Effect of Cocultivation on Lactobacillus plantarum Strains Growth and Antagonistic Activity

2021 ◽  
Vol 83 (1) ◽  
pp. 12-20
Author(s):  
I.L. Garmasheva ◽  
◽  
O.M. Vasyliuk ◽  
L.T. Oleschenko ◽  
◽  
...  

The use of bacterial starters for the production of fermented foods has several advantages over traditional spontaneous fermentation, as it provides a rapid and controlled decrease of pH, improves the microbiological quality of the product, and prolongs the shelf-life. Fermented foods are typically produced using mixed cultures of lactic acid bacteria (LAB) due to the synergism between their constituent bacterial cultures. So, the compatibility of the LAB strains decides the efficacy of a multi-strain starter. The purpose of this study was to investigate the effect of the cocultivation of Lactobacillus plantarum strains on the growth, acidification, and antagonistic activity to determine suitable strain combinations for fermented vegetable production. Methods. The effect of cocultivation on growth characteristics of four L. plantarum strains was determined in MRS medium and cabbage-based medium with 2.5% NaCl. After 8 h of cultivation at 30°C and 37°C, the number of viable cells (CFU/ml) and the pH of the medium were determined. The antagonistic activity of monocultures of L. plantarum and their six compositions against opportunistic pathogenic microorganisms was determined by the method of delayed antagonism. Results. During growth in MRS broth at 30°C cocultivation of L. plantarum 47SM with L. plantarum 691T or L. plantarum 1047K strains led to enhanced rates of growth compared to the monocultures, suggesting some degree of symbiosis between these strains. Viable cell counts of L. plantarum 47SM, 1047K and 691T strains and ΔpH values of L. plantarum 952K, 1047K, and 691T strains were higher after 8 h growth in the cabbage-based medium at 30°C compared to MRS broth. Despite the intensive growth of L. plantarum monocultures in cabbage-based medium, a significant decrease of viable cell counts and ΔpH values during cocultivation at 30°C were found. Cocultivation did not affect the average size of the growth inhibition zones of most of the indicator strains used. However, growth inhibition zones of Shigella flexneri, Escherichia coli, and Proteus vulgaris decreased in some L. plantarum mixed cultures compared to monocultures. Thus, the growth inhibition zones of E. coli and S. flexneri by mixed culture L. plantarum 47 SM+1047K were significantly smaller compared to the growth inhibition zones of L. plantarum monocultures. Conclusions. Thus, based on the data obtained in present work, we can assume that some of these L. plantarum strains used in the work may be bactericinogenic. Although the four L. plantarum strains studied are compatible when cocultivated in a standard rich MRS medium, the results of cocultivation in a cabbage-based medium with 2.5% NaCl does not allow to recommend the use of these L. plantarum strains simultaneously in the starter for vegetable fermentation. Further investigation of bacteriocinogenic properties and mechanisms of growth inhibition under cocultivation in vegetable-like conditions are needed, which will allow combining of some of these L. plantarum strains with LAB strains of other species or genera to create multi-starters for vegetable fermentation.

2019 ◽  
Vol 1 (1) ◽  
Author(s):  
Ronamae Bradford ◽  
Vondel Reyes ◽  
Franklin Bonilla ◽  
Federico Bueno ◽  
Bennett Dzandu ◽  
...  

Abstract The objectives of this study were to develop a probiotic milk powder containing Lactobacillus plantarum NCIMB 8826 immobilized with prebiotic Hi-maize starch and to analyze cell viability after spray drying and exposure to simulated gastric and intestinal conditions. Milk powders containing free L. plantarum and cells immobilized with Hi-maize starch were assessed. Powders were evaluated during storage at 4 °C for 15 days. After spray drying, at 0 and 15 days of storage both treatments had over 8 log CFU/g of viable cells and there were higher viable counts found for immobilized cells compared to free cells after 120 min in simulated gastric fluid. At 15 days of storage, immobilized cells had higher viable counts than free cells after exposure to simulated intestinal fluid for 120 min. The combined probiotic and prebiotic milk powder had stable viable cell counts at refrigerated storage conditions and under simulated gastric and intestinal transit. Graphical abstract


2002 ◽  
Vol 68 (9) ◽  
pp. 4399-4406 ◽  
Author(s):  
Adriana Molina-Gutierrez ◽  
Volker Stippl ◽  
Antonio Delgado ◽  
Michael G. Gänzle ◽  
Rudi F. Vogel

