Background
Methods
Study area and mosquito sampling
DNA extraction and species identification
Protocol of identification | Years | Months | Location of capture |
An. coluzzii
|
An. gambiae
| Hybrids |
An. arabiensis
| Total |
---|---|---|---|---|---|---|---|---|
A (n = 611) | 2006 | September | Indoor | 21 | 14 | 1 | 7 | 43 |
Outdoor | 44 | 4 | 0 | 17 | 65 | |||
October | Indoor | 10 | 6 | 3 | 6 | 25 | ||
Outdoor | 18 | 5 | 0 | 9 | 32 | |||
Total | 93 | 29 | 4 (3.2%) | 39 | 165 | |||
2008 | July | Indoor | 0 | 2 | 0 | 53 | 55 | |
Outdoor | 1 | 0 | 0 | 82 | 83 | |||
October | Indoor | 14 | 3 | 1 | 35 | 53 | ||
Outdoor | 10 | 2 | 0 | 24 | 36 | |||
Total | 25 | 7 | 1 (3.03%) | 194 | 227 | |||
2010 | September | Indoor | 8 | 20 | 1 | 42 | 71 | |
Outdoor | 7 | 13 | 0 | 31 | 51 | |||
October | Indoor | 1 | 16 | 0 | 19 | 36 | ||
Outdoor | 2 | 16 | 0 | 43 | 61 | |||
Total | 18 | 65 | 1 (1.2%) | 135 | 219 | |||
Total | 136 | 101 | 6 | 368 | 611 | |||
B (n = 497) |
An coluzzii
| 109 | 2 | 0 | 1 | 112 (97.32) | ||
An. gambiae
| 4 | 86 | 0 | 0 | 90 (95.55) | |||
An. coluzzii/An. gambiae hybrids | 5 | 0 | 1 | 0 | 6 (16.66) | |||
An. arabiensis
| 0 | 1 | 0 | 288 | 289 (99.65) |
Sample species | Months/year | Location of capture | Sample codes | Sample sizes |
---|---|---|---|---|
An. coluzzii
| 2006 | Indoor | ACIND06 | 22 |
Outdoor | ACOUT06 | 57 | ||
2008 | Indoor and outdoor | AC08 | 25 | |
2010 | Indoor and outdoor | AC10 | 15 | |
An. gambiae
| 2006 | Indoor and outdoor | AG06 | 23 |
2010 | Indoor | AGIND10 | 32 | |
Outdoor | AGOUT10 | 27 | ||
An. arabiensis
| Sept 2006 | Outdoor | AROUTSEP06 | 15 |
2008 | ||||
July | Indoor | ARINDJUL08 | 44 | |
Outdoor | AROUTJUL08 | 72 | ||
Sept | Indoor | ARINDSEP08 | 33 | |
Outdoor | AROUTSEP08 | 22 | ||
2010 | ||||
Sept | Indoor | ARINDSEP10 | 38 | |
Outdoor | AROUTSEP10 | 27 | ||
Oct | Indoor | ARINDOCT10 | 19 | |
Outdoor | AROUTOCT10 | 39 |
Microsatellite genotyping
Marker locus | RP | Sequence primer flanking | AS (bp) | Ta (°C) |
An. coluzzii (N = 119) |
An. gambiae (N = 82) |
An. arabiensis (N = 309) | |||
---|---|---|---|---|---|---|---|---|---|---|
Na | A | Na | A | Na | A | |||||
