MPH / Department of Public Health / Background: South Africa presently has a very high HIV burden. It has adopted the UNAIDS “90-90-90 targets” to curb its HIV burden. This target aims to attain sustained viral suppression in 90% of all persons receiving antiretroviral therapy. This is supported by several studies. Studies to observe if patients are achieving and sustaining viral suppression in Limpopo, South Africa, are few. Objective: To investigate the viral and immunologic responses of patients in Vhembe District to highly active antiretroviral therapy (HAART) between the 1st of January 2004 and 31st of July 2016. Methodology: This was a retrospective medical record review conducted in Vhembe District in rural Limpopo. It included the medical records of 1247 individuals from Thohoyandou Community Health Centre. Analysis was done using SPSS 24.0. To model the factors associated with virologic and immunologic responses, each independent variable was tested for association with the dependent variable (viral suppression and CD4 count increase of ≥ 50 cells/μL from baseline to 6 months). The independent variables included age, year of initiation, gender, marital status, baseline BMI, haemoglobin, clinical stage and estimated creatinine clearance. The Pearson Chi square (X2) was used for all categorical independent variables and the t-test, for all continuous independent variables, to test for association. The estimate used was a 95% confidence interval, and a p-value of < 0.05 was considered significant. Results: The study showed that 52.6% of individuals were in clinical stage I at baseline. Viral suppression (viral load < 50 copies/ml) at 6 months was 64% (n = 648), 72% (n =193) at 60 months and 94% (n = 16) at 132 months. Fifty-nine percent had consistent viral suppression for a period of at least 6 months. Consistent viral suppression (viral load < 50 copies/ml on at least one consecutive occasion without any intervening viral load > 50 copies/ml) for at least 54 months was only 14%, while 2.3% had a delay in switching from a failing regimen. The mean CD4 count at baseline was 227 cells/μL, and 538 cells/μL at 60 months. The mean CD4 cell count increase from baseline to 6 months was 190 cells/μL. The immuno-virologic discordance was 27%. Patients with higher baseline CD4 count and females were significantly (p = 0.001 and 0.031 respectively) more likely to achieve viral suppression at 6 months. Those below 45 years and females were v significantly (p = 0.011 and 0.043 respectively) more likely to achieve adequate CD4 count increase at 6 months. Conclusions: The proportion of individuals with viral suppression in the District increased from 6 months onwards, and is fairly adequate. However, sustainability of viral suppression, once attained, is low. Adequate immunologic response, however, seems high. Males and age group above 45 years appear to have poorer responses to HAART.

MPH / Department of Public Health / Background: The provision of antiretroviral treatment for people living with HIV/AIDS has encountered many challenges associated with poor adherence in South African and other countries in Africa as a whole including globally. Taking ARVs Properly has shown to reduce viral load to a level where the virus becomes undetectable and these results in an increase of CD4 count cells. These decreases chances of oppotunistic infections but it requires a proper adherence and compliance to treatment which seems to be difficult to most patients on ART. Purpose: The study investigated factors contributing to clients defaulting antiretroviral treatment. Methodology: A qualitative explorative cross-sectional study design was conducted at Matoks in Capricon District, in the months of May, June and July 2017. A purposive sampling method was used to select 19 respondents whom where willing to voluntrily participate in the study from a population of People Living With HIV/AIDS (PLWHV). An indepth face to face interview was used to collect data, guided by a central question and probing. It was then analyzed by the use of eight steps of Tesch. Results: The findings revealed that women were more defaulters than men. Shortages of antiretroviral treatment and most clients were unable to collect ART on time due to lack of transport to the clinic and the long distance from their perspective homes to the clinic. Socio economic conditions and indegenious health beliefs were some of factors identified. Recommendations: extensive health education and promotion should be intensified to reach all community members of Matoks and PLWHA in terms of HIV/AIDS care and consistent taking of treatment that clients who live far away from the clinic will be able to collect the ARV treatment nearer to their place of residents. / NRF

