Graduation date: 1995

Thesis (MSc (Botany and Zoology))--University of Stellenbosch, 2007. / Honeybee queens typically inhibit the reproductive development of workers in the colony. However, African, Apis mellifera scutellata, honeybee queens seem to have little effect on neighbouring A. m. capensis honeybee workers as is evident in the huge losses of African honeybee colonies due to the invasion by ‘social parasitic’ Cape honeybees (pseudoclones). Certain factors; such as queen and brood presence, the level of colony defence and food availability may render host colonies more vulnerable to invasion by the Cape worker honeybees. In this study host African colonies were split to determine whether a “window of opportunity” existed for Cape honeybee infiltration and thus critical to the capensis problem. Nine African colonies were infected with native and pseudoclone Cape workers over different time periods; before, during and after splitting (treatments). I measured survival rates, as well as reproductive and pheromone development of introduced workers. The effect of brood pheromones on Cape worker reproduction was also examined. Approximately 70% of all workers were removed within 72 hours, a critical period to avoid detection by Cape workers. Queen absence significantly affected the success rate of intrusion and establishment by Cape honeybee workers (GLZ; Wald χ² = 4.49, df = 1, P = 0.033). 21% of 21-day old pseudoclones survived African queenless colonies and only 6% queenright colonies. Native Cape workers showed no difference in survival rates between African queenless (12%) and queenright (11%) colonies. Looking at introduction time, considerably more pseudoclone honeybee workers survived in treatment 1 than did native Cape honeybee workers while for treatment 3 the converse was true. These data show no obvious ‘window of opportunity’ surrounding the swarming process promoting Cape honeybee infiltration and establishment of African honeybee colonies, however the period immediately prior to colony fission represents the best opportunity for invasion by pseudoclones. As for ovary and mandibular gland secretion development, all surviving pseudoclones, irrespective of A. m. scutellata queen presence, fully developed their ovaries and concomitantly produced a mandibular gland secretion dominated by 9- oxo-2-decenoic acid (9ODA). Native Cape workers showed low levels of ovary development in queenright host colonies (8-17%) but this was not true for queenless colonies, with all but one worker developing their ovaries when introduced during and after splitting. Only 40% of native Cape workers introduced before splitting developed their ovaries suggesting that queen pheromones in the three days before splitting retarded ovary development in native Cape workers. These data strengthens the suggestion that the pseudoclone honeybee workers have advanced along the queen-worker developmental continuum. Preliminary studies on brood pheromones, an important factor regulating worker reproduction, indicated that Cape workers reproduce quicker and more eggs when exposed to African brood pheromones, compared to both A. m. capensis brood pheromones and no brood pheromones. Pheromones produced by African larvae therefore do not simply inhibit Cape worker reproductive development but accelerate the commencement of egg laying by these workers. On the whole, host African colonies, especially in the absence of their queen, appear vulnerable surrounding colony fission to invasion by both Cape honeybee worker populations even though there are low survival rates. I conclude that these two Cape honeybee worker populations do differ significantly regarding their reproductive capacity and ability in becoming social parasites.

No description available.

Apolipophorin-III is a hemolymph protein known for its role in lipid transport. Apolipophorin-III isolated from the hemolymph of last instar larvae of Galleria mellonella bound to the surface of the insect pathogenic Gram-negative bacterium Xenorhabdus nematophilus and to the lipid A moiety of its lipopolysaccharide. This binding reduced the toxicity of the lipopolysaccharide to hemocytes and decreased the inhibitory effect of the lipopolysaccharide on phenoloxidase. Apolipophorin-III also bound to the Gram-positive bacterium Micrococcus lysodeikticus; this enhanced the activity of hen egg lysozyme on the organism as well as the lytic activity of G. mellonella cell-free hemolymph. / The involvement of apolipophorin-III in the immune responses of G. mellonella larvae to lipoteichoic acids, surface components of Gram-positive bacteria, was examined. Lipoteichoic acids from Bacillus subtilis, Enterococcus hirae and Streptococcus pyogenes caused a dose- and time-dependent drop in the total counts of circulating hemocytes and a partial or complete depletion of plasmatocytes depending on the species of lipoteichoic acid. All lipoteichoic acids tested activated phenoloxidase in vitro; however, in vivo, only B. subtilis lipoteichoic acid elevated the phenoloxidase activity while the other two suppressed it. Binding of apolipophorin-III to lipoteichoic acids was demonstrated. Apolipophorin-III prevented the complete depletion of plasmatocytes and depressed the activation of phenoloxidase by lipoteichoic acid from B. subtilis. The concentration of apolipophorin-III in hemolymph two hours post injections of lipopolysaccharides or lipoteichoic acids into larvae of G. mellonella did not change with respect to control insects that received phosphate-buffered saline. The concentration of apolipophorin-III in hemolymph at the end of the feeding larval stage was 8--12 mg/mL of hemolymph. Apolipophorin-III was present in significant amounts in the prepupal, pupal and adult stages. The protein was detected immunologically in hemocyte lysates, plasma and fat body. Non-denaturing polyacrylamide gels and immunoblots of fresh hemolymph suggested that apolipophorin-III is associated with a 77 kDa protein.

The occurrence of Nosema disease, tracheal mites and the “pseudo-parasitic” behaviour of Cape honeybee workers when placed amongst African honeybees – known as the Cape Bee Problem – were studied over a 18 month period. Three surveys, approximately 6 months apart were done. The aims of this study were to establish the distribution and severity of the diseases and compare the disease with the presence of the Cape Bee Problem. Before this survey commenced European Foul Brood disease, Sacbrood (virus), Nosema, Brood nosema, and Tracheal mite have sporadically been reported in the summer rainfall region of South Africa. In the first survey 1005 colonies in 61 apiaries were surveyed, 803 colonies in 57 apiaries in the second, and 458 colonies in 41 apiaries in the third. Samples for disease and parasite analysis were taken at 4 colonies per apiary. Ten colonies per apiary were inspected for Cape Problem Bees, and samples of workers were collected and dissected at each of these colonies. Even with the addition of apiaries to 'fill-up' lost colonies during the second survey, 63% of all colonies were lost by the third survey. There was only a small difference in colony loss between sedentary and migratory beekeepers of 22% compared to 27%. Nosema was more prevalent amongst commercial beekeepers and increased in migratory operations during the survey period. The percentage of colonies infected increased during the survey period from 23% to 32% to 34%. The placement of colonies in Eucalyptus plantations may boost infection. Trachea mites seem to have spread quite rapidly in South Africa since its discovery. This parasitic mite was present in all regions, although in low numbers in three most northern regions. Sedentary colonies had higher levels of infestation than migratory colonies. The number of colonies infested diminished over the survey period, which may be a result of general colony loss. The Cape Problem Bee was less of a problem than anticipated. Colonies succumbed to Cape Problem Bees in all regions. When beekeepers reported high levels of infestation in their bee stocks the colonies would be dead within six months. In apiaries with low infestation the die-out was slower.