H59 | (GT)9 | 5′ CCCCTATTAAACCCTGGACG 3′ 3′ TGTTGTTGCCCTGCGTTACC 5′ | 123 | 55 | 10 | 6.75 ± 1.71 | 7 | 6.00 ± 1.00 | 13 | 7.33 ± 1.50 |
H88 | (GT)9 | 5′TGCGGCGGTAAAGCATCAAC 3′ 3′ CCGGTAACACTGCGCCGAC 5′ | 176 | 60 | NPA | NPA | NPA | NPA | 23 | 13.44 ± 3.05 |
H93 | (GT)4 + 7 | 5′ TCCCCAGCTCACCCTTCAAG 3′ 3′ GGTTGCATGTTTGGATAGCG 5′ | 209 | 55 | 19 | 10.75 ± 3.86 | 20 | 12.67 ± 0.58 | 30 | 13.00 ± 3.81 |
H119 | (GT)6 | 5′ GGTTGATGCTGAAGAGTGGG 3′ 3′ ATGCCAGCGGATACGATTCG 5′ | 174 | 55 | 17 | 12.25 ± 1.26 | 14 | 10.67 ± 0.58 | 10 | 6.89 ± 1.69 |
H128 | (GT)21 | 5′ CGGGACGGCTAGATAAAGCG 3′ 3′ CCGGGCGACATAACCCACCC 5′ | 117 | 55 | 25 | 17.5 ± 3.42 | 24 | 16.67 ± 3.22 |
b
|
b
|
H242 | (GT)8 | 5′TTCATTTCCACCGCAGCTGC 3′ 3′GGCGACACTCAATCCTTCC 5′ | 69 | 58 | 7 | 5.50 ± 1.00 | 7 | 5.67 ± 0.58 |
c
|
c
|
H249 | (GT)15 | 5′ ATGTTCCGCACTTCCGACAC 3′ 3′ GCGAGCTACAACAATGGAGC 5′ | 129 | 60 | 22 | 13.50 ± 1.73 | 17 | 12.33 ± 1.16 | 27 | 12.67 ± 2.69 |
H312 | (GT)10 | 5′ TAAACCATCAACCAGCCACC 3′ 3′ ACTGTGTGCGAGTCGGTTGG 5′ | 108 | 58 | 19 | 12.75 ± 1.50 | 16 | 13.00 ± 0.00 | 29 | 15.33 ± 4.79 |
H555 | (GT)8 | 5′GCAGAGACACTTTCCGAAAC3′ 3′ TGTCAACCCACATTTTGCGC 5′ | 81 | 55 | 17 | 10.50 ± 3.00 | 15 | 9.67 ± 3.06 |
b
|
b
|
H577 | (GT)16 | 5′ TTCAGCTTCAGGTTGGTCTC 3′ 3′ GGGTTTTTTGGCTGCGACTG 5′ | 113 | 55 | 13 | 8.50 ± 2.65 | 14 | 10.00 ± 1.00 | 22 | 10.78 ± 3.31 |
H746 | (GT)14 | 5′ TGGGTTCGAAATTCGCCAAC 3′ 3′ GACGTGTGCACCCGTTGTG 5′ | 105 | 58 | 14 | 11.25 ± 1.26 | 13 | 10.67 ± 1.16 | 29 | 16.89 ± 2.57 |
H758 | (GT)11 | 5′ TGATTTGCCAGTTCTGCCAG 3′ 3′ GTGATTGGAGTGGCTAGTGG 5′ | 105 | 60 |
a
|
a
|
a
|
a
| 11 | 5.11 ± 0.93 |
H812 | (GT)10 | 5′CTGGCCCATTTTGCATATGC 3′ 3′ TGCTCCACCAAACCACATC 5′ | 131 | 55 |
a
|
a
|
a
|
a
| 15 | 10.00 ± 1.58 |
H817 | (GT)8 | 5′ ACTGGTCCGTTGCTGCGCG 3′ 3′ ATGAGTGAATGGTGCGCTGG 5′ | 124 | 58 | 7 | 6.00 ± 0.82 | 6 | 5.00 ± 1.00 |
b
|
b
|
Mean | 15.46 ± 5.87 | 10.48 ± 3.89 | 13.91 ± 5.59 | 10.21 ± 3.76 | 20.90 ± 7.56 | 11.14 ± 4.14 |
Data analysis
Results
Species composition
Genetic structure of Anopheles populations
Populations | N | A ± se | Ho ± se | He ± se | P value ± se | Fis CI 95% | IAM | SMM |
---|---|---|---|---|---|---|---|---|
