Casei Bacteria Research Articles

Exploring the Anti-Inflammatory and Antioxidative Potential of Selenium Nanoparticles Biosynthesized by Lactobacillus casei 393 on an Inflamed Caco-2 Cell Line.

Selenium (Se) plays a crucial role in modulating inflammation and oxidative stress within the human system. Biogenic selenium nanoparticles (SeNPs) synthesized by Lactobacillus casei (L. casei) exhibit anti-inflammatory and anti-oxidative properties, positioning them as a promising alternative to traditional supplements characterized by limited bioavailability. With this context in mind, this study investigates the impact of selenium and L. casei in ameliorating inflammation and oxidative stress using a cell line model. The study is centered on the biosynthesis of selenium nanoparticles (SeNPs) by L. casei 393 under anaerobic conditions using a solution of sodium selenite (Na2SeO3) in the bacterial culture medium. The generation of SeNPs ensued from the interaction of L. casei bacteria with selenium ions, a process characterized via transmission electron microscopy (TEM) to confirm the synthesis of SeNPs. To induce inflammation, the human colonic adenocarcinoma cell line, Caco-2 was subjected to interleukin-1 beta (IL-1β) at concentrations of 0.5 and 25 ng/ml. Subsequent analyses encompass the evaluation of SeNPs derived from L. casei, its supernatant, commercial selenium, and L. casei probiotic on Caco2 cell line. Finally, we assessed the inflammatory and oxidative stress markers. The assessment of inflammation involved the quantification of NF-κB and TGF-β gene expression levels, while oxidative stress was evaluated through the measurement of Nrf2, Keap1, NOX1, and SOD2 gene levels. L. casei successfully produced SeNPs, as confirmed by the color change in the culture medium and TEM analysis showing their uniform distribution within the bacteria. In the inflamed Caco-2 cell line, the NF-κB gene was upregulated, but treatment with L. casei-SeNPs and selenium increased TGF-β expression. Moreover, L. casei-SeNPs upregulated SOD2 and Nrf2 genes, while downregulating NOX1, Keap1, and NF-κB genes. These results demonstrated the potential of L. casei-SeNPs for reducing inflammation and managing oxidative stress in the Caco-2 cell line. The study underscores the ability of L. casei-SeNPs to reduce oxidative stress and inflammation in inflamed Caco-2 cell lines, emphasizing the effectiveness of L. casei as a source of selenium. These insights hold significant promise for the development of SeNPs derived from L. casei as potent anti-inflammatory and anti-cancer agents, paving the way for novel therapeutic applications in the field.

Pengaruh Suhu Penyimpanan Terhadap Kualitas Organoleptik Minuman Probiotik Susu Biji Nangka (Artocarpus heterophyllus)

The seeds of the jackfruit (Artocarpus heterophyllus) have not been used more by the community to make various processed food products. Even though jackfruit seeds contain many nutrients, especially potassium. Jackfruit seeds are rich in nutrients, especially carbohydrates, potassium, phosphorus and fat. The jackfruit seed contains 165 kcal of energy and 36.7 kcal of carbohydrates, which is quite high compared to the content of young jackfruit and ripe jackfruit. Seeing the content of jackfruit seeds which are rich in nutrients, these jackfruit seeds can be an alternative as a raw material for processing vegetable milk. Vegetable milk can be used as a basic ingredient for making probiotic drinks to increase the value of the benefits of this vegetable milk. This study used the Lactobacillus casei bacteria which acts as a probiotic bacteria. The study was conducted with different treatments at 20°C, 25°C, 30°C, and 37°C. Each treatment was repeated 4 times and 24 experimental samples were obtained. Based on the results of the study, it was shown that storage temperature had a significant effect on the organoleptic quality of probiotic jackfruit seed milk in terms of color and taste, but had no significant effect on the aroma parameters of probiotic jackfruit seed milk.

