The predominant microorganisms that cause subclinical and clinical intramammary infections (IMI) can vary with environmental conditions, personnel, and management practices. However, limited information exists regarding the types of organisms isolated from subclinical IMI of dairy cattle in the southeast US. To address this limitation, members of the Southeast Quality Milk Initiative identified 30 herds in KY (n = 9), MS (n = 3), TN (n = 9), and VA (n = 9) that were participating in Dairy Herd Improvement programs and willing to participate. The top 10–20 cows with the highest SCC according to the most recent DHI test were selected for sampling at each farm visit (2–3 times per year, Jan 2016 to Jan 2018). Quarter milk samples were collected aseptically and stored at −20C until cultured. Culture and organism identification were performed as outlined by NMC. Overall, 1,847 cows and 5,710 quarters were sampled. Of these, 647 (11.3%) were considered contaminated with either bacillus or 3+ colonies of different morphology and 2411 (42.2%) exhibited no growth. After removing these samples, 2,652 quarter samples contained 1 (95%) or 2 (5%) bacterial isolates. The dominant organisms isolated included coagulase-negative staphylococci (CNS; 38.9%), Staphylococcus aureus (15.6%), Streptococcus dysgalactiae (7.6%), Esculin positive streptococci including Streptococcus uberis (7.3%) and Escherichia coli (5.3%). The following organisms each accounted for 1 to 3.5% of the isolates: Staphylococcus hyicus, Serratia, Corynebacterium, Klebsiella, Enterococcus, yeast, and Aerococcus species. CNS and S. aureus were the 2 most isolated organisms on 16 of 30 farms. Closer examination of a subset of CNS isolates, revealed 41 of 61 (67%) were Staphylococcus chromogenes. This suggests that S. chromogenes may be a more frequent cause of subclinical IMI with high SCC than S. aureus. Staphylococcal species, including S. aureus, represent the predominant isolates from high SCC cows in the southeast and strategies to prevent or minimize new infections with these organisms should be a part of mastitis control programs in the southeast US.