ACIND06 | 22 | 9.455 ± 3.934 | 0.706 ± 0.202 | 0.785 ± 0.135 | 0.000 ± 0.000 | 0.102 (−0.004 to 0.160) | ps = 0.305 (8/3)
p
w
= 0.027 | ps = 0.036 (3/8) pw = 0.966 |
ACOUT06 | 57 | 12.455 ± 4.458 | 0.742 ± 0.113 | 0.800 ± 0.108 | 0.000 ± 0.000 | 0.073 (0.023 to 0.109) |
p
s
= 0.033 (10/1)
p
w
= 0.006 | ps = 0.00005 (0/11) pw = 1.00000 |
AC08 | 25 | 10.909 ± 3.646 | 0.734 ± 0.105 | 0.824 ± 0.107 | 0.000 ± 0.000 | 0.112 (0.042 to 0.137) | ps = 0.127 (9/2)
p
w
= 0.008 | ps = 0.033 (3/8) pw = 0.949 |
AC10 | 15 | 8.000 ± 2.898 | 0.739 ± 0.150 | 0.810 ± 0.140 | 0.000 ± 0.000 | 0.090 (−0.017 to 0.106) | ps = 0.130 (9/2)
p
w
= 0.034 | ps = 0.116 (4/7) pw = 0.861 |
AG06 | 23 | 10.091 ± 4.110 | 0.725 ± 0.164 | 0.809 ± 0.095 | 0.000 ± 0.000 | 0.106 (0.037 to 0.126) | ps = 0.118 (9/2)
p
w
= 0.034 | ps = 0.110 (4/7) pw = 0.949 |
AGIND10 | 32 | 10.182 ± 3.125 | 0.693 ± 0.136 | 0.796 ± 0.116 | 0.000 ± 0.000 | 0.131 (0.061 to 0.175) | ps = 0.130 (9/2)
p
w
= 0.034 | ps = 0.007 (2/9) pw = 0.995 |
AGOUT10 | 27 | 10.364 ± 4.056 | 0.690 ± 0.156 | 0.806 ± 0.103 | 0.000 ± 0.000 | 0.146 (0.053 to 0.203) |
p
s
= 0.031 (10/1)
p
w
= 0.027 | ps = 0.031 (3/8) pw = 0.938 |
AROUTSEP06 | 15 | 8.500 ± 3.240 | 0.560 ± 0.130 | 0.760 ± 0.137 | 0.000 ± 0.000 | 0.270 (0.134 to 0.326) | ps = 0.361 (5/5) pw = 0.500 | ps = 0.180 (4/6) pw = 0.920 |
ARINDJUL08 | 44 | 14.000 ± 5.603 | 0.738 ± 0.125 | 0.805 ± 0.086 | 0.000 ± 0.000 | 0.085 (0.021 to 0.127) | ps = 0.613 (6/4) pw = 0.500 | ps = 0.0001 (0/10) pw = 1.000 |
AROUTJUL08 | 72 | 14.400 ± 5.696 | 0.649 ± 0.097 | 0.749 ± 0.103 | 0.000 ± 0.000 | 0.135 (0.090 to 0.165) | ps = 0.354 (5/5) pw = 0.754 | ps = 0.00012 (0/10) pw = 1.000 |
ARINDSEP08 | 33 | 11.800 ± 4.237 | 0.680 ± 0.091 | 0.780 ± 0.113 | 0.000 ± 0.000 | 0.131 (0.049 to 0.179) | ps = 0.631 (6/4) pw = 0.385 | ps = 0.00014 (0/10) pw = 1.000 |
AROUTSEP08 | 22 | 8.500 ± 3.308 | 0.643 ± 0.092 | 0.749 ± 0.133 | 0.005 ± 0.002 | 0.145 (0.045 to 0.188) | ps = 0.366 (7/3) pw = 0.216 | ps = 0.171 (4/6) pw = 0.984 |
ARINDSEP10 | 38 | 11.400 ± 4.088 | 0.619 ± 0.101 | 0.755 ± 0.114 | 0.000 ± 0.000 | 0.181 (0.121 to 0.214) | ps = 0.362 (5/5) pw = 0.423 | ps = 0.00015 (0/10) pw = 1.000 |