ABSTRACT Hydrostatic pressure may affect the intracellular pH of microorganisms by (i) enhancing the dissociation of weak organic acids and (ii) increasing the permeability of the cytoplasmic membrane and inactivation of enzymes required for pH homeostasis. The internal pHs of Lactococcus lactis and Lactobacillus plantarum during and after pressure treatment at 200 and 300 MPa and at pH values ranging from 4.0 to 6.5 were determined. Pressure treatment at 200 MPa for up to 20 min did not reduce the viability of either strain at pH 6.5. Pressure treatment at pH 6.5 and 300 MPa reduced viable cell counts of Lactococcus lactis and Lactobacillus plantarum by 5 log after 20 and 120 min, respectively. Pressure inactivation was faster at pH 5 or 4. At ambient pressure, both strains maintained a transmembrane pH gradient of 1 pH unit at neutral pH and about 2 pH units at pH 4.0. During pressure treatment at 200 and 300 MPa, the internal pH of L. lactis was decreased to the value of the extracellular pH during compression. The same result was observed during treatment of Lactobacillus plantarum at 300 MPa. Lactobacillus plantarum was unable to restore the internal pH after a compression-decompression cycle at 300 MPa and pH 6.5. Lactococcus lactis lost the ability to restore its internal pH after 20 and 4 min of pressure treatment at 200 and 300 MPa, respectively. As a consequence, pressure-mediated stress reactions and cell death may be considered secondary effects promoted by pH and other environmental conditions.


Author(s):  
Febry Surbakti

Di Indonesia terkenal dengan berbagai macam makanan tradisional fermentasi. Jenis makanan tradisional yang banyak dijumpai yang diproses dengan menggunakan proses fermentasi adalah tape, asinan, acar dan lainnya yang mengandung banyak bakteri asam laktat (BAL). Bakteri asam laktat ini merupakan kekayaan alam mikroba yang masih harus dieksplorasi dibidang kesehatannya. Penelitian ini bertujuan untuk mengetahui jenis-jenis dan karakteristik bakteri asam laktat yang terdapat pada acar ketimun. Penelitian ini menggunakan metode deskriptif. Teknik pengambilan sampel langsung dilakukan secara aseptis. Bahan yang digunakan adalah bakteri asam laktat yang diambil dari cairan acar ketimun dengan variasi isi ketimun, ketimun nenas, ketimun nenas dan cabai dan ditumbuhkan pada media MRS Broth (de Man Rogosa Sharpe). Berdasarkan hasil penelitian identifikasi dan karakterisasi bakteri asam laktat pada acar ketimun dengan menggunakan uji morfologi, pewarnaan gram dan uji biokimia maka diperoleh 2 jenis bakteri asam laktat pada acar ketimun. Adapun bakteri asam laktat yang ditemukan antara lain adalah Lactobacillus plantarum dan Lactobacillus reuteri. Karakterisitik secara umum adalah basil, gram positif, katalase negatif, tidak motil, suhu optimum 250°C- 300°C dan memiliki tipe fermentasi heterofermentatif dan homofermentatif.  ABSTRACT: In Indonesia it is famous for various kinds of traditional fermented foods. Traditional food types that are often found that are processed using fermentation are tape, pickled, pickled and others that contain a lot of lactic acid bacteria (LAB). This lactic acid bacteria is a natural wealth of microbes that must still be explored in the field of health. This study aims to determine the types and characteristics of lactic acid bacteria found in pickles to Cucumber. This research uses a descriptive method. The direct sampling technique is carried out aseptically. The material used is lactic acid bacteria taken from pickled liquid to Cucumber with various contents to Cucumber, to Pineapple Cucumber, to Cucumber pineapple and chili and grown on MRS Broth media (de Man Rogosa Sharpe). Based on the results of the study of identification and characterization of lactic acid bacteria on pickles to Cucumber using morphological tests, gram staining and biochemical tests, 2 types of lactic acid bacteria were obtained on pickles to Cucumber. The lactic acid bacteria found include Lactobacillus plantarum and Lactobacillus reuteri. General characteristics are basil, gram positive, negative catalase, not motile, optimum temperature 250C-300C and have heterofermentative and homofermentative fermentation types. Keywords: Lactic acid bacteria, Probiotik, Pickled cucumber


