Supplementary Table 1 Table of F-Ratios for the Effects of Dietary Treatment and Concentration

Supplementary Table 1 Table of f-ratios for the effects of dietary treatment and concentration of α-chymotrypsin, trypsin, α-amylase and α-glucosidase in the tissues and lumen of the gut (pooling regions) using different covariates; gut wet weight (Model 1), food ingested (Model 2), protein ingested (Model 3) and carbohydrate ingested (Model 4). Models only differ in the covariates used. Numbers in parentheses are P values, with significant values (P < 0.05) highlighted in bold.

Model / 1 / 2 / 3 / 4
α-Chymotrypsin - tissues / contents
Diet treatment / 2.93(0.039) /
3.2(0.028) / 3.09(0.032) /
2.35(0.079) / 1.37 (0.259) /
1.82 (0.151) / 2.50(0.066) /
2.4(0.075)
Covariate / 1.25(0.264) /
1.55(0.215) / 0.91(0.342) /
0.88(0.348) / 1.09 (0.300) /
1.13 (0.288) / 0.42(0.516) /
0.36(0.547)
Error df / 73/73 / 73/73 / 73/73 / 73/73
Trypsin – tissues / contents
Diet treatment / 1.89(0.139) /
0.33(0.804) / 1.93(0.132) /
0.19(0.905) / 1.94(0.132) / 0.24(0.870) / 1.79(0.157) /
0.29(0.832)
Covariate / 0.28(0.593) /
0.01(0.920) / 0.27(0.606) /
0.03(0.870) / 0.44(0.508) /
0.29(0589) / 0.06(0.815) /
0.11(0.738)
Error df / 73/73 / 73/73 / 73/73 / 73/73
α-Amylase – tissues / contents
Diet treatment / 9.15(<0.001) /
7.77(<0.001) / 9.42(<0.001) /
7.69(<0.001) / 7.97(<0.001) /
5.08(0.003) / 5.78(0.001) /
5.77(0.001)
Covariate / 0.79(0.374) /
0.01(0.912) / 1.71(0.192)
0.01(0.976) / 2.23(0.136) / 0.03(0.874) / 0.65(0.419) /
0.07(0.797)
Error df / 67/67 / 67/67 / 67/67 / 67/67
α-Glucosidase – tissues / contents
Diet treatment / 1.31(0.278) /
2.04(0.116) / 1.47(0.230) /
1.12(0.347) / 1.64(0.188) /
1.58(0.202) / 1.09(0.359) /
0.72(0.543)
Covariate / 0.43(0.510) /
1.47(0.226) / 0.86(0.354) /
0.47(0.496) / 1.81(0.18) /
0.22(0.64) / 0.05(0.819) /
0.67(0.415)
Error df / 73/73 / 73/73 / 73/73 / 73/73

Supplementary Table 2 Results for enzyme-like activity for each enzyme in the tissues and contents for each region of the gut given diet treatment and time. Significant P values (P < 0.05) are highlighted in bold.