MPH / Department of Public Health / Human Immune Deficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS) place a significant burden on the caregivers for people living with HIV/AIDS (PLWHA). Caring for PLWHA is usually carried out by community members who are recruited from the same community as the PLWHA. These community members are trained to provide services as volunteer caregivers. The caregivers face various challenges in the process of giving care to PLWHA, often inadequately assisted by relatives, friends, neighbours, private individuals, grassroots traditional and political leaders. This results in caregivers being overwhelmed by their responsibilities, making their coping process even more difficult. The aim of this study was to explore the challenges experienced by community home-based caregivers (CHBCGs) caring for people living with HIV/AIDS at Tsianda Village, in Makhado Municipality, South Africa. This study adopted a qualitative explorative design. The population for this study were all CHBCGs caring for PLWHA. Non probability purposive sampling was used to select the community home-based caregivers working at Tsianda Community Home-based care organisation. In-depth interviews, using a semi-structured interview guide, was conducted by the researcher and a voice recorder were used to record data from participants. Data saturation was reached at the 11th participant. This is when the participants were no longer giving new information. The data collected from the study were analysed thematically. The following themes emerged from data analysis: Challenges for community home-based caregivers, perceived support for community home-based caregivers, coping strategies for community home-based caregivers. Measures to ensure trustworthiness and the code of ethics to protect the rights of the participants was applied and observed. The findings of the study revealed that community home-based caregivers experience various challenges which have a negative impact on their personal life, as well as their physical and psychological wellbeing. Community home-based care also uses different strategies to cope with these challenges. It was concluded that the community home-based caregivers are experiencing serious shortages of personal protective equipment, which makes them work with the fear of being infected with the diseases. It is recommended that the DOH should take it into consideration to provide the CHBCG’s organization with enough personal protective equipment for their safety and working without fear of being infected with deadly viruses. / NRF

PhD (Microbiology) / Department of Microbiology / Introduction Human Immunodeficiency Virus (HIV-1) continues to be a global public health concern, even though Antiretroviral drugs (ARV), especially Highly Active Antiretroviral Therapy (HAART) has significantly reduced morbidity and mortality due to AIDS globally in developed and developing countries. However, there is still a great need to explore every avenue for new therapeutic interventions due to the limitations and side effects of HAART. Potential major breakthroughs for future therapeutic development were the discoveries more than 10 years ago of the role of HIV-1 co-receptors and anti-viral activities of host restriction factors such as APOBEC3G protein, which is a member of the DNA cytosine deaminase family. Entry of HIV in to the host cell is through the attachment of the viral envelope glycoprotein to the CD4 receptor, and subsequent interaction, mainly with either CCR5 or CXCR4 co-receptors. Inhibitors, such as Maraviroc, which binds to CCR5 inhibiting entry of CCR5 utilizing viruses (R5 viruses), is currently reserved for salvage therapy in many countries including South Africa. In the course of HIV infection, CXCR4 utilizing viruses (X4 viruses) may emerge and outgrow R5 viruses, and potentially limit the effectiveness of Maraviroc. Several host cell APOBEC3 genes (A3D, A3F, A3G and A3H) have been shown to restrict HIV, and the HIV viral infectivity factor (Vif) protein serves to antagonize the action of APOBEC3 proteins, promoting viral replication. The CCR5 co-receptor and the HIV Env V3 loop have also been documented as playing roles in HIV-1 disease progression. The interplay between host and viral genes still needs widespread attention, given that disease outcomes of HIV depend on many factors, including host cell genetics. Since the discovery of these genes and their role in HIV replication, many studies have been conducted that show their association with viral polymorphism. The polymorphisms found in host cell genes can have significant effects on viral replication, transmission and fitness and can also contribute to the overall diversity in HIV-1 populations. It is hypothesized that there are significant polymorphisms in HIV-1 and cellular genes that may differ among different populations. Population-based studies have tried to establish a relationship between host factors such as APOBEC3 and CCR5 polymorphism and the rate of disease progression, but most studies have focused on Caucasian populations. In vi contrast, little information is available for the effects of variation in these genes in African populations such as South Africa, where the HIV epidemic has expanded at an alarming rate. Although several population studies have focused on African Americans, these do not give us a complete picture of the potential variation in Africans, though the studies can be a good guide on which to base additional studies. A more comprehensive analysis involving different African populations will likely provide a better understanding of the mechanisms