A total of 327 root samples collected from horticultural and agronomic crops, and weeds associated with these crops in Florida crop production regions, were examined. Isozyme phenotypes, esterase (EST) and malate dehydrogenase (MDH) phenotypes were used to characterise and identify the species of Meloidogyne present in these samples. At least 26 females from each sample were examined using polyacrylamide gel electrophoresis. Sixteen major bands of EST activity were found, corresponding to 12 phenotypes. A species-specific EST phenotype was consistently associated with each of the Meloidogyne spp. identified: Meloidogyne arenaria, M. floridensis, M. graminicola, M. hapla, M. incognita, M. javanica, M. mayaguensis and M. partityla. Two unique EST phenotypes, which have not been described, were found associated with three unidentified nematode populations. Five bands of MDH activity and four phenotypes were also found among the populations. During this study, new host records were determined for M. floridensis and M. mayaguensis, two recently reported Meloidogyne spp. in Florida. Plant species found as new hosts of M. floridensis were Cucumis sativus, Phaseolus sp., Solanum melongena and one weed species, Emilia sonchifolia, and the M. incognita-resistant peach rootstock 'Flordaguard'. New hosts for M. mayaguensis were Carica papaya, Capsicum annuum var. Longum, and six weed plants, including Eclipta prostrata, Fatoua villosa, Panicum sp., Poinsettia cyathophora, Solanum americanum and also one unidentified weed species belonging to the family Acanthaceae. To our knowledge, Florida is the only geographic area of North America where M. floridensis and M. mayaguensis have been detected.