Lactobacillus, Leuconostoc, Pediococcus, and Streptococcus were isolated, and the selection criteria encompassed their acidification capacity in fermentations carried out on chickpea, quinoa, and buckwheat flour extracts; the capacity to produce exopolysaccharides (EPS); and the antimicrobial activity against food spoilage molds and bacteria. Moreover, the stability of the LAB metabolites after the fermentation of the gluten-free flour extracts submitted to thermal and acidic treatments was also assessed.The occurrence of inflammatory responses in humans is frequently associated with food intolerances and is likely to give rise to irritable bowel disease. The use of conventional or unconventional flours to produce gluten-free baking doughs brings important technological and nutritional challenges, and the use of the sourdough biotechnology has the potential to overcome such limitations. In addition, the typical metabolic transformations carried out by Lactic Acid Bacteria (LAB) can become an important biotechnological process for the nutritional fortification and functionalization of sourdoughs due to the resulting postbiotics. In such a context, this research work aimed at isolating and selecting new LAB strains that resort to a wide range of natural environments and food matrices to be ultimately employed as starter cultures in gluten-free sourdough fermentations. Nineteen LAB strains belonging to the genera of  There is an increasing interest of consumers for emerging strategies that are able to promote a healthy lifestyle and the healing of certain diseases through non-invasive methods. In such a circumstance, sourdough bread and other sourdough-based baking goods , are enjoying an increasing popularity among consumers as convenient, nutritious, low-processed, natural, and healthy food.The consumption of functional foods with therapeutic and health-promoting effects support the new consumer demands. In general, consumers are already acquainted with the concepts of prebiotics and probiotics due to their intensive promotion by the pharmaceutical, nutraceutical, and agri-food industries. Simultaneously, healthy approaches are needed to overcome the massive antibiotic drugs administration and their side-effects on the colon’s microbiota of mammals .Probiotics are defined as living microorganisms able to exert beneficial effect on consumers’ health. Thus, probiotics should be constantly included in the diet in order to observe health improvements after long-term consumption. Prebiotics, on the other hand, known also as dietary fibers are carbohydrate-based substrates selectively fermented by probiotics in the colon ,3,4.New concepts of paraprobiotics and postbiotics are intensively studied, paraprobiotics being defined as killed or “ghost” probiotics and postbiotics being any water-soluble compound biosynthesized by probiotics, including Lactic Acid Bacteria (LAB) strains, during fermentations, or any other type of by-products from the bacterial cells after their lysis by physical, chemical, and/or enzymatic agents that possess health-promoting effects to the host ,7,8,9,10The beneficial effects of prebiotics, probiotics, postbiotics, and paraprobiotics could be maximized in complex functional foods, that include all the above-mentioned bioactive compounds, consumers being interested in this kind of products ,12.Furthermore, there is a correlation between the gastro-intestinal microbiota and a healthy lifestyle due to the fact that any perturbation on the typical gastro-intestinal microbiome may lead to unwanted immune responses that, ultimately, may degenerate into health imbalances and diseases ,13,14. DPeople suffering from irritable bowel diseases must include in their diets only gluten-free foods ,17. MoreGluten is a proteinaceous material, which can be separated from flour when the starch and other minor compounds are removed by washing with water ,28. It cPseudocereals and legumes are valuable sources of proteins, fats, and dietary fibers which can be used for novel gluten-free food products formulation. Their functional properties can be improved by lactic acid fermentation, a process which also enhances the nutritive and sensorial characteristics of the final fermented products. It can be used simply to decrease the risk of cross-contaminations with traces of gluten in food products. As previously mentioned, sourdough LAB is a source of proteolytic enzymes activated by acid production and, subsequently, is likely to be able to eliminate gluten toxicity during breadmaking. In fact, some lactobacilli possess a high potential to degrade Pro- and Gln-rich gliadin oligopeptides in wheat, which are responsible for the human inflammatory responses as a reaction to gluten ingestion ,26. MoreUnlike wheat doughs, where addition of sourdough is optional, the absence of gluten in rye flour makes the addition of sourdough a prerequisite to achieve good baking features. Sourdough fermentation is associated with the development of desired tastes and aroma and extension of shelf-life, as well as numerous nutritional and technological improvements during fermentation and baking. Traditional processes for sourdough fermentation are based on the continuous propagation (or back-slopping) of a fermented dough  from batch to batch, which enables a good chemical, functional, and microbiological stability and metabolic activity of the sourdoughs, thus contributing to a constant bread quality. Nevertheless, these processes are typically laborious, time-consuming and frequently poorly reproducible, making single or mixed (or co-cultured) microbial starter cultures an attractive alternative for bakeries ,26,39,40Nowadays, researchers are often particularly focusing their attention on the discovery and characterization of new LAB strains able to biosynthesize postbiotics such as exopolysaccharides (EPS), bioactive peptides, and short-chain fatty acids, in order to exploit their antimicrobial and functional properties in foods ,43,44,45Raw materials for the prospection and isolation of LAB, depicted in The LAB strains were isolated from various raw-materials  followinwsample/vwater). The ingredients were mixed with a sterile spatula and the resulting homogenous baking dough let to stand and ferment under controlled conditions in an incubator at 37 °C for 24 h. The resulting sourdough was propagated (backslopped) one time by mixing with a mixture of the same flour and water in the proportion of 20 g sourdough to 40 g of flour and 40 mL of tap water.In the case of the raw samples of some cereal, legume, and pseudocereal flours see , e.g., ww/v) NaCl saline solution.Depending on the used protocol, the fermented broth media and decimal dilutions from sourdoughs were used for further inoculation, growth, isolation, and purification of LAB. Decimal dilutions were obtained using a sterile 0.9% (w/v) CaCO3 as well as with 150 mg/L of cycloheximide, so as to prevent the growth of yeasts and molds. Inoculated MRS agar Petri dishes were further incubated at 37 °C for 48–72 h under aerobiosis.The inoculation of each of the previous samples was carried out with an inoculation loop by the streak-plate technique into Petri dishes of MRS agar supplemented with 1% , were selected and streaked in MRS agar and incubated under the same conditions in order to obtain pure cultures. When necessary, the isolates were properly purified by sub-culturing on MRS agar. Microscopic observation of fresh cells and Gram staining of the cells was carried out to verify the purity of the microbial isolates.Some morphological and biochemical assays were performed in order to establish the taxonomy of the isolated strains. Thus, the morphological examination was performed with a phase contrast microscope . The Gram staining was done following the standard procedure described by Collins and Lyne (2004)  using a 2O2 was poured. The effervescence of the oxygen on the slides was considered positive. For the urease test, the culture medium proposed by Foroudandeh et al. (2010) [Lactobacillus spp. The Gram-positive, catalase- and urease-negative cocci were preliminarily classified as Leuconostoc spp. [Streptococcus spp., and the spherical cocci associated in tetrades were preliminarily classified as Pediococcus spp. [For the catalase test, the protocol used by Pyar and Peh (2013)  was foll. (2010)  was usedtoc spp. ,57,58. Gcus spp. ,60.v/v) glycerol solution until further use [v/v) fresh inoculum containing approximately 109 CFU/mL (OD600nm = 1.8) was used in the subsequent metabolic assays according to the method reported by Lepczyńska and Dzika (2019) [The pure LAB cultures were grown under sterile conditions in MRS broth at 37 °C for 72 h, and stored at −80 °C in an ultra-freezer  in sterile capped plastic tubes  with 40%  , slightlThe indicator microorganisms (molds and bacteria) for the antimicrobial activity assay were isolated from different foods and raw sources, following similar procedures as those described elsewhere ,64,65,66Penicillium spp. strain was isolated from moldy wheat bread as follows: the mold spores were collected from the surface of the bread and streaked on Potato Glucose Agar according to the procedures described previously [Aspergillus niger and Aspergillus flavus strains were isolated from quinoa microbiota by spreading some seeds on Potato Glucose Agar dishes [w/v) NaCl saline solution by stirring with a vortex mixer . The spore’s suspension was then mixed at a proportion of 1:1 (v/v) with a 40% (v/v) glycerol solution in sterile capped plastic tubes and stored at −80 °C until further use.A eviously ,67. Asper dishes  and furtBacillus spp. strain was isolated from the quinoa seeds microbiota following the procedures described before [w/v) NaCl saline solution, stirred in vortex mixer and subjected to heat treatment of 80 °C for 10 min on a water bath to destroy any vegetative forms and keep only the spores. After cooling, 1 mL of the previous solution was inoculated by spread and streak plate techniques on Petri dishes of Plate Count Agar. After an incubation at 37 °C for 48 h, pure cultures were obtained by successive sub-culturing using the streak plate technique on the same culture medium, and the purity of the bacterial culture was checked with a phase contrast microscope. The pure culture of the indicator bacteria was grown under sterile conditions on Plate Count Agar slants at 37 °C for 48 h in aerobiosis conditions. Gram-positive rods, catalase- and urease-negative, were selected and preliminarily classified as Bacillus spp. [v/v) glycerol solution in a ratio of 1:1 (v/v) and stored at −80 °C in sterile capped plastic tubes  until further use.Finally, the endospore-forming Gram-positive rods d before . Brieflylus spp. . For theThe capacity of the newly and pure LAB strains to produce exopolysaccharides was evaluated using the methodologies described by Bachtarzi et al. (2019)  and ZhaovLAB/vmedium) and incubated at 37 °C for 72 h under aerobic conditions. Afterwards, the bacterial cells were discharged by centrifugation  at 6000 rpm and 4 ˚C for 10 min, and the content of extracellular