underlying host-pathogen interactions, especially in view of the fact that African Americans are primarily infected with HIV subtype B, which is rarely seen in Africa. Methodology This study characterized the genetic variability of the APOBEC3 D, F, G and H genes as well as the HIV-1 vif, in an ethnically diverse HIV-1 infected South African cohort using Next Generation Sequencing (NGS). In addition, polymorphism in CCR5 was analyzed in conjunction with an analysis of the V3 loop sequences in HIV-1 from this cohort. Genomic DNA was extracted from peripheral blood mononuclear cells (PBMCs) of 192 HIV-1 infected drug-experienced individuals who presented for routine care at the HIV/AIDS Prevention Group Wellness Clinic (HAPG) in Bela-Bela, Donald Fraser Hope Clinic (DFHC) in Vhufhuli and in local clinics in the Vhembe district of Limpopo Province, South Africa. Next generation sequencing custom based (Tn5 tagmentation and amplicon based) protocols to prepare libraries for host and HIV-1 genes were developed and validated with commercially available library preparation kits. The Tn5 tagmentation methods were used for longer DNA fragments and the custom amplicon based methods were used mainly for the shorter DNA fragments. To determine the variability of the APOBEC3 and CCR5 host genes, gene-specific primers were designed to amplify complete 12.16 kb A3D, 13.31 kb A3F, 10.74 kb A3G, 6.8 kb A3H and 1.3 kb CCR5 genes targeting the regions containing the exons. Libraries for the resulting amplicons were prepared using Tn5 transposase tagmentation methods and sequenced on an NGS Illumina MiSeq platforms generating millions of reads with good read coverage for variant calling. Single nucleotide polymorphisms (SNPs) and indels were determined, verified in dbSNPs and compared to SNPs in other populations reported in the 1000 Genome Phase III and HapMap. A Chi-square goodness-of-fit was used to verify if whether observed genotype frequencies were in agreement with the Hardy-Weinberg Equilibrium. Haplotypes and Linkage disequilibrium were inferred to determine SNP association. vii The HIV-1 vif and env V3 loop genes were also sequenced to determine their degree of variability of these genes and to infer co-receptor usage in the South African population. Gene-specific primers were designed to amplify the 579 bp Vif region and 440 bp containing the 105 bp V3 loop. Sequencing libraries from the resulting amplicons were prepared using either the Tn5 transposase or custom-based library preparation methods and sequenced on either an Illumina MiSeq or a MiniSeq platform generating millions of reads with good read coverage for variant calling. Phylogenetic analysis was done to determine the relatedness of the sequences. Major and minor variants were determined for HIV-1 and env V3 loop quasispecies was analysed for co-receptor usage; in an effort to draw inferences for the subsequent utility of Maraviroc as salvage therapy in South Africa. Results and Discussion Next generation library preparation; Tn5 tagmentation based and custom amplicon based protocols to sequence host and HIV genes were successfully developed and used to sequence and characterize variability in host cell APOBEC3D, F, G H, CCR5 and the HIV-1 vif gene and the V3 loop region of the env gene. The HIV-1 env V3 loop sequences generated (and quasispecies analyzed) were used to infer co-receptor usage in treatment-experienced individuals; in an effort to draw inferences for the subsequent utility of Maraviroc as salvage therapy in South Africa. Quality V3 loop sequences were obtained from 72 patients, with 5 years (range: 0-16) median duration on treatment. Subtypes A1, B and C viruses were identified at frequencies of 4% (3/72), 4% (3/72) and 92% (66/72) respectively. Fifty four percent (39/72) of patients were predicted to exclusively harbor R5 viral quasispecies; and 21% (15/72) to exclusively harbor X4 viral quasispecies. Twenty five percent of patients (18/72) were predicted to harbor a dual/mixture of R5X4 quasispecies. Of these 18 patients, about 28% (5/18) were predicted to harbor the R5+X4, a mixture with a majority R5 and minority X4 viruses, while about 72% (13/18) were predicted to harbor the R5X4+ a mixture with a majority X4 and minority R5 viruses. The proportion of all patients who harboured X4 viruses either exclusively or dual/mixture was 46% (33/72). Thirty-five percent (23/66) of the patients who were of HIV-1 subtype C were predicted to harbor X4 viruses (χ2=3.58; p=0.058), and 57% of these (13/23) were predicted to harbor X4 viruses exclusively. CD4+ cell count less than 350 cell/μl was associated with the presence of X4 viruses (χ2=4.99; p=0.008). The effectiveness of Maraviroc as a component in salvage therapy may be compromised for a significant number of chronically infected patients harboring CXCR4 utilizing viii viruses in the study cohort. Although from the current study a subset of patients harboring CCR5 utilizing viruses may benefit from Maraviroc, characterizing and identifying if variation in CCR5 are located at Maraviroc binding sites was of importance to investigate. The following variants; P35P, S75S, Y89Y, A335V and Y339F and their varying