S100B Affects Gut Microbiota Biodiversity.

This in vivo study in mice addresses the relationship between the biodiversity of the microbiota and the levels of S100B, a protein present in enteroglial cells, but also in foods such as milk. A positive significant correlation was observed between S100B levels and Shannon values, which was reduced after treatment with Pentamidine, an inhibitor of S100B function, indicating that the correlation was influenced by the modulation of S100B activity. Using the bootstrap average method based on the distribution of the S100B concentration, three groups were identified, exhibiting a significant difference between the microbial profiles. Operational taxonomic units, when analyzed by SIMPER analysis, showed that genera regarded to be eubiotic were mainly concentrated in the intermediate group, while genera potentially harboring pathobionts often appeared to be more concentrated in groups where the S100B amounts were very low or high. Finally, in a pilot experiment, S100B was administered orally, and the microbial profiles appeared to be modified accordingly. These data may open novel perspectives involving the possibility of S100B-mediated regulation in the intestinal microbiota.

Metagenomic analysis of bacterial community in a travertine depositing hot spring.

Several factors influence bacteria biodiversity in hot springs. The impact of biotic and abiotic pathways on travertine deposition plays a key role in microbial ecology and in the final composition of the waterborne microbiota. The metabolism of some bacterial groups such as photoautotrophs or lithoautotrophs influences water chemistry, favoring carbonate precipitation processes. The role of microbial mats in mineral precipitation processes is not fully clarified. For the first time, a comprehensive metagenomic analysis has been undertaken in the historical Bullicame hot spring. Bacterial biodiversity was characterized and biomineralization activities were assigned to different genera. A higher biodiversity in mat samples compared to water samples was observed: Shannon index of 3.34 and 0.86, respectively. Based on the functional assignment of each Operational Taxonomic Unit, the bacteria involved in biologically- induced mineralization are prevalent in mat and released in the water. According to the principle that each geothermal water specimen has distinctive physic-chemical characteristics, our results suggest new interacting bio-actions within these ecosystems. The saturation index and the chemical composition, as the high concentration of sulfur species and HCO3, can be linked to create a selective environment where pioneer communities are able to live and shape the ecosystem.

[Some considerations on revision of legislation Hygiene-sanitary for the management of swimming pools for swimming-pool use and the National Consultation of the Ministry of Health]. / Alcune considerazioni sulla revisione della normativa igienico-sanitaria per la gestione di piscine ad uso natatorio e la Consultazione Nazionale del Ministero della Salute.

After over a decade the Italian regulation on hygiene and surveillance in swimming pools is under revision based on WHO guidelines and current laws from other European countries. The Ministry of Health is proposing an update of the Annex 1 that contains physical, chemical and microbiological indicators for monitoring swimming pool hygiene. The authors review current regulations and the new proposal for updating surveillance. A comparison of Annex 1 as approved in 2003 and the new version as proposed in 2016 allowed to enlighten innovative improvements. The suggested updates underwent a public consultation through an online questionnaire involving citizens and stakeholders, following the required procedure for accessing the final approval of a new regulation. The updates include new requirements and changes involving several controls and microbiological parameters. Safety of recreational waters is presently under revision in different European countries. This updating process is a valuable opportunity to implement critical control points and improve monitoring. The final aim is an optimization of surveillance in terms of efficacy and costs, with an advantage for both the National Health Service as the swimming pool management.

Infection control in healthcare settings: perspectives for mfDNA analysis in monitoring sanitation procedures.

