Viable Lactobacillus Research Articles

PSV-5 Characterizing the activity of bovine Lactobacillus strains against the top 7 Shiga-toxin Producing Escherichia coli strains

Abstract Shiga toxigenic Escherichia coli (STEC), specifically serogroups O26, O45, O111, O103, O121, O145, and O157 (Top 7), pose significant health risks. Cattle are the primary reservoir of STEC, and effective intervention remains elusive. The identification of Lactobacillus spp. with antimicrobial activity as direct-fed microbials (DFMs) presents a potential strategy for pathogen reduction. These strains are not only capable of favorably modulating the gut microbiota but also possess the innate ability to produce bacteriocins that can target specific pathogens including STEC. Selection of viable anti-STEC Lactobacillus strains requires assessment of the presence or absence of prophage that may revert from the lysogenic to lytic phase and affect their efficacy. This study aimed to characterize anti-STEC potential of 14 Lactobacillus strains isolated from cattle and to identify their associated bacteriocins and prophages. Antimicrobial activity of 14 Lactobacillus strains against the top 7 STEC strains was determined using live culture spot assays and cell-free supernatants (CFS) well-diffusion assays. To further assess their anti-STEC efficacy, Lactobacillus strains with the strongest activity were selected and further evaluated in MRS broth culture and rumen fluid. The ability of Lactobacillus strains to inhibit STEC attachment to Caco-2 epithelial cells was also assessed. Prokka 1.14.6, and Bagel5 were utilized to identify genes encoding bacteriocins. Additionally, PHASTEST 3.0 was utilized to identify prophages. All 14 strains demonstrated antimicrobial activity against the top 7 STEC serogroups. Notably, L. agilis strains L3 and L6, with genome sizes of 2,063,576 bp and 2,071,091 bp respectively, and L. amylovorus L7 (883,016 bp) exhibited the greatest anti-STEC activities, targeting O157, O26, O45, and O103 for L3 and L6, and O157, O45, and O103 for L7. The CFS from these strains showed anti-STEC activity at pH 4.4-4.6, but not at 7, suggesting that organic acids played a role in inhibition. In MRS, DFM reduced STEC O157:H7 by 5 to 7.4 logs after 24 h. However, in rumen fluid, no notable reduction in STEC O157:H7 was observed. Bioinformatic analysis revealed that the L7 genome encoded Helveticin-J, a Class III bacteriocin. Prophage identified included 1 intact (51,818 bp) and 1 non-intact (17,267 bp) in L3, and 2 intact (46,916 bp, 20,295 bp) and 1 non-intact (13,939 bp) in L6, while no prophage was identified in L7. This study identified novel DFMs with activity against STEC, but future studies with cattle are required to confirm their ability to inhibit STEC.

Sustained dual delivery of metronidazole and viable Lactobacillus crispatus from 3D-printed silicone shells

Bacterial vaginosis (BV) is an imbalance of the vaginal microbiome in which there are limited lactobacilli and an overgrowth of anaerobic and fastidious bacteria such as Gardnerella. The propensity for BV recurrence is high, and therapies involving multiple treatment modalities are emerging to meet this need. However, current treatments requiring frequent therapeutic administration are challenging for patients and impact user compliance. Three-dimensional (3D)-printing offers a novel alternative to customize platforms to facilitate sustained therapeutic delivery to the vaginal tract. This study designed a novel vehicle intended for dual sustained delivery of both antibiotic and probiotic. 3D-printed compartmental scaffolds consisting of an antibiotic-containing silicone shell and a core containing probiotic Lactobacillus were developed with multiple formulations including biomaterials sodium alginate (SA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyethylene oxide (PEO), and kappa-carrageenan (KC). The vehicles were loaded with 50 μg of metronidazole/mg polymer and 5 × 107 CFU of L. crispatus/mg scaffold. Metronidazole-containing shells exhibited cumulative drug release of 324.2 ± 31.2 μg/mL after 14 days. Multiple polymeric formulations for the probiotic core demonstrated cumulative L. crispatus recovery of >5 × 107 CFU/mg scaffold during this timeframe. L. crispatus-loaded polymeric formulations exhibited ≥2 log CFU/mL reduction in free Gardnerella in the presence of VK2/E6E7 vaginal epithelial cells. As a first step towards the goal of facilitating patient compliance, this study demonstrates in vitro effect of a novel 3D-printed dual antibiotic and probiotic delivery platform to target BV.