2017 ◽  
Vol 36 (4) ◽  
pp. 424
Author(s):  
Neti Yuliana ◽  
Tintan Noviyeziana ◽  
Sutikno Sutikno

The objective of this study was to characterize the durian lay juices supplemented by Lactic Acid Bacteria (LAB)  culture during cold storage. The LAB cultures consisted of Lactobacillus plantarum FNCC-0265, Lactobacillus acidophillus FNCC-0051 and combination of these LABs. After being inoculated with a 24 h – old culture starters, durian lay juice was incubated at 37 °C for 2 hours and kept at 4 °C for 0, 1, 2, 3 and 4 weeks. Changes in pH, total lactic acid, and sugar content, viable cell counts of LAB, total soluble solid, viability of LAB, and sensory characteristic during fermentation were measured. The results revealed that pH decreased from 4.79 to 4.20; sugar content reduced from 4.52 to 0.75 %; total soluble solid reduced from 8.93 to 7.97 °Bx, and sensory score reduced from 3.71 (like) to 3.40 (moderate like); while  the total lactic acid increased from 0.33 to 0.69 %; viable cell counts of LAB increased from 8.86  to 9.14 log CFU/ml; and % viability of LAB decreased from 100 to 95.90, during storage. Combination of Lactobacillus plantarum FNCC-0265 and Lactobacillus acidophillus FNCC-0051 cultures at two weeks cold storage period was the best treatment, based on the sensory and viable cell counts of LAB. The characteristics of this durian lay juice probiotic were pH 4.46, total lactic acid 0.59 %, sugar content 2.13 %, viable cell counts of LAB 9.33 log CFU/mL, total soluble solid 9.60 °Bx, and carbohydrate 45,05 %. This durian lay juice contained oxalate (107.56 mg/L), malic (527.895 mg/L), lactic (4381.49 mg/L) and citric (351.245 mg/L) acid.ABSTRAKTujuan penelitian ini adalah mengkaji karakteristik minuman laktat sari durian lay yang disuplementasi dengan kultur bakteri asam laktat (BAL) selama penyimpanan pada suhu rendah. Kultur BAL terdiri dari Lactobacillus plantarum FNCC-0265, Lactobacillus acidophillus FNCC-0051, dan campuran keduanya.  Sari buah durian lay diinokulasi dengan kultur berumur 24 jam lalu diinkubasi pada suhu 37 °C selama 2 jam dan kemudian disimpan selama 0, 1, 2, 3, dan 4 minggu. Perubahan pH, total asam, gula, total sel BAL. Total padatan terlarut, viabilitas BAL, dan sensori selama penyimpanan dingin diukur. Hasil penelitian menunjukkan bahwa terjadi perubahan pH dari 4,79 ke 4,20; kandungan gula dari 4,52 ke 0,75%; total padatan terlarut dari 8,93 ke 7.97 °Bx, viabilitas BAL dari 100 % menjadi 95,90 %,  dan skor sensori dari 3,71 suka ke 3,40 (agak suka); sementara pada total asam terjadi peningkatan dari 0,33 ke 0,69 %; dan sel hidup BAL meningkat dari 8,86 menjadi 9,14 log CFU/mL. Berdasarkan parameter sensori, perlakuan kultur campuran Lactobacillus plantarum FNCC-0265 dan Lactobacillus acidophillus FNCC-0051 yang disimpan selama 2 minggu merupakan perlakuan terbaik. Sari buah durian lay ini mempunyai karakteristik sebagai berikut: pH 4,45, total asam laktat 0,83 %, gula 2,20%, sel hidup BAL 9,33 log CFU/mL, total padatan terlarut  9,16 °Bx, dan karbohidrat 35,04 %. Asam organik yang terkandung pada jus durian lay ini adalah asam oksalat (108 mg/L), malat (528 mg/L), laktat (4382 mg/L) dan sitrat (351 mg/L).


2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S643-S643
Author(s):  
Maria F Mojica ◽  
Christopher Bethel ◽  
Emilia Caselli ◽  
Magdalena A Taracila ◽  
Fabio Prati ◽  
...  

