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Abstract The lower female reproductive tract is notoriously dominated by Lactobacillus species, among which Lactobacillus crispatus emerges for its protective and health‐promoting activities. Although previous comparative genome analyses highlighted genetic and phenotypic diversity within the L. crispatus species, most studies have focused on the presence/absence of accessory genes. Here, we investigated the variation at the single nucleotide level within protein‐encoding genes shared across a human‐derived L. crispatus strain selection, which includes 200 currently available human‐derived L. crispatus genomes as well as 41 chromosome sequences of such taxon that have been decoded in the framework of this study. Such data clearly pointed out the presence of intra‐species micro‐diversities that could have evolutionary significance contributing to phenotypical diversification by affecting protein domains. Specifically, two single nucleotide variations in the type II pullulanase gene sequence led to specific amino acid substitutions, possibly explaining the substantial differences in the growth performances and competition abilities observed in a multi‐strain bioreactor culture simulating the vaginal environment. Accordingly, L. crispatus strains display different growth performances, suggesting that the colonisation and stable persistence in the female reproductive tract between the members of this taxon is highly variable.

Abstract Although compositional variation in the gut microbiome during human development has been extensively investigated, strain-resolved dynamic changes remain to be fully uncovered. In the current study, shotgun metagenomic sequencing data of 12,415 fecal microbiomes from healthy individuals are employed for strain-level tracking of gut microbiota members to elucidate its evolving biodiversity across the human life span. This detailed longitudinal meta-analysis reveals host sex-related persistence of strains belonging to common, maternally-inherited species, such as Bifidobacterium bifidum and Bifidobacterium longum subsp. longum. Comparative genome analyses, coupled with experiments including intimate interaction between microbes and human intestinal cells, show that specific bacterial glycosyl hydrolases related to host-glycan metabolism may contribute to more efficient colonization in females compared to males. These findings point to an intriguing ancient sex-specific host-microbe coevolution driving the selective persistence in women of key microbial taxa that may be vertically passed on to the next generation.

Members of the genus Bifidobacterium are among the first microorganisms colonizing the human gut. Among these species, strains of Bifidobacterium breve are known to be commonly transmitted from mother to her newborn, while this species has also been linked with activities supporting human wellbeing. In the current study, an in silico approach, guided by ecology- and phylogenome-based analyses, was employed to identify a representative strain of B. breve to be exploited as a novel health-promoting candidate. The selected strain, i.e., B. breve PRL2012, was found to well represent the genetic content and functional genomic features of the B. breve taxon. We evaluated the ability of PRL2012 to survive in the gastrointestinal tract and to interact with other human gut commensal microbes. When co-cultivated with various human gut commensals, B. breve PRL2012 revealed an enhancement of its metabolic activity coupled with the activation of cellular defense mechanisms to apparently improve its survivability in a simulated ecosystem resembling the human microbiome.

Abstract In vitro gut cultivation models provide host-uncoupled, fast, and cost-efficient solutions to investigate the effects of intrinsic and extrinsic factors impacting on both composition and functionality of the intestinal microbial ecosystem. However, to ensure the maintenance and survival of gut microbial players and preserve their functions, these systems require close monitoring of several variables, including oxygen concentration, pH, and temperature, as well as the use of a culture medium satisfying the microbial nutritional requirements. In this context, in order to identify the macro- and micro-nutrients necessary for in vitro cultivation of the infant gut microbiota, a meta-analysis based on 1669 publicly available shotgun metagenomic samples corresponding to fecal samples of healthy, full-term infants aged from a few days to three years was performed to define the predominant species characterizing the “infant-like” gut microbial ecosystem. A subsequent comparison of growth performances was made using infant fecal samples that contained the most abundant bacterial taxa of the infant gut microbiota, when cultivated on 18 different culture media. This growth analysis was performed by means of flow cytometry-based bacterial cell enumeration and shallow shotgun sequencing, which allowed the formulation of an optimized growth medium, i.e., Infant Gut Super Medium (IGSM), which maintains and sustains the infant gut microbial biodiversity under in vitro growth conditions. Furthermore, this formulation was used to evaluate the in vitro effect of two drugs commonly used in pediatrics, i.e., acetaminophen and simethicone, on the taxonomic composition of the infant gut microbiota.

