Milk Bioactive Compounds and Gut Microbiota Modulation: The Role of Whey Proteins and Milk Oligosaccharides.

A strong correlation between the occurrence of various pathological conditions and intestinal dysbiosis is supported by a range of strong evidence. Vice versa, many pathologies have been shown, in turn, to be responsible for alterations in the gut microbiota, a condition that can worsen illness outcomes and response to therapies. For these reasons, great efforts have been made, and studies are still ongoing, to elucidate the mechanisms underlying gut microbiota alterations and to search for pharmacologic or other strategies that can effectively restore the gut microbiota. In this narrative review, we examined the most significant literature on the role of some milk bioactive compounds, such as milk oligosaccharides and whey proteins, in modulating the composition of the gut microbiota and the underlying mechanisms of action, with the aim of investigating the impact of the microbiota changes mediated by these milk bioactive molecules on human health, and their potential use as therapeutics to treat or adjuvate the treatment of gut dysbiosis and associated pathologies.

Implication of Sodium Hypochlorite as a Sanitizer in Ready-to-Eat Salad Processing and Advantages of the Use of Alternative Rapid Bacterial Detection Methods.

The use of disinfection agents in the washing processing of ready-to-eat (RTE) vegetables, especially sodium hypochlorite, is a common industrial practice performed to enhance microbiological quality. However, some studies have reported a restart of bacterial growth and a substantial increase in bacterial load during early storage associated with the use of disinfection agents, which might represent a risk for consumers. We evaluated the effect of sodium hypochlorite on bacterial growth trends during the shelf-life in Lactuca sativa, simulating the industrial procedures for RTE vegetable packaging. Immediately after sodium hypochlorite treatment, an effective abatement of the bacterial load was observed, followed by a restart of growth throughout storage. After 5 days, the bacterial load was close to that reached by the control samples, indicating that the net increase in bacterial load was significantly higher in the treated samples. This might be ascribed to the reduction in competitive microflora and/or to the induction of adaptive responses by resting bacteria, which might select disinfectant-resistant bacteria. These findings elicit some concerns about the actual duration of the shelf-life; products might decrease their microbiological quality earlier during storage, pointing out the need to better clarify the impact of sodium hypochlorite as a sanitizer to closer consider its use in RTE vegetable processing. Furthermore, due to the importance of the rapid estimation of bacterial load and the early detection of foodborne pathogens throughout the food chain, the accuracy of the rapid bacteria detection method, the Micro Biological Survey (MBS), and its effectiveness for microbiological analyses of RTE vegetables were evaluated.

A new point-of-care test for the rapid antimicrobial susceptibility assessment of uropathogens.

Bacterial resistance to antimicrobials is considered a major issue worldwide. This condition may account for treatment failure of urinary tract infections, which are among the most common infections both in community and healthcare settings. Therapy against uropathogens is generally administered empirically, possibly leading to unsuccessful therapy, recurrence and development of antibiotic resistance. The reduction in analytical time to obtain antimicrobial susceptibility test (AST) results could play a key role in reducing the cost of healthcare, providing information about antibiotic efficacy and thus preventing from either exploiting new and expensive antibiotics unnecessarily or using obsolete and ineffective ones. A more rational choice among treatment options would hence lead to more effective treatment and faster resolution. In this paper we evaluated the performance of a new Point Of Care Test (POCT) for the rapid prediction of antimicrobial susceptibility in urine samples performed without the need of a laboratory or specialized technicians. 349 patients were enrolled in two open-label, monocentric, non-interventional clinical trials in partnership with an Emergency Medicine ward and the Day Hospital of two large healthcare facilities in Rome. Antibiogram was carried out on 97 patients. Results from analysis of urine samples with the POCT were compared with those from routine AST performed on culture-positive samples, displaying high accuracy (>90%) for all tested antimicrobial drugs and yielding reliable results in less than 12 hours from urine collection thus reducing analytical and management costs.

A narrative review of alternative transmission routes of COVID 19: what we know so far.

