Сравнительное изучение антагонистической активности бифидо- и лактосодержащих пробиотиков

2021 ◽  
Author(s):  
В. Несчисляев ◽  
М. Столбова ◽  
Е. Орлова ◽  
А. Савина ◽  
И. Белова
Keyword(s):  
Escherichia Coli ◽  
Lactobacillus Plantarum ◽  
Enterococcus Faecium ◽  
Bifidobacterium Bifidum ◽  
E Coli

Проведен сравнительный анализ антагонистической активности пробиотических препаратов: монокомпонентных — Бифидумбактерин (Bifidobacterium bifidum), Лактобактерин (Lactobacillus plantarum); поликомпонентных пробиотиков — Бифилакт-БИЛС (L. plantarum, L. acidophilus, B. bifidum), Линекс (L. acidophilus, B. infantis, Enterococcus faecium), Бифиформ (B. longum, E. faecium); на основе сорбированных клеток — Бифидумбактерин форте (B. bifidum, сорбированный на активированном угле) и Имбикапс (B. bifidum, сорбированный на гомогенате бурых водорослей) — методом отсроченного антагонизма (в качестве тест-штамма использовали Escherichia coli 157) и экспресс-теста подавления биолюминесценции индикаторного штамма E. coli lum+. Выявлена высокая чувствительность тест-штаммов E. coli 157 и E. coli lum+ к препаратам Лактобактерин, Бифилакт-БИЛС и Имбикапс. Установлено, что экспресс-тест изменения биолюминесценции штамма E. coli lum+ позволяет оценить антагонистическую активность пробиотического препарата в целом, тогда как метод отсроченного антагонизма дает представление о потенциальных антагонистических свойствах штаммов, входящих в состав препаратов, которую они проявляют в благоприятных условиях. В экспресс-тесте также оценивается метаболитная составляющая препарата, которая играет существенную роль в реализации биологической активности пробиотика. Препараты Лактобактерин, Бифилакт-БИЛС и Имбикапс обладают определенным преимуществом по биологической активности в сравнении с аналогичными отечественными и зарубежными пробиотиками.

scholarly journals Caracterización de microorganismos con potencial probiótico aislados de estiércol de terneros Brahman en Sucre, Colombia

2018 ◽  
Vol 29 (2) ◽  
pp. 438
Author(s):  
Pedro Luis Castillo Arroyo ◽  
César Augusto Betancur Hurtado ◽  
Enrique Pardo Pérez
Keyword(s):  
Escherichia Coli ◽  
Enterococcus Faecium ◽  
Candida Krusei ◽  
E Coli

El objetivo del estudio fue caracterizar microorganismos con potencial probiótico de estiércol de terneros lactantes Brahman en Sucre (Colombia). Se aislaron bacterias y levaduras de las muestras de estiércol y se determinó la capacidad probiótica de estas cepas mediante pruebas de resistencia a sales biliares (0.05, 0.1, 0.15 y 0.3%), resistencia a pH ácido (pH 3, 4, 5.6, 7), tolerancia a NaCl (2, 4, 6, 8, 10%) y actividad antagónica (Salmonella sp y Escherichia coli). Nueve microorganismos fueron identificados y tres pasaron las pruebas de tolerancia. Se determinó la capacidad antagónica frente a bacterias patógenas (Salmonella sp y E. coli), evidenciada por halos (mm) de las tres cepas seleccionadas. La identificación de las cepas se realizó por los métodos bioquímicos API 50 CHL V5.1 para Lactobacillus y API 20 AUX para levaduras. El análisis molecular de dos cepas (M13a y 103M2) identificó la cepa M13a como Enterococcus faecium y la cepa 103M2 como Candida krusei. En conclusión, Enterococcus faecium se convierte en una alternativa viable para la formulación de un biopreparado para mejorar los parámetros productivos y disminuir los trastornos gastrointestinales en los terneros lactantes.

scholarly journals Antimicrobial activity of lactic acid bacteria isolated from garri on Escherichia coli strains isolated from clinical and environmental samples

2020 ◽  
Vol 12 (4) ◽  
pp. 357-365
Author(s):  
H.I. Atta ◽  
A. Gimba ◽  
T. Bamgbose
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactobacillus Acidophilus ◽  
Plant Pathogens ◽  
Resistant Bacteria ◽  
Fermented Foods ◽  
Environmental Isolates ◽  
E Coli

