Susceptibility of archaea to antimicrobial agents: applications to clinical microbiology

Khelaifia, Saber; Drancourt, Michel

doi:10.1111/j.1469-0691.2012.03913.x

Cited by 94 publications

(62 citation statements)

References 87 publications

(130 reference statements)

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“…These organisms are no longer considered prokaryotic organisms and share some similarities with the eukaryotic cells 7. They have some unique properties: live in strictly anaerobic conditions,8 difficult cultivation, requiring specific conditions and media,9 and cell wall lacking peptidoglycan making them therefore susceptible to certain antibiotics (i.e., chloramphenicol and bacitracin) and resistant to many common others like penicillins and aminoglycosides 10. The genetic diversity of methanogens in humans is limited11; only 3 phylotypes have been detected: Methaninobrevibacter smithii is the dominant methanogen in the intestine followed by Methanospaera stadmagnae 12 whereas Methannobrevibacter oralis which is implicated in periodontal disease is the main archaeon in the oral cavity 13.…”

Section: Methanogensmentioning

confidence: 99%

Methanogens, Methane and Gastrointestinal Motility

Triantafyllou

Chang

Pimentel

2014

J Neurogastroenterol Motil

208

152

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Anaerobic fermentation of the undigested polysaccharide fraction of carbohydrates produces hydrogen in the intestine which is the substrate for methane production by intestinal methanogens. Hydrogen and methane are excreted in the flatus and in breath giving the opportunity to indirectly measure their production using breath testing. Although methane is detected in 30%-50% of the healthy adult population worldwide, its production has been epidemiologically and clinically associated with constipation related diseases, like constipation predominant irritable bowel syndrome and chronic constipation. While a causative relation is not proven yet, there is strong evidence from animal studies that methane delays intestinal transit, possibly acting as a neuromuscular transmitter. This evidence is further supported by the universal finding that methane production (measured by breath test) is associated with delayed transit time in clinical studies. There is also preliminary evidence that antibiotic reduction of methanogens (as evidenced by reduced methane production) predicts the clinical response in terms of symptomatic improvement in patients with constipation predominant irritable bowel syndrome. However, we have not identified yet the mechanism of action of methane on intestinal motility, and since methane production does not account for all constipation associated cases, there is need for high quality clinical trials to examine methane as a biomarker for the diagnosis or as a biomarker that predicts antibiotic treatment response in patients with constipation related disorders.

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Section: Methanogensmentioning

confidence: 99%

Methanogens, Methane and Gastrointestinal Motility

Triantafyllou

Chang

Pimentel

2014

J Neurogastroenterol Motil

208

152

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“…Monitoring the M. oralis load by using real-time PCR could be useful for the diagnosis and staging of patients with periodontitis and treatment follow-up. It could complement clinical evaluation in detecting individuals at higher risk of developing severe periodontitis and be useful in evaluating the efficacy of antibiotic treatment, since M. oralis is highly resistant to antibiotics, except for metronidazole (19). The total bacterial DNA was tested by real-time PCR, and a quantification plasmid (Stdint) was introduced as an internal control to monitor the absence of PCR inhibition.…”

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confidence: 99%

Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

et al. 2013

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bA real-time PCR assay developed to quantify Methanobrevibacter oralis indicated that its inoculum significantly correlated with periodontitis severity (P ‫؍‬ 0.003), despite a nonsignificant difference in prevalence between controls (3/10) and patients (12/22) (P ‫؍‬ 0.2, Fisher test). The M. oralis load can be used as a biomarker for periodontitis. Periodontitis is an anaerobic infection possibly leading to loss of teeth (1-3). It likely results from infection with microbial complexes comprising methanogens (4, 5), mostly Methanobrevibacter oralis (6)(7)(8). In this study, we correlated the M. oralis load as measured by real-time PCR in subgingival plaque with the severity of periodontitis.All patients diagnosed with periodontitis from October 2011 to June 2012 were included. The control individuals with generally healthy gingiva were volunteers. All patients underwent interviews for medical and dental history, intraoral examination to determine bleeding on probing (BOP), probing depth (PD), and plaque index (PI), and radiographs (9); periodontal status was also scored as described previously (10) ( Table 1). All individuals signed an informed-consent form, and the ethics committee of the IFR 48, University of Aix-Marseille, approved the protocol.Subgingival plaque specimens (50 l) collected from 3-to 12-mm periodontal pockets in both patients and controls were suspended in 1 ml Tris-HCl (0.05 M, pH 7.5) buffer. After homogenization, a 250-l aliquot was shaken with 0.3 g of acid-washed beads (Յ106 mm; Sigma, Saint-Quentin-Fallavier, France) in a FastPrep-24 instrument (MP Biomedical Europe, Illkirch, France) at a speed of 6.5 m/s (full speed) for 90 s. The supernatant was incubated overnight at 56°C with 180 ml of lysis buffer and 25 l proteinase K (20 mg/ml) in the Qiagen EZ1 DNA tissue kit (Qiagen, Courtaboeuf, France). After a second cycle of mechanical lysis, the supernatant was incubated for 10 min at 100°C, and total DNA was then extracted using the same kit. Negative controls consisting of sterile DNA-free water were introduced in all the manipulations. The specificity of the M. oralis-cnp602P probe (6-carboxyfluorescein [FAM]-5=AGCAGTGCACCTGCTGATA TGGAAGG-3=) (Applied Biosystems, Courtaboeuf, France) and the primer pair M. oralis-cnp602F (5=-GCTGGTGTAATCGAAC CTAAACG-3=) and M. oralis-cnp602R (5=-CACCCATACCCGG ATCCATA-3=) (Eurogentec, Seraing, Belgium) was verified in silico using the BLAST program at NCBI (http://www.ncbi.nlm .nih.gov/BLAST) and further experimentally ensured by incorporating DNA extracted from Methanobrevibacter smithii ATCC 35061 T , Methanosphaera stadtmanae ATCC 43021 T , Methanomassiliicoccus luminyensis CSURP135 T , Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Treponema denticola ATCC 35405T . Real-time PCR assays were performed with a CFX96 Touch real-time PCR detection system (Bio-Rad, Marnes-laCoquette, France) using the Mastermix PCR kit (Eurogentec) with 5 pmol of each primer and probe and 5 l of about 2 g of DNA into a 20-l final volume. M. oralis DSMZ ...

