Culture Lactobacillus Plantarum Research Articles

Formulation and quality control of semi-solid containing harmless bacteria by-products: chronic wounds pro-healing activity

Chronic wounds are those that remain in a chronic inflammatory state and fail to follow normal healing process. Infection is one of the most important causes of chronicity. A frequent pathogen isolated from chronic infections is Pseudomonas aeruginosa; refractory to therapy and host immune attack in its biofilm phenotype. Lactobacillus plantarum cultures supernatants (LAPS) interfere with its pathogenic capacity. In addition, LAPS showed bacteriostatic and bactericide properties and is neither cytotoxic nor an inductor of necrosis-apoptosis. LAPSs chemical composition was determined; allowing us to propose a correlation between its constituents and their biological activity. This article shows a pharmaceutical dosage form designed by using LAPS as an API with pro-healing activity and its quality control. Pharmacotechnical and anti-microbial assays were adapted to demonstrate that the vehicle used does not modify LAPS activities. Selected formulation (F100) showed fair mechanical and technological properties. From the in vitro release assays was found an adequate release from the carrier matrix and maintains its anti-pathogenic activity for 6 months. We propose F100 for chronic wounds treatment. The use of harmless bacteria by-products, such as LAPS, to antagonize infectious pathogens that have ability to form biofilm is an efficient and economic approach to treat infected chronic wounds.

Microbiological and Nutritional Assessment of Starter-Developed Fermented Tigernut Milk

Fermented tigernut milk (FTM) was prepared from three different varieties (fresh yellow, big and small dry brown) of tigernut (Cyperus esculenta) obtained from Bodija market, Ibadan, Oyo state. Fifty two microbial isolates were obtained from FTM at different fermentation times, 32 of which were on MRS agar, 12 on nutrient agar and 8 on malt extract agar. Lactic acid bacteria were identified as Lactobacillus plantarum (LP), Lactococcus lactis (LC), Lactobacillus brevis, Lactococcus cremoris, Lactobacillus bulgaricus and Lactococcus thermophilus (LT). The non-LAB identified includes E. coli, Bacillus species and Proteus species while the yeasts include Saccharomyces cerevisiae and Candida kefyr. Lactic acid bacteria were found to predominate the total microflora of the FTM with their count ranging between 2.0 × 104 cfu/ml to 2.0 × 108 cfu/ml. Microbiological examination revealed that the FTM was safe for consumption as non-LAB counts were below the limit of acceptance which is 2.0 × 105 cfu/ml for dairy milk by Codex Alimentarius Commission. The quantity of lactic acid produced by the LAB isolates ranged between 0.86 g/l - 2.86 g/l while that of hydrogen peroxide ranged between 0.16 g/l - 0.51 g/l. Starter cultures were selected based on predominance of isolate, physiological characteristics, quantity of lactic acid and hydrogen peroxide production. The tiger nut varieties were fermented with the following starter combinations LP, LP/LC, LP/LC/LT, LP/LT while the fifth was spontaneously fermented. The nutritional, chemical and sensory properties of the starter fermented tiger nut milk were evaluated. The highest protein content (24.80%) was obtained in FTM with mixed cultures of LP/LC/LT while the least (3.00%) was obtained in spontaneously fermented milk. There was a significant difference in the FTM varieties. The highest fat content (9.40%) was obtained in spontaneously fermented tiger nut milk while the least (3.40%) was found in FTM with mixed cultures of LP/LC/LT. Fermentation decreased the pH and increased the lactic acid of the starter developed FTM, while sensory evaluation test showed that the FTM with mixed cultures of LP/LC/LT was highly acceptable. In conclusion, an excellent and acceptable FTM can be produced using treatments which comprise of pasteurization at 90° for 15 minutes, fermentation at 45℃ for 18 hours using mixed cultures of Lactobacillus plantarum, Lactococcus lactis and Lactococcus thermophilus.

