Metabolite profiling of Clostridium difficile ribotypes using small molecular weight volatile organic compounds

Kuppusami, Sharmilah; Clokie, Martha R. J.; Panayi, Tolis; Ellis, Andrew M.; Monks, Paul S.

doi:10.1007/s11306-014-0692-4

Cited by 12 publications

(9 citation statements)

References 42 publications

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“…However, although all samples have been processes in the same way, hot spots of sulphur were seen only in the −10 °C culture. Sulphur may be of biological origin and be contained in hydrogen sulphide, carbon disulphide and other sulphur compounds that are known metabolic products of clostridia (Kuppusami et al ., ).…”

Section: Resultsmentioning

confidence: 97%

Epifluorescence, SEM, TEM and nanoSIMS image analysis of the cold phenotype ofClostridium psychrophilumat subzero temperatures

Perfumo

Elsaesser

Littmann

et al. 2014

FEMS Microbiol Ecol

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We have applied an image-based approach combining epifluorescence microscopy, electron microscopy and nanoscale secondary ion mass spectrometry (nanoSIMS) with stable isotope probing to examine directly the characteristic cellular features involved in the expression of the cold phenotype in the Antarctic bacterium Clostridium psychrophilum exposed to a temperature range from +5 to -15 °C under anoxic conditions. We observed dramatic morphological changes depending on temperature. At temperatures below -10 °C, cell division was inhibited and consequently filamentous growth predominated. Bacterial cells appeared surrounded by a remarkably thick cell wall and a capsule formed of long exopolysaccharide fibres. Moreover, bacteria were entirely embedded within a dense extracellular matrix, suggesting a role both in cryo-protection and in the cycling of nutrients and genetic material. Strings of extracellular DNA, transient cell membrane permeability and release of membrane vesicles were observed that suggest that evolution via transfer of genetic material may be especially active under frozen conditions. While at -5 °C, the bacterial population was metabolically healthy, at temperatures below -10 °C, most cells showed no sign of active metabolism or the metabolic flux was extremely slowed down; instead of being consumed, carbon was accumulated and stored in intracellular granules as in preparation for a long-term survival.

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Section: Resultsmentioning

confidence: 97%

Epifluorescence, SEM, TEM and nanoSIMS image analysis of the cold phenotype ofClostridium psychrophilumat subzero temperatures

Perfumo

Elsaesser

Littmann

et al. 2014

FEMS Microbiol Ecol

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“…difficile , since it continues to grow at concentrations which inhibit other bacteria [43]. There is evidence that the ability to produce and tolerate 4-methyphenol is even more pronounced in hypervirulent strains [44] and it might be possible distinguish these from other strains by their VOC profile [45].…”

Section: Discussionmentioning

confidence: 99%

Faecal volatile biomarkers of Clostridium difficile infection

et al. 2019

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Care of patients with potential CDI can involve isolation and use of antibiotics, often before a definitive diagnosis is available, impacting healthcare resource and contributing to antibiotic resistance. There is anecdotal evidence that the faeces of CDI patients have a distinctive odour, while it is well-established that changes in the gut microbiota are associated with changes in the volatile organic compounds (VOC) produced. A total of twenty-four candidate volatile biomarkers were identified from a review of the literature including in vitro , animal and human studies. Using thermal desorption-gas chromatography-time-of flight mass spectrometry (TD-GC-ToFMS), VOC emission rates were determined on stored frozen stool samples from 53 CDI-positive and 53 CDI-negative patients with unexplained diarrhoea which had previously been diagnosed using enzymatic and nucleic acid amplification tests. Sample preparation was limited to placement of a subsample in an appropriate container. Compounds exhibiting a statistically significant difference (p < 0.05) in emission rate between the CDI-positive and–negative groups and a corresponding area under the receiver-operator characteristic curve (ROC) >0.7 were considered potentially indicative of CDI. Seven compounds were so identified: propan-1-ol (ROC 0.75), 3-methylbutanal (ROC 0.84), ethyl propionate (ROC 0.81), hexanoic acid (ROC 0.73), 4-methylphenol (ROC 0.81), dodecane (ROC 0.80) and indole (ROC 0.85). A number of potential volatile biomarkers of CDI can be sampled rapidly and with little prior preparation from faecal samples of patients with diarrhoea. Of these 4-methylphenol ( p -cresol) is of particular interest as it has been anecdotally linked to CDI and is closely related to the biology and virulence of Clostridium difficile . This approach shows promise for the rapid, point-of-care diagnosis of CDI with good sensitivity and specificity.

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“…The production of p -cresol is a unique feature of C. difficile and closely related organisms, and has been exploited for diagnostic purposes in the past ( Sivsammye and Sims, 1990 ; Kuppusami et al, 2015 ). The frequent loss of p -cresol production observed here, however, questions the value of this trait for microbiological diagnostics.…”

Section: Discussionmentioning

confidence: 99%

Convergent Loss of ABC Transporter Genes From Clostridioides difficile Genomes Is Associated With Impaired Tyrosine Uptake and p-Cresol Production

et al. 2018

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We report the frequent, convergent loss of two genes encoding the substrate-binding protein and the ATP-binding protein of an ATP-binding cassette (ABC) transporter from the genomes of unrelated Clostridioides difficile strains. This specific genomic deletion was strongly associated with the reduced uptake of tyrosine and phenylalanine and production of derived Stickland fermentation products, including p-cresol, suggesting that the affected ABC transporter had been responsible for the import of aromatic amino acids. In contrast, the transporter gene loss did not measurably affect bacterial growth or production of enterotoxins. Phylogenomic analysis of publically available genome sequences indicated that this transporter gene deletion had occurred multiple times in diverse clonal lineages of C. difficile, with a particularly high prevalence in ribotype 027 isolates, where 48 of 195 genomes (25%) were affected. The transporter gene deletion likely was facilitated by the repetitive structure of its genomic location. While at least some of the observed transporter gene deletions are likely to have occurred during the natural life cycle of C. difficile, we also provide evidence for the emergence of this mutation during long-term laboratory cultivation of reference strain R20291.

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Metabolite profiling of Clostridium difficile ribotypes using small molecular weight volatile organic compounds

Cited by 12 publications

References 42 publications

Epifluorescence, SEM, TEM and nanoSIMS image analysis of the cold phenotype ofClostridium psychrophilumat subzero temperatures

Epifluorescence, SEM, TEM and nanoSIMS image analysis of the cold phenotype ofClostridium psychrophilumat subzero temperatures

Faecal volatile biomarkers of Clostridium difficile infection

Convergent Loss of ABC Transporter Genes From Clostridioides difficile Genomes Is Associated With Impaired Tyrosine Uptake and p-Cresol Production

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