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The present study investigated the in situ microbial indicators' occurrence in water and biofilm in drinking water distribution systems of sport facilities such as gyms. The presence of Legionella, such as a potential pathogen, was also verified. Water and biofilm were contemporaneously collected and microbiologically analyzed. Few colonies of coliforms were detected in biofilm but not in the corresponding water samples. Conversely, some colonies of heterotrophs were counted at 22 degrees C in both the 80% biofilm samples and the 53% water samples. Legionella was detected in 29% of the hot water systems, and, in particular, L. pneumophila in 21% and L. non pneumophila in 9% of the samples. Higher counts were found in biofilm samples. As it was recommended for swimming pools, it is to be hoped that hygiene risk assessment criteria and safety plans could be also developed for gyms to reduce potential hazards to health for all attendants. [ABSTRACT FROM AUTHOR]

The spread of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in humans, animals and environment is a growing threat to public health. Wastewater treatment plants (WWTPs) are crucial in mitigating the risk of environmental contamination by effectively removing contaminants before discharge. However, the persistence of ARB and ARGs even after treatment is a challenge for the management of water system. To comprehensively assess antimicrobial resistance dynamics, we conducted a one-year monitoring study in three WWTPs in central Italy, both influents and effluents. We used seasonal sampling to analyze microbial communities by 16S rRNA, as well as to determine the prevalence and behaviour of major ARGs (sul1, tetA, bla TEM , bla OXA-48 , bla CTX-M-1 group , bla KPC ) and the class 1 Integron (int1). Predominant genera included in order: Arcobacter, Acinetobacter, Flavobacterium, Pseudarcobacter, Bacteroides, Aeromonas, Trichococcus, Cloacibacterium, Pseudomonas and Streptococcus. A higher diversity of bacterial communities was observed in the effluents compared to the influents. Within these communities, we also identified bacteria that may be associated with antibiotic resistance and pose a significant threat to human health. The mean concentrations (in gene copies per liter, gc/L) of ARGs and int1 in untreated wastewater (absolute abundance) were as follows: sul1 (4.1 × 10 9 ), tetA (5.2 × 10 8 ), bla TEM (1.1 × 10 8 ), bla OXA-48 (2.1 × 10 7 ), bla CTX-M-1 group (1.1 × 10 7 ), bla KPC (9.4 × 10 5 ), and int1 (5.5 × 10 9 ). The mean values in treated effluents showed reductions ranging from one to three log. However, after normalizing to the 16S rRNA gene (relative abundance), it was observed that in 37.5 % (42/112) of measurements, the relative abundance of ARGs increased in effluents compared to influents. Furthermore, correlations were identified between ARGs and bacterial genera including priority pathogens. This study improves our understanding of the dynamics of ARGs and provides insights to develop more effective strategies to reduce their spread, protecting public health and preserving the future efficacy of antibiotics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

In the present work, a simple two-step method is proposed for mixed oxide synthesis aimed at the achievement of antibacterial nanomaterials. In particular, Cu, Zn and Co have been selected to achieve single-, double- and triple-cation oxides. The synthesized samples are characterized by XRD, IR, SEM and EDX, indicating the formation of either crystalline or amorphous hydrocarbonate precursors. The oxides present one or two crystalline phases, depending on their composition; the triple-cation oxides form a solid solution of tenorite. Also, the morphology of the samples varies with the composition, yielding nanoparticles, filaments and hydrangea-like microaggregates. The antibacterial assays are conducted against E. coli and indicate an enhanced efficacy, especially displayed by the oxide containing 3% Co and 9% Zn incorporated into the CuO lattice. The oxides with the highest antibacterial properties are tested for their cytotoxicity, indicating a low toxicity impact, in line with literature data.

Several microorganisms can be found in tattoo inks injected into the skin, despite the ink matrix being considered inhospitable to microbial growth. Studies on the microbial quality of tattoo inks have reported the presence of microorganisms in most of the samples. This study aimed to assess the survival of environmental and human microbial species, selected on the specific criteria, in tattoo inks. Undiluted sterile black ink and serial dilutions (10-fold/100-fold) were each separately seeded with four bacterial strains (Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus pumilus, Mycobacterium fortuitum), one yeast (Candida albicans), and one mould (Fusarium solani). Their survival was periodically tested using cultural methods. No tested microorganisms were able to survive in undiluted ink, except for B. pumilus that survived up to 3 weeks. All the tested species, except for S. aureus, showed survivability for up to 10 weeks in 100-fold diluted inks, and P. aeruginosa, M. fortuitum, and C. albicans were even able to grow. B. pumilus and F. solani had good rates of survival even at the smallest dilution. The ability of microorganisms to survive and grow in tattoo inks could have health implications if contaminated ink dilutions are used during tattooing practices and stored for a long time., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