Enhanced viability of Lactobacillus spp. via encapsulation in hyaluronan/PVA hybrid electrospun composites for vaginal drug delivery

BackgroundVaginal dysbiosis, a change in the beneficial vaginal microbiome, leads to a significant depletion in the essential lactobacilli thus increasing the possibility of vaginal infections such as bacterial vaginosis. Probiotics have gained more attention as a means of delivering exogenous lactobacilli but one of the challenges in delivery strategies is maintaining and improving their viability. The objective of this study is to enhance the viability of Lactobacillus spp., via encapsulation in hyaluronic acid/polyvinyl alcohol hybrid electrospun nanofibers. Polyvinyl alcohol (PVA) and hyaluronic acid (HA) composite nanofibers integrated with Lactobacillus spp. were fabricated by electrospinning. The survival of Lactobacillus spp. after its immobilization in electrospun nanofibers with polyvinyl alcohol and hyaluronic acid was evaluated.ResultsScanning electron microscopy indicated larger average diameters in PVA/HA nanofibers with Lactobacillus spp. encapsulation (0.189 ± 0.041 µm to 0.231 ± 0.061 µm between D3 and D4, and 0.177 ± 0.043 µm to 0.212 ± 0.041 µm between D5 and D6) which showed that the nanofibers had the bacterial cells successfully enclosed in them. The viability of the lactic acid bacteria enclosed in the PVA/HA nanofibers was observed to decrease by more than 2-log units.ConclusionThe electrospun nanofiber-based delivery system is promising for the encapsulation and delivery of lactic acid bacteria to the vagina to combat recurrent vaginal infections such as bacterial vaginosis.

Comparative effects of viable Lactobacillus rhamnosus GG and its heat-inactivated paraprobiotic in the prevention of high-fat high-fructose diet-induced non-alcoholic fatty liver disease in rats.

Nonalcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver alterations worldwide, being gut microbiota dysbiosis one of the contributing factors to its development. The aim of this research is to compare the potential effects of a viable probiotic (Lactobacillus rhamnosus GG) with those exerted by its heat-inactivated paraprobiotic counterpart in a dietary rodent model of NAFLD. The probiotic administration effectively prevented the hepatic lipid accumulation induced by a high-fat high-fructose diet feeding, as demonstrated by chemical (lower TG content) and histological (lower steatosis grade and lobular inflammation) analyses. This effect was mainly mediated by the downregulation of lipid uptake (FATP2 protein expression) and upregulating liver TG release to bloodstream (MTTP activity) in rats receiving the probiotic. By contrast, the effect of the paraprobiotic preventing diet-induced liver lipid accumulation was milder, and mainly derived from the downregulation of hepatic de novo lipogenesis (SREBP-1c protein expression and FAS activity) and TG assembly (DGAT2 and AQP9 protein expression). The obtained results demonstrate that under these experimental conditions, the effects induced by the administration of viable L. rhamnosus GG preventing liver lipid accumulation in rats fed a diet rich in saturated fat and fructose differ from those induced by its heat-inactivated paraprobiotic counterpart.

O-268 Re-considering the endometrial microbiome: a comparison of DNA- and RNA-based sequencing techniques within the same cohort