Lactobacillus plantarum 29 Inhibits Penicillium spp. Involved in the Spoilage of Black Truffles (Tuber aestivum)

The effect of an antifungal culture of Lactobacillus plantarum to be used in the storage at refrigeration temperature of fresh black truffles was examined. The strain was selected among 29 lactobacilli isolated from foods and evaluated for their viability and acidification activity at 4 °C, as well as for their inhibitory activity against 11 Penicillium strains isolated from truffles stored at refrigeration temperature. Lb. plantarum 29 showed the ability to hold not only the growth of Penicillium isolated from truffles, but also that of P. digitatum DSM 2750, a green mold involved in the spoilage of truffles. The antifungal activity was observed in vitro and in situ, and the sensory characteristics of truffles were preserved during the cold storage.

Identification of Antioxidants Produced byLactobacillus plantarum

We identified two compounds that demonstrated 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity from cultures of Lactobacillus plantarum. Spectroscopic analyses proved these compounds to be L-3-(4-hydroxyphenyl) lactic acid (HPLA) and L-indole-3-lactic acid (ILA). The respective EC50 values for HPLA and ILA were 36.6 ± 4.3 mM and 13.4 ± 1.0 mM.

The Effect of a Mixed-Starter Culture of Lactic Acid Bacteria on the Characteristics of Pickled Orange-Fleshed Sweet Potato L.) (Ipomoea batatas

In this study, fermentation process was carried out on orange-fleshed sweet potato cubes to produce sweet potato pickle using a mixed culture of Lactobacillus plantarum and Leuconostoc mesenteroides at 30 °C over 12 days period. Spontaneous fermentation was also performed as a control. Samples were withdrawn at various time intervals for analyses of reducing sugar content, total number of lactic acid and non-lactic acid bacteria, lactic acid concentration, pH, and sensory attributes. The results showed that using a mixed culture of L. plantarum and L. mesenteroides could greatly reduce contamination of non-lactic acid bacteria, retaining low amount of reducing sugar, rapidly producing lactic acid and consequently decreasing pH value of the pickle, as well as giving better sensory score. After 12 d of fermentation, sample of pickle inoculated with mixed culture showed the following characters: total lactic acid content 0.5%, total lactic acid bacteria 8.46 log10 CFU mL-1, total non-lactic acid bacteria 1 log10 CFU mL-1, total reducing sugar 0.84 g L-1, texture 64.92 mm 50 g-1 s-1, and hedonic sensory score for both taste and aroma 4 (like) in a scale of 5. These results indicated the potential ability of the mixed culture of lactic acid bacteria to improve the quality of the pickle fermented spontaneously.

Effect of inoculation of selected Staphylococcus xylosus and Lactobacillus plantarum strains on biochemical, microbiological and textural characteristics of a Tunisian dry fermented sausage

In this study, a selected starter culture of Lactobacillus plantarum and Staphylococcus xylosus was used for the manufacture of a traditional Tunisian dry fermented sausage. Changes of microbiological, biochemical, sensorial and textural characteristics during the ripening of sausages were investigated.Counts of lactic acid bacteria and staphylococci were significantly higher in inoculated sausages than in control ones. Moreover, the acidifying activity of the selective L. plantarum strains improved the hygienic quality of fermented sausages by reducing counts of Enterobacteriaceae. Both a significant decrease in pH and a significant increase in nitrite contents were observed in inoculated sausages compared to control ones. The loss of moisture during the ripening of sausages was not affected by the use of starters. Proteolysis and lipolysis were observed both in control and in inoculated sausages; however, these activities during ripening could be due to the activity of starter culture enzymes or the activity of meat endogenous enzymes. The Factorial Discriminative Analyze (FDA) of sensorial parameters (aroma, color, taste and firmness) discriminate clearly inoculated sausages from non-inoculated ones. In fact, inoculated sausages showed a more intensive red color, related to the production of nitrosomyoglobin pigment, and an acid flavor. Finally, textural parameters of sausages were not affected by the use of the selective starters.

