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After collection of the fasted resting breath samples, 3 small meals without isotope were offered every 10-min to induce fed-state in the pigs. A priming dose of 1.75 constant dose (3.5 mg/kg BW) was given once along with the constant dose at meal 6 (Moehn et al., 2003). A constant dose of 2 mg/kg BW (Levesque et al., 2011) of the stable isotope L-[1-13C]-Phe (99%; Cambridge Isotope Laboratories, Inc. Saint-Laurent, QC) was given orally every 25-min between meals 6 and 13, inclusive. All pigs consumed their meals prior to the collection of the corresponding breath samples. The mechanism of breath collection was that air from the chambers was drawn by a rotary vane vacuum pump through a series of drierite-filled columns to the CO2 analyser (Qubit Model S155, Qubit Systems Inc.) and the gas switcher. From the gas switcher, expired air was pushed through midget bubblers, which contained 8 mL of 1 mol/L sodium hydroxide (NaOH) solution. The purpose of the NaOH solution was to trap any CO2 released by the pigs from the chambers for the subsequent 13CO2 enrichment analysis (Shoveller et al., 2017). Breath samples were then stored in an air-tight serum tube and kept at room temperature until further analysis.

In comparison with the SID value of Met from Exp. 1, the MA of Met in the PD-BSFL meal was found to be 37.1 percentage units lower (SID = 90.4% versus MA = 53.3%). The lower MA compared with SID value is expected. However, the magnitude between the two was quite surprising as the two studies were conducted using the same PD-BSFL meal without any further processing, barrows of similar initial BW, age, genetics, and similar ingredient composition in the experimental diets. A potential explanation could be that during the processing of the PD-BSFL meal (i.e., oven drying from wet larvae to dry-powdered meal), Met was exposed to heat and oxidized to Met sulfoxide and Met sulfone. These Met forms can be absorbed by the gastrointestinal tract through the same transport mechanism as Met (Higuchi et al., 1982), but they are nutritionally unavailable for the animals (Anderson et al., 1976; Rutherfurd and Moughan, 2008). When analyzing Met content in the digesta samples in Exp. 1, performic acid oxidation followed by the acid hydrolysis method was used. This approach has been reported to overestimate Met content, as the analysis would also account for Met sulfone and sulfoxide (Batterham, 1992; Wang and Parsons, 1998; Rutherfurd and Moughan, 2008). Therefore, it is likely that in Exp. 1, SID value overestimates Met bioavailability due to absorption of unavailable forms of Met.

We would like to thank Douglas Wey and Cuilan Zhu (University of Guelph) for their help with the ileal cannulation surgery, handling and caring of the pigs, and laboratory analysis. This work was funded by Canada First Research Excellence Fund (499127 and 499128) and an in-kind donation from Enviroflight, LLC. 041b061a72