This longitudinal study of 194 very-low birthweight (VLBW) and 184 normal birthweight (NBW) infants hypothesized that the causal pathway between birth group (VLBW or NBW) and mutans streptococci (MS) acquisition (presence) at 18-20 months is mediated by biological, behavioral, and caregiver MS levels. Biological (number of teeth at 8 and 18-20 months and enamel hypoplasia) and behavioral (brushing/cleaning, sweet snacks, breastfeeding, and dental access) factors were assessed using dental examinations and caregiver questionnaire responses at 8 and 18-20 months. Infant MS acquisition and caregiver MS levels were assessed from saliva and plaque samples collected at 8 and 18-20 months. Structural equation modeling evaluated the causal pathway with latent variables for biology and behavior. Mutans streptococci presence was similar between birth groups at 18-20 months (40% in VLBW infants and 49% in NBW infants), but was significantly higher for NBW infants at 8 months. Increased number of teeth at 8 and 18-20 months was associated with biological risk. Infants whose caregivers had a 1-point higher score on MS had a significantly (1.5) higher odds of MS presence. Caregiver behavior was not associated with MS presence. Early-intervention efforts should focus on delaying initial acquisition and improving caregiver awareness of taking care of erupting primary teeth.