Insects and SARS-CoV-2: Analysis of the Potential Role of Vectors in European Countries.

ABSTRACT: SARS-CoV-2 is a coronavirus responsible for the pandemic that developed in China in late 2019. Transmission of the virus is predominantly direct, through exposure to infected respiratory secretions. As far as we know, arthropods play a key role in the transmission and spread of several viruses, and thus their role in the spread of COVID-19 deserves to be studied. The biological transmission of viral agents through insects is very complex. While mechanical transmission is more likely to happen, biological transmission is possible via blood-sucking arthropods, but this requires a high grade of compatibility between the vector and the pathogen. If the biological and mechanical transmission of SARS-CoV-2 by blood-sucking arthropods is excluded, a mechanical transmission by urban pests could take place. This risk is very low but it could be important in isolated environmental conditions, where other means of transmission are not possible. The presence of SARS-CoV-2 in non-blood-sucking arthropods in infected buildings, like hospitals and retirement homes, should be investigated.

A review on microbiological decontamination of fresh produce with nonthermal plasma.

Food safety is a critical public health issue for consumers and the food industry because microbiological contamination of food causes considerable social and economic burdens on health care. Most foodborne illness comes from animal production, but as of the mid-1990s in the United States and more recently in the European Union, the contribution of fresh produce to foodborne outbreaks has rapidly increased. Recent studies have suggested that sterilization with nonthermal plasma could be a viable alternative to the traditional methods for the decontamination of heat-sensitive materials or food because this technique proves capable of eliminating micro-organisms on surfaces without altering the substrate. In the last 10 years, researchers have used nonthermal plasma in a variety of food inoculated with many bacterial species. All of these experiments were conducted exclusively in a laboratory and, to our knowledge, this technique has not been used in an industrial setting. Thus, the purpose of this review is to understand whether this technology could be used at the industrial level. The latest researches using nonthermal plasma on fresh produce were analysed. These evaluations have focused on the log reduction of micro-organisms and the treatment time.

Detection of pathogenic Campylobacter, E. coli O157:H7 and Salmonella spp. in wastewater by PCR assay.

The aim of this study was the evaluation of the occurrence of pathogenic Campylobacter, Escherichia coli O157:H7, E. coli virulence genes and Salmonella spp. in different wastewater treatment plants (WWTPs) using a method based on an enrichment step and PCR. This method was sensitive enough to detect low levels (∼2 CFU100 ml(-1) of raw sewage) of all the investigated pathogens. In the WWTP samples, E. coli O157:H7 DNA and the eae gene were never found, but 33 % of influents and effluents exhibited amplicons corresponding to Shiga-like toxin I. Twenty-five percent of the influent and 8 % of the effluent exhibited the presence of Shiga-like toxin II. Campylobacter jejuni and C. coli DNA were identified in 50 and 25 % of the influents and in 8 and 25 % of the effluents, respectively. Salmonella spp. DNA was present in all the samples. Considering the results obtained, the method tested here offers a reliable and expeditious tool for evaluating the efficiency of the effluent treatment in order to mitigate contamination risk. Influent contamination by Salmonella spp. and Campylobacter spp. provides indirect information about their circulation; moreover, their presence in effluents underlines the role of WWTPs in the contamination of the receiving surface waters, which affects public health directly or indirectly.

Hospital effluents management: Chemical, physical, microbiological risks and legislation in different countries.

Hospital wastewater (HWW) can contain hazardous substances, such as pharmaceutical residues, chemical hazardous substances, pathogens and radioisotopes. Due to these substances, hospital wastewater can represent a chemical, biological and physical risk for public and environmental health. In particular, several studies demonstrate that the main effects of these substances can't be neutralised by wastewater treatment plants (WWTPs). These substances can be found in a wide range of concentrations due to the size of a hospital, the bed density, number of inpatients and outpatients, the number and the type of wards, the number and types of services, the country and the season. Some hazardous substances produced in hospital facilities have a regulatory status and are treated like waste and are disposed of accordingly (i.e., dental amalgam and medications). Legislation is quite homogeneous for these substances in all industrial countries. Problems that have emerged in the last decade concern substances and microorganisms that don't have a regulatory status, such as antibiotic residues, drugs and specific pathogens. At a global level, guidelines exist for treatment methods for these effluents, but legislation in all major industrial countries don't contain limitations on these parameters. Therefore, a monitoring system is necessary for these effluents as well as for substances and pathogens, as these elements can represent a risk to the environment and public health.

