Water quality is critical for poultry farming. This study assessed the physical, chemical and microbiological quality of drinking water in small-layer farms in Southern Mozambique and identified potential risk factors for total coliform (TC) and Escherichia coli contamination of drinking water. In 20 farms, 57 samples were collected and examined for pH, nitrate content (NC), nitrite level (NL) and total hardness contents (TH). Furthermore, TC and E. coli growth were assessed at 37 °C. One hundred per cent of the drinking water was of acceptable quality in terms of pH (6.5-8.5), NC (50 mg/L) and NL (3 mg/L). Total hardness contents exceeded the recommended standard in 37.5% of borehole water samples and 91.7% of tap water samples, respectively. Total coliform and E. coli were found in 40% and 15% of water samples. Tap water samples had the greatest contamination, with TC and E. coli levels of 41.7% and 16.7%, respectively. Although not statistically significant, sampling from the beginning of the nipple line (p = 0.101, OR = 7.357, 95% confidence interval [CI]: 0.678-79.886) and not cleaning the rearing equipment regularly (p = 0.098, OR = 3.966, 95% CI: 0.766-20.280) were factors affecting the TC growth. Sampling from the tank water source (p = 0.001, OR = 0.005, 95% CI: 0.000-0.121) and borehole water source (OR = 13 585) and not cleaning the equipment consistently (p = 0.073, OR = 9.682, 95% CI: 0.810-115.68) were all factors affecting E. coli growth. It is concluded that the TH and microbiological quality of the drinking water of the study region are inadequate. Regular water quality assessments should be incorporated into Mozambican layer farm management to limit the potential for health concerns, and farmers should thoroughly clean and disinfect their rearing equipment. CONTRIBUTION: We should incorporate regular water quality assessments into Mozambican layer farm management to limit the potential for health concerns, and farmers should thoroughly clean and disinfect their rearing equipment