MST-Department of Medical Laboratory Sciences

Permanent URI for this collection

https://repository.ku.ac.ke/handle/123456789/9107

Browse

Now showing 1 - 20 of 53

Dengue 2 infection in wild caught papio anubis (olive Baboons) and chlorocebus aethiops (african green Monkey) from selected regions of Kenya
(Kenyatta University, 2014) Ambala, Peris Auma
Dengue viruses, family Flaviviridae and genus Flavivirus occur as four antigenically distinct serotypes denoted as dengue virus 1, 2, 3, 4 (DENY-I, 2, 3, 4). The four serotypes are known to broadly infect different vertebrate hosts including humans, wild animals and various species of non-human primates (NHPs). Despite the fact that NHPs have increasingly been implicated as potential sources of emerging zoonotic viral diseases, there is no available scientific data on sylvatic dengue virus surveillance in Kenya. The present study investigated the sero-prevalence of DENY-2 exposure and characterized DENY serotypes circulating in NHPs from Laikipia, Aberdares, Kajiado, Kibwezi, Kitale, Nairobi and Namanga regions. A total of 287 wild-caught NHPs, comprising of 115 Olive baboons (Papio anubis) and 172 Africa Green monkey-AGM (Chlorocebus aethiops) were sampled. Sera prepared from the 287 blood samples were tested for anti-dengue viral immunoglobulin gamma (IgG) antibody using enzyme linked immunosorbent assay (ELISA). Samples which were positive for anti-dengue IgG were subsequently subjected to a nested polymerase chain reaction (PCR) test with specific primers targeting the capsid and pre-membrane genes of the virus. The sero-prevalence of dengue virus exposure was 35.7% (41/115) in P anubis and 39.5% (68/172) C. aethiops. RT-PCR confirmed that 1.7% (2/115) P anubis and 2.3% (4/172) C. aethiops were positive for dengue virus. The 2 baboons and 4 AGMs were from Laikipia, Kakamega, Kibwezi and Kitale respectively. Chi-square test indicated no statistical significance in sero-prevalence (P = 0.537) results between the species. In conclusion, this study provides strong evidence of dengue virus infection among P anubis and C. aethiops in Kenya. Although this epidemiological study indicates high dengue exposure in NHPs, there was significantly low active infection as evidenced by PCR results. My conclusion is that sylvatic dengue viruses 2 circulates in non-human primates found in Kenya. I recommend that the country should strengthen its surveillance on sylvati cengue viruses which will likely become a serious public-health problem in the near future in human population that live in close proximity to animals.
Antifungal Activities of Camellia Sinensis Crude Extract on Selected Pathogenic and Mycotoxic Fungi
(2014-02-26) Sigei, Erolls Cheruiyot; Muturi, Margret; Bii, Christine
Human fungal infections pose serious medical issues. Up to now, more than a hundred thousand fungal species are considered as natural contaminants. During the last decade, the incidence of superficial and deep mycotic infections has continued to increase explosively. There is a general consensus among researchers, clinicians and pharmaceutical companies that new, potent, effective and safe antifungal drugs are needed. Majority of work has been conducted on Camellia sinensis extracts against bacterial agent’s activity but little work for antifungal activity. In this study, in vitro antifungal activities of Camellia sinensis crude extracts compared with azole group of compounds on selected pathogenic and mycotoxic fungi were determined. That was done by evaluating the difference in antifungal activities of green and black tea crude extracts having a concentration of 100mg mL-1. Quantitative bioassay was done using disc diffusion method and Minimum Inhibition Concentration was done using broth dilution methods. The fungal isolates used for bioactivity testing were yeasts; Candida famata, C. lusitaniae, C. tropicalis ATCC 750, C. parapsilosis ATCC 22019, C. glabrata ATCC 24433, C. krusei ATCC 6258 and Cryptococcus neoformans ATCC 66031; and moulds, Trichophyton mentagrophytes, Microsporum gypseum, Fusarium monilliforme, Aspergillus spp and Penicillium chrysogeneum. ATCC standard fungal strains and clinical isolates were included. Green tea