BACKGROUND: Appropriate sanitation procedures and monitoring of their actual efficacy represent critical points for improving hygiene and reducing the risk of healthcare-associated infections. Presently, surveillance is based on traditional protocols and classical microbiology. Innovation in monitoring is required not only to enhance safety or speed up controls but also to prevent cross infections due to novel or uncultivable pathogens. In order to improve surveillance monitoring, we propose that biological fluid microflora (mf) on reprocessed devices is a potential indicator of sanitation failure, when tested by an mfDNA-based approach. The survey focused on oral microflora traces in dental care settings. METHODS: Experimental tests (n = 48) and an "in field" trial (n = 83) were performed on dental instruments. Conventional microbiology and amplification of bacterial genes by multiple real-time PCR were applied to detect traces of salivary microflora. Six different sanitation protocols were considered. A monitoring protocol was developed and performance of the mfDNA assay was evaluated by sensitivity and specificity. RESULTS: Contaminated samples resulted positive for saliva traces by the proposed approach (CT < 35). In accordance with guidelines, only fully sanitized samples were considered negative (100 %). Culture-based tests confirmed disinfectant efficacy, but failed in detecting incomplete sanitation. The method provided sensitivity and specificity over 95 %. CONCLUSIONS: The principle of detecting biological fluids by mfDNA analysis seems promising for monitoring the effectiveness of instrument reprocessing. The molecular approach is simple, fast and can provide a valid support for surveillance in dental care or other hospital settings.

Vitamin D status and COVID-19 prevention in a worker subgroup in Italy.

BACKGROUND: Low levels of vitamin D are widespread in the world's population and associated with sun exposure, genetics, and lifestyles. Office workers in different occupational sectors seem more vulnerable than others. Scientific evidence reports a contribution of vitamin D in resistance to infections, opening to supplementation as a preventive action against pathogens, including SARS-CoV-2. OBJECTIVE: A pilot campaign in the workplace during the coronavirus 2019 (COVID-19) pandemic was conducted based on the preliminary measurement of vitamin D amount and its integration. METHODS: A preventive action to contrast the deficiency of vitamin D was offered to a population of 700 bank employees. Vitamin D supplementation was performed between April and June 2021, on workers (n = 139) and showed 25(OH)D serum levels ≤ 30 ng/ml. Demographic, anthropometric and lifestyle information were collected by survey and changes in the serum 25(OH)D amounts were monitored. RESULTS: The adherence of the target population to the prevention campaign was 21%. 75% of the enrolled workers had low levels of vitamin D. After the intervention, serum vitamin D levels increased (1.28-fold;p = 0.0001) and 80% of the subjects reported optimal values > 30 ng/ml. Only 2.9% reported slight flu-like symptoms, but only 0.7% was confirmed as COVID-19, with respect to a ten-fold higher incidence in the general population. CONCLUSIONS: Vitamin D supplementation can be achieved by simple and noninvasive approaches and can bring along further insights into health literacy on diet and lifestyles, representing an opportunity to protect the population by the widespread state of vitamin deficiency.

Antimicrobial Effectiveness of Innovative Photocatalysts: A Review.

Waterborne pathogens represent one of the most widespread environmental concerns. Conventional disinfection methods, including chlorination and UV, pose several operational and environmental problems; namely, formation of potentially hazardous disinfection by-products (DBPs) and high energy consumption. Therefore, there is high demand for effective, low-cost disinfection treatments. Among advanced oxidation processes, the photocatalytic process, a form of green technology, is becoming increasingly attractive. A systematic review was carried out on the synthesis, characterization, toxicity, and antimicrobial performance of innovative engineered photocatalysts. In recent decades, various engineered photocatalysts have been developed to overcome the limits of conventional photocatalysts using different synthesis methods, and these are discussed together with the main parameters influencing the process behaviors. The potential environmental risks of engineered photocatalysts are also addressed, considering the toxicity effects presented in the literature.

The role of vitamin D in prevention OF COVID-19 and its severity: an umbrella review.