Abstract Study question Does the analysis of the endometrial microbiome provide the same information when using DNA or RNA sequencing-based techniques? Summary answer DNA vs. RNA-based microbiome analysis techniques demonstrate significant microbial compositional differences, meaning that the previous endometrial 16S rRNA gene-based microbiome analysis results might be overestimated. What is known already In recent years, high-throughput sequencing technologies have revolutionised the field of reproductive microbiome research. Our understanding of the composition of endometrial microbiome predominantly relies on DNA-based 16S rRNA gene profiling method. Regardless of its effectiveness and low cost, its underestimation of microbial diversity, abundance, or functionality and lack of detection precision on species level have been highlighted. The meta-transcriptome analysis, RNA-based method, overcomes these limitations and identifies functional genes that are actively expressed by the alive microbes. This study aims to elucidate for the first time the endometrial microbiome using a dual approach of 16S rRNA gene-method and meta-transcriptomic analyses. Study design, size, duration In total forty-seven women with infertility at ages 27-42 years old were enrolled in the present study. Paired endometrial samples, endometrial brushing and Pipelle biopsy were collected from each participant at the mid-secretory menstrual phase (LH + 7/9). Participants/materials, setting, methods Endometrial samples were collected using Tao Brush for 16S rRNA analysis (DNA analysis) and Pipelle endometrial curette for meta-transcriptomic analysis (total RNA analysis). QIAamp UCP Pathogen Mini Kit was used for DNA extraction and 16S rRNA gene V3-V4 regions were sequenced on MiSeq. RNA was extracted using miRNeasy Micro kit, with rRNA removal by RiboZero kit and Libraries were generated using Stranded Total RNA Prep. Taxonomy was assigned by using Kraken2 (v2.2.1), Bracken (v2.7). Main results and the role of chance To the best of our knowledge, this is the first study to compare endometrial microbial identification using both metagenomics (16S rRNA sequencing) and meta-transcriptomics (meta-RNA sequencing) within the same cohort. Analysis of the composition of the microorganisms by bacterial 16S rRNA gene sequencing revealed that the most abundant bacterial genus in the endometrium was Lactobacillus. In the meta-transcriptome analysis, no microorganisms belonging to the Lactobacillus genus were detected in high abundance. This indicates that the relative abundance of this genus at the DNA level does not necessarily imply microbial activity or the presence of viable Lactobacillus in the endometrium. The meta-transcriptomics analysis detected microbial taxa such as Staphylococcus, Bacillus, Streptococcus, and Burkholderia as the most dominant. The results suggest that DNA-based detection of microorganisms in the endometrium may be overestimated due to the presence of naked DNA sequences or microorganisms from the vagina and cervix. Furthermore, RNA-based analysis demonstrates a population of active microorganisms different from the image shown by DNA analysis. Altogether, our study results indicate that the uterine microenvironment, as previously identified through DNA sequencing, may partly reflect the vaginocervical microenvironment, with the uterine tissue being a low biomass site not dominated by lactobacilli. Limitations, reasons for caution The endometrial samples were paired; however, DNA was analysed in the endometrial brushing sample while RNA in the tissue biopsy, which might result in some differences in microbial composition. Wider implications of the findings Contrary to the general belief of the Lactobacillus dominance in the human endometrium, our study suggests that the endometrial microenvironment may be harbouring DNA fragments and/or cells of lactobacilli originating from the lower reproductive tract. Our study results call out to re-consider/re-analyse the endometrial microbiome in health and disease. Trial registration number not applicable

Viability-PCR for the selective detection of Lactobacillus acidophilus and Bifidobacterium bifidum in live bacteria-containing products.

To exert their beneficial effects, microorganisms used in live bacteria-containing products must be viable and present in certain amounts. In this study, we developed a viability assay based on quantitative PCR coupled with propidium monoazide for the identification and enumeration of viable Lactobacillus acidophilus and Bifidobacterium bifidum. In order to optimize the protocol, the thermal inactivation conditions for the two target microorganisms and the PMA concentration inhibiting DNA amplification from the dead cells while allowing it from the live cells were first determined. The viability-PCR protocol was then applied to analyze a commercial product containing the two microorganisms. The quantities of both microorganisms determined using viability-PCR in the tested product were significantly higher than those obtained using the standard plate count, suggesting the presence of bacteria in a viable but non-culturable physiological state. Moreover, lower amounts of the two microorganisms were detected using viability-PCR compared to those achieved using quantitative PCR, possibly because of the presence of dead cells in the samples. Our results suggest that the viability-PCR method proposed here is a suitable alternative for rapid and accurate quantification and assessment of the viability of L. acidophilus and B. bifidum and could be easily adopted in the quality control screening of live bacteria-containing products.

Extraction, Characterisation and Evaluation of Antioxidant and Probiotic Growth Potential of Water-Soluble Polysaccharides from Ulva rigida Macroalgae.

Ulva rigida green macroalgae contain a variety of polysaccharides. A recent study investigated the optimum concentration and yield of polysaccharide extraction from oven-dried U. rigida biomass using a water-soluble polysaccharide extraction method that adhered to safety standards. This study utilised complete factorial experiments to examine the effects of varying factors on polysaccharide extraction. Results showed a positive correlation between increased levels of all factors and higher polysaccharide extraction yield. This study also found that the main factors and their interaction had a significant impact on the extracted polysaccharides from U. rigida. The highest polysaccharide content and yield were 9.5 mg/mL and 189 mg/g, respectively. Water-soluble polysaccharides demonstrated the presence of reducing sugar (8 mg/g), phenolics (0.69 mg/g) and flavonoids (1.42 mg/g) and exhibited antioxidant properties. Results revealed that freeze-dried polysaccharide powders were primarily composed of the monosaccharide rhamnose. Preliminary results on the effect of these powders on probiotics demonstrated that supplementation of polysaccharides from U. rigida promoted viable Lactobacillus rhamnosus ATCC 53103 growth during cultivation. This discovery has the potential to revolutionise the human food industry and promote the development of functional ingredients for novel and future food products, with numerous applications in the nutraceutical and pharmaceutical industries.