Abstract Background Catalytic mechanisms of serine β-lactamases (SBL; classes A, C and D) and metallo-β-lactamases (MBLs) have directed divergent strategies towards inhibitor design. SBL inhibitors act as high affinity substrates that -as in BATSIs- form a reversible, dative covalent bond with the conserved active site Ser. MBL inhibitors bind the active-site Zn2+ ions and displace the nucleophilic OH-. Herein, we explore the efficacy of a series of BATSI compounds with a free-thiol group at inhibiting both SBL and MBL. Methods Exploratory compounds were synthesized using stereoselective homologation of (+) pinandiol boronates to introduce the amino group on the boron-bearing carbon atom, which was subsequently acylated with mercaptopropanoic acid. Representative SBL (KPC-2, ADC-7, PDC-3 and OXA-23) and MBL (IMP-1, NDM-1 and VIM-2) were purified and used for the kinetic characterization of the BATSIs. In vitro activity was evaluated by a modified time-kill curve assay, using SBL and MBL-producing strains. Results Kinetic assays revealed that IC50 values ranged from 1.3 µM to >100 µM for this series. The best compound, s08033, demonstrated inhibitory activity against KPC-2, VIM-2, ADC-7 and PDC-3, with IC50 in the low μM range. Reduction of at least 1.5 log10-fold of viable cell counts upon exposure to sub-lethal concentrations of antibiotics (AB) + s08033, compared to the cells exposed to AB alone, demonstrated the microbiological activity of this novel compound against SBL- and MBL-producing E. coli (Table 1). Table 1 Conclusion Addition of a free-thiol group to the BATSI scaffold increases the range of these compounds resulting in a broad-spectrum inhibitor toward clinically important carbapenemases and cephalosporinases. Disclosures Robert A. Bonomo, MD, Entasis, Merck, Venatorx (Research Grant or Support)


2006 ◽  
Vol 72 (5) ◽  
pp. 3482-3488 ◽  
Author(s):  
M�nica Ordax ◽  
Ester Marco-Noales ◽  
Mar�a M. L�pez ◽  
Elena G. Biosca

ABSTRACT Copper compounds, widely used to control plant-pathogenic bacteria, have traditionally been employed against fire blight, caused by Erwinia amylovora. However, recent studies have shown that some phytopathogenic bacteria enter into the viable-but-nonculturable (VBNC) state in the presence of copper. To determine whether copper kills E. amylovora or induces the VBNC state, a mineral medium without copper or supplemented with 0.005, 0.01, or 0.05 mM Cu2+ was inoculated with 107 CFU/ml of this bacterium and monitored over 9 months. Total and viable cell counts were determined by epifluorescence microscopy using the LIVE/DEAD kit and by flow cytometry with 5-cyano-2,3-ditolyl tetrazolium chloride and SYTO 13. Culturable cells were counted on King's B nonselective solid medium. Changes in the bacterial morphology in the presence of copper were observed by scanning electron microscopy. E. amylovora entered into the VBNC state at all three copper concentrations assayed, much faster when the copper concentration increased. The addition of different agents which complex copper allowed the resuscitation (restoration of culturability) of copper-induced VBNC cells. Finally, copper-induced VBNC cells were virulent only for the first 5 days, while resuscitated cells always regained their pathogenicity on immature fruits over 9 months. These results have shown, for the first time, the induction of the VBNC state in E. amylovora as a survival strategy against copper.


2015 ◽  
Vol 12 (106) ◽  
pp. 20150069 ◽  
Author(s):  
Hiroki Takahashi ◽  
Taku Oshima ◽  
Jon L. Hobman ◽  
Neil Doherty ◽  
Selina R. Clayton ◽  
...  

Zinc is essential for life, but toxic in excess. Thus all cells must control their internal zinc concentration. We used a systems approach, alternating rounds of experiments and models, to further elucidate the zinc control systems in Escherichia coli . We measured the response to zinc of the main specific zinc import and export systems in the wild-type, and a series of deletion mutant strains. We interpreted these data with a detailed mathematical model and Bayesian model fitting routines. There are three key findings: first, that alternate, non-inducible importers and exporters are important. Second, that an internal zinc reservoir is essential for maintaining the internal zinc concentration. Third, our data fitting led us to propose that the cells mount a heterogeneous response to zinc: some respond effectively, while others die or stop growing. In a further round of experiments, we demonstrated lower viable cell counts in the mutant strain tested exposed to excess zinc, consistent with this hypothesis. A stochastic model simulation demonstrated considerable fluctuations in the cellular levels of the ZntA exporter protein, reinforcing this proposal. We hypothesize that maintaining population heterogeneity could be a bet-hedging response allowing a population of cells to survive in varied and fluctuating environments.


