Dandruff is a common problem in both teens and adults. This study is to evaluate the role of bacteria and fungi associated with dandruff and seborrheic dermatitis. Malassezia furfur (70%) was the predominant isolate, followed by Malassezia globosa (30%) which included mixed infection (15%) of both M. furfur and M. globosa together adding as the significant causative agents (p < 0.00001) as compared to healthy teens. A qualitative in-vitro susceptibility study was performed with Ketoconazole which showed good in-vitro anti-Malassezia activity with a greater inhibitory zone, and similar anti-Malassezia activity was shown by tea tree oil and 1% selenium sulfide. A follow-up study was performed after treatment with 1% selenium sulfide shampoo and showed 92.5% efficiency which suggests a possible solution for dandruff and seborrheic dermatitis.