frequencies were detected in the CCR5 gene. The A335V variant was detected at a higher frequency of 17.4% (29/167). The G265S variant is reported for the first time in this study at 0.6% (1/167) frequency. The SNPs detected were in strong LD (D’= 1, R2 = 0.0) with minor deviation from the Hardy-Weinberg Equilibrium. These variants were not located at the binding motif of Maraviroc. The variants A335V and Y339F were detected at a higher frequency in this study than previously reported in South Africa. Variability in APOBEC3 host cell genes was also characterized in our study cohort. The following APOBEC3 variants compared to the GRCh37 consensus sequence were detected: R97C, R248K and T316T in A3D; R48P, A78V, A108S, S118S, R143R, I87L, Q87L, V231I, E245E, S229S, Y307C and S327S in A3F; S60S, H186R, R256H, Q275E and G363R in A3G and N15Δ, G105R, K140E, K121D, E178D in A3H. Minor allele frequency variants (MAF<5%); L221L, T238I, C224Y and C320Y in A3D; I87L, P97L and S229S in A3F; R256H, A109A, F119F and L371L in A3G, which are frequent in the European population, were also detected. In addition, novel R6K, L221R and T238I variants in A3D and I117I in A3F were detected. Most of the SNPs were in strong LD with minor deviation from the Hardy-Weinberg Equilibrium. Four, six, four, and three haplotypes were identified for A3D, A3F, A3G, and A3H respectively. In general, polymorphism in A3D, 3F, 3G and 3H were higher in our South African cohort than previously reported among other African, European and Asian populations. The APOBEC3 antagonist HIV-1 vif gene was also sequenced to determine the level of diversity in a South African population and also correlated with APOBEC3 variation. Functional Vif without frameshift mutation was observed in all samples except in 4 samples. The functional domain and motifs, such as Zn binding motifs, proline-rich domain, human casein kinase, and the N and C-terminal CBF interaction site were highly conserved. APOBEC binding motifs and the nuclear localization signal were less conserved in the South African HIV-1 Vif. APOBEC3 H variation strongly correlates with Vif variation. All the Vif sequences were subtype C, except one sample, which was identified as an A1/C recombinant. The vif gene in a South African population was under purifying selection, with the dS= 0.2581 and dN= 0.0684 and the dN/dS value = 0.265. There is a high genetic diversity in the South African vif gene, which may ix influence the neutralization and restriction of APOBEC genes. Conclusions In conclusion, the protocols developed in this study can be applied to amplify and sequence any host and HIV-1 genes of interest allowing much deeper and more sensitive profiling of host gene and HIV-1 genetic diversity. Our findings show that a highly significant number of chronically HIV-1 subtype C infected patients in Maraviroc-free treatment harbor CXCR4 utilizing viruses. The data is useful in the consideration of whether to include entry antagonists such as Maraviroc in alternative forms of treatment for patients failing second line treatment regimen in the study setting. The determination of co-receptor usage prior to initiation of therapy consisting of Maraviroc is suggested. Variation in the CCR5 coding region were observed at higher frequencies compare to other studies conducted in South African populations at different locations. This data may suggest that different populations in South Africa have different SNP frequencies. All the polymorphisms identified in the study were not located at the Maraviroc binding motif, therefore the subset of patient infected by R5 viruses may benefit from this drug. We have shown that significant APOBEC3 variation exists among an ethnically diverse population of South Africa by providing extensive data for 4 different A3 genes that are known to restrict HIV infection, but have only been sparsely studied in African populations. This study provides a baseline for future studies that would functionally characterize SNPs identified in this population, in order to understand the role of novel and/or low frequency variants observed. Ex vivo and in vivo studies will increase our understanding of how these variants might have cumulatively impacted the epidemic in Northern South Africa. This study also shows that there is a high level of HIV-1 Vif diversity in the study area. This diversity may impact the expression and packaging of Vif proteins, and the infectivity of HIV. In addition, a significant correlation was observed between HIV-1 Vif variation and APOBEC3 H haplotypes. / NRF

