Masters Degrees (Entomology)

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Effect of pyrimethamine on gametocytogenesis, exflagellation and asexual growth in southern African isolates of Plasmodium Falciparum.
(1995) Tsoka, Joyce Mahlako.; Appleton, Christopher Charles.; Freese, Janet Anne.
Pyrimethamine efficacy was investigated in vitro on the blood asexual stages, the sexual stages and exflagellation in Plasmodium falciparum. Gametocytogenesis was stimulated following the standard methods on five isolates of Plasmodium falciparum. From these five isolates, RSA 2, 3 and 5 produced gametocytes which reached maturity within seven days and the gametocytes were able to exflagellate. Isolate MW2 produced young gametocytes which disappeared within ten days. NF54 produced mature gametocytes which lasted for 24 hours only. There were no statistically significant differences between the static and the synchronization methods of gametocyte stimulation for any of the isolates. The effect of pyrimethamine was investigated by adding a known concentration of the drug (For RSA 2, MW2 and NF54, l00nmol/ℓ; RSA 3 and 5, 3000nmol/ℓ pyrimethamine) to the culture medium for seven days during gametocyte stimulation. The results of this investigation show that there was gametocytocidal activity on the isolates that were used and pyrimethamine also had a schizontocidal action on NF54 and the young gametocytes of this isolate were destroyed by the drug. At concentrations which were inhibitory to asexual parasites, the drug had a sporontocidal effect on isolate RSA 2 but not on isolate RSA 5. The pyrimethamine MIC values for asexual parasites ranged from 300nmol/ℓ to > 3000nmol/ℓ (RSA 2 and 5 were not inhibited at 3000nmol/ℓ ). These results are consistent with those found in previous studies when pyrimethamine resistance was first detected in South Africa. The chloroquine MICs indicate a good correlation with the results obtained from previous drug sensitivity tests for all the isolates examined using both the 48-hour in vitro test and isotope incorporation for growth assessment. The isobolograms constructed to determine relationship between chloroquine and pyrimethamine indicated no synergism for isolates RSA 2 and 5, but the Σ relative IC[50]s indicated a weak synergism. Both the isobolograms and the Σ relative IC[50]s for the isolates RSA 6, 9 and 14 indicated an antagonistic action between chloroquine and pyrimethamine. The results obtained from this study have important implications for malaria control in South Africa.
Epidemiology of human intestinal parasites in Qwa-Qwa, South Africa.
(1995) Mosala, Thabang Innocentia.; Appleton, Christopher Charles.
This study investigated the prevalences and intensity of intestinal parasites and aspects of their epidemiology among children in the Qwa-Qwa region of the eastern Free St~te. Faecal samples of 1180 children differing socio-economic status from nine schools at altitudes varying from 1660m to 2200m were examined quantitatively by means of the formol-ether sedimentation technique. Socio-economic, and demographic characteristics for the communities served by the schools were obtained from the literature and from a questionnaire. The study showed that, the area supports a markedly low diversity of parasite infections, and at lower intensities, than low altitude areas such as the coastal plain of KwaZulu-Natal and Eastern Cape, the Northern Province, Mpumalanga and the Western Cape. The intestinal parasite fauna affecting children in Qwa-Qwa is dominated by protozoans with only few helminths and no hookworm or bilharzia. The results indicated that factors which influence the transmission of intestinal parasites in Qwa-Qwa appear to be related primarily to social, economic and cultural aspects of the peoples' lifestyles. Climatic factors were not found important. There was a significant seasonal effect on the intensities of all parasite infection, except two protozoans, Entamoeba coli and Endolimax nana. Water source, electricity, house-type and quality of meat were found to be the important socio-economic factors that influenced parasite transmission. These relationships were investigated by fitting logistic regression and generalized linear mixed models. By documenting human parasitism (above 1700m) this study provided an endpoint to the altitudinal transect conducted in 1993 in KwaZulu-Natal by Appleton and Gouws (in press). Public health authorities and Primary Health Care personnel should find this study useful when designing and implementing nutrition and parasite control. Severe ascariasis has been reported from the study area. It will help focus PHC activities in Qwa-Qwa and in the wider context of Free State Province by demonstrating the value of proper personal and environmental hygiene in the home, thereby forming the basis for intestinal parasite control at the community level.
Malaria vector populations associated with the agricultural development at Mamfene, Northern KwaZulu-Natal, South Africa.
(1995) Obala, Andrew Ambogo.; Appleton, Christopher Charles.; Sharp, Brian Leslie.; Le Sueur, David.
The irrigation farming methods on the Makhathini Flats are thought to be responsible for recent increase in malaria cases in the Mamfene area of northern KwaZulu-Natal despite ongoing malaria control activities. Their coincidence with the period of intensive fanning is an interesting one. This study was therefore carried out to determine the relationship between larval habitats and adult mosquito population in malaria transmission using Geographic Information System (GIS). Four types of breeding sites were utilised by malaria vectors in Mamfene, viz; types 1, II, III and IV. Habitat type I was tap pools, type II was irrigation spillage in agricultural land, type III was spillage outside but adjacent to the agricultural land and type IV was depression pools located both in the Balamhlanga swamp and inland. The cumulative larval density in all habitats peaked in March 1995 (20/man-hr), with irrigation spillage (type III) recording the highest density index (33/man-hr) as compared to tap pools (type I) which recorded 32.8/man-hr while other waterbodies recorded I3 .4/man-hr (type II) and O.5/man-hr (type IV) respectively. Subsamples of both larvae and adults of An. gambiae group were identified by the Polymerase Chain Reaction (PCR) technique. Of the larvae identified, 94.1 % and 5.9% (n=289) were An. arabiensis and An. quadriannulatus respectively while in the adult component, the composition was 98.7% and 1.3% (n=303) for An. arabiensis and An. quadriannulatus respectively. This confirmed An. arabiensis as the dominant malaria vector in Mamfene area while the exophilic behaviour of An. quadriannulatus was portrayed. Of the An. gambiae group dissected for parity, 51 .5%) were found parous. This is an indication that the population was old and was able to maintain transmission locally despite ongoing vector control measures. The Global Positioning System (GPS) was used to position larval and adult mosquito sampling sites. The spatial distribution of adult mosquitoes from the breeding sites were plotted using GIS soft ware (Map Info ) and the distance between the breeding sites and study houses were measured using a utility distance tool. With the aid of GIS, the adult mosquito density in houses could be used as an indicator to locate the breeding sites in the vicinity. The importance of these findings in terms of application in cost-effective malaria control cannot be over-emphasized.
