Although gingivitis is initiated by plaque and plaque removal controls gingivitis, gingival irrigation with water has been shown to reduce gingivitis without reducing plaque. This study attempted to explore possible mechanisms involved in the treatment of gingivitis by water irrigation. Patients (n = 125) with more than 20 teeth, less than 4 sites with probing depth (PD) deeper than 6 mm, bleeding on probing (BOP) frequency of 30% or higher, and no systemic disease were randomized to one of four treatment groups: toothbrushing alone (brush), toothbrushing plus chlorhexidine 0.12% rinse 2x/day (CHX), toothbrushing plus water irrigation 1x/day (irr+H2O), or toothbrushing plus chlorhexidine 0.04% irrigation 1x/day (irr+CHX). Six sites/tooth were examined at baseline, and at 3 and 6 months for BOP and PD using an automated probe, and for gingival index (GI) and plaque index (PI) by standard means. A prophylaxis and oral hygiene instructions were provided after baseline and 6 month measurements. Subgingival microbial samples and crevicular fluid (GCF) were collected from 2 teeth/subject at each time point. Microbial samples were processed for anaerobic culture and the predominant cultivable flora was determined. CHX and irr+CHX had a 30 to 35% decrease in mean PI, while brush and irr+H2O had only a 12 to 16% decrease. BOP was reduced by 14% in the brush group and 23 to 24% in the other groups. GI was significantly correlated with PI in the brush, CHX, and irr+CHX groups, but not in the irr+H2O group. Prevotella intermedia was significantly reduced in both irrigation groups, but not CHX or brush groups.(ABSTRACT TRUNCATED AT 250 WORDS)