exopolysaccharides was determined in the collected supernatant by the phenol-sulfuric acid assay using glucose as a standard, as described elsewhere [w/v) phenol solution in distilled water. The mixture was thoroughly vortexed and then 5 mL of 96% (v/v) H2SO4 aqueous solution was added. After mixing in the vortex mixer, all the reagents and samples were set aside at room temperature for 30 min. The absorbances of the samples were determined spectrophotometrically at the visible wavelength of 490 nm and taking into consideration the dilution factor of 100. The results were expressed as milligram of total sugars per milliliter of medium.Moreover, a volume of 0.18 μL of each selected LAB strain previously reactivated in MRS broth was then aseptically inoculated in sterile glass tubes containing 9 mL of modified MRS broth  were mixed with distilled water on a rotary shaker  at 25 °C, 100 rpm for 30 min. Then the liquid (supernatant) fraction was filtered under vacuum using Whatman no. 1 filter paper  and then sterilized  at 121 °C for 15 min. The pure LAB strains were reactivated, under aerobiosis, in MRS broth at 37 °C for 48 h, as previously described.The selected pure strains of the new LAB were assessed for their ability to produce organic acids in sterile broths extracted from the flours of the pseudocereals quinoa and buckwheat and the vegetable chickpea, following the procedures described previously  with slivLAB/vextract). Throughout the 72 h of aerobic fermentation at 37 °C, the total titratable acidity (TTA) was determined with an automatic titrator , at the beginning of the flour extract fermentation (t0) and every 24 h for 3 days. Furthermore, for the TTA determination, a volume of 10 mL of the fermented sample was mixed with 90 mL of distilled water, and the results were expressed as the volume of NaOH 0.1N (mL) required to reach the pH value of 8.5 [For the fermentation assays, 150 mL of sterile flour extract broths in a 300 mL Erlenmeyer were inoculated, under aseptic conditions, with 3 mL of the previous fresh pure LAB strains, giving rise to the ratio of 2%  solution supplemented with 0.1% (v/v) Tween 80, according to the slightly modified methodologies of Cizeikiene et al. (2013) [5 spores/mL, counted using a Thoma counting chamber. After the solidification of the media, several wells were created in the solid medium, and 50 µL of the supernatant (without cells) of each fermented flour extract, obtained after centrifugation at 6000 rpm for 10 min at 4 °C (see The strains of . (2013)  and Garc. (2013) . In each4 °C see , was aseBacillus spp., used as indicator, was reactivated in Nutrient Broth  at 37 °C overnight. For the agar well diffusion assays, a volume of 100 µL of the previous overnight inoculum (OD560 nm = 0.08) was used for inoculation into Petri dishes with Plate Count Agar through the surface spread plate technique. Afterwards, wells were created and 50 µL of the supernatant of each fermented flour extract  CFS, treated as mentioned above (see 5 spores/mL were spotted into the centers of the dishes. The CFS from the sterile flour extracts (unfermented) were used for the control samples. After the incubation of the Petri dishes for 96 h at 25 °C, the inhibition ratio was calculated using Equation (1):C is the diameter (in mm) of the mycelial growth in the control sample, and AT is the diameter (in mm) of the mycelial growth in the medium supplemented with the treated sample [The indicator strains of ibed see . The melbove see  and thend sample ,82.Bacillus spp. following the methodologies previously described (The CFS subjected to different temperatures and acidification levels  were useescribed .n = 3). Furthermore, regarding the statistical treatment, a Principal Component Analysis (PCA) of the experimental data was performed using Minitab 17  with the following variables: (1) Antifungal activity of the chickpea flour extract against Aspergillus niger, (2) Antifungal activity of the chickpea flour extract against Aspergillus flavus, (3) Antifungal activity of the chickpea flour extract against Penicillium spp., (4) Antibacterial activity of the chickpea flour extract against Bacillus spp., (5) Acidification capacity measured as TTA after a 72 h fermentation on the chickpea flour extract, (6) Antifungal activity of the quinoa flour extract against Aspergillus niger, (7) Antifungal activity of the quinoa flour extract against Aspergillus flavus, (8) Antifungal activity of the quinoa flour extract against Penicillium spp., (9) Antibacterial activity of the quinoa flour extract against Bacillus spp., (10) Acidification capacity measured as TTA after a 72 h fermentation on the quinoa flour extract, (11) Antifungal activity of the buckwheat flour extract against Aspergillus niger, (12) Antifungal activity of the buckwheat flour extract against Penicillium spp., (13) Antibacterial activity of the buckwheat flour extract against Bacillus spp., (14) Acidification capacity measured as TTA after a 72 h fermentation on the buckwheat flour extract, (15) Production of exopolysaccharides.The experiments were carried out in triplicate, and the results were expressed as mean value ± standard deviation (SD) for three independent measures  in any of the tested fermentation substrates . Moreover, all the fermented extracts with the strains no. 1, 2, 30, 33, 36, and 43 displayed a very good antibacterial activity against the endospore forming Gram-positive rods Bacillus spp. The buckwheat and quinoa flour extracts fermented with the strains no. 29, 44, and 48 possessed the best antibacterial activity.The LAB strain no. 24 revealed a very good antibacterial activity against The main aim of the current research work was to unfold the most favorable LAB strains to be further employed as single or co-cultured (multi-strain) starters in the gluten-free sourdough production. To that purpose, a set of fermentation variables (15) were included in the PCA, as the LAB acidification capacity in the fermentations carried out on chickpea, quinoa, and buckwheat flour extracts, the LAB capacity to produce exopolysaccharides and the antimicrobial  activity of the resulting fermented flour extracts. This study highlighted the distribution of the analyzed variables and lactic acid bacteria strains depicted in Lactobacillus spp. produced the highest amounts of organic acids that influenced the total titrable acidity (TTA) when chickpea, quinoa, and buckwheat flour extracts were fermented by these presumptive lactic acid bacteria strains . Strain no. 30 could be considered a good candidate due to its antimicrobials d when thLeuconostoc spp. was selected for its multiple capacities to acidify the chickpea and buckwheat flour extracts, for the exopolysaccharides production, and finally for the antimicrobial properties, all these features being observed on PC 1 , whereas a higher inhibition ratio (I = 12.12%) against the same mold was calculated for the chickpea flour extract fermented by strain no. 48. Strain no. 24 also showed an antagonistic effect against Aspergillus niger, the highest inhibition ratio (I = 20.19%) being calculated for the fermented chickpea flour extract that had a pH = 5.5.The stability of the antimicrobial compounds after thermal and acidification treatments of the cell-free extracts was assessed by fermentation with two selected LAB strains, specifically with the eatments  was diffAspergillus niger in the absence of additional treatments in the fermented quinoa flour extract. Strain no. 48 demonstrated an inhibition ratio of 6.93% against Penicillium spp. when the same extract was used. Furthermore, both strains showed a significant inhibitory activity against Penicillium spp. when the fermented quinoa extract was treated at 60 °C for 15 min, the highest inhibition ratio (I = 17.33%) belonging to strain no. 24.LAB strain no. 24 demonstrated a weak inhibitory activity (I = 0.82%) against Aspergillus niger was inhibited by the fermented extract obtained with LAB strain no. 24 with a ratio of 7.20%, here concluding that the antifungal activity could be related to other biosynthesized compounds that are stable under those conditions, except the organic acids.Moreover, antifungal activity was detected for strain no. 24 that was used for the fermentation of the quinoa flour extract that was subjected afterwards to sterilization . In this case, Aspergillus niger. Furthermore, in the fermented buckwheat broth heated at 80 °C for 15 min, various heat-stable compounds are also present with an inhibitory effect against the Penicillium spp., with inhibition ratios of 9.88% and 29.44%, respectively.After the fermentation of the buckwheat flour extract for 72 h at 37 °C by both of the selected LAB strains that were tested, i.e., strains no. 24 and 48, respectively, it can be concluded that within the fermented broth, several antifungal compounds exist, molecules that were stable at pH 7.5 with inhibition ratios of 7.02% and 10.53% against Bacillus spp. on the fermented buckwheat flour extract. The diameters of the inhibition zones were greater than 20 mm for all the flour extracts fermented by strains no. 24 and no. 48 at 60 °C for 15 min. Therefore, the antibacterial compounds that remained active after the thermal treatment at 121 °C or at the pH values of 3.5, 5.5, and 7.5 were not produced by the lactic acid bacteria strain no. 48 on the fermented buckwheat flour extract.The antibacterial activity of the samples fermented by LAB strain no. 24, without any other treatment, was observed only for the fermented quinoa flour extract , whereasPenicillium spp. of the untreated chickpea flour extract fermented with the lactic acid bacteria strain no. 48. This strain inhibited neither the Bacillus spp., nor the fungal strains included in the study of the untreated (only fermented) buckwheat flour extract. Even though the highest percentage of inhibition was calculated for the antifungal activity against Penicillium spp. and Aspergillus niger by the buckwheat flour extract fermented by strain no. 48 and then subjected to different treatments (80 °C or pH 3.5), the antibacterial activity displayed no positive results. Hence, the Penicillium spp. development was inhibited at a percentage of 25.60% when the temperature of 80 °C was applied for 15 min on the quinoa flour extract fermented by LAB strain no. 24. In addition, the sample mentioned above determined a very good inhibition of the Bacillus spp. growth under the same conditions.Taking into account the overall antimicrobial activity, it can be observed that the highest antibacterial activity of the untreated samples (16.67 mm zone of inhibition) was correlated to the highest inhibition ratio (I = 12.12%) against the Pediococcus spp. and Lactobacillus spp. These microorganisms proved to be able to inhibit the Aspergillus spp. growth and, therefore, to avoid the production of their mycotoxins. Other lactic acid bacteria strains with antimicrobial effects against several pathogens were found, for instance, in some spontaneous wheat, rye, and sorghum sourdoughs [Lactobacillus spp., followed by Pediococcus spp. and Leuconostoc spp.