THE EFFECT OF PSYLLIUM FIBER AND CHOKEBERRY FIBRE ADDITION ON THE QUALITY OF PROBIOTIC FERMENTED MILK

Background. Fiber from fruit, vegetables and other sources of fiber can increase gut microbiota diversity and show anti-inflammatory activity. The use of fiber in the manufacture of probiotic fermented milk has excellent potential, as it can offer several benefits for both consumers and producers. In addition to its well-known physiological benefits, dietary fiber may reduce the risk of diseases of civilization. Consumers expect that current foods are health-promoting. Consumer awareness is increasing through an expanding variety of products and advertising campaigns favoring health-promoting foods, a trend that food manufacturers are also taking advantage of. The aim of the study was to determine the effect of psyllium fiber and chokeberry fiber addition in combination with probiotic cultures Bifidobacterium animalis ssp. lactis BB-12 and Lacticaseibacillus casei 431 on the quality of fermented milk. To investigate the effect of fiber on fermented milk quality, acidity, texture properties, syneresis and colorcolor were measured, moreover, an organoleptic evaluation and microbiological analysis were performed. Results and conclusion. Natural additives such as chokeberry fiber and psyllium fiber can be used in the production of milk fermented by Bifidobacterium BB-12 and L.casei. Psyllium fiber can be recommended for its syneresis-reducing properties. Whereas chokeberry fiber, due to its high polyphenol content, can be used as a natural colorcolorant and taste-enhancing additive. Adding fiber as chokeberry or psyllium positively stimulated the growth and survival of Bifidobacterium BB-12 and L. casei bacteria during storage. The hardness of the milk gel fermented by Bifidobacterium BB-12 or L. casei was significantly improved by adding psyllium fiber, increasing it by as much as about 2.5 N, and this trend was also maintained after 28 days of storage. Studies on using various fibers in fermented milk production should be continued to determine their health-promoting and technological potential.

Preparing a Synbiotic Jelly Product Using Wild Sage Seed Mucilage, Carrageenan, and Free and Encapsulated Lactobacillus Casei Bacteria

Background and Objectives Synbiotic foods have beneficial effects on the health. This study aims to evaluate the survival of Lactobacillus casei (L. casei) bacteria in the jelly containing sage seed mucilage and carrageenan. Subjects and Methods In this study, after extracting the mucilage from wild sage seed, its 1% concentration and 2% sodium alginate (Carrageenan), as an encapsulating material, were used to protect L. casei bacteria. In this regard, five groups of jelly samples were prepared: Carrageenan alone, carrageenan + wild sage seed mucilage, carrageenan+wild sage mucilage+free L. casei, carrageenan+free L. casei, and carrageenan+carrageenan+encapsulated L. casei. The pH and acidity of the produced jelly samples were evaluated during 28 days of storage at 4°C. Results During 28 days of storage, the log reduction of the logarithmic cycle in the carrageenan+wild sage seed mucilage+encapsulated L. casei group (1.9±0.01) was significantly lower than in the carrageenan + free L. casei group (4.08±0.1). The log number of remaining encapsulated L. casei group bacteria at the end of 28 days was higher than the standard limit (6.65±0.2). The presence of mucilage in the jelly led to an increase in the viability of free L. casei bacteria (3.09±0.21). In the samples containing free L. casei bacteria, acidity increased and pH decreased significantly during the storage period (P<0.05), while in samples containing encapsulated probiotics, changes in acidity and pH during storage were not significant (P>0.05). Conclusion Prebiotic bacteria can survive better in the encapsulated form. The addition of sage seed mucilage, can increases the survival of L. casei bacteria in the jelly during 28 days of storage.

Tofu Liquid Waste Treatment Process Using Anaerobic Baffled Reactor with Cow Stool and Lactobacillus casei Addition

