PENCUCIAN ERITROSIT DARAH MANUSIA UNTUK MENGOPTIMALKAN PERTUMBUHAN Haemophilus influenzae PADA MEDIA AGAR COKLAT DARI DARAH MANUSIA

Abstract

Background: Sheep Blood Chocolate Agar (SCA) is the standard media for microbiological examination off some fastidious bacteria, including H.influenzae. In many developing countries, the preparation of SBCA is quite difficult, so Human Blood Chocolate Agar (HCA) is used for alternative media although it performance is not as well as the standard SCA. This study aims to find ways to modify the HCA so that it has the culture performance as well or better than the SCA. Methods: This research uses a True-Experimental Post Test Only design in 11 strains of H.influenzae. Bacteria suspension was diluted with a ratio 1:5000 with 0.5 McF density. The whole strains were grown on three types of agar plate in triplo in two ways, first its inoculated in a pure suspension, grown in SCA, HCA and HCAwashed, to assess the colony count, colony diameter and colony characteristics. The second way is spiking the pure suspense into a sputum then it was inoculated in three media (For the sputum spiked suspension, the colony count was not observed). The obtained data were described in graphics, analyzed by Chi-square or Kolmogorov- Smirnov test for the colony count and colony characterictics data, Anova test for colony diameter data, and processed by computer. Result :The colony diameter of H. influenzae pure suspension grown in the SCA, after 24 h incubation showed greater results when compared with HCA and HCAwashed (respectively p = 0.000). At 48 h incubatioan, the diameter of HCA-washed was as big as SCA dan bigger than HCA. The number and characteristics of bacteria colonies from pure suspensions of H. influenzae did not show significant differences in all types of order media. At H.influenzae sputum spike suspension plant on all three media, obtained the colony diameter of 24 and 48 h incubation at HCAwashed is greater when compared with SCA and HCA, and also statistically significant. Conclusion: HCA standard without modification is not recommended for culturing H.influenzae, whereas HCA-washed is recommended as a replacement media SCA. Keywords: Haemophilus influenzae, culture, washing, human blood