Adipose tissue macrophages (ATM) infiltration is a key characteristic of the pro-inflammatory state during obesity in humans and rodents. However, it is unknown if ATM infiltration is associated with periparturient disorders of dairy cows. The aim of this study was to quantify the number of ATM in subcutaneous adipose tissue (AT) during the transition period of dairy cows. Five high BCS (BCS ≥ 3.75) and 4 medium BCS (2.75 ≤ BCS ≤ 3.5) multiparous Holstein cows were used. Blood samples and tailhead AT samples were obtained at 3 time points relative to the calving date: far-off (−27 ± 2.2 d), close-up (−10 ± 1.5 d) and fresh (9 ± 0.3 d). AT samples were fixed in 4% formaldehyde or frozen in liquid nitrogen. Number of CD172a⁺ cells (marker for ATM) was determined by immunohistochemistry. Adipose gene expression of CCL2, CCL22, CD68, and SIRPA was determined by RT-qPCR. The effect of BCS, NEFA, BHB, period, disease (as recorded by the farm personnel) and gene expression on the proportion of CD172a⁺ cells was determined using a general linear mixed model with period as repeated observation within subject cow. BCS, NEFA and BHB had no effect on the proportion of CD172a⁺. The proportion of CD172a⁺ cells was greater in AT samples from fresh diseased cows (6.14 ± 0.84%) compared with all other samples (far-off healthy, 0.86 ± 0.68%, P < 0.01; far-off diseased, 2.49 ± 0.84%, P < 0.05; close-up healthy, 2.34 ± 0.68%, P = 0.06; close-up diseased, 2.39 ± 0.84%, P < 0.05; fresh healthy, 0.86 ± 0.68%, P < 0.01). Similarly, the proportion of CD172a⁺ cells was associated with an interaction between period and the expression of CCL2 (P < 0.01), CCL22 (P < 0.01) and CD68 (P < 0.05). During the fresh period, adipose gene expression of CCL2, CCL22, and CD68 was positively associated with the proportion of CD172a⁺ cells. The results of this study indicate that ATM infiltration of the subcutaneous AT is associated with diseases in early lactation dairy cows. However, it is still unknown whether ATM infiltration is the cause or consequence of periparturient diseases.