Abstract: Small heat shock proteins (sHsps) are ubiquitous conserved chaperone-like proteins invo... more Abstract: Small heat shock proteins (sHsps) are ubiquitous conserved chaperone-like proteins involved in cellular proteins protection under stressful conditions. In this study, a reverse transcription quantitative PCR (RT-qPCR) procedure was developed and used to quantify the transcript level of a small heat shock gene (shs) in the probiotic bacterium Lactobacillus acidophilus NCFM, under stress conditions such as heat (45 °C and 53 °C), bile (0.3 % w/v), hyperosmosis (1 M and 2.5 M NaCl), and low pH value (pH 4). The shs gene of L. acidophilus NCFM was induced by salt, high temperature and acidic stress, while repression was observed upon bile stress. Analysis of the 5 ' noncoding region of the hsp16 gene reveals the presence of an inverted repeat (IR) sequence (TTAGCACTC-N9-GAGTGCTAA) homologue to the controlling IR of chaperone expression (CIRCE) elements found in the upstream regulatory region of Gram-positive heat shock operons, suggesting that the hsp16 gene of L. acidoph...
Abstract: Currently, the majority of prebiotics in the market are derived from non-digestible oli... more Abstract: Currently, the majority of prebiotics in the market are derived from non-digestible oligosaccharides. Very few studies have focused on non-digestible long chain complex polysaccharides in relation to their potential as novel prebiotics. Cereals β-glucans have been investigated for immune-modulating properties and beneficial effects on obesity, cardiovascular diseases, diabetes, and cholesterol levels. Moreover, β-glucans have been reported to be highly fermentable by the intestinal microbiota in the caecum and colon, and can enhance both growth rate and lactic acid production of microbes isolated from the human intestine. In this work, we report the effects of food matrices containing barley β-glucans on growth and probiotic features of four Lactobacillus strains. Such matrices were able to improve the growth rate of the tested bacteria both in unstressed conditions and, importantly, after exposure to in vitro simulation of the digestive tract. Moreover, the effect of β-gl...
Fermented oat-based foods offer attractive prospects within the market of non-dairy functional pr... more Fermented oat-based foods offer attractive prospects within the market of non-dairy functional products, since they are suitable substrates for the delivery of probiotic microorganisms, and are significant sources of dietary fiber, both insoluble and soluble such as β-glucan, good quality fat and other phytochemicals important for human health In the present work, whole oat flour was fermented with probiotic Lactobacillus plantarum strains to produce new functional foods with improved nutritional and technological features. Viability of the probiotic and the main technological, physico-chemical, nutritional and sensorial parameters were monitored at 7, 14 and 21 days of cold storage. The microbial survival was higher than 5x10 8 cfu g-1 at the end of the shelf life. After the fermentation step, viscosity was higher in products inoculated with the exopolysaccharide-producing L. plantarum strain Lp90. However, a subsequent viscosity reduction was detected in all the samples throughout the storage period, consistent with the observed concentration decrease of the oat β-glucan. Vitamin B2 content was about twofold higher in products fermented by the riboflavin-overproducing LpB2, and in these samples the riboflavin concentration further increased during cold storage. Highlights Fermented oat-based foods are suitable substrates for delivery probiotic LAB. Riboflavin-overproducing LpB2 increased the vitamin B2 content of oat-based products. EPS-producing Lp90 seems to improve some rheological features.
Since millennia, humankind has exploited microbial diversity associated to give food matrices in ... more Since millennia, humankind has exploited microbial diversity associated to give food matrices in order to obtain fermented foods and beverages, resulting in products with improved quality and extended shelf life. This topic has received deserved and continuous interest in the scientific community, for the reason of its significance as a driver of innovation in the food and beverage sector. In this review paper, using sourdough as a model matrix, we provide some insights into the field, testifying the relevance as a transdisciplinary subject. Firstly, we encompassed the prokaryotic and eukaryotic microbial diversity associated with the sourdough ecosystems. The importance of this micro-biodiversity in the light of flour-related chemical diversity was examined. Finally, we highlighted the increasing interest in microbial-based applications oriented toward biocontrol solution in the field of sourdough-based products (i.e., bread).