BACKGROUND AND AIM: Vitamin D is known to modulate immune response and its deficiency was associated with respiratory distress in patients hospitalized for pneumonia. Nevertheless, numerous reviews on vitamin D in COVID-19 patients have shown conflicting results, as previously reported also for other respiratory diseases (e.g., influenza). METHODS: This umbrella review aims to assess whether low serum 25-OHD is associated with susceptibility to COVID 19, their severity, and mortality. A total of 1559 studies were excluded after the title, abstract and full-text articles screening and 9 papers were included in this review: 2 systematic reviews and 7 metanalysis. RESULTS: The findings of this review that summarized studies from 5 WHO regions (European Region, Region of the Americas, South-East Asia Region, Eastern Mediterranean Region, Western Pacific Region) to exclusion only African region, show that low serum 25-OHD levels are associated with higher infection risks for COVID-19. CONCLUSIONS: Although the umbrella findings indicate a potential role of vitamin D deficiency in COVID-19 severity in hospitalized patients and showing an association between Vitamin D supplementation and COVID-19 severity, however, more robust data from randomized controlled trials are further needed to confirm a possible association with the mortality rates.

Photocatalytic Treatments for Personal Protective Equipment: Experimental Microbiological Investigations and Perspectives for the Enhancement of Antimicrobial Activity by Micrometric TiO2.

The COVID-19 pandemic has led to countries enforcing the use of facial masks to prevent contagion. However, acquisition, reuse, and disposal of personal protective equipment (PPE) has generated problems, in regard to the safety of individuals and environmental sustainability. Effective strategies to reprocess and disinfect PPE are needed to improve the efficacy and durability of this equipment and to reduce waste load. Thus, the addition of photocatalytic materials to these materials, combined with light exposure at specific wavelengths, may represent promising solutions. To this aim, we prepared a series of masks by depositing micrometer-sized TiO2 on the external surfaces; the masks were then contaminated with droplets of bacteria suspensions and the coatings were activated by light radiation at different wavelengths. A significant reduction in the microbial load (over 90%, p < 0.01) was observed using both Gram negative (E. coli) and Gram positive (S. aureus) bacteria within 15 min of irradiation, with UV or visible light, including sunlight or artificial sources. Our results support the need for further investigations on self-disinfecting masks and other disposable PPE, which could positively impact (i) the safety of operators/workers, and (ii) environmental sustainability in different occupational or recreational settings.

Monitoring COVID-19 Transmission Risks by Quantitative Real-Time PCR Tracing of Droplets in Hospital and Living Environments.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) environmental contamination occurs through droplets and biological fluids released in the surroundings from patients or asymptomatic carriers. Surfaces and objects contaminated by saliva or nose secretions represent a risk for indirect transmission of coronavirus disease 2019 (COVID-19). We assayed surfaces from hospital and living spaces to identify the presence of viral RNA and the spread of fomites in the environment. Anthropic contamination by droplets and biological fluids was monitored by detecting the microbiota signature using multiplex quantitative real-time PCR (qPCR) on selected species and massive sequencing on 16S amplicons. A total of 92 samples (flocked swabs) were collected from critical areas during the pandemic, including indoor (three hospitals and three public buildings) and outdoor surfaces exposed to anthropic contamination (handles and handrails, playgrounds). Traces of biological fluids were frequently detected in spaces open to the public and on objects that are touched with the hands (>80%). However, viral RNA was not detected in hospital wards or other indoor and outdoor surfaces either in the air system of a COVID hospital but only in the surroundings of an infected patient, in consistent association with droplet traces and fomites. Handled objects accumulated the highest level of multiple contaminations by saliva, nose secretions, and fecal traces, further supporting the priority role of handwashing in prevention. In conclusion, anthropic contamination by droplets and biological fluids is widespread in spaces open to the public and can be traced by qPCR. Monitoring fomites can support evaluation of indirect transmission risks for coronavirus or other flu-like viruses in the environment.IMPORTANCE Several studies have evaluated the presence of SARS-CoV-2 in the environment. Saliva and nasopharyngeal droplets can land on objects and surfaces, creating fomites. A suitable indicator would allow the detection of droplets or biofluids carrying the virus. Therefore, we searched for viral RNA and droplets and fomites on at risk surfaces. We monitored by qPCR or next generation sequencing (NGS) droplets through their microbiota. Although the study was performed during the pandemic, SARS-CoV-2 was not significantly found on surfaces, with the only exception of environmental areas near infectious patients. Conversely, anthropic contamination was frequent, suggesting a role for biofluids as putative markers of indirect transmission and risk assessment. Moreover, all SARS-CoV-2-contaminated surfaces showed droplets' microbiota. Fomite monitoring by qPCR may have an impact on public health strategies, supporting prevention of indirect transmission similarly to what is done for other communicable diseases (e.g., influenza and influenza-like infections).

