Neutrophil gelatinase-associated lipocalin (NGAL) is a neutrophil-derived protein whose concentration increases in plasma and urine with ongoing renal damage. Urinary leucocytes can be a potential source of urinary NGAL. The aim of this study is to investigate the effects of urinary neutrophil count and other urinary parameters on urinary NGAL values in urine with negative culture. Urinalysis, urine culture, and determination of urinary NGAL were performed on 33 clinically healthy nonproteinuric dogs with negative urinoculture. The median uNGAL concentration in dogs in this study population was 9.74 ng/mL (IQR 1.93–25.43 ng/mL). In samples with WBCs > 5 hpf (mean 15.9, 6–50 leucocytes/hpf), median uNGAL value was significantly higher than that in samples with WBCs < 5 hpf (mean 0.9, 0–3 leucocytes/hpf), (4.96 pg/mL (0.29–11.34) and 23.65 pg/mL (20.04–29.80), resp.; P=0.0053). The severity of urinary pyuria and the UPC value were correlated with uNGAL concentration. The results of our study show that urinary NGAL concentration is correlated with WBCs number in urinary sediment of dogs with negative urinoculture. The present study suggests that noninfectious pyuria is significantly correlated with urinary NGAL values and might influence uNGAL values.