1. NEISSERIA
Basmah almaarik
Lab # 6

2. NEISSERIA SPECIES Contains many species; 2 are human pathogens
Neisseria gonorrhoea (gonococcus)
Neisseria meningitidis (meningococcus)
Other species are normal flora that inhabit the upper respiratory tract of humans.

3. Process specimen as soon as possible
Neisseria gonorrhoea  Urethral discharge
 Genital swabs (servical or rectal)
Neisseria meningitidis  CSF
Fastidious organism very sensitive to environmental factors
Process specimen as soon as possible

4. GRAM STAIN Direct Gram Stain: Neisseria are intracellular organisms
GNDC inside and outside pus cells (PMN) (kidney or bean shaped)

5. CULTURE Inoculate into Chocolate Agar
Basal media  at 40C (no solidification) add sheep blood
Keep in water bath at 70C to hemolyse blood
This releases and denatures many RBCs and serum proteins,
making them more readily digestible to the bacteria
Colonial morphology: Grayish white, convex (raised), colony type varies, even in one strain, due to
antigenic variation
morphology:Grayish white, convex (raised), colony type varies, even in one strain, due to
anColony tigenic variation

6. Vancomycin, Colistin, Nystatin, Trimethoprim.
CULTURE
Thayer Martin (selective medium)
Chocolate agar contain antibiotics to inhibit other bacteria and fungi
VCNT
Vancomycin, Colistin, Nystatin, Trimethoprim.

7. Non-pathogenic Neisseria grow at room temperature
INCUBATION
Temp ˚C ONLY
Need 5-10% CO2 to grow:
CO2 envelop in anaerobic jar or
Candle jar
For 48 hours
Non-pathogenic Neisseria grow at room temperature

8. IDENTIFICATION Gram stain  GNDC Oxidase  +ve
Carbohydrate utilization test

9. IDENTIFICATION
Gram stain from plate  GNDC (Kidney shaped)

10. IDENTIFICATION All Neisseria species are Oxidase +ve
Preparation of Oxidase Test:
Add filter paper on a clean slide
Add 2-3 drops of oxidase reagent on filter paper
Use cover slip to transfer colony onto filter paper
+ ve  Purple color

11. Neisseria meningitides Neisseria gonorrhoeae
IDENTIFICATION
HISS medium.
Broth (CTA: Cystine trypticase agar)+ serum + sugar + indicator (phenol red)
Neisseria meningitides
Neisseria gonorrhoeae

12. YOUR WORK FOR TODAY Prepare gram stain from culture. Do oxidase test.
Subculture organism to chocolate agar.
Incubate in the CO2 jar.
Read after 48 hours.
See sugar utilization tube (demonstration)
See intracellular GNDC slide (demonstration slide)

13. http://student.ccbcmd.edu/~gkaiser/goshp. htm
New diagnosis techniques:
Immunoassays
DNA hybridization
PCR
Doc Kaiser’s microbiology website
htm

14. Numbers in Saudi Population
13 cases of meningococcal meningitis in 2007

15. Morexilla Gram stain GNDC culture 35-37c
Gorw on BA, CA and nutrient A.
Biochemical test
Oxidase +ve
No Acid production in Hiss medium test.

16. THANK YOU