Análisis de microorganismos funcionales y su relación con parámetros fisicoquímicos del suelo en un bosque reservado

Este estudio aborda la necesidad de entender la presencia microorganismos funcionales en suelos tropicales, proporcionando información valiosa para la conservación y manejo sostenible. Tiene como principal objetivo analizar la variabilidad de microorganismos funcionales en diferentes profundidades del suelo y las correlaciones con los parámetros fisicoquímicos. Para ello se recolectaron muestras de suelo de 15 puntos aleatorios en un bosque reservado y se analizaron propiedades fisicoquímicas y microbiológicas a dos profundidades (10 y 20 cm). Se cuantificaron los siguientes microorganismos funcionales: aerobios viables, lactobacilos, actinomicetos, hongos y bacterias fijadoras de nitrógeno utilizando protocolos específicos. Se empleó la prueba no paramétrica de Wilcoxon para determinar diferencias entre profundidades y el coeficiente de correlación de Pearson (|ρ| ≥ 0,8) para analizar correlaciones entre parámetros fisicoquímicos y microorganismos. Los resultados sugieren que la abundancia de microorganismos aerobios viables, Lactobacillus, Actinomicetos y Fungi disminuye con la profundidad del suelo, mientras que las bacterias fijadoras de nitrógeno no muestran diferencias significativas. La disminución de la abundancia se relaciona con factores como la disponibilidad de nutrientes y oxígeno y las condiciones microambientales. Existe una correlación positiva fuerte entre la cantidad de microorganismos aerobios viables y Lactobacillus con el contenido de fósforo, y una correlación negativa entre Actinomicetos y el contenido de Arcilla. Los Actinomicetos prefieren suelos arenosos, mientras que los Fungi prosperan en suelos ricos en materia orgánica. La correlación negativa entre Fungi y Actinomicetos sugiere competencia por recursos limitados y posibles interacciones químicas, como la producción de compuestos antibióticos por parte de los Actinomicetos. Este estudio demuestra que la comprensión detallada de la microbiota del suelo puede ofrecer nuevas oportunidades para mejorar las prácticas agrícolas, especialmente en términos de gestión de la fertilidad del suelo y sostenibilidad ambiental.

Modulation of Broiler Intestinal Changes Induced by Clostridium perfringens and Deoxynivalenol through Probiotic, Paraprobiotic, and Postbiotic Supplementation.

Deoxynivalenol (DON) is a predisposing factor for necrotic enteritis. This study aimed to investigate the effects of a DON and Clostridium perfringens (CP) challenge on the intestinal morphology, morphometry, oxidative stress, and immune response of broilers. Additionally, we evaluated the potential of a Lactobacillus spp. mixture as an approach to mitigate the damage induced by the challenge. One-day-old broiler chickens (n = 252) were divided into seven treatment groups: Control, DON, CP, CP + DON, VL (DON + CP + viable Lactobacillus spp. mixture), HIL (DON + CP + heat-inactivated Lactobacillus spp. mixture), and LCS (DON + CP + Lactobacillus spp. mixture culture supernatant). Macroscopic evaluation of the intestines revealed that the CP + DON group exhibited the highest lesion score, while the VL and HIL groups showed the lowest scores. Microscopically, all Lactobacillus spp. treatments mitigated the morphological changes induced by the challenge. DON increased levels of reactive oxygen species (ROS) in the jejunum, and CP increased ROS levels in the jejunum and ileum. Notably, the Lactobacillus spp. treatments did not improve the antioxidant defense against CP-induced oxidative stress. In summary, a Lactobacillus spp. mixture, whether used as a probiotic, paraprobiotic, or postbiotic, exerted a partially protective effect in mitigating most of the intestinal damage induced by DON and CP challenges.

Rapid and Accurate Quantification of Viable Lactobacillus Cells in Infant Formula by Flow Cytometry Combined with Propidium Monoazide and Signal-Enhanced Fluorescence In Situ Hybridization.

