The inhibition of acetoclastic methanogenesis in crude oil- and creosote-contaminated groundwater was studied. The crude oil and water-soluble components of creosote contributed to the inhibition of acetoclastic methanogens. Acetoclastic methanogenesis was much more susceptible to the toxic inhibition of crude oil and creosote than either hydrogen- or formate-utilizing methanogenesis. The effect of this toxic inhibition was apparent in the population of the methanogenic trophic groups near nonaqueous crude oil at the Bemidji, MN, site. At a crude oil-contaminated site, numbers of acetoclastic methanogens found close to crude oil were 100 times fewer than those of hydrogen- and formate-utilizing methanogens. In laboratory toxicity assays, crude oil collected from the site inhibited methane production from acetate but not from formate or hydrogen. Toxicity assays with aqueous creosote extract completely inhibited acetate utilization over the range of tested dilutions but only mildly affected formate and hydrogen utilization. Wastewater reactor studies indicated that this toxicity would result in a decrease in the biodegradation rate of contaminants at sites where toxic compounds are present.