However, SB and HB are often not readily available to laboratories in resource-poor countries, either due to a lack of reliable sources of defibrinated blood
or due to financial constraints.
Almost 2 liters of defibrinated blood
was drained but no active bleeding source was found.
The isolates were cultured in Brucella agar medium (Merck, Germany) containing 10% sheep defibrinated blood
and 7% fetal bovine serum (FBS) (Gibco, UK, Manchester) at 37 [degrees]C for 72 hours under microaerophilic condition (10% CO2, 5% 02, 90% humid) in an CO2 incubator (Memmert, Germany) (9).
Aliquots of 0.1 ml of serial dilutions were plated in duplicate onto a) enriched trypticase soy agar (ETSA) supplemented with 4% defibrinated blood
to determine the predominant cultivable microbiota [Syed et al., 1980]; b) KVLB-2 agar (kanamycin 75[micro]g/ml-vancomycin 2[micro]g/ ml blood) for the isolation of pigmented and non-pigmented Prevotella spp, Porphyromonas spp and Bacteroides spp; c) TSBV (trypticase-soy-agar) supplemented with 5% serum 75 [micro]g/ml bacitracin and 5 [micro]g/ml vancomycin to quantify Aggregatibacter actinomycetemcomitans [Slots, 1982].