Lactobacillus is an important member of the probiotic bacterial family for regulating human intestinal microflora and preserving its normalcy, and it has been widely used in infant formula. An appropriate and feasible method to quantify viable Lactobacilli cells is urgently required to evaluate the quality of probiotic-fortified infant formula. This study presents a rapid and accurate method to count viable Lactobacilli cells in infant formula using flow cytometry (FCM). First, Lactobacillus cells were specifically and rapidly stained by oligonucleotide probes based on a signal-enhanced fluorescence in situ hybridization (SEFISH) technique. A DNA-binding fluorescent probe, propidium monoazide (PMA), was then used to accurately recognize viable Lactobacillus cells. The entire process of this newly developed PMA-SEFISH-FCM method was accomplished within 2.5 h, which included pretreatment, dual staining, and FCM analysis; thus, this method showed considerably shorter time-to-results than other rapid methods. This method also demonstrated a good linear correlation (R2 = 0.9994) with the traditional plate-based method with a bacterial recovery rate of 91.24%. To the best of our knowledge, the present study is the first report of FCM combined with PMA and FISH for the specific detection of viable bacterial cells.

Tween 80™-induced changes in fatty acid profile of selected mesophilic lactobacilli.

Fatty acid profiles are crucial for the functionality and viability of lactobacilli used in food applications. Tween 80™, a common culture media additive, is known to influence bacterial growth and composition. This study investigated how Tween 80™ supplementation impacts the fatty acid profiles of six mesophilic lactobacilli strains (Lacticaseibacillus spp., Limosilactobacillus spp., Lactiplantibacillus plantarum). Analysis of eleven strains revealed 29 distinct fatty acids. Tween 80™ supplementation significantly altered their fatty acid composition. Notably, there was a shift towards saturated fatty acids and changes within the unsaturated fatty acid profile. While some unsaturated fatty acids decreased, there was a concurrent rise in cyclic derivatives like lactobacillic acid (derived from vaccenic acid) and dihydrosterculic acid (derived from oleic acid). This suggests that despite the presence of Tween 80™ as an oleic acid source, lactobacilli prioritize the synthesis of these cyclic derivatives from precursor unsaturated fatty acids. Myristic acid and dihydrosterculic acid levels varied across strains. Interestingly, palmitic acid content increased, potentially reflecting enhanced incorporation of oleic acid from Tween 80™ into membranes. Conversely, cis-vaccenic acid levels consistently decreased across all strains. The observed fatty acid profiles differed from previous studies, likely due to a combination of factors including strain-specific variations and growth condition differences (media type, temperature, harvesting point). However, this study highlights the consistent impact of Tween 80™ on the fatty acid composition of lactobacilli, regardless of these variations. In conclusion, Tween 80™ significantly alters fatty acid profiles, influencing saturation levels and specific fatty acid proportions. This work reveals key factors, including stimulated synthesis of lactobacillic acid, competition for oleic acid incorporation, and strain-specific responses to myristic and dihydrosterculic acids. The consistent reduction in cis-vaccenic acid and the presence of cyclic derivatives warrant further investigation to elucidate their roles in response to Tween 80™ supplementation.

Modulation of the gut microbiota and short-chain fatty acid production by gac fruit juice and its fermentation in in vitro colonic fermentation.

This study aimed to investigate the effects of gac fruit juice and its probiotic fermentation (FGJ) utilizing Lactobacillus paracasei on the modulation of the gut microbiota and the production of short-chain fatty acids (SCFAs). We conducted a comparison between FGJ, non-fermented gac juice (GJ), and control samples through in vitro digestion and colonic fermentation using the human gut microbiota derived from fecal inoculum. Our findings revealed that both GJ and FGJ led to an increase in the viability of Lactobacilli, with FGJ exhibiting even higher levels compared to the control. The results from the 16S rDNA amplicon sequencing technique showed that both GJ and FGJ exerted positive impact on the gut microbiota by promoting beneficial bacteria, notably Lactobacillus mucosae and Bacteroides vulgatus. Additionally, both GJ and FGJ significantly elevated the levels of SCFAs, particularly acetic, propionic, and n-butyric acids, as well as lactic acid, in comparison to the control. Notably, FGJ exhibited a more pronounced effect on the gut microbiota compared to GJ. This was evident in its ability to enhance species richness, reduce the Firmicutes to Bacteroidetes (F/B) ratio, promote Akkermansia, and inhibit pathogenic Escherichia coli. Moreover, FGJ displayed enhanced production of SCFAs, especially acetic and lactic acids, in contrast to GJ. Our findings suggest that the probiotic fermentation of gac fruit enhances its functional attributes in promoting a balanced gut microbiota. This beverage demonstrates potential as a functional food with potential advantages for sustaining intestinal health.