In this study, we explored the diversity of yeasts and lactic acid bacteria (LAB) associated with... more In this study, we explored the diversity of yeasts and lactic acid bacteria (LAB) associated with six spontaneous sourdough fermentations from the northern part of the Apulian region (Italy). Bacterial and yeast isolates from sourdough were investigated by amplified ribosomal DNA restriction analysis (ARDRA) and restriction fragment length polymorphism (RFLP) analysis, respectively. The identification of the isolates was confirmed by sequencing bacterial 16S gene and yeast ITS1–5.8S–ITS2 rRNA gene amplicons. Microbiological analysis of all sourdough samples revealed that LAB and yeast counts ranged between 1.7 × 10⁵ and 6.5 × 10⁸ cfu/g, and 7.7 × 10⁵ and 2.5 × 10⁷ cfu/g, respectively. The molecular identification at species level revealed the occurrence of Lactobacillus plantarum as the dominant LAB and Saccharomyces cerevisiae as the dominant yeast species in all different sourdough samples. Then, the ability of all isolated strains to inhibit and/or reduce the growth of several se...
Abstract The dietary consumption of probiotics in the form of pharmaceuticals or functional food ... more Abstract The dietary consumption of probiotics in the form of pharmaceuticals or functional food can improve human health and contribute to disease prevention. However, the biological activity and health potential of food-delivered probiotics can be severely compromised by the stress conditions encountered by the microorganisms throughout the manufacture process, from probiotic preparation to their inclusion into food, subsequent storage and ingestion. Here, we give an account of the stress factors that can have major negative impacts on probiotic viability and functionality, with a focus on food-related environmental adverse conditions. We also describe some of the mechanisms elicited by the microbial cells to counteract these stresses and summarize a few relevant approaches proposed in literature to develop more robust and competitive probiotics by enhancing their stress tolerance, with the aim to improve the efficacy and health value of probiotic functional food.
Small heat shock proteins (sHSPs) are ubiquitous, low molecular weight (MW) proteins that share a... more Small heat shock proteins (sHSPs) are ubiquitous, low molecular weight (MW) proteins that share a conserved alpha-crystallin domain. sHSPs oligomers exhibit chaperon-like activities by interacting with unfolded substrates, thereby preventing their aggregation and precipitation. Unlike most lactobacilli, which have single shsp genes, three different sHSP-encoding genes, i.e., hsp1, hsp2, and hsp3, were previously identified in the probiotic Lactobacillus plantarum WCFS1. Early studies, including the characterization of the knock out (KO) mutant for hsp2, indicated a different organization and transcriptional regulation of these genes and suggested that the three L. plantarum sHSPs might accomplish different tasks in stress response. To unravel the role of sHSPs, KO mutants of hsp1 and hsp3 were generated using a Cre-lox based system. Mutation of either genes resulted in impaired growth capacity under normal conditions, heatstress and stresses typically found during host interactions and food technological process. However, survival to heat shock and the level of thermal stabilization of cytoplasmic proteins were similar between mutants and parental strain. Transcriptional analysis revealed that in the mutant genetic backgrounds there is an upregulated basal expression of the un-mutated mate hsps and other stress-related genes, which may compensate for the loss of HSP function, hence possibly accounting for the lack of a remarkable susceptibility to heat challenge. HSP3 seemed relevant for the induction of thermotolerance, while HSP1 was required for improved cryotolerance. Cell surface properties and plasma membrane fluidity were investigated to ascertain the possible membrane association of sHSP. Intriguingly, the loss of hsp1 was associated to a lower level of maximal membrane fluidity upon heat stress. A role for HSP1 in controlling and improving membrane fluidity is suggested which may pertains its cryoprotective function.
