AEROBIC GRAM NEGATIVE COCCI

Mahon & Manuselis, 2nd edition, chapter 14, pp. 401-423

General Characteristics

Habitat upper respiratory tract, alimentary and genitourinary tracts

Gram negative cocci occurring in pairs (diplococci) having a coffee bean or kidney bean
shape (GNDC)
Cytochrome oxidase positive, catalase positive, non-motile

Organisms
A. Neisseria species
1. Neisseria gonorrhoeae
a. Morphology and characteristics
i. Growth requirements
Fastidious (requiring an enriched media such as chocolate agar and 2-8% CO2 to
grow)
Temperature sensitive
Specimen must be inoculated to agar immediately
If long transportation time, send in transport media
ii.

Gram stain
From specimen: gram negative intracellular diplococci (GNID)
Isolated colony: GNDC

iii. Colony morphology

Chocolate agar
Thayer Martin agar - growth
o Selective for N. gonorrhoeae, N. meningitidis, and N. lactamica
o Agar is chocolate agar with antibiotics (vancomycin, colistin, and nystatin)
added
iv. Identification
Carbohydrate utilization
o Dextrose = positive
o Maltose = negative
o Lactose = negative
o Sucrose = negative
DNase = negative
Nitrate reduction = negative
Butyrate esterase (Catarrhalis disk) = negative
b. Pathogenesis
i. Port of entry
Primary infection usually occurs at columnar epithelium of urethra and
periurethral ducts, and glands of either sex
Cervical, conjunctival and rectal mucosa may also serve as portal of entry

CLS 418 Clinical Microbiology I

Student Laboratory
Aerobic Gram Negative Cocci

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ii.

Progression of infection
Three to five day incubation period
Mucosal --> submucosal -->inflammation -->abscesses and cysts --> lymphatic
vessels --> (sometimes blood vessels) bacteremia --> gonococcal arthritis
dermatitis --> rarely subacute bacterial endocarditis and meningitis

iii. Human to human transfer, primarily venereal spread

iv. Asymptomatic female reservoir
c.

Disease states
i. Gonorrhea
Male: acute urethritis, prostatitis and epididymitis if untreated
Female: may be asymptomatic or exhibit severe discharge
o Cervicitis --> pelvic inflammatory disease --> scarring of fallopian tubes -->
sterility, ectopic pregnancies
ii.

Disseminated gonococcal infection (DGI)

Spread from the genitourinary tract, rectum or pharynx to the blood stream

iii. Pharyngitis
Oral infection
iv. Gonococcal ophthalmia neonatorum
Conjunctiva infection in the newborn
v.

Vulvovaginitis
In young girls

d. Treatment
i. Empiric therapy options.
ii. Most patients treated with antibiotics for possible co-infections with Chlamydia.
iii. Eye treatments for infants with 1 hour of delivery
e. Antimicrobial testing
i. Standardized methods available but not routinely performed.
ii. Beta lactamase testing routinely done (ceftriaxone effective remains effective against
beta lactam producing isolates)
2. Neisseria meningitidis
a. Morphology and characteristics
i. Growth requirements
Fastidious (requiring an enriched media such as chocolate agar and 2-8% CO2 to
grow)
Temperature sensitive
Occasionally grows on sheep blood agar
ii.

Colony morphology
Chocolate agar
Thayer Martin agar - growth

CLS 418 Clinical Microbiology I

Student Laboratory
Aerobic Gram Negative Cocci

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iii. Identification
Carbohydrate utilization
o Dextrose = positive
o Maltose = positive
o Lactose = negative
o Sucrose = negative
DNase = negative
Nitrate reduction = negative
Butyrate esterase (Catarrhalis disk) = negative
b. Disease states
i. Meningitis, septicemia, septic arthritis, endocarditis, pneumonia, urethritis
ii.
c.

Asymptomatic nasopharyngeal carriers

Treatment and Antimicrobial testing

i. Standardized methods available but not routinely performed.
ii. Beta lactamase production is rare. Resistance by altered penicillin-binding proteins
is emerging.
iii. Drug of choice is penicillin.

d. Prevention
i. Single dose vaccine is available to polysaccharide capsular antigens A, C, Y and
W135.
ii. Chemoprophalyxis treatment given to close contacts of patient with meningococcal
meningitis.
Rifampin, ciprofloxin, or ceftriaxone
3. Neisseria lactamica
a. Morphology and characteristics
i. Colony morphology
Chocolate agar
Thayer Martin agar - growth
ii.

Identification
Carbohydrate utilization
o Dextrose = positive
o Maltose = positive
o Lactose = negative
o Sucrose = negative
DNase = negative
Nitrate reduction = negative
Butyrate esterase (Catarrhalis disk) = negative

b. Normal flora

B. Other Gram Negative Cocci

1. Moraxella (Branhamella) catarrhalis
a. Morphology and characteristics
i. Colony morphology
Sheep blood agar grows well
Chocolate agar grayish-white, granular colony
Thayer Martin agar variable growth
CLS 418 Clinical Microbiology I
Student Laboratory
Aerobic Gram Negative Cocci

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ii.

Identification
Carbohydrate utilization
o Dextrose = negative
o Maltose = negative
o Lactose = negative
o Sucrose = negative
DNase = positive
Nitrate reduction = positive
Butyrate esterase (Catarrhalis disk) = positive

b. Disease states
i. Acute bronchitis, pneumonia, meningitis, seen in immunocompromised patients
(elderly)
ii.
c.

In children otitis media, maxillary sinusitis

Antimicrobial Testing
i. Beta lactamase testing, most do not produce resistance through other mechanisms
so Beta lactamase testing is sufficient. Most are beta lactamase positive.
ii. Susceptibility testing not needed to guide therapy, many beta-lactam antibiotics
maintain activity.

Neisseriaceae
Gram negative cocci (diplococci)

Oxidase positive

Setup CTA sugars, Butyrate esterase disk, and check for growth on Thayer-Martin agar
Organism

Glucose

Maltose

Lactose

Butyrate

TM

Neisseria gonorrhoeae

Positive

Negative

Negative

Negative

Growth

Neisseria meningitidis

Positive

Positive

Negative

Negative

Growth

Neisseria lactamica

Positive

Positive

Positive

Negative

Growth

Moraxella catarrhalis

Negative

Negative

Negative

Positive

No growth

Note:

The only organisms that usually grow well on Thayer-Martin are N. gonorrhoeae, N.
meningitidis and N. lactamica.

Note:

Of the organisms that usually grow well on Thayer-Martin, only N. gonorrhoeae and
N. meningitidis are considered pathogenic.

Note:

There are many other Neisseria species. Due to their lack of clinical significance
they are reported as Neisseria species and/or considered normal flora.

Additional resources:
Koneman, Color Atlas and Textbook of Diagnostic Microbiology, 5th Edition, pages 491-524
Table 10-7, pages 516-517
Bailey & Scotts Diagnostic Microbiology, 11th edition, pages 505-508
Table 45-4, page 507
CLS 418 Clinical Microbiology I
Student Laboratory
Aerobic Gram Negative Cocci

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