The frequency of locally transmitted dengue virus (DENV) infections has increased in Europe in recent years, facilitated by the invasive mosquito species Aedes albopictus, which is well established in a large area of Europe. In Italy, the first indigenous dengue outbreak was reported in August 2020 with 11 locally acquired cases in the Veneto region (northeast Italy), caused by a DENV-1 viral strain closely related to a previously described strain circulating in Singapore and China. In this study, we evaluated the vector competence of two Italian populations of Ae. albopictus compared to an Ae. aegypti lab colony. We performed experimental infections using a DENV-1 strain that is phylogenetically close to the strain responsible for the 2020 Italian autochthonous outbreak. Our results showed that local Ae. albopictus is susceptible to infection and is able to transmit the virus, confirming the relevant risk of possible outbreaks starting from an imported case.

Dengue (DENV) and Zika (ZIKV) viruses are mosquito-borne human pathogens. In Italy, the presence of the competent vector Aedes albopictus increases the risk of autochthonous transmission, and a national plan for arboviruses prevention, surveillance, and response (PNA 2020–2025) is in place. The results of laboratory diagnosis of both viruses by the National Reference Laboratory for arboviruses (NRLA) from November 2015 to November 2022 are presented. Samples from 655 suspected cases were tested by both molecular and serological assays. Virus and antibody kinetics, cross-reactivity, and diagnostic performance of IgM ELISA systems were analysed. Of 524 cases tested for DENV, 146 were classified as confirmed, 7 as probable, while 371 were excluded. Of 619 cases tested for ZIKV, 44 were classified as confirmed, while 492 were excluded. All cases were imported. Overall, 75.3% (110/146) of DENV and 50% (22/44) of ZIKV cases were confirmed through direct virus detection methods. High percentages of cross reactivity were observed between the two viruses. The median lag time from symptoms onset to sample collection was 7 days for both DENV molecular (range 0–20) and NS1 ELISA (range 0–48) tests, with high percentages of positivity also after 7 days (39% and 67%, respectively). For ZIKV, the median lag time was 5 days (range 0–22), with 16% positivity after 7 days. Diagnostic performance was assessed with negative predictive values ranging from 92% to 95% for the anti-DENV systems, and of 97% for the ZIKV one. Lower positive predictive values were seen in the tested population (DENV: 55% to 91%, ZIKV: 50%). DENV and ZIKV diagnosis by molecular test is the gold standard, but sample collection time is a limitation. Serological tests, including Plaque Reduction Neutralization Test, are thus necessary. Co-circulation and cross-reactivity between the two viruses increase diagnostic difficulty. Continuous evaluation of diagnostic strategies is essential to improve laboratory testing.

Intro: Dengue virus (DENV) is a mosquito-borne human pathogen endemic in 129 countries. In Italy, presence of the competent vector increases the risk of autochthonous transmission from imported cases, and a surveillance national plan for Arboviruses (PNA 2020-2025) is in place. The results of laboratory diagnosis of DENV in Italy from November 2015 to November 2021 are presented. Methods: Samples from 502 suspected DENV cases were tested by both molecular/antigen NS1 and serological assays, including Plaque Reduction Neutralization Test (PRNT). Samples were tested in parallel for DENV, Chikungunya (CHIKV)and Zika virus (ZIKV). Cases were classified based on laboratory results according to the Italian National Plan for Arboviruses (PNA) criteria. Findings: After evaluation of laboratory results, there were 140/502 confirmed and 7/502 probable cases (DENV cases). A positive molecular test was found in 27.1% of cases and 66.7% had a positive antigen NS1 test. The lag time from symptoms to collection was 4 and 6 days, respectively, with a higher range of day detection limit for antigen NS1. DENV cases gave 87.8% IgM and 82.9% PRNT positive/border line results: samples positivity of 72.20% and 33.9% in IgM and PRNT, respectively, was observed within the first 7 days of disease. Overall, 10.9% IgM positive results were not confirmed as positive in PRNT or molecular test among excluded cases (274/502). 26/502 suspected cases were both DENV and ZIKV positive by IgM and/or PRNT, making it difficult to provide a final differential diagnosis. None had a possible co-infection of DENV and CHIKV. Conclusion: DENV diagnosis by molecular/antigen tests is the gold standard but the sample collection time is a limitation. Serological tests are thus necessary. However, IgM results need PRNT confirmation. Co-circulation of DENV and ZIKV increases diagnostic difficulty. Continuous evaluation of diagnostic strategies is essential against future DENV autochthonous outbreaks.