AROUTSEP10 | 27 | 10.500 ± 3.923 | 0.692 ± 0.116 | 0.766 ± 0.117 | 0.000 ± 0.000 | 0.099 (0.020 to 0.138) | ps = 0.358 (5/5) pw = 0.461 | ps = 0.002 (1/9) pw = 0.999 |
ARINDOCT10 | 19 | 9.400 ± 3.134 | 0.730 ± 0.112 | 0.756 ± 0.098 | 0.009 ± 0.003 | 0.035 (−0.065 to 0.081) | ps = 0.366 (5/5) pw = 0.839 | ps = 0.002 (1/9) pw = 0.999 |
AROUTOCT10 | 39 | 11.700 ± 4.029 | 0.686 ± 0.06 | 0766 ± 0.104 | 0.000 ± 0.000 | 0.106 (0.046 to 0.135) | ps = 0.609 (6/4) pw = 0.423 | ps = 0.00016 (0/10) pw = 0.991 |
Genetic differentiation between populations
Populations | ACIND06 | ACOUT06 | AC08 | AC10 | AGIND06 | AGIND10 | AGOUT10 |
---|---|---|---|---|---|---|---|
ACIND06 | 0.000 | 0.449 ± 0.005 | 0.426 ± 0.006 | 0.386 ± 0.005 | 0.150 ± 0.004 | 0.081 ± 0.003 |
0.007 ± 0.001
|
ACOUT06 | 0.001 | 0.000 | 0.155 ± 0.004 | 0.161 ± 0.004 |
0.027 ± 0.002
|
0.0004 ± 0.0002
|
0.000 ± 0.000
|
AC08 | 0.003 | 0.004 | 0.000 | 0.308 ± 0.005 | 0.082 ± 0.003 | 0.091 ± 0.003 |
0.007 ± 0.001
|
AC10 | 0.004 | 0.006 | 0.005 | 0.000 |
0.013 ± 0.001
|
0.000 ± 0.000
|
0.003 ± 0.001
|
AGIND06 | 0.007 |
0.008
| 0.008 |
0.016
| 0.000 | 0.525 ± 0.006 | 0.436 ± 0.001 |
AGIND10 | 0.008 |
0.011
| 0.007 |
0.025
| 0.001 | 0.000 |
0.05 ± 0.002
|
AGOUT10 |
0.016
|
0.015
|
0.014
|
0.021
| 0.003 |
0.008
| 0.000 |
Populations | AROUTSEP06 | ARINDJUL08 | AROUTJUL08 | ARINDSEP08 | AROUTSEP08 | ARINDSEP10 | AROUTSEP10 | ARINTOCT10 | AROUTOCT10 |
---|---|---|---|---|---|---|---|---|---|
AROUTSEP06 | 0.000 | 0.273 ± 0.004 | 0.267 ± 0.005 | 0.745 ± 0.004 | 0.794 ± 0.004 | 0.777 ± 0.004 | 0.825 ± 0.004 | 0.634 ± 0.005 | 0.245 ± 0.004 |
ARINDJUL08 | 0.005 | 0.000 |
0.008 ± 0.001
|
0.030 ± 0.002
| 0.197 ± 0.005 |
0.0002 ± 0.0001
| 0.141 ± 0.003 | 0.295 ± 0.005 | 0.531 ± 0.006 |
AROUTJUL08 | 0.006 |
0.007
| 0.000 | 0.053 ± 0.002 | 0.480 ± 0.005 | 0.088 ± 0.003 | 0.846 ± 0.003 | 0.509 ± 0.006 |
0.035 ± 0.002
|
ARINDSEP08 | 0.0002 |
0.007
| 0.006 | 0.000 | 0.883 ± 0.004 |
0.015 ± 0.001
| 0.325 ± 0.005 | 0.379 ± 0.005 |
0.031 ± 0.002
|
AROUTSEP08 | 0 | 0.005 | 0.001 | −0.003 | 0.000 | 0.380 ± 0.005 | 0.808 ± 0.004 | 0.791 ± 0.004 | 0.228 ± 0.004 |
ARINDSEP10 | 0 |
0.014
| 0.005 |
0.011
| 0.004 | 0.00000 | 0.949 ± 0.002 | 0.617 ± 0.005 |
