Fluorescence in situ hybridisation detection of Lactobacillus plantarum group on olives to be used in natural fermentations

At present there are very few studies on the bacterial diversity of olives and on the importance of the microbial species for the fermentation of olives aimed to table olives production. Most of the authors report on the occurrence of Lactobacillus plantarum as principal member of these communities or at least as the species responsible for the fermentation. In this study, fluorescence in situ hybridisation (FISH) with 16S rRNA probes was used to evaluate the occurrence of L. plantarum in olives. A 18-bp oligonucleotide probe was used in FISH experiments to evaluate the specificity of detection among Lactobacillus species. The probe was tested against 30 Lactobacillus species and appeared to be specific for L. plantarum, L. paraplantarum and L. pentosus. The probe was then used to investigate the occurrence of these species in 25 samples of olives (cultivar “Leccino”) collected in Campania region (Southern Italy). The olives were washed in a saline solution and the suspensions were then analysed by FISH and observed by fluorescence microscopy. No hybridisation signal was detected in at least 30 fields of observation when the L. plantarum-specific probe was used, probably due to the low sensitivity of the FISH method. Olive samples were plated on Rogosa agar and about 40% of the samples did not give growth after 5 days. When colony growth was observed, bulk cells from Rogosa agar plates were collected and analysed by DNA extraction followed by 16S rDNA Polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). The different microbial species were identified by direct sequencing of DGGE bands. Leuconostoc pseudomesenteroides was the most frequently found species, occurring in more than 50% of the samples that had shown growth on Rogosa agar. The closest relatives of the species of the genera: Leuconostoc, Pediococcus, Pseudomonas and Raoultella were also identified suggesting that guided fermentation by using selected LAB starters is advisable for a safe and desired table olives production

Technological activities of Staphylococcus carnosus and Staphylococcus simulans strains isolated from fermented sausages

The aim of this study was to determine the technological properties of 2 strains of Staphylococcus simulans (Ssm12, Ssm21) and 4 strains of S. carnosus (SC28, SC31, SC54 and SC55) for the selection of a potential starter cultures to employ in the processing of dry fermented sausages. The strains were studied to evaluate nitrate reductase, proteolytic, lipolytic, decarboxylase and antioxidant activities as well as growth ability at different temperatures, pH and NaCl concentrations. Nitrate reductase activity was determined at 15, 20 and 30 degrees C. By spectrophotometric method all the strains were able to reduce nitrate to nitrite at the different temperatures but these results were not confirmed by the agar plate method. Antioxidant and lipolytic activities were evaluated by spectrophotometric assay. All the strains showed antioxidative enzymes superoxide dismutase (SOD) and catalase whereas all appeared unable to hydrolyse pork fat. Proteolytic activity was determined by agar plate method, spectrophotometric assay (OPA) and sodium dodecyl sulphate gel-electrophoresis (SDS-PAGE) and all strains appeared to be able to hydrolyse sarcoplasmic proteins but not myofibrillar proteins. Finally, all the strains grew at 15 and 20 degrees C, in presence of 10%, 15% and 20% of NaCl and at pH 5.0 and 5.5 and were unable to produce histamine, cadaverine and putrescine. The results showed that all strains studied possess useful technological activities that would make them eligible as a good starter cultures for fermented sausages

Adaptation to aerobic environment of Lactobacillus johnsonii/gasseri strains

Oxygen is considered one of the main factors affecting probiotic bacteria survival due to the induction of oxidative damages caused by the action of reactive oxygen species (ROS). It has been shown that oxidative stress resistance in lactic acid bacteria is strongly dependent on the type of cell metabolism. Shift from fermentative to respiratory metabolism (through the addition of heme and menaquinone and in presence of oxygen) was associated to increase in biomass, long-term survival, and production of antioxidant enzymes. The aim of this work was to investigate the effect of aerobic (presence of oxygen) and respiratory (presence of oxygen, heme, and menaquinone) cultivation on the growth kinetic, catalase production, oxygen uptake, and oxidative stress response of Lactobacillus johnsonii/gasseri strains previously isolated from infant feces. Seven strains showed to consume oxygen under aerobic and respiratory conditions. The strain AL5 showed a catalase activity in both growth conditions, while AL3 showed this activity only in respiratory condition. Respiratory condition improved their tolerance to oxidative compounds (hydrogen peroxide and ROS generators) and further they showed promising probiotic features. The exploration of respiratory competent phenotypes with probiotic features may be extremely useful for the development of competitive starter or probiotic cultures

Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics.