Effect of different starter cultures on the biogenic amines production as a critical control point in fresh fermented sausages

The selection of critical control points is one of the most important steps in the design of a hazard analysis and critical control points system. In this study, the influence of starter culture inoculation on biogenic amine accumulation in fresh fermented sausage as critical control point was studied. The effect of three starter culture Lactobacillus plantarum plus Bifidobacterium lactis, L. plantarum plus Bifidobacterium bifidum or L. plantarum plus B. lactis and B. bifidum on amines production were investigated. Tyramine (TY), Cadaverine (CA), Putrescine (PU), Histamine (HIS) and Tryptamine (TR) contents increased during the fermentation and storage stage, and TY was the prevailing amine in the final product. Sausages produced by fermentation with starters, as compared to natural fermentation (control), had a lower amount of biogenic amine. A great reduction in biogenic amine content was achieved when Bifidobacterium sp. were inoculated, also levels of species of Enterobacteriaceae and Pseudomonas decreased, while lactic acid bacteria counts increased during fermentation and storage.

Effect of water activity and protective solutes on growth and subsequent survival to air-drying of Lactobacillus and Bifidobacterium cultures

Probiotic cultures of Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus casei and Lactobacillus acidophilus were grown in media having water activities (a (w)) adjusted between 0.99 and 0.94 with NaCl or with a mixture of glycerol and sucrose in order to find conditions of osmotic stress which would still allow for good growth. Cultures grown at a (w) = 0.96 or 0.99 were then recovered by centrifugation, added to a sucrose-phosphate medium and air-dried. In some assays, a 2-h osmotic stress was applied to the cell concentrate prior to air-drying. Assays were also carried out where betaine, glutamate and proline (BGP) supplements were added as protective compounds to the growth or drying media. For most strains, evidence of osmotic stress and benefits of BGP supplementation on growth occurred at a (w) = 0.96. Growing the cells in complex media adjusted at a (w) = 0.96 did not enhance their subsequent survival to air-drying, but applying the 2-h osmotic stress did. Addition of the BGP supplements to the growth medium or in the 2-h stress medium did not enhance survival to air-drying. Furthermore, addition of BGP to a sucrose-phosphate drying medium reduced survival of the cultures to air-drying. This study provides preliminary data for producers of probiotics who wish to use air-drying in replacement of freeze-drying for the stabilization of cultures.

Cassava solid-state fermentation with a starter culture of Lactobacillus plantarum and Rhyzopus oryzae for cellulase production

A mixture of two microorganisms was used as starter culture, with the aims of producing cellulose from fermented cassava roots. Cellulase yields of 430 IU/g were recorded in solid-state fermentation of cassava starch with mixed culture of Lactobacillus plantarumand Rhyzopus oryzae, isolated from cassava dried chips. High cellulase activity (8.6 IU/ml) per unit volume of enzyme broth and high yields of cellulases were obtained and attributed to the growth of both strains on cellulose fraction of cassava roots. In Solid State Fermentation with Starter mix-culture (SSFS), fermentation retting was completed in two days, and significant softening was noticed. No significant softening was evidence in Solid State Sterile Fermentation (SSSF) and Liquid state fermentation (LSF). Keywords: Cassava, cellulases, Lactobacillus plantarum, Rhyzopus oryzae, solid state fermentation, liquid state fermentation.

Enhancement of γ-aminobutyric acid in a fermented red seaweed beverage by starter culture Lactobacillus plantarum DW12