Analysis / Source of variation / df / MS / F / P
Chymotrypsin-tissues
Between subjects / Time / 2 / 32,231.572 / 0.971 / 0.382
Treatment / 3 / 129,606.419 / 3.906 / 0.011
Time*Treatment / 6 / 36,090.921 / 1.088 / 0.374
Locust(Time*Treatment) / 107 / 33,179.624 / 2.602 / < 0.001
Within subjects
Gut region / 4 / 846,767.522 / 66.399 / < 0.001
Gut region * Time / 8 / 20,963.761 / 1.644 / 0.110
Gut region * Treatment / 12 / 41,279.252 / 3.237 / < 0.001
Gut region * Time*Treatment / 24 / 11,222.576 / 0.880 / 0.630
Error / 428 / 12,752.807
Chymotrypsin-contents
Between subjects / Time / 2 / 4,630,479.015 / 0.222 / 0.801
Treatment / 3 / 34,099,244.417 / 1.635 / 0.026
Time*Treatment / 6 / 7,787,545.254 / 0.373 / 0.894
Locust(Time*Treatment) / 107 / 20,856,317.301 / 4.650 / < 0.001
Within subjects
Gut region / 3 / 7.642 x 108 / 170.399 / < 0.001
Gut region * Time / 6 / 2,520,360.048 / 0.562 / 0.761
Gut region * Treatment / 9 / 12,723,727.528 / 2.837 / 0.003
Gut region * Time*Treatment / 18 / 3,989,605.196 / 0.890 / 0.592
Error / 321 / 4,484,924.991
Trypsin –tissues
Between subjects / Time / 2 / 1,118.852 / 1.389 / 0.254
Treatment / 3 / 282.628 / 0.351 / 0.789
Time*Treatment / 6 / 323.606 / 0.402 / 0.876
Locust(Time*Treatment) / 107 / 805.437 / 5.853 / < 0.001
Within subjects
Gut region / 4 / 4,131.524 / 30.024 / < 0.001
Gut region * Time / 8 / 264.195 / 1.920 / 0.055
Gut region * Treatment / 12 / 321.160 / 2.334 / 0.007
Gut region * Time*Treatment / 24 / 82.764 / 0.601 / 0.933
Error / 428 / 137.605
Trypsin –contents
Between subjects / Time / 2 / 587,371.443 / 1.073 / 0.346
Treatment / 3 / 120,008.510 / 0.018 / 0.895
Time*Treatment / 6 / 225,578.129 / 0.412 / 0.870
Locust(Time*Treatment) / 107 / 547,301.165 / 1.568 / 0.001
Within subjects
Gut region / 3 / 13,931,781.507 / 39.905 / < 0.001
Gut region * Time / 6 / 248,788.790 / 0.713 / 0.640
Gut region * Treatment / 9 / 120,021.521 / 0.344 / 0.960
Gut region * Time*Treatment / 18 / 291,339.474 / 0.834 / 0.659
Error / 321 / 349,122.248
Amylase – tissues
Between subjects / Time / 2 / 1.623 x 108 / 2.755 / 0.069
Treatment / 3 / 3.205 x 108 / 5.440 / < 0.001
Time*Treatment / 6 / 16,148,546.844 / 0.274 / 0.948
Locust(Time*Treatment) / 97 / 58,909,239.479 / 3.542 / < 0.001
Within subjects
Gut region / 4 / 7.911 x 108 / 47.567 / < 0.001
Gut region * Time / 8 / 33,322,950.197 / 2.004 / 0.055
Gut region * Treatment / 12 / 17,523,769.906 / 1.054 / 0.399
Gut region * Time*Treatment / 24 / 22,493,422.252 / 1.352 / 0.126
Error / 386 / 16,631,938.846
Amylase – contents
Between subjects / Time / 2 / 4.883 x 109 / 0.425 / 0.655
Treatment / 3 / 5.204 x 1010 / 4.528 / 0.005
Time*Treatment / 6 / 1.418 x 109 / 0.123 / 0.993
Locust(Time*Treatment) / 97 / 1.149 x 1010 / 6.024 / < 0.001
Within subjects
Gut region / 3 / 3.157 x 1011 / 165.503 / < 0.001
Gut region * Time / 6 / 4.260 x 109 / 2.233 / 0.040
Gut region * Treatment / 9 / 5.399 x 109 / 2.830 / 0.003
Gut region * Time*Treatment / 18 / 1.724 x 109 / 0.904 / 0.574
Error / 291 / 1.908 x 109
Glucosidase-tissues
Between subjects / Time / 2 / 288.714 / 0.935 / 0.396
Treatment / 3 / 548.422 / 1.776 / 0.156
Time*Treatment / 6 / 56.332 / 0.182 / 0.981
Locust(Time*Treatment) / 107 / 308.870 / 1.679 / < 0.001
Within subjects
Gut region / 4 / 16,239.728 / 88.300 / < 0.001
Gut region * Time / 8 / 258.624 / 1.406 / 0.192
Gut region * Treatment / 12 / 105.548 / 0.574 / 0.863
Gut region * Time*Treatment / 24 / 175.941 / 0.957 / 0.524
Error / 427 / 183.916
Glucosidase-contents
Between subjects / Time / 2 / 53,106.632 / 0.018 / 0.982
Treatment / 3 / 4,197,725.171 / 1.462 / 0.229
Time*Treatment / 6 / 1,328,818.141 / 0.463 / 0.834
Locust(Time*Treatment) / 107 / 2,870,748.937 / 2.352 / < 0.001
Within subjects
Gut region / 3 / 46,799,110.526 / 38.336 / < 0.001
Gut region * Time / 6 / 1,066,535.415 / 0.874 / 0.514
Gut region * Treatment / 9 / 1,013,898.154 / 0.831 / 0.588
Gut region * Time*Treatment / 18 / 357,292.338 / 0.293 / 0.998
Error / 320 / 1,220,757.387