The alleviative effects of viable and inactive Lactobacillus paracasei CCFM1120 against alcoholic liver disease via modulation of gut microbiota and the Nrf2/HO-1 and TLR4/MyD88/NF-κB pathways.

Probiotics can alleviate alcoholic liver disease. However, whether inactive counterparts can produce similar outcomes requires further investigation. We investigated the effects of viable (V) and dead (D) Lactobacillus paracasei CCFM1120 on alcohol-induced ALD mice. The results showed that CCFM1120V and D ameliorated the disease symptoms and intestinal injury. Specifically, these interventions strengthened the intestinal barrier, as evidenced by the increased expression of ZO-1 (zonula occludens 1), occludin, and claudin-1 in the colon and the restored ileal microstructure, including the villi and crypts. In addition, they enhanced the antioxidant capacity of the liver by reducing the production of malondialdehyde and increasing the levels of glutathione and superoxide dismutase. The activation of Nrf2 and HO-1 may be responsible for recovering the antioxidant capacity. Interventions can decrease mouse TNF-α, IL-6 and IL-1β content in serum, probably through the TLR4/MyD88/NF-κB pathway. Furthermore, they possess the ability to restore the quantities of bacteria responsible for producing butyric acid, such as Lactobacillus, Blautia, Bifidobacterium, Ruminococcaceae, Faecalibaculum and Lachnospiraceae. Taken together, CCFM1120V and D apparently can modify the composition of the gut microbiota, foster the gastrointestinal equilibrium, fortify the intestinal barrier, augment the antioxidant capacity of the liver, and effectively shield it from ethanol-induced injury.

Cotton Fibers with a Lactic Acid-Like Surface for Re-establishment of Protective Lactobacillus Microbiota by Selectively Inhibiting Vaginal Pathogens.

Failure to reconstruct the Lactobacillus microbiota is the major reason for the recurrence of vaginal infection. However, most empiric therapies focus on the efficacy of pathogen elimination but do not sufficiently consider the viability of Lactobacillus. Herein, cotton fibers with a lactic acid-like surface (LC) are fabricated by NaIO4 oxidation and L-isoserine grafting. The lactic acid analog chain ends and imine structure of LC can penetrate cell walls to cause protein cleavage in Escherichia coli and Candida albicans and inhibit vaginal pathogens. Meanwhile, the viability of Lactobacillus acidophilus is unaffected by the LC treatment, thus revealing a selective way to suppress pathogens as well as provide a positive route to re-establish protective microbiota in the vaginal tract. Moreover, LC has excellent properties such as good biosafety, antiadhesion, water absorption, and weight retention. The strategy proposed here not only is practical, but also provides insights into the treatment of vaginal infections.

Chemical Composition and Antimicrobial Activity of Lavender (Lavandula angustifolia Mill.), Peppermint (Mentha piperita L.), Raspberry Seed (Rubus idaeus L.), and Ylang-Ylang (Cananga odorata (Lam.) Essential Oils—Towards Hurdle Technologies in the Production of Chocolate Mousse

The growing consumer demand for the development of functional foods with a number of benefits for the consumer has led to a considerable increase in the studies focused on examining different natural agents to be included in the composition of newly developed functional foods. The chemical compositions of the essential oils (EOs) of lavender (Lavandula angustifolia Mill.), peppermint (Mentha piperita L.), raspberry seed (Rubus idaeus L.), and ylang-ylang (Cananga odorata (Lam.)) were determined using gas chromatography with a mass selective detector (GC-MS). The antibacterial and antifungal activities of these EOs were examined using a high-throughput 96-well microplate bioassay procedure and the MICs of each EO against each test microorganism were determined. The results indicated significant antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538, Salmonella abony NTCC 6017, Pseudomonas aeruginosa NBIMCC 1390, Bacillus subtilis ATCC 19659, Penicillium chrysogenum ATCC 28089, Fusarium moniliforme ATCC 38932, Aspergillus niger ATCC 1015, and Aspergillus flavus ATCC 9643. To explore their potential applications in food preservation, model chocolate mousse food emulsions were prepared that incorporated EOs and/or selected probiotic lactobacilli strains in both free and encapsulated forms. The inclusion of EOs and/or probiotic lactobacilli resulted in enhanced microbial safety and an extended shelf life. Furthermore, the chocolate mousse variants that were biopreserved with the inclusion of probiotic lactobacilli maintained a high viable lactobacillus cell concentration throughout the storage period. As a result, these products would not only be suitable as functional probiotic foods but also as effective delivery vehicles for probiotic lactobacilli.