The microbiota as a candidate biomarker for SPA pools and SPA thermal spring stability after seismic events.

Worldwide, the location of thermal springs overlaps seismic areas, and the higher occurrence of earthquakes may impact on water stability and safety. The hydrogeological perturbations pose environmental and public health risks that can be monitored by well-established chemical, physical and biological parameters. Specific health concerns involve the exposure of the population to the medical or wellness uses of SPA thermal waters, e.g. in respiratory or hydropinic treatments as well as during rehabilitative or recreational activities in pools. Since SPA waters are characterized by their own microbiota, we analysed by 16S amplicon sequencing the dynamics of water microbial communities after the August 2017 Ischia island earthquake. For the first time, we report the impact of a seismic event on a thermal spring water, whose microbiota was deeply characterized before and immediately after the natural disaster. The biodiversity stability of the water underwent a dramatic disturbance following the earthquake, as summarized by a Shannon index moving from 1.300 during May 2016-July 2017, up to 1.600 during the first 20-70 h after the event and slightly slowing down to 1.500 after 30 days and to 1.400 after 6 months. Microbiota analysis showed a sudden reduction of the relative abundance of autochthone thermophilic species within the first 20 h and a parallel increase of other thermophilic species as well as of ectopic bacteria from soil, sediments, sea, freshwater and wastewaters. Cultivable mesophilic bacteria were observed only in the first 20 h sample (7 × 103/L), even if the presence of faecal contamination traces was detected by Real Time PCR also up to 70 h after the disaster. OTUs analysis of putative metabolic functions showed several changes between pre and post event, such as in the distribution of Sulphur metabolizing and Carbon fixation species. The restoration of the original pattern followed a slow trend, requiring over six months. The observed results confirm the impact of the earthquake on the microbiota structure of the underground thermal spring water, suggesting further perspectives for monitoring water stability and safety issues by a metagenomic approach.

Mediterranean Diet, Physical Activity and Gut Microbiome Composition: A Cross-Sectional Study among Healthy Young Italian Adults.

BACKGROUND: This cross-sectional study aimed to explore the microbial composition of the gut and its possible association with the Mediterranean diet (MD) after adjusting for demographic and anthropometric characteristics in a sample of healthy young Italian adults. METHODS: Gut microbiota, demographic information, and data on adherence to MD and physical activity (PA) habits were collected in a sample of 140 university students (48.6% males, mean age 22.5 ± 2.9) with a mean body mass index (BMI) of 22.4 ± 2.8 kg/m2 (15.2-33.8) and a mean PA level of 3006.2 ± 2973.6 metabolic equivalent (MET)-minutes/week (148-21,090). RESULTS: A high prevalence of Firmicutes and Bacteroidetes was found in all the fecal samples. Significant dissimilarities in the microbiota composition were found on the basis of MD adherence and PA levels (p = 0.001). At the genus level, Streptococcus and Dorea were highly abundant in overweight/obese individuals, Ruminococcus and Oscillospira in participants with lower adherence to MD, and Lachnobacterium in subjects with low levels of PA (p = 0.001). A significantly higher abundance of Paraprevotella was shown by individuals with lower BMI, lower MD adherence, and lower PA levels (p = 0.001). CONCLUSIONS: This study contributes to the characterization of the gut microbiome of healthy humans. The findings suggest the role of diet and PA in determining gut microbiota variability.

Serum S100B protein as a marker of severity in Covid-19 patients.