Lactobacillus plantarum DW12, a gamma-aminobutyric acid (GABA) producing strain, was used as a starter culture to produce a functional fermented red seaweed beverage (FSB). Optimal conditions for producing FSB were established using Central Composite Design by varying the amounts of monosodium glutamate (MSG), sucrose and the initial pH in MRS medium. After a verification test, 1% MSG, 6% sucrose and an initial pH of 6 were selected. Four treatments were tested: traditional formula (A), red seaweed-cane sugar-potable water = 3:1:10, w/w/v, initial pH 6; the traditional formula with a 5% starter culture consisting of 4.1 x 10 9 CFU of DW 12/ml (B); formula A modified by changing the amounts of cane sugar and MSG to 6% and 1%, respectively (C); formula C with a 5% starter culture added (D). Comparison among the 4 treatments showed that the treatment D produced the highest amount of GABA (4000 mg/L) during days 45-60 while the GABA content of A, B and C treatments was 340, 730 and 1690 mg/L, respectively. However, the results of the sensory test for the treatments C and D showed that the presence of MSG produced an unsatisfactory salty taste. All finished products from the 4 treatments met Thai standard guidelines for chemical and microbiological qualities after 120 days. The results indicated that enrichment of the GABA content in FSB is possible by adding MSG and the GABA producing strain DW12; however, the appropriate amount of MSG addition should be further studied.

Protective effect of Lactobacillus plantarum against Salmonella typhimurium in mice.

Background: Probiotics are defined as living microbial food supplements that beneficially affect the host by improving its intestinal balance. In recent years there is a great interest for viable microorganisms that promote or support a beneficial balance of microbial population of gastrointestinal tract. Probiotics may be consumed in different forms, comprising foods, mainly in fermented state and pharmaceutical products, mainly as capsules or in microencapsulated forms.Methods: Twelve albino mice were divided into four groups designated as 1, 2, 3 and 4. Each group consists of 3 mice, and subject to the followings treatments: Group 1: This group was used as a control. Group 2: This group was dosed with 0.1ml. of 0.5 × 105 cfu/ml of Lactobacillus plantarum culture. Group 3: This group was dosed with 0.1ml. of 0.5 × 105 cfu/ml Salmonella typhimurium culture. Group 4: this group was dosed with 0.1ml of 109cfu/ml Lactobacillus plantarum culture, and infected with 0.1ml of 0.5 × 105cfu/ml culture of Salmonella typhimurium, then histological sections were made from intestine and examined to evaluate protective effect of Lactobacillus plantarum.Results: Histopathological study exhibited that intestinal sections of mice infected with Salmonella typhimurium showed a necrosis, degenerative changes and inflammatory cells infiltration as compared with normal sections taken from uninfected mice, while treating with Lactobacillus plantarum prevented the histopathological effect of Salmonella typhimurium on mice intestine.Conclusion: Probiotic L. plantarum had a protective effect on intestine sections of mice infected with Salmonella typhimurium

Glycemic index and phenolics of partially-baked frozen bread with sourdough

Different lactic acid bacteria starters were used to prepare sourdough to make partially-baked frozen wholemeal wheat bread. The sourdough was prepared with a pure culture of Lactobacillus plantarum or with commercial starters containing Lactobacillus brevis combined with Saccharomyces cerevisiae var. chevalieri (LV4), Lactobacillus fermentum (PL1), or Lactobacillus fermentum with phytase (PL3). We determined the acetic and lactic acid concentrations in sourdough, bread chemical composition, total phenolics content and glycemic index (GI) in vivo. Depending on the starter, the lactic to acetic acid ratio in the sourdough was significantly different. The GI of control bread without sourdough (70) was significantly higher than that of bread containing sourdough prepared with LV4 starter (50), PL1 starter (56) or PL3 starter (56), but not from bread with L. plantarum sourdough (60). The addition of 10% sourdough with a lower molar ratio of lactic to acetic acid ( ≤ 4) and higher total phenolics content is preferable for generating bread with medium and low GI.

