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1. Salahaddin University-Erbil College of Sciences General Biology Department,4 th stage (2025-2026 ) Medical Bactryology (practice) Date of Presentation 1/10/2025 Helicobacter pylori Prepared by: Rasty Abdullah Muhammad   Supervisor : Assis. Pro. Dr. Daristan Ghareb M. Ala Ahmad

2. Table of content Introduction Morphology Sample Collection Characteristic of Culture Media Growth Characteristic Biochemical test Identification Reference

3. Introduction Professor Marshall, and Dr Warren, are cited for their 1982 discovery of “the bacterium Helicobacter pylori and its role in gastritis and peptic ulcer disease.” What the brevity of that citation conceals is the extent to which their work opened the way to a simple cure for a common problem. It also turned an often performed surgical procedure into a relative rarity and stimulated the search for other microbes that may be the cause of chronic conditions.

4. Morphology in vivo and under optimum in vitro conditions is an S-shaped bacterium with 1 to 3 turns, 0.5 ×5 μm in length, with a tuft of 5 to 7 polar sheathed flagella . This morphology has been correlated with maximum in vitro motility . The majority of helicobacters possess this basic morphology of an S shape with polar, sheathed flagella, though variations in size and the number of spirals/turns are seen.

5. Morphology Thin sections of H. pylori reveal the typical cell wall detail of a gram-negative bacterium that consists of outer and inner, or plasma, membranes separated by the periplasm of approximately 30 nm thickness. The dense cytoplasm contains nucleoid material and ribosomes. Analysis of the peptidoglycan from H. pylori revealed it had a unique muropeptide composition, being less complex structurally than that seen in other gram-negative bacteria . Ultrathin section of  H. pylori  colonization of gastric mucus. Bar, 1 μm .

6. Characteristic of Culture Media

7. Growth Characteristic In general, primary cultures of H. pylori have less oxygen tolerance than most Campylobacter species, with a growth maximum at 3 to 7% of O2. H. pylori is usually grown in jars with gas-generation kits or a standard microaerobic atmosphere, in CO2 incubators or anaerobic chambers with a microaerobic atmosphere. Most studies with standardized atmospheres for culture of H. pylori have used 2 to 5% O2, 5 to 10% (optimal closer to 10%) CO2, and 0 to 10% H2 .

8. Biochemical Culture of H. pylori. (a) Growth of H. pylori from biopsy sample of chronic gastritis patient on specially formulated HP-agar medium. A red zone is formed around the bacterial colony due to a change in pH of the medium from acidic to alkaline by the hydrolysis of urea by urease enzyme. The plate was incubated for 65–70 hours at 371C and 5% CO 2. (b) Positive colonies of H. pylori from drinking water samples were applied on HP-agar medium to make replica plates. The colour of the medium is changed from yellow to red. The plate was incubated for 96 hours at 371C and 5% CO 2 . 

9. Identification Urea Breath Test (UBT) Stool Antigen Test Serology

10. Reference Hazell, S.L., Lee, A., Brady, L. and Hennessy, W. (2001) 'Morphology and Ultrastructure', in Mobley, H.L.T., Mendz , G.L. and Hazell, S.L. (eds.) Helicobacter pylori: Physiology and Genetics. Washington (DC): ASM Press. Goodwin, C.S. and Worsley , B.W. (2001) 'Basic Bacteriology and Culture', in Mobley, H.L.T., Mendz , G.L. and Hazell, S.L. (eds.) Helicobacter pylori: Physiology and Genetics. Washington (DC): ASM Press. Mobley, H.L.T. (2001) 'Urease', in Mobley, H.L.T., Mendz , G.L. and Hazell, S.L. (eds.) Helicobacter pylori: Physiology and Genetics. Washington (DC): ASM Press. 'Nobel prize is awarded to doctors who discovered H pylori' (2005) BMJ, 331(7520), p. 795. doi : 10.1136/bmj.331.7520.795-a. Safdar , S., Siddiqui, M.F., Ain, Q.U. and Bokhari , H. (2011) 'PCR assay targeting virulence genes of Helicobacter pylori isolated from drinking water and clinical samples in Lahore metropolitan, Pakistan', Journal of Water and Health, 9(1), pp. 208-216. doi : 10.2166/wh.2010.169. O’Rourke, J. and Bode, G. (2001) ‘Morphology and Ultrastructure’, in Mobley, H.L.T., Mendz , G.L. & Hazell, S.L. (eds.) Helicobacter pylori: Physiology and Genetics. Washington, DC: ASM Press.