In vitro inhibition of Clostridium difficile by commercial probiotics: A microcalorimetric study

Fredua‐Agyeman, Mansa; Stapleton, Paul; Basit, Abdul W.; Beezer, Anthony E.; Gaisford, Simon

doi:10.1016/j.ijpharm.2016.12.005

Cited by 30 publications

(19 citation statements)

References 42 publications

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“…In contrast, C. cellulolyticum reached highest sporulation efficiency at the lowest tested pH of 6.4 (54). In accordance with previous studies, no growth of C. difficile was observed below a pH of 5.5 (55–58). Comparative genomic analysis of C. difficile versus C. sordellii revealed the absence of several acid adaptation and broad range pH survival mechanism in C. difficile , such as ureases, glutamate decarboxylase, arginine deaminase or potassium transport proteins (59), which may explain the inability of C. difficile to grow under more acidic conditions.…”

Section: Discussionsupporting

confidence: 93%

The impact of pH onClostridioides difficilesporulation and physiology

Wetzel

McBride

2019

Preprint

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23 24 25 2 ABSTRACT 26Clostridioides difficile is a pathogenic bacterium that infects the human colon to cause 27 diarrheal disease. Growth of the bacterium is known to be dependent on certain bile 28 acids, oxygen levels and nutrient availability in the intestine, but how the environmental 29 pH can influence C. difficile is mostly unknown. Previous studies indicated that C. difficile 30 modulates the intestinal pH, and prospective cohort studies have found a strong 31 association between a more alkaline fecal pH and C. difficile infection. Based on these 32 data we hypothesized that C. difficile physiology can be affected by various pH 33 conditions. In this study, we investigated the impact of a range of pH conditions on 34 C. difficile to assess potential effects on growth, sporulation, motility and toxin production 35 in the strains 630∆erm and R20291. We observed pH-dependent differences in 36 sporulation rate, spore morphology and viability. Sporulation frequency was lowest under 37 acidic conditions, and differences in cell morphology were apparent at low pH. In alkaline 38 environments, C. difficile sporulation was greater for strain 630∆erm, whereas R20291 39 produced relatively high levels of spores in a broad range of pH conditions. Rapid 40 changes in pH during exponential growth impacted sporulation similarly among the 41 strains. Furthermore, we observed an increase in C. difficile motility with increases in pH, 42and strain-dependent differences in toxin formation under acidic conditions. The data 43 demonstrate that pH is an important parameter that affects C. difficile physiology and 44 may reveal relevant insights into the growth and dissemination of this pathogen. 45 46 47 IMPORTANCE 48 Clostridioides difficile is an anaerobic bacterium that causes gastrointestinal disease. C.49 difficile forms dormant spores, which can survive harsh environmental conditions, 50 allowing their spread to new hosts. In this study, we determine how intestinally relevant 51 3 pH conditions impact C. difficile physiology in the two divergent strains, 630∆erm and 52 R20291. Our data demonstrate that low pH conditions reduce C. difficile growth, 53 sporulation, and motility. However, toxin formation and spore morphology are 54 differentially impacted in the strains at low pH. In addition, we observed that alkaline 55 environments reduced C. difficile growth, but increased cell motility. When pH was 56 adjusted rapidly during growth, we observed similar impacts on both strains. This study 57 provides new insights into the phenotypic diversity of C. difficile grown under the diverse 58 pH conditions present in the intestinal tract, and demonstrates similarities and 59 differences in the pH responses of different C. difficile isolates. 60 61 62 INTRODUCTION 63 Clostridioides difficile is an emerging gastrointestinal pathogen, which often infects 64 patients who have recently received antibiotics. Upon ingestion, the dormant spores 65 survive the acidic pH of the stomach and enter the small intestine, where primary bile ...

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Section: Discussionsupporting

confidence: 93%

The impact of pH onClostridioides difficilesporulation and physiology

Wetzel

McBride

2019

Preprint

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“…), growth inhibition via the production of short‐chain fatty acids and other bioactive metabolites (Fredua‐Agyeman et al . ; Spinler et al . ; Najarian et al .…”

Section: Resultsmentioning

confidence: 99%

“…The emergence of highly virulent and antibiotic-resistant strains and the recent reports that C. difficile is a potential food-pathogen further increased the incidence and severity of CDIs. Lactic acid bacteria, especially Lactobacillus, which is also a member of the normal gut microbiota, have been reported to interfere with C. difficile toxin production via suppression of virulence factors and quorum-sensing system (Yun et al 2014;Najarian et al 2019), growth inhibition via the production of short-chain fatty acids and other bioactive metabolites (Fredua-Agyeman et al 2017;Spinler et al 2017;Najarian et al 2019) and prevention of C. difficile colonization via competitive exclusion (Dicks et al 2015;Najarian et al 2019). Since Lactobacillus is one of the most important gut microbiota; hence, acid and bile acid tolerance are important criteria for surviving the gastrointestinal tract.…”