Lactic acid bacteria (LAB), a heterogeneous group of bacteria that produce lactic acid as the mai... more Lactic acid bacteria (LAB), a heterogeneous group of bacteria that produce lactic acid as the main product of carbohydrate degradation, play an important role in the production and protection of fermented foods. Moreover, beside the technological use of these microorganisms added to control and steer food fermentations, their beneficial healthy properties are largely overt. Thus, numerous LAB strains have obtained the probiotic status, which entails the ability to maintain and promote a good health of consumers. In particular, increasing consideration is being focused on probiotic microorganisms that can improve the human immune response against dangerous viral and fungal enemies. For such beneficial microbes, the term “immunobiotics” has been coined. Together with an indirect host-mediated adverse effect against undesirable microorganisms, also a direct antagonistic activity of several LAB strains has been largely demonstrated. The purpose of this review is to provide a fullest possible overview of the antiviral and antifungal activities ascribed to probiotic LAB. The interest in this research field is substantiated by a large number of studies exploring the potential application of these beneficial microorganisms both as biopreservatives and immune-enhancers, aiming to reduce and/or eliminate the use of chemical agents to prevent the development of pathogenic, infectious, and/or degrading causes.
More understanding of the risk-benefit effect of the glycoalkaloid tomatine is required to be abl... more More understanding of the risk-benefit effect of the glycoalkaloid tomatine is required to be able to estimate the role it might play in our diet. In this work, we focused on effects towards intestinal epithelial cells based on a Caco-2 model in order to analyze the influence on the cell monolayer integrity and on the expression levels of genes involved in cholesterol/sterol biosynthesis (LDLR), lipid metabolism (NR2F2), glucose and amino acid uptake (SGLT1, PAT1), cell cycle (PCNA, CDKN1A), apoptosis (CASP-3, BMF, KLF6), tight junctions (CLDN4, OCLN2) and cytokine-mediated signaling (IL-8, IL1β, TSLP, TNF-α). Furthermore, since the bioactivity of the compound might vary in the presence of a food matrix and following digestion, the influence of both pure tomatine and in vitro digested tomatine with and without tomato fruit matrix was studied. The obtained results suggested that concentrations <20 µg/mL of tomatine, either undigested or in vitro digested, do not compromise the via...
The enterovirus Coxsackievirus B4 (CV-B4) can infect different human tissues and provoke abnormal... more The enterovirus Coxsackievirus B4 (CV-B4) can infect different human tissues and provoke abnormal function or destruction of various organs and cells. Moreover, its infections have been linked to the onset of type 1 diabetes. Coxsackievirus B4 is classified as a "challenging virus", due to the intense yet vain efforts to find effective prevention and therapeutic agents, especially within biological compounds. Lactobacillus plantarum is a lactic acid bacterium that is endowed with probiotic properties, and holds great potential for applications in medical and food industry sectors. Several compounds produced by this microorganism have been associated with various benefits including antimicrobial activity. In this work, we investigated the possible antiviral abilities of two Lb. plantarum strains and their derivatives against CV-B4. The different assays carried out (e.g. pre-incubation, competition and post-infection, using HEp-2 cells as human cell model) suggest that the t...
Probiotics and antimicrobial proteins, Jan 19, 2018
Lactobacilli naturally present in the neonatal gut are believed to be beneficial for the human ho... more Lactobacilli naturally present in the neonatal gut are believed to be beneficial for the human hosts and are investigated as potential probiotics. In this study, we aimed to characterize six Lactobacillus plantarum strains derived from the feces of a breast-fed infant, for the development of new probiotic cultures. Our attention was focused on L. plantarum in reason of the presence, within such species, of both pro-technological and probiotic strains, i.e., a combination of particular interest to design tailored probiotic starter cultures for developing countries. The bacterial isolates exhibiting lactobacilli-like phenotypic characteristics were identified as members of the L. plantarum group by 16S rRNA gene sequencing, and their diversity was evaluated by randomly amplified polymorphic DNA (RAPD) PCR patterns. The selected strains were screened for probiotic potential through in vitro tests. Firstly, bacterial survival was evaluated in an in vitro system simulating the human oro-...