Susceptibility and resistance to insecticides among malaria vector mosquitoes in Mozambique.
(2003) Casimiro, Sonia Lina Rodrigues.; Appleton, Christopher Charles.; Sharp, Brian Leslie.
Insecticide resistance in malaria vector mosquitoes reduces the efficacy of insecticide in killing and can therefore cause a major problem for malaria vector control by insecticides. In Mozambique, pyrethroid resistance in Anopheles funestus was first detected in December 1999 in the southern corner of Maputo Province. Since then, various collections have been made at selected sites throughout the country and WHO standard susceptibility tests and biochemical assays were conducted to determine the susceptibility status and the major resistance mechanisms, in the Fl generation of field collected mosquitoes. Three malaria vector species: Anopheles funestus s.s., Anopheles gambiae s.s. and Anopheles arabiensis were identified in this study by Polymerase Chain Reaction (PCR) and their distributions plotted. The susceptibility data indicate that the Anopheles funestus s.s population in southern Mozambique is widely resistant to pyrethroid and with low levels of carbamate resistance evident at six localities. No resistance to organophosphate and DDT was observed at any study sites. Biochemical tests indicate the presence of an altered acetlylcholinesterase in all collection localities with the exception of Massinga district. Elevated esterase activity with substrate a-naphthyl acetate were detect in Boane with a probable role in organophosphate resistance. Elevated GST were detected in Boane, Moamba and Catembe. Very low levels monooxygenase titres were registered in all the localities in Mozambique, which suggest that this resistance mechanism is not operating in these areas. Pyrethroid resistance in the Anopheles gambiae complex was detected only in Anopheles arabiensis from one locality. No resistant to other groups of insecticide were observed. Altered acetlylcholinesterases were registered in all collection localities and in both species: Anopheles gambiae s.s. and Anopheles arabiensis. Elevated esterase with substrate a-naphthyl acetate were detected in Anopheles arabiensis at only one locality. Elevated GSTs were detected at all localities and in both species. The implications of the findings for malaria vector control in Mozambique are discussed.
Urinary schistosomiasis surveillance in primary health care in South Africa.
(1998) Johnson, Caron.; Appleton, Christopher Charles.
A multifaceted Schistosoma haematobium study aimed at assessing five different diagnostic techniques of surveillance was conducted. Their use in varying operational circumstances with particular reference to Primary Health Care was conducted in three areas of varying prevalence of disease namely; Mpolweni Mission (44.1%), Empangeni (30.3%) and Verulam (72.0%), KwaZulu-Natal, South Africa. This study incorporated both theoretical and applied components. The theoretical components included freshwater snail surveys, review of literature of S. haematobium diagnostic techniques and an assessment of five diagnostic techniques with particular emphasis on diagnostic performance and cost analyses. Added to these components was migration and the assessment of the prevalence of disease amongst occupants of informal settlements in and around the greater Pietermaritzburg city centre. The applied component included the initiation of a holistic S. haematobium control programme based along the World Health Organisation Guidelines. The study provided insight into several of the countries health issues relevant to both schistosomiasis and other diseases and highlighted weaknesses that may hinder the successful implementation of the current National Framework for Parasite Control. The presence of urban schistosomiasis was noted for the first time in the city of Pietermaritzburg. The present schistosomiasis distribution could be influenced by the rural-urban migration that is impacting upon major metropolitan areas. Without intermediate host snail surveys and schistosomiasis prevalence surveys amongst members of the population, the real geographic distribution of the disease will not be known. The diagnostic methods that were compared included sedimentation, filtration, three brands of chemical urinalysis strips, urine colour scales and an indirect questionnaire. Sensitivity, specificity, positive predictive, negative predictive and efficiency values were determined. Using these values, diagnostic performance ranges were established. The ranges were influenced by the cut-off values used, technique and prevalence and intensity of infection of the study area. The chemical urinalysis strips at cut-off 10erythrocyes/μl (73.7% - 93.2%) were highly sensitive whereas urine colour scales (97.1% - 99.4%) and indirect questionnaire (80.4% - 90.3%) were highly specific. The relationship between the community prevalence rates measured by all five techniques varied significantly. A cost-analysis of the techniques/sample demonstrated a wide price range (20c - R4.32). Therefore their use would be dictated more by the availability of funding than by any operational advantages each individual technique may have demonstrated. Within the public health services a need for: (1) staff training programmes, (2) core staff based within the PHC system that is dedicated to parasite control and (3) a strengthening of infrastructure was demonstrated. These may be achieved via workshops, improved communication, education courses, specific time allocation to parasite programmes i.e. parasite week, project co-ordinators and the designation of tasks.

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Browsing Masters Degrees (Entomology) by Author "Appleton, Christopher Charles."