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This study was conducted to investigate the effects of hot melt extrusion (HME) nano-iron as an alternative for the common ferrous sulfate on iron (Fe) bioavailability, growth performance, nutrient digestibility, intestinal morphology, and intestinal microbiota of weanling pigs. A total of 200 piglets (Landrace × Yorkshire × Duroc) were randomly allotted to seven treatments on the basis of initial body weight (BW) and sex. Treatments were the INO100 (100 ppm Fe as FeSO4), HME-Fe levels (50, 75, and 100 ppm nano-Fe as FeSO4). ORG100 (100 ppm Fe as iron methionine). In phase 1, the HME50 pigs showed the lowest Fe content in feed and feces. Plasma Fe concentration was increased in HME100 and ORG100 pigs. In phase 2, there were significantly lower concentration of Fe in feed and feces of HME50 pigs (p < 0.01). A lower Fe concentration in the plasma and liver were observed in HME50 pigs compared with HME100 pigs. Concentration of red blood cell (RBC) was the lowest (p < 0.01) for HME50 pigs. During phase 2, the HME100, HME75, and ORG100 pigs showed a higher RBC and hemoglobin values compared with HME50 pigs. Digestibility of gross energy (GE) and crude protein (CP) were significantly higher in HME100 pigs compared with HME50 pigs. There was an increased (p < 0.01) villus height in the duodenum and jejunum of HME100 pigs compared with HME50 pigs. It is concluded that dietary Fe does not improve growth performance of weanling pigs; however, increasing the dietary iron concentration in weanling piglets increased the RBC and hemoglobin. In addition, the potential ability of HME to be used at a lower level (HME75) was observed.

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