Presence of pathogen especially Salmonella spp in the Betel leaves suspended export of Betel leaf in Europe. Bangladesh has a subtropical monsoon so the present study was undertaken to determine Microbial loads of Betel leaf on the basis of seasonal variation (rainy and winter season). A total of 50 Betel leaf samples were collected from five sources (betel field, transport, whole seller, local shop, betel leaf washing water used in local shop Highest TVC (total viable count) were counted from local shop sample (5.3X 105CFU/ml) and the lowest TVC was found from field sample (2.5X 103 CFU/ml). This study results showed that during rainy season (July-October) TVC count was higher than winter season (November-February). From this study 10 genera of bacteria, were isolated from betel leaf such as E.coli, Vibrio spp, Bacillus spp ,Pseudomonas spp, Klebsiella spp, Salmonella spp, Shigella spp, Staphylococcus spp, Enterococcus spp and Proteus spp) and 5 genera of fungus (e.g. Aspergillus spp , Fusarium spp, Rhizopus spp, Zygosaccharomyces spp and Rhizoctonia spp )were isolated. Out of 184 isolates we found the following percentage of isolated microorganisms: 17.9% in betel leaf field, 19.5% in Transport, 19.5% in wholesaler, 28.8% in local shop and 14.3% in betel leaf washing water from local shop. Antibiotic sensitivity test showed that all of the isolates were resistant to Bacitracin, Penicillin, Vancomycin, Erythromycin and against other 5 antibiotics (Azithromycin, Gentamycin, Cephalexin, Ciprofloxacin and Chloramphenicol) isolates showed Resistant, Moderate and Sensitive Results.Data of this study suggest that Betel leaves from different source could harbor multidrug resistant bacteria specially Salmonella spp which underscore the need of implementation of hygienic practices during production, harvesting, transportation, storage, selling and preparation of Betel leaves to safeguard public health.