Bifidobacteria are recognized as health-promoting bacteria that reside in the human gut, helping in the digestion of fiber, preventing infections, and producing essential compounds like vitamins. To date, Bifidobacterium animalis subsp. lactis, together with Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, and Bifidobacterium longum, represents one of the species that are used as probiotic bacteria. Despite the extensive and detailed scientific research conducted on this microbial taxon, the molecular mechanisms by which B. animalis subsp. lactis exerts health benefits to its host are still largely unknown. Thus, we dissected the genetic repertoire and phylogenetic relationship of 162 strains of B. animalis subsp. lactis to select a representative reference strain of this taxon suitable for investigating its interaction with the host. The B. animalis subsp. lactis PRL2013 strain, which was isolated by a mucosal sample of a healthy adult, was chosen as the reference of the monophyletic cluster of human origin and revealed a greater adhesion index than that observed for another B. animalis subsp. lactis strain used in the industry as a probiotic supplement. Transcriptomics analyses of PRL2013 strain, when exposed to human cell monolayers, revealed 291 significantly upregulated genes, among which were found genes predicted to encode extracellular structures that may directly interact with human cells, such as extracellular polymeric substances, wall teichoic acids, and pili. IMPORTANCE To date, many Bifidobacterium animalis subsp. lactis strains have been isolated from human fecal samples. However, their presence in these samples does not necessarily suggest an ability to colonize the human gut. Furthermore, probiotics of non-human origin may not effectively interact with the gut epithelium, resulting in transient bacteria of the gut microbiota. In vitro experiments with human cells revealed that B. animalis subsp. lactis PRL2013, an autochthonous member of the human gut, shows colonization capability, leading to future applications in functional foods. [ABSTRACT FROM AUTHOR]

Bifidobacteria are extensively exploited for the formulation of probiotic food supplements due to their claimed ability to exert health-beneficial effects upon their host. However, most commercialized probiotics are tested and selected for their safety features rather than for their effective abilities to interact with the host and/or other intestinal microbial players. In this study, we applied an ecological and phylogenomic-driven selection to identify novel B. longum subsp. longum strains with a presumed high fitness in the human gut. Such analyses allowed the identification of a prototype microorganism to investigate the genetic traits encompassed by the autochthonous bifidobacterial human gut communities. B. longum subsp. longum PRL2022 was selected due to its close genomic relationship with the calculated model representative of the adult human-gut associated B. longum subsp. longum taxon. The interactomic features of PRL2022 with the human host as well as with key representative intestinal microbial members were assayed using in vitro models, revealing how this bifidobacterial gut strain is able to establish extensive cross-talk with both the host and other microbial residents of the human intestine.

bARPA Lazio, 00187 Rome, ItalycARIANET, 20159 Milan, ItalydIndependent Scientific Consultant, 20126 MilanOdour nuisance impacting residents' life comfort is the result of the presence in air of numerous species, mainly gaseous, negatively interacting with the human olfactory system. It is well known that odour nuisance does not depend on the mean gas concentrations over long averaging times (of the order of one hour), but on their turbulent fluctuations, that can be some orders of magnitude greater than hourly averaged values. Currently, the direct measure of intensity and frequency concentration peaks can be carried out only for a few species and requires high-cost instruments, representing a serious challenge for Environmental Protection Agencies responsible for public health and environmental protection. This paper describes the intermediate-cost methodology developed by ARPA Lazio, the environmental agency responsible for the Italian administrative region which includes Rome, to monitor and assess the presence of odorous gases. It is based on mobile vehicles equipped with intermediate-cost analyzers for some of the main odorous gases, successfully deployed in more than fifteen field campaigns carried out all over the region in the last five years. As an example, results from one of the campaigns are also reported briefly. Such an effort is mainly motivated by the need to fulfil institutional duties, but it also supports the possibility of building a network of affordable instruments capable of routinely monitor odour nuisance impact over relatively large regions.

