Introduction
Material and methods
Insect culture and sampling
Micro-CT scanning and analysis of tomographic data
Statistical analysis
Results
Qualitative analysis of tomographic data
Morphological landmark | Temperature (°C) | 0% | 10% | 20% | 30% | 40% | 50% | 60% | 70% | 80% | 90% | 100% |
---|---|---|---|---|---|---|---|---|---|---|---|---|
15 | 154 ADD | 179.2 ADD | 204.4 ADD | 229.6 ADD | 254.8 ADD | 280 ADD | 305.2 ADD | 330.4 ADD | 355.6 ADD | 380.8 ADD | 406 ADD | |
20 | 171 ADD | 193.8 ADD | 216.6 ADD | 239.4 ADD | 262.2 ADD | 285 ADD | 307.8 ADD | 330.6 ADD | 353.4 ADD | 376.2 ADD | 399 ADD | |
24 | 161 ADD | 184 ADD | 207 ADD | 230 ADD | 253 ADD | 276 ADD | 299 ADD | 322 ADD | 345 ADD | 368 ADD | 391 ADD | |
Larval-pupal apolysis complete | − | + | + | + | + | + | + | + | + | + | + | |
Cephalopharyngeal skeleton within the insect body | Fully within | Partially within | − | − | − | − | − | − | − | − | − | |
Gas bubble | − | + | − | − | − | − | − | − | − | − | − | |
Head, wings and legs fully everted | − | − | + | + | + | + | + | + | + | + | + | |
Pupal-adult apolysis complete | − | − | − | + | + | + | + | + | + | + | + | |
Oesophagus fully developed | − | − | − | − | + | + | + | + | + | + | + | |
Position of crop | − | − | − | Anterior part of the thorax | Posterior part of the thorax | Abdomen | Abdomen | Abdomen | Abdomen | Abdomen | Abdomen | |
Shape of the adult midgut (in sagittal view) | − | Closed sack (displaced by gas bubble) | Closed sack (occupying the central body region) | Bottle | Long-necked bottle | Tube | Tube | Tube | Tube | Tube | Tube | |
Yellow body | − | + | + | + | + | + | + | + | + | − | − | |
Rectal pouch swollen (Fig. 4d–f) | − | − | − | − | − | − | − | − | + | + | + | |
Meconium in rectal pouch | − | − | − | − | − | − | − | − | ± (96/4%) | + | + | |
Indirect flight muscles attached to adult cuticle | − | − | − | − | − | − | − | − | − | ± (22/78%) | + | |
Position of thoracic ganglion | − | − | Anterior part of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | 1/3 of the thorax | |
Medulla and lobula visible | − | − | + | + | + | + | + | + | + | + | + | |
Shape of lamina (in horizontal view) | − | − | − | Bulge | Horseshoe | Horseshoe | Horseshoe or unfolded but not fully extended (13/87%) | Unfolded but not fully extended | Unfolded and fully extended | Unfolded and fully extended | Unfolded and fully extended | |
Lamina lying under the retina | − | − | − | − | − | − | − | + | + | + | + | |
Layer of crystal cones and primary pigment cells | − | − | − | − | − | − | − | − | − | + | + |
C. vicina
L. sericata
Quantitative analysis of tomographic data
C. vicina
Developmental interval (as a percentage of the total intra-puparial period) | Relative volume of pre-helicoidal midgut (average ± STD) | Relative volume of pre-helicoidal midgut (range) | Relative volume of indirect flight muscles (average ± STD) | Relative volume of indirect flight muscles (range) | Relative volume of rectal pouch (average ± STD) | Relative volume of rectal pouch (range) |
---|---|---|---|---|---|---|
0 | 0 | 0 | 0 | 0 | 0 | 0 |
10 | 0.74 ± 0.3 | 0.57–1.32 | 0 | 0 | 0 | 0 |
20 | 3.17 ± 0.38 | 2.51–3.91 | 0.12 ± 0.09 | 0.005–0.28 | 0.001 ± 0.002 | 0–0.008 |
30 | 2.66 ± 0.29 | 2.29–3.35 | 0.27 ± 0.07 | 0.15–0.38 | 0.01 ± 0.005 | 0.005–0.02 |
40 | 2.1 ± 0.17 | 1.79–2.37 | 0.64 ± 0.12 | 0.42–0.85 | 0.02 ± 0.01 | 0.01–0.05 |
50 | 1.47 ± 0.17 | 1.21–1.86 | 0.75 ± 0.15 | 0.56–0.98 | 0.07 ± 0.03 | 0.03–0.12 |
60 | 1.14 ± 0.17 | 0.84–1.55 | 1.48 ± 0.19 | 1.23–1.77 | 0.1 ± 0.06 | 0.03–0.24 |
70 | 0.88 ± 0.18 | 0.68–1.39 | 2.12 ± 0.5 | 1.4–3.17 | 0.09 ± 0.05 | 0.04–0.21 |
80 | 0.53 ± 0.11 | 0.35–0.74 | 3.37 ± 0.18 | 3.13–3.7 | 0.64 ± 0.18 | 0.38–0.95 |
90 | 0.41 ± 0.12 | 0.2–0.72 | 4.36 ± 0.73 | 3.28–5.91 | 1.35 ± 0.31 | 0.81–1.88 |
100 | 0.34 ± 0.09 | 0.19–0.49 | 4.71 ± 0.57 | 3.98–6.07 | 1.59 ± 0.22 | 1.25–2 |
Function 1 | Function 2 | Function 3 | |