PhD (Health Sciences) / Department of Advanced Nursing Science / The HIV/AIDS is a global epidemic which affects all people, regardless of their religion, race, age, ethnicity or geographic location. The church of Africa, which hosts the largest numbers of infected and affected people, is also challenged to be involved in the response against HIV/AIDS epidemic. Church leaders are experiencing difficulties in fulfilling their role in HIV/AIDS interventions of which they have not received training. The complexity of HIV/AIDS demands a training programme that does not simply deal with symptoms, but it must address the complexities behind and in front of the spread, and earnestly seek effective ways of controlling the spread, as well as various strategies of caring for the infected and those affected with HIV and AIDS. The purpose of this study was to develop an adapted HIV/AIDS training programme for church leaders in the Limpopo Province of South Africa. A convergent parallel mixed methods design was used; quantitative and qualitative data were collected during the same phase of the research process. The population comprised of church leaders from Christian churches in the Limpopo province. A non-probability purposive sampling was used for qualitative approach, while quota sampling was used for quantitative approach. Questionnaires were used to collect quantitative data, while in-depth interviews were used to collect qualitative data in this study. Data analysis was done separately and the two sets of results were merged into an overall interpretation of the study that informed the development of an adapted HIV/AIDS training programme. The findings of the study revealed that church leaders were not trained on how they can be involved in the response against HIV/AIDS epidemic in the Limpopo province. The current training programme did not clarify church leaders’ role towards the HIV/AIDS epidemic. The curriculum development process structure by Meyer and Van Niekerk (2008), and elements outlined by Dickoff, James and Wiedenbach (1968), were adapted to develop the training programme. A developed training programme was then validated by HIV/AIDS trainers and experts in programme development. Relevant recommendations were made to encourage churches to work effectively in addressing the HIV/AIDS epidemic in the Limpopo Province. / NRF

MSc (Microbiology) / Department of Microbiology / Background: Antiretroviral treatment (ART) has grown to be one of the most effective tool in the fight to control HIV/AIDS morbidity and mortality worldwide. However, due to the emergence of drug resistant HIV, ART efficacy can be jeopardized. Drug resistant HIV strain has a potential of becoming a major public threat, as its limit treatment options on people living with HIV. With several findings worldwide reporting drug resistant HIV to be currently being transmitted to ART-naïve persons, measures have been taken to genotype drug resistant HIV prior to treatment initiation. However, in resource limited countries such measures are not executed especially in public sectors due to the costs associated with the required assays for genotyping. Objective: The objectives of the study was to establish a deep sequencing protocol (Next Generation Sequencing-NGS) using an Illumina MiniSeq Platform and subsequently apply it to genotype HIV in chronically infected drug naïve persons for resistance mutations and viral genotypes Methods: HIV positive Individuals without any exposure to ART (Treatment-naive) were recruited. Partial pol fragment (complete protease and ~1104bp reverse transcriptase) were amplified and purified. Libraries were prepared using Nextera XT library preparation kit, fragmented, tagmented, pooled and denatured then sequenced with Illumina MiniSeq instrument. Consensus sequences were derived, aligned and phylogenetically analysed. The Stanford HIV Drug Resistance Algorithm was used to infer the presence of drug resistant mutants, at the viral minority and majority population levels. Results and discussion: An NGS protocol to generate nucleotide sequences for drug resistance inference was established. No major drug resistance mutations were detected against protease, reverse transcriptase inhibitors in the study subjects investigated. Nevertheless, V179D change was observed in one patient (8.3%). V179D has been shown to impact a low-level resistance to NNRTI. On the other hand, several secondary and unusual mutations at known drug sites were detected even at minority threshold level of <20%. Conclusion: No major drug resistance mutations was detected in the drug naïve study population. This finding suggests that there is no risk of treatment failure to the investigated subjects, however it is important to assess the potential phenotypic v | P a g e significance of the identified secondary resistance mutations in the context of HIV-1 subtype C. The established NGS protocol should be applied in subsequent HIV drug resistance studies. / NRF

PhDH / Department of Public Health / Background: Since the introduction of three-tiered systems appointments, there are a large number of missed appointments among Human immunodeficiency virus-positive clients on Antiretroviral. However, no one knows why these clients missed their scheduled times. Missing of appointments predicts poor adherence and is associated with poor clinical outcomes. Objectives: The proposed study aimed at developing a strategy for reducing the missing of appointments among adults on Antiretroviral Therapy in the Limpopo Province, South Africa. The objectives of the study are to determine patients’ behaviour, the socio-environmental and economic factors that contribute to the missing of appointments and develop strategies to enhance compliance with appointments by Human immunodeficiency virus-positive clients on Antiretroviral