Cryptosporidium is a leading global cause of waterborne disease, with many reported outbreaks related to main water supplies. In August 2019, an outbreak of cryptosporidiosis involving 80 cases occurred among 114 vacationers in a small municipality located in the Tuscan-Emilian Apennines, north-eastern Italy. After excluding a potential food-borne outbreak, the epidemiological investigation focussed on the hypothesis of a waterborne outbreak. This was confirmed by the finding of Cryptosporidium oocysts in stools of the cases and in water samples from the municipal water network. Molecular characterisation revealed the zoonotic species Cryptosporidium parvum as the causative agent. A single subtype (IIdA25G1) was found among all cases, and in one of two positive water samples. The municipality's water supply used spring water that only received a disinfection treatment insufficient to inactivate the parasite. Possible entry means into the water mains were found through further environmental investigations. As these types of water supplies are particularly vulnerable to various environmental factors, a control system based on the risk assessment of each phase of the water supply chain is required to guarantee water safety. Effective methods for detection of protozoan pathogens, which are generally excluded from routine water supply analysis, should be applied.

There is increasing evidence that indoor air quality and contaminated surfaces provide an important potential source for transmission of pathogens in hospitals. Airborne hospital microorganisms are apparently harmless to healthy people. Nevertheless, healthcare settings are characterized by different environmental critical conditions and high infective risk, mainly due to the compromised immunologic conditions of the patients that make them more vulnerable to infections. Thus, spread, survival and persistence of microbial communities are important factors in hospital environments affecting health of inpatients as well as of medical and nursing staff. In this paper, airborne and aerosolized microorganisms and their presence in hospital environments are taken into consideration, and the factors that collectively contribute to defining the infection risk in these facilities are illustrated.

Careless use conditions of filter jugs were applied to simulate and evaluate the behavior of two ubiquitous aquatic bacterial species, Pseudomonas aeruginosa and Enterobacter aerogenes . According to a reference protocol, nine different jugs of popular brands sold in the Italian market were used for the test. Separately, a suspension of the two bacteria was spiked in water used for filling the jugs. The concentration of the test organisms and total aerobic microbial count (TAMC) was measured daily in the filtered water along a period corresponding to the cartridge lifetime. Results showed a different trend of bacterial behavior. E. aerogenes was detectable exclusively on the first day after jug filling, while P. aeruginosa confirmed its persistence over time in all the jugs and its ability to potentially colonize surfaces and cartridges. The TAMC was detected at a concentration range from 10 2 to 10 7 CFU/100 mL in all the tests, high values that were not far from those raised in bottled flat natural mineral water weeks after bottling.

Profiling bathing waters supported by Quantitative Microbial Risk Assessment (QMRA) is key to the WHO's recommendations for the 2020/2021 revision of the European Bathing Water Directive. We developed an area-specific QMRA model on four pathogens, using fecal indicator concentrations (E. coli, enterococci) for calculating pathogen loads. The predominance of illness was found to be attributable to Human Adenovirus, followed by Salmonella, Vibrio, and Norovirus. Overall, the cumulative illness risk showed a median of around 1 case/10000 exposures. The risk estimates were strongly influenced by the indicators that were used, suggesting the need for a more detailed investigation of the different sources of fecal contamination. Area-specific threshold values for fecal indicators were estimated on a risk-basis by modelling the cumulative risk against E. coli and enterococci concentrations. To improve bathing waters assessment, we suggest considering source apportionment, locally estimating of pathogen/indicator ratios, and calculating site-specific indicators thresholds based on risk assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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.

Background: Atypical or non-tuberculous mycobacteria (NTM) are considered opportunistic pathogenic bacteria. They are natural inhabitants of a broad variety of environmental reservoirs, including natural and treated waters. Due to the lack of Italian studies on these microorganisms in drinking water, an investigation was carried out on their occurrence and on species present in a municipal water distribution system in Rome. The presence of the traditional indicators of faecal contaminations and free living amoebae were also verified., Methods: Two different methodological approaches based on cultural and molecular techniques have been applied in parallel., Results: Mean concentrations of NTM obtained with the two analytical methodologies were roughly 102 CFU/L. Among the numerous NTM species identified, M. chelonae was the most frequently isolated. No correlation of NTM with indicators of faecal contamination and amoebae was found., Conclusions: Mycobacteria can be present even if routine tests show water to have an excellent hygienic quality.