Tofu liquid waste causes ecosystem damage due to its high organic matter content. Anaerobic baffled reactor (ABR) is a waste treatment in the absence of oxygen where there is a sedimentation process using activated sludge. This process converts organic material into methane gas, ammonia and hydrogen sulfide. This study aims to determine the effect of addition of cow feces and Lactobacillus casei bacteria on decreasing levels of COD, TSS, VSS, pH and gas pressure. The research process was carried out in two stages of processing tofu liquid waste, namely the batch and continuous stages. Batch and continuous processes use an ABR reactor which has four chambers. Parameters measured include, COD, TSS, VSS, pH and Gas Pressure. The results obtained by the best batch process were obtained without the addition of bacteria, the ratio of cow feces and activated sludge was 1:4, the COD removal was 81.63%, the TSS removal was 1,300 mg/L, the VSS removal was 1,200 mg/L and the pH value final 8.0 for 6 days. The best processing time in the continuous process is 6 hours, the COD removal is 78.12%, the TSS removal is 600 mg/L, the VSS removal is 500 mg/L and the final pH value is 6.5. It can be concluded that the more cow feces added, the faster and greater the removal of COD, TSS, VSS and pH values ​​tended to increase in the treatment of adding bacteria.

Media Modification Lactobacillus casei Strain Shirota with Skim Milk Enriched Green Coconut Water and Corn Starch

Bacteria need a medium that can play an important role in their growth. One example of a medium is the medium De Mann Rogosa and Sharpe (MRS) which is a specific medium for the growth of lactic acid bacteria. The use of MRS media on an industrial scale is ineffective due to its relatively high price. The purpose of this study was to find out the effect of skim milk media enriched with green coconut water and cornstarch on the growth of Lactobacillus casei bacteria. The type of research conducted is experimental research with post test only control group design where there are two groups, namely the control group and the experimental group. The experimental group consisted of 3 types of samples, namely skim milk and green coconut water, skim milk and cornstarch, as well as skim milk and green coconut water and cornstarch. The comparison of treatment levels for each sample was 1:2 and 1:3 and analyzed, including bacterial growth curves, growth speed and bacterial generation time. From each concentration, the results found that the addition of green coconut water and cornstarch had a significant effect on the growth of Lactobacillus casei (P˂0.01) to the speed of growth and generation time. It was concluded that the higher the concentration added it can accelerate the growth of Lactobacillus casei.

Growth of Lactobacillus casei FNCC0900 in Media Based Umbi Porang Plant (Amorphophallus muelleri BI.)

Porang tuber (Amorphophallus muellerii BI.) Is a type of tuber that has a high enough glucomannan content of 67%. Glucomannan is very difficult to digest by humans directly so it takes the role of probiotics. L. casei bacteria FNCC0900 as a probiotic agent capable of utilizing glucomannan as a carbon source for growth. The purpose of this study was to determine the growth pattern and changes in environmental factors, namely the pH value of the probiotic bacteria L. casei FNCC0900 growth medium. The parameters in this study consisted of the highest cell density, generation time and pH value changes in Glucose Yeast Peptone Liquid Media, Porang Boiled Water Media and Porang Flour Liquid Media using the drop plate method which had 4 repeated calculations. Porang Boiled Water Liquid Media has a faster log phase period with a higher cell density than Porang Flour Liquid Media, but the shortest generation time is found in Porang Flour Liquid Media with the highest number of generations. L. casei FNCC0900 bacteria are more able to reduce the pH of Glucose Yeast Peptone Liquid Media compared to porang tuber-based media, so in this case L. casei FNCC0900 can be stated to be able to grow on porang tuber-based media with growth patterns, generation time, cell density and pH value. which varies.

The Effect of Extrusion Voltage and Flowrate to the Viability and Survivability of Probiotic <em>L. casei</em> Encapsulated in Alginate-Chitosan

Chitosan-coated L. casei containing alginate capsules (shortened as L. casei capsules) were prepared by extruding L. casei containing alginate solution at different extrusion voltage and and flow rate followed by coating the wet capsules in chitosan solution. This study aimed to determine the effect of extrusion voltage and sodium alginate liquid flow rate on the viability of L. casei bacteria in the encapsulation process. The encapsulation process in this study was carried out by the extrusion method using sodium alginate of 1% (w/v) and chitosan of 0.2% (w/v). The resulted beads were immersed in a simulated gastric fluid (SGF) (NaCl 0.2%; HCl 0.5 M with a pH of 1.5) for 1, 60, and 120 min at 37 °C. The number of L. casei cells before encapsulation was 12.3 log CFU. After encapsulation, the maximum viability of L. Casei obtained by voltage variations of 0 kV and flow rate 5 mL/min were 12.26 log CFU. After testing the beads in SGF for 1 min, the results obtained indicate that viability of L.casei in the sodium alginate - chitosan beads with an extrusion voltage of 0 kV and 5 mL/min was 11.8 log CFU/g. The result indicated that encapsulated L. casei in the sodium alginate - chitosan beads with a voltage of 0 kV and 5 mL/min was the highest survivability level of 97.38 %. The conclusions of the study were The higher extrusion voltage can kill more L. casei while the higher extrusion flow rate can protect more L. casei.

