PHD-Department of Medical Laboratory Sciences
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Item Characterization of Circulating Hepatitis B Virus Genotypes among Inmates and Low Risk Voluntary Blood Donors in Garissa County, Kenya(Kenyatta University, 2023-12) Odallo, Vincent BahatiHepatitis B virus (HBV) infection is a life-threatening infection that attacks the liver and can cause both acute and chronic disease. This creates a high risk of death from cirrhosis and liver cancer. Hepatitis B infection poses a major health concern globally. It is estimated that in 2019, about 296 million people had been chronically infected with HBV globally and about 820, 000 deaths from hepatocellular carcinoma and cirrhosis were reported. In Kenya, HBV prevalence stands at chronic states of intermediate range (5-7%) and high (>8%) with regional variations. Garissa County carries a high HBV infection risk with a reported prevalence of 14.1% in pregnant women attending antenatal care (ANC) clinics. This study sought to determine the seroprevalence and circulating Hepatitis B virus genotypes among the prisoners and compare it with that of the surrounding general population at Garissa County Referral Hospital in Garissa, Kenya. A total of 130 in-mates and 130 voluntary blood donors were randomly sampled in this study and a questionnaire used to collect data on their socio-demographic characteristics. Blood was then collected in plain vacutainers and the resulting serum tested for Hepatitis B surface antigen (HBsAg) using a rapid test strip. HBV DNA was then extracted from the serum and a 930bp region of the overlapping HBV P/S gene amplified and sequenced using Sanger method. Logistic regression was used to evaluate socio-demographic factors associated with HBV seropositivity. Resulting HBV DNA sequences were manually edited, assembled into contigs and then aligned. Genotype identity of the aligned sequences was initially made using the Geno2pheno [hbv] 2.0 online data base. The aligned sequences from this study and genotype specific sequences in Genbank were then used to infer phylogenetic relationships of the local strains. The Geno2pheno [hbv] 2.0 online data base was further used to identify mutations in the samples associated with diagnostic failure and vaccine failure (in the S gene) and antiviral resistance (in the P gene). Majority of the study participants were males (86.9% among inmates and 95.4% among blood donors. Majority (76.2%) of the in-mates and of the donors (83.1%) were aged between 20-40 years while majority (51.4 % of the donors and 81.5% of in mates) had only a primary school level of education. HBV seroprevalence was significantly higher among in-mates compared to blood donors. Out of the total number of in-mates tested, 7 (5.4%) were HBYV seropositive. Conversely, among blood donors 4 (3.1 %) were seropositive. There was a significant association between HBV seropositivity and gender among both the blood donors and inmates. Out of the 22 HBV-DNA positive samples, genotype D was the most prevalent among both the in-mates and blood donors (81.8%). Genotype A sub-genotype Al was only detected among the in-mate population (18.2%). All the genotype D sequences were found to be either D/A or D/E recombinants. HBsAg escape mutations associated with diagnostic failure were noted among both study populations. As a pioneer study on HBV in Kenyan prisons, the current study confirms the categorization of in-mates as HBV high risk populations. The dominance of HBV genotype D is a novel finding as studies in other regions of the country have identified genotype A as the most prevalent. To control spread of HBV infection, serological testing should be followed up with PCR testing methods to avoid diagnostic failure.Item The phytochemical composition, efficacy and safety of herbal formulations used for management of HIV/AIDS in Mombasa County, Kenya(Kenyatta University, 2018-02) Mwavita, Amos LewaKenya has the fourth largest HIV epidemic in the world with adult prevalence of 5.9% (2016) (UNAIDS). Currently 80% of HIV patients access ARVs, while the rest do not due to stigma and traditional beliefs. In the coast of Kenya some patients seek alternative treatments to manage HIV/AIDS. Pwani herbs clinic in Mombasa is a popular clinic that dispenses herbal medicines to manage HIV and AIDS. Unfortunately, these medicines have not been subjected to formal clinical research to ascertain their efficacy and safety. This study‘s objective was to isolate the components of the herbal formulations VIRAD (Antiretroviral herbal formulation) and IMB (immune boosting herbal formulation) and to determine their efficacy on immunological parameters and safety on liver and kidney parameters in the treatment for people living with HIV and AIDS (PLWHA) in Mombasa County. Herbal formulations were analyzed for phytochemical components by qualitative and quantitative screening, mineral compositions by Atomic absorption spectrophotometer (AAS) Model: 210VGP. Patients that met the inclusion criteria and for whom written/oral informed consent was obtained and were on treatment with these herbal formulations were purposely sampled as they visited the herbal clinic. Patients were evaluated for anthropometric screening including phenotypic indices, hematological, immunological parameters CD4 /CD8 and viral load tests, liver function tests, endocrine related; were screened for a period of 12 months; including HIV disease progression, quality of life and adverse side effects. Results revealed Phytochemical components (mg/100g) Virad; Phenols 84.4mg, Saponins 531mg, Tannins 324mg, Alkaloids 2304mg, Flavonoids 