Background
Methods
Mosquito rearing and maintenance
Fungal isolates, formulation and application
Bioassay procedures
Bioassay 1: concentration
Bioassay 2: co-infection with M. anisopliae and B. bassiana
Bioassay 3: exposure time
Bioassay 4: persistence
Statistical analysis
Results
Bioassay 1: concentration
Concentrations | MST ± SE | 1 × 107 | 1 × 108 | 1 × 109 | 2 × 109 | 2 × 1010 | 4 × 1010 |
---|---|---|---|---|---|---|---|
M. anisopliae
| |||||||
Control
| 11 ± 0.32 | χ2 = 15.12 | χ2 = 24.00 | χ2 = 50.7 | χ2 = 195.11 | χ2 = 239.68 | χ2 = 261.26 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | P < 0.001* | ||
1 × 10
7
| 10 ± 0.32 | χ2 = 1.87 | χ2 = 14.46 | χ2 = 138.37 | χ2 = 197.25 | χ2 = 219.10 | |
p = 0.17 | p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | |||
1 × 10
8
| 9 ± 0.37 | χ2 = 5.95 | χ2 = 98.86 | χ2 = 166.80 | χ2 = 188.48 | ||
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||||
1 × 10
9
| 8 ± 0.43 | χ2 = 53.99 | χ2 = 119.57 | χ2 = 104.37 | |||
p < 0.001* | p < 0.001* | p < 0.001* | |||||
2 × 10
9
| 6 ± 0.15 | χ2 = 79.62 | χ2 = 104.37 | ||||
p < 0.001* | p < 0.001* | ||||||
2 × 10
10
| 4 ± 0.12 | χ2 = 3.54 | |||||
p = 0.07 | |||||||
B. bassiana
| |||||||
Control
| 11 ± 0.32 | χ2 = 25.09 | χ2 = 47.61 | χ2 = 27.13 | χ2 = 109.37 | χ2 = 251.31 | χ2 = 251.60 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||
1 × 10
7
| 10 ± 0.47 | χ2 = 4.53 | χ2 = 0.319 | χ2 = 38.97 | χ2 = 171.18 p | χ2 = 181.16 | |
p < 0.001* | p = 0.57 | p < 0.001* | < 0.001* | p < 0.001* | |||
1 × 10
8
| 9 ± 0.23 | χ2 = 1.47 | χ = 21.40 | χ2 = 158.71 p | χ2 = 168.32 | ||
p = 0.22 | p < 0.001* | < 0.001* | p < 0.001* | ||||
1 × 10
9
| 9 ± 0.58 | χ2 = 21.90 | χ2 = 112.07 p | χ2 = 114.26 | |||
p < 0.001* | <0.001* | p < 0.001* | |||||
2 × 10
9
| 6 ± 0.17 | χ2 = 88.29 | χ2 = 130.05 | ||||
p < 0.001* | p < 0.001* | ||||||
2 × 10
10
| 4 ± 0.11 | χ2 = 3.56 | |||||
p = 0.06 |
Bioassay 2: co-infection with M. anisopliae and B. bassiana
Ratio of Ma:Bb | MST ± SE | 1:0 | 1:1 | 2:1 | 4:1 |
---|---|---|---|---|---|
Increasing ratio of
M. anisopliae
| |||||
Control
| 13 ± 0.73 | χ2 = 132.57 | χ2 = 109.63 | χ2 = 134.20 | χ2 = 121.23 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||
1:0
| 6 ± 0.12 | χ2 = 0.15 | χ2 = 0.29 | χ2 = 1.37 | |
p = 0.70 | p = 0.60 | p = 0.24 | |||
1:1
| 6 ± 0.21 | χ2 = 0.65 | χ2 = 2.59 | ||
p = 0.34 | p = 0.07 | ||||
2:1
| 6 ± 0.16 | χ2 = 0.53 | |||
p = 0.46 | |||||
Increasing ratio of
B. bassiana
| |||||
Control
| 13 ± 0.73 | χ2 = 128.01 | χ2 = 109.63 | χ2 = 133.22 | χ2 = 114.34 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||
0:1
| 5 ± 0.19 | χ2 = 1.63 | χ2 = 0.004 | χ2 = 0.29 | |
p = 0.20 | p = 0.93 | p = 0.59 | |||
1:1
| 6 ± 0.21 | χ2 = 0.29 | χ2 = 3.10 | ||
p = 0.59 | p = 0.08 | ||||
1:2
| 6 ± 0.19 | χ2 = 3.24 | |||
p = 0.06 |
Bioassay 3: exposure time
Exposure time | MST ± SE | 15 min | 30 min | 1 h | 6 h |
---|---|---|---|---|---|
M. anisopliae
| |||||
Control
| 13 ± 0.32 | χ2 = 124.1 | χ2 = 154.65 | χ2 = 149.59 | χ2 = 150.98 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||
15 min
| 6 ± 0.32 | χ2 = 36.89 | χ2 = 26.07 | χ = 28.39 | |
p < 0.001* | p < 0.001* | p < 0.001* | |||
30 min
| 5 ± 0.37 | χ2 = 1.15 | χ2 = 0.57 | ||