0.004 ± 0.001
|
AROUTSEP10 | 0 | 0.005 | 0 | 0.003 | 0 | 0 | 0.000 | 0.856 ± 0.004 | 0.466 ± 0.005 |
ARINDOCT10 | 0.002 | 0.003 | 0.001 | 0.003 | 0 | 0.001 | 0 | 0.000 | 0.325 ± 0.005 |
AROUTOCT10 | 0.007 | 0.00053 |
0.006
|
0.008
| 0.004 |
0.012
| 0.001 | 0.003 | 0.000 |
Assignment test
ACIND06 | ACOUT06 | AC08 | AC10 | AG06 | AGIND10 | AGOUT10 | |
---|---|---|---|---|---|---|---|
ACIND06 | 9.09 | 21.05 | 16 | 20 | 17.39 | 15.63 | 0 |
ACOUT06 | 22.73 | 36.84 | 24 | 20 | 13.04 | 6.25 | 0 |
AC08 | 18.18 | 7.02 | 8 | 26.67 | 0 | 6.25 | 7.40 |
AC10 | 13.64 | 10.53 | 20 | 13.33 | 4.35 | 0 | 7.40 |
AG06 | 18.18 | 14.04 | 12 | 6.67 | 21.74 | 9.38 | 18.52 |
AGIND10 | 13.64 | 10.53 | 12 | 0 | 26.09 | 40.63 | 22.22 |
AGOUT10 | 4.55 | 0 | 8 | 13.33 | 17.39 | 21.88 | 44.44 |
Total | 22 | 57 | 25 | 15 | 23 | 32 | 27 |
AROUT SEP06 | ARIND JUL08 | AROUT JUL08 | ARIND SEP08 | AROUT SEP08 | ARIND SEP10 | AROUT SEP10 | ARIND OCT10 | AROUT OCT10 | |
---|---|---|---|---|---|---|---|---|---|
AROUTSEP06 | 13.33 | 13.64 | 5.56 | 9.09 | 13.64 | 7.89 | 11.11 | 10.53 | 0 |
ARINDJUL08 | 13.33 | 20.45 | 13.89 | 6.06 | 0 | 2.63 | 7.40 | 0 | 10.26 |
AROUTJUL08 | 6.67 | 9.09 | 22.22 | 18.18 | 13.64 | 15.79 | 3.70 | 10.53 | 2.56 |
ARINDSEP08 | 13.33 | 4.55 | 12.5 | 18.18 | 27.27 | 2.63 | 0 | 5.26 | 7.69 |
AROUTSEP08 | 6.67 | 15.91 | 11.11 | 15.15 | 13.64 | 7.89 | 22.22 | 26.32 | 15.39 |
ARINDSEP10 | 20 | 4.55 | 15.28 | 9.09 | 9.09 | 21.05 | 22.22 | 10.53 | 12.82 |
AROUTSEP10 | 13.33 | 4.55 | 9.72 | 6.06 | 18.18 | 13.16 | 7.40 | 10.53 | 12.82 |
ARINDOCT10 | 6.67 | 6.82 | 11.11 | 6.06 | 4.55 | 15.79 | 14.81 | 10.53 | 23.07 |
AROUTOCT10 | 6.67 | 20.45 | 11.11 | 12.12 | 0 | 13.16 | 11.11 | 15.79 | 15.38 |
Total | 15 | 44 | 72 | 33 | 22 | 38 | 27 | 19 | 39 |
An. coluzzii
|
An. gambiae
| |
---|---|---|
An. coluzzii
| 75.63 (90) | 26.83 (22) |
An. gambiae
| 24.37 (29) | 73.17 (60) |
Hybrids | 66.67 (4) | 33.33 (2) |
Total | 123 | 84 |
Hybrid individuals |
An. coluzzii
|
An. gambiae
|
HYBINDOCT06 | 99.795 | 0.205 |
HYBINDOCT06 | 0.118 | 99.882 |
HYBINDOCT06 | 100.000 | 0.000 |
HYBINDSEP06 | 96.176 | 3.824 |
HYBINDSEP08 | 99.919 | 0.081 |
HYBINDSEP10 | 6.292 | 93.708 |