One of the major causes of mortality in poultry is urolithiasis. Although two of the major causes of urolithiasis have been defined, active and preventative treatments for this disease have not been extensively field tested. Previous research demonstrated that a diet acidified with NH4Cl was effective in preventing the formation of uroliths in 16-wk-old pullets. In the present study, experiments were designed to evaluate the effects of dietary acidification and alkalinization on preformed uroliths in mature laying hens. Urolithiasis was induced by feeding excess dietary Ca (in commercial layer ration) during pullet growout (6 to 18 wk of age) and during the early laying period (18 to 32 wk of age). At 32 wk of age birds were randomly divided into three diet treatment groups: a normal layer ration group, an acidified layer ration group (1% NH4Cl added); and an alkalinized layer ration group (1% NaHCO3 added). Each group received the respective diets until 52 wk of age, when the experiment was terminated. Blood gas analysis at 44 wk of age demonstrated that hens fed the acidified diet had significantly higher blood H+ concentrations, lower blood bicarbonate concentrations, and lower total blood carbon dioxide levels than hens fed the alkalinized layer ration. At 52 wk of age, hens fed the acidified layer ration had higher urinary H+ concentrations, higher blood H+ concentrations, and lower blood HCO3 and total CO2 concentrations than hens fed the normal and alkalinized layer rations. The acidified layer ration group had no uroliths present in their ureters (0% urolithiasis), whereas the normal and alkalinized layer ration groups had 8.3 and 13.1% incidences of urolithiasis.(ABSTRACT TRUNCATED AT 250 WORDS)