Supplementary Table 3 Results of ANCOVA for the activity of chymotrypsin-like activity in the tissues and amylase-like activity in the tissues and contents for each region of the gut. Under ‘Analysis’, covariate and P values are those for the homogeneity of slopes. Pairwise post-hoc tests of adjusted means have been corrected for multiple testing using Tukey’s test. Significant P values (P < 0.05) are highlighted in bold.

Analysis / Source of variation / df / MS / F / P
Chymotrypsin in the tissues
Salivary glands / Treatment / 3 / 559.658 / 1.945 / 0.126
Wet weight
P = 0.521 / Initial wet weight / 1 / 41.077 / 0.143 / 0.706
Residual / 114 / 287.809
Foregut / Treatment / 3 / 60,344.149 / 3.652 / 0.015 / pC vs Pc / 0.031
Initial wet weight
P = 0.980 / Initial wet weight / 1 / 991.241 / 0.060 / 0.807 / pC vs PC / 0.020
Residual / 114 / 16,522.503 / pC vs pc / 0.314
Pc vs PC / 1.000
Pc vs pc / 0.667
PC vs pc / 0.576
Caeca / Treatment / 3 / 200,800.431 / 5.492 / 0.001 / pC vs Pc / 0.002
Wet weight
P = 889 / Initial wet weight / 1 / 7,697.533 / 0.211 / 0.647 / pC vs PC / 0.031
Residual / 114 / 36,564.017 / pC vs pc / 0.007
Pc vs PC / 0.761
Pc vs pc / 0.968
PC vs pc / 0.951
Midgut / Treatment / 3 / 26,086.743 / 0.987 / 0.402
Wet weight
P = 0. 485 / Initial wet weight / 1 / 24,936.578 / 0.943 / 0.334
Residual / 114 / 26,442.355
Hindgut / Treatment / 3 / 4,452.101 / 1.279 / 0.285
Wet weight
P = 0. 541 / Initial wet weight / 1 / 8,025.480 / 2.305 / 0.132
Residual / 114 / 3,482.120
Amylase in the tissues
Salivary glands / Treatment / 3 / 1.594 x 108 / 13.216 / < 0.001 / pC vs Pc / < 0.001
Wet weight
P = 0.187 / Initial wet weight / 1 / 1.185 x 108 / 9.827 / 0.002 / pC vs PC / < 0.001
Residual / 104 / 12,059,088.42 / pC vs pc / 0.006
Pc vs PC / 0.712
Pc vs pc / 0.043
PC vs pc / 0.359
Amylase in the tissues
Foregut / Treatment / 3 / 73,807,857.793 / 2.121 / 0.102
Wet weight
P = 0.609 / Initial wet weight / 1 / 95,743,466.13 / 2.751 / 0.100
Residual / 103 / 34,802,235.14
Caeca / Treatment / 3 / 66,354,355.21 / 1.667 / 0.179
Wet weight
P = 0.344 / Initial wet weight / 1 / 1.779 x 108 / 4.469 / 0.037
Residual / 104 / 39,800,787.02
Midgut / Treatment / 3 / 29,943,261.738 / 1.321 / 0.272
Wet weight
P = 0.445 / Initial wet weight / 1 / 13,788,443.45 / 0.608 / 0.437
Residual / 104 / 22,665,761.37
Hindgut / Treatment / 3 / 1.270 x 108 / 7.926 / < 0.001 / pC vs Pc / 0.001
Wet weight
P = 0.548 / Initial wet weight / 1 / 5,827,224.42 / 0.364 / 0.548 / pC vs PC / 0.049
Residual / 103 / 16,025,537.55 / pC vs pc / 0.997
Pc vs PC / 0.565
Pc vs pc / 0.001
PC vs pc / 0.027
Amylase in the contents
Foregut / Treatment / 3 / 1.783 x 1010 / 5.209 / 0.002 / pC vs Pc / 0.014
Wet weight
P = 0.053 / Initial wet weight / 1 / 7.114 x 1010 / 20.778 / < 0.001 / pC vs PC / 0.002
Residual / 104 / 3.424 x 109 / pC vs pc / 0.042
Pc vs PC / 0.952
Pc vs pc / 0.965
PC vs pc / 0.735
Caeca / Treatment / 3 / 2.787 x 1010 / 8.324 / < 0.001 / pC vs Pc / < 0.001
Wet weight
P = 438 / Initial wet weight / 1 / 1.161 x 1011 / 34.670 / < 0.001 / pC vs PC / 0.072
Residual / 104 / 3.348 x 109 / pC vs pc / 0.999
Pc vs PC / 0.252
Pc vs pc / < 0.001
PC vs pc / 0.088
Midgut / Treatment / 3 / 1.235 x 1010 / 3.248 / 0.025 / pC vs Pc / 0.020
Wet weight
P = 0.188 / Initial wet weight / 1 / 1.193 x 1011 / 31.359 / < 0.001 / pC vs PC / 0.323
Residual / 104 / 3.804 x 109 / pC vs pc / 0.094
Pc vs PC / 0.561
Pc vs pc / 0.902
PC vs pc / 0.920
Hindgut / Treatment / 3 / 2.563 x 1010 / 9.484 / < 0.001 / pC vs Pc / < 0.001
Wet weight
P = 0.157 / Initial wet weight / 1 / 3.049 x 1010 / 11.284 / 0.001 / pC vs PC / 0.044
Residual / 103 / 2.702 x 109 / pC vs pc / 0.582
Pc vs PC / 0.064
Pc vs pc / 0.001
PC vs pc / 0.484

