Isolation, identification and characterization of microbial contaminants in selected biosafety laboratories in Kenya
Mose, Dennis Nyachae
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Microbial contamination is a constant problem which is often associated with development of all in vitro techniques in biosafety laboratories. The nutrient media in which manipulations are done is a good source of nutrient for microbial growth which results to increased culture mortality. Presence of latent infections can result to variable growth and tissue necrosis. The massive scale under which biotechnology activities are carried out makes the prospect of microbial contamination in the laboratories even though small proportion clearly costly. The objectives of the study will be to assess sources of microbial contaminants in biosafety laboratories, isolate and determine these contaminants based on their morphological and biochemical characteristics, determine resistance bacteria strains and characterize the resistance bacteria strains by molecular techniques. The study will be carried out in the plant tissue culture and plant transformation laboratories at Kenyatta University, KARl Biotechnology Centre and International Livestock Research Institute (ILRI). PDA plates and NA plates will be exposed to air in the laboratories for 30 min after which they will be sealed with parafilm and incubated at 25°C for 72 h and 37°C for 24 - 48 h for PDA and NA respectively. Sterile cotton buds will be used to swab 2 M on laboratory walls, tables and door knobs, lab coats of laboratory staff and biosafety cabinets before and after sterilizing with hypochlorite based disinfectants. The swabs will be inoculated onto the PDA plates and NA plates and then incubated. After visual examination, contaminated plates will be removed from growth rooms. Fungal isolates will be identified using cultural and morphological characters by comparison with standards. Bacterial identification will be carried out using Gram staining, morphology and biochemical reactions in specific media The percentage data of microbes will be transformed using square root method. Statistical analysis will be carried out using ANOVA with GENESTAT Version 6, computer software. Means will be separated using Tukey's Honest Significance Difference at 5% level. RFLP profiles will be scored for absence or presence of bands and the data will be analysed using Gene Alex computer software. Dendograms will be drawn using Neis Genetic Distance with Popgene computer software.