—their prevalence was thought to be influenced by the origin of the sourdough [The nineteen wild presumptive LAB strains were isolated from various sources such as fermented products, seeds, cereals, flours, and soil . By explurdoughs ,86,87,88urdoughs ,90. Furtourdough . Other aourdough . In thisourdough ,61,93, lTotal titratable acidity is a key parameter in the sourdough fermentation because it reveals the extension of the organic acids and other valuable antimicrobial compounds production, which, in turn, plays important roles in the sourdough and bread industry, such as antimicrobial and antioxidant properties which contribute to the final taste and aroma, increase the shelf-life, etc. The values of TTA observed in this work  were notLactobacillus spp. no. 2, 24, and 30 and the Leuconostoc spp. no. 48 due to their superior properties regarding acidification, antimicrobial properties, and exopolysaccharides production.The PCA with all the 15 variables  was usedStreptococcus termophillus lactic acid bacteria strain to produce EPS  compared to the ropy Lactococcus lactis lactic acid bacteria strain, in particular, 404 against 354 mg of glucose equivalents per liter.The screening among all the isolates of the presumptive lactic acid bacteria strains presented in Lactobacillus plantarum strain originating from cassava peel heaps produced more than 10 g/L exopolysaccharides. In the present study, the largest LAB producers, viz., strains no. 36 (Lactobacillus spp.) and no. 48 (Leuconostoc spp.), yielded more than double of those amounts.In this research work, the results of the phenol-sulfuric acid assay were used to detect and select the highly EPS-productive strains, even though the ropy visual (and to the touch) appearance is the most commonly used method to select the strains, being usually followed by the precipitation of the EPS and the determination of its sugar content by the phenol-sulfuric acid assay. In a study conducted by Amao et al. (2019) , a LactoLeuconostoc strains with interesting capacities to produce exopolysaccharides. In the studies carried out with human breast milk [Enterococcus faecalis and Lactobacillus spp. strains yielded amounts of EPS ranging from 380 to 737.3 mg/L. In the studies by Adebayo-Tayo et al. (2018) [Lactobacillus delbrueckii has been shown to be more effective (5.57 g/L). Nonetheless, in the same study [Weissella confusa were obtained.Furthermore, fermented meat, fruits, and dairy products were demonstrated to be good sources for the isolation of ast milk ,115,116,. (2018)  and Adeb. (2018) , the EPSme study  the EPS Lactobacillus paracasei, having obtained a maximum yield of 932 mg/L. Moreover, the carbon-to-nitrogen ratio among many other growth parameters proved to have a major impact on the amounts and quality of the EPS [The EPS production yields by the presumptive LAB in the current work were frequently higher than those described in the literature, the maximum value (approximately 22 g/L) being attained by strains no. 36 and 48. For future studies, the EPS will be characterized in terms of the type  as well as in terms of their structural and functional properties and the fermentation conditions optimized to yield high rates of EPS. Li et al. (2020)  evaluate the EPS .Lactobacillus plantarum and Lactobacillus delbrueckii ssp. bulgaricus, cultivated on wheat dough soluble extracts, and found out that the Lactobacillus plantarum strain had a better inhibitory effect against Penicillium spp., whereas a weaker inhibition (+) was observed against Aspergillus niger. The LAB strains included in this study [Aspergillus niger, Penicillium spp., and Aspergillus flavus, used as the indicator fungal strains.Luz et al. (2019)  studied is study  were furLeuconostoc mesenteroides, Lactobacillus curvatus, and Lactobacillus brevis showed the absence of an inhibitory effect against Aspergillus fischeri and a weak inhibition against Penicillium spp. On the other hand, in the study by Sun et al. (2020) [Lactobacillus spp. displayed good inhibitory capacity against Aspergillus niger, although most of the tested strains showed a better inhibition against Penicillium citrinum, Aspergillus flavus, Aspergillus fumigatus, and Fusarium graminearum. According to the study reported by Omedi et al. (2019) [Aspergillus niger and Penicillium chrysogenum. Nonetheless, the same work [Lactobacillus pentosus strains could not inhibit Aspergillus niger, in contrast to some strains of Pediococcus pentosaceus, where reasonable inhibition effects were observed.In a study by Bartkiene et al. (2019) , lactic . (2020) , three L. (2019) , the micame work  demonstrWeissella cibaria could inhibit the growth of Bacillus subtilis.In a study led by Dentice Maidana et al. (2020) , only twBacillus spp. The results are in accordance with the studies performed by Kaya and Simsek (2019) [Lactobacillus against Bacillus cereus. In contrast, in the study conducted by Verón et al. (2017) [Lactobacillus strains could not inhibit the growth of the spoilage bacterial strain Bacillus spp.In the present work, the tested lactic acid bacteria strains showed a very satisfactory inhibition capacity against k (2019)  and Salok (2019)  who repo. (2017) , most ofLactobacillus spp. (strain no. 24) and Leuconostoc spp. (strain no. 48) strains for 72 h at 37˚C and the CFS subjected to the abovementioned treatments to ascertain the stability of the antimicrobial compounds on such complex food matrixes.Another important objective in the present work was to evaluate the antimicrobial properties of the chickpea, quinoa, and buckwheat fermented flour extracts without cells  after being submitted to different levels of thermal  and acidic  treatments. The aerobic fermentations of the sterile flour extracts were undertaken with the Aspergillus niger, not only at pH values of 3.5 (I = 13.16%) but also at the pH values of 7.5 (I = 10.53%). A similar behavior of another lactic acid bacteria strain was reported by Varsha et al. (2014) [Aspergillus niger (I = 7.20%) was observed. Nevertheless, such antimicrobial compounds represent important technological features to the baking industry. Conversely, the fermented chickpea flour extract by lactic acid bacteria strain no. 48 unfolded an inhibitory ratio of 12.1% and 11.1% against Penicillium spp. in the absence of the treatments or when heated at 80 ˚C for 15 min, respectively. In addition, the quinoa flour extract fermented by lactic acid bacteria strain no. 24 that was subjected to a thermal treatment at 80 °C for 15 min exposed an inhibitory ratio above 25% against Penicillium spp.The results demonstrate that the buckwheat flour extract fermented with lactic acid bacteria strain no. 48 showed a great antifungal activity against . (2014) , who demAs already mentioned, these preliminary assays on the stability of antimicrobial compounds may suggest the presence of lactic acid bacteria metabolites with antimicrobial effects, such as organic acids, bacteriocins, bacteriocin-like inhibitory substances (BLIS), and others. The biosynthesized metabolites are correlated, on the one hand, to the biochemical characteristics of the lactic acid bacteria and to the composition of the extracts from the studied pseudocereal and legume flours, on the other.Bacillus spp. after the thermal and acid treatments.Similar studies can be found in the literature emphasizing the presence of antifungal compounds produced and released in the culture medium by lactic acid bacteria strains after 48 h of fermentation and stable at pH values ranging from 5.5 to 7.5 . In anotAspergillus niger when the CFS with pH = 7.5 was used. In contrast, other researchers reported the absence of inhibitory effects of some species of Lactobacillus and Pediococcus against Aspergillus flavus and A. carbonarius when a CFS with pH 7.0 was used [Aspergillus flavus and Aspergillus carbonarius were found by Missaoui et al. (2019) [The antimicrobial activity of the lactic acid bacteria is strain dependent although it also depends on the targeted spoilage microorganism and on many other factors such as the fermentation conditions, type of raw materials, etc., thus explaining the diversity of the results found in the literature. Some of the strains isolated in this work are able to biosynthesize compounds which showed a weak inhibition of was used . Nonethe. (2019)  when ferOverall, this research effort presents the selection of lactic acid bacteria strains, in particular, strains no. 2, 24, 30, and 48, which demonstrated valuable biotechnological characteristics in terms of acidification properties, exopolysaccharide biosynthesis, and antimicrobial properties that could be employed in the gluten-free sourdough fermentations. These four strains could be used as a multi-strain starter due to the complexity of their biochemical properties. They have the advantage to be adapted to the gluten-free environment and are rich in protein flours originated from pseudocereals and legumes as fermentation substrates. In future research studies, these selected lactic acid bacteria are likely to be used for the optimization of the biotechnological processes for the production of gluten-free sourdough rich in proteins, based on chickpea, quinoa, and buckwheat flours that have valuable technological and functional properties.The aim of this research work was the isolation and selection of presumptive lactic acid bacteria wild strains, demonstrating important functional properties that can be subsequently used in gluten-free and plant-based protein-rich sourdough fermentations. Such a selection was based on some key technological and functional features for the sourdough fermentation and breadmaking, particularly the capacity of acidification, the capacity to produce exopolysaccharides, and the antibacterial and antifungal activities.Lactobacillus spp. and Leuconostoc spp., have demonstrated better performances in terms of the elected technological and functional features. These strains showed good antimicrobial properties. It should also be mentioned that the lactic acid bacteria strain no. 48 was able to produce high amounts of exopolysaccharides and antimicrobial substances. Nevertheless, strains no. 2 and strain no. 30, presumptively classified as Lactobacillus spp., exhibited the best acidification potential, although they failed, compared to the other two, at the good antimicrobial capacity.Among nineteen presumptive LAB strains isolated from several raw materials, four strains demonstrated beneficial characteristics for gluten-free and plant-based protein-rich sourdough production and were therefore selected for future studies. In fact, strains no. 24 and no. 48, presumptively classified as The reported data is part of a more comprehensive research work that will involve the genotype identification of the isolated microorganisms as well as complementary studies for the phenotype characterization. Furthermore, the future work will involve the optimization of the functional microbial metabolites as well as the optimization of the new sourdough fermentations and formulation for gluten-free and protein-rich plant-based recipes.