Poor air quality remains the largest environmental health risk in Europe, despite the EU policy efforts. Especially in cities, the synergistic interactions between the urban heat island and urban pollution result in premature mortality, associated with cardiovascular and respiratory diseases. Mediterranean urban areas are particularly susceptible under the consideration that the intensity, frequency, and duration of heat waves will increase due to climate change. The LIFE SIRIUS project designates that air quality management needs to go beyond traditional approaches in order to consider synergistic effects. This paper assesses the impact of temperature on daily mortality from 2004 to 2019 in the Republic of Cyprus with the use of a Generalized Additive Model (GAM). The association between mean daily temperature and mortality is nonlinear, implying that a prompt rise in deaths occurs when temperatures are high, while for colder temperatures, the effect is delayed. We report an inverted J-shaped relationship between mean temperature and mortality, with the most prominent effects on human health documented at low temperatures. The population under study appears to be acclimatized to local conditions, as mortality increases after 10 days of exposure to the environmental risk. The results of this study will assist in the definition of city-specific thresholds above which health warnings for the protection of the local population will be issued, in the framework of LIFE SIRIUS.

Abstract Integral Membrane Protein 2 B (ITM2B) is a type II ubiquitous transmembrane protein which role remains unclear. ITM2B mutations have been associated with different disorders: mutations leading to longer mutant proteins have been reported in two distinct Alzheimer-like autosomal dominant disorders with early-onset progressive dementia and cerebellar ataxia. Both disorders share neurological features including severe cerebral amyloid angiopathy, non-neuritic plaques, and fibrillary tangles as in Alzheimer disease. Our group reported a missense mutation in ITM2B, in an unusual retinal dystrophy with no dementia. This finding suggests a specific role of ITM2B in the retina. As the identification of retinal-specific ITM2B partners could bring new insights into the cellular functions of ITM2B, we performed quantitative proteomics of ITM2B interactome of the human retina. Overall, 457 ITM2B partners were identified with 8 of them involved in visual transduction. In addition, bulk Gene Ontology analyses showed that many ITM2B partners are involved in several other biological functions, such as microtubule organization, protein translation and interestingly, mitochondrial homeostasis. These data represent the first report of the ITM2B interactome in the human retina and may serve as a valuable inventory of new potential ITM2B partners for future investigations of ITM2B physiological functions and dysfunctions.

Knowledge of the precipitation contribution to the Antarctic surface mass balance is essential for defining the ice-sheet contribution to sea-level rise. Observations of precipitation are sparse over Antarctica, due to harsh environmental conditions. Precipitation during the summer months (November–December–January) on four expeditions, 2015–16, 2016–17, 2017–18 and 2018–19, in the Terra Nova Bay area, were monitored using a vertically pointing radar, disdrometer, snow gauge, radiosounding and an automatic weather station installed at the Italian Mario Zucchelli Station. The relationship between radar reflectivity and precipitation rate at the site can be estimated using these instruments jointly. The error in calculated precipitation is up to 40%, mostly dependent on reflectivity variability and disdrometer inability to define the real particle fall velocity. Mean derived summer precipitation is ~55 mm water equivalent but with a large variability. During collocated measurements in 2018–19, corrected snow gauge amounts agree with those derived from the relationship, within the estimated errors. European Centre for the Medium-Range Weather Forecasts (ECMWF) and the Antarctic Mesoscale Prediction System (AMPS) analysis and operational outputs are able to forecast the precipitation timing but do not adequately reproduce quantities during the most intense events, with overestimation for ECMWF and underestimation for AMPS.

