The geographic range of arthropod vectors, including fleas, ticks, and mosquitoes is expanding worldwide and is influenced by many factors. This expansion increases the risk of contracting vector-borne diseases (VBDs), posing a significant public health threat to both humans and animals, which serve as common reservoirs for filariid nematodes, protozoans, and rickettsial agents. Feline vector-borne diseases (FVBDs), spread by both domesticated and free-roaming cat populations, are often misunderstood and overlooked. The aim of this pilot study is investigate the prevalence of FVBDs in free-roaming cat populations in north-central and south Florida by working with shelter medicine groups and to use feline ear tips discarded during TNR clinics. Ear tip samples (n =) were collected from a shelter in Marion County, Florida, and a shelter in Miami-Dade County, Florida, from September 2024 to June 2025. These ear tips will be evaluated by qPCR for Dirofilaria spp. and Trypanosoma cruzi and Rickettsial species, all important to feline, wildlife and human health. Results will identify the presence and spread of FVBD as the range of free-roaming cats often overlaps with humans and wildlife, so they may prove to be novel and practical sentinels for public and animal health threats. Additionally, this work will highlight and encourage further research in areas where free-roaming cats, humans and wildlife live in shared and changing environments.