Leptospira spp. Here, a serological test was performed on 131,660 pig sera collected between 2002 and 2017 from 4715 farms in Northern Italy. A positivity rate of 13.05% was determined. Australis was the most frequently identified serogroup (77.29%), followed by Pomona (18.47%), Tarassovi (1.51%) and Icterohaemorrhagie (1.40%). Culture isolation and real-time Polymerase chain reaction (PCR) were carried out on 347 kidneys and 470 clinical samples, respectively. Overall, 133 strains were cultured successfully and 43 randomly chosen isolates were identified as serogroup Pomona. Multi-locus sequence typing (MLST) revealed that 41 isolates and 8 DNA extracted from biological samples belonged to sequence type 140. Using a multiple-locus, variable-number tandem repeat analysis, 43 samples produced identical profiles but, after 2014, three new Leptospira interrogans serogroup Pomona genotypes were observed. Interestingly, two isolates showed new MLST profiles and an unclassified identification by monoclonal antibodies. The 16S rRNA gene sequencing clustered them into L. kirschneri species and a core genome MLST analysis revealed an allelic identity of 96% compared with Mozdok strains. Genotyping allowed us to discriminate leptospires and to identify new emerging strains. The accurate identification of infective strains is required for formulating preventive methods and intervention strategies.Swine act as both maintenance and incidental hosts of pathogenic  Leptospira. While, in theory, any pathogenic Leptospira may infect any animal species, leptospirosis is a disease that shows a natural nidality, and each serovar tends to be maintained in specific maintenance hosts. For example, serovar Bratislava is associated with pigs and horses, Canicola is commonly found in dogs, Icterohaemorrhagiae in rats, Pomona in pigs and Hardjo in cattle [Leptospirosis is the most widespread zoonosis worldwide and it is caused by an infection with any of the pathogenic members of the genus n cattle ,2,3. Lepn cattle ,5,6. In n cattle ,5,7. Chrn cattle ,5. In Italy, swine act commonly as carriers for serovar Bratislava, belonging to the Australis serogroup, and serovar Pomona, belonging to the Pomona serogroup. Additionally, serovar Tarassovi causes commonly incidental infections . EndemicUntil 2010, in Italy, a trivalent vaccine against the serogroups Australis, Pomona and Tarassovi was available for swine. Nevertheless, very few breeders have adopted vaccination practices, primarily because the risk of leptospirosis is poorly understood by farmers and because the treatments and/or control strategies for other more virulent diseases are more important to the farm’s economy. In 2011, vaccinations were completely abandoned because no commercial vaccine was available for pigs. Starting from that moment, the main infection control strategy was the management of the breeding herd through the prevention of direct or indirect contact with free-living vectors or other domestic stock. Strong monitoring and surveillance systems are needed to better understand the disease epidemiology, and strict biosecurity tools should be applied to limit the transmission of the infective agent. Leptospirosis is not included in the Office International des Epizooties (OIE) list of notifiable terrestrial and aquatic animal diseases; however, in Italy, it is considered, by the current rules, a notifiable infection . In caseIn the laboratory, serological testing is the most widely used means for diagnosing leptospirosis, and the Microscopic Agglutination Test (MAT) is the serological gold standard method reported by the OIE . IdeallyLeptospira grows slowly over weeks [Leptospira infection has become simpler and faster. Over the years, a number of real-time PCR-based methods have been described for the direct identification of Leptospira nucleic acids in biological samples, both for detection of pathogenic and environmental species [The OIE indicates that strain isolation should be attempted from biological samples. Cultured strains can be studied in depth, and the serogroup and serovar status should be determined using polyclonal and Monoclonal Antibodies (mAbs), respectively . Neverther weeks . With th species ,18,19,20Leptospira, the classical classification into serogroups and serovars has been joined by molecular typing. Several molecular techniques, such as Multiple-Locus Variable-Number Tandem-Repeat (VNTR) Analysis (MLVA) and Multi-Locus Sequence Typing (MLST), aimed at identifying bacteria by examining individual genomic profiles, have been used to investigate the epidemiology of Leptospira. The MLVA method is a useful typing tool for identifying Leptospira genotypes by providing information on genetic relationships among isolates for the surveillance of Leptospira populations. Salaün and colleagues [Leptospira  through the analysis of five loci of repetitive unit sequences .Furthermore, in the last few years, genotypic classification has increasingly integrated traditional serological classification in a wide variety of bacterial species. For lleagues  developeLeptospira, increasing the discriminatory power necessary to distinguish isolates.In 2013, Boonsilp and colleagues  proposedLeptospira in pigs in Northern Italy and to characterize the circulating strains using innovative molecular techniques. Our goal is to understand the epidemiology of swine leptospirosis in this specific area and to identify possible new emerging strains to address preventive and control measures and to reduce the risk of infection in swine herds.This study aims to provide a serological survey of the prevalence of Using MAT, 17,184 out of 131,660 sera collected from 4715 farms were identified as positive, with a seropositivity of 13.05% (cut-off ≥ 1:100). The minimum percentage of positive sera was observed in 2006 (8.46%), the maximum one in 2012 (18.31%). Among the positive samples, 13,809  tested positive for one serogroup, while 3375  were positive for more than one serogroup. Antibody titers against the serogroup Australis occurred in 8.11% of the total samples and 77.29% of the single positive samples, followed by Pomona , Tarassovi , Icterohaemorrhagiae , Sejroe , Ballum , Canicola  and Grippotyphosa  . Using tFor positivity to more than one serogroup , the preThe MAT titers of the single positive samples were generally low (1:100 or 1:200) for all the serogroups detected, except for the serogroup Pomona, which showed medium (1:400) or high titers (greater than 1:800) in more than 50% of the samples . In partLeptospira (Leptospira serogroups. The percentage of farms with positive test results ranged between 22.84% (n = 140) in 2006 and 77.52% (n = 231) in 2012.Out of 4715 tested farms, a mean of 53.62% resulted as positive for the presence of pathogenic ptospira . A farm Outbreaks caused by Australis were stable over the years, with a maximum value of 82.23% in 2012 and a minimum of 66.34% in 2002. This serogroup is responsible for a mean of 75.87% ± 0.15%  of the outbreaks during the study period .The second serogroup in terms of frequency of outbreak was Pomona, with a mean of 14.56% ± 0.31% (CI 95%) over the 16 years of the study with the maximum value observed in 2004 (21.58%) and the minimum one in 2017 (4.17%). Serogroup Icterohaemorrhagiae was responsible for a mean frequency of 3.52% ± 0.30% (CI 95%), with a peak of 8.33% in 2017. In 2017, an inversion occurred between the Pomona and Icterohaemorrhagiae serogroups: the first decreased to the minimum value (4.17%), while the latter increased to a new maximum level (8.33%). The mean frequency of Tarassovi was 2.10% ± 0.48% (CI 95%) but interestingly, its trend started from 11.49% in 2002 and became null in 2014. For Sejroe, a very low frequency was registered over the years, with values between 0% in 2014 and 2.84% in 2016 . For the other serogroups, the mean prevalence was less than 2% with rare exceptions during the considered time .Leptospiral DNA in 102 out of 470 biological samples (21.70%). In total, 133 out of 347 samples (38.33%) were cultured successfully. The MAT carried out using the isolated strains as antigens against reference anti-sera showed that they were antigenically related to the Pomona serogroup. In total, 12 randomly chosen isolates  were subjected to serovar identification with mAbs. The reference strains Pomona and Mozdok 5621 reacted to mAb panels as expected  and st. Mezzano I . In total, 2 (ID 393 and 411) out of 43 isolates showed two new sequence types (STs), similar to ST117, typical of L. kirschneri Mozdok and also previously determined for reference strain 5621 present in the Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER) collection. Compared with ST117 , the glmU gene of sample 393 showed the substitution 120 T > C, and the pntA gene had the substitution 410 A > G. The isolate 411, compared with ST117, showed the mutated pntA gene already found in isolate 393, while the glmU gene was the same as that of ST117 . Both the sequences were submitted to the curators of the Leptospira database and alleles 75 and 85 were assigned to glmU and pntA, respectively. These new alleles defined two new STs, ST288 and ST289, for isolates 393 and 411, respectively.The genotypes of 43 randomly chosen isolates (including the 12 isolates tested for serovar determination) and the 8 biological samples  were detL. interrogans Pomona reference strains (Mezzano I and Pomona) having ST140, while the samples 393 and 411 clustered with L. kirschneri species near the reference strain 5621.As shown in Moreover, the MLVA analysis revealed that 37 out of 41 isolates, belonging to ST140, and 6 out of 8 DNA samples, had the MLVA profile 3-1-10-neg-5 for the VNTR loci 4, 7, 10, Lb4 and Lb5, respectively. This profile was identical to that of the Italian reference strain (Mezzano I), but different from the international reference strain (Pomona) profile (2-0-13-neg-5) at the loci 4, 7 and 10 . Three iL. kirschneri species, in agreement with the previous results.Sequencing the 16S rRNA gene confirmed that strains 393 and 411 belonged to To clarify the genetic features of strains 393 and 411 belonging to the new STs, we performed WGS and a cgMLST analysis. Sequences of strain 393, 411, IZSLER 349/2007 and Mozdok 5621 were submitted to EBI database under the project number PRJEB36553 and are available under genome accession numbers from ERR4056312 to ERR4056315. The cgMLST analysis was performed on contigs of the strains 393, 411, IZSLER 349/2007 and Mozdok 5621 using the Bacterial Isolate Genome Sequence Database (BIGSdb)  Table 6Table 6. The closest cgSTs were queried against the BIGSdb to retrieve deposited isolates with the same genotypes. The results are reported in L. kirschneri Pomona Mozdok . This territory has a very large pig population, with a mean of 5,300,000 animals, and contained 12,900 farms from 2007 to 2017 (data for 2002–2006 are not available), which produce pork for the entire country. In this area, the control of swine pathogens responsible for zoonoses, like Leptospira, is crucial to limit human and domestic animal infections, as well as to