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.

The two-spotted spider mite (TSM), Tetranychus urticae Koch, is an important pest of many greenhouse and field crops worldwide. The development of resistance in TSM populations to chemical acaricides, allied with public health concerns about pesticide residues, has motivated the search for alternative control measures to suppress the pest. Hyphomycetous fungi are promising agents for mite control and the fungus Beauveria bassiana (Bb) (Balsamo) Vuillemin was investigated in this study as a biocontrol agent. The principal objectives of this study comprised: a) screening Bb strains for their pathogenicity against T. urticae; b) testing the effect of adjuvants on the efficacy of Bb; c) studying the effect of plant type on persistence of Bb and the efficacy of control of Bb against T. urticae; d) evaluating the field efficacy of Bb applications against T. urticae; e) testing the compatibility of Bb with selected fungicides; and f) assessing the synergy between Bb and soluble silicon for T. urticae control. Screening bioassays of sixty-two strains of Bb identified the two most effective strains, PPRI 7315 (R289) and PPRI 7861 (R444), that caused mortality levels of more than 80% of adult mites at 9 d post-inoculation with 2 × 108 conidia ml-1. These strains performed significantly better than the Bb commercial strain PPRI 5339, in laboratory bioassays. The two strains also attacked mite eggs, causing 53.4% and 55.5% reduction in egg hatchability at 2 × 108 conidia ml-1 respectively. However, PPRI 7861 showed relatively higher production of conidia in culture and was, therefore, selected for further trials under greenhouse and field conditions. Greenhouse evaluations of the effects of two adjuvants (Break-thru® and a paraffin oil-based emulsion) on efficacy of Bb demonstrated a higher efficacy of the biocontrol agent (BCA) when it was applied with Break-thru® or the oil solution than with water alone. Moreover, Bb conidia applied in Break-thru® solution resulted in greater control of TSM than conidia applied in the mineral oil. There was also a dose-response effect and the control of TSM by Bb increased when the concentration of conidia was increased. The control of TSM by Bb in beans (Phaseolus vulgaris L), cucumber (Cucumis sativus L.), eggplant (Solanum melongena L.), maize (Zea mays L.) and tomato (Solanum lycopersicum L.) was tested in greenhouse trials. On these crops, the persistence of conidia declined over time. The rate of decline was significantly higher on maize. However, TSM mortality was positively correlated with the amount of conidia deposited on leaves immediately after spraying, rather than their persistence over time. Higher levels of mortality of TSM due to Bb application were observed on beans, cucumber and eggplants, suggesting that the type of crop must be taken into consideration when Bb is applied as a BCA. Field efficacy of Bb against mites was evaluated in two trials on eggplants. Based on assessment of population densities of mites and leaf damage assessments; both trials showed that the strain PPRI 7861 controlled TSM in the field. Two commonly used fungicides, azoxystrobin and flutriafol, were investigated in vitro tests on culture medium and laboratory bioassays on detached bean leaves (Phaseolus vulgaris L.) for their effects on Bb. Azoxystrobin (a strobilurin) was less harmful to Bb while flutriafol was found to be inhibitory. Another important finding of this study was the substantial enhancement of Bb efficacy by soluble silicon. When Bb was combined with soluble Si, the control of TSM was better than when either of the two products was applied alone. Moreover, application of soluble Si as a plant fertilizer in hydroponic water nutrient increased accumulation of peroxidase, polyphenoloxidase and phenylalanine ammonia-lyase enzymes in leaves of plants infested with TSM. Increased activity of these defense enzymes in leaves deters feeding behaviour of mites. We suggested that feeding stress renders them susceptible to Bb infection, which would explain the synergy observed between the two agents. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

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.