Background
Methods
Fungal maintenance, production and storage methods and tests
Mass production
Long-term storage
Germination tests
Fungal persistence after application
Basic assay methods
Test substrates
Mosquito exposure to the substrates
Persistence assay 1 – Anopheles stephensi
Persistence assay 2 – Anopheles gambiae
Statistical analysis
Results
Spore viability during storage
Treatment | Storage temperature | Weibull parameters | Spore survival (days) | |||
---|---|---|---|---|---|---|
A | G |
R
2
| LT20 | LT50 | ||
Closed storage | 22°C | 1.53x10-10 | 5.578 | 0.99 | 60 | 73 |
26°C | 6.05x10-16 | 9.477 | 0.99 | 44 | 49 | |
32°C | 1.78x10-15 | 10.49 | 0.99 | 28 | 31 | |
Open Storage | 22°C | 5.85x10-9 | 4.203 | 0.96 | 90 | 117 |
26°C | 7.76x10-11 | 5.883 | 0.98 | 55 | 67 | |
32°C | 5.94x10-13 | 9.357 | 0.99 | 22 | 25 |
Fungal persistence following application
Assay 1 – An. stephensi
Time after exposure | Treatment | Median Lethal Time days (95% C.I.)* | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM)* |
---|---|---|---|---|
Day 1
|
Control
| 10.0 (9.05-10.95) | --------- | 12.8 (± 0.48) |
Pyrethroid
| 9.0 (8.25-9.75) | 5.9 (P = 0.015) | 12.3 (± 0.48) | |
B. bassiana
| 4.0 (3.74-4.26) | 164.1 (P < 0.001) | 5.0 (± 0.00) | |
Week 1
|
Control
| 13.0 (11.92-14.01) | --------- | Not achieved |
Pyrethroid
| 13.0 (11.0-15.0) | 0.63 (P = 0.43) | Not achieved | |
B. bassiana
| 4.0 (3.73-4.27) | 107.5 (P < 0.001) | 5.3 (± 0.25) | |
Month 1
|
Control
| 13.0 (12.23-13.77) | --------- | 12.3 (± 0.25) |
Pyrethroid
| 14.0 (13.52-14.48) | 2.05 (P = 0.15) | 12.5 (± 0.96) | |
B. bassiana
| 5.0 (4.77-5.23) | 190.2 (P < 0.001) | 5.0 (± 0.00) | |
Month 2
|
Control
| 14.0 (12.88-15.12) | --------- | Not achieved |
Pyrethroid
| 13.0 (11.20-14.80) | 0.85 (P = 0.36) | Not achieved | |
B. bassiana
| 5.0 (4.68-5.32) | 162.37 (P < 0.001) | 6.0 (± 0.00) | |
Month 3
|
Control
| 10.0 (9.07-10.93) | --------- | Not achieved |
Pyrethroid
| 9.0 (8.25-9.75) | 5.7 (P = 0.017) | Not achieved | |
B. bassiana
| 4.0 (3.74-4.26) | 168.7 (P < 0.001) | 6.5 (± 0.50) | |
Month 4
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 9.0 (8.40-9.60) | 144.5 (P < 0.001) | 12.3 (± 1.03) | |
Month 5
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 18.7 (P < 0.001) | Not achieved | |
B. bassiana
(repeat exposure)
| 11.0 (10.42-11.58) | 119.8 (P < 0.001) | 13.3 (± 0.48) |
Time after exposure | Treatment | Median Lethal Time days (95% C.I.)* | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM)* |
---|---|---|---|---|
Day 1
|
Control
| 11.0 (9.89-12.11) | --------- | Not achieved |
Pyrethroid
| 6.0 (4.77-7.23) | 58.5 (P < 0.001) | 10 (± 0.82) | |
B. bassiana
| 7.0 (6.45-7.55) | 89.1 (P < 0.001) | 11.0 (± 0.71) | |
Week 1
|
Control
| 12.0 (10.55-13.45) | Not achieved | |
Pyrethroid
| Not achieved | 4.1 (P = 0.042) | Not achieved | |
B. bassiana
| 7.0 (5.99-8.02) | 42.2 (P < 0.001) | Not achieved | |
Month 1
|
Control
| 9.0 (8.25-9.75) | --------- | 12.8 (± 0.63) |
Pyrethroid
| 9.0 (8.27-9.73) | 0.96 (P = 0.33) | 12.0 (± 0.71) | |
B. bassiana
| 9.0 (8.07-9.93) | 2.48 (P = 0.12) | 12.0 (± 0.41) | |
Month 2
|
Control
| 13.0 (12.36-13.64) | --------- | Not achieved |
Pyrethroid
| 13.0 (12.22-13.78) | 0.01 (P = 0.92) | Not achieved | |
B. bassiana
| 8.0 (7.33-8.67) | 52.63 (P < 0.001) | 12.0 (± 0.58) | |
Month 3
|
Control
| 13.0 (11.52-14.84) | --------- | Not achieved |
Pyrethroid
| 11.0 (8.72-13.28) | 1.8 (P = 0.18) | Not achieved | |
B. bassiana
| 7.0 (6.55-7.45) | 63.3 (P < 0.001) | 11.0 (± 1.29) | |
Month 4
|
Control
| Not achieved | Not achieved | |
Pyrethroid
| --------- | --------- | --------- | |
B. bassiana
| Not achieved | 16.5 (P < 0.001) | Not achieved | |
Month 5
|
Control
| Not achieved | --------- | Not achieved |
Pyrethroid
| --------- | --------- | Not achieved | |
