A new technique is described here to count mosquitoes using open-source software. We wanted to develop a protocol that would estimate the total number of mosquitoes from a picture using ImageJ. Adult mosquitoes from CO2-baited traps were spread on a tray and photographed. The total number of mosquitoes in a picture was estimated using various calibrations on ImageJ, and results were compared with manual counting to identify the ideal calibration. The average trap count was 1,541, and the average difference between the manual count and the best calibration was 174.11 ± 21.59, with 93% correlation. Subsequently, contents of a trap were photographed 5 different times after they were shuffled between each picture to alter the picture pattern of adult mosquitoes. The standard error among variations stayed below 50, indicating limited variation for total count between pictures of the same trap when the pictures were processed through ImageJ. These results indicate the software could be utilized efficiently to estimate total number of mosquitoes from traps.
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