therapy in the Limpopo Province. Method: A qualitative research design was used to address the study objectives. Non-probability purposive sampling was used to sample health care centres in Limpopo Province, patients, Professional Nurses and Community Health Workers. Individual interview and focus group discussions strengthened the triangulation of data obtained from the participants. Creswell’s model provided details for data analysis and interpretation. Trustworthiness and Ethics: Measures to ensure data quality, such as credibility, dependability, conformability and transferability, were observed. The researcher ensured compliance with ethical standards to protect the rights of the participants. Approval for this study was obtained from the University of Venda Research Ethics Committee and the Limpopo Department of Health Research Ethics. Results: The study revealed various factors that contribute to the missing of appointments by Human immunodeficiency virus positive-clients on Antiretroviral Therapy. Specific socioeconomic, behavioural, environmental and health service-related factors appear to prevent adherence to appointments. These factors include a lack of family support and client engagement, the absence of financial means, and cultural and religious beliefs. Lack of client involvement in planning their care and poor referral of clients to community health workers were ranked high as being the most contributing factors to clients missing their appointments. Strategy development: Phase 2 of this study dealt with the development of the strategy aimed at reducing the missing of appointment by adults on Antiretroviral Therapy based on the findings of the study. The strengths, weaknesses, opportunities and threats matrix was triangulated in Political, Environmental, Social, Technological and Legal analysis to develop this approach to reduce the missing of appointments among adults on Antiretroviral therapy. A transtheoretical framework illustrated how to implement the strategy. Validation of the developed strategy ensured that the system is free of errors and checked the applicability of the strategies utilising a quantitative design. A simple random sampling approach was used to select the population to participate in this study using the questionnaire developed by the researcher. Almost all respondents agreed that the strategy would facilitate reduced missing appointments by adults on Antiretroviral therapy. Recommendations: Clients involvement and engagement throughout the process of a treatment plan is essential to identify some of the barriers that might contribute to poor adherence to appointment by clients on Antiretroviral therapy. Community health workers and nurses should be capacitated with knowledge and skills to identify the clients at risk of defaulting treatment and appointments and provide counselling that will facilitate behaviour modifications. / HWSETA

Submitted in partial fulfillment of the requirements for the Degree of PhD (Community Psychology), Faculty of Arts, University of Zululand, 2008. / HIV and AIDS present a catastrophic public health threat that is reaching crisis proportions among adolescents and young adults. There is a need to educate pre adolescent female learners before they reach puberty, as females are especially at risk of contracting HIV because of the interplay of biological, economic and cultural factors. This study explored the nature and extent of the pre-adolescent female learner's knowledge, attitudes and skills regarding HIV and AIDS. An experimental research design, namely, a no-treatment group design with pre test and posttest was applied. An HIV and AIDS prevention intervention programme was designed and implemented. The objectives were to help learners acquire knowledge, attitudes and skills to make informed decisions and practice health promoting behaviours. Life skills, sex and sexuality education and HIV and AIDS as a gendered epidemic were addressed. The science and art of health education was used to frame the intervention and ensured that the learner not only received the correct message but also incorporated it into her repertoire of behaviour. The findings of the study highlighted five critical areas that were vital within an HIV and AIDS education intervention. Firstly, pre adolescent female learners were very interested in sex and sexuality and information on physiological changes during puberty must be disseminated. The issue of morality was attached to sex and sexuality and this reinforced the biopsychosocial and cultural factors that came into play. The importance of peer education was emphasised, as parents were reticent to address these issues. It was found that basic information on all dimensions of HIV and AIDS education was still a powerful tool. It was vital to incorporate more than the ABC messages and look at the social constructs of health promotion and focus on critical thinking and an education for liberation. A manual with a set of guidelines was formulated and presented to educators for use with preadolescent female learners. This would help reduce the casualties of HIV and AIDS, as early intervention would protect a vulnerable group in society.

