Evaluation of image analysis for studing mite behaviour : A thesis submitted in partial fulfilment of the requirements for the Degree of Master of Applied Science at Lincoln University

The aim of this study was to investigate the usefulness of image analysis for studying mite behaviour. Image analysis was used to convert video recordings of mites' locomotory behaviour into a series of x,y coordinates that, when joined, closely resemble the paths of mites. The coordinates were also used to calculate walking speed, direction of travel, turning frequency, turn bias and tortuosity. Two experimental arenas were developed and used to study the movement of three mite species: 1) a leaf disc arena for two-spotted spider mite, Tetranychus urticae Koch and European red mite, Panonychus ulmi (Koch); and 2), a cover-slip/tack-trap arena for Typhlodromus pyri Scheuten. Two-spotted spider mite exhibited a change in locomotory behaviour through a 48 minute period. Mites exhibited a significant decline in distance travelled, whereas the mean stationary time (per four minute interval) more than doubled, and the duration of stationary events increased steadily over the same period. A reduction in sampling frequency of mite coordinates from one per second to one every two seconds and every four seconds produced a 5% and 12% 'loss' in path length respectively. Sample period length was shown to greatly influence the results produced for some of the mean parameters calculated, however, a reduction in sample length from 3000 to 1500 coordinates was not considered to cause a major loss in information. The influence of the inherent mite movement could not be ignored and made it difficult to make decisions on the 'best' sample length to use. Some strong correlations were found between parameters used to analyse mite locomotory behaviour. In particular, arithmetic mean vector length, speed, total stationary time and total distance travelled were significantly correlated with each other. Mean angular deviation and weighted mean vector length, which both measure the degree of clustering around the mean heading angle, were strongly negatively correlated. Parameters which differentiated between 'straight' and 'tortuous' mite movement were found to be mean meander, absolute mean turn and fractal dimensions. Mean meander was thought to be the most 'powerful', while coefficient of a straight line, a commonly used parameter for measuring tortuosity, did not significantly differentiate between the two different behaviours. Frequency distributions of turns and standard deviations of the three mite species were very similar. All three species had a slight bias to turning right (clockwise) rather than to the left (counter-clockwise) and for each species certain angles occurred more often than would be expected in a 'perfect' normal distribution. A similar pattern also occurred with the frequency distribution of two-spotted spider mite heading angles, in that angles which were expected to occur more often, did not, and vice versa. The potential to use saturated salt solutions to control relative humidity on the arena was` demonstrated and indicated that relative humidity is likely to have an important influence on mite behaviour. Two-spotted spider mites appeared to move more quickly in an attempt to escape the unfavourable, extreme (10% and 95% R.H. at 25°C) moisture conditions. All three mite species displayed a characteristic edge-walking behaviour around the arenas. However, when 'edge' and 'non-edge' behaviours were compared, mean meander was the only parameter (of the parameters tested) which gave a significant difference. Behavioural responses of European red mite and T. pyri to sub-lethal (field rate) esfenvalerate were investigated and the results indicated that these mites did not seek the unsprayed halves of the arenas during the first 48 minutes. However, significant differences in most behavioural parameters to esfenvalerate residues were found with European red mite when whole arenas were compared. Image analysis is an extremely useful research tool for studying mite behaviour because of its ability to measure many parameters quickly. Careful choice of the environmental conditions, the sampling framework, and interpretation of data is essential for meaningful results.