2173mg, Protein 442mg and Lipids 2444.2mg. Phytochemical components (mg/100g) IMB; Phenols 75.1mg, Saponins564mg, Tannins 51mg, Alkaloids 1531mg, Flavonoids 2533mg, Proteins 544mg, Lipids 2553.8mg, characterised as metabolites antioxidant, anti-inflammatory anticholinesterase, immune boosting and antiviral agents. Trace elements in VIRAD Mineral levels (μg/g) K 12922±103Na 475±4 Ca8861±71 Mg547±1 Fe361±4 Cu 98.6±1.5 Zn43.7±0.9. Trace elements in IMB Mineral levels (μg/g) K 12085±87 Na 598±11 Ca 6047±46 Mg 545±1 Fe348±4 Cu 5.9±0.4 Zn25.3±0.6. Patients with subclinical deficiencies of trace elements would be at risk of impaired immune function. A total of 188 patients were purposely recruited into this longitudinal study. They were followed every two months for a period of one year. At months 2, 4, 6, 8 and month 10, 23/188 was lost to follow-up. Out of these three 3/23 died while twenty 20/23 developed other complications such as tuberculosis and cancer. Those who developed complications were referred for specialized treatment. Others were advised to use conventional antiretroviral drugs. At baseline were two peaks for CD4 levels 300-350 and 200-250. At completion of study weight gained by average 17kg, BMI increase 4.85, CD4 raise 126, CD8 declined 15, CD4/CD8 raise 0.19, Viral load drop 864, Hemoglobin Hb raise 3g/dl, RBC raise 1.3, WBC raise 1.6, ESR drop 21. Toxicity on kidney Urea drop 0.1g/dl, Liver GGT, AST, ALT drop0.3g/dl. Flavonoids, Alkaloids and lipids were of highest concentrations in these formulations. Immunological factors CD4/CD8 were raised; viral burdens reduced therefore improved immunity and were efficacious. Liver and kidney functions were not adversely altered; these phytochemicals were not toxic and therefore safe for use.Item Molecular Epidemiology and Evolution of Influenza A(H1N1) pdm09 Virus in Kenya(2014-02-17) Maina, George Gachara; Bulimo, Wallace D.; Otieno, Micheal FrederickIn April 2009, a novel influenza A virus was detected in Mexico and California. Thereafter, it spread globally to cause the first influenza pandemic of the 21st century. The virus was found to be antigenically unrelated to human seasonal influenza viruses but genetically related to viruses known to circulate in pigs. In view of its likely swine origin, it was initially referred to as ‘swine-origin influenza virus’ (S-OIV) A/H1N1, or pandemic influenza A (H1N1) 2009 virus but was later renamed by the World Health Organization as influenza A(H1N1)pdm09 virus. In its first year of circulation, the virus is estimated to have caused between 151,700 and 575,400 deaths globally. In Kenya, the virus was first detected in late June 2009 and in the next one year was the dominant virus in circulation. However, no laboratory confirmed deaths occurred in the country. The emergence and subsequent rapid global spread of this influenza virus provided a unique opportunity to observe the evolutionary dynamics of a pandemic influenza virus in Kenya, a tropical region where the virus circulates throughout the year. Understanding the evolution of influenza A(H1N1)pdm09 virus within the country is essential for studying global diversification, the emergence, spread and resistance of the viruses circulating in this region of the world, as well as determining the genetic relationships among the Kenyan strains and vaccine strains. The aim of this study therefore was to reconstruct the evolutionary dynamics of the A(H1N1)pdm09 influenza virus in Kenya during its first year of circulation. To accomplish this, the study undertook whole genome Sanger sequencing of 40 influenza A(H1N1)pdm09 virus isolates sampled nationwide during the pandemic period. To understand the evolutionary dynamics of the local A(H1N1)pdm09 viruses, the study employed the Bayesian evolutionary framework to analyze the resulting 320 individual gene sequences and the 40 complete genomes and compared them with sequences from two African countries, UK, USA and China isolated during the same period. The phylogenetic analyses showed that all of the Kenyan sequences sampled in the pandemic period grouped into at least four highly significant clusters and were interspersed with isolates from other countries. Two global clades (2 and 7) were identifiable within the first two weeks of the pandemic in Kenya, with clade 7 undergoing further diversification while clade 2 was not detected beyond the introductory foci. The time of the most recent common ancestor of the strains circulating in Kenya was estimated to be between April and June 2009, two months before the first laboratory confirmed case. High evolutionary rates and fast population growths was also observed. Progressive drift away from the vaccine strain was observed at both the nucleotide and amino acid level, with 2010 strains clustering separate from 2009 strains. A few unique clusters of amino acid changes were identified among all gene segments in the course of the pandemic, but no mutations previously associated with increased virulence were detected. The local strains were shown to be sensitive to neuraminidase inhibitors but resistant to adamantanes. Overall, results from this study indicate that two clades of influenza A(H1N1)pdm09 virus were introduced in Kenya and that the pandemic was sustained by multiple importations. They also indicate that clade 7 viruses dominated local transmission with an efficient community spread that was devoid of any spatial patterns but a progressive genetic drift was evident. In conclusion, adaptive evolution and viral migration seem to play a vital role in shaping the evolutionary dynamics of local A(H1N1)pdm09 viruses.