SARS-CoV-2 infection shows a wide-ranging clinical severity, requiring prognostic markers. We focused on S100B, a calcium-binding protein present in biological fluids, being a reliable biomarker in disorders having inflammatory processes as common basis and RAGE as main receptor. Since Covid-19 is characterized by a potent inflammatory response also involving RAGE, we tested if S100B serum levels were related to disease severity. Serum samples (n = 74) were collected from hospitalized SARS-CoV-2 positive patients admitted to Covid center. Illness severity was established by admission clinical criteria and Covid risk score. Treatment protocols followed WHO guidelines available at the time. Circulating S100B was determined by ELISA assay. Statistical analysis used Pearson's χ2 test, t-Test, and ANOVA, ANCOVA, Linear Regression. S100B was detected in serum from Covid-19 patients, significantly correlating with disease severity as shown both by the level of intensity of care (p < 0.006) as well by the value of Covid score (Multiple R-squared: 0.3751); the correlation between Covid-Score and S100B was 0.61 (p < 0.01). S100B concentration was associated with inflammation markers (Ferritin, C-Reactive Protein, Procalcitonin), and organ damage markers (Alanine Aminotransferase, Creatinine). Serum S100B plays a role in Covid-19 and can represent a marker of clinical severity in Sars-CoV-2 infected patients.

Untouchability of natural spa waters: Perspectives for treatments within a personalized water safety plan.

Natural SPA waters and their environments were known since ancient times and used for health or recreational purposes in different societies, worldwide. The composition and uses of these spring waters may not allow standard disinfection in pools, representing a challenge for hygiene management. Several safety and quality procedures were proposed, but a systematic approach is still needed. Here, we focus on alternative strategies to provide hints for developing a sustainable Water Safety Plan, based on intrinsic water properties and photocatalytic materials. The antimicrobial activity of four different SPA waters with high mineral content and one drinkable spring water with a low mineral content, was assessed and then tested for the additional bactericidal activity of Titanium Dioxide (TiO2) nanomaterials and/or light exposure at different wavelengths (200-635 nm). A native antibacterial activity was observed in all high mineral content waters, with a CFU reduction of 75-80%. The bactericidal action of TiO2 showed an additional incremental effect, with a reduction of over 99% within 2-5 h. Interestingly, the antibacterial photocatalytic effect was detected also in the visible light range, with a possible pick around 450-455 nm, blue-light. Based on observed results, we propose a model for developing a water safety plan, considering water properties and bather exposure. This candidate approach is personalized on water composition and pool use, trying to avoid chemical disinfectants. Photocatalytic nanotechnologies represent one of the promising alternative treatments and can provide novel perspectives for a sustainable managing of natural SPA water hygiene.

Artificial-turf surfaces for sport and recreational activities: microbiota analysis and 16S sequencing signature of synthetic vs natural soccer fields.

Synthetic fibres are used in place of the natural grass worldwide, for realizing playgrounds, soccer fields and even domestic gardens or recreational structures. An intensive use of artificial turf is currently observed in sports facilities, due to lower costs, higher sustainability in recycling of materials, and advantages related to athletic practice and performance. However, even if chemical and physical risks were studied, the microbiological component was not fully addressed, especially considering a comprehensive evaluation of the microbiota in synthetic vs natural playground surfaces. Here, we investigated the microbial community present on soccer fields, using Next Generation Sequencing and a 16S amplicon sequencing approach. Artificial and natural turfs show own ecosystems with different microbial profiles and a mean Shannon's diversity value of 2.176 and 2.475, respectively. The bacterial community is significantly different between facilities (ANOSIM: R = 0.179; p < 0.001) and surface materials (ANOSIM: R = 0.172; p < 0.005). The relative abundance of potentially pathogenic bacterial OTUs was higher in synthetic than in natural samples (ANOVA, F = 2.2). Soccer fields are characterized by their own microbiota, showing a different 16S amplicon sequencing signature between natural and artificial turfs.

