Survival of Helicobacter pylori in water and saline.

West, A. P.; Millar, Michael; Tompkins, David

doi:10.1136/jcp.43.7.609-b

Cited by 54 publications

(35 citation statements)

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“…Several factors may influence the spiral to coccoid conversion by H. pylori, such as acid pH stress, oxygen, temperature, nutritional starvation (West et al, 1990 ;Cellini et al, 1994 ;Donelli et al, 1998 ;Worku et al, 1999) and, thus, potentially culture media composition may influence the rate of spiral to coccoid conversion by Helicobacter spp. It has been speculated that the conversion to coccoid forms may be important for maintaining viability and survival of H. pylori outside the host (West et al, 1990), as well as in recrudescence of infection and treatment failure of patients with peptic ulcer disease (Benaissa et al, 1996). On the other hand, other microaerophilic bacteria produce non-culturable coccoid forms resembling those of helicobacters (Moran, 1997) and these have been shown to be a degenerate cell form which is undergoing cellular degradation (Moran & Upton, 1986) in response to oxygen toxicity and metabolites in culture media (Moran & Upton, 1987a, b).…”

Section: Introductionmentioning

confidence: 99%

“…This conversion generally occurs after 6-10 days of culture in stationary phase (Chan et al, 1994 ;Benaissa et al, 1996). Several factors may influence the spiral to coccoid conversion by H. pylori, such as acid pH stress, oxygen, temperature, nutritional starvation (West et al, 1990 ;Cellini et al, 1994 ;Donelli et al, 1998 ;Worku et al, 1999) and, thus, potentially culture media composition may influence the rate of spiral to coccoid conversion by Helicobacter spp. It has been speculated that the conversion to coccoid forms may be important for maintaining viability and survival of H. pylori outside the host (West et al, 1990), as well as in recrudescence of infection and treatment failure of patients with peptic ulcer disease (Benaissa et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Influence of activated charcoal, porcine gastric mucin and β-cyclodextrin on the morphology and growth of intestinal and gastric Helicobacter spp.

et al. 2002

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Bile-tolerant Helicobacter spp. are emerging human and animal pathogens. However, due to their fastidious nature, which requires nutrient-rich complex media to grow, infection with these bacteria may be underestimated. The accumulation of toxic metabolites in cultures may be one of the main obstacles for successful culture of these organisms. The present study examined various potential growth-enhancing substances for Helicobacter spp. and, furthermore, how they may affect spiral to coccoid conversion. Five Helicobacter spp. were cultured on agar and in broth media supplemented with activated charcoal, β-cyclodextrin, or porcine gastric mucin. Growth was determined by estimating the numbers of colony-forming units and colony diameter, as well as bacterial cell mass. Coccoid transformation was estimated every 24 h by both Gram and acridine-orange staining. Activated charcoal was superior in supporting growth and increased cell mass on agar and in broth media. β-Cyclodextrin delayed spiral to coccoid conversion by Helicobacter pylori and Helicobacter canis, whereas activated charcoal delayed the conversion to coccoid forms of Helicobacter hepaticus and Helicobacter bilis. The progression to coccoid forms by Helicobacter pullorum on agar media was not influenced by any growth supplement. The spiral to coccoid conversion was more rapid in broth media than on agar media. The growth enhancement observed is probably related to the capacity of activated charcoal to remove toxic compounds in culture media.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Influence of activated charcoal, porcine gastric mucin and β-cyclodextrin on the morphology and growth of intestinal and gastric Helicobacter spp.

et al. 2002

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“…They are particularly important in demonstrating the rapidity with which these forms appear, even at concentrations of amoxicillin that are in excess of 500-fold higher than the measured MIC. Coccoid forms, although metabolically quiescent, have been shown to retain important life functions (1,6,8,9) and have therefore aroused speculation that they may assume a significant role in therapy failure and the recrudescence of infection (2, 3, 7). H. pylori is susceptible to most antibiotics in vitro, including amoxicillin, but these agents per se have little effect in the clinic.…”

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Bactericidal and morphological effects of amoxicillin on Helicobacter pylori

Berry¹,

Jennings²,

Woodnutt³

1995

Antimicrob Agents Chemother

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The growth kinetics of Helicobacter pylori after it has been exposed to amoxicillin have been investigated in conjunction with studies of cell morphology. A potent bactericidal effect was observed at concentrations 10-fold higher than the MIC, but this was accompanied by an increase in the residual numbers of coccoid forms observed. In the presence of 10 g of amoxicillin per ml, these forms could be detected as rapidly as 6 h after exposure to the antibiotic. Although the clinical relevance of coccoid forms remains unknown, such forms should be considered when potential anti-Helicobacter agents are tested in vitro.Helicobacter pylori exists in several morphologically distinct forms, including nonculturable coccoid forms (3, 5). Coccoid formation has been attributed to environmental stress, including nutrient deprivation, extended incubation, accumulation of metabolic products, pH alteration, and exposure to antimicrobial agents (1,3,5). The last may be of major importance when therapy is considered, but the activity of agents in vitro is often determined only by susceptibility tests, e.g., agar dilution, in which the morphological status of the organism is unknown. Recent studies have demonstrated that amoxicillin at concentrations close to the MIC can result in the development of coccoid forms after 72 h of incubation (2). We describe observations on the time course of the bactericidal and morphological effects of a range of concentrations of amoxicillin on H. pylori.A broth cultivation technique similar to that described previously by Iwahi and coworkers (4) was used. Briefly, stock cultures of H. pylori NCTC 11637 were inoculated into 200 ml of Brucella broth (BBL) with 2.5% fetal calf serum (Gibco) in 500-ml sterile tissue culture flasks. After 16 h of microaerobic incubation (Gaspak jars and Campypaks; BBL) with gyration at 100 rpm, the culture (approximately 5 log 10 CFU/ml) was divided into 36-ml aliquots in 80-cm 2 tissue culture flasks, and 4-ml volumes of fresh medium or amoxicillin (SmithKline Beecham Pharmaceuticals, Worthing, England) prepared in broth to give concentrations of 0.01, 0.1, 1.0 and 10 g/ml were added. The MIC of amoxicillin for H. pylori NCTC 11637, as determined by agar dilution, was 0.015 g/ml. The cultures were reincubated as described previously, and at times from 0 to 36 h, 0.5-ml volumes were removed for the enumeration of viable bacteria, measurement of residual antibiotic concentration, and electron microscopy. The viability was determined by a modified Miles-Misra technique of preparing serial 10-fold dilutions in phosphate-buffered saline (PBS) and inoculating 20-l samples in triplicate onto Mueller-Hinton agar (Oxoid Ltd.) supplemented with 7% defibrinated horse blood. Colonies were counted after 3 days of incubation at 37ЊC under microaerophilic conditions and were expressed as CFU per milliliter. Samples, taken at random, were also plated onto nonselective media to determine purity, and organisms other than H. pylori were not detected. A large-plate agar diffusion techniq...

