AIMS: Bilateral optic neuropathy in Dar-es-Salaam, Tanzania was first reported as an epidemic in 1988. Now argued to be endemic in 2010, the aetiology remains unclear. The authors investigated the hypothesis that low folate and vitamin B₁₂ status are associated with optic neuropathy, and also sought to investigate whether mercury, commonly used drugs, dietary factors and indoor pollution may also be risk factors. METHODS: 57 cases and 102 controls were recruited from two tertiary referral centres in Dar-es-Salaam. Data were collected on demographic characteristics, diet, medication history and HIV status. Folate and vitamin B₁₂ (holo-transcobalamin) were measured in stored serum samples. Exposure to mercury was assessed from concentrations in random urine samples. RESULTS: Cooking indoors more than twice per week (OR 54.48 (95% CI 9.30 to 319.10)) and indoor use of charcoal or firewood (OR 21.20 (95% CI 2.51 to 179.36)) increased the risk of optic neuropathy. Risk was reduced in those with a higher folate status (highest versus lowest quartile OR=0.11 (95% CI 0.02 to 0.51)) and higher protein intakes (OR=0.84 (95% CI 0.72 to 0.96). No association was found with mercury exposure or any common drug or food commodity. CONCLUSION: This study presents the first direct evidence of low folate status and indoor pollution in the aetiology of endemic bilateral optic neuropathy in Tanzania.