Non-invasive methods of skin condition assessment include, among others, the evaluation of transepidermal water loss (TEWL). The aim of the study was to examine whether TEWL values depend on horse breed. The study was conducted on four breeds: “Felin” ponies (FP) (n=16), Polish koniks (PK) (n=15), Polish cold-blooded horses (PcbH) (n=11) and Wielkopolska horses (WH) (n=12). It was found that horse breed influences TEWL values. In the neck region, statistically significant differences were found between PK and FP (p=0.006), and PK and WH (p=0.0005). In the lumbar region, there were statistically significant differences between FP and PK (p=0.0009), FP and PcbH (p=0.0016) as well as between PK and WH (p=0.000037), and PcbH and WH (p=0.0006). In the inguinal region statistically significant differences were found between FP and PK (p=0.0003), FP and PcbH (p=0.0005), PK and WH (p=0.009) and PcbH and WH (p=0.006). In the lip region statistically significant differences were observed between FP and PK (p=0.013) as well as between PK and PcbH (p=0.029) and PK and WH (p=0.009). In the examination of TEWL animal breed should be taken into consideration. The non-significant differences found in three of the examined body regions may suggest that these regions are the most adequate for TEWL assessment.