Fecal calprotectin is a noninvasive marker of gut inflammation and has been widely utilized in human gastrointestinal diagnostics. This marker, however, has not been extensively utilized in porcine samples. The aim of this study was to optimize a protocol for the extraction of porcine fecal calprotectin, and to the best of our knowledge this is the first study to be conducted in this regard. Freshly collected swine fecal samples were used in this study. We determined the variability of three commercial ELISA assays in the recovery of porcine fecal calprotectin. We further studied the effect of dilution factor and roller shaker homogenization on the yield of calprotectin from swine fecal samples. Calprotectin recovery was significantly different(p<0.05) across the three commercial assays with MBS033848 having a greater recovery compared to DAEF-012 and calprest. Fecal calprotectin yield increased with an increase in dilution factor with maximum recovery at 1:250. Furthermore, homogenization of fecal sample extracts using a roller shaker for tubes for 30 min led to a 30.75% relative increase in calprotectin yield. Further increase in shaking time(at 60 min) led to a reduced calprotectin recovery. Calprotectin recovery ratio was 130.8% and 101.4% at 30 min and 60 min homogenization respectively. In our conclusion, we observed that various factors affect the recovery of porcine fecal calprotectin, and therefore the researcher should double check certain parameters in regard to the type of kit, the dilution factor and homogenization time if reliable and reproducible results are to be obtained. Results of the present study provide useful information on a non-invasive protocol to veterinarians and researchers in examining and monitoring swine gut healthusing the fecal calprotectin.

ABSTRACT
 Introduction
 Materials and Methods
  1 Fecal sample collection, handling and storage
  2 Assay variability of commercial calprotectin ELISAkits in the recovery of porcine fecal calprotectin
  3 Effect of dilution factor on the recovery of porcinefecal calprotectin
  4 Effect of roller shaking time on porcine fecalcalprotectin yield
  5 Statistical analysis
 Results and Discussion
  1 Assay variability of the selected commercialELISA Kits
  2 Dilution factor
  3 Roller shaker homogenization time and calprotectinyield
 References

• Paul Bogere(Department of Agricultural Convergence Technology, Chonbuk National University)
• Yeon Jae Choi(International Agricultural Development and Cooperation Center, Chonbuk National University)
• Jaeyoung Heo(International Agricultural Development and Cooperation Center, Chonbuk National University) Corresponding author