crude extract showed stronger inhibitory effect against the fungal strains tested than black tea crude extract. There was a significant difference in zone of inhibitions (T=4.09, P<0.05). Zone of inhibition exhibited by green tea crude extracts (11.92±0.00mm) were higher than black tea crude extracts (8.14±0.56mm). The pattern of activity by tea crude extracts against ATCC standard fungal strains and clinical isolates strains were similar. C. famata, C. lusitaniae, C. tropicalis ATCC 750 and dermatophyte, T. mentangrophyte were inhibited by green tea crude extract (IZD≥15mm). Clinical isolates of Candida albicans (strain 4 and strain 5); Cryptococcus neoformans (strain 3, 5 and 12), showed susceptibility to Camellia sinensis green crude extracts. The MIC of Camellia sinensis crude extracts against fungal isolates tested ranged from 50 mg mL-1 to 1.6 mg mL-1. Hot green tea crude extract (mean MIC 12.25mg mL-1) had a higher MIC on clinical fungal isolates than cold green tea crude extract (Mean MIC 12.167 mg mL-1). The concentrates of aqueous Camellia sinensis crude extracts showed synergistic activity with conventional antifungal drug. However, level of synergism differed as observed in difference in inhibitory effect. The difference in inhibitory effect was significant (P<0.05). The crude tea extract restored the activity of lower concentration of antifungal, Fluconazole below MIC to susceptible breakpoints. Generally, the MFC (Minimum Fungicidal Concentration) of Mixture crude extracts were slightly higher as compared to that of green tea crude extract. These results are suggestive that addition of milk to blend the crude extracts altered the bioactive ingredients resulting to higher concentration for its MFC as compared to crude extracts alone. The studies on Camellia sinensis crude extracts (green and black) have shown remarkable antifungal activity against different strains of fungi and highlighted its significance to humans as potential health products.
The Prevalence of Human Cytomegalovirus among Pregnant Women Attending Thika Level 5 District Hospital Kiambu County, Kenya
(2014-08-19) Maingi, Mwangi Z.; Mwangi, Z. M.; Kebira, A.; Runo, Steven
Cytomegalovirus (CMV) is a member of herpesviruses. It is one of the most common causes of congenital and prenatal infections. It is a ubiquitous virus with the ability to establish latency following primary infection, and can be reactivated particularly during episodes of immunosuppression. CMV infection of pregnant women, especially in the first trimester may lead to congenital abnormalities and is often associated with serious complications, such as microcephaly, mental retardation, spastic paralysis, hepatosplenomegaly, anaemia, thrombocytopenia, deafness, and optic nerve atrophy leading to blindness in infants. The prevalence of CMV infection in developed countries is about 40% and in developing countries may be 100%.This study was done to determine the seroprevalence rate of the infection and its associated risk factors in pregnant women at Thika Level 5 District. This cross-sectional study was done in 260 pregnant women. Demographic data were collected by a questionnaire. About 3 ml of blood was taken from each patient. Aliquots of serum samples were stored at -20°C until analyzed. The presence of IgM and IgG CMV specific antibodies was assessed by enzyme linked immunosorbent assay. Data were analyzed by chi square test using EPI Info 2003 software. The prevalence rate was at 85.4% with the majority of women (77.3%) being positive for CMV-IgG in pregnancy while the rate of positive CMV-IgM infection was 8.1%. Samples that tested positive for both IgG and IgM were 20.8% and underwent avidity test to evaluate strength of the IgG antibodies. Those with avidity index >35% were 79.63% showing IgG antibody pre dominancy while those with avidity index 0.05). Adoption of CMV screening into the antenatal profile tests, health education of this virus on how it is acquired and contributing factors like personal and community hygiene were recommended as preventive approaches to congenital CMV infections.
Validation of di-electric heating as a potential food safety intervention
(2014-11) Luvanda, Maureen K.