Lactic acid bacteria (LAB) are a heterogeneous group of Gram-positive bacteria that comprise seve... more Lactic acid bacteria (LAB) are a heterogeneous group of Gram-positive bacteria that comprise several species which have evolved in close association with humans (food and lifestyle). While their use to ferment food dates back to very ancient times, in the last decades, LAB have attracted much attention for their documented beneficial properties and for potential biomedical applications. Some LAB are commensal that colonize, stably or transiently, host mucosal surfaces, inlcuding the gut, where they may contribute to host health. In this review, we present and discuss the main factors enabling LAB adaptation to such lifestyle, including the gene reprogramming accompanying gut colonization, the specific bacterial components involved in adhesion and interaction with host, and how the gut niche has shaped the genome of intestine-adapted species. Moreover, the capacity of LAB to colonize abiotic surfaces by forming structured communities, i.e., biofilms, is briefly discussed, taking into account the main bacterial and environmental factors involved, particularly in relation to food-related environments. The vast spread of LAB surface-associated communities and the ability to control their occurrence hold great potentials for human health and food safety biotechnologies.
The sparkling wine market has expanded in recent years, boosted by the increasing demand of the g... more The sparkling wine market has expanded in recent years, boosted by the increasing demand of the global market. As for other fermented beverages, technological yeasts and bacteria selected to design commercial starter cultures represent key levers to maximize product quality and safety. The increasing economic interest in the sector of sparkling wine has also implied a renewed interest in microbial resource management. In this review, after a brief introduction, we report an overview of the main characterization criteria in order to select Saccharomyces cerevisiae strains suitable for use as starter cultures for the production of base wines and to drive re-fermentation of base wines to obtain sparkling wines. Particular attention has been reserved to the technological characterization aspects of re-fermenting phenotypes. We also analysed the possible uses of selected non-Saccharomyces and malolactic strains in order to differentiate specific productions. Finally, we highlighted the main safety aspects related to microbes of enological interest and underlined some microbial-based biotechnological applications helpful to pursue product and process innovations. Overall, the sparkling wine industry may find a relevant benefit from the exploitation of the wide resources associated with vineyard/wine microbial diversity.
Abstract Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as pr... more Abstract Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as proteins, carbohydrates, lipids, vitamins, and minerals. These compounds comprise an ideal growth substrate for both harmful and beneficial microorganisms, making milk a perishable raw material. As a result, milk contains numerous microorganisms that originate either from the animal itself, even when the animal is clinically healthy, or from the environment during the collection and storage of milk. The increasing social, economic, and research on non-cow types of milk and its products illustrates the renewed interest in the microbiota associated with these matrices. The present work is an overview of existing knowledge on the microbiota of sheep, goat, buffalo, camel, equine, yak, and human milk and their products. Relevant studies concerning both classical microbiological approaches as well as advanced molecular methods are included.
Lactobacillus plantarum is one of the most versatile species extensively used in the food industr... more Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect. This study emphasizes the tempting use of the tested L. plantarum strains and/or their CFS as antimicrobial agents against food-borne pathogens.
The influence of oat and barley β-glucans on some features of diverse probiotic strains was inves... more The influence of oat and barley β-glucans on some features of diverse probiotic strains was investigated. Neither barley nor oat β-glucans supported the growth of any of the investigated strains when used as the sole carbon source in fermentation media. No protective effect by β-glucans was observed on bacterial survival to in vitro simulation of the human digestive tract. Moreover, the presence of β-glucans did not enhance the ability of probiotic bacteria to adhere on human intestinal cells. The immunomodulatory activities of probiotics, β-glucans, and their combinations were studied and compared in vitro by transcriptional analysis of immune-related genes on LPS-stimulated THP-1 macrophages. Immune modulating properties were evidenced. In particular, mixtures of probiotic microorganisms and barley β-glucans exhibited synergistic effects in modulating the transcriptional level of several immune-related genes, leading to an overall enhanced anti-inflammatory effect. Together, these findings suggest a promising application of probiotic bacteria and β-glucans in the preparation of dietary mixtures presenting health-promoting features such as immunomodulatory properties.
