Gayam, Glydenne Glaire P.

BS Medical Technology 3A

Bacte MWF 5:30-8:30

August 17, 2015

Direct Gram Stain and Selection of Media for Bacterial Cultures

Specimen

Gram Stain

BAP

CHOC

MAC or EMB

Aqueous/Vitreous
Blood
Body Fluids

Amniotic
Bile
Bone Marrow
Pericardial
Peritoneal
Pleural
Synovial
Catheter Tips
CSF

X
X
X
X
X
X
X

ANA

THIO

TM

Other

X
Blood culture bottles
Blood culture bottles if
sufficient volume of fluid

X
X
X
X
X
X
X
X
X

X
X
X

X
X
X

X
X
X

X
X
X

X
X
X

X
X
X

X
X
X
X
X

X
X

X
X

X
X
X
X
X
X
X

Cytocentrifuge
recommended for Gram
Stain

Ear

Inner
Outer

Eye
Gastrointestinal
Duodenal Aspirate
Feces
Gastric Aspirate
Genital
IUD
Vaginal/Cervix
Urethra

X
X
X
X

X
X
CAN
HE or XLD, CAMPY, SMAC

X
X
X

X
X

X
X

Specimen
Genital Screens
Neisseria gonorrhoeae
Group B beta streptococci
Lesion/Wound/Abscess

Respiratory Tract: Lower

Bronchial (Brush/Wash/Lavage)
Sputum
Respiratory Tract: Upper
Nasal/Nasopharynx
Sinus Aspirate
Throat
Tissue
Urine
Catheter/Void
Suprapubic Aspirate

Gram Stain

BAP

CHOC

MAC or EMB

ANA

THIO

Other

X
X

X
X

X
X

X
X

X
X

X
X

X
X
X
X

X
X

TM

X
X

Lim broth* or carrot broth

CNA if Gram suggests
mixed Gram positive and
negative

*Lim broth, selective Todd Hewitt broth with CNA.

ANA, anaerobic culture media (anaerobic blood agar, Bacteroides bile esculin agar, kanamycin-vancomycin laked blood agar, anaerobic CNA or
PEA (phenylethyl alcohol agar); BAP, Sheep blood agar; CAMPY, Campylobacter-selective blood agar; CHOC, chocolate agar; CNA, Columbia
colistin-nalidixic acid agar; CSF, cerebrospinal fluid; EMB, eosin-methylene blue agar; HE, Hektoen-enteric agar; IUD, intrauterine device; MAC,
MacConkey agar; SMAC, sorbitol-MacConkey agar; THIO, thioglycollate broth; TM, Thayer-Martin or other Neisseria-selective agar; XLD, xyloselysine-deoxycholate agar.

Mahon, C.R. et. al (2015). Textbook of Diagnostic Microbiology (4th edition). Saunders Elsevier.

Gayam, Glydenne Glaire P.

BS Medical Technology 3A

Bacte MWF 5:30-8:30

August 17, 2015

Guidelines for Media Selection for Various Specimens

MEDIA FOR RECOVERY OF AEROBIC AND FACULTATIVELY ANAEROBIC BACTERIA

Specimen

BAP

MAC or
EMB

CBA

Broth

Peritoneal
Pleural; Pericardial
Synovial
Wounds
Aspirate
Swab
Tissue #
Respiratory Tract
Sputum
Throat
Bronchoalveolar
lavage
Brush; Washings
Nasal
Genitourinary
Vaginal/Rectal for

BAP

BBE

PEA

X
X

Consider the
use of BCB for
large volumes
of fluids

Body Cavities

Fluids
Cerebrospinal

Other

MEDIA FOR RECOVERY OF ANAEROBIC BACTERIA

X
X
X

X
X
X

X
X
X

X
X
X

X
X
X

X
X

Lim Broth

X (for shunt
specimens)
X
X

BCB
BCB
BCB

CYE

Selective or

group B
Streptococcus (GBS)
Other
Cervix
Urethra/Penis
Urine
Mild Void

chromogeni
c
X

Suprapubic Aspirate
Feces

Eye
Ear; Internal Aspirate
Vascular catheters

X
X
X

Screen;
chromagar
X

X
X

GBS media
GC media**
GC media
GC media

EB

HE or XLD;
Campy
X
X

BAP, Blood agar plate; BCB, blood culture bottles; BBE, Bacteroides bile esculin agar; Campy, Campylobacter-selective medium; CBA, chocolate
blood agar; CYE, charcoal yeast extract, for Legionella or Nocardia requests; EB, enrichment broth, such as GN or Selenite broth, same for
rectal swabs, minus the Campylobacter-selective culture; EMB, eosin methylene blue; GBS, group B streptococcus; GC, gonococcus; HE,
Hektoen enteric agar; Lim broth, enrichment broth for group B streptococcus; MAC, MacConkeys agar; PEA, phenylethyl alcohol; XLD,
xylose-lysine-deoxycholate agar.
*Specific guidelines for individual organisms will be included where they are described in text.

Supplemented thioglycollate in the usual broth; however, for aerobes, a brain-heart infusion may be adequate.

Consider a CDC BAP or a Brucella blood agar, or another enriched BAP for anaerobic recovery; a laked blood agar plate with antibiotics may
also be appropriate.

Not recommended for anaerobic cultures.

#
If specific organisms, or situations, other media may be added.

If a protected bronchoscope is used for collection of the specimen.

**
Thayer-Martin or Martin-Lewis or other media enriched for recovery of N. gonorrhoeae.

If Propionibacterium acnes is suspected in cases of endophthalmitis, a thioglycollate broth and/or anaerobic BAP may be used.

McPherson, R.A.;Pincus, M.R. (2012). Henrys Clinical Diagnosis and Management by Laboratory Methods (22 nd edition). Elsevier Saunders.