Tests demonstrated that volatile chemicals emitted from Enterobacter agglomerans, a bacterium that has been isolated from adults as well as fruit infested with larvae of the Caribbean fruit fly, Anastrepha suspensa (Loew) and other pest fruit flies, are attractive to female A. suspensa in laboratory bioassays. 3-Methyl-1-butanol and ammonia were identified as the two primary volatile chemicals released from active cultures of E. agglomerans. No 3-methyl-1-butanol and little ammonia (16.0 μg/hr) are released from sterile tryptic soy agar plates. E. agglomerans-inoculated tryptic soy agar plates, however, released an average of 1.5 ± 0.53 μg/hr 3-methyl-1-butanol and 332.9 ± 239.16 μg/hr ammonia after 24 hr of growth. 3-Methyl-1-butanol lures were formulated in a membrane-based system to provide a constant release rate of synthetic chemical. Release rates ranged from 0.046 ± 0.007 to 12.16 ± 2.76 μg/hr. In laboratory tests, equal numbers of females were captured in response to ammonium carbonate lures that released ammonia at the rate of 100 μg/hr and to 3-methyl-1-butanol lures that released 12.16 ± 2.756 μg/hr of synthetic material. The combination of the two lures was more attractive than ammonia alone. Availability of lures formulated for a range of 3-methyl-1-butanol release rates will facilitate field tests of this putative microbial attractant and may lead to a better understanding of the role of bacteria in the ecology of pest fruit flies.