During a 2016 survey of the nematode associates of the native palmetto weevil,Rhynchophorus cruentatus, and the recently introduced West Indian sugarcane weevil,Metamasius hemipterus(Coleoptera: Curculionidae), from southern Florida, a new species ofAcrostichuswas cultured from a single dissectedR. cruentatusfrom Fort Pierce, FL, USA. Morphological and molecular studies showed that it was new to science and it is described herein asA. floridensisn. sp. The new species is characterised by its male tail characters, spicule morphology with rounded manubrium separated from other parts by clear constriction, smoothly ventrally curved blade, slightly dorsally recurved and pointed tip, more or less straight gubernaculum with widely rounded anterior end and a triangular (arrowhead-like) appendage at the distal tip, and the arrangement of male genital papillae, ⟨(v1, v2), v3 / v4, ad, ph, (v5, v6, v7, pd)⟩. In addition toA. floridensisn. sp. and the previously described nematode associates ofR. cruentatus,i.e.,A. rhynchophori,Teratorhabditis palmarumandMononchoidessp., we recovered a putative new species ofDemaniellaand a new association record withRhabditidoides humicolusandDiplogastrellus metamasiusin Homestead, FL. Dissections and subsequent culturing attempts withM. hemipterusrevealed the previously described nematode associates ofCaenorhabditis angariaandD. metamasiusas well as a new association withR. humicolusin Homestead, FL, USA.