Sustainable starch-based edible films with agrifood residues as potential carriers for the probiotic Lactobacillus rhamnosus

Edible films are promising carriers for probiotics and can be composed by agrifood residues, which are usually rich in polymers and bioactive compounds. In this work, starch-based films were enriched with three types of agrifood residues (quince, potato and orange peels) and the incorporation of the probiotic Lactobacillus rhamnosus was studied, as well as the addition of inulin as a protective prebiotic. The resulting films were characterized in terms of mechanical properties, physicochemical properties, lactobacilli viability and microbiological properties. The mechanical properties of the films generally decreased with the introduction of L. rhamnosus, although this was highly dependent on the film composition. All films exhibited water vapor permeabilities in the typical range of starch-based films and were not greatly affected by the inclusion of probiotics. The loss of probiotic viability during films production was strongly related to the pH of the film-forming solutions. Films with agrifood residues had a slower loss of probiotic viability during storage, when compared to plain starch films, which may be explained by the presence of antioxidant compounds. Inulin was expected to improve viability, but this was not observed. Microbiological analysis showed that agrifood residues powders contained natural contaminant bacteria that were partially eliminated during film formation. Moreover, none of the target foodborne pathogens were detected in the analyzed samples. Overall, the results suggest that edible films containing agrifood residues can be a promising material for the delivery of probiotics and/or as primary packaging for some food products.

Nanocurcumin and viable Lactobacillus plantarum based sponge dressing for skin wound healing

Curcumin loaded solid lipid nanoparticles (CSLNs) and probiotic (Lactobacillus plantarum UBLP-40; L. plantarum) were currently co-incorporated into a wound dressing. The combination with manifold anti-inflammatory, anti-infective, analgesic, and antioxidant properties of both curcumin and L. plantarum will better manage complex healing process. Recent reports indicate that polyphenolics like curcumin improve probiotic effects. Curcumin was nanoencapsulated (CSLNs) to improve its bioprofile and achieve controlled release on the wound bed. Bacteriotherapy (probiotic) is established to promote wound healing via antimicrobial activity, inhibition of pathogenic toxins, immunomodulation, and anti-inflammatory actions. Combination of CSLNs with probiotic enhanced (560%) its antimicrobial effects against planktonic cells and biofilms of skin pathogen, Staphylococcus aureus 9144. The sterile dressing was devised with selected polymers, and optimized for polymer concentration, and dressing characteristics using a central composite design. It exhibited a swelling ratio of 412 ± 36%, in vitro degradation time of 3 h, optimal water vapor transmission rate of 1516.81 ± 155.25 g/m2/day, high tensile strength, low-blood clotting index, case II transport, and controlled release of curcumin. XRD indicated strong interaction between employed polymers. FESEM revealed a porous sponge like meshwork embedded with L. plantarum and CSLNs. It degraded and released L. plantarum, which germinated in the wound bed. The sponge was stable under refrigerated conditions for up to six months. No translocation of probiotic from wound to the internal organs confirmed safety. The dressing exhibited faster wound closure and lowered bioburden in the wound area in mice. This was coupled with a decrease in TNF-α, MMP-9, and LPO levels; and an increase in VEGF, TGF-β, and antioxidant enzymes such as catalase and GSH, establishing multiple healing pathways. Results were compared with CSLNs and probiotic-alone dressings. The dressing was as effective as the silver nanoparticle-based marketed hydrogel dressing; however, the cost and risk of developing resistance would be much lower currently.

High-speed cold centrifugation of milk modifies the microbiota, the ripening process and the sensory characteristics of raw-milk hard cheeses

The microbial population of raw milk plays a crucial role in the development of distinctive traits of raw-milk cheeses particularly appreciated by consumers. It was previously demonstrated that the microbial population of raw milk is modified by a high-speed centrifugation (also called bactofugation) conducted at 39 °C. The aim of the present study was to evaluate the effects of this process, performed once or twice, on the microbial, compositional, biochemical, and sensory characteristics of the derived hard cheeses. Experimental and control cheesemaking were conducted in parallel at a cheese factory during a 13-month period. Cheeses were analysed after 9, 15 and 20 months of ripening for microbial count, composition, proteolysis extent, volatile compounds, and sensory profile. Results evidenced that experimental cheeses were characterized by lower numbers of viable lactobacilli respect to control. Experimental cheeses also showed differences in the progress of primary and secondary proteolysis which, in turn, caused different patterns of free amino acids at all ripening times. Experimental cheeses had significantly lower content of esters and were differentiated from control for some traits by assessors. In conclusion, use of high-speed centrifugation of milk shall be discouraged if characteristic traits of raw-milk cheeses, particularly PDO cheeses, want to be retained.