A rapid screening method was used to isolate potentially probiotic Lactobacillus strains from fermented sausages after enrichment in MRS broth at pH 2.5 followed by bile salt stressing (1% bile salts w/v). One hundred and fifty acid- and bile-resistant strains were selected, avoiding preliminary and time-consuming isolation steps. Strains were further characterized for survival at pH 2.5 for 3 h in phosphate-buffered saline and for growth in the presence of 0.3% bile salts with and without pre-exposure at low pH. Twentyeight strains showed a survival >80% at pH 2.5 for 3 h; moreover, most of the strains were able to grow in the presence of 0.3% bile salts. Low pH and bile resistance was shown to be dependent on both the species, identified by phenotypic and molecular methods, and the strain tested. This is the first report on the direct selection of potentially probiotic lactobacilli from dry fermented sausages. Technologically interesting strains may be used in the future as probiotic starter cultures for novel fermented sausage manufacture

Layer-by-Layer Coating of Single-Cell Lacticaseibacillus rhamnosus to Increase Viability Under Simulated Gastrointestinal Conditions and Use in Film Formation

Probiotics and prebiotics are widely used as functional food ingredients. Viability of probiotics in the food matrix and further in the digestive system is still a challenge for the food industry. Different approaches were used to enhance the viability of probiotics including microencapsulation and layer-by-layer cell coating. The of aim of this study was to evaluate the viability of coated Lacticaseibacillus rhamnosus using a layer-by-layer (LbL) technique with black seed protein (BSP) extracted from Nigella sativa defatted seeds cakes (NsDSC), as a coating material, with alginate, inulin, or glucomannan, separately, and the final number of coating layers was 3. The viable cell counts of the plain and coated L. rhamnosus were determined under sequential simulated gastric fluid (SGF) for 120 min and simulated intestinal fluid (SIF) for 180 min. Additionally, the viability after exposure to 37, 45, and 55 C for 30 min was also determined. Generally, the survivability of coated L. rhamnosus showed significant (p   0.05) improvement (<4, 3, and 1.5 logs reduction for glucomannan, alginate and inulin, respectively) compared with plain cells ( 6.7 log reduction) under sequential exposure to SGF and SIF. Moreover, the cells coated with BSP and inulin showed the best protection for L. rhamnosus under high temperatures. Edible films prepared with pectin with LbL-coated cells showed significantly higher values in their tensile strength (TS) of 50% and elongation at the break (EB) of 32.5% than pectin without LbL-coated cells. The LbL technique showed a significant protection of probiotic cells and potential use in food application

Application of ultrasound and microencapsulation on Limosilactobacillus reuteri DSM 17938 as a metabolic attenuation strategy for tomato juice probiotication

Counteracting probiotic-induced physicochemical and sensory changes is a challenge in the development of probiotic beverages. The aim of the study is to apply ultrasound and microencapsulation for the attenuation of Limosilactobacillus reuteri DSM 17938 to avoid change in a probiotic tomato juice. Preliminarily, six ultrasound treatments were applied. Probiotic survival in acid environment (pH 2.5) and bile salts (1.5 g/l) after ultrasound treatment was also studied. The probiotic was inoculated in tomato juice in four forms: free cells (PRO-TJ), sonicated-free cells (US-TJ), untreated-microencapsulated (PRO-MC-TJ) and sonicated-microencapsulated cells (US-MC-TJ). Probiotic viability and pH were monitored during 28 days of storage at 4 and 20 °C. Sensory analysis was performed for PRO-TJ and US-MC-TJ sample (4 °C). Ultrasound (57 W for 6 min) did not affect cell survival and transitorily modulated probiotic acidifying capacity; it reduced probiotic survival in acidic environment but increased probiotic survival in bile salts solution. Ultrasound was effective in maintain pH value of tomato juice but only at 4 °C. Instead, microencapsulation with sodium-alginate leads to a more stable probiotic juice, particularly at 20 °C. Finally, probiotication slightly modified some sensory attributes of the juice. This study shows the potential of ultrasound and microencapsulation as attenuation strategies and highlights the need for process optimization to increase ultrasound efficacy