Effect of the Cellular Extract of Co-cultured Lactobacillus Casei on BAX and Human β-Defensin 2 Genes Expression in HT29 Cells

Aims: Defensins are cysteine-rich antimicrobial cationic peptides and BAX is a proapoptotic gene that can cause cell death. This study aimed to investigate the effect of cellular extract of co-cultured Lactobacillus casei on the expression of BAX and human β-defensin 2 (hBD-2) genes in HT29 cells. Methods & Materials: This experimental study was conducted in the Research Center for Pharmaceutical Nanotechnology of Tabriz University of Medical Sciences in 2017. The HT29 cell line was obtained from the Pasteur Institute of Iran, and cells were assessed using Microculture Tetrazolium Test (MTT) after culturing. DNA was extracted from the treated cells, and then the DNA ladder assay was carried out. After preparing cDNA, the expression levels of BAX and hBD-2 genes in the HT29 cell line were measured using a real-time Polymerase Chain Reaction (PCR) method. Findings: The results of the MTT assay indicated that Lactobacillus casei inhibited the proliferation of HT29 cells and induced apoptosis in these cells. Results of DAPI staining and DNA ladder assay obtained from treating HT29 cells by Lactobacillus casei showed qualitative changes in cell apoptosis. Moreover, realtime PCR results indicated that Lactobacillus casei bacteria significantly increased the expression of the hBD-2 gene in HT29 colon cancer cells within 12-24 hours (P= 0.023), while BAX gene expression showed no significant change in the first 24 hours (P= 0.37). Conclusion: The extract of Lactobacillus casei can be used to stimulate cancer cells to produce β-defensins, inhibit pathogens, prevent the stimulation of cellular signaling, and fight antibiotic-resistant bacteria.

Molecular Routes to Specific Identification of the Lactobacillus Casei Group at the Species, Subspecies and Strain Level.

The genus Lactobacillus includes, among others, Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus, species that are collectively referred to as the Lactobacillus casei group. Many studies have shown that strains belonging to this group may decrease lactose intolerance, the effects of inflammatory bowel disease, diarrhea, constipation, food allergies and even colon cancer. Moreover, evidences exists of positive effects of these bacteria on mucosal immunity and blood cholesterol level. Because of their beneficial influence on human health, many of them are used as food additives and probiotic pharmaceuticals. It should be stressed that health-promoting properties are not attributed at the species level, but to specific strains. Therefore, procedures are necessary to allow specific identification at each phylogenetic level—genus, species and strain. In this paper we present a practical overview of molecular methods for the identification and differentiation of L. casei bacteria. The research included 30 bacterial strains belonging to three species: L.casei, L. paracasei and L. rhamnosus. Among the tested procedures were genus- and species-specific PCR, multiplex-PCR, Real-Time HRM analysis, RFLP-PCR, rep-PCR, RAPD-PCR, AFLP-PCR, and proteomic methods such as MALDI-TOF MS typing and SDS-PAGE fingerprinting. The obtained results showed that multiplex-PCR and MALDI-TOF MS turned out to be the most useful methods to identify the tested bacteria at the species level. At the strain level, the AFLP-PCR method showed the highest discriminatory power. We hope that the presented results will allow for the easy selection of an appropriate procedure, depending on the experiment conducted and the equipment capabilities of any given laboratory.