Food and water borne diseases are leading causes of illness and death in less developed countries killing approximately 7.2 million people annually, 1.9 million of whom are children. WHO has estimated that each year 2.2 million people, including 1.9 million children, die because of diarrheal diseases. This study was intended to experimentally analyze if di-electric heating has any indirect impact on the control, management and prevention of food borne disease outbreaks in terms of reduction of bacteria infective doses and increase in antibiotic susceptibility. This included investigating the effects on bacterial colony forming unit counts before and after exposure to microwave radiation as well as investigating changes in antibiotic susceptibility patterns. Surviving colonies were also investigated for viability, growth and multiplication ability. The aim of the study was to determine if microwave radiation can be used as a food safety intervention; whereas other objectives included evaluating its effects on growth and multiplication of bacteria. reduction of bacterial infective doses, measuring appropriate time (in minutes) required for radiation to achieve the desired results, recording the most effective power at which radiation was most likely to achieve the desired results and lastly observing if there are any significant changes as a result of exposure to microwave radiation when it comes to antibiotic susceptibility patterns. Various food samples (n=73) were artificially contaminated by food borne disease pathogens (Escherichia coli, Staphylococcus aureus, Shigella flexneri and Salmonella paratyphi) and were exposed to microwave radiation at different powers (P-OO, P-20. P-40, P-60, P-80) and time periods (2 rnin, 4 min, 6 min, 8 min).The degree of inactivation was estimated by measuring the colony forming units formed in culture before and after exposure of samples to radiation (Pre-test Post-test experimental design). The laboratory methods used were mainly culture techniques and biochemical tests. Data entry was done in Microsoft Excel after which data analysis was carried out using Statistical Package for Social Science (SPSS V 13) software. The results showed that generally microwave radiation produced a 1-2 log reduction when artificially contaminated food samples were processed. Initial seeded bacterial numbers (>3.0 x 104 ) were reduced significantly I' (P<0.05) however, this was not the same case when it came to antibiotic susceptibility patterns. The most effective time-power combination for bacteria inactivation was 8 minutes at P-80 where bacteria numbers were very low «1.0 x 10\ whereas the least effective time-power combination for bacteria inactivation was 2 minutes at P-20 where bacteria numbers remained at (>3.0 x 104).The study shows that microwave radiation reduces infective doses of bacteria by 96.67% but does not affect their antibiotic susceptibility patterns. Indications of any other effects of microwave radiation on the microbial agents were als noted. This study will serve as a guide to policies implemented on management and control of food and water borne infections. Traditional methods for improvements in food safety are still being utilized however the number of food and water borne disease incidences and antibiotic-resistance cases continue to rise. This therefore means that prevention rather than cure is important now more than ever and so modern methods or practices such as the microwave radiation that promote food hygiene should be explored and encouraged.
Evaluation of Serum Agglutination and Rose Bengal Tests against Immunofluorescence Antibody Assay in Diagnosis of Brucellosis in Narok and Kiambu County, Kenya
(Kenyatta University, 2015) Nyagah, Susan, Nyawira
Brucellosis in human is characterized by non specific signs and often misdiagnosed as malaria, typhoid, rheumatic fever and pyrexia of unknown origin. Definitive diagnosis of brucellosis requires isolation of Brucella '\PP.but chances of successful isolation decrease as the disease progresses. In Kenya, the current routine diagnostic test, Serum Agglutination Test (SAT) has a major drawback in that its not suitable for patient follow-up, since titers can remain high for a prolonged period. The alternative test, Rose Bengal Test (RBT) is not reproducible as it is very sensitive to vaccinal antibody limiting its use in vaccinated animals. Immunotlourescence antibody assay (IF A) has been described as the gold standard for diagnosis ofbrucellosis, To date there is no study in Kenya that has examined performance of the two ronrme tests STand RBT in the face of IFA The objective of this study was to evaluate SAT and RBT against IFA and to estimate the prevalence of brucellosis in Kiambu and Narok counties by the three diagnostic methods. Human blood and unpasteurised milk samples were collected between December 2009 and August2010 in Kiambu and Narok counties. A total of 250 human serum and 250 milk samples were collected from arok and Kiambu counties. Human serum sample was obtained from Kijabe referral hospital and other neighbouring health facilities. Antibodies to Brucella spp. were screened using SAT and RBTin the hospital and later transported to Kabete Veterinary Laboratory for IFA Milk samples were collected from milk vendors packed in cool boxes and transported to Kabete Veterinary Laboratory where screening was done using one step bovine Brucella antibody rapid test. All samples were subjected to IFA to determine the prevalence of brucellosis. The overall prevalence ofbnlcella antibodies by IFAas a gold standard was 3.2% and 70.4% and 14.4% and 54.4% in human samples and milk samples in Kiambu and Narok respectively. Prevalence by SAT was 10.4% and 77.6% and 8.3% and 80% by RBT in Kiambu and Narok respectively. Comparison of SAT and IFA gave PPV (63%), NPV (16.4%), sensitivity (75%) and specificity (74%). Comparison of RBT and IFA gave sensitivity (69.56%), specificity (72.78%), PPV(59.81 %) and NPV(19.58%). Comparison of One step bovine brucella rapid agglutination test and IFA gave sensitivity (16.5%), specificity (53.0%), PPV (19.76%) and NPV (52.43%). Prevalence was higher in females (68%) than males (32%) in Narok particulary 21-30 years. Both SAT and RBT should be scrapped as they lack sensitivity and specificity. IFA which is accurate, sensitive, specific, rapid should be recommended as the diagnostic test of choice. People handling animals and their products are at higher risk of infection.
