The Egyptian fruit bat, Rousettus aegyptiacus, was recently identified as a major natural reservoir host for Marburg virus (MARV). Experimental inoculation of R. aegyptiacus from the laboratory-breeding colony with MARV confirmed that this bat species is indeed the natural reservoir host of the virus. However, the mechanisms of bat-to-bat transmission of MARV among R. aegyptiacus remain unknown. Therefore, the objective of this study was to determine whether bat-to-bat transmission of MARV can occur in a controlled laboratory environment through the direct, indirect and/or airborne routes. This study used a total of 38 juvenile bats: 12 inoculated donor (ID) bats, 24 naïve contact (NC) bats and 2 negative control (NEG CO) bats. The ID bats were inoculated with MARV, the NC bats received no inoculation and the NEG CO bats were inoculated with media. The ID and NC bats were housed in partitioned cages, separated by wire mesh or solid metal partitions. Blood, urine, and oral and rectal swabs were collected from bats daily from 0 through 25 dpi and then weekly through 56 dpi. These specimens were used to monitor viremia and virus shedding by Q-RT-PCR and virus isolation, and to monitor the MARV IgG antibody response. This study identified multiple routes of MARV shedding in the ID bats and evidence of oral exposure in NC bats. Further transmission studies will be necessary to better characterize bat-to-bat transmission of MARV in a controlled laboratory environment.