Section: Resultsmentioning

confidence: 99%

Suppressive effect ofLactobacillus fermentumLim2 onClostridioides difficile027 toxin production

Yong

Lim

Kim

et al. 2019

Lett Appl Microbiol

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Clostridioides difficile is a spore‐forming, Gram‐positive, anaerobic pathogen that caused gastrointestinal illness. During dysbiosis, overgrowth of C. difficile resulting in higher levels of toxin production. Since Lactobacillus has been commonly used to alleviate gastrointestinal discomfort, this study aimed to investigate the effects of Lactobacillus isolated from kimchi on the quorum‐sensing and virulence factors of C. difficile 027. Among the isolated Lactobacillus strains, the acid and bile tolerant L. fermentum Lim2 was only able to reduce C. difficile 027 growth by one log10 CFU per ml. In keeping with this finding, C. difficile 027 growth was unaffected by either untreated or heat‐inactivated cell extracts from L. fermentum Lim2. Both untreated and heat‐inactivated cell extracts did, however, significantly reduce the autoinducer‐2 (AI‐2) activity of C. difficile 027, with the most prominent suppression effect (654‐fold) being found from 100 mg ml−1 of heat‐inactivated cell extract. A gene expression analysis indicated that in the presence of 100 mg ml−1 heat‐inactivated cell extract, the quorum‐sensing (luxS) and the virulence factors (tcdA, tcdB and tcdE) were significantly suppressed, whereas the negative regulator gene (tcdC) was significantly up‐regulated. Taken together, the significant anti‐pathogenic effect from L. fermentum Lim2 could potentially be used to treat C. difficile‐infections. Significance and Impact of the Study Clostridioides difficile is a Gram‐positive pathogenic bacteria that caused gastrointestinal illness via toxic production. The emergence of highly virulence and foodborne C. difficile strains has further increased the incident and severity of C. difficile‐infections (CDIs). Numerous studies have reported the immunomodulatory activity of Lactobacillus, a member of healthy gut microbiota, to maintain gastrointestinal health. Here, we successfully isolated L. fermentum Lim2 from kimchi, and identified a promising anti‐pathogenic effect against C. difficile 027, from the heat‐inactivated L. fermentum cell extract via suppression on the C. difficile 027 quorum‐sensing system and toxin production, which could potentially be used to treat and prevent CDIs.

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“…In addition to potentially ameliorating the clinical symptoms of gut disorders, a further potential application of probiotics is to counter infections caused by pathogenic species and immunomodulation (Fredua-Agyeman et al, 2017, Moens et al, 2019. Intestinal Infectious diseases caused by pathogenic organisms are major causes of mortality in both developed and developing countries (Nomoto, 2005).…”

Section: Introductionmentioning

confidence: 99%

Use of a water-based probiotic to treat common gut pathogens

Dodoo

Stapleton

Basit

et al. 2019

International Journal of Pharmaceutics

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This work reports the anti-pathogenic effect of a commercially available water-based probiotic suspension, Symprove, against three commonly encountered infectious organisms; Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA) and Shigella sonnei. An isothermal calorimetric assay was used to the monitor growth of the species individually and in binary combinations, while colony plate counting was used to enumerate viable cell numbers. It was observed that all pathogenic species were faster growing than the probiotic bacteria in Symprove after inoculation into growth medium yet in all instances bacterial enumeration at the end of the experiments revealed a significant reduction in the pathogen population compared with the controls. A control population between 10 8-10 9 CFU/ml was obtained for E. coli and S. sonnei whilst approximately 10 6 CFU/ml was obtained for MRSA. Upon co-incubation for 48 hours, no viable counts were obtained for E. coli; a 4-log reduction was obtained for S. sonnei whilst MRSA numbers were down to less than 10 cells/ml. The results show that Symprove has antipathogenic activity against E. coli, S. sonnei and MRSA.

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In vitro inhibition of Clostridium difficile by commercial probiotics: A microcalorimetric study

Cited by 30 publications

References 42 publications

The impact of pH onClostridioides difficilesporulation and physiology

The impact of pH onClostridioides difficilesporulation and physiology

Suppressive effect ofLactobacillus fermentumLim2 onClostridioides difficile027 toxin production

Use of a water-based probiotic to treat common gut pathogens

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