The Coronavirus disease 19 (COVID-19) pandemics, caused by severe acute respiratory syndrome coronaviruses, SARS-CoV-2, represent an unprecedented public health challenge. Beside person-to-person contagion via airborne droplets and aerosol, which is the main SARS-CoV-2's route of transmission, alternative modes, including transmission via fomites, food and food packaging, have been investigated for their potential impact on SARS-CoV-2 diffusion. In this context, several studies have demonstrated the persistence of SARS-CoV-2 RNA and, in some cases, of infectious particles on exposed fomites, food and water samples, confirming their possible role as sources of contamination and transmission. Indeed, fomite-to-human transmission has been demonstrated in a few cases where person-to-person transmission had been excluded. In addition, recent studies supported the possibility of acquiring COVID-19 through the fecal-oro route; the occurrence of COVID-19 gastrointestinal infections, in the absence of respiratory symptoms, also opens the intriguing possibility that these cases could be directly related to the ingestion of contaminated food and water. Overall, most of the studies considered these alternative routes of transmission of low epidemiological relevance; however, it should be considered that they could play an important role, or even be prevalent, in settings characterized by different environmental and socio-economic conditions. In this review, we discuss the most recent findings regarding SARS-CoV-2 alternative transmission routes, with the aim to disclose what is known about their impact on COVID-19 spread and to stimulate research in this field, which could potentially have a great impact, especially in low-resource contexts.

Coumarin-Induced Hepatotoxicity: A Narrative Review.

Coumarin is an effective treatment for primary lymphoedema, as well as lymphoedema related to breast cancer radiotherapy or surgery. However, its clinical use is limited in several countries due to the possible occurrence of hepatotoxicity, mainly in the form of mild to moderate transaminase elevation. It is worth noting that only a few cases of severe hepatotoxicity have been described in the literature, with no reported cases of liver failure. Data available on coumarin absorption, distribution, metabolism, and excretion have been reviewed, focusing on hepatotoxicity studies carried out in vitro and in vivo. Finally, safety and tolerability data from clinical trials have been thoroughly discussed. Based on these data, coumarin-induced hepatotoxicity is restricted to a small subset of patients, probably due to the activation in these individuals of alternative metabolic pathways involving specific CYP450s isoforms. The aim of this work is to stimulate research to clearly identify patients at risk of developing hepatotoxicity following coumarin treatment. Early identification of this subset of patients could open the possibility of more safely exploiting the therapeutical properties of coumarin, allowing patients suffering from lymphoedema to benefit from the anti-oedematous activity of the treatment.

Gal-3BP in Viral Infections: An Emerging Role in Severe Acute Respiratory Syndrome Coronavirus 2.

Galectin-3 binding protein (Gal-3BP) is a multifunctional glycoprotein involved in cell-cell and cell-matrix interactions known to be upregulated in cancer and various viral infections, including HIV-1, HCV, and SARS-CoV-2, with a key role in regulating the antiviral immune response. Studies have identified a direct correlation between circulating levels of Gal-3BP and the severity of disease and/or disease progression for some viral infections, including SARS-CoV-2, suggesting a role of Gal-3BP in these processes. Due to Gal-3BP's complex biology, the molecular mechanisms underlying its role in viral diseases have been only partially clarified. Gal-3BP induces the expression of interferons (IFNs) and proinflammatory cytokines, including interleukin-6 (IL-6), mainly interacting with galectin-3, targeting the TNF receptor-associated factors (TRAF-6 and TRAF-3) complex, thus having a putative role in the modulation of TGF-ß signaling. In addition, an antiviral activity of Gal-3BP has been ascribed to a direct interaction of the protein with virus components. In this review, we explored the role of Gal-3BP in viral infections and the relationship between Gal-3BP upregulation and disease severity and progression, mainly focusing on SARS-CoV-2. Augmented knowledge of Gal-3BP's role in virus infections can be useful to evaluate its possible use as a prognostic biomarker and as a putative target to block or attenuate severe disease.

Introduction to the Special Issue: Microbiological Safety and Quality of Foods.

Recent shifts in food production, processing and distribution, linked to the globalization of the food trade and the need to meet new consumers habits, are continuously challenging global food systems [...].

Antiviral properties of whey proteins and their activity against SARS-CoV-2 infection.

Native and chemically modified whey proteins and their peptide derivatives are encountering the interest of nutraceutical and pharmaceutical industries, due to the numerous properties, ranging from antimicrobial to immunological and antitumorigenic, that result in the possibility to employ milk and its protein components in a wide range of treatment and prevention strategies. Importantly, whey proteins were found to exert antiviral actions against different enveloped and non-enveloped viruses. Recently, the scientific community is focusing on these proteins, especially lactoferrin, since in vitro studies have demonstrated that they exert an important antiviral activity also against SARS-CoV-2. Up-to date, several studies are investigating the efficacy of lactoferrin and other whey proteins in vivo. Aim of this review is to shed light on the most relevant findings concerning the antiviral properties of whey proteins and their potential applications in human health, focussing on their application in prevention and treatment of SARS-CoV-2 infection.

Application of the Micro Biological Survey analytical method for the determination of bacterial load in cow raw milk.