The encapsulation of Lactobacillus casei probiotic bacteria based on sodium alginate and chitosan

Chitosan is a polysaccharide that can be used as a material in the encapsulation of probiotic bacteria. The process of bacterial encapsulation with chitosan polymers was carried out to protect the Lactobacillus casei bacteria which cannot last long in very acidic environments so that they can survive when exposed to gastric acid conditions and can live in the intestine. The purpose of this study was to determine the effect of extrusion voltage on the survivability of L. casei bacteria in the encapsulation process. The encapsulation process in this study was carried out by the extrusion-emulsion method using a sodium alginate of 1 % (w/v) and chitosan of 0.2 % (w/v) and voltage variations of 0 kV, 10 kV and 20 kV. The resulting beads were immersed in a simulated gastric fluid (SGF) (NaCl 0.2 %; HCl 0.5 M with a pH of 1.5) for 0, 60 and 120 minutes at 37 °C. The number of L. casei cells before encapsulation was 1 × 109 cfu/mL. After encapsulation with voltage variations of 0 kV, 10 kV and 20 kV, the viability of probiotics were 2.6 × 108 cfu/g, 1 × 107 cfu/g, and 1 × 103 cfu/g, respectively. After testing the beads in SGF, the obtained results indicated that viability of L.casei in the sodium alginate - chitosan beads with an extrusion voltage of 0 kV were 20,300 cfu/g, 10 kV were 30 cfu/g and 20 kV were 0 cfu/g. The results of these studies indicated that survivability of L. casei in the sodium alginate - chitosan beads with a voltage of 0 kV indicates the highest survivability level of 51.19 % of the number of cells encapsulated after incubation in SGF and the higher voltage can kill more L. casei.

Delipidation and Deproteination of Coconut Dregs Fermented with Aspergillus niger to Produce Prebiotic compounds

Coconut dregs are the waste product of oil extraction, either to produce conventional cooking oil or virgin coconut oil (VCO). These coconut dregs contain mannan compound in the form of galactomannan. Fermentation of this compound could generate mannan-oligosaccharides, having prebiotic properties, due to the presence of mannanase enzyme during fermentation. A study on fermentation of coconut dregs was carried out with a delipidation process using n-hexane and deproteination using NaOH 1M. The fermentation process was done using Aspergi l lus niger with different duration of fermentation (48, 72, 96, 120 and 144 hours). The fermentation product was placed in the free-air container to kill the fungi and keep the biodegradation process through enzymatic process for 0, 24, 48 and 72 hours. The products were tested for prebiotic properties through spread plate method using Lactobacillus casei and Escherichia coli . The results of the delipidation process could decreased lipid content to 0.77% and the deproteination process could bring down crude protein of coconut dregs to 1.45%. The highest mannanase activity was found when coconut dregs were fermented for 120 hours with a value of 1.0331U/ml. Hydrolyzate produced during incubation had a prebiotic activity by increasing the population Lactobacillus casei and decreasing population of Escherich i a coli . Lactobacillus casei bacteria increased from 2x10 7 CFU/ml to between 30.6x10 7 CFU/ml and 35.0x10 7 CFU/ ml, whereas Escherich i a coli bacteria decreased from 2x10 7 CFU / ml to between 5.3x10 6 CFU/ml and 8.3x10 6 CFU/ml. Keyword s : Coconut dregs, Aspergil l us niger , Prebiotics DOI: 10.7176/JBAH/10-6-03 Publication date: March 31 st 2020

Exploring antibacterial activity and hydrolytic stability of resin dental composite restorative materials containing chitosan.

The development of resin dental composite restorative materials (RDCRM) with antimicrobial activity may prevent the problem of secondary caries and consequently prolong longevity of RDCRM. This study explored antibacterial activity and hydrolytic stability of RDCRM containing chitosan. The antibacterial activity of microhybrid and flowable RDCRM containing 0, 0.25, 0.5 and 1% wt/wt chitosan (CS) against Lactobacilli casei bacteria was investigated using agar diffusion test and direct contact methods. Hydrolytic stability of experimental RDCRM was evaluated using gravimetric analysis. The control and experimental flowable and microhybrid RDCRM exhibited no growth inhibition zone in the lawn growth of Lactobacilli casei. The direct contact test revealed that colony forming unit count of Lactobacilli casei was comparable among the experimental and control RDCRM. The water sorption and solubility values of control as well as experimental flowable and microhybrid RDCRM were found to be within the ISO accepted range. There was statistically no significant difference in water sorption and solubility of different experimental RDCRM groups. The one percentage of CS into experimental RDCRM have no antibacterial activity against Lactobacilli casei. The hydrolytic stability of RDCRM containing CS was acceptable.