Abstract. The production of bacteriocins by lactic acid bacteria affords them the ability to inhibit the growth of bacteria; they are particularly important in the biocontrol of human and plant pathogens. Lactic acid bacteria have been frequently isolated from fermented foods due to the high acidity these foods contain. In this study, lactic acid bacteria were isolated from garri, a popular Nigerian staple food, which is fermented from cassava, and their antagonistic activity against clinical and environmental isolates of Escherichia coli was determined. The species of Lactobacillus isolated include: Lactobacillus plantarum (50%), Lactobacillus fermentum (20%), Lactobacillus acidophilus (20%), and Lactobacillus salivarius (10%). Growth inhibition of the strains of E.coli was observed in Lactobacillus plantarum that inhibited the growth of both. The clinical and environmental isolates of E. coli were inhibited by Lactobacillus plantarum, while Lactobacillus acidophilus showed activity against only the clinical isolate. The greatest zone of inhibition against the strains of E. coli was recorded by Lactobacillus acidophilus (22.7±1.53 mm). The bacteriocins produced by Lactobacillus species have a good potential in the biocontrol of pathogens, and should be the focus of further studies on antibiotic resistant bacteria.

scholarly journals CONSTRUCTION, EXPRESSION AND PURIFICATION OF RECOMBINANT PRE-MATURE PEPTIDE OF PLANTARICIN F FROM Lactobacillus plantarum S34 IN Escherichia coli

2015 ◽  
Vol 16 (1) ◽  
pp. 31
Author(s):  
Kusdianawati Kusdianawati ◽  
Apon Zaenal Mustopa ◽  
Suharsono Suharsono ◽  
Bugi Ratno Budiarto ◽  
Fatimah Fatimah ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactobacillus Plantarum ◽  
Proteolytic Enzymes ◽  
Leader Peptide ◽  
Human Consumption ◽  
Mature Peptide ◽  
Expression And Purification ◽  
Food Preservative ◽  
E Coli ◽  
Sds Page

Plantaricin is one of bacteriocins that have the potential to be used as food preservative. Plantaricin is safe for human consumption because it can be easily degraded by proteolytic enzymes. The objective of this study was to express and purify recombinant pre-mature peptide of plantaricin F from <em>Lactobacillus plantarum</em> S34 in <em>Escherichia coli</em>. Plantaricin gene-specific primer was used to obtain pln F structural gene amplicon from L. <em>plantarum</em> S34. This amplicon was cloned in pET32a vector and expressed in E. coli BL21 (DE3) pLysS. Pre-mature plantaricin F peptide was expressed as Histagged-fusion protein and separated by Co2+-chelating affinity chromatography. L. <em>plantarum</em> S34-derived pre-mature plantaricin F peptide fused with thioredoxin-(His)6tag had successfully been expressed in E. <em>coli</em> BL21 (DE3) pLysS using pET32a as an expression vector. The fused recombinant pln F as pre-mature state expressed had a molecular mass of +24 kDa, meanwhile the fused recombinant that contained only the leader peptide of pln F appeared as +20 kDa based on SDS-PAGE separations. The optimal production of fused recombinant pln F as soluble fraction was obtained when culture condition was added with 0.5 mM of IPTG and incubated at 22°C for 5 hours (OD~1). Furthermore, the expression of fused recombinant pln F as its pre-mature peptide pointed out that the pln F’s leader peptide could be proteolytically cleaved by a system in heterologous cells. Overall, heterologous pln F production as pre-mature peptide fused with thioredoxin-(His)6tag had been well established. From this research, we expect plantaricin F can be expressed and purified in E. coli.

scholarly journals Retrospective Analysis on Antimicrobial Resistance Trends and Prevalence of β-lactamases in Escherichia coli and ESKAPE Pathogens Isolated from Arabian Patients during 2000–2020

