Untreated abattoir effluent constitutes potential reservoir for transmission of pathogenic strains of multiple antibiotic-resistant bacteria by pollution of surface and ground water sources. This study was carried out to determine the antibiotic resistance and extended spectrum β-lactamase (ESBL) production profiles of Gram-negative bacteria isolated from effluent collected from Lafenwa municipal abattoir and its receiving surface water, Ogun River, in Abeokuta, Ogun state, Nigeria. Twelve effluent and 18 water samples were collected for this study. Total heterotrophic and coliform counts were estimated, bacterial identification was performed using standard culture-based procedures, whilst antibiotic resistance profiles of isolated bacteria against five antibiotics (ceftazidime, cefpodoxime, cefotaxime, ertapenem and amoxicillin-clavulanate) and detection of ESBLs were done using disk diffusion and double-disc synergy tests. A total of 54 Gram-negative bacteria were isolated, including Salmonella spp. (9), Escherichia coli (15), Klebsiella spp. (7), Shigella spp. (5), Pseudomonas spp. (12) and Enterobacter spp. (6). Both Enterobacteriaceae and Pseudomonas isolates (31% and 66.6%, respectively) were resistant to all selected antibiotics except ertapenem (98% susceptibility). Overall, 77% isolates had multiple antibiotic resistance index (MARI) values, but none of the antibiotic-resistant isolates showed evidence of ESBL production. The presence of multiple antibiotic-resistant isolates in the effluent and receiving water of Lafenwa abattoir suggests a major risk to public health and food safety. Current methods of waste disposal at the abattoir are unacceptable and greatly reduce the qualities of the processed meat and contaminate the environment. There is a need for improved abattoir waste management and water treatment strategies.