteric viruses in European production areas are still limited. Aim of the present study was the evaluation of the Norovirus (NoV) contamination in shellfish from different harvesting areas (both class A and B) and from registered and not- registered retails of the Campania region (Tyrrhenian Sea, Italy). Seventy shellfish (Mytilus galloprovincialis) were collected from 15 harvesting areas, one authorized and two not authorized retails and were examined for virus presence (NoV genogroups I and II) by Real Time PCR. Ten harvesting areas (66%) showed presence of viral contamination, with three of them belonging to A areas and seven to B areas. The shellfish collected from the registered retail tested positive for Norovirus presence, whereas the shellfish collected from not-authorized retails didn't show viral contamination. Twenty-nine samples were positive for genogroup II, ten of which tested positive also for NoV genogroup I

SARS-CoV-2 has been circulating in northern Italy since december 2019: evidence from environmental monitoring

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is responsible for the coronavirus disease COVID-19, a public health emergency worldwide, and Italy is among the most severely affected countries. The first autochthonous Italian case of COVID-19 was documented on February 21, 2020. We investigated the possibility that SARS-CoV-2 emerged in Italy earlier than that date, by analysing 40 composite influent wastewater samples collected - in the framework of other wastewater-based epidemiology projects - between October 2019 and February 2020 from five wastewater treatment plants (WWTPs) in three cities and regions in northern Italy (Milan/Lombardy, Turin/Piedmont and Bologna/Emilia Romagna). Twenty-four additional samples collected in the same WWTPs between September 2018 and June 2019 (i.e. long before the onset of the epidemic) were included as ‘blank’ samples. Viral concentration was performed according to the standard World Health Organization procedure for poliovirus sewage surveillance, with modifications. Molecular analysis was undertaken with both nested RT-PCR and real-rime RT-PCR assays. A total of 15 positive samples were confirmed by both methods. The earliest dates back to 18 December 2019 in Milan and Turin and 29 January 2020 in Bologna. Virus concentration in the samples ranged from below the limit of detection (LOD) to 5.6 × 104 genome copies (g.c.)/L, and most of the samples (23 out of 26) were below the limit of quantification of PCR. Our results demonstrate that SARS-CoV-2 was already circulating in northern Italy at the end of 2019. Moreover, it was circulating in different geographic regions simultaneously, which changes our previous understanding of the geographical circulation of the virus in Italy. Our study highlights the importance of environmental surveillance as an early warning system, to monitor the levels of virus circulating in the population and identify outbreaks even before cases are notified to the healthcare system

First detection of SARS-CoV-2 in untreated wastewaters in Italy

Several studies have demonstrated the advantages of environmental surveillance through the monitoring of sewage for the assessment of viruses circulating in a given community (wastewater-based epidemiology, WBE). During the COVID-19 public health emergency, many reports have described the presence of SARS-CoV-2 RNA in stools from COVID-19 patients, and a few studies reported the occurrence of SARS-CoV-2 in wastewaters worldwide. Italy is among the world's worst-affected countries in the COVID-19 pandemic, but so far there are no studies assessing the presence of SARS-CoV-2 in Italian wastewaters. To this aim, twelve influent sewage samples, collected between February and April 2020 from Wastewater Treatment Plants in Milan and Rome, were tested adapting, for concentration, the standard WHO procedure for Poliovirus surveillance. Molecular analysis was undertaken with three nested protocols, including a newly designed SARS-CoV-2 specific primer set. SARS-CoV-2 RNA detection was accomplished in volumes of 250 ml of wastewaters collected in areas of high (Milan) and low (Rome) epidemic circulation, according to clinical data. Overall, 6 out of 12 samples were positive. One of the positive results was obtained in a Milan wastewater sample collected a few days after the first notified Italian case of autochthonous SARS-CoV-2. The study confirms that WBE has the potential to be applied to SARS-CoV-2 as a sensitive tool to study spatial and temporal trends of virus circulation in the population

Análise das condições de utilização de permutadores de calor de modo a serem economicamente rentáveis

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica Perfil Energia, Refrigeração e ClimatizaçãoNeste trabalho pretende-se abordar alguns assuntos referentes à utilização de permutadores de calor em sistemas AVAC. Uma vez que os sistemas AVAC tratam o ar ambiente para depois o insuflar nos espaços climatizados, é importante abordar alguns componentes de psicrometria, como as propriedades psicrométricas e o diagrama psicrométrico. Posteriormente aborda-se a teoria dos permutadores de calor e apresenta -se os tipos mais utilizados em AVAC. A parte mais importante do trabalho consiste na criação de um algoritmo em Excel que permite, através da inserção de alguns dados, calcular a poupança anual obtida com a implementação de permutadores de calor e o tempo de amortização do investimento feito na aquisição do equipamento. Considerando o perfil de temperaturas horárias anuais de três cidades portuguesas (Bragança, Beja e Peniche), são feitos estudos,através do algoritmo, que permitem tirar algumas conclusões sobre a rentabilidade da utilização de PC em Portugal Continental. Finalmente, aborda-se um processo que possibilita uma melhoria da permuta de calor entre os caudais de ar presentes no PC e realiza-se um estudo para perceber qual o aumento da poupança anual com a aplicação do mesmo.Abstract: This work intends to address some issues concerning the use of heat exchangers in HVAC systems. It’s important to talk about psychrometric components like psychrometric properties and psychrometric chart once that the HVAC systems treat the dry air to inflate in the air-conditioning spaces. Later discusses the theory and the types of heat exchangers more used in HVAC. The most important part of this work consists in the creation of an algorithm in Excelthat allows, through the insertion of some data, calculate the annual savings achieved with the implementation of heat exchangers and the time of amortization of investment resulting from the acquisition of equipment. Considering the hourly temperature annualprofile of three Portuguese cities (Bragança, Beja e Peniche), some studies are made by means of this algorithm, which enables us to draw some conclusions about the profitability of using heat exchangers in Portugal Finally, we analyze a process that enables an improvement in the exchange of heat between the air flows inside the heat exchanger and performs a study to see the increase in annual savings due to the implementation of the process