Isolation and characterization of antibiotic producing thermophilic bacillus in selected hot-springs along lake Bogoria, Kenya
(2015) Torome, Tom Kintet
Extreme habitats are increasingly being recognized as sources of secondary metabolites which provide an encouraging source for development of novel natural pharmaceuticals. Saline and hot water lakes are now becoming appreciated as rich and untapped reservoirs of extremophilic microorganisms with previously uncharacterized functions. Bacilli, a large homogeneous group of bacteria that survive in a wide range of environmental conditions, is one such microorganism. Formation of resistant spores allows it to survive in high temperatures zones where other organisms cannot. The objective of this study was to isolate, identify, and characterize Bacillus organisms with the potential to produce secondary metabolites with antibacterial properties from the hot springs of Lake Bogoria. Eighty samples were collected from surface and sediment waters of selected hot springs and inoculated directly into nutrient broth in universal bottles. Samples were initially incubated at 45oC for 48 hours and growth was examined by checking for turbidity. Out of the 80 bottles inoculated with sample, thirty three exhibited growth. Subsequent streaking on nutrient agar and Gram staining showed that nine of the isolated organisms were Gram-positive rods, twenty were Gram-negative rods, and four were Gram-positive cocci. Only Gram-positive rods were processed further. Biochemical characterization revealed that all nine processed isolates were catalase-positive and non produced gas upon sugar fermentation. They however had varying results for indole, motility, lactose fermentation, glucose fermentation and H2S production. Antibacterial profiling was performed using ATCC organisms: Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Escherichia coli 35218, and Pseudomonas aeruginosa ATCC 27853. Isolates D1, D5, D8, D22 and S15 inhibited growth of Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 which was shown by clear zones of inhibition around the discs. No inhibition was noted against S. aureus ATCC 29213 and E. coli 35218. Phylogenetic analysis of amplified 16S rDNA gene showed that D1, D5, D8, D22 and S15 formed close phylogenetic clusters with known members of Bacillus organisms with a (88-99%) sequence identity. The current study shows the presence of thermophilic Bacillus species, which are producers of biomolecules with antibacterial properties within the hot springs of Lake Bogoria. Characterization of the antibacterial compounds produced will be useful for the discovery of novel antimicrobial substances that are effective against wide range of pathogens.
Morbidity, Risk Factors, and Flea Species Responsible For Tungiasis in Selected Villages in Kisumu County, Kenya
(Kenyatta University, 2015) Okoth, A. A.