---|---|---|---|
Relative volume of indirect flight muscles | −0.523611 | 0.245734 | 0.846463 |
Relative volume of the pre-helicoidal region of the midgut | 0.664104 | 0.713722 | 0.232452 |
Relative volume of rectal pouch | −0.459161 | 0.567399 | −0.722694 |
Relative percentage | 84.56 | 13.96 | 1.48 |
10% | 20% | 30% | 40% | 50% | 60% | 70% | 80% | 90% | 100% | |
---|---|---|---|---|---|---|---|---|---|---|
Constant (k) | −4.4774 | −122.495 | −87.4043 | −56.7207 | −30.3945 | −26.354 | −28.7956 | −50.9559 | −99.506 | −122.636 |
Weight (w) of the variable | ||||||||||
Indirect flight muscles | 5.80513 | 1.98545 | 2.91656 | 5.57557 | 5.97602 | 11.3821 | 16.3072 | 23.4428 | 27.9186 | 29.5502 |
Pre-helicoidal region of the midgut | 0.241616 | 75.5737 | 63.504 | 50.1271 | 35.1466 | 27.546 | 21.4719 | 12.3071 | 8.09613 | 5.92862 |
Rectal pouch | −3.32955 | −7.8996 | −6.77473 | −6.37318 | −2.63901 | −3.8278 | −6.42504 | 17.8515 | 50.9803 | 62.1799 |
Percentage | 10% | 20% | 30% | 40% | 50% | 60% | 70% | 80% | 90% | 100% | Percentage correctly classified |
---|---|---|---|---|---|---|---|---|---|---|---|
10 |
8
| 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 100 |
20 | 0 |
11
|
4
| 0 | 0 | 0 | 0 | 0 | 0 | 0 | 73.33 |
30 | 0 |
3
|
11
|
1
| 0 | 0 | 0 | 0 | 0 | 0 | 73.33 |
40 | 0 | 0 | 0 |
15
| 0 | 0 | 0 | 0 | 0 | 0 | 100 |
50 | 0 | 0 | 0 |
1
|
14
| 0 | 0 | 0 | 0 | 0 | 93.33 |
60 | 0 | 0 | 0 | 0 | 0 |
15
| 0 | 0 | 0 | 0 | 100 |
70 | 0 | 0 | 0 | 0 | 0 |
2
|
13
| 0 | 0 | 0 | 86.66 |
80 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
15
| 0 | 0 | 100 |
90 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
12
|
3
| 80 |
100 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
6
|
9
| 60 |
Total | 8 | 14 | 15 | 17 | 14 | 17 | 13 | 15 | 18 | 12 |
86.01
|
Percentage | 10% | 20% | 30% | 40% | 50% | 60% | 70% | 80% | 90% | 100% | Percentage correctly classified |
---|---|---|---|---|---|---|---|---|---|---|---|
20 | 0 |
2
| 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 100 |
30 | 0 | 0 |
2
| 0 | 0 | 0 | 0 | 0 | 0 | 0 | 100 |
40 | 0 | 0 | 0 |
2
| 0 | 0 | 0 | 0 | 0 | 0 | 100 |
50 | 0 | 0 | 0 |
1
|
1
| 0 | 0 | 0 | 0 | 0 | 50 |
60 | 0 | 0 | 0 | 0 | 0 |
2
| 0 | 0 | 0 | 0 | 100 |
70 | 0 | 0 | 0 | 0 | 0 | 0 |
2
| 0 | 0 | 0 | 100 |
80 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2
| 0 | 0 | 100 |
90 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2
| 0 | 100 |
100 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2
| 100 |
Total | 0 | 2 | 2 | 3 | 1 | 2 | 2 | 2 | 2 | 2 |
94.44
|
L. sericata
Developmental interval (as a percentage of the total intra-puparial period) | Relative volume of pre-helicoidal midgut (average ± STD) | Relative volume of pre-helicoidal midgut (range) | Relative volume of indirect flight muscles (average ± STD) | Relative volume of indirect flight muscles (range) | Relative volume of rectal pouch (average ± STD) | Relative volume of rectal pouch (range) |
---|---|---|---|---|---|---|
20 | 2.61 ± 0.38 | 2.18–3.24 | 0.04 ± 0.01 | 0.02–0.06 | 0.002 ± 0.002 | 0–0.006 |
40 | 2.33 ± 0.33 | 2.01–2.82 | 0.35 ± 0.02 | 0.35–0.38 | 0.02 ± 0.006 | 0.02–0.03 |
60 | 1.1 ± 0.04 | 1.04–1.16 | 1.59 ± 1.09 | 1.43–1.67 | 0.08 ± 0.009 | 0.07–0.09 |
80 | 0.58 ± 0.07 | 0.5–0.71 | 2.4 ± 0.16 | 2.19–2.61 | 0.81 ± 0.09 | 0.7–0.95 |
100 | 0.18 ± 0.04 | 0.11–0.24 | 4.94 ± 0.86 | 3.66–5.85 | 1.52 ± 0.59 | 1.1–2.55 |
Percentage | 10% | 20% | 30% | 40% | 50% | 60% | 70% | 80% | 90% | 100% | Percentage correctly classified |
---|---|---|---|---|---|---|---|---|---|---|---|
20 | 0 |
1
|
3
|
1
| 0 | 0 | 0 | 0 | 0 | 0 | 20 |
40 | 0 | 0 |
2
|
3
| 0 | 0 | 0 | 0 | 0 | 0 | 60 |
60 | 0 | 0 | 0 | 0 | 0 |
5
| 0 | 0 | 0 | 0 | 100 |
80 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
5
| 0 | 0 | 100 |
100 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2
|
3
| 60 |
Total | 0 | 1 | 5 | 4 | 0 | 5 | 0 | 5 | 2 | 3 |
68
|