Arthropod vectors play a crucial role in the transmission of many debilitating infections, causing significant morbidity and mortality globally. Despite the economic significance of arthropods to public health, public knowledge on vector biology, ecology and taxonomic status remains anecdotal and largely unexplored. The present study surveyed knowledge gaps regarding the biology and ecology of arthropod vectors in communities of Botswana, across all districts. Results showed that communities are largely aware of individual arthropod vectors; however, their ‘potential contribution’ in disease transmission in humans, livestock and wildlife could not be fully attested. As such, their knowledge was largely limited with regards to some aspects of vector biology, ecology and control. Communities were strongly concerned about the burden of mosquitoes, cockroaches, flies and ticks, with the least concerns about fleas, bedbugs and lice, although the same communities did not know of specific diseases potentially vectored by these arthropods. Knowledge on arthropod vector control was mainly limited to synthetic chemical pesticides for most respondents, regardless of their location. The limited knowledge on potentially pathogen-incriminated arthropod vectors reported here has large implications for bridging knowledge gaps on the bio-ecology of these vectors countrywide. This is potentially useful in reducing the local burden of associated diseases and preventing the risk of emerging and re-emerging infectious diseases under global change.