Tungiasis is a neglected disease that afflicts four million Kenyans. To control its spread, there is a high demand for sustainable disease control strategies in rural endemic areas. The objective of this study was to determine the prevalence of tungiasis, associated risk factors, and the species of fleas causing it in rural western Kenya. A crossectional study was done in Otwenya Location, Maseno Division between August and September 2013. Seventy eight (78) randomly selected households were visited all household members (415) checked for tungiasis. Structured questionnaires were used to collect demographic and environmental data from household heads, neosomic and free-living fleas collected in houses and characterised using taxonomy and Polymerase Chain Reaction (PCR). Data analysis was done using the Statistical Package for Social Scientists (SPSS) version 17. Means and frequencies were calculated and the Fishers exact test and logistic regression with backward elimination used to identify risk factors for tungiasis. Tests for statistical significance were done at 95% confidence interval (CI). Participants were between 1 and 83 years old (females (52.8%) and males (47.2%)). In 39.7% of households, at least one case of tungiasis was found. The overall prevalence was 19.5%, peaking among 5-9 years olds (37.7%) and the elderly (60+ years) (43.8%). Tungiasis cases had between one and 53 lesions. The mean parasite load was 8.3 (males (9.6±1.6) and females (6.5±0.98), with pathological manifestations for tungiasis such as deformed toenails identified on 71.6% of cases. Approximately 19.8% had signs of bacterial infections. Living in houses with sandy flooring was a major risk factor for tungiasis (OR=11.1, p=0.01). Living in houses with animals such as dogs and cattle (OR=22.8, p=0.01) and Overcrowding in houses (OR=3.5, p=0.04) were other risk factors for the disease. Stigma and negligence played a major role in the establishment and proliferation of tungiasis in the study area. Diverse species of fleas were trapped in study houses. However, Tunga penetrans was the only species causing tungiasis. Otwenya Location is a tungiasis endemic area. Sensitisation of residents on its control, developing outreach programs that dispel negligence and stigma, and eliminating modifiable risk factors in homes can help to lower its burden in the area
Bacterial Flora as Determinants of Ventilator Associated Pneumonia in Intubated Patients in an Intensive Care Unit
(Kenyatta University, 2015-01-14) Nkoroi, Beatrice Nkirote
Infection is a major cause of morbidity and mortality in intensive care units (ICUs) worldwide. Of these, ventilator associated pneumonia (VAP), a common complication of endotracheal intubation and subsequent mechanical ventilatory support in critically ill ICU patients is a major concern for intensivists and other health care players. The issue of concern is whether the microbial agent present in the respiratory tract of patients at the time of intubation is likely to cause VAP in the intubated and mechanically ventilated patient. Studies done internationally have established a profile of some of the common bacterial agents responsible for VAP, however no studies have been done in Kenya to profile the common pathogens responsible for VAP and therefore, relatively little information is available about the Kenyan epidemiology of such infections. The purpose of this study was to provide an up-to-date picture of the demographic characteristics of patients who develop VAP, the prevalence of VAP and the common bacterial causative agents and the antimicrobial sensitivity patterns in a mixed medical – surgical ICU of the Aga Khan University Hospital, Nairobi Kenya. One hundred patients admitted into the ICU who were intubated during the study period were enrolled. Endotracheal aspirates were taken at the time of intubation and repeated when a clinical suspicion of VAP was made. Data was collected using preprinted case report forms and analyzed using chi-square and Pearson’s correlation as appropriate using the statistical software for social sciences (SPSS) version 17. Data was presented in tables, histograms, and other pictorial presentations as appropriate. The mean age of the patients was 55.86 years (95 % CI: 52.15, 59.67). Males were 57(57 %) and females were 43 (43 %).Most of the patients were admitted to ICU via A and E (35 %). Patients with medical conditions were (57) 57 %, those with surgical conditions were 25 (25 %) while 18 (18 %) had both a medical and surgical condition. A total of 78 (78 %) patients had chronic comorbidities while twenty two (22) patients did not have any comorbidity. Among the 78 who had comorbidities only 9 (12 %) had respiratory related comorbidities. A total of 21 (21 %) patients had a history of respiratory infection at the time of admission into ICU. Sixteen patients developed VAP, giving the prevalence of VAP at the AKUH, N ICU of 16 % (95 % CI: 14.0-23.6 %).The most common bacterial pathogens isolated from the patients with VAP were Acinetobacter baumanii (37.5 %), Pseudomonas aeruginosa (12.5 %), Klebsiella pneumoniae (6.3 %). Candida albicans was isolated in 12.5 % of patients with VAP. Thirty two percent (32 %) of the pathogens isolated demonstrated high resistance and MDR patterns to the antibiotics commonly prescribed in this ICU. Meronem (26 %), Tazopipril and Rocephine each at 12 % were the most commonly prescribed antimicrobial agents. Further studies are recommended to establish the susceptibility patterns of the individual antimicrobial agents and evaluate the clinical outcomes of patients with VAP
The Cytokine Profile and Prostate Specific Antigen Levels in Prