Growth of Campylobacter pylori in liquid media

Abstract

Until recently, broth cultivation techniques for Campylobacter pylori were unavailable. We developed a method to cultivate bacterial cells within 24 h in liquid media. Cultivation in broth depended on the adequate dispersion of appropriate gases. A static broth at 37°C in a GasPak jar (BBL Microbiology Systems, Cockeysville, Md.) with a CampyPak (BBL) envelope did not support growth after 5 days of incubation. A broth placed in a flask on a Gyrotory water bath shaker (150 rpm; New Brunswick Scientific Co., Inc., Edison, N.J.) fitted with a gassing hood connected to a gas mixture of 10% C02, 5% 02, and 85% N2 supported good growth. An initial inoculum of 105, 103 to 104, or 102 CFU/ml resulted in >108 CFU/ml after incubation for 24, 48, or 72 h, respectively. Under these conditions, the bacteria grew as motile, spiral bacilli rather than the oval and coccal bacilli occasionally reported. Several bases supported good growth when supplemented with serum. For the determination of basal growth conditions, brucella broth base was used. Fetal calf serum (1%) provided maximum growth. Vitox was not necessary for growth and did not augment growth. C. pylori grew over a wide optimal pH range of 5.5 to 8.5.

Acid-responsive activity of the Helicobacter pylori metalloregulator NikR

“…H. pylori is an unusual component of the human gut microbiome, because it colonizes the acidic environment of the stomach (6,7). As a pathogen with a specific ecological niche, H. pylori has a compact genome and relatively few regulatory proteins (49).…”

“…H. pylori is unique because it primarily inhabits the human stomach (4), where it must survive a fluctuating pH that drops as low as 1.8 during digestion (5). However, H. pylori is a neutralophile (6,7), so to thrive in its ecological niche the bacteria must have robust mechanisms of acid adaptation (8). The focal component of pH homeostasis in H. pylori is urease, a dodecameric nickel enzyme that hydrolyzes host urea into ammonia to neutralize the internal pH as well as the local microenvironment (8).…”

A novel compound from celery seed with a bactericidal effect against <I>Helicobacter pylori</I>

“…[26] H. pylori was cultured at 37 C under microaerobic conditions (10% CO 2 , 3% O 2 and 87% N 2 ) on chocolate agar plates or in liquid Brucella broth (BBL, Franklin Lakes, NJ, USA) supplemented with foetal bovine serum (5% v/v; Invitrogen, Paisley, UK) immediately before use. [27] Antimicrobial activity test Microplate antimicrobial activity assays were performed using 24-well plates. Each assay contained 0.1 ml sample together with 1.0 ml early log phase H. pylori culture in Brucella broth plus foetal bovine serum (prepared as above).…”

Clinical Relevance of Culture: Why, How, and When

“…Growth in broth occurs with any of the basal media that is used also to prepare agars, such as Columbia, brain heart infusion, and Muller Hinton bases, as long as serum is added and the culture is shaken rather than remaining static. This, presumably, is due to the better gas transfer into the bulk medium [23]. In Brucella broth, bisulfite—present in the basal medium—is inhibitory for growth of H. pylori , and improved growth can be obtained if it is omitted [24].…”