2020 ◽  
Vol 8 (10) ◽  
pp. 1626
Author(s):  
Mahfouz Nasser ◽  
Snehal Palwe ◽  
Ram Naresh Bhargava ◽  
Marc G. J. Feuilloley ◽  
Arun S. Kharat
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Enterococcus Faecium ◽  
Arab Countries ◽  
Clinical Isolates ◽  
Arab Region ◽  
E Coli ◽  
Extended Spectrum ◽  
Eskape Pathogens

The production of diverse and extended spectrum β-lactamases among Escherichia coli and ESKAPE pathogens is a growing threat to clinicians and public health. We aim to provide a comprehensive analysis of evolving trends of antimicrobial resistance and β-lactamases among E. coli and ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acine to bacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) in the Arabian region. A systematic review was conducted in Medline PubMed on papers published between January 2000 and February 2020 on countries in the Arab region showing different antibiotic resistance among E. coli and ESKAPE pathogens. A total of n = 119,144 clinical isolates were evaluated for antimicrobial resistance in 19 Arab countries. Among these clinical isolates, 74,039 belonged to E. coli and ESKAPE pathogen. Distribution of antibiotic resistance among E. coli and ESKAPE pathogens indicated that E. coli (n = 32,038) was the predominant pathogen followed by K. pneumoniae (n = 17,128), P. aeruginosa (n = 11,074), methicillin-resistant S. aureus (MRSA, n = 4370), A. baumannii (n = 3485) and Enterobacter spp. (n = 1574). There were no reports demonstrating Enterococcus faecium producing β-lactamase. Analyses revealed 19 out of 22 countries reported occurrence of ESBL (Extended-Spectrum β-Lactamase) producing E. coli and ESKAPE pathogens. The present study showed significantly increased resistance rates to various antimicrobial agents over the last 20 years; for instance, cephalosporin resistance increased from 37 to 89.5%, fluoroquinolones from 46.8 to 70.3%, aminoglycosides from 40.2 to 64.4%, mono-bactams from 30.6 to 73.6% and carbapenems from 30.5 to 64.4%. An average of 36.9% of the total isolates were reported to have ESBL phenotype during 2000 to 2020. Molecular analyses showed that among ESBLs and Class A and Class D β-lactamases, blaCTX-M and blaOXA have higher prevalence rates of 57% and 52.7%, respectively. Among Class B β-lactamases, few incidences of blaVIM 27.7% and blaNDM 26.3% were encountered in the Arab region. Conclusion: This review highlights a significant increase in resistance to various classes of antibiotics, including cephalosporins, β-lactam and β-lactamase inhibitor combinations, carbapenems, aminoglycosides and quinolones among E. coli and ESKAPE pathogens in the Arab region.

scholarly journals Cloning and Heterologous Expression of Plantaricin ZJ5, a Novel Bacteriocin from Lactobacillus plantarum ZJ5, in Escherichia coli

2018 ◽  
Vol 13 (12) ◽  
pp. 1934578X1801301
Author(s):  
Dafeng Song ◽  
Ping Li ◽  
Qing Gu
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Lactobacillus Plantarum ◽  
Micrococcus Luteus ◽  
Biologically Active ◽  
Biological Method ◽  
Lac Operon ◽  
E Coli ◽  
First Time ◽  
Soluble Material

A novel bacteriocin, plantaricin ZJ5 (PZJ5) was yielded from Lactobacillus plantarum ZJ5, cloned, and produced in Escherichia coli BL21 (DE3) pLys. The PZJ5 structural gene was fused with a Trx tag, and cloned into the pET32a plasmid under the control of the inducible lac operon. Induction was performed with isopropyl-β-D-thiogalactopyranoside (IPTG), with subsequent overexpression of the fusion protein, followed by purification to homogeneity via His affinity chromatography. Recombinant E. coli produced greater quantities of PZJ5 than L. plantarum ZJ5, and PZJ5 in E. coli was expressed in the form of soluble material. Biologically active PZJ5 was recovered by cleaving the purified fusion protein using enterokinase. The released PZJ5 demonstrated antibacterial activity against Micrococcus luteus. In this study, an inexpensive biological method using a Trx fusion system was presented, and for the first time, bacteriocin PZJ5 was expressed and purified in E. coli.

scholarly journals In Vitro Activity of Trovafloxacin against Bacteroides fragilis in Mixed Culture with either Escherichia coli or a Vancomycin- Resistant Strain of Enterococcus faecium Determined by an Anaerobic Time-Kill Technique