Pengaruh Penggunaan Starter Bakteri Asam Laktat Lactobacillus plantarum dan Lactobacillus fermentum terhadapTotal Bakteri Asam Laktat, Kadar Asam dan Nilai pH Dadih Susu Sapi

The objective of the current study to determine the effect of the use of bacteria Lactobacillus plantarum and Lactobacillus fermentum in the manucfacture of cow’s milk curd on the total lactid acid bacteria, acidity and pH value of cow’s milk curd. The experiment use fresh cow milk and starter culture of Lactobacillus plantarum, and L. plantarum. Starter culture added about 3% from milk volume than incubated at room temperature during 48 hours. The treatments were fermentation with L. plantarum (A1); L. fermentum (A2); L. plantarum + L. fermentum (A3). There were significant effect of addition of starter L. plantarum (A1), L. fermentum (A2), and the combination of L. plantarum + L. fermentum (A3) on total lactic acid bacteria and acidity, however, there was no statistically significant on the pH value of cow's milk curd. It was concluded that using of starter culture combinations of Lactobacillus plantarum + Lactobacillus fermentum (A3) increases the total lactic acid bacteria and acid levels of cow’s milk curd.

News in wound healing and management

Nonhealing wounds are a significant problem in the healthcare system all over the world. The present review focuses on some recent developments and promising clinical progresses in wound management. New findings have increased our knowledge in several wound areas. In the treatment of wounds, the new trend in the wound device marked is to produce dressings containing compounds or drugs. This could be local antiseptics (silver, other antiseptics, honey) and pain relieving drugs such as ibuprofen and morphine. New treatments such as anti-tumor necrosis factor alfa (anti-TNFalpha) and Lactobacillus plantarum cultures have also been successfully used in hard to heal, atypical wounds. Knowledge on influencing factors as smoking and biofilm on the healing process has also been improved. Smoking results in delayed healing and increased risk of postoperative infection, whereas the role of biofilm is still at an exploratory level. Organizing models for optimal wound management are constantly being developed and refined. Recent knowledge on the importance of new dressing materials containing active substances, new treatments for atypical wounds, influencing factors on the healing process and organization in the wound area are increasingly been launched. This may in the coming years significantly improve the treatment outcome of problem wounds.

Differences between Pseudomonas aeruginosa in a clinical sample and in a colony isolated from it: Comparison of virulence capacity and susceptibility of biofilm to inhibitors

We study the differences between Pseudomonas aeruginosa from an infected wound (clinical strain) and a colony isolated from it. We assessed the in vitro inhibition of these P. aeruginosa biofilms by DNase and filtrate of Lactobacillus plantarum cultures (acid=AF and neutralize=NF) with crystal violet technique. Inhibition by AF was greatest than DNase for clinical and isolated strain (p<0.001) and greatest than NF for clinical (p<0.05) and isolated strain (p<0.001). Using a burn model in mice, we compared the infection producing by clinical and isolated strains in planktonic and biofilm form. Deaths were quantified and the infection was assessed by determining CFU/g of tissue in the lesion, spleen and liver. The infections with planktonic bacteria tended to become systemic and more deadly than biofilm infections. All infected wounds required the same healing period (30 days). These findings were independent of the origin of the bacteria (clinical or colony isolated strain).

Survey of the improvement of fish fermentation for lanhouin production in Benin

The objective of this study was to control ‘maturation’ process of fish during the fermentation, by using bio-organisms that can give the expected improvement. Starter culture of Lactobacillus plantarum was used as bio-organism on species of fish such as Pseudotolithus senegalensis and Scomberomorus tritor. The inoculum was serially diluted (10–1 to 10–5) in sterile Ringer solution. The result of this improved method showed significantly a product with a mild odour compared to odour from lanhouintraditionally obtained by chance fermentation. Key words: Fermentation, fish, Lactobacillus plantarum, maturation, lanhouin.