Abstract: Small heat shock proteins (sHsps) are ubiquitous conserved chaperone-like proteins invo... more Abstract: Small heat shock proteins (sHsps) are ubiquitous conserved chaperone-like proteins involved in cellular proteins protection under stressful conditions. In this study, a reverse transcription quantitative PCR (RT-qPCR) procedure was developed and used to quantify the transcript level of a small heat shock gene (shs) in the probiotic bacterium Lactobacillus acidophilus NCFM, under stress conditions such as heat (45 °C and 53 °C), bile (0.3 % w/v), hyperosmosis (1 M and 2.5 M NaCl), and low pH value (pH 4). The shs gene of L. acidophilus NCFM was induced by salt, high temperature and acidic stress, while repression was observed upon bile stress. Analysis of the 5 ' noncoding region of the hsp16 gene reveals the presence of an inverted repeat (IR) sequence (TTAGCACTC-N9-GAGTGCTAA) homologue to the controlling IR of chaperone expression (CIRCE) elements found in the upstream regulatory region of Gram-positive heat shock operons, suggesting that the hsp16 gene of L. acidoph...
Abstract: Currently, the majority of prebiotics in the market are derived from non-digestible oli... more Abstract: Currently, the majority of prebiotics in the market are derived from non-digestible oligosaccharides. Very few studies have focused on non-digestible long chain complex polysaccharides in relation to their potential as novel prebiotics. Cereals β-glucans have been investigated for immune-modulating properties and beneficial effects on obesity, cardiovascular diseases, diabetes, and cholesterol levels. Moreover, β-glucans have been reported to be highly fermentable by the intestinal microbiota in the caecum and colon, and can enhance both growth rate and lactic acid production of microbes isolated from the human intestine. In this work, we report the effects of food matrices containing barley β-glucans on growth and probiotic features of four Lactobacillus strains. Such matrices were able to improve the growth rate of the tested bacteria both in unstressed conditions and, importantly, after exposure to in vitro simulation of the digestive tract. Moreover, the effect of β-gl...
Fermented oat-based foods offer attractive prospects within the market of non-dairy functional pr... more Fermented oat-based foods offer attractive prospects within the market of non-dairy functional products, since they are suitable substrates for the delivery of probiotic microorganisms, and are significant sources of dietary fiber, both insoluble and soluble such as β-glucan, good quality fat and other phytochemicals important for human health In the present work, whole oat flour was fermented with probiotic Lactobacillus plantarum strains to produce new functional foods with improved nutritional and technological features. Viability of the probiotic and the main technological, physico-chemical, nutritional and sensorial parameters were monitored at 7, 14 and 21 days of cold storage. The microbial survival was higher than 5x10 8 cfu g-1 at the end of the shelf life. After the fermentation step, viscosity was higher in products inoculated with the exopolysaccharide-producing L. plantarum strain Lp90. However, a subsequent viscosity reduction was detected in all the samples throughout the storage period, consistent with the observed concentration decrease of the oat β-glucan. Vitamin B2 content was about twofold higher in products fermented by the riboflavin-overproducing LpB2, and in these samples the riboflavin concentration further increased during cold storage. Highlights Fermented oat-based foods are suitable substrates for delivery probiotic LAB. Riboflavin-overproducing LpB2 increased the vitamin B2 content of oat-based products. EPS-producing Lp90 seems to improve some rheological features.
Since millennia, humankind has exploited microbial diversity associated to give food matrices in ... more Since millennia, humankind has exploited microbial diversity associated to give food matrices in order to obtain fermented foods and beverages, resulting in products with improved quality and extended shelf life. This topic has received deserved and continuous interest in the scientific community, for the reason of its significance as a driver of innovation in the food and beverage sector. In this review paper, using sourdough as a model matrix, we provide some insights into the field, testifying the relevance as a transdisciplinary subject. Firstly, we encompassed the prokaryotic and eukaryotic microbial diversity associated with the sourdough ecosystems. The importance of this micro-biodiversity in the light of flour-related chemical diversity was examined. Finally, we highlighted the increasing interest in microbial-based applications oriented toward biocontrol solution in the field of sourdough-based products (i.e., bread).
