Non-perforating abomasal lesions occur with a high prevalence in slaughtered dairy cattle. Ante mortem diagnosis is a challenge, but the presence of occult blood in feces is suggested as a diagnostic criterion. The lower detection limit of Hemo-Fec® (Med-Kjemi, Asker, Norway) and Hemoccult II® SENSA® (Beckman Coulter, Brea, California, USA) for fecal occult blood were estimated. The Hemo-Fec® and Hemoccult II® SENSA® could detect 1-2 mL and 2-4.5 mL of blood in 1000 g of feces, respectively. Therefore, the Hemo-Fec® test was selected to access hemoglobin degradation in the rumen to establish if blood from outside the gastrointestinal tract could result in false-positive tests and an observational study to estimate the diagnostic sensitivity and specificity. Rumen microbiota did not degrade hemoglobin in a 1% blood concentration in vitro during 48 h of fermentation. The Hemo-Fec® test was only able to detect cattle with ≥4 acute lesions (diagnostic sensitivity: 0.40 [95% confidence interval (95% CI): 0.32-0.48] and ≥4 chronic lesions (sensitivity: 0.44 [95% CI: 0.35-0.52]). The Hemo-Fec® test had no diagnostic potential to detect superficial erosions or scar tissue in abomasa. Furthermore, the specificity was 0.71 [95% CI: 0.68-0.75%], and a positive test is thus not equivalent with abomasal lesions in cattle.