Fig 1.
The placement of the MS-300 monitors, its internal structure, and the surveillance and identification flowchart.
(A) The pink area on the map indicates the cities where monitoring points are located, while the gray area represents cities where no monitoring points have been established. The map data we used is derived from the public information of government departments (https://zhejiang.tianditu.gov.cn/standard). (B) Schematic diagram of the internal structure of the MS-300 monitor [26]. The operational mechanism is as follows: The independently developed insect lure volatilizes due to the fan-generated airflow at the bottom, attracting mosquitoes to the infrared detection window above. They are then drawn into the device through the detection window for automatic counting and fall into a storage net bag. The data is transmitted in real time to the cloud server via 4G. The icons for Culex pipiens mosquito, fever mosquito, house mosquito, tiger mosquito, and PCR cycle were provided by DBCLS (https://togotv.dbcls.jp/en/pics.html). The microtube-open-translucent icon was provided by Servier through https://smart.servier.com/. All these icons are derived from open-source resources available at https://bioicons.com.
Table 1.
Geographical coordinates of each monitoring machine.
Table 2.
Primers used in multiplex PCR and DNA barcoding.
Fig 2.
The scatter plot of mosquitoes monitored by all the MS-300 monitors from May 1st to December 31st.
Pink dots indicate mosquito appearance times, while gray dots represent sunrise and sunset times for each monitoring point (based on Beijing time). (A) Scatter plot displays the time and date of all mosquitoes detected at all the monitors. (B) Scatter plot displays the time and date of all mosquitoes monitored at each monitoring point. There were some data missing for Binjiang1233 from August 4th to August 27th.
Fig 3.
The density curves of mosquitoes monitored by all the MS-300 monitors from May 1st to December 31st.
(A) The density distribution of each monitor is showed throughout the monitoring period, on a monthly basis, for 24 hours per day. (B) The distribution of mosquito density from each monitor throughout the entire monitoring period. The pink area represents the area under the curve of mosquito vector density from May 15th to August 15th, with the displayed numbers reflecting its proportion throughout the entire monitoring period. The calculation method is as follows: Divide the total number of mosquitoes recorded at each monitoring site from May 15th to August 15th by the total number of mosquitoes recorded throughout the monitoring period (from May 1st to December 31st). There was a part of data missing for Binjiang1233 from August 4th to August 27th, which resulted in an underestimation of the calculated ratio for Binjiang1233 as shown in Fig 3B.
Fig 4.
The sensitivity and specificity of multiplex PCR.
LaneNC: RNase free water; Lane M1, GL2000 DNA marker; Lane M2, DL500 DNA marker; A: Results of multiplex PCR reactions of six mosquito species. Lane1 ~ 7, An. sinensis; Cx. p. pallens; An. anthropophagus; Ae. albopictus; Ar. subalbatus; Ae. aegypti; mixed DNA templates. B: Results of Multiplex PCR Specificity Experiment. B1 (An. sinensis Primers): Lane1, An. sinensis; Lane2 ~ 5, Cx. p. pallens; An. anthropophagus; Ae. albopictus; Ae. aegypti; B2 (Cx. p. pallens Primers): Lane1, Cx. p. pallens; Lane2 ~ 5, An. sinensis; An. anthropophagus; Ae. albopictus; Ae. aegypti; B3 (An. anthropophagus Primers): Lane1, An. anthropophagus; Lane2 ~ 5, An. sinensis; Cx. p. pallens; Ae. albopictus; Ae. aegypti; B4 (Ae. albopictus Primers): Lane1, Ae. albopictus; Lane2 ~ 5, An. sinensis; Cx. p. pallens; An. anthropophagus; Ae. aegypti; B5 (Ar. subalbatus): Lane1 ~ 5, Ar. subalbatus Primers; An. sinensis Primers; Cx. p. pallens Primers; An. anthropophagus Primers; Ae. albopictus Primers; B6 (Ae. aegypti Primers): Lane1, Ae. aegypti; Lane2 ~ 5, An. sinensis; Cx. p. pallens; An. anthropophagus; Ae. albopictus; C: Results of Multiplex PCR Sensitivity Experiment. C1 (An. sinensis), C2 (Cx. p. pallens), C3 (An. anthropophagus): Lane1 ~ 6: 1pg/µL; 100fg/µL; 10fg/µL; 1fg/µL; 0.1fg/µL; 0.01fg/µL; C4 (Ae. albopictus): Lane1 ~ 6: 10pg/µL; 1pg/µL; 100fg/µL; 10fg/µL; 1fg/µL; 0.1fg/µL; C5 (Ar. subalbatus), C6 (Ae. aegypti): Lane1 ~ 6: 100pg/µL; 10pg/µL; 1pg/µL; 100fg/µL; 10fg/µL, 1fg/µL.
Fig 5.
The phylogenetic tree of field mosquitoes.