In this study, we explored the diversity of yeasts and lactic acid bacteria (LAB) associated with... more In this study, we explored the diversity of yeasts and lactic acid bacteria (LAB) associated with six spontaneous sourdough fermentations from the northern part of the Apulian region (Italy). Bacterial and yeast isolates from sourdough were investigated by amplified ribosomal DNA restriction analysis (ARDRA) and restriction fragment length polymorphism (RFLP) analysis, respectively. The identification of the isolates was confirmed by sequencing bacterial 16S gene and yeast ITS1–5.8S–ITS2 rRNA gene amplicons. Microbiological analysis of all sourdough samples revealed that LAB and yeast counts ranged between 1.7 × 10⁵ and 6.5 × 10⁸ cfu/g, and 7.7 × 10⁵ and 2.5 × 10⁷ cfu/g, respectively. The molecular identification at species level revealed the occurrence of Lactobacillus plantarum as the dominant LAB and Saccharomyces cerevisiae as the dominant yeast species in all different sourdough samples. Then, the ability of all isolated strains to inhibit and/or reduce the growth of several se...
Abstract The dietary consumption of probiotics in the form of pharmaceuticals or functional food ... more Abstract The dietary consumption of probiotics in the form of pharmaceuticals or functional food can improve human health and contribute to disease prevention. However, the biological activity and health potential of food-delivered probiotics can be severely compromised by the stress conditions encountered by the microorganisms throughout the manufacture process, from probiotic preparation to their inclusion into food, subsequent storage and ingestion. Here, we give an account of the stress factors that can have major negative impacts on probiotic viability and functionality, with a focus on food-related environmental adverse conditions. We also describe some of the mechanisms elicited by the microbial cells to counteract these stresses and summarize a few relevant approaches proposed in literature to develop more robust and competitive probiotics by enhancing their stress tolerance, with the aim to improve the efficacy and health value of probiotic functional food.
Small heat shock proteins (sHSPs) are ubiquitous, low molecular weight (MW) proteins that share a... more Small heat shock proteins (sHSPs) are ubiquitous, low molecular weight (MW) proteins that share a conserved alpha-crystallin domain. sHSPs oligomers exhibit chaperon-like activities by interacting with unfolded substrates, thereby preventing their aggregation and precipitation. Unlike most lactobacilli, which have single shsp genes, three different sHSP-encoding genes, i.e., hsp1, hsp2, and hsp3, were previously identified in the probiotic Lactobacillus plantarum WCFS1. Early studies, including the characterization of the knock out (KO) mutant for hsp2, indicated a different organization and transcriptional regulation of these genes and suggested that the three L. plantarum sHSPs might accomplish different tasks in stress response. To unravel the role of sHSPs, KO mutants of hsp1 and hsp3 were generated using a Cre-lox based system. Mutation of either genes resulted in impaired growth capacity under normal conditions, heatstress and stresses typically found during host interactions and food technological process. However, survival to heat shock and the level of thermal stabilization of cytoplasmic proteins were similar between mutants and parental strain. Transcriptional analysis revealed that in the mutant genetic backgrounds there is an upregulated basal expression of the un-mutated mate hsps and other stress-related genes, which may compensate for the loss of HSP function, hence possibly accounting for the lack of a remarkable susceptibility to heat challenge. HSP3 seemed relevant for the induction of thermotolerance, while HSP1 was required for improved cryotolerance. Cell surface properties and plasma membrane fluidity were investigated to ascertain the possible membrane association of sHSP. Intriguingly, the loss of hsp1 was associated to a lower level of maximal membrane fluidity upon heat stress. A role for HSP1 in controlling and improving membrane fluidity is suggested which may pertains its cryoprotective function.