B. bassiana
| Not achieved | 4.4 (P = 0.037) | Not achieved |
Time after exposure | Treatment | Median Lethal Time in days (95% C.I.) | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM) |
---|---|---|---|---|
Day 1
|
Control
| 10.0 (9.34-10.66) | --------- | 12.8 (± 0.48) |
Pyrethroid
| 1.0 (0.00-0.00) | 198.7 (P < 0.001) | 1.0 (± 0.00) | |
B. bassiana
| 4.0 (3.64-4.37) | 105.7 (P < 0.001) | 5.5 (± 0.87) | |
Week 1
|
Control
| 10.0 (7.38-12.63) | --------- | Not achieved |
Pyrethroid
| 1.0 (0.00-0.00) | 36.6 (P < 0.001) | 10.5 (± 2.02) | |
B. bassiana
| 4.0 (3.76-4.25) | 65.5 (P < 0.001) | 5.8 (± 0.75) | |
Month 1
|
Control
| 9.0 (7.82-10.19) | --------- | 12.3 (± 0.48) |
Pyrethroid
| 1.0 (0.00-0.00) | 107.9 (P < 0.001) | 3.0 (± 1.22) | |
B. bassiana
| 7.0 (6.05-7.59) | 3.5 (P = 0.060) | 11.5 (± 1.04) | |
Month 2
|
Control
| 11.0 (10.14-11.86) | --------- | Not achieved |
Pyrethroid
| 1.0 (0.00-0.00) | 88.19 (P < 0.001) | 5.0 (± 2.31) | |
B. bassiana
| 7.0 (6.46-7.54) | 80.4 (P < 0.001) | 9.3 (± 0.75) | |
Month 3
|
Control
| 10.0 (9.07-10.93) | --------- | Not achieved |
Pyrethroid
| 9.0 (8.25-9.75) | 5.7 (P = 0.017) | 7.5 (± 3.75) | |
B. bassiana
| 4.0 (3.74-4.26) | 168.7 (P < 0.001) | Not achieved | |
Month 4
|
Control
| Not achieved | --------- | Not achieved |
Pyrethroid
| Not achieved | 41.9 (P < 0.001) | Not achieved | |
B. bassiana
| Not achieved | 11.3 (P = 0.001) | Not achieved | |
Month 5
|
Control
| Not achieved | --------- | Not achieved |
Pyrethroid
| 2.0 (1.69-2.31) | 122.9 (P < 0.001) | Not achieved | |
B. bassiana
| Not achieved | 0.92 (P = 0.34) | Not achieved |
Assay 2 – An. gambiae
Time after exposure | Treatment | Median Lethal Time in days (95% C.I.) | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM) |
---|---|---|---|---|
Day 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 5.0 (4.81-5.19) | 209.0 (P < 0.001) | 5.8 (± 0.48) | |
Month 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 4.0 (3.85-4.15) | 185.9 (P < 0.001) | 4.5 (± 0.29) | |
Month 2
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 5.0 (4.72-5.28) | 208.1 (P < 0.001) | 6.3 (± 0.25) | |
Month 3
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 6.0 (5.71-6.29) | 219.6 (P < 0.001) | 7.0 (± 0.00) | |
Month 4
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 8.0 (7.53-8.47) | 236.9 (P < 0.001) | 9.5 (± 0.65) | |
Month 5
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 10.0 (9.48-10.52) | 155.4 (P < 0.001) | 9.5 (± 0.65) | |
Month 6
|
No assessments made
| |||
Month 7
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 11.0 (10.04-11.96) | 55.5 (P < 0.001) | Not achieved |
Time after application | Dose and Formulation | Median Lethal Time in days (95% C.I.) | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM) |
---|---|---|---|---|
Day 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| 12.0 (10.28-13.72) | 28.1 (P < 0.001) | Not achieved | |
Month 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | n.s. | Not achieved | |
Month 2
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 12.6 (P < 0.001) | Not achieved | |
Month 3
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 19.0 (P < 0.001) | Not achieved |
Time after application | Dose and Formulation | Median Lethal Time in days (95% C.I.) | Log rank statistic (Significance compared to control) | Time to 80% mortality days (± 1 SEM) |
---|---|---|---|---|
Day 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 9.1 (P < 0.003) | Not achieved | |
Month 1
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 4.9 (P = 0.027) | Not achieved | |
Month 2
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | 19.1 (P < 0.001) | Not achieved | |
Month 3
|
Control
| Not achieved | --------- | Not achieved |
B. bassiana
| Not achieved | n.s. | Not achieved |