The aim of this study was to evaluate the performance of "Micro Biological Survey - MBS Test" in the enumeration of bacterial load in cow raw milk. The MBS test is based on a colorimetric method recently developed and patented by "Roma Tre" University, Italy. The evaluation of the performance of the MBS method was carried out by comparison with plate count at 30°C (gold standard) and flow cytometry. Thirteen independent set of experiments were performed analyzing a total of 104 samples of cow raw milk with the selected methods. Results obtained using the MBS method are comparable with those obtained with the plate count method at 30°C (CFU/mL) and flow cytometry technology; in particular, the results obtained with the MBS method are very close to plate count's at 30°C. On the other hand, there are statistically significant differences between these two methods' and flow cytometry technology's results that could be due to the different experimental conditions.

Microbiological Quality of Ready-to-Eat Leafy Green Salads during Shelf-Life and Home-Refrigeration.

The market of ready-to-eat leafy green salads is experiencing a noticeable growth in Europe. Since they are intended to be consumed without additional treatments, these ready-to-eat products are associated with a high microbiological risk. The aim of this work was to evaluate the microbiological quality and safety of ready-to-eat leafy green salads sold in widespread supermarket chains in Lazio, Italy, on the packaging date during shelf-life and during home-refrigeration. The study also aimed to determine the differences between low-, medium-, and high-cost products. Salmonella spp. and L. monocytogenes were chosen as safety indicators as specified by European regulations while total aerobic mesophilic bacteria and Escherichia coli were chosen as quality indicators as suggested by national guidelines. Analyses were performed following the ISO standards and in parallel for the evaluation of total aerobic mesophilic bacteria, with an alternative colorimetric system, the Micro Biological Survey method, in order to propose a simple, affordable and accurate alternative for testing the microbiological quality of products, especially suitable for small and medium enterprises and on-site analyses. The study revealed high, unsatisfactory, total bacterial loads in all analyzed samples on the packaging date and expiry date and a very high prevalence of Salmonella spp. (67%) regardless of the selected varieties and cost categories; L. monocytogenes was not recovered aligning with the results obtained in other studies.

Rapid Microbiological Assessment in Raw Milk: Validation of a Rapid Alternative Method for the Assessment of Microbiological Quality in Raw Milk.

The consumption of dairy products and the dairy industry are one of the main global agri-food sectors for its size, economic importance, and level of technology. Microbiological quality of pasteurized milk or other milk products is dependent on microbiological quality of raw milk. A variety of microbiological count methods is available for monitoring the hygienic quality of raw milk. Among them, the pour plate method is the official essay for counting the number of colony-forming units in milk samples according to International Organization for Standardization (ISO) No. 4833-1:2013. The aim of the present study is the validation of the Micro Biological Survey (MBS) method, against the reference plate-count method, for the assessment of the microbiological quality of raw milk. This comparative study, performed in collaboration with the Istituto Zooprofilattico Sperimentale del Lazio e della Toscana M. Aleandri (IZSLT), demonstrates the accuracy of this alternative method for the determination of total viable bacterial count in cow's raw milk. The results obtained with the MBS method highlight its potential as a valid tool for reliable microbiological analysis in dairy industries.

A new point-of-care test for the rapid detection of urinary tract infections.

Urinary tract infections (UTIs) are among the most common infections in all age groups. Fast and accurate diagnosis is essential to ensure a timely and effective therapy. Alongside with reference culture-based methods, several point-of-care tests (POCTs) for early detection of UTIs have been developed, but they have not been significantly implemented in current clinical practice. The Micro Biological Survey (MBS) POCT is a simple test developed by MBS Diagnostics Ltd. (London, UK) for the detection and management of UTIs. The present study has been undertaken to investigate the potentials and limits of the MBS POCT. A total of 349 patients were enrolled in two open-label, monocentric, non-interventional clinical trials in collaboration with an Emergency Medicine department and the outpatient clinic of two hospitals in Rome. Results of urine analysis using the MBS POCT were compared with those of the routine culture-based tests for UTI diagnosis performed by the hospital laboratory. The MBS POCT provided fast results revealing high bacterial count UTIs (≥ 105 CFU/ml) with 97% accuracy, 92% sensitivity, 100% specificity, 99% PPV, and 96% NPV within a 5-h analytical time threshold.

Field application of the Micro Biological Survey method for the assessment of the microbiological safety of different water sources in Tanzania.