Inflammation response and cytotoxic effects in human THP-1 cells of size-fractionated PM10 extracts in a polluted urban site.

To contribute to a greater characterization of the airborne particulate matter's toxicity, size-fractionated PM10 was sampled during different seasons in a polluted urban site in Torino, a northern Italian city. Three main size fractions (PM10 - 3 µm; PM3 - 0.95 µm; PM < 0.95 µm) extracts (organic and aqueous) were assayed with THP-1 cells to evaluate their effects on cell proliferation, LDH activity, TNFα, IL-8 and CYP1A1 expression. The mean PM10 concentrations were statistically different in summer and in winter and the finest fraction PM<0.95 was always higher than the others. Size-fractionated PM10 extracts, sampled in an urban traffic meteorological-chemical station produced size-related toxicological effects in relation to season and particles extraction. The PM summer extracts induced a significant release of LDH compared to winter and produced a size-related effect, with higher values measured with PM10-3. Exposure to size-fractionated PM10 extracts did not induce significant expression of TNFα. IL-8 expression was influenced by exposure to size-fractionated PM10 extracts and statistically significant differences were found between kind of extracts for both seasons. The mean fold increases in CYP1A1 expression were statistically different in summer and in winter; winter fraction extracts produced a size-related effect, in particular for organic samples with higher values measured with PM<0.95 extracts. Our results confirm that the only measure of PM can be misleading for the assessment of air quality moreover we support efforts toward identifying potential effect-based tools (e.g. in vitro test) that could be used in the context of the different monitoring programs.

Feasibility and reliability of a questionnaire for evaluation of the exposure to indoor and outdoor air pollutants, diet and physical activity in 6-8-year-old children.

INTRODUCTION: The MAPEC-Life project aims to study the biological effects of early exposure to air pollutants on the oral mucosa cells of school-age children in five Italian cities. A questionnaire was created to evaluate the association between outdoor and indoor airborne pollutants, lifestyle, diet and biomarker effects. The feasibility and reliability of the questionnaire were evaluated. METHODS: A questionnaire was drawn up to be filled in by the parents of 6-8-year-old children. It consisted of 148 questions on the children's health, physical activity, environmental exposures and the frequency of food consumption at the main meals. First we conducted a questionnaire feasibility study involving 53 volunteer parents. We then performed a reliability study by administering the questionnaire to a further 156 parents and again one month later (test/retest method). The correlations between answers at the first and second administration of the questionnaire were evaluated using the Kappa statistic and Spearman's coefficient. RESULTS: After verifying the feasibility of the questionnaire, we conducted a reliability analysis on 132 completed questionnaires. The percentage of agreement between the first and the second responses given was over 70%, all K values being greater than 0.6. The analysis of calories and macronutrients also showed good agreement. CONCLUSIONS: The questionnaire drawn up for the study proved to be sufficiently reliable for gathering information about the factors of interest in our study of the relationship between air pollution and early biological effects in children.

PM10 in a background urban site: chemical characteristics and biological effects.

PM10 was sampled in a background urban site in Torino, a northern Italian city. PM10 extracts were tested with THP-1 and A-549 cells to evaluate their effects on cell proliferation, LDH activity, TNFα, IL8 and CYP1A1 expression, and genotoxic damage induction (Comet assay). Through Principal Component Analysis (PCA), it was observed that (1) the aqueous extracts induced the inhibition of cell proliferation in the warm season that clustered together to total ions, (2) organic extracts determined a winter cell viability reduction and (3) there was a genotoxic effect associated with PAH and metal concentrations. The analysed low PAH levels were unable to induce significant CYP1A1 expression. The results obtained confirmed that PM composition and seasonality play an important role in particle-induced toxicity. The presence of PM10-induced biological effects at a low polluted site suggested that a reduction of PM10 mass did not seem to be sufficient to reduce its toxicity.

Low-temperature, low-pressure gas plasma application on Aspergillus brasiliensis, Escherichia coli and pistachios.

