This experiment studied the effects of dietary protein sources and levels on the gut health of piglets, pH value, and concentrations of microbial metabolites (ammonia-N, volatile fatty acids [VFA], and polyamines) in the distal colonic and proximal colonic digesta of piglets weaned at 21 d of age. A total of 150 early-weaned piglets were allotted randomly to 5 diets: 1) control diet (CT; 17% CP), 2) CT formulated with more soy protein concentrate (SPC19; 19% CP), 3) more fish meal (FM19; 19% CP), 4) CT formulated with more soy protein concentrate (SPC23; 23% CP), and 5) more fish meal (FM23; 23%CP). Results showed high protein level increased fecal score (P < 0.05), but different protein sources did not (P > 0.05). The pH value and ammonia-N concentration of digesta in the proximal and distal colon of FM23 were significantly higher (P < 0.05) than those of CT. Acetic acid, propionic acid, butyric acid and valeric acid concentrations in the proximal colon of FM23 exceeded those of CT, SPC19, and FM19 (P < 0.05); however, isobutyric acid and isovaleric acid were not affected (P > 0.05). Histamine and spermidine concentrations of FM23 were higher than those of other treatments (P < 0.05). Propionic acid and butyric acid concentrations in the distal colon were higher of FM23 than of FM19 (P < 0.05); putrescine, histamine and spermidine were higher of FM23 than of LP and FM19 (P < 0.05). It was concluded that high dietary CP content increased microbial metabolites (ammonia-N, histamine, putrescine) in colonic digesta and aggravated piglets' diarrhea.