ABSTRACT It has been widely reported that members of the genus Lactobacillus dominate the vaginal microbiota, which is represented by the most prevalent species Lactobacillus crispatus, Lactobacillus jensenii, Lactobacillus gasseri, and Lactobacillus iners. L. crispatus is furthermore considered an important microbial biomarker due to its professed beneficial implications on vaginal health. In order to identify molecular mechanisms responsible for health-promoting activities that are believed to be elicited by L. crispatus, we performed in silico investigations of the intraspecies biodiversity of vaginal microbiomes followed by in vitro experiments involving various L. crispatus strains along with other vaginal Lactobacillus species mentioned above. Specifically, we assessed their antibacterial activities against a variety of pathogenic microorganisms that are associated with vaginal infections. Moreover, coculture experiments of L. crispatus strains showing the most antibacterial activity against different pathogens revealed distinct ecological fitness and competitive properties with regard to other microbial colonizers. Interestingly, we observed that even phylogenetically closely related L. crispatus strains possess unique features in terms of their antimicrobial activities and associated competitive abilities, which suggests that they exert marked competition and evolutionary pressure within their specific environmental niche. IMPORTANCE The human vaginal microbiota includes all microorganisms that colonize the vaginal tract. In this context, a vaginal microbiota dominated by Lactobacillus and specifically by Lactobacillus crispatus is considered a hallmark of health. The role of L. crispatus in maintaining host health is linked to its modulatory activity toward other members of the vaginal ecosystem and toward the host. In this study, in vitro experiments followed by genetic analyses of the mechanisms used by L. crispatus in colonizing the vaginal ecological niche, particularly in the production of different antimicrobial compounds, were evaluated, highlighting some intriguing novel aspects concerning the genetic variability of this species. Our results indicate that this species has adapted to its niche and may still undergo adaptation to enhance its competitiveness for niche colonization.

Summary Dogs and cats have gained a special position in human society by becoming our principal companion animals. In this context, efforts to ensure their health and welfare have increased exponentially, with in recent times a growing interest in assessing the impact of the gut microbiota on canine and feline health. Recent technological advances have generated new tools to not only examine the intestinal microbial composition of dogs and cats, but also to scrutinize the genetic repertoire and associated metabolic functions of this microbial community. The application of high‐throughput sequencing techniques to canine and feline faecal samples revealed similarities in their bacterial composition, with Fusobacteria, Firmicutes and Bacteroidetes as the most prevalent and abundant phyla, followed by Proteobacteria and Actinobacteria. Although key bacterial members were consistently present in their gut microbiota, the taxonomic composition and the metabolic repertoire of the intestinal microbial population may be influenced by several factors, including diet, age and anthropogenic aspects, as well as intestinal dysbiosis. The current review aims to provide a comprehensive overview of the multitude of factors which play a role in the modulation of the canine and feline gut microbiota and that of their human owners with whom they share the same environment.

Abstract Background A recent systematic review identified very few studies on women’s views on how to improve the quality of maternal and newborn care (QMNC). This study aimed at exploring the suggestions provided by women, after hospital delivery in Italy, on how to improve the QMNC. Methods A questionnaire, containing open questions to capture suggestions on how to improve QMNC, was used to collect suggestions of mothers who gave birth a tertiary care referral hospital in Northeast Italy, between December 2016 and September 2018. Two authors independently used thematic analysis to analyse women’s comments, using the WHO Standards for improving the QMNC as framework for the analysis. Results Overall 392 mothers provided a total of 966 comments on how to improve the QMNC. Overall 45 (11.5%) women made suggestions pertinent to “provision of care”, 222 (56.6%) to the “experience of care”, 217 (55.4%) to “physical or to human resources”. The top five suggestions were: 1) increase presence of a companion during the whole hospitalization (28.3% of women); 2) improve bathrooms and showers (18.4%); 3) improve effective communication from staff (14.0%); 4) improve staff professionalism, empathy, and kindness (13.5%); 5) increase support and information on how to provide care to the newborn (11.2%). Overall, 158 (16.4%) suggestions could not be classified in any WHO Standards, and among these most (72.1%) were related to physical structures, such as: decrease the number of patients per room; create areas for visitors; avoid case mixing in the same room; reduce rooming-in/better support the mother. Overall 62 (15.8%) women expressed appreciations. Conclusions Collecting the women’s views on how to improve the QMNC after hospital delivery highlighted critical inputs on aspects of care that should be improved in the opinion of service-users. More investments should be made for establishing routine systems for monitoring patients experience of care. Data collected should be used to improve QMNC. WHO Standards may be further optimized by adding items emerging as relevant for women in high-income countries.