Potential testing of reprocessing procedures by real-time polymerase chain reaction: A multicenter study of colonoscopy devices.

BACKGROUND: Reprocessing of endoscopes is key to preventing cross-infection after colonoscopy. Culture-based methods are recommended for monitoring, but alternative and rapid approaches are needed to improve surveillance and reduce turnover times. A molecular strategy based on detection of residual traces from gut microbiota was developed and tested using a multicenter survey. METHODS: A simplified sampling and DNA extraction protocol using nylon-tipped flocked swabs was optimized. A multiplex real-time polymerase chain reaction (PCR) test was developed that targeted 6 bacteria genes that were amplified in 3 mixes. The method was validated by interlaboratory tests involving 5 reference laboratories. Colonoscopy devices (n = 111) were sampled in 10 Italian hospitals. Culture-based microbiology and metagenomic tests were performed to verify PCR data. RESULTS: The sampling method was easily applied in all 10 endoscopy units and the optimized DNA extraction and amplification protocol was successfully performed by all of the involved laboratories. This PCR-based method allowed identification of both contaminated (n = 59) and fully reprocessed endoscopes (n = 52) with high sensibility (98%) and specificity (98%), within 3-4 hours, in contrast to the 24-72 hours needed for a classic microbiology test. Results were confirmed by next-generation sequencing and classic microbiology. CONCLUSIONS: A novel approach for monitoring reprocessing of colonoscopy devices was developed and successfully applied in a multicenter survey. The general principle of tracing biological fluids through microflora DNA amplification was successfully applied and may represent a promising approach for hospital hygiene.

Molecular identification of vaginal fluid by microbial signature.

The discrimination of body fluids in forensic examinations can play an important role in crime scene reconstruction. Conventional methods rely on the detection of antigens or enzymatic activity, limiting detection sensitivity and specificity, particularly on old forensic samples. Methods based on human RNA analysis are not easily applicable to samples exposed to harsh and degrading environments. An alternative approach based on the identification of prokaryotic genomes was developed. Specific bacterial communities are characteristic typical of different human non-sterile body fluids: the molecular characterization of a microbial signature, and not the typing of single bacterial species, can effectively lead to univocal identification of these fluids. A multiplex real time PCR assay was developed using oligonucleotide mixtures targeting genomes specific for a selected group of bacteria. Microflora DNA (mfDNA) was extracted from vaginal, oral and fecal clinical swabs. In addition forensic samples were processed. Vaginal samples showed a strong specific signal for bacteria of the female genital tract. Oral samples clearly showed signal for bacteria present in saliva, and in fecal samples the main signal was from Enterococcaceae. Vaginal casework samples showed results comparable to freshly collected ones; moreover the DNA extracted was successfully used for STR typing. Also mixtures of body fluids were analyzed, providing a microbiological signature compatible with the presence of microbes of oral, fecal and vaginal origin. The presented method can be useful in identifying biological fluids, and it is based on DNA technologies already available in forensic laboratories and feasible for further high throughput automation.

Swimming pools and fungi: an environmental epidemiology survey in Italian indoor swimming facilities.

A growing number of people attend swimming facilities for recreational activities, rehabilitative treatments, or sport. Filamentous fungi and yeast can be isolated from contaminated air, water and surfaces and may represent a biological risk for employees and users. Here we investigated the occurrence of mycotic species, in a sample of Italian swimming pools (n = 10). Detection and identification of isolated species were achieved by cultural and morphological methods. Results revealed moderate mycotic titres and a high biodiversity. Penicillium spp., Aspergillus spp., Cladosporium spp. and Alternaria sp., were constantly detected in air and surfaces sampled by the swimming area, while pathogenic yeast Candida albicans was never detected. Fusarium spp. was the most common taxon isolated from surfaces. For one facility, we typed the genotypic profiles and studied, by genetic typing, the spatial and temporal distribution of isolates. Phylogenetic relationships between species were analysed by alignment of small ribosomal subunit RNA sequences.