Beneficial Effects of Viable and Heat-Inactivated Lactobacillus rhamnosus GG Administration on Oxidative Stress and Inflammation in Diet-Induced NAFLD in Rats

Oxidative stress and inflammation are well-known triggers of NAFLD onset and progression. The aim of this study is to compare the potential benefits of a viable probiotic (Lactobacillus rhamnosus GG) and its parabiotic (heat-inactivated) on oxidative stress, inflammation, DNA damage and cell death pathways in the liver of rats featuring diet-induced NAFLD. The consumption of the steatotic diet led to increased final body and liver weights, higher hepatic triacylglycerol content, altered serum transaminase levels and enhanced oxidative and inflammatory status. Administration of the probiotic and the parabiotic partially prevented the body weight increase induced by the steatotic diet, whereas the probiotic caused more effective decreasing hepatic triglyceride content. Sharp but nonstatistically significant decreases in serum transaminase levels were also observed for both treatments. The reduction in antioxidant enzyme activities found in the nontreated animals fed the steatotic diet was partially prevented by both treatments (GPx activity). Similarly, the reductions in nonenzymatic antioxidant protection (GSH content) and total antioxidant capacity (ORAC) found in the nontreated rats were restored by the administration of both treatments. These results show that both viable and heat-inactivated Lactobacillus rhamnosus GG administration partially prevent steatotic diet-induced liver oxidative stress and inflammation induced in rats.

SATELLITE IMAGE-DERIVED NEIGHBORHOOD CHARACTERICS EXPLAIN REGIONAL VARIATION IN CARDIOMETABOLIC DISEASE

We are aware of no study indicating the effects of synbiotic food consumption on metabolic profiles, inflammation and oxidative stress among diabetic patients. The aim of the current study was to investigate the effects of synbiotic food consumption on metabolic profiles, hs-CRP and biomarkers of oxidative stress in diabetic patients.This randomized double-blinded cross-over controlled clinical trial was performed among 62 diabetic patients aged 35–70 y. After a 2-wk run-in period, subjects were randomly assigned to consume either a synbiotic (n = 62) or control food (n = 62) for 6 weeks. A 3-week washout period was applied following which subjects were crossed over to the alternate treatment arm for an additional 6 weeks. The synbiotic food consisted of a probiotic viable and heat-resistant Lactobacillus sporogenes (1 × 107 CFU), 0.04 g inulin (HPX) as prebiotic with 0.38 g isomalt, 0.36 g sorbitol and 0.05 g stevia as sweetener per 1 g. Control food (the same substance without probiotic bacteria and prebiotic inulin) was packed in identical 9-gram packages. Patients were asked to consume the synbiotic and control foods three times a day. Fasting blood samples were taken at baseline and after a 6-wk intervention to measure metabolic profiles, hs-CRP and biomarkers of oxidative stress.Consumption of a synbiotic food, compared to the control, resulted in a significant decrease in serum insulin levels (changes from baseline: −1.75 ± 0.60 vs. +0.95 ± 1.09 μIU/mL, P = 0.03). Although we failed to find a significant effect of synbiotic food consumption on total- and LDL-cholesterol levels and HOMA-IR, the effects on FPG (22.3 vs. 4.2 mg/dL, P = 0.09), serum triglycerides (45.9 vs. 20.6 mg/dL, P = 0.08) and HDL-cholesterol levels (3.1 vs. −2 mg/dL, P = 0.06) tended to be significant. A significant reduction in serum hs-CRP levels (−1057.86 ± 283.74 vs. 95.40 ± 385.38 ng/mL, P = 0.01) was found following the consumption of synbiotic food compared with the control group. Supplementation with the synbiotic food led to a significant increase in plasma total GSH (319.98 vs. 19.73 μmol/L, P < 0.001) and serum uric acid levels (+0.7 vs. −0.1 mg/dL, P = 0.04) compared to the control food. No significant effect of the synbiotic food was observed on plasma TAC levels.In conclusion, consumption of a synbiotic food for 6 weeks among diabetic patients had significant effects on serum insulin, hs-CRP, uric acid and plasma total GSH levels.Clinical trial registration number: www.irct.ir: IRCT201201195623N1.