Helicobacter Infections

Journals on Emerging Infectious Diseases

• CDC's journal, Emerging Infectious Diseases
• Helicobacter, The Journal (non-federal Web site)
• CDC's Public Health Image Library

Murine Helicobacter

Helicobacter organisms are of great current interest. There are approximately 20 Helicobacter species described in many species, including humans. A Web site is devoted to the human pathogen, H. pylori, which is also found in cats and monkeys, .

Dr. Barry Marshall's Helicobacter Web site (non-federal Web site)

H. heilmannii

H. muridarum from Dr. Lee

At NCI, in 1994, we found a new Helicobacter, H. hepaticus, in the mouse liver and large intestine. The bacteria can be seen with Steiner stain (figure below left). This new bacteria in bile canaliculi between hepatocytes may interfere with research studies.

How We Found The New Mouse Helicobacter

News from Japan about our new mouse liver Helicobacter.
 

The genome of Helicobacter hepaticus

Steiner stain showing H. hepaticus in bile canaliculi of mouse liver.

ABC immunohistochemistry showing H. hepaticus in bile canaliculi (between hepatocytes) of mouse liver using rabbit antibody to H. pylori.

Severe cholangitis and bile ductular hyperplasia in liver of mice with H. hepaticus infection.

Modified Steiner Technique for Detection of Helicobacter sp. in Tissue Sections

(from Garvey et al., J. Histotechnol. 8:15-17, 1985, modified by Dr. Miriam Anver and Cathi George, NCI-FCRDC, Frederick, MD)

• Fixation: 10% buffered neutral formalin
• Process: paraffin, frozen, or celloidin
• 1% Uranyl Nitrate: uranyl nitrate, 1.0 gm; distilled water, 100.0 ml
• 1% Silver Nitrate: silver nitrate crystals, 1.0 gm; distilled water, 100.0 ml
• 2.5% Gum Mastic: gum mastic, 2.5 gm; absolute alcohol, 100.0 ml
• 0.04% Silver Nitrate: silver nitrate crystals, 0.04 gm; distilled water, 100.0 ml
• 2% Hydroquinone: hydroquinone crystals, 1.0 gm; distilled water 50.0 ml
• Reducing solution (prepared just before use): 20.0 ml 2.5% gum mastic,  50 ml 2% hydroquinone, 10.0 ml 100% ethyl alcohol.  Combine the gum mastic solution and the hydroquinone solution. Add the alcohol and mix well. Filter through Whatmann #4 filter paper. Immediately before placing slides in solution, add 5 ml of 0.04% silver nitrate.
• Staining Procedure:
1. Decerate slides in xylene, 2 changes, 2 minutes each. Place in absolute alcohol, 2 changes, 2 minutes each; 95% alcohol, 2 changes, 2 minutes each; then rinse in distilled water.
2. Place in 1% uranyl nitrate, preheated to 60ºC., for 15 minutes.
3. Rinse thoroughly in distilled water.
4. Place in 1% silver nitrate for 2 hours at 60ºC.
5. Rinse in 2 changes of distilled water.
6. Dehydrate in 2 changes of 95% and 2 changes of 100% alcohol.
7. Place in 2.5% gum mastic for 5 minutes.
8. Allow sections to air dry for 1 minute or until white all over.
9. Rinse in 2 changes of distilled water.
10. Place sections in reducing solution that has been preheated to 43ºC (no more than 10 minutes). Check development on microscope after 10 minutes, then every 5 minutes until developed. Approximately 15-25 minutes.
11. Rinse in distilled water to stop reduction.
12. Dehydrate slides in 95% alcohol and absolute alcohol, 3 changes each. Clear in xylene, 3 changes.
13. Mount coverglass with appropriate medium (refractive index 1.48-1.56).
• Helicobacter sp., spirochetes, fungi, and most bacteria will appear dark brown to black.
• Background will be yellow to brown.

Ultrastructure (with 3D embossing) of mouse liver showing Helicobacter hepaticus in bile canaliculus of mouse liver.


In situ hybridization of liver showing bacteria 16S rRNA in liver but not in area of tumor


Marked rectal hyperplasia with invasion of submucosa.


Scanning EM photo of H. hepaticus within cecal crypts. (courtesy of Dr. John Sundberg, Jackson Labs)


Negative stain of H. bilus in culture. Note multiple flagellae and periplasmic fibrils.

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