Objectives The objective of this study was to evaluate wild-caught mosquitoes for evidence of hemotropic Mycoplasma species DNA and to determine whether the feline hemoplasmas, Mycoplasma haemofelis (Mhf) and ‘ Candidatus Mycoplasma haemominutum’ (Mhm), can be transmitted by Aedes aegypti mosquitoes in a laboratory setting. Methods Wild-caught mosquito pools (50 mosquitoes per pool, 84 pools) utilized in routine public health department disease surveillance programs were tested for hemotropic Mycoplasma species DNA using PCR with primers designed to amplify all known hemoplasmas. Additionally, mosquitoes were trapped in the vicinity of known feral cat colonies, pooled (50 mosquitoes per pool) and tested (84 pools). Purpose-bred cats housed in a research facility were infected with Mhf or Mhm and then colonized laboratory A aegypti were fed upon the bacteremic cats. After a 7 day incubation period, mosquitoes previously fed on infected cats were allowed to feed again on naive cats, which were monitored for bacteremia for 10 weeks. Results Mycoplasma wenyonii DNA was confirmed in one wild-caught mosquito pool by DNA sequencing. While 7% of cats tested in feral colonies were hemoplasma positive, none of the mosquitoes trapped near colonies were positive. Hemoplasma DNA was amplified from A aegypti by PCR immediately after the infectious blood meal, but DNA was not detected at 7 and 14 days after feeding. Although evidence for uptake of organisms existed, hemoplasma DNA was not amplified from the experimentally infested cats in the 10 week observation period. Conclusions and relevance While wild-caught mosquitoes contained hemoplasma DNA and laboratory reared A aegypti mosquitoes take up hemoplasmas during the blood meal, there was no evidence of biologic transmission in this model.