AIM: The aim of this study was to investigate the effect of plasma-enhanced chemical vapour deposition (PECVD) treatment on selected bacteria and spores and to contribute to the understanding of the synergistic effect of UV-directed plasma. METHODS AND RESULTS: The experiments were conducted on pure cultures of Aspergillus brasiliensis and Escherichia coli and on naturally contaminated pistachios that were exposed to pure oxygen-, pure argon- and to a mixture of oxygen-argon-generated plasma for different treatment times and at different micro-organism concentrations. Optical emission spectroscopy (OES) measurements were performed to observe the active species in the plasma. After exposure, the effectiveness of decontamination was assessed through microbiological techniques by calculating the growth reduction on a logarithmic scale. A treatment time of 30 min resulted in a 3·5 log reduction of A. brasiliensis using pure oxygen or argon, while treatment times of 5 min, 1 min and 15 s resulted in a 5·4 log reduction using a mixture of argon and oxygen (10 : 1 v/v). Treatment times of 1 min and 30 s resulted in a 4 log reduction of E. coli with oxygen and argon, respectively, which led to a complete elimination of the micro-organisms. Two-log reductions of fungi were achieved for pistachios after a treatment time of 1 min. CONCLUSIONS: These results suggest that this newly designed plasma reactor offers good potential applications for the reduction in micro-organisms on heat-sensitive materials, such as foods. The plasma that was generated with Ar/O2 was more effective than that which was generated with pure oxygen and pure argon. SIGNIFICANCE AND IMPACT OF THE STUDY: An improvement in the knowledge about PECVD mechanisms was acquired from the chemical and biological points of view, and the suitability of the method for treating dry food surfaces was demonstrated.

Implementation of the INTERGROWTH-21st Project in Italy.

Turin, Italy, was one of the two European sites for the INTERGROWTH-21(st) Project. The sample for the Newborn Cross-Sectional Study (NCSS) was drawn from two obstetric hospitals that together account for 79% of the city's approximately 12,000 births per year. Women were recruited for the Fetal Growth Longitudinal Study (FGLS) from ten antenatal clinics serving the city's largest obstetric hospital, Azienda Ospedaliera OIRM-S. Anna. Special activities to encourage participation and raise awareness of the project in this population included obtaining an endorsement from the coordinator of the city's antenatal care service, and disseminating information about the project to women through posters and leaflets in antenatal clinics. One of the major challenges at this site was the low recruitment rate in the early phase of FGLS because of the high prevalence of smoking and of women >35 years old in the population. The addition of six extra recruiting clinics served to increase the pool of potentially eligible women who could be screened and led to a marked improvement in the recruitment rate.

Auxological evaluation of newborns.

Auxological evaluation of the newborn should be based on accurate anthropometry at birth and a reliable estimate of gestational age (GA). However, a comprehensive evaluation of the neonate should consider not only anthropometric traits at birth, but also fetal ultrasound biometry and Doppler velocimetry. Many charts have been proposed, but they are hardly comparable with each other, due to numerous methodological problems. The Italian Societies of Neonatology, of Pediatric Endocrinology and Diabetology and the Italian Society of Medical Statistics and Clinical Epidemiology promoted a multicenter survey with the aim to produce an Italian neonatal anthropometric reference (Italian Neonatal Study [INeS] charts) fulfilling the set of the criteria that a reliable neonatal chart should possess. In order to construct an international standard, an international project (INTERGROWTH-21st) has started a study aiming to create a prescriptive standard. Until an international standard is developed, the use of national updated reference charts is recommended.

Critical appraisal of different anthropometric charts to evaluate postnatal growth of preterm infants.

Preterm infants' survival has greatly increased in the last few decades thanks to the improvement in obstetrical and neonatal care. The correct evaluation of postnatal growth of these babies is nowadays of primary concern, although the definition of their optimal postnatal growth pattern is still controversial. Concerns have also been raised about the strategies to monitor their growth, specifically in relation to the charts used. At present, the charts available in clinical practice are fetal growth charts, neonatal anthropometric charts and postnatal growth charts for term infants. None of these, for different reasons, is suitable to correctly evaluate preterm infant growth. Recently, an international project has recently started aiming to create prescriptive standard for the evaluation of postnatal growth of preterm infants (INTERGROWTH-21st). Alternatively, at present, while specific charts for evaluating preterm infant postnatal growth are lacking, the best compromise is likely to be as follows: from birth to term neonatal anthropometric charts; International longitudinal charts WHO 2006 or CDC 2002 from term to childhood.