p = 0.28 | p = 0.44 | ||||
1 h
| 5 ± 0.43 | χ2 = 0.86 | |||
p = 0.77 | |||||
B. bassiana
| |||||
Control
| 13 ± 0.32 | χ2 = 118.5 | χ2 = 97.98 | χ2 = 79.84 | χ2 = 103.81 |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | ||
15 min
| 6 ± 0.17 | χ2 = 42.68 | χ2 = 16.28 | χ2 = 14.97 | |
p < 0.001* | p < 0.001* | p < 0.001* | |||
30 min
| 5 ± 0.11 | χ2 = 0.50, | χ2 = 0.29 | ||
p = 0.23 | p = 0.59 | ||||
1 h
| 5 ± 0.10 | χ2 = 0.06 | |||
p = 0.81 |
Bioassay 4: persistence
Comparison of: | Different concentrations within same time post-application | The same concentration between different times post-application | ||||
---|---|---|---|---|---|---|
MST ± SE
|
2 × 10
9
|
2 × 10
10
|
4 × 10
10
|
14 d
|
28
| |
1 d
| ||||||
Control | 15 ± 0.95 | χ2 = 15.15 | χ2 = 43.34 | χ2 = 55.58 | χ2 = 0.08 | χ2 = 0.01 |
p < 0.001* | p < 0.001* | p < 0.001* | p = 0.77 | p = 0.32 | ||
2 × 109 | 11 ± 0.51 | χ2 = 9.32 | χ2 = 15.44 | χ2 = 5.17 | χ2 = 4.10 | |
p < 0.001* | p < 0.001* | p < 0.001* | p < 0.001* | |||
2 × 1010 | 6 ± 0.46 | χ2 = 0.001 | χ2 = 9.80 | χ2 = 8.95 | ||
p = 0.98 | p < 0.001* | p < 0.001* | ||||
4 × 1010 | 8 ± 0.39 | χ2 = 14.74 | χ2 = 18.55 | |||
p < 0.001* | p < 0.001* | |||||
14 d
| ||||||
Control | 16 ± 1.15 | χ2 = 30.48 | χ2 = 58.07 | χ = 49.02 | χ2 = 0.43 | |
p < 0.001* | p < 0.001* | p < 0.001* | p = 0.54 | |||
2 × 109 | 12 ± 0.55 | χ2 = 6.59 | χ2 = 4.08 | χ2 = 0.06 | ||
p < 0.001* | p < 0.001* | p = 0.80 | ||||
2 × 1010 | 10 ± 0.64 | χ2 = 0.21 | χ2 = 0.49 | |||
p = 0.22 | p = 0.83 | |||||
4 × 1010 | 10 ± 0.45 | χ2 = 0.003 | ||||
p = 0.96 | ||||||
28 d
| ||||||
Control | 16 ± 1.15 | χ2 = 35.94 | χ2 = 77.49 | χ2 = 62.29 | ||
p < 0.001* | p < 0.001* | p < 0.001* | ||||
2 × 109 | 13 ± 0.33 | χ2 = 11.22 | χ2 = 5.76 | |||
p < 0.001* | p < 0.001* | |||||
2 × 1010 | 11 ± 0.49 | χ2 = 0.56 | ||||
p = 0.46 | ||||||
4 × 1010 | 11 ± 0.61 |
Comparison of: | Different concentrations within same time post-application | The same concentration between different times post-application | ||||
---|---|---|---|---|---|---|
MST ± SE
|
2 × 10
9
|
2 × 10
10
|
4 × 10
10
|
14 d
|
28
| |
1 d
| ||||||
Control | 15 ± 0.95 | χ2 = 5.73 | χ2 = 20.48 | χ2 = 16.50 | χ2 = 0.08 | χ2 = 0.01 |
p < 0.001* | p < 0.001* | p < 0.001* | p = 0.77 | p = 0.32 | ||
2 × 109 | 11 ± 0.42 | χ2 = 7.03 | χ2 = 5.01 | χ2 = 0.61 | χ2 = 4.09 | |
p < 0.001* | p < 0.001* | p = 0.44 | p < 0.001* | |||
2 × 1010 | 9 ± 0.55 | χ2 = 0.24 | χ2 = 1.74 | χ2 = 6.36 | ||
p = 0.63 | p = 0.19 | p < 0.001* | ||||
4 × 1010 | 9 ± 0.79 | χ2 = 0.09 | χ2 = 6.03 | |||
p = 0.77 | p < 0.001* | |||||
14 d
| ||||||
Control | 16 ± 1.15 | χ2 = 29.08 | χ2 = 52.46 | χ2 = 49.02 | χ2 = 0.43 | |
p < 0.001* | p < 0.001* | p < 0.001* | p = 0.54 | |||
2 × 109 | 12 ± 0.44 | χ2 = 4.23 | χ2 = 7.04 | χ2 = 5.12 | ||
p < 0.001* | p < 0.001* | p < 0.001* | ||||
2 × 1010 | 10 ± 0.67 | χ2 = 0.37 | χ2 = 1.79 | |||
p = 0.54 | p = 0.18 | |||||
4 × 1010 | 10 ± 0.45 | χ2 = 0.40 | ||||
p = 0.53 | ||||||
28 d
| ||||||
Control | 16 ± 1.15 | χ2 = 47.61 | χ2 = 34.67 | χ2 = 55.76 | ||
p < 0.001* | p < 0.001* | p < 0.001* | ||||
2 × 109 | 13 ± 0.42 | χ2 = 7.56 | χ2 = 4.92 | |||
p < 0.001* | p < 0.001* | |||||
2 × 1010 | 12 ± 0.54 | χ2 = 0.249 | ||||
p = 0.11 | ||||||
4 × 1010 | 12 ± 0.64 |