Pepper mild mottle virus in different water matrices

Abstract Background Pepper mild mottle virus (PMMoV), a plant virus belonging to Virgoviridae, has recently been suggested as a potential viral indicator for faecal pollution in aquatic environments, since it has been found to be abundantly excreted from healthy human subjects. Methods The occurrence, amount and diversity of PMMoV was investigated in water environments by nested RT-PCR and TaqMan based quantitative PCR. During 2017-2019, 251 water samples (92 urban wastewaters, 32 treated effluents, 16 surface water samples, 9 estuarine samples, 20 seawater samples, 67 groundwater samples, and 15 drinking waters) were analysed. Results PMMoV was detected in 73/92 (79%) wastewater samples, 22/32 (69%) treated sewages, 11/16 (69%) river samples, 6/9 (67%) estuarine samples, 5/20 (25%) bathing waters, and 9/67 (13%) groundwaters, whilst drinking water samples tested always negative. Mean viral concentrations (genome copies/L) were: raw sewage 4.2 × 106, treated sewage 7.4 × 105, river 3.2 × 103, estuarine waters 9.6 × 102, seawaters 3.0 × 102, groundwaters 7.7 × 101. Conclusions This study highlights the significant occurrence of PMMoV in aquatic environment in Italy, and a clear gradient of viral prevalence and concentrations from polluted to clean waters (wastewaters to drinking waters). Key messages PMMoV is ubiquitous throughout the water cycle in Italy with different concentrations. Studies are needed to evaluate the suitability of PMMoV as a viral indicator for human fecal pollution in waters

Anesthetic Management of Pheochromocytoma Resection in Adults with Single Ventricle Physiology

Survival rates for patients with palliated congenital heart disease are increasing, and an increasing number of adults with cyanotic congenital heart disease (CCHD) might require surgical resection of pheochromocytoma-paraganglioma (PHEO-PGL). A recent study supports the idea that patients with a history of CCHD and current or historical cyanosis might be at increased risk for developing PHEO-PGL. We review the anesthetic management of two adults with single-ventricle physiology following Fontan palliation presenting for PHEO-PGL resection and review prior published case reports. We found the use of epidural analgesia to be safe and effective in the operative and postoperative management of our patients

Tracing the footprints of SARS-CoV-2 in oceanic waters

The detection of SARS-CoV-2 in water environments has predominantly focused on wastewater, neglecting its presence in oceanic waters. This study aimed to fill this knowledge gap by investigating the occurrence of SARS-CoV-2 in remote sea and oceanic waters, at large distances from the coastline. Forty-three 500-liter samples were collected between May 2022 and January 2023 from the Atlantic Ocean, the Mediterranean Sea, the Arctic region, the Persian Gulf and the Red Sea. Using molecular detection methods including real-time RT-qPCR and nested PCR followed by sequencing, we successfully detected SARS-CoV-2 RNA in 7 of the 43 marine water samples (16.3 %), and specifically in samples taken from the Atlantic Ocean and the Mediterranean Sea. The estimated concentrations of SARS-CoV-2 genome copies in the positive samples ranged from 6 to 470 per 100 l. The presence of mutations characteristic of the Omicron variant was identified in these samples by amplicon sequencing. These findings provide evidence of the unforeseen presence of SARS-CoV-2 in marine waters even at distances of miles from the coastline and in open ocean waters. It is important to consider that these findings only display the occurrence of SARS-CoV-2 RNA, and further investigations are required to assess if infectious virus can be present in the marine environment

Wastewater surveillance of SARS-CoV-2 variants in October–November 2022 in Italy: detection of XBB.1, BA.2.75 and rapid spread of the BQ.1 lineage

: This study adds insight regarding the occurrence and spread of SARS-CoV-2 Variants of Concern (VOCs) and Variants of Interest (VOIs) in Italy in October and November 2022, by testing urban wastewater collected throughout the country. A total of 332 wastewater samples were collected from 20 Italian Regions/Autonomous Provinces (APs) within the framework of national SARS-CoV-2 environmental surveillance. Of these, 164 were collected in the first week of October and 168 in the first week of November. A ∼1600 bp fragment of the spike protein was sequenced by Sanger (for individual samples) and long-read nanopore sequencing (for pooled Region/AP samples). In October, mutations characteristic of Omicron BA.4/BA.5 were detected in the vast majority (91 %) of the samples amplified by Sanger sequencing. A fraction of these sequences (9 %) also displayed the R346T mutation. Despite the low prevalence documented in clinical cases at the time of sampling, amino acid substitutions characteristic of sublineages BQ.1 or BQ.1.1 were detected in 5 % of sequenced samples from four Regions/APs. A significantly higher variability of sequences and variants was documented in November 2022, when the rate of sequences harbouring mutations of lineages BQ.1 and BQ1.1 increased to 43 %, and the number of Regions/APs positive for the new Omicron subvariant more than tripled (n = 13) compared to October. Moreover, an increase in the number of sequences with the mutation package BA.4/BA.5 + R346T (18 %), as well as the detection of variants never observed before in wastewater in Italy, such as BA.2.75 and XBB.1 (the latter in a Region where no clinical cases associated with this variant had ever been documented) was recorded. The results suggest that, as predicted by the ECDC, BQ.1/BQ.1.1 is rapidly becoming dominant in late 2022. Environmental surveillance proves to be a powerful tool for tracking the spread of SARS-CoV-2 variants/subvariants in the population