Surveillance of acute infectious gastroenteritis (1992-2009) and food-borne disease outbreaks (1996-2009) in Italy, with a focus on the Piedmont and Lombardy regions.

We describe trends in the occurrence of acute infectious gastroenteritis (1992 to 2009) and food-borne disease outbreaks (1996 to 2009) in Italy. In 2002, the Piedmont region implemented a surveillance system for early detection and control of food-borne disease outbreaks; in 2004, the Lombardy region implemented a system for surveillance of all notifiable human infectious diseases. Both systems are internet based. We compared the regional figures with the national mean using official notification data provided by the National Infectious Diseases Notification System (SIMI) and the National Institute of Statistics (ISTAT), in order to provide additional information about the epidemiology of these diseases in Italy. When compared with the national mean, data from the two regional systems showed a significant increase in notification rates of non-typhoid salmonellosis and infectious diarrhea other than non-typhoid salmonellosis, but for foodborne disease outbreaks, the increase was not statistically significant. Although the two regional systems have different objectives and structures, they showed improved sensitivity regarding notification of cases of acute infectious gastroenteritis and, to a lesser extent, food-borne disease outbreaks, and thus provide a more complete picture of the epidemiology of these diseases in Italy.

Involvement of nitro-compounds in the mutagenicity of urban Pm2.5 and Pm10 in Turin.

Fine particles can be active carriers of toxic compounds into the alveoli of the lungs. Among these compounds are numerous mutagens and carcinogens. The direct mutagenicity per unit mass of fine particulate matter (PM) is significantly higher than that of coarse particles, especially in urban areas. In this study, the mutagenic properties of urban PM2.5 and PM10 were evaluated, and the role of nitro-compounds was estimated. PM2.5 and PM10 samplings, and measurements of NOx and some PAHs were performed daily in 2007 in Turin, following a consolidated in vitro test - the Salmonella mutagenicity assay - conducted with organic extracts of PM2.5 and PM10. The mutagenic properties were assessed for each month of sampling with Salmonella typhimurium strain TA98 and TA98-derived strains: a nitroreductase-deficient mutant strain (TA98NR) and an additional nitroreductase-producing plasmid strain (YG1021). The annual measured mean levels of PM2.5 and PM10 were 34±20 and 48±18µg/m(3). The PM2.5/PM10 ratio ranged from 0.36 to 0.89. The Salmonella assay showed higher mutagenicity in autumn/winter (20±15 TA98NR; 54±39 TA98; 173±161 YG1021 net revertants/m(3)) compared with spring/summer (2±2 TA98NR; 7±8 TA98; 24±27 YG1021 net revertants/m(3)) (p<0.01). There are also statistically significant seasonal differences in the gravimetric analysis data. The number of TA98 net revertants per µg of PM2.5 is 6.5 times greater than per µg PM10. Moreover, the bioassay results showed an amplified response in the YG1021 strain and a reduced response in the TA98NR strain. The net revertant ratio TA98NR/YG1021 is 11±4 for organic extracts of PM2.5 and 13±6 for extracts of PM10 (p<0.01). There is a significant correlation between the NOx and PAH concentrations. These findings illustrate the relevant role of nitro compounds, and they underline the priority in improving preventive measures to reduce air pollution by nitrated molecules.

Endocrine disrupting activity in fruits and vegetables evaluated with the E-screen assay in relation to pesticide residues.

Food is likely to be one of the most important routes of human exposure to endocrine disrupting compounds (EDCs). In the present study, we evaluated the total estrogenic activity of fruits and vegetables, which was calculated using the human breast cancer cell line (MCF-7 BUS) proliferation assay (E-screen), in relation to pesticide residues. We analysed 44 food samples, 30 fruits and 14 vegetables. Of these samples, 10 did not contain any pesticide residues. The other 34 samples contained from 1 to 7 pesticide residues in concentrations ranging from 0.03 to 1.91 ppm. Estrogenic activity was detected in the 59% of samples tested. The positive controls used were 17-ß-estradiol (E2), the phytoestrogen genistein and the pesticide endosulfan. The average value of estradiol equivalency quantity (EEQ) for all positive samples was 0.15±0.32 µg/100g. A low correlation was found between the concentration of pesticide residues and the EEQ values (Spearman correlation r=0.376 and p=0.012). Using values obtained from the literature, we compared the estrogenic activity of food samples with the intrinsic content of phytoestrogens, but we found no correlations. Our results also suggested that the calculated intake of dietary EDCs might represent a concentration comparable to the normal endogenous estrogen concentration in human blood.