Acid hydrolysis of spent coffee grounds: effects on possible prebiotic activity of oligosaccharides

Abstract Background Spent coffee grounds (SCG) are a promising source of natural by-products which can be used for different purposes. In this work, a possible use of oligosaccharides isolated from SCG as functional ingredients was investigated. SCGs were treated with an acid hydrolysis at high temperature (200 °C) in a closed reactor setting reaction time of 30, 60 and 90 s depending on the sample (original or defatted). A comprehensive study of the resulted water-soluble hydrolysate using a high-resolution mass spectrometry analysis was performed. Additionally, the growth of four Lactobacillus strains was tested to assess the prebiotic potential of the hydrolysate. Results Oligosaccharide chains formed by hexoses with a degree of polymerization ranging from 3 to 6 were identified and characterized. Regardless of the composition and the reaction time of hydrolysis, the bacterial activity of SCG extracts exhibited significantly higher values than the well-known versatile carbohydrate used by food industry, i.e., inulin. Conclusions The results pave the way toward the use of hydrolysate SCG as an innovative ingredient intended to fortify food formulations. The diversity in coffee oligosaccharides composition suggests their selective prebiotic activity for specific bacterial strains. Graphical Abstrac

Gut microbiome and blood glucose control in type 1 diabetes: a systematic review

ObjectiveThe risk of developing micro- and macrovascular complications is higher for individuals with type 1 diabetes (T1D). Numerous studies have indicated variations in gut microbial composition between healthy individuals and those with T1D. These changes in the gut ecosystem may lead to inflammation, modifications in intestinal permeability, and alterations in metabolites. Such effects can collectively impact the metabolic regulation system, thereby influencing blood glucose control. This review aims to explore the relationship between the gut microbiome, inflammation, and blood glucose parameters in patients with T1D.MethodsGoogle Scholar, PubMed, and Web of Science were systematically searched from 2003 to 2023 using the following keywords: “gut microbiota,” “gut microbiome,” “bacteria,” “T1D,” “type 1 diabetes,” “autoimmune diabetes,” “glycemic control,” “glucose control,” “HbA1c,” “inflammation,” “inflammatory,” and “cytokine.” The examination has shown 18,680 articles with relevant keywords. After the exclusion of irrelevant articles, seven observational papers showed a distinct gut microbial signature in T1D patients.ResultsThis review shows that, in T1D patients, HbA1c level was negatively correlated with abundance of Prevotella, Faecalibacterium, and Ruminococcaceae and positively correlated with abundance of Dorea formicigenerans, Bacteroidetes, Lactobacillales, and Bacteriodes. Instead, Bifidobacteria was negatively correlated with fasting blood glucose. In addition, there was a positive correlation between Clostridiaceae and time in range. Furthermore, a positive correlation between inflammatory parameters and gut dysbiosis was revealed in T1D patients.ConclusionWe draw the conclusion that the gut microbiome profiles of T1D patients and healthy controls differ. Patients with T1D may experience leaky gut, bacterial translocation, inflammation, and poor glucose management due to microbiome dysbiosis. Direct manipulation of the gut microbiome in humans and its effects on gut permeability and glycemic control, however, have not been thoroughly investigated. Future research should therefore thoroughly examine other potential pathophysiological mechanisms in larger studies