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“…The data indicated that transmission of H. mustelae is fecal-oral (4,11,12). H. pylori also has been cultured from feces and may survive in water in a nonculturable but viable coccoid form (10,30,31). The other proposed route of transmission is oral-oral (2,19,21,31).…”

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confidence: 99%

“…H. pylori also has been cultured from feces and may survive in water in a nonculturable but viable coccoid form (10,30,31). The other proposed route of transmission is oral-oral (2,19,21,31). H. pylori has also been cultured from saliva and dental plaques from humans, which argues for an oral-oral transmission (2,19).…”

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confidence: 99%

Identification of NovelHelicobacterspp. from a Beluga Whale

Harper

Feng

et al. 2002

Appl Environ Microbiol

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The gastric fluid and feces of three belugas from the Mystic Aquarium were assessed for the presence of Helicobacter spp. Gastric fluid and feces from the two clinically healthy belugas were negative for helicobacter, and endoscopy performed on these animals revealed no lesions. However, a helicobacter isolate and PCR product similar to helicobacter strains previously recovered from dolphins were identified, respectively, from the feces and gastric fluid of a beluga manifesting intermittent inappetence and lethargy. Esophageal and forestomach ulcers were noted on endoscopy. This is the first report of novel Helicobacter spp. being identified from whales.Belugas are a circumpolar species with an endangered relic population living in the Saint Lawrence River Estuary (17,18,23). Three belugas (animals 1, 2, and 3) cohoused at the Mystic Aquarium were assessed for helicobacter infection. Animal 1 manifested clinical signs, including intermittent inappetence and lethargy, and esophageal and forestomach ulcers were observed at endoscopic examination. Animals 2 and 3 showed no clinical signs, nor were esophageal and forestomach ulcers diagnosed endoscopically.Rectal swabs and gastric fluid were obtained from all three belugas for culture and PCR. A sterile swab (Becton Dickinson and Company, Sparks, Md.) was inserted through the anus to a depth sufficient to coat the swab with feces. The swab was then removed and placed in vials containing 20% glycerol in brucella broth. The gastric fluid was collected by endoscopy (Olympus 210-cm Videoscope) and aliquoted in individual one-dram vials. The endoscope and its channels were rinsed sequentially in dilute chlorhexidine, 70% alcohol, and water between each sampling. A Massachusetts Institute of Technology (MIT) accession number was assigned to each fecal and gastric fluid sample collected: samples from animal 1 were MIT 00-7128 and MIT 00-7129, those from animal 2 were MIT 00-7125 and MIT 00-7126, and those from animal 3 were MIT 00-7131 and MIT 00-7132. These sample numbers are used throughout this work.The fecal and gastric fluid samples used for microaerobic culture were placed in individual vials with 3 ml of 20% glycerol in brucella broth. The samples were placed on dry ice and then stored at Ϫ70°C prior to culture. The media used for culture were Trypticase soy agar with 5% sheep blood and TVP (trimethoprim, vancomycin, and polymyxin) and CVA (cefoperazone, vancomycin, and amphotericin B) antibioticimpregnated media (Remel Laboratories, Lenexa, Kans.). In addition, selective antibiotic medium (ABM) was prepared with blood agar base (Oxoid; Remel), 5% horse blood (Remel), amphotericin B (50 g/ml), vancomycin (100 g/ml), polymyxin B (3.3 g/ml), bacitracin (200 g/ml), and nalidixic acid (10.7 /ml) (all antibiotics from Sigma Chemical Co., St. Louis, Mo.). Approximately 0.5 g of feces was homogenized in 1 ml of brucella broth (Difco Laboratories, Detroit, Mich.) containing 5% fetal calf serum (Summit Technologies, Fort Collins, Colo.) in a glass tissue grinder. Approximate...

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Survival of Helicobacter pylori in water and saline.

Abstract: of "lumpectomy" for breast cancer. Hum Pathol 1986;17:330-2.

Cited by 54 publications

References 3 publications

Influence of activated charcoal, porcine gastric mucin and β-cyclodextrin on the morphology and growth of intestinal and gastric Helicobacter spp.

Influence of activated charcoal, porcine gastric mucin and β-cyclodextrin on the morphology and growth of intestinal and gastric Helicobacter spp.

Bactericidal and morphological effects of amoxicillin on Helicobacter pylori

Identification of NovelHelicobacterspp. from a Beluga Whale

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