Postnatal growth of preterm infants: which reference charts?

Preterm Infants' survival has greatly increased in the last few decades thanks to the improvement in obstetrical and neonatal care. These neonates constitute the large majority of the population in neonatal intensive care units. The correct evaluation of postnatal growth of these babies is nowadays of primary concern, although the definition of their optimal postnatal growth pattern is still controversial. Concerns have also been raised about the strategies to monitor their growth,specifically in relation to the charts used. At present the available charts in clinical practice are fetal growth charts, neonatal anthropometric charts and postnatal growth charts for term infants. None of these, for different reasons, is suitable to correctly evaluate preterm infant growth. An international multicentric project has recently started a study aiming at building a prescriptive standard for the evaluation of postnatal growth of preterm infants and it will be available in the next years. At present, while an international longitudinal standard for evaluating preterm infant postnatal growth is lacking, in Italy the best compromise in clinical practice is likely to be as follows: new Italian INeS (Italian Neonatal Study) charts up to term; International longitudinal charts WHO 2006 or CDC 2002 from term to two years; finally, the Italian Society for Pediatric Endocrinology and Diabetes (SIEDP) 2006 growth charts could be suitable for monitoring the growth of these infants from two years up to 20 years of age.

Conceptual issues related to the construction of prescriptive standards for the evaluation of postnatal growth of preterm infants.

Monitoring and interpreting the growth of preterm infants is a major clinical task for neonatologists. The effectiveness of this process depends upon the robustness of the standard selected. Concerns have been raised regarding the nature of the charts currently being used, as well as their appropriateness for present-day neonatal care. To overcome these problems, there is a need for new prescriptive standards based on a population of preterm infants without evidence of impaired fetal growth and born to low-risk women followed up since early pregnancy for precise gestational age dating. Preterm infants contributing to the new standards should be free of congenital malformations and major clinical conditions associated with impaired postnatal growth. These infants should receive standardised, evidence-based clinical care and should follow current feeding recommendations based on exclusive/predominant breastfeeding. This strategy should provide a population that is conceptually as close as possible to the prescriptive approach used for the construction of the WHO infant and child growth standards. New international standards constructed in this way should contribute to the evidence-based care of these preterm infants.

Meteo-climatic conditions influence the contribution of endotoxins to PM10 in an urban polluted environment.

A decrease in inhalable particulate matter (PM10) pollution is a top priority in urban areas of northern Italy. The sources of PM10 are both anthropogenic and natural. The former have been broadly investigated while the latter are less well known. Endotoxins are natural compounds of PM10 and are potentially toxic. Endotoxins are part of the outer membrane of Gram-negative bacteria. Their health effects are linked to environmental exposure. The effects mainly consist of respiratory symptoms, including pulmonary function decline. The occurrence of endotoxins has been proven in several occupational environments where organic materials supply an optimal substrate for bacteria growth. Knowledge about the presence of these contaminants in the environment is limited. The aim of this work is to evaluate the endotoxin levels of PM10 in the urban air of Turin, and to investigate the influence of seasonal and meteo-climatic factors. The sampling was conducted from January to December 2007. Endotoxin determination was performed by an LAL assay after extraction optimization. The PM10 levels ranged from 11.90 to 104.74 microg/m(3) (48.28 +/- 23.09) while the endotoxin levels ranged between 0.09 and 0.94 EU/m(3) (0.42 +/- 0.23). The seasonal trends of PM10 and endotoxin are inversely proportional. There is a statistically significant correlation between endotoxin and temperature (r = 0.532 p < 0.01), as well as between endotoxin and relative humidity (r = -0.457 p < 0.01). However, temperature has a predominant role. We observed that urban endotoxin concentrations are narrow in range and that the contribution of endotoxins to the total PM10 is only two millionths.

