agar and sodium-sulfite

Helicobacter pylori (H. pylori) is an obligate microaerobion and does not survive in low oxygen. Sodium sulfite (SS) reacts and consume oxygen in solutions. The present study aimed to investigate the effects of SS on H. pylori. The effects of SS on oxygen concentrations in solutions and on H. pylori in vivo and in vitro were examined, and the mechanisms involved were explored. The results showed that SS decreased the oxygen concentration in water and artificial gastric juice. In Columbia blood agar and special peptone broth, SS concentration-dependently inhibited the proliferation of H. pylori ATCC43504 and Sydney strain-1 in Columbia blood agar or special peptone broth, and dose-dependently decreased the number of H. pylori in Mongolian gerbils and Kunming mouse infection models. The H. pylori was relapsed in 2 weeks withdrawal and the recurrence in the SS group was lower than that in the positive triple drug group. These effects were superior to positive triple drugs. After SS treatments, the cell membrane and cytoplasm structure of H. pylori were disrupted. SS-induced oxygen-free environment initially blocked aerobic respiration, triggered oxidative stress, disturbed energy production. In conclusion, SS consumes oxygen and creates an oxygen-free environment in which H. pylori does not survive. The present study provides a new strategy and perspective for the clinical treatment of H. pylori infectious disease.

Topics: Agar; Animals; Disease Models, Animal; Gastric Mucosa; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Mice; Peptones

The present study was aimed at modifying the original formulation of Commercial Eugon agar (CEA) to develop a new H. pylori growth medium. Initial studies were carried out to determine the number of H. pylori colonies recovered on in-house H. pylori agar (IHPA), IHPA without L-cysteine and sodium sulfite (IHPA-NC), IHPA without L-cysteine (IHPA-C), IHPA without sodium sulfite (IHPA-N) and CEA as the control. Significant differences (P < 0.001) in the number of colonies recovered were observed between IHPA-N, IHPA-NC and IHPA-C. Incorporation of sodium sulfite decreased the number of colonies recovered, indicating that sodium sulfite was inhibitory to H. pylori growth. Removal of L-cysteine reduced the number of colonies recovered, suggesting that L-cysteine is necessary for the growth of H. pylori. In the subsequent study, incorporation of K(2)HPO(4) further increased the number of colonies recovered compared with IHPA-N (P < 0.001), and 0.25% (w/v) of K(2)HPO(4) yielded the highest numbers of colonies (P < or = 0.04). Finally, thirty other H. pylori clinical isolates were evaluated for their growth in the IHPAP-N, a new medium consisting of 1.5% (w/v) pepticase, 0.5% (w/v) peptone, 0.4% (w/v) sodium chloride, 0.03% (w/v) L-cysteine, 0.55% (w/v) dextrose, 0.25% (w/v) K(2)HPO(4) and 1.5% (w/v) agar. The number of colonies recovered in IHPAP-N was significantly (P < 0.005) higher than that of CEA. IHPAP-N with 0.25% K(2)HPO(4) and without sodium sulfite were adequate solid media for the growth of H. pylori.

Topics: Agar; Bacteriological Techniques; Colony Count, Microbial; Culture Media; Cysteine; Glucose; Helicobacter pylori; Peptones; Phosphates; Potassium Compounds; Sodium Chloride; Sulfites

A simple and rapid method for spore rec-assay by utilizing dry sheet medium culture (Compactdry TC, CTC) for determining numbers of bacteria, instead of the spore agar plate, was developed. One mL of spore suspension (2 x 10(6)/mL) of Bacillus subtilis strain M45 Rec- or H17 Rec+ was inoculated in the center of the CTC plate. In the case of metabolic activation, 1 mL of mixed solution (spore suspension of M45 or H17: 9,000 x g supernatant of rat-liver homogenate treated with Aroclor 1254 = 19:1) was used. The spore suspension spreads over the whole sheet in seconds and gels. A paper disk impregnated with 20-40 microL of the sample solution and 20 microL of the cofactor solution was placed on the surface of CTC plate. For the assay of samples that do not require metabolic activation, use of the cofactor solution can be omitted. After 48 hr incubation at 37 degrees C, 0.01% MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] aqueous solution (0.5 mL) was dropped uniformly on the plate. The plate was left for 5 min, and the diameter of the inhibition circle was measured with slide calipers. The samples for which the difference in inhibition zone between M45 and H17 was more than 2 mm were judged positive. Under these conditions, the DNA damaging activities of sodium sulfite, sodium benzoate and citric acid, used as food additives, were investigated by the proposed method. Sodium sulfite and sodium benzoate gave positive results and citric acid gave a negative result with or without metabolic activation, in agreement with the results obtained by the conventional method.

Topics: Agar; Bacillus subtilis; Bacteriological Techniques; Cell Count; Citric Acid; DNA, Bacterial; Food Additives; Sodium Benzoate; Spores, Bacterial; Sulfites