Supplementary Table 4 ANCOVA-adjusted (initial wet weight) weights of each region of the gut given dietary treatment and time. Pairwise post-hoc tests of adjusted means have been corrected for multiple testing using Tukey’s test. Significant P values (P < 0.05) are highlighted in bold.

Analysis / Source of variation / df / MS / F / P
Salivary glands / Treatment / 3 / 3.42 / 6.74 / < 0.001 / pC vs Pc / < 0.001
Time / 2 / 0.51 / 1.00 / 0.372 / pC vs PC / 0.009
Treatment * Time / 6 / 0.68 / 1.35 / 0.244 / pC vs pc / 0.022
Initial wet weight / 1 / 16.66 / 32.84 / < 0.001 / Pc vs PC / 0.653
Error / 104 / 0.51 / Pc vs pc / 0.449
PC vs pc / 0.987
Foregut / Treatment / 3 / 21.774 / 2.347 / 0.077
Time / 2 / 29.291 / 3.158 / 0.047
Treatment * Time / 6 / 3.751 / 0.404 / 0.875
Initial wet weight / 1 / 69.893 / 7.535 / 0.007
Error / 105 / 9.276
Caeca / Treatment / 3 / 7.000 / 0.385 / 0.764
Time / 2 / 25.201 / 1.386 / 0.255
Treatment * Time / 6 / 14.030 / 0.772 / 0.594
Initial wet weight / 1 / 31.460 / 1.730 / 0.191
Error / 102 / 18.180
Midgut / Treatment / 3 / 4.951 / 0.717 / 0.544
Time / 2 / 1.529 / 0.221 / 0.802
Treatment * Time / 6 / 5.969 / 0.864 / 0.524
Initial wet weight / 1 / 0.975 / 0.141 / 0.708
Error / 103 / 6.905
Hindgut / Treatment / 3 / 5.991 / 0.684 / 0.564
Time / 2 / 12.193 / 1.393 / 0.253
Treatment * Time / 6 / 1.796 / 0.205 / 0.974
Initial wet weight / 1 / 26.376 / 3.013 / 0.086
Error / 105 / 8.754