reduce the related economic losses linked to such diseases on the farms. Furthermore, the identification of the strains responsible for leptospirosis in pigs and their specific genotypes are essential for epidemiological analyses needed to adopt proper preventive measures and to better understand the etiology of the disease.The main objective of this study was to evaluate the epidemiology of infective In this study, MAT, considered the gold standard and the most commonly used test, was used for serological identification. We tested 131,660 pig sera collected for 16 years. In 2002, there were 18,096 collected samples but the number progressively decreased each year to 2505 samples in 2017, except for a peak of 9330 samples in 2012. This trend may result from the strictness of the Italian national rules , in whicIn the MAT analysis, to discriminate positive and negative sera, we considered a cut-off titer of 1:100, following the guidelines indicated by the OIE  and commConsidering both single positive sera and multiple positive sera, our results indicated a seropositivity of 13.05%, in agreement with other previous studies ,28,29. IOur data revealed that a mean of 53.6% of pig farms sampled in Northern Italy tested positive for leptospirosis, in agreement with the data reported by Bertelloni and colleagues , who obsLeptospira Bratislava, which is widespread among hedgehogs (Erinaceus europaeus) [Leptospira infections, but its epidemiological role remains poorly understood owing to isolation difficulties [L. interrogans Bratislava has only been recovered from swine in a few countries, such as the United Kingdom [The most common serovars associated with swine infections in Italy are Pomona (serogroup Pomona), Tarassovi (serogroup Tarassovi), Bratislava  and Muenchen  . Our resropaeus)  and was ropaeus) . Howeverropaeus) , the Netropaeus)  and Nortropaeus) ,43. Seroiculties . The isoiculties ,46,47, L Kingdom , Germany Kingdom , the USA Kingdom ,49,50, V Kingdom  and Braz Kingdom . Even in Kingdom . In fact Kingdom  found a L. interrogans, with the majority being to Pomona [Leptospira MLST website https://pubmlst.org/Leptospira/ sited at the University of Oxford [Pomona, the second most predominant serogroup in this work, is among the most common serogroup isolated from swine worldwide ,54,55, ao Pomona . Moreovef Oxford . The devFor Tarassovi, historical data supported the widespread presence of this serogroup among the Italian pig population. In 1990, Tagliabue reported the isolation of four Tarassovi strains from swine kidneys , while iAmong the minor serogroups found in tested pigs, we payed particular attention to some of them, discussed below. First of all, serogroup Icterohaemorrhagiae whose low prevalence could indicate the use of good rodent control measures inside and around the tested pig farms . MoreoveL. interrogans). However, we should distinguish two different situations: in the case of isolated strains, since serological analysis by polyclonal anti-sera and by mAbs were possible, we were able to distinguish their serovar (Pomona) and serogroup (Pomona). In the case of DNA samples, we cannot assume that they are definitively serovar Pomona based only on the MLST results. However, we can assume that six of the eight samples belonged to serogroup Pomona serovar Pomona because their MLVA profiles were identical to that of strain Mezzano I. Through genotyping techniques we found that all the isolated and extracted samples belonged to the Pomona serogroup but, interestingly, from 2014, new Pomona genotypes started to circulate in Northern Italy. Unfortunately, the Sequence Type provided by MLST analysis (ST140) being associated with isolates characterized as belonging to serogroups Pomona serovar Pomona, serogroup Pyrogenes serovar Guaratuba and serogroup Grippothyphosa (data from BIGSdb) did not permit any discrimination at the serogroup and at the serovar level, this ST In this case, the information provided by MLST genotyping is limited to species discrimination  in 1962, in the Plovdiv District of Bulgaria by Ivanov and later found in rodents in some parts of Europe [L. kirschneri Mozdok st. 5621. In addition, the cgMLST analysis revealed that the most similar strains are classified as Leptospira, species kirschneri, serovar Mozdok. Definitely, MLST and cgMLST analyses excluded their belonging to the Tsaratsovo serovar, because strain Tsaratsovo B 81/7 was characterized as ST115 and cgST 245. We can also exclude Altodouro and Kunming, the other serovars included in L. kirschneri species and the Pomona serogroup, because they showed ST100 and ST70, respectively, with none of the seven loci being in common with those of ST288 and ST289. Therefore, we can assert that they are very similar to L. kirschneri Mozdok, but accurate descriptions of their genetic contents in comparison with serovar Mozdok are still being compiled. From an epidemiologic point of view, several studies have identified serovar Mozdok across Europe [Interestingly, two isolates showed new mAb responses and new STs. Their reactions to the mAbs specific for the Pomona serogroup were very similar to each other but different from any of the known serogroup Pomona strain profiles. Their pattern was compatible with serovar Tsaratsovo, which was first isolated from harvest mice , Pomona (Pomona) and Hyos (Tarassovi) was available for swine. It was used in breeding boars, in non-pregnant sows and in pregnant sows before the octave week of gestation. An unpublished survey carried out by the National Centre of Leptospirosis of IZSLER revealed that among 333 pig farms in Northern Italy during the period February 2004–December 2005, only 19% of the sampled farms adopted the vaccination practice. The reasons for this limited use of vaccination against leptospirosis in pigs included the risk that this disease is poorly understood by farmers and that leptospirosis is a secondary problem in the farm life/economy compared with other more virulent and economically important diseases. Nevertheless, since MAT does not discriminate between vaccination titers and titers due to exposure and because the vaccination status of pigs tested in this study prior to 2010 were unknown, the risk of having an altered number of positive pigs due to the interference of vaccination should be considered. About this aspect, the previously mentioned survey reported that in the farms where vaccination against Leptospira was practiced, 41% of pigs resulted as seropositive, while where it was not practiced seropositive animals were equal to only 9.8%. However, when considering this situation, it highlighted various scenarios: in the case of Australis, the interference of vaccination immunity was significant because the percentage of seropositives increased from 6.1% (of unvaccinated farms) to 33.9% (of vaccinated farms), while for Pomona and Tarassovi, the interference level was lower, going from 2.4% to 5.4% and from 1.2% to 5.4% for unvaccinated and vaccinated farms, respectively (data not published). Based on these data, we can exclude the interference of vaccination on our seropositivity results, given that the overall seropositivity and the seropositivity levels related to Australis, Pomona and Tarassovi  were in agreement with the data from the unvaccinated farms reported in the survey . Furthermore, in this study the trends in the seropositivity of Australis, Pomona and Tarassovi did not reveal any remarkable changes after the end of vaccine use in Italy occurred in 2010, which provided further evidence for our statement. Our study, which provides information on the Considering the serological data obtained and the frequency, in particular, of the Australis and Pomona serogroups, the use of vaccination in pig herds is highly advisable. In this regard, in March 2019, a vaccine has been registered in Italy. This vaccine contains nine serovars  and is active against six serogroups . As reported by Jacobs et al. , this vaLeptospira infections was previously included in the disease-monitoring programs. The samples were collected and analyzed by the National Reference Centre for Animal Leptospirosis at the Istituto Zooprofilattico Sperimentale della Lombardia ed Emilia Romagna (IZSLER) located in Brescia . In this study, data already published by Tagliabue [A total of 133,660 sera, 347 kidneys and 470 biological samples  were included in this study. These samples were collected over 16 years (from 2002 to 2017) from 4715 pig farms located in Northern Italy (Lombardy and Emilia Romagna regions) during the routine diagnostic activity. In these farms, testing for agliabue  collecteStarting from 2008, 347 well-preserved samples were submitted for culture-based causal agent isolation. Since 2009, when molecular methods for the routine diagnosis of leptospirosis were introduced, 470 clinical samples were analyzed using a LipL32-based real-time PCR assay.To cover the entire analysis period, 43 randomly isolated samples and DNA extracted from 8 biological samples were submitted to genotyping analysis . For theLeptospira using the MAT in accordance with OIE standards [Leptospira were used (Sera were examined for the presence of antibodies against pathogenic tandards . For MATere used . They weLeptospira interrogans Pomona). The assay was performed on a Bio-Rad CFX96 System using the following thermal conditions: a holding stage of 95 °C for 5 min, and 45 cycles of 95 °C for 15 s and 60 °C for 30 s. DNA was extracted from 0.5–4.0 mL of urine or tissue homogenate using the PureLink Genomic DNA kit  according to the manufacturer’s instructions. An internal control DNA (0.1 µL per µL of elution volume) was added to the digestion buffer. A Taqman-based PCR assay targeting the lipL32 gene was used to detect pathogenic leptospires using primers described previously . The PCR−2 to 10−5) performed in semisolid selective EMJH (added with 5-fluoruracil and agar). The cultures were incubated at 30 ± 1 °C and observed under dark-field microscopy weekly for up to 6 months. In case of contamination, the cultures were filtrated through a 0.22 μm sterile syringe filter and sub-cultured in fresh semisolid selective EMJH medium.For isolation, 1 g of tissue was diluted, in a plastic bag, in 9 mL of EMJH added with 5-fluoruracil (selective EMJH) ,80 and hThe serogroups of the isolates were determined by MAT using a panel of eight polyclonal anti-sera against the eight serovars described in L. interrogans Pomona st. Pomona and L. kirschneri Mozdok st. 5621, provided by KIT, were used as controls for serovar identification. To interpret the results, we referred to the expected maximum dilution titer provided by the Leptospirosis Reference Centre of KIT, as reported in Serovar classification was achieved by MAT using panels of mAbs directed against polysaccharide antigens, specific to the serogroup identified. A battery of five mAbs , which react with serovars of the serogroup Pomona ,83, were® Gel and PCR Clean-up  or with exonuclease I and FastAp Thermosensitive Phospahatase alkaline  according to the manufacturer’s instructions. Cycle sequencing reactions were