Validation of SAPS 3 Admission Score and comparison with SAPS II.

BACKGROUND: The objective of this study was to validate the Simplified Acute Physiology Score SAPS 3 Admission Score (SAPS 3) and to compare its fit with that of SAPS II in an independent sample of patients admitted to a single-centre intensive care unit (ICU). METHODS: The data for all adult patients consecutively admitted to an eight-bed ICU of a 700-bed university hospital between 1 January 2006 and 2 September 2007 were collected. SAPS II and SAPS 3 were computed, as well as the predicted hospital mortality. The calibration of SAPS II and SAPS 3, according to the general equation (GE), and equations for Southern Europe and Mediterranean countries (SE&MC), and Central and Western Europe (C&WE), were assessed by the goodness-of-fit Hosmer-Lemeshow H and C statistics. Standardized mortality ratios (SMR) with 95% confidence interval (95% CI) were computed for SAPS II and SAPS 3 equations. RESULTS: Six hundred and eighty-four patients were studied (males 63%). The median age was 73 (quartiles 65-80) years. The fit of SAPS 3 using the C&WE equation (H 13.49, P=0.095; C 12.73, P=0.121) as well as that of SAPS II was acceptable (H 6.02, P=0.644; C12.08, P=0.147), while SAPS 3 GE (H 23.36, P=0.002; C 22.37, P=0.004) and S&MC (H 25.73, P=0.001; C 26.19, P=0.001) did not fit well. SAPS 3 GE, SAPS 3 SE&M Countries and the SAPS II significantly over estimated the mortality. Only 95% CI of SMR for SAPS 3 C&WE included 1 (SMR 0.97; 95% CI 0.89-1.05). CONCLUSION: Each ICU should identify the SAPS 3 equation most suitable for its case mix. The SAPS II model tended to overestimate the mortality.

Predictive value of hematological and phenotypical parameters on postchemotherapy leukocyte recovery.

BACKGROUND: Grade IV chemotherapy toxicity is defined as absolute neutrophil count <500/microL. The nadir is considered as the lowest neutrophil number following chemotherapy, and generally is not expected before the 7th day from the start of chemotherapy. The usual prophylactic dose of rHu-G-CSF (Filgrastim) is 300 microg/day, starting 24-48 h after chemotherapy until hematological recovery. However, individual patient response is largely variable, so that rHu-G-CSF doses can be different. The aim of this study was to verify if peripheral blood automated flow cytochemistry and flow cytometry analysis may be helpful in predicting the individual response and saving rHu-G-CSF. METHODS: During Grade IV neutropenia, blood counts from 30 cancer patients were analyzed daily by ADVIA 120 automated flow cytochemistry analyzer and by Facscalibur flow cytometer till the nadir. "Large unstained cells" (LUCs), myeloperoxidase index (MPXI), blasts, and various cell subpopulations in the peripheral blood were studied. At nadir rHu-G-CSF was started and 81 chemotherapy cycles were analyzed. Cycles were stratified according to their number and to two dose-levels of rHuG-CSF needed to recovery (300-600 vs. 900-1200 microg) and analyzed in relation to mean values of MPXI and mean absolute number of LUCs in the nadir phase. The linear regressions of LUCs % over time in relation to two dose-levels of rHu-G-CSF and uni-multivariate analysis of lymphocyte subpopulations, CD34(+) cells, MPXI, and blasts were also performed. RESULTS: In the nadir phase, the increase of MPXI above the upper limit of normality (>10; median 27.7), characterized a slow hematological recovery. MPXI levels were directly related to the cycle number and inversely related to the absolute number of LUCs and CD34(+)/CD45(+) cells. A faster hematological recovery was associated with a higher LUC increase per day (0.56% vs. 0.25%), higher blast (median 36.7/microL vs. 19.5/microL) and CD34(+)/CD45(+) cell (median 2.2/microL vs. 0.82/microL) counts. CONCLUSIONS: Our study showed that some biological indicators such as MPXI, LUCs, blasts, and CD34(+)/CD45(+) cells may be of clinical relevance in predicting individual hematological response to rHu-G-CSF. Special attention should be paid when nadir MPXI exceeds the upper limit of normality because the hematological recovery may be delayed.