Dairy cows may serve as asymptomatic carriers of Salmonella. The potential for herd carrier status increases with herd size, and Salmonella shedding may be triggered by stresses placed on the animals. The scope of the current study is to determine the effects lactation may have on Salmonella genotypic diversity among detected serotypes. Fecal samples were collected on two sampling dates from 60 nonlactating and 60 lactating Holstein cows. No serotype was predominant over the two collection dates, although Salmonella Albany, Salmonella Anatum, Salmonella Newport, and Salmonella Senftenberg were detected in relatively high numbers. Twenty-three genotypes were detected on the first date and 27 on the second date. The greatest genotypic diversity was seen among Salmonella Newport and Salmonella Senftenberg, with five and nine genotypes, respectively. The presence of multiple serotypes and genotypes in the herd suggests multiple contamination sources. However, there was no conclusive effect of lactation status of the cows on Salmonella genotypic shedding.