performed using the BigDye® Terminator Cycle Sequencing kit version 1.1 . Reactions were purified using a BigDye® XTerminatorTM Purification kit Thermo Fisher Scientific, Waltham, MA, USA) and sequenced on a Genetic Analyzer 3500xl sequencer (Thermo Fisher Scientific) according to the manufacturer’s instructions. Nucleotide sequences were assembled using the SeqMan module of the Lasergene sequencing analysis software package . Assembled sequences were trimmed and aligned to allele reference sequences downloaded from the Bacterial Isolate Genome Sequence Database (BIGSdb) [https://pubmlst.org/Leptospira/), to assign allele numbers to all seven loci. For strain identification, allelic profiles were queried against the Leptospira BIGSdb.To genotype leptospires we used the scheme proposed by Bonsilp in 2013 , based o(BIGSdb)   were used to characterize the isolates and the extracted DNA samples as described by Salaün and colleagues in 2006 . BrieflyLeptospira isolates from pigs belonging to new Sequence Types (ID 393 and 411) were extracted from the liquid EMJH cultures using the DNeasy Blood and Tissue kit (Qiagen), according to the manufacturer’s instructions. Extracted DNA was successfully used for species confirmation and for WGS. One strain isolated from pig previously identify as L. interrogans servar Pomona, “strain IZSLER 349/2007”, and the reference strain Mozdok 5621 were included as controls for the cgMLST analysis.Genomic DNA of two Leptospira strains.The species assignments of the isolates 393 and 411 were confirmed by sequencing portions of their 16S rRNA genes using the Microseq 500 16S rDNA PCR kit . The obtained sequences were compared with a controlled and validated reference database  and through BLAST-based analyses to determine the species of the Leptospira contigs were used for further analyses. The draft genomes of samples 393, 411, IZSLER 349/2007 and strain Mozdok 5621 were used to perform cgMLST analyses with BIGSdb [WGS was performed on the four genomic DNAs extracted from cultured strains  using an Illumina NextSeq  platform with a paired-end 150× 2-bp run starting from a genomic library prepared with a Nextera DNA Flex kit (Illumina). Raw reads were filtered using Trimmomatic ver. 0.38 , assemblh BIGSdb , using th BIGSdb .Ethical statement: The study was exempt of ethical approval procedures because animal samplings were performed during the routinely diagnostic procedures in naturally infected farms.L. interrogans serogroup Pomona had been circulating for 10 years (from 2002 until 2013) and interestingly, L. interrogans strains collected from 2014 onwards had various heterogeneous genetic profiles. Furthermore, new strains belonging to L. kirschneri species have been identified for the first time. Since an accurate identification of the infective strain is crucial for an adequate vaccine formulation, these findings provide an important contribution for addressing prevention and intervention strategies to reduce infection risks on farms.In this study, an intensive serological survey evidenced that Australis and Pomona were the most serogroups causing leptospirosis in pigs in Italy. Molecular analyses revealed that a stable genotype of strains belonging to


Biomedical waste management is getting significant consideration among treatment technologies, since insufficient management can cause danger to medicinal service specialists, patients, and their environmental conditions. The improvement of waste administration protocols, plans, and policies are surveyed, despite setting up training programs on legitimate waste administration for all healthcare service staff. Most biomedical waste substances do not degrade in the environment, and may also not be thoroughly removed through treatment processes. Therefore, the long-lasting persistence of biomedical waste can effectively have adverse impact on wildlife and human beings, as well. Hence, photocatalysis is gaining increasing attention for eradication of pollutants and for improving the safety and clearness of the environment due to its great potential as a green and eco-friendly process. In this regard, nanostructured photocatalysts, in contrast to their regular counterparts, exhibit significant attributes such as non-toxicity, low cost and higher absorption efficiency in a wider range of the solar spectrum, making them the best candidate to employ for photodegradation. Due to these unique properties of nanophotocatalysts for biomedical waste management, we aim to critically evaluate various aspects of these materials in the present review and highlight their importance in healthcare service settings. Biomedical waste management has recently risen as one of the major challenges that developing countries are confronting. The amount of biomedical waste produced has considerably increased as the worldwide populace has expanded, and accessible assets are not sufficient to deal with it . DisposaSuccessful and efficient management of biomedical waste requires the use of different treatment practices and techniques, such as incineration, autoclave, hydroclave, and microwave treatments . It is eThe other commonly used methods to treat biomedical waste are mechanical treatments such as granulation, pulverization, shredding, grinding, mixing, agitation, and crushing. This type of treatment has the advantage of reducing the bulk volume of the waste materials by 60 percent or more. Although mechanical treatment does not remove the pathogens or disinfect equipment, it reduces the waste volume to facilitate further treatment or disposal. Equipment involved in mechanical treatment includes but is not limited to crushers, millers, shattering machines and splinterers. These treatment methods can alter the appearance of the waste, which can be useful in reducing the psychological impact of the waste on human observers. In addition to reducing the volume of bulk disposal, mechanical treatment can increase the surface area of the solid pieces before subsequent chemical or heat treatment.Chemical disinfection, such as through the use of chlorine compounds, has been widely used to eliminate the microorganisms in medical waste, as well as oxidizing hazardous chemical constituents. For instance, chlorine bleach has been used to disinfect swimming pools and reduce the risk of disease transmission. Another example of chemical disinfectant compound is ethylene oxide treatment, which is used to disinfect materials and sometimes to treatment of medical waste. Ethylene oxide (EtO) treatment is used to sterilize the equipment that will be frequently used. This disinfectant chemical is not cost effective for use on equipment or treatment of waste that will be disposed of in a landfill. EtO gas can be used to kill microorganisms and disinfect products during packaging processes.Microwave radiation has recently been employed to treat wastewater sludge and to generate heat for treating medical waste. This method of waste treatment can be employed either on-site or mobile by using treatment vehicles. To enhance the efficacy of the microwave treatment and reduce the volume of the final product, the waste will go through a shredding process first. In the case of using dry waste, the waste is wetted with water and then introduced into the microwave chamber, as this method of disinfection is effective only when the waste is damp. Therefore, the microwave treatment units are usually supplied with a humidifier. Although the whole disinfection time is determined by the manufacturer and experience of the operators, approximately 20 min per each batch is required. Microwave should not be confused with irradiation such as gamma rays (from radioactive elements) or electrons, as these two methods are completely different.Gamma irradiation is a means of sterilization by exposing waste to gamma rays, as it breaks down bacterial DNA. To generate gamma rays, radioactive isotopes of cobalt are employed, this is the same radiation source used for the radiation treatment of cancer. However, in cancer treatment, the radiation is intended to kill the malignant cells, whereas to sterilize equipment or treat waste, pathogens are targeted. In contrast, the ultraviolet (UV) radiation used to treat wastewater is not capable of killing microbes so much as it is able to break down chemicals. The efficiency of irradiation as a means of sterilization is highly dependent on the total energy delivered, but even then, this method of treatment suffers from the shadowing effect, which means that waste surfaces facing the radiation source are more sterile than the waste on the shaded side. Therefore, waste with odd shapes, and the sides of contaminated surfaces facing away from the cobalt source, may not be adequately exposed to the radiation. Heat treatment, by contrast, brings every piece of waste to an adequate temperature for sterilization, if done properly.Although vitrification—the means production of glass—has rarely been used, it could be an effective treatment for medical waste. The high temperature kills pathogens and some combustible material via burning or pyrolysis, which results in an off-gas. The remaining by-product is encapsulated in glass, which has a very low diffusivity. However, this method of disinfection might become dangerous if significant quantities of the encapsulated hazardous material leach out of the glass. Ultimately, the vitrified waste can be disposed of in a landfill with confidence. Despite the development of plasma treatment as an alternative to incineration for medical waste treatment, it has not been widely implemented .To decrease the expense of waste treatment through cost-effective strategies, various methods based on the exploitation of sunlight have been proposed for both solid and liquid waste management . Among tThe optimal treatment system depends on many factors, such local conditions, availability of resources including technical expertise, waste characteristics and volume, relevant national regulations and safety requirements, technical requirements for installation, operation and maintenance of the treatment system, environmental factors and cost considerations. Nonetheless, waste management systems can be changed and improved only within the financial and technical capacity of a given health-care system, which may then require making small decisions towards an incremental improvement, as well as planning for the attainment of long-term improvements, once certain conditions have been met.Nanophotocatalysts have been widely used to treat waste in the field of environmental and ecological safety, as they have numerous benefits, such as those of low cost, superb stability, high photocatalytic activity, innocuousness to humans, etc. . DiffereAlthough approaches to removing pollutants based on nanostructured catalytic membranes, nanosorbents and nanophotocatalyst are eco-friendly and efficient, in order to purify the waste, they require more energy and sufficient investment. There are many challenges involved in biomedical waste treatment; some precautions are required to keep hazardous waste away from ecological and health