Background: For the water analysis, for Pseudomonas aeruginosa a presumptive positive result can be achieved in 40- 48 hours using the traditional membrane filtration technique followed by an additional 24-48 hour confirmation stage. Conversely, the Pseudalert Quanti-Tray™ method can give confirmed results after 24-28 hours. In this case, actively growing strains of Pseudomonas aeruginosa show a confirmed positive result when a specific enzyme cleaving the substrate in the reagent produces a blue fluorescence under 365 nm ultraviolet light. A comparison of the performance of the Pseudalert respect to the standard method was conducted using statistical methods., Methods: Drinking water was analyzed in parallel with the membrane filtration technique using Pseudomonas CN agar (UNI EN ISO 16266) and the Pseudalert. Confirmation test are requested by the standard method and although Pseudalert Quanti-Tray™ gives confirmed results, all the positive isolates were also confirmed. Data were analyzed by statistical methods., Results: For drinking water, Pseudalert showed a very high sensitivity (98,8%) and a high percentage of specificity (96,8%). From a total of 889 positive isolates, a very high confirmation rates (99,3%) was calculated. Statistical analyses confirmed that the two methods were not statistically different., Conclusions: These results indicate that the Pseudalert produces confirmed results in a shorter time than the standard reference method allowing the detection of Pseudomonas aeruginosa with no further confirmation steps. It could be a valid alternative method for the water analysis.

Samples of soil, well water, and wastewater from a zootechnical farm, water after phytodepuration and maize plants (Zea mays) grown on soils irrigated with these different kinds of water were analysed for indicator bacteria and the protozoa Giardia and Cryptosporidium. Protozoa and bacterial indicators, except coliforms, were not recovered from well water samples. In the effluent from the zootechnical farm, high parasitological concentrations were observed, whilst water after the phytodepuration process showed a concentration reduction of two orders of magnitude. The high numbers of Giardia and Cryptosporidium in the zootechnical effluent could represent a potential risk for the spread of the pathogens. Nevertheless the environmental spread is minimized when data on soils and plants are observed. From the study, it emerges that this water treatment system could represent an alternative option to other conventional wastewater treatments and an economic and environmental advantage.

The Working Group "Movement Sciences for Health" of the Italian Society of Hygiene, Preventive Medicine and Public Health has promoted the Italian translation of the WHO Guidelines for Safe Recreational Water Environments, relating to coastal and freshwater environments. In this article the authors briefly summarize the main areas covered in the guidelines, namely drowning and injury, exposure to cold, heat and sunlight, water quality, contamination of beach sand, exposure to algae and their products, aesthetic issues, exposure to chemical and physical agents, dangerous aquatic organisms, monitoring and assessment hazards and risks.

Data on the occurrence of non-tuberculous mycobacteria (NTM), in parallel with those obtained for bacterial indicators and amoebae, are presented with the aim to collect information on the spread of NTM in drinking water distribution systems in Italy. Samples were collected from taps of hospitals and households in Central and Southern Italy. The concentration values obtained for the more traditional microbial parameters complied with the mandatory requirements for drinking water. Conversely, moderate-to-high microbial loads (till 300 CFU/L) were observed for the NTM. Positive samples were obtained from 62% of the investigated water samples. Analogous results were observed for amoebae showing a higher percentage of positive samples (76%). In terms of public health, the presence of mycobacteria in water distribution systems may represent a potential risk especially for vulnerable people such as children, the elderly or immunocompromised individuals.

Feedstock and compost samples were collected from twenty composting plants and analysed from the microbiological point of view. Faecal indicator organisms were determined in order to evaluate the efficacy of processes for the removal of pathogenic micro-organisms with similar survival characteristics and to verify their suitability as appropriate markers of microbial quality of composted products. In addition to the classical bacterial indicators, selected organisms, such as Salmonella, Giardia, Cryptosporidium, Clostridium spores and helminth ova, were investigated. Statistically significant differences in the removal of the different micro-organisms were observed with regard to both the different composting plants (P < 0.05) and feedstock composition (P < 0.05). In fact, compost obtained by feedstock containing sewage sludge was shown to have a better hygienic quality in comparison with compost containing green discards and municipal solid waste as raw matter. Giardia cysts, Cryptosporidium oocysts and helminth ova were not effective indicators of hygienic quality of compost, whereas Clostridium perfringens spores, because of their high resistance to treatments, could be considered as an additional model for assessing the composting process, especially with regard to more resistant pathogen reduction.