Author(s):  
Yash S. Raval ◽  
Abdelrhman Mohamed ◽  
Jayawant N. Mandrekar ◽  
Cody Fisher ◽  
Kerryl E. Greenwood-Quaintance ◽  
...  

Wound infections are caused by bacteria and/or fungi. The presence of fungal biofilms in wound beds presents a unique challenge, as fungal biofilms may be difficult to eradicate. The goal of this work was to assess the in vitro anti-biofilm activity of a H 2 O 2 -producing electrochemical bandage (e-bandage) against 15 yeast isolates representing commonly-encountered species. Time-dependent decreases in viable biofilm CFU counts of all isolates tested were observed, resulting in no visible colonies with 48 hours of exposure by plate culture. Fluorescence microscopic analysis showed extensive cell membrane damage of biofilm cells after e-bandage treatment. Reductions in intracellular ATP levels of yeast biofilm cells were recorded post e-bandage treatment. Our results suggest that exposure to H 2 O 2 -producing e-bandages reduce in vitro viable cell counts of yeast biofilms, making this a potential new topical treatment approach for fungal wound infections.


1991 ◽  
Vol 54 (5) ◽  
pp. 349-353 ◽  
Author(s):  
AMECHI OKEREKE ◽  
THOMAS J. MONTVILLE

Twenty-three strains of lactic acid bacteria were tested by deferred antagonism methods for bacteriocin-like activity against types A and B spores from 11 proteolytic and nonproteolytic Clostridium botulinum strains. Pediococcus pentosaceus ATCC 43200, Pediococcus pentosaceus ATCC 43201, Lactococcus lactis subsp. lactis ATCC 11454, Lactobacillus acidophilus N2, Lactobacillus plantarum Lb75, Lactobacillus plantarum Lb592, and Lactobacillus plantarum BN exhibited bacteriocin-like inhibition of all C. botulinum strains tested. By excluding inhibition due to hydrogen peroxide, acid, and lytic phage and confirming their proteinaceous nature, the inhibitors were confirmed as bacteriocins. The minimum inhibitory cell concentrations (MICC) required to produce 1 mm radius inhibition zones were determined by direct antagonism testing. Only strains 43200, 43201, 11454, and N2 were inhibitory when cultured simultaneously with the botulinal spores. The MICCs of strains antagonistic to C. botulinum spores by simultaneous testing ranged between 1.6 × 105and 4.7 × 107CFU/ml. Based on the MICCs, P. pentosaceus 43200 was most inhibitory to C. botulinum.


2018 ◽  
Vol 15 (1) ◽  
pp. 1
Author(s):  
Fathyah Hanum Pamungkaningtyas ◽  
Mariyatun Mariyatun ◽  
Rafli Zulfa Kamil ◽  
Ryan Haryo Setyawan ◽  
Pratama Nur Hasan ◽  
...  

Lactic acid bacteria have been isolated from several Indonesian indigenous fermented foods and screened for the potential strains as probiotic candidates. The aim of this study was to evaluate sensory properties and respondents’ preference of yogurt-like set and yogurt-like drink with various Indonesian indigenous probiotic strains produced by dairy industry. Indigenous probiotics of Lactobacillus plantarum MUT-7 and Lactobacillus plantarum DAD-13 were used to produce yogurt-like set and yogurt-like drink. Family perception toward yogurt-like drink was performed in Yogyakarta involving 100 family members. The yogurt-like products were also compared to yogurt containing commercial Lactobacilus bulgaricus and Streptococus thermophilus or commercial yogurt produced by dairy company. Several sensory evaluation toward sensory properties and panelist’s preference were performed in different cities.  The result showed that the indigenous probiotic L. plantarum DAD-13 and L. plantarum MUT-7 were potential to be used as a starter culture for the production of yogurt-like set and yogurt-like drink. The combination of indigenous probiotics and indigenous lactic acid bacteria S. thermophilus DAD-11 resulted in better sensory properties of yogurt set compared to combination of L. bulgaricus and S. thermophilus. 


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