2001 ◽  
Vol 45 (1) ◽  
pp. 243-251 ◽  
Author(s):  
Lorna E. T. Stearne ◽  
Clarissa Kooi ◽  
Wil H. F. Goessens ◽  
Irma A. J. M. Bakker-Woudenberg ◽  
Inge C. Gyssens
Keyword(s):  
Escherichia Coli ◽  
Resistant Strain ◽  
Enterococcus Faecium ◽  
Mixed Cultures ◽  
E Coli ◽  
Abdominal Abscesses ◽  
Pure Cultures ◽  
Vancomycin Resistant ◽  
Time Kill

ABSTRACT To determine the efficacy of trovafloxacin as a possible treatment for intra-abdominal abscesses, we have developed an anaerobic time-kill technique using different inocula to study the in vitro killing ofBacteroides fragilis in pure culture or in mixed culture with either Escherichia coli or a vancomycin-resistant strain of Enterococcus faecium (VREF). With inocula of 5 × 105 CFU/ml and trovafloxacin concentrations of ≤2 μg/ml, a maximum observed effect (E max) of ≥6.1 (log10 CFU/ml) was attained with all pure and mixed cultures within 24 h. With inocula of 108CFU/ml, a similar E max and a similar concentration to produce 50% of E max(EC50) for B. fragilis were found in both pure cultures and mixed cultures with E. coli. However, to produce a similar killing of B. fragilis in the mixed cultures with VREF, a 14-fold increase in the concentration of trovafloxacin was required. A vancomycin-susceptible strain of E. faecium and a trovafloxacin-resistant strain of E. coli were also found to confer a similar “protective” effect on B. fragilis against the activity of trovafloxacin. Using inocula of 109 CFU/ml, the activity of trovafloxacin was retained for E. coli and B. fragilis and was negligible against VREF. We conclude that this is a useful technique to study the anaerobic killing of mixed cultures in vitro and may be of value in predicting the killing of mixed infections in vivo. The importance of using mixed cultures and not pure cultures is clearly shown by the difference in the killing of B. fragilis in the mixed cultures tested. Trovafloxacin will probably be ineffective in the treatment of infections involving large numbers of enterococci. However, due to its ability to retain activity against large cultures of B. fragilis and E. coli, trovafloxacin could be beneficial in the treatment of intra-abdominal abscesses.

scholarly journals Inactivation of PadR, the Repressor of the Phenolic Acid Stress Response, by Molecular Interaction with Usp1, a Universal Stress Protein from Lactobacillus plantarum, in Escherichia coli

2009 ◽  
Vol 75 (16) ◽  
pp. 5273-5283 ◽  
Author(s):  
Jérôme Gury ◽  
Hélène Seraut ◽  
Ngoc Phuong Tran ◽  
Lise Barthelmebs ◽  
Stéphanie Weidmann ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Lactobacillus Plantarum ◽  
Phenolic Acid ◽  
Stress Protein ◽  
Acid Stress ◽  
Universal Stress Protein ◽  
E Coli ◽  
Gram Positive Bacteria ◽  
Acid Stress Response

ABSTRACT The phenolic acid decarboxylase gene padA is involved in the phenolic acid stress response (PASR) in gram-positive bacteria. In Lactobacillus plantarum, the padR gene encodes the negative transcriptional regulator of padA and is cotranscribed with a downstream gene, usp1, which encodes a putative universal stress protein (USP), Usp1, of unknown function. The usp1 gene is overexpressed during the PASR. However, the role and the mechanism of action of the USPs are unknown in gram-positive bacteria. Therefore, to gain insights into the role of USPs in the PASR; (i) a usp1 deletion mutant was constructed; (ii) the two genes padR and usp1 were coexpressed with padA under its own promoter as a reporter gene in Escherichia coli; and (iii) molecular in vitro interactions between the PadR, Usp1, and the padA promoter were studied. Although the usp1 mutant strain retained phenolic acid-dependent PAD activity, it displayed a greater sensitivity to strong acidic conditions compared to that of the wild-type strain. PadR cannot be inactivated directly by phenolic acid in E. coli recombinant cultures but is inactivated by Usp1 when the two proteins are coexpressed in E. coli. The PadR inactivation observed in recombinant E. coli cells was supported by electrophoretic mobility shift assays. Although Usp1 seems not to be absolutely required for the PASR, its capacity to inactivate PadR indicates that it could serve as an important mediator in acid stress response mechanisms through its capacity to interact with transcriptional regulators.