Fermentation of wheat: Effects of backslopping different proportions of pre-fermented wheat on the microbialand chemical composition

The objective of the study was to examine effect of backslop on the chemical and microbiological characteristics of fermented wheat (FW). Coarsely ground wheat was mixed with water (1:3 wt/wt) and inoculated with 6 log cfu ml−1 each of an overnight culture of Lactobacillus plantarum and Pediococcus pentosaceus. Four fermentation treatments were conducted in 45 l, closed, PVC containers over 48 hours. Three treatments investigated the benefits of the addition of previously fermented wheat (backslopping, BSL) at different proportions (0.20, 0.33 or 0.42 kg) to freshly prepared wheat. The control treatment contained no addition of BSL. Elimination of coliforms from the FW within 48 h was only achieved through backslopping; where coliform bacteria counts decreased from approximately 6.5 log10 cfu ml−1 to less than 3 log10 cfu ml−1. There was no apparent advantage in increasing the backslop proportion above 0.20. However, the exclusion of coliform bacteria required the pH to remain below 4.0 for at a minimum of 24 h. The results of these studies indicate that fermentation of wheat has the potential to reduce the risk of feed-borne colibacillosis and provides a practical alternative to producers that cannot ferment multiple diets or have limited fermentation capacity.

Expression of the catalase gene katA in starter culture Lactobacillus plantarum TISTR850 tolerates oxidative stress and reduces lipid oxidation in fermented meat product

The catalase gene katA of Lactobacillus sakei SR911 was cloned and expressed in Escherichia coli UM2 and Lactobacillus plantarum TISTR850 under strong lactococcal promoter P59 in E. coli– lactococcus expression vector pIL1020. The L. plantarum TISTR850 is a catalase-deficient strain isolated from local fermented meat product. The recombinant L. plantarum TISTR850 was shown to decompose hydrogen peroxide, and catalase activity approximately three times higher that of natural catalase-producing strain L. sakei SR911. The recombinant protein was also detected by in situ activity staining of the catalase enzyme. The recombinant L. plantarum TISTR850 did not accumulate hydrogen peroxide under glucose-limited aerobic conditions and remained viable after 60 h of incubation. The recombinant and host strain L. plantarum TISTR850 were used as starter cultures in the fermented meat product, and lipid oxidation was monitored over a 7-day storage at 20 °C determined as thiobarbituric acid-reactive substances (TBARS) value. The lipid oxidation level in the fermented meat product seeded with the catalase genetically modified starter culture L. plantarum TISTR850 was significantly lower than that of the natural catalase-deficient strain.

Use of bacteriocin-producing starter cultures of Lactobacillus plantarum and Lactobacillus curvatus in production of ostrich meat salami

Ostrich meat salami was produced by using Lactobacillus plantarum strain 423 and Lactobacillus curvatus strain DF126. The strains produce the bacteriocins plantaricin 423 and curvacin DF126, respectively. The specific activity of plantaricin 423 in MRS broth at 30 °C increased as the pH decreased from 6.5 to 3.5, but activity subsequently decreased. The activity of curvacin DF126 increased under the same conditions, but remained stable for the duration of the growth cycle. Maximum curvacin DF126 and plantaricin 423 activity levels were recorded at a culture pH of around 4. The spectra of antimicrobial activity recorded for plantaricin 423 and curvacin DF126 were similar. Neither of the two bacteriocins inhibited the growth of Micrococcus sp. MC50 and did not have any inhibitory effect on either of the producer strains. Curvacin DF126 and plantaricin 423 inhibited the growth of L. monocytogenes in salami meat. However, after 15 h of fermentation the viable count of L. monocytogenes LM1 increased, probably due to a decrease in activity of the bacteriocins and/or the development of resistant bacterial cells. This is the first report on the inhibition of L. monocytogenes in ostrich meat salami by using bacteriocinogenic starter cultures.

The use of starter cultures to produce ‘Pito’, a Nigerian fermented alcoholic beverage

‘Pito’ a traditional Nigerian alcoholic beverage was brewed in the laboratory with pure cultures of Lactobacillus plantarum in combination with Saccharomyces cerevisiae and Pediococcus halophilus in combination with Candida tropicalis isolated from a local brew. The pH, colour, titrable acidity, alcohol content, specific gravity, taste and flavour of ‘Pito’ produced by this method compared favourably with that produced by the traditional method. The method achieved a reduced fermentation time and an improved production process. It is recommended that Pito be produced with Lactobacillus plantarum in combination with Saccharomyces cerevisiae.