Access to safe water is stated within human rights as essential for life, as water can be a source of severe enteric infections threatening human health, in particular children from Developing Countries. Along with reference methods, need is pressing for alternative methods to flank reference ones to improve water safety on-site monitoring and in the absence of scientific facilities or even electricity supply. The Micro Biological Survey (MBS) method has already been successfully applied to water safety assessment in Developing Countries. A total of 18 water samples were collected from different sources (rivers, dug wells, tap water) within the Rukwa Region, Tanzania, and underwent analysis for Total Coliforms following the MBS method. Globally, rivers showed more frequently contamination, followed by dug wells, tap water and tanks. Results demonstrate the need for continuous monitoring of water sources, even in difficult frameworks lacking electric supply, to help improve control over water quality, possibly using alternative methods to simplify existing protocols.

Integrating running water monitoring tools with the Micro Biological Survey (MBS) method to improve water quality assessment.

Running water habitats are among the most altered aquatic systems by human activities driving an increase in the organic components and the associated bacterial load as well. To contribute in improving the monitoring activities in running waters, here we tested the validity of the new Micro Biological Survey (MBS) method to specifically assess the bacterial load in running waters focusing on Total Viable Counts (at 22°C and 37°C) and Escherichia coli (at 44°C) in order to propose a new prognostic tool for watercourses. MBS method is an alternative colorimetric method for counting bacterial load in water and food samples that is easy to use and leads to a reliable and simple interpretation of results, being also faster and less expensive than traditional methods. Then, we compared MBS with the traditionally used reference method for the bacterial load, and with the most used biotic index for Italian watercourses based on the benthic invertebrates: the Extended Biotic Index (EBI). The last comparison was performed to validate the use of MBS in biomonitoring activities since the benthic invertebrate multi-species assemblage (and then EBI) alter own structure mainly depending on the organic component variation. During the first part of the study, the assessment of both linearity (regressions among bacterial concentrations) and accuracy (significant correlation between a measured value and a value used as reference) confirmed the validity of the MBS method. Second, the linear regressions between the three investigated microbial parameters vs. both physical-chemical descriptors and EBI, revealed the usefulness of MBS as a valid tool for routine microbiological analyses involved in rapid and easy field monitoring activities. This represents the first attempt to evaluate the river microbial status by exploiting the innovative MBS on running waters to propose it as new valuable monitoring tool in the biomonitoring field.

Field Application of the Micro Biological Survey Method for the Assessment of the Microbiological Safety of Different Water Sources in Horn of Africa and the Evaluation of the Effectiveness of Moringa Oleifera in Drinking Water Purification.

Water monitoring requires expensive instrumentations and skilled technicians. In developing Countries as Africa, the severe economic restrictions and lack of technology make water safety monitoring approaches applied in developed Countries, still not sustainable. The need to develop new methods that are suitable, affordable, and sustainable in the African context is urgent. The simple, economic and rapid Micro Biological Survey (MBS) method does not require an equipped laboratory nor special instruments and skilled technicians, but it can be very useful for routine water analysis. The aim of this work was the application of the MBS method to evaluate the microbiological safety of different water sources and the effectiveness of different drinking water treatments in the Horn of Africa. The obtained results have proved that this method could be very helpful to monitor water safety before and after various purification treatments, with the aim to control waterborne diseases especially in developing Countries, whose population is the most exposed to these diseases. In addition, it has been proved that Moringa oleifera water treatment is ineffective in decreasing bacterial load of Eritrea water samples.

Field Application of the Micro Biological Survey Method for a Simple and Effective Assessment of the Microbiological Quality of Water Sources in Developing Countries.

According to the World Health Organization (WHO) guidelines, "safe drinking-water must not represent any significant risk to health over a lifetime of consumption, including different sensitivities that may occur between life stages". Traditional methods of water analysis are usually complex, time consuming and require an appropriately equipped laboratory, specialized personnel and expensive instrumentation. The aim of this work was to apply an alternative method, the Micro Biological Survey (MBS), to analyse for contaminants in drinking water. Preliminary experiments were carried out to demonstrate the linearity and accuracy of the MBS method and to verify the possibility of using the evaluation of total coliforms in 1 mL of water as a sufficient parameter to roughly though accurately determine water microbiological quality. The MBS method was then tested "on field" to assess the microbiological quality of water sources in the city of Douala (Cameroon, Central Africa). Analyses were performed on both dug and drilled wells in different periods of the year. Results confirm that the MBS method appears to be a valid and accurate method to evaluate the microbiological quality of many water sources and it can be of valuable aid in developing countries.

Water supply, sanitation and health risks in Douala 5 municipality, Cameroon.