scholarly journals Caracterización molecular de bacterias con potencial probiótico aisladas de heces de neonatos humanos

2019 ◽  
Vol 26 (1) ◽  
pp. 119-130
Author(s):  
Ricardo Santos ◽  
Elizabeth Paitán ◽  
Alejandrina Sotelo ◽  
Doris Zúñiga ◽  
Carlos Vílchez
Keyword(s):  
Escherichia Coli ◽  
16S Rrna ◽  
Enterococcus Faecium ◽  
Salmonella Enterica ◽  
E Coli

El objetivo de este estudio es caracterizar molecularmente bacterias con potencial probiótico aisladas de heces de neonatos humanos. Se evaluó 60 muestras de heces de neonatos (0-3 días) se enriquecieron en caldo Man Rogosa y Sharp (MRS) a 37°C/24h. Se seleccionó y se sometió a pruebas in vitro con sales biliares, resistencia a pH bajo y actividad antimicrobiana frente a Escherichia coli ATCC25922, E. coli ATCC35218, Salmonella enterica y Listeria inocua mediante el ensayo difusión en agar. La identificación molecular se realizó con amplificaciones PCR-BOX y el secuenciamiento del gen 16S rRNA. Se aislaron un total de 48 cepas y todas presentaron resistencia a pH 3 y 0.3% sales biliares; 3 cepas mostraron actividad antimicrobiana frente a E. coli ATCC25922, 1 cepa frente a E. coli ATCC35218, 5 cepas frente a L. inocua y todas frente a S. entérica. De las 48 cepas se obtuvieron dos perfiles BOX-PCR pertenecientes a los géneros de Lactobacillus y Enterococcus. Nueve cepas (C52, C61, C71, C112, C16 2, C192, C20, C35, y C42) presentaron un 100% de similaridad a L. plantarum ATCC 14917T [ACGZ01000098] y dos cepas (C15 y C40) un 99.93% y 99.80% de similaridad, respectivamente a Enterococcus faecium CGMCC 1.2136T [AJKH01000109]; estas cepas mostraron actividad en leche con diferencias significativas (p valor < 0.05) en la cinética de pH 3. En conclusión se encontró bacterias con potencial probiótico.

scholarly journals Inhibitory Potential Of Lactobacillus Plantarum on Escherichia Coli

2017 ◽  
Vol 74 (2) ◽  
pp. 99 ◽  
Author(s):  
Laura MITREA ◽  
Lavinia Florina CĂLINOIU ◽  
Gabriela PRECUP ◽  
Maria BINDEA ◽  
Bogdan RUSU ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactobacillus Plantarum ◽  
Alternative Therapy ◽  
Optimal Parameters ◽  
E Coli ◽  
Enteropathogenic Bacteria ◽  
Probiotic Strains ◽  
Inhibitory Potential ◽  
Mrs Broth

An alternative therapy to diarrhoeal episodes induced by E. coli is represented by probiotic strains (e.g Lactobacillus), which are able to maintain the gut micro-flora in optimal parameters. This work highlights the inhibitory potential of probiotics (L. plantarum ATCC 8014) over the enteropathogenic bacteria like E. coli ATCC 25922. For this study we co-incubated L. plantarum with E. coli in MRS broth at different concentrations (109, 108 CFU/mL – L. plantarum with 105, 104 CFU/mL – E. coli, 4 groups) for 24, 48 and 72h at 37oC. After co-incubation, E. coli colonies developed on Levine media were counted towards the control. E. coli colonies decreased after 48 and 72h of co-incubation with L. plantarum. Also, was noticed that L. plantarum (108 CFU/mL) completely inhibits E. coli (104 CFU/mL) after 72h of co-incubation. This work proves the inhibitory potential of probiotic strains (L. plantarum ATCC 8014) against enteropathogenic bacteria (E. coli ATCC 25922).

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