Lactic acid bacteria (LAB), a heterogeneous group of bacteria that produce lactic acid as the mai... more Lactic acid bacteria (LAB), a heterogeneous group of bacteria that produce lactic acid as the main product of carbohydrate degradation, play an important role in the production and protection of fermented foods. Moreover, beside the technological use of these microorganisms added to control and steer food fermentations, their beneficial healthy properties are largely overt. Thus, numerous LAB strains have obtained the probiotic status, which entails the ability to maintain and promote a good health of consumers. In particular, increasing consideration is being focused on probiotic microorganisms that can improve the human immune response against dangerous viral and fungal enemies. For such beneficial microbes, the term “immunobiotics” has been coined. Together with an indirect host-mediated adverse effect against undesirable microorganisms, also a direct antagonistic activity of several LAB strains has been largely demonstrated. The purpose of this review is to provide a fullest possible overview of the antiviral and antifungal activities ascribed to probiotic LAB. The interest in this research field is substantiated by a large number of studies exploring the potential application of these beneficial microorganisms both as biopreservatives and immune-enhancers, aiming to reduce and/or eliminate the use of chemical agents to prevent the development of pathogenic, infectious, and/or degrading causes.
More understanding of the risk-benefit effect of the glycoalkaloid tomatine is required to be abl... more More understanding of the risk-benefit effect of the glycoalkaloid tomatine is required to be able to estimate the role it might play in our diet. In this work, we focused on effects towards intestinal epithelial cells based on a Caco-2 model in order to analyze the influence on the cell monolayer integrity and on the expression levels of genes involved in cholesterol/sterol biosynthesis (LDLR), lipid metabolism (NR2F2), glucose and amino acid uptake (SGLT1, PAT1), cell cycle (PCNA, CDKN1A), apoptosis (CASP-3, BMF, KLF6), tight junctions (CLDN4, OCLN2) and cytokine-mediated signaling (IL-8, IL1β, TSLP, TNF-α). Furthermore, since the bioactivity of the compound might vary in the presence of a food matrix and following digestion, the influence of both pure tomatine and in vitro digested tomatine with and without tomato fruit matrix was studied. The obtained results suggested that concentrations <20 µg/mL of tomatine, either undigested or in vitro digested, do not compromise the via...
The enterovirus Coxsackievirus B4 (CV-B4) can infect different human tissues and provoke abnormal... more The enterovirus Coxsackievirus B4 (CV-B4) can infect different human tissues and provoke abnormal function or destruction of various organs and cells. Moreover, its infections have been linked to the onset of type 1 diabetes. Coxsackievirus B4 is classified as a "challenging virus", due to the intense yet vain efforts to find effective prevention and therapeutic agents, especially within biological compounds. Lactobacillus plantarum is a lactic acid bacterium that is endowed with probiotic properties, and holds great potential for applications in medical and food industry sectors. Several compounds produced by this microorganism have been associated with various benefits including antimicrobial activity. In this work, we investigated the possible antiviral abilities of two Lb. plantarum strains and their derivatives against CV-B4. The different assays carried out (e.g. pre-incubation, competition and post-infection, using HEp-2 cells as human cell model) suggest that the t...
Probiotics and antimicrobial proteins, Jan 19, 2018
Lactobacilli naturally present in the neonatal gut are believed to be beneficial for the human ho... more Lactobacilli naturally present in the neonatal gut are believed to be beneficial for the human hosts and are investigated as potential probiotics. In this study, we aimed to characterize six Lactobacillus plantarum strains derived from the feces of a breast-fed infant, for the development of new probiotic cultures. Our attention was focused on L. plantarum in reason of the presence, within such species, of both pro-technological and probiotic strains, i.e., a combination of particular interest to design tailored probiotic starter cultures for developing countries. The bacterial isolates exhibiting lactobacilli-like phenotypic characteristics were identified as members of the L. plantarum group by 16S rRNA gene sequencing, and their diversity was evaluated by randomly amplified polymorphic DNA (RAPD) PCR patterns. The selected strains were screened for probiotic potential through in vitro tests. Firstly, bacterial survival was evaluated in an in vitro system simulating the human oro-...
