Water dispensers are commonly used in educational institutions in developing countries. Assessment of factors influencing drinking water quality from this source is important due to its potential to cause waterborne diseases and other health risks. This study aimed to assess microbial contamination and the factors affecting drinking water quality from water dispensers at Walailak University, Southern Thailand. Drinking water samples from 60 water dispensers of different type brands were collected and tested for physicochemical and microbiological parameters by following WHO Drinking Water Quality guidelines. Also evaluated by questionnaires were hygienic standards and maintenance of water dispensers. The physicochemical analysis of all water samples did not exceed the reference values of the drinking water standard; the hardness value in only 13.3% of samples was slightly higher than the reference limit. The microbiological results indicated that total coliform and fecal coliform bacteria were found in 16.67% and 8.33% of total samples, respectively. Escherichia coli was not detected in any of the water samples. The assessment of factors influencing drinking water quality, location and pathogen source around water dispensers, hygienic environment, maintenance of water dispensers had a significant effect on drinking water quality.