Herpes simplex virus, cytomegalovirus and Epstein-Barr virus infections are prevalent among HIV-1 infected persons. The relationships between salivary shedding of these herpesviruses have not been characterized. Salivary samples were collected on a median of 61 consecutive days from 41 HIV-1 seropositive persons and tested for HSV-1, HSV-2, CMV and EBV. HSV was detected on 5%, CMV on 19% and EBV on 71% of the days of sampling. HSV shedding was not related to CMV or EBV shedding rates. Persons with EBV shedding rates >40% had CMV DNA detected in their saliva significantly more often than those with EBV shedding rates <or=40% (P=0.008). The odds of detecting CMV were greater on days with HSV (OR 2.5, 95% CI 1.4, 4.4) or EBV (OR 3.8, 95% CI 1.9, 7.9) shedding. No significant associations between HSV shedding rates and CD4 count, plasma HIV-1 RNA or HAART were observed. Increasing plasma HIV-1 RNA was associated with greater frequency (P=0.01) and quantity (P<0.001) of EBV shedding. Among persons not receiving HAART, CD4 counts >200 cells/mm(3) were associated with lower frequency (P=0.02) and quantity (P=0.03) of CMV compared with CD4 counts <or=200 cells/mm(3). These data suggest that separate factors influence mucosal shedding of each of the three classes of herpesviruses but that virological interactions between the pathogens also exist.