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BackgroundAedes albopictus is an important dengue vector because of its aggressive biting behavior and rapid spread out of its native home range in Southeast Asia. Pyrethroids are widely used for adult mosquito control, and resistance to pyrethroids should be carefully monitored because vector control is the only effective method currently available to prevent dengue transmission. The voltage-gated sodium channel gene is the target site of pyrethroids, and mutations in this gene cause knockdown resistance (kdr). Previous studies reported various mutations in the voltage-gated sodium channel (VGSC) gene, but the spatial distribution of kdr mutations in Ae. albopictus has not been systematically examined, and the association between kdr mutation and phenotypic resistance has not been established.MethodsA total of 597 Ae. albopictus individuals from 12 populations across Asia, Africa, America and Europe were examined for mutations in the voltage-gated sodium channel gene. Three domains for a total of 1,107 bp were sequenced for every individual. Two populations from southern China were examined for pyrethroid resistance using the World Health Organization standard tube bioassay, and the association between kdr mutations and phenotypic resistance was tested.ResultsA total of 29 synonymous mutations were found across domain II, III and IV of the VGSC gene. Non-synonymous mutations in two codons of the VGSC gene were detected in 5 populations from 4 countries. A novel mutation at 1532 codon (I1532T) was found in Rome, Italy with a frequency of 19.7%. The second novel mutation at codon 1534 (F1534S) was detected in southern China and Florida, USA with a frequency ranging from 9.5-22.6%. The WHO insecticide susceptibility bioassay found 90.1% and 96.1% mortality in the two populations from southern China, suggesting resistance and probable resistance. Positive association between kdr mutations with deltamethrin resistance was established in these two populations.ConclusionsTwo novel kdr mutations, I1532T and F1534S were found in Ae. albopictus. This is the first report of I1532T mutations in Italy and F1534S mutation in China and US. Significant association between kdr mutation and protection from deltamethrin raised the possibility that kdr mutation may be a viable biomarker for pyrethroid resistance surveillance in Ae. albopictus. The patchy distribution of kdr mutations in Ae. albopictus mosquitoes calls for developing global surveillance plan for pyrethroid resistance and developing countermeasures to mitigate the spread of resistance.

The development of Macrosiphoniella sanborni was studied at 15-35 degrees C and 80% RH, and the data obtained were used to determine the rate and threshold of its development and the sum of effective temperature. The optimal temperature for M. sanborni development ranged from 22 degrees C to 25 degrees C. After fitting the data to Wang Rusong's model, the development rate V(t) increased with rising temperature. The 2nd and 3rd instar nymphs were found more tolerable to extreme temperature and had higher optimal temperature than 1st and 4th instar nymphs. The temperature tolerance value delta was 6.70 for the 2nd and 3rd instars, and 4.70 and 4.50 for the 1st and 4th instars, respectively. The optimal temperature for the 2nd and 3rd instar was 21.76 degrees C, and the highest development temperature was 40.97 degrees C, while for the 1st and 4th instar, the optimal temperature was 23.76 degrees C and 22.49 degrees C, and the highest development temperature was 39.97 degrees C and 40.56 degrees C, respectively. Calculations with direct optimum estimation method showed that the threshold of development decreased with the aging of nymphs, while the effective accumulative temperature increased. For the 1st, 2nd, 3rd and 4th instar, the threshold of development was 6.93, 5.02, 4.58 and 4.46 degrees C, and the effective accumulative temperature was 26.88, 33.41, 33.63 and 48.49 degrees C x d(-1), respectively.