BACKGROUND: In the poor zones of sub-Saharan Africa, the conventional drinking water network is very weak. The populations use alternative groundwater sources which are wells and springs. However, because of urbanization, the groundwater sources are degrading gradually making pure, safe, healthy and odourless drinking water a matter of deep concern. There are many pollutants in groundwater due to seepage of organic and inorganic pollutants, heavy metals as well as microbiological contamination. This study was carried out in October 2012 and aimed to analyze the practices and behaviors of populations related to water supply that may constitute potential risks of microbiological contamination and emergence of waterborne diseases. METHOD: This study was carried out on a sample of 285 households, distributed in twelve principal quarters of the Douala V municipality in Cameroon. After data collection through the questionnaires, the selection of vulnerable quarters was done by the tabulation of some specific results on the questionnaire. The microbiological analysis was carried out using an innovative rapid test called "Micro Biological Survey (MBS)" that has been developed and patented by MBS srl, a recent spin-off of the University of Roma Tre. RESULT: We found out that 42.30% of households used water from drilled wells (forages), 33.80% from Cameroon National Water Company (CDE) distribution network, 9.50% from spring, and 9.40% used other source of water such as buying industrial mineral water. However, it should be noted that, as dangerous as it may be, wells ("puits" in French) water is used as permanent source of drinking water by 5% of households. In general, 63.2% of households affirmed not to have benefited the fitting of public water point. Moreover, among the quarters were the households affirmed to have benefited from the development of a public water point, 52.5% of these water point were no longer functional. From the obtained data we have assessed that 83% of wells analyzed are faecally contaminated and all the forages analyzed (100%) are not contaminated. The most affected people by water-related diseases are children under 5 years (43%). CONCLUSION: We highly recommend the City Council of Douala V to restrict the use of the contaminated wells and provide alternative water sources such as forages for public use and to ensure that microbiological quality of the new and already existing water sources is controlled regularly by the Technical Service.

A new device for the prompt diagnosis of urinary tract infections.

BACKGROUND: Urinary tract infections (UTIs) are among the most common infectious diseases. RESULTS obtained from conventional microbiological analysis of urine and antibiotic susceptibility testing are available only after a few days, delaying precise diagnosis and appropriate therapy. Micro Biological Survey (MBS) srl (a spin-off of Roma Tre University, Rome, Italy) has developed and patented an automated colorimetric test for rapid bacterial counting. In a preliminary validation study it was demonstrated that the results obtained with the MBS method are equivalent to the results obtained with conventional culture-based microbiological analysis. METHODS: In this study, sterile urine samples were artificially contaminated with bacterial species that are most frequently responsible for UTIs. The MBS method was used to evaluate the presence of bacteria and their sensitivity to some of the most commonly used antibiotics in UTIs. RESULTS: The MBS method was able to detect in a few hours the presence or absence of bacteria at clinically significant concentrations (>105 CFU/mL), and to provide their susceptibility pattern to a limited panel of antibiotics. DISCUSSION: The results obtained demonstrate that the MBS point-of-care testing (POCT) device could be developed into a valuable aid for the management of UTIs, possibly addressing more precise diagnosis and appropriate therapy.

Microbiological safety and quality of Mozzarella cheese assessed by the microbiological survey method.

Dairy products are characterized by reduced shelf life because they are an excellent growth medium for a wide range of microorganisms. For this reason, it is important to monitor the microbiological quality of dairy products and, in particular, the total viable count and concentration of Escherichia coli, as they are indicators of the hygienic state of these products. In addition, in dairy products such as Mozzarella cheese, it is important to monitor the concentration of lactic acid bacteria (LAB), as they are the major components of starter cultures used in cheese production, contributing to the taste and texture of fermented products and inhibiting food spoilage bacteria by producing growth-inhibiting substances. For these reasons, to ensure the quality and safety of their products, cheese makers should monitor frequently, during fresh cheese production, the concentration of LAB and spoilage bacteria. However, usually, small- to medium-size dairy factories do not have an internal microbiological laboratory and external laboratories of analysis are often too expensive and require several days for the results. Compared with traditional methods, the microbiological survey (MBS) method developed by Roma Tre University (Rome, Italy) allows faster and less-expensive microbiological analyses to be conducted wherever they are necessary, without the need for a microbiological laboratory or any instrumentation other than MBS vials and a thermostat. In this paper, we report the primary validation of the MBS method to monitor LAB concentration in Mozzarella cheese and the analysis, using the MBS method, of total viable count, E. coli, and LAB concentrations in the production line of Mozzarella cheese as well as during the shelf life of the product stored at 20°C. The results obtained indicate that the MBS method may be successfully used by small- to medium-size dairy factories that do not have an internal microbiological laboratory. Using the MBS method, these dairy factories can monitor autonomously the microbiological safety and quality of their products, saving both time and money.