A comparative study on the efficacy of peracetic acid and sodium hypochlorite in inactivating resistant stages of microorganisms such as Cryptosporidium, Giardia and Cl. perfingens was carried out. Furthermore the evaluation of the potential reciprocal correlation among the concentrations of the organisms was performed. The results obtained indicate that, at the same experimental conditions, peracetic acid and sodium hypochlorite have nearly similar reduction power against the resistant stages of Giardia and Cl. perfringens. Both the oxidants are instead less efficient in the abatement of Cryptosporidium oocysts. Findings have also confirmed our previous studies on the absence of association between Clostridium and the protozoa.

Cryptosporidium oocysts and Giardia cysts has been isolated from waters worldwide. In Italy, studies on these parasites in the environment are still limited due to absence of epidemiological evidence and difficulty of adequate methodologies of sampling and analysis. The new Drinking Water Directive 98/83/CE states that Cryptosporidium has to be determined in water intended for human consumption if Clostridium perfringens is detected. This investigation contributes to the knowledge of both Cryptosporidium and Giardia occurrences in Italian aquatic environments through a two-year monitoring program. In addition, indicator bacteria and Clostridium perfringens were monitored in sewage, surface waters, drinking water, and swimming pools and possible correlations were calculated among all the selected parameters. Cysts and oocysts were detected in sewage and surface water and Giardia numbers always prevailed over Cryptosporidium. The parasites were not detected in drinking water, while a positive sample was obtained from the analysis of swimming pools. Pearson's correlation coefficients evidenced a reciprocal correlation between both the protozoa and the Enterococci counts.

Traditionally, the microbiological quality of waters has been measured by the analysis of indicator microorganisms. The article reviews the sanitary significance of traditional indicators of faecal contamination (total coliforms, faecal coliforms and faecal streptococci) and points out their limits. For some characteristics Escherichia coli may be considered a more useful indicator then faecal coliforms and recently it has been included in all recent laws regarding fresh, marine and drinking water. A clearer taxonomic definition of faecal streptococci evidenced the difficulty into defining a specific standard methodology of enumeration and suggested the more suitable role of enterococci as indicator microorganisms. Several current laws require the detection of enterococci. The resistance of Clostridium perfringens spores may mean that they would serve as a useful indicator of the sanitary quality of sea sediments.

Water samples were collected from a wastewater treatment plant and from two lakes in the vicinity of Rome (Italy) for detection and genotypic characterization of Giardia. All wastewater samples and all but two of the lake samples were scored by immunofluorescence as positive for Giardia. In addition, Giardia cysts were detected, by nested PCR, in all of the examined samples. Analysis of the SSU-rRNA gene sequences showed that all Giardia isolates belonged to Assemblage A. The presence of Assemblage A genotype in wastewater and surface bathing water represents an important public health concern. In fact, isolates showing this genotype are considered to be potential human pathogens and also have zoonotic implications.

In Italy controls on the hygienic quality of swimming pools are fixed by the Accordo tra il Ministero della Salute, le regioni e le province autonome di Trento e di Bolzano, come into force in 2003. In the present study swimming pools were investigated from the microbiological point of view on the basis of the new legislation. Contemporaneously, for the first time in Italy, analytical controls on the occurrence of the protozoa Cryptosporidium and Giardia were performed with both techniques of immunofluorescence and polymerase chain reaction. Results evidenced a good water quality when the bacterial parameters stated by law were considered. However in some swimming pools, the protozoa were also recovered. Factors affecting the use of molecular techniques on the analysis of environmental samples are also put into evidence.

An investigation on the hygienic quality of the Tiber river was conducted with the aim both to enumerate Giardia cysts and Cryptosporidium oocysts in the water and to determine possible correlations between them and bacterial indicators, pathogens and physico-chemical parameters. A low hygienic water quality was evidenced, with high counts of microorganisms. Furthermore, variable concentrations of Giardia and Cryptosporidium were observed. No correlation was found between the protozoa concentrations and that of the other microorganisms, whilst a significant correlation with redox potential and Giardia cysts was calculated.