Lactic acid bacteria (LAB) are a heterogeneous group of Gram-positive bacteria that comprise seve... more Lactic acid bacteria (LAB) are a heterogeneous group of Gram-positive bacteria that comprise several species which have evolved in close association with humans (food and lifestyle). While their use to ferment food dates back to very ancient times, in the last decades, LAB have attracted much attention for their documented beneficial properties and for potential biomedical applications. Some LAB are commensal that colonize, stably or transiently, host mucosal surfaces, inlcuding the gut, where they may contribute to host health. In this review, we present and discuss the main factors enabling LAB adaptation to such lifestyle, including the gene reprogramming accompanying gut colonization, the specific bacterial components involved in adhesion and interaction with host, and how the gut niche has shaped the genome of intestine-adapted species. Moreover, the capacity of LAB to colonize abiotic surfaces by forming structured communities, i.e., biofilms, is briefly discussed, taking into account the main bacterial and environmental factors involved, particularly in relation to food-related environments. The vast spread of LAB surface-associated communities and the ability to control their occurrence hold great potentials for human health and food safety biotechnologies.
The sparkling wine market has expanded in recent years, boosted by the increasing demand of the g... more The sparkling wine market has expanded in recent years, boosted by the increasing demand of the global market. As for other fermented beverages, technological yeasts and bacteria selected to design commercial starter cultures represent key levers to maximize product quality and safety. The increasing economic interest in the sector of sparkling wine has also implied a renewed interest in microbial resource management. In this review, after a brief introduction, we report an overview of the main characterization criteria in order to select Saccharomyces cerevisiae strains suitable for use as starter cultures for the production of base wines and to drive re-fermentation of base wines to obtain sparkling wines. Particular attention has been reserved to the technological characterization aspects of re-fermenting phenotypes. We also analysed the possible uses of selected non-Saccharomyces and malolactic strains in order to differentiate specific productions. Finally, we highlighted the main safety aspects related to microbes of enological interest and underlined some microbial-based biotechnological applications helpful to pursue product and process innovations. Overall, the sparkling wine industry may find a relevant benefit from the exploitation of the wide resources associated with vineyard/wine microbial diversity.
Abstract Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as pr... more Abstract Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as proteins, carbohydrates, lipids, vitamins, and minerals. These compounds comprise an ideal growth substrate for both harmful and beneficial microorganisms, making milk a perishable raw material. As a result, milk contains numerous microorganisms that originate either from the animal itself, even when the animal is clinically healthy, or from the environment during the collection and storage of milk. The increasing social, economic, and research on non-cow types of milk and its products illustrates the renewed interest in the microbiota associated with these matrices. The present work is an overview of existing knowledge on the microbiota of sheep, goat, buffalo, camel, equine, yak, and human milk and their products. Relevant studies concerning both classical microbiological approaches as well as advanced molecular methods are included.
Lactobacillus plantarum is one of the most versatile species extensively used in the food industr... more Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect. This study emphasizes the tempting use of the tested L. plantarum strains and/or their CFS as antimicrobial agents against food-borne pathogens.
The influence of oat and barley β-glucans on some features of diverse probiotic strains was inves... more The influence of oat and barley β-glucans on some features of diverse probiotic strains was investigated. Neither barley nor oat β-glucans supported the growth of any of the investigated strains when used as the sole carbon source in fermentation media. No protective effect by β-glucans was observed on bacterial survival to in vitro simulation of the human digestive tract. Moreover, the presence of β-glucans did not enhance the ability of probiotic bacteria to adhere on human intestinal cells. The immunomodulatory activities of probiotics, β-glucans, and their combinations were studied and compared in vitro by transcriptional analysis of immune-related genes on LPS-stimulated THP-1 macrophages. Immune modulating properties were evidenced. In particular, mixtures of probiotic microorganisms and barley β-glucans exhibited synergistic effects in modulating the transcriptional level of several immune-related genes, leading to an overall enhanced anti-inflammatory effect. Together, these findings suggest a promising application of probiotic bacteria and β-glucans in the preparation of dietary mixtures presenting health-promoting features such as immunomodulatory properties.
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Papers by Mattia Arena