issues. New modern equipment for waste treatment is required to be flexible, low cost and efficient for commercialization purposes. Recently, with advancements in nanomaterials such as nanophotocatalysts, nanomotors, nanomembranes, nanosorbents and imprinted polymers, the decontamination of biomedical waste has been effectively revolutionized.However, there has not been a systematic characterization of the risk and hazards related to nanomaterials, and there is a lack of safety regulations for using such catalysts. Overall, nearly all nanocatalysts have toxic effects both in vitro and in vivo at certain concentrations. For instance, ROS generation and cell signaling perturbations are widely accepted causes of nanotoxicity. Furthermore, the toxicity of nanoparticles is also determined by factors such as particle size and surface functionalization . AlthougThis review aims to introduce different types of nanophotocatalysts and provide the main principles, mechanisms, and operating parameters of the photocatalysis process and emphasize its importance in biomedical waste management in detail see .2 and OH (oxidation). The kind of ROS depends on the type of material and irradiated photons. A conventional photocatalysis procedure is demonstrated in Photocatalysis is defined as a series of chemical reactions that is usually initiated by electromagnetic irradiation. The photocatalysis process can be divided into two main stages of reduction and oxidation. When a material is irradiated with photons with energy equal to or higher than its bandgap, the excited electrons in the conduction band (CB) will jump to the valence band (VB) through the bandgap leaving positive holes, which is called reduction. As a result, the generated electrons and holes lead to the formation of reactive oxygen spices (ROS) such as OIn the photocatalytic process, the catalyst and light are simultaneously used to accelerate a chemical reaction. Thus, photocatalysis can be defined as the catalysis-driven speed-up of a light-actuated reaction. Photocatalysts are categorized into two classifications: hetero- and homogeneous procedures . When coThe influence of different factors on photocatalytic efficiency has been investigated and further optimized using the response surface methodology (RSM). The RSM is a combination of both statistical and mathematical methods to optimize a complicated process. To optimize the process more accurately, this statistical design of experiments considers the interaction effects between the studied parameters, and can determine the combination of levels. Furthermore, a central composite design (CCD) based on RSM can be successfully applied in the optimization of photodegradation of various organics .Since the adsorption of pollutants on the surface of solids plays a critical role in heterogeneous photocatalysis, the effect of adsorption on photocatalytic process has been investigated. Simultaneous adsorption and photocatalytic processes has been used for degradation of phenol pollutants with noticeable recyclability and stability of the photocatalyst , bacteriNano-sized photocatalyst particles demonstrate a significantly intensified reactivity compared to larger particles or bulk materials due to their large surface area ,30. NoveMetal-free catalysis, such as with graphitic carbon nitride nanocomposites, has also been used in photodegradation of aqueous phase organic pollutants ; likewis2, ZnO, α-Fe2O3, and WO3 are the most suitable ones, since they are photocorrosion resistant and have a wide band gap energy. TiO2 is currently used as the most efficient photocatalyst, and is widely applied in wastewater treatment, since it can promote the oxidation of organic compounds while being thermally stable, non-toxic, cost-effective, and chemically and biologically inert. Structural and surface properties, including surface area, porosity, crystal composition, particle size distribution, and band gap energy, are able to affect the photocatalytic activity of the catalyst [Among different kinds of photocatalysts, metal oxide semiconductors, including TiOcatalyst . The folcatalyst . First, catalyst .Moreover, changing the size of the nanocatalyst makes it possible to adjust the absorbance wavelength. Additionally, the optical and electronic characteristics of the nanocatalyst can be modified by tuning their sizes . Due to The efficacy rate in photocatalysis methods is primarily dependent on the size, shape, crystal structure and surface area of the photocatalyst, as well as the morphology, which can also act as a significant factor affecting the final degradation throughput. The amount of catalyst is also directly proportional to the overall rate of photocatalytic reaction. Disinfection efficiency of the photocatalyst can be improved by an increase of its doses . The pH As photocatalysts have a superior capability in the deactivation of various destructive microorganisms, they could reasonably be used as alternatives to conventional techniques , which can generate harmful and undesirable by-products . PhotocaHospital waste can be described as a combination of both biological and non-biological waste that is discarded and not intended to be used again. Hospital and laboratory waste can be broadly defined into two categories: hazardous (risk waste) and non-hazardous (non-risk) waste . Risk waThere are two main steps involved in the waste disposal process: treatment and final disposal. In the treatment step, depending on the nature of the waste, procedures such as incineration, autoclaving, chemical disinfection, encapsulation, and microwave irradiation are usually applied. The disposed wastes are likely to wind up in landfill, buried inside the premises, discharged into the sewer, etc. Segregation (separation) has a key role in an efficient waste identification and management process. Sorting the waste based on the color of the container is the most proper method for identifying waste categories . The lasStaphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), Enterococcus faecium (E. faecium), and Escherichia coli (E. coli) [Photocatalysis is an efficient, affordable strategy for the decontamination and disinfection of hospital waste that applies ultraviolet rays or solar energy to disintegrate antibiotics and disinfect microbes from the waste at the point of origin see . In demaE. coli) .E. coli (ATCC8739) cells on membrane filters during fluorescent light irradiation [2 thin films on E. coli [Bacillus pumilus (B. pumilus) and has shown higher photocatalytic activity compared to the glass substrates coated with self-sterilizing materials [Another example of self-disinfecting surface application is the inactivation of the deposited adiation . In anot E. coli . Due to aterials .E. coli [2 photocatalysts have been used owing to their visible light-induced bactericidal activity against human pathogens [E. coli using nitrogen/sulfur-co-doped Titania [Surfaces coated with CuO-doped Titania photocatalysts were also assessed for their biocidal activity and synergistic impact of photocatalysis and lethality of copper to deactivate bacteriophage T4 and E. coli . Nitrogeathogens ,70. Visi Titania ,62,63,71 Titania ,73.Legionella bacteria, is another application of photocatalysts in a laboratory or hospital [In another study, prions, an infectious agent of a group of transmissible, fatal neurodegenerative disorders influencing both animals and humans, were inactivated using photocatalysts . Photocahospital .2/UV photocatalytic oxidation has the capacity to mineralize the four strains of L. pneumophilia serogroup 1 cells  in laboratory-scale surveys, which proves that this could be used as a reasonable procedure for the routine disinfection and sterilization in order to control Legionella bacteria species in the hot water systems of emergency clinics and hospitals  [2 as an effective treatment has also been reported to remove pharmaceutical contaminants from water with high efficiency. Due to their high photolysis sensitivity in aquatic systems, they have been used, for instance, to treat diclofenac [It has also been shown that TiOication) . The appclofenac ,75, and clofenac .Biomedical waste refers to any type of waste generated during treatment, diagnosis, or research activities—testing biological products and the immunization of human beings or animals. Biomedical waste management has become a general medical issue, as it is not only a legal necessity, but also a social responsibility. An improvement of biomedical waste management starts with the reduction of the waste produced .According to an earlier report, around half of the world’s population is in danger of incompatible biomedical waste management, which can impact both work and public places . About 7The essential needs in biomedical waste management include reducing, waste storage and transport, recycling and reusing, expenses in the annual budget, storage management, separate chemical and pharmaceutical waste segregation, separate storage zones, and documentation related to biomedical waste management . Waste tStaphylococcus aureus (S. aureus), a typical pathogenic bacterium in implant-related infection, shows photocatalytic activity using TiO2 film on titanium substrates and stainless steel [2 coatings have been used on bioimplants to apply photocatalysis for antibacterial purposes. The coatings exhibited a bactericidal impact upon UV irradiation, so the implementation of these photocatalytic-coated substrates are a valuable system for controlling infections related to biomedical implants [Pseudomonas aeruginosa, E. coli, Enterococcus faecium, and S. aureus) [Photocatalytic processes are used in biomedical applications due to their disinfection abilities. It has been investigated that ss steel . TiO2 coimplants ,65,66. Aimplants . Reducinimplants . A signiimplants . The hig aureus) ,90, and  aureus) ,91,92.2, ZnO, α-Fe2O3, MoO3, WO3, ZrO2 and SnO2 can be applied as photocatalysts, which are of great interest in current studies [2)Titanium Dioxide  structured TiO2 nanophotocatalyst can be replaced with TiO2 nanoparticles by which both the safety level and efficiency of purification of the final purified water are preserved. These structures are suitable for environmental and biomedical applications, as they meet the human key safety conditions, according to an in vitro cytotoxicity test of well-purified water by eco-TiO2 [2 photocatalytic degradation of organic and inorganic pollutants are summarized in TiOompounds , dyes inompounds , toxic compounds , naturalompounds , and a sompounds . Anotherompounds . It has scalpels , surgicascalpels , and catscalpels . The UV-implants ,106. Thel fields . Conventeco-TiO2 ,108. Var2.Zinc Oxide (ZnO)2 [2), ZnO can form stable, protective coatings, which act as smart materials. Three different crystalline phases of ZnO include zinc-blende, wurtzite, and rock-salt. Due to these multi-functional qualities, ZnO is extensively used for various applications including photocatalysis [2 nanoparticles, which are discussed in the binary metal oxide category, have been applied to remove methylene blue