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“…This contamination could be problematic because archaea significantly differ from bacteria so that the archaeacidal activity of biocides cannot be simply deduced from their bactericidal activity. Moreover, human-associated archaea have been found to be highly resistant to most commonly used antibiotics [14], [15]. In the perspective of broadening the spectrum of new active molecules, and more precisely against archaea organisms colonizing the human gut, squalamine and its derivatives appear to be among the few antimicrobial agents able to demonstrate an efficient anti-archaea activity [14], [15].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, human-associated archaea have been found to be highly resistant to most commonly used antibiotics [14], [15]. In the perspective of broadening the spectrum of new active molecules, and more precisely against archaea organisms colonizing the human gut, squalamine and its derivatives appear to be among the few antimicrobial agents able to demonstrate an efficient anti-archaea activity [14], [15]. Squalamine is a natural aminosteol compound which is extracted from the spiny dogfish shark live [16].…”

Section: Introductionmentioning

confidence: 99%

In-Vitro Archaeacidal Activity of Biocides against Human-Associated Archaea

2013

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BackgroundSeveral methanogenic archaea have been detected in the human intestinal microbiota. These intestinal archaea may contaminate medical devices such as colonoscopes. However, no biocide activity has been reported among these human-associated archaea.MethodologyThe minimal archaeacidal concentration (MAC) of peracetic acid, chlorhexidine, squalamine and twelve parent synthetic derivatives reported in this study was determined against five human-associated methanogenic archaea including Methanobrevibacter smithii, Methanobrevibacter oralis, Methanobrevibacter arboriphilicus, Methanosphaera stadtmanae, Methanomassiliicoccus luminyensis and two environmental methanogens Methanobacterium beijingense and Methanosaeta concilii by using a serial dilution technique in Hungates tubes.Principal FindingsMAC of squalamine derivative S1 was 0.05 mg/L against M. smithii strains, M. oralis, M. arboriphilicus, M. concilii and M. beijingense whereas MAC of squalamine and derivatives S2–S12 varied from 0.5 to 5 mg/L. For M. stadtmanae and M. luminyensis, MAC of derivative S1 was 0.1 mg/L and varied from 1 to ≥10 mg/L for squalamine and its parent derivatives S2–S12. Under the same experimental conditions, chlorhexidine and peracetic acid lead to a MAC of 0.2 and 1.5 mg/L, respectively against all tested archaea.Conclusions/SignificanceSqualamine derivative S1 exhibited a 10–200 higher archaeacidal activity than other tested squalamine derivatives, on the majority of human-associated archaea. As previously reported and due to their week corrosivity and their wide spectrum of antibacterial and antifungal properties, squalamine and more precisely derivative S1 appear as promising compounds to be further tested for the decontamination of medical devices contaminated by human-associated archaea.

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Susceptibility of archaea to antimicrobial agents: applications to clinical microbiology

Cited by 94 publications

References 87 publications

Methanogens, Methane and Gastrointestinal Motility

Methanogens, Methane and Gastrointestinal Motility

Real-Time PCR Quantification of Methanobrevibacter oralis in Periodontitis

In-Vitro Archaeacidal Activity of Biocides against Human-Associated Archaea

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