Fouling of nanofiltration membranes used for separation of fermented glycerol solutions

AbstractIn this study, the glycerol solutions were fermented using Lactobacillus casei bacteria. The broths were pre-treated by microfiltration, followed by a further separation with nanofiltration. The latter process was carried out in two stages, using the NF270 and NF90 membranes, respectively. The concentrates thus obtained were enriched with citric acid (first stage) and then with lactic acid and glycerol (second stage). By means of SEM and AFM microscopy, as well as ATR-FTIR analysis, the intensity of membrane-fouling was studied. The colloidal fouling and bio-fouling caused a more than two-fold decrease in the permeate flux during microfiltration of the broth. This pre-treatment stage was effective, and a permeate turbidity of less than 0.2 NTU was obtained. However, the nanofiltration membranes exhibited a 30 % flux decline over the course of the process, mainly due to the organic fouling.

Continuous cell from cell separation by traveling wave dielectrophoresis

AbstractIn this contribution we present a label-free continuous separation device for suspended-grown biological cells based on traveling wave dielectrophoresis (twDEP). Suspended-grown cells could otherwise only be separated by flow-cytometry which requires elaborate labeling steps. The chip consists of a structured PDMS layer on glass in which the channels have been defined. Parallel electrodes positioned along the microfluidic channel were used to expose selected cells to a twDEP force perpendicular to a pressure driven flow. With this device we have successfully separated viable Saccharomyces cerevisiae and Jurkat (T-cell leukemia) cells from a mix also containing cell culture debris, non-viable cells, and Lactobacillus casei bacteria.

Continuous separation of viable cells by travelling wave dielectrophoresis

In this contribution we present a label-free continuous separation device for suspended-grown biological cells based on travelling wave dielectrophoresis (twDEP). Suspended-grown cells could otherwise only be separated by flow-cytometry which requires elaborate labelling steps. The biochip consists of a structured PDMS layer on glass in which the channels have been defined. Parallel electrodes positioned along the microfluidic channel were used to expose selected cells to a twDEP force perpendicular to a pressure driven flow. With this device we have successfully separated suspended-grown viable Jurkat (T-cell leukaemia) cells from a mix also containing non-viable cells and L. casei bacteria.

Cross-effects of extracellular factors of adaptation to stress in Luteococcus casei and Saccharomyces cerevisiae

Saccharomyces cerevisiae yeasts (lower eukaryotes) were shown to produce a protein exometabolite with reactivation activity. We demonstrated cross-effects of extracellular protein factors of adaptation to stress (heat and UV irradiation) in yeasts and Luteococcus casei bacteria. The possibility for isolation and partial purification of protein exometabolites from the culture liquid of yeasts and bacteria by similar methods, as well as the similarity of elution profiles for the active proteins in high-performance liquid chromatography, suggests that the proteins (or fragments thereot) of the organisms studied are homologous.

Enhanced antigen-specific delayed-type hypersensitivity and immunoglobulin G2b responses after oral administration of viable Lactobacillus casei YIT9029 in Wistar and Brown Norway rats.

In this study, the effects of orally administered viable Lactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined. For this purpose, we used the Trichinella spiralis host resistance model. Two weeks before and during T. spiralis infection, rats were fed 10(9) viable L. casei bacteria 5 days per week. The T. spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L. casei. In both rat strains fed L. casei, serum T. spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased. In the model, no significant effects of L. casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed. Serum specific antibody responses, other than IgG2b, were not changed by feeding of L. casei. In contrast to L. casei, it was shown that orally administered Bifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model. Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L. casei. Furthermore, our data do not indicate that the effect of oral supplementation with L. casei is dependent on the genetic background of the host.