and acridine orange [ZnO shows efficient activity in photocatalytic degradation of organic contaminants as compared to TiO2 . Among datalysis , light-eatalysis , biosensatalysis , field-eatalysis . Differeatalysis , nanonaiatalysis ,116, nanatalysis , nanotubatalysis ,119, nanatalysis , nanoribatalysis ,122,123,atalysis , and nanatalysis . It has atalysis ,127. Phoatalysis . In anotatalysis . Additioe orange .3.2O3)Iron Oxide  exhibits desirable efficiency as an important photocatalyst [2O3 has been investigated. However, the corresponding photocatalytic mechanism has not been described in detail. The valence electrons of Fe2O3 compared to those of TiO2 can be excited to the conduction band at wavelengths shorter than 560 nm, which can extensively enhance the efficiency of the sunlight use. The maximum degradation efficiency of 94% for dibutyl phthalate in wastewater  was obtained using Fe2O3 in a photocatalytic process [2O3 powders, porous α-Fe2O3 films exhibit better photocatalytic activity by water splitting under UV radiation for hydrogen generation [2O3 under UV irradiation and natural sunlight in aprotic and protic solvents has also been reported [Iron oxides play an important role in many biological and geological processes. They are increasingly applied as pigments, iron ores, catalysts, and as hemoglobin in the blood. Freely dispersed bulk-, sonic-, and nano-Feadiation . Iron oxcatalyst , with locatalyst  with no catalyst . Due to catalyst ,135,136. process . Compareneration . Photocareported .4.2O3)Gadolinium Oxide (Gd2O3-modified bismuth vanadate (BiVO4) composite, as a photocatalyst, exhibits significantly greater visible-light photocatalytic activity than pure BiVO4 for methyl orange degradation under visible light irradiation [2O3 nanorods used to detect ethanol by facile hydrothermal routes demonstrated a lower detection limit with higher sensitivity and shorter response time [2O3 nanostructures [2O3 hollow microspheres [2O3 photocatalyst prepared by the sol–gel method was measured after 4 h of UV light irradiation [2O3 nanorods was designed for the degradation of neurotoxic chloramphenicol drugs [The global interest in using rare earth metals is increasing, due to their distinctive magnetic and electronic attributes in the fashioning of interfaces and surfaces compared to common bulk materials. Gadolinia (gadolinium (III) oxide) is the most widely available derivative form of gadolinium, and is a potential contrast agent in magnetic resonance imaging (MRI). The Gdadiation . Gd2O3 nnse time  comparedructures . Moreoveospheres . The degadiation . In anotol drugs .5.2O4)Antimony Oxide  and Sb (V). Antimony has been applied as a pacifier in enamel, flame retardants, paint and glass art crafts and for making bullets and bullet tracers. It has been used as an alloy for the synthesis of plain bearings, batteries, and solders, as well as as a stabilizer and a catalyst for the preparation of polyethylene terephthalate.2O4 has been demonstrated, with almost 52% degradation of acridine orange in 170 min, with low detection limit, good sensitivity, long linear dynamic range with good linearity in a very short response time [The photocatalytic activity of α-Sbnse time , as wellnse time , while inse time . The uninse time .3, ZrO2, ZnO, and Fe2O3) that have been exploited in photocatalysts for the degradation of organic contaminants have inherent drawbacks [3 is a stable photocatalyst for O2 production within the visible light irradiation range. However, it is not suitable for H2 evolution because of its low level of conduction band. Additionally, α-Fe2O3 is somewhat stable in acidic solutions, but has the same problems as WO3. Moreover, ZnO can be easily corroded under band gap irradiation by photogenerated holes.In addition to metal oxides, some other metal oxides have also been studied previously for use in the field of photocatalysis, because of their unique benefits and wide range of applications as catalysts, semiconductors, superconductors, ceramics, antifungal agents, adsorbents, and their specific applications in medicines. Many metal oxide semiconductors . However, photo-corrosion, which is a common issue in most metal sulfide photocatalysts, occurs when using both CdS and ZnS. Therefore, recent studies have focused on the development of ZnS and CdS photocatalysts, mostly through four different means of improvement: matrixing and supporting the structures of CdS, adding cocatalysts to CdS, preparing porous and one-dimensional CdS, and doping solid solutions of CdS and ZnS .To synthesize porous CdS, a solvothermal method has been used to synthesize CdS nanowires  and nano2O3), magnetite (Fe3O4), cobalt ferrite (CoFe2O4)) and a photocatalytic shell  [2O4 have shown desirable degradation efficiency of organic target compounds under both visible light and UV irradiation [3O4 [2O4 [2O4 [2O4 [12O19 [12O19 [2 nanophotocatalysts  . Furtheradiation . Similarion [3O4 ,166,167,3O4 [2O4 , CoFe2O42O4 [2O4 , ZnFe2O42O4 [2O4 , BaFe12O4 [12O19 , SrFe12O9 [12O19  -doped Tatalysts . A schem2–G [2–G [2WO6–G [2O4–G [4–G [Graphene (G), due to its one-of-a-kind nanostructure and particular properties has been studied widely from both experimental and theoretical scientific points of view . It has 2–G ,178,179,2–G [2–G , Bi2WO6– [2WO6–G , ZnO–G [ [2WO6–G , ZnFe2O4G [2O4–G , BiVO4–G4–G [4–G , CdS–G [4–G [4–G  have alsQuantum dots (QDs), as zero-dimensional semiconductor multifunctional nanomaterials have been receiving significant attention for the degradation of pollutants . Since Q2 and ZnO due to their low cost, high stability and strong capacity for the photocatalytic decomposition of organic contaminants [2 has been used recently to make self-decontaminant textiles that offer high antibacterial activity performance for UV shielding [Smart photocatalytic materials have been developed widely over the past two decades . Differeaminants . TiO2 hahielding . Nanophohielding  or onto hielding .2-coated membranes offer outstanding antifouling/self-cleaning, photoactive, and bactericidal properties that are based on the UV mechanism of TiO2 photocatalysis . The recent advancement in the field of photocatalysis technology is investigating novel agents with higher photocatalytic performance to expand their light respond range. To address these issues, coupled semiconductor, noble metal deposition and ion modification, are the proposed methods to improve energy band and photocatalytic efficacy of explicit applications. In addition, due to concurrent photocatalytic and redox reactions, sensible photocatalytic systems can be designed for the simultaneously photocatalytic treatment of two or more contaminants [Biomedical waste eradication strategies are still very limited due to low photocatalytic efficacy. Therefore, extensive assessments are highly demanded from the practical point of view . The appaminants . Howeveraminants . To imprThe implementation of the photocatalytic method for disinfection and cleansing is an adaptable and effective procedure for incapacitating a broad range of adverse microorganisms in different media. This approach is a non-toxic, safe, and cost-effective sterilization technique whose versatility enables it to be used in various purposes. Nanotechnology has shown an incredible potential to improve the effectiveness of biomedical waste treatment . TherefoThere is an ongoing research in field of nanomaterials to design and develop nanophotocatalytic reactors. Despot of the advancements in field of nanophotocatalytic materials, further investigations are required to be done to characteristics the nanophotocatalytic materials. The major remaining challenges are strengthening the process, mass transfer limitations and high photons consumption. Therefore, the concept of using nanocomposites is ideal to resolve the issues related to electron pair recombination which can be prolonged by combining the nanocomposites with nanophotocatalytic reactor structures. The recent reactors known as microfluidic reactors open a new opportunity for intense characteristics study in reaction and synthesis phase. Microfluidic reactors are based on micro level reactants. The remarkable features of these reactors are; improved diffusion effect and great mass transfer coefficient factor, large surface-to-volume ratio, highly stable hydrodynamics, less Reynold’s flow, and informal handling which make them better candidate compared to the conventional reactors. However, implementing the photocatalysis in a larger scale and actual wastewater is still challenging. Synthesis of structures such as nanorod, nanosphere, nanoflowers, nanoflakes and nanocones with improved functional and structural properties could open a new area of study in this subject. Crucially nanophotocatalysts with excellent efficiency, inexpensive, eco-friendly and high stability are needed to be synthesized .Since the outbreak of the coronavirus (COVID-19), its impact upon human health and the economy has been increasing day by day; administrations are advised to treat waste management, including of medical, household and other harmful waste, as an urgent and crucial public service in order to reduce the possible risk of secondary impacts upon public health and the environment.During such an epidemic, various types of medical and hazardous waste are generated, including infected personal protective equipment (PPE) masks, gloves and other protective equipment, as well as a higher volume of non-infected items of the same nature that can easily become mixed with domestic garbage, but should be treated as hazardous waste and disposed of separately from other household waste streams. Unreliable waste management could lead to unexpected “knock-on” impacts on human health and the environment. During the COVID-19 emergency, safe management of household waste is also expected to be critical. To overcome this enormous and unprecedented challenge, decision-makers are urged to make every effort to ensure that waste management, including that from medical and household sources, is given the attention—indeed priority—it requires in order to minimize its impact upon human health and the environment.The COVID-19 outbreak has made a global demand to effectively diagnose, treat and mitigate the spread of the infection, through comprehensive approaches such as specific alternative antiviral methods and classical disinfection protocols. The physicochemical properties of materials can be engineered to offer distinctive approaches to manage this emergency. Considering the life cycle of the virus, it is envisioned that nanotechnology could be employed to encounter the disease; nanoparticles (NPs), for instance, can be used as an alternative to the conventional disinfection protocols used in the healthcare system, owing to their inherent antipathogenic properties and their ability to inactivate viruses, bacteria, fungi, and yeasts either through photothermal or generation of photocatalysis-induced reactive oxygen species (ROS). In conclusion, nanotechnology can play a critical role in counteracting COVID-19 and preparing for future pandemics .