MSc (Microbiology) / Department of Microbiology / Background: Diarrheal diseases constitute an important problem among children but also among HIV positive patients particularly in developing countries such as South Africa. Cryptosporidium infect humans and has been shown to be an important cause of infection among different types of animals. Because of its small size, Cryptosporidium can easily go through the water purification system and can easily become a cause of an epidemic. Previous studies have shown that Cryptosporidium is an important cause of diarrhea in Limpopo Province. However, very few studies have been conducted on the genetic diversity of these organisms in the region. Therefore, the aim of this study was to detect and identify the genetic diversity of Cryptosporidium species from humans and animals in Giyani situated in the northern part of South Africa and Pretoria situated in the central part of the country. Methodology: A total of 560 samples were collected from human and animals and were all screened by microscopy using modified Ziehl-Neelsen staining technique. All the samples were tested by Enzyme-Linked Immunosorbent Assay (ELISA) using the Cryptosporidium II kits from Techlab, Virginia, USA. Positive samples from microscopy and ELISA were examined by different PCR protocols including conventional PCR for amplification of Cryptosporidium oocyst wall protein (COWP) region; Real-time PCR employing SYBR Green detection format for amplification of 18S rRNA region; Real-time PCR employing Hydrolysis probes detection format for amplification of SSU rRNA region; Real-time PCR specific for amplification of C. hominis region and C. parvum region. Positive samples from real-time PCR that gave clear bands on gel electrophoresis were sent for sequencing. The sequences were analysed using Staden package software to edit the nucleotides, Bioedit and MEGA6 software were used to align sequences and draw phylogenetic trees. The SPSS software was used for statistical analysis. xiii Results: The overall prevalence of Cryptosporidium as detected by ELISA method from the samples collected from humans was 41.2% (239/580). The prevalence was higher from the rural area 73.0% (159/218) compared to the urban area 22.1% (80/362) and the difference was statistically significant (χ2 = 145.1; p = 0.0001). Due to the limited amount of samples, only 134 ELISA-positive samples were tested using real-time PCR. Of these samples, 35.8% (48/134) tested positive. Of 48 real-time positive samples 25 were successfully sequenced and two different species (C. hominis and C. muris) were identified. Of all the sequences obtained, one (4.0%) was C. muris and 20 (80%) were C. hominis isolated from rural area, whereas 16.0% (4/25) were also C. hominis isolated from samples obtained from urban area. Cryptosporidium was not associated with diarrhea in the present study. A total of 85 samples were collected from animals (52 from cattle and 33 from goats) and of these 4 (4.7%) were positive by microscopy and ELISA. All these samples were non diarrheal. Conventional PCR also detected a similar number. Of these 4 positive samples, 1 was from a male goat, while the 3 others were obtained from female adult goats. Real-time PCR detected 56.5% (48/85) positive samples. Only 12 of the 85 animal samples were diarrheal and of these 4 were positive for Cryptosporidium. The prevalence of Cryptosporidium infection was higher 68.4% (13/19) in male animals compared to female animals 53.0% (35/66). The prevalence rates in cattle and goats were 55.8% (29/52) and 60.6% (20/33) respectively. Of 48 real-time positive samples from animals, 12 (25.0%) were successfully sequenced and two species (C. parvum and C. andersoni) were identified. Of these 6 were from cattle and the other 6 were from goats. Out of the 12 samples 10 (83%) were C. parvum while 2 (17%) were C. andersoni. Of the two C. andersoni, one was from a goat and one was from a cow. Of the 10 C. parvum, 5 were from goats and 5 were from cattle. xiv In conclusion, microscopy remains the low sensitive tool for the detection of Cryptosporidium while real time PCR appeared to be far much more sensitive by detecting more samples than all the three other methods combined. Closer to the real time PCR was ELISA that detected also more samples compared to conventional PCR and microscopy. The present study identified C. muris from humans’ samples in our area for the first time. However, C. hominis remains the dominant species that infects humans in our area. Cryptosporidium species was mostly found in samples from asymptomatic individuals. In animals, C. parvum was the most commonly isolated organism while C. andersoni was identified in our region for the first time as well and occurred in both goats and cattle. Populations in the affected areas need to be made aware of the infections so that care should be taken to avoid the spread of infection in water sources or in immunocompromised individuals.

