The objective of this study is to evaluate the seminal fluid for the presence of pathogenic microbes and their effect on the fertility of infertile male patients from bilaspur. For achieving a desired interpretation I have collected 60 semen samples from infertile and regular male patients. After collection following procedures like streaking, morphological study and biochemical tests are done for identifying pathogenic microbes present in semen samples. After studying every sample I came to the conclusion that spp. Of Staphylococcus, E. coli, Enterococcus, Candida spp. etc are the most common pathogens present in semen. The result also correlates the transmission of microbes in some patients from their female partners through vagina (vaginal micro flora). Due to the presence of these microbes the male patients are facing low sperm quantity as well as quality.