Heat stress negatively affects milk quality altering its nutritive value and cheese making properties. This study aimed at assessing the impact of seasonal microclimatic conditions on milk quality of Friesian cows. The study was carried out in a dairy farm from June 2013 to May 2014 at Beni-Suef province, Egypt. Inside the barn daily ambient temperature and relative humidity were recorded and used to calculate the daily maximum temperature-humidity index (mxTHI), which was used as indicator of the degree of heat stress. The study was carried out in three periods according to the temperature-humidity index (THI) recorded: from June 2013 to September 2013 (mxTHI>78), from October 2013 to November 2013 (mxTHI 72–78) and from December 2013 to April 2014 (mxTHI<72). Eighty Friesian lactating dairy cows were monitored in each period. The three groups of cows were balanced for days in milk and parity. Milk quality data referred to somatic cell count, total coliform count (TCC), faecal coliform count (FCC), Escherichia coli count, percentage of E. coli, and Staphylococcus aureus, percentage of fat, protein, lactose, total solid and solid non-fat. Increasing THI was associated with a significant decrease in all milk main components. An increase of TCC, FCC, and E. coli count from mxTHI<72 to mxTHI>78 was observed. In addition, the isolation rate of both S. aureus and E. coli increased when the mxTHI increased. The results of this study show the seriousness of the negative effects of hot conditions on milk composition and mammary gland pathogens. These facts warrant the importance of adopting mitigation strategies to alleviate negative consequences of heat stress in dairy cows and for limiting related economic losses.