This dissertation reports on a study undertaken to determine interventions that community libraries can engage in to improve HIV/AIDS awareness. Chapter One describes the background for the study by providing information portraying the background, research problem, aim and objectives -as well as the research methodology to be followed to ensure its successful completion. Chapter Two examines the social role and responsibility of the community library - with reference to HIV/AIDS awareness. International and national sources are used to establish what role the community libraries should play in providing resources for communal benefit. Chapter Three deals with HIV/AIDS awareness interventions currently engaged in by community libraries as well as proposed interventions. A list and description of intervention types is included. This list forms the foundation for the interview guide. Chapter Four describes the interview process. Chapter Five reports on the results of the interviews. Chapter Six provides an integrated summary of the findings from the literature review and those from the interviews in the Bojanala region. Chapter Seven concludes with a summary of recommendations and suggestions for possible further research. Essentially, the study consists of an examination of the social role and responsibility of the community library regarding HIV/AIDS awareness. An international and national literature review of the types of interventions that community libraries can embark upon to promote HIV/AIDS awareness amongst community members of all age groups. An investigation - by means of interviews with librarians/library workers in the Bojanala region - on what interventions are currently undertaken to promote HIV/AIDS awareness among the community of the Bojanala region. Recommendations regarding the types of interventions that community libraries in South Africa may provide to assist in enhancing HIV/AIDS awareness. The literature review identified the social role and responsibility of the community library; interventions currently engaged in by community libraries; and pointed the way for proposed HIV/AIDS awareness interventions that community libraries could embark on as part of their social responsibility role. It was clear from the national and international literature review that the community library - with its years of experience in information acquisition, organisation, dissemination and use - is a natural ally in supporting HIV/AIDS awareness. The aim of this investigation was to determine the social role and responsibility of community libraries in the promotion of HIV/AIDS awareness. The interventions that community library services could embark on to promote HIV/AIDS awareness. The researcher investigated the interventions that the Bojanala region could embark on to comply with the vision of the Department of Social Development (2002:9) ”that both the government and civil society as a whole develop well directed and purposeful programmes to tackle the HIV/AIDS pandemic.” This was done through face-to-face interviews with librarians/library workers responsible for managing community projects. Compared with the HIV/AIDS prevalence rate in other regions and provinces (Northern Cape and Limpopo) with similar populations, the Bojanala region was identified as a region with a high HIV prevalence rate. The recommendations made by the researcher covered nine sections pertaining to HIV/AIDS interventions, namely: Users Policies HIV/AIDS information resources Collection Development Partnerships HIV/AIDS awareness programmes Interventions Promotion Staff training The recommendations that emerge from the research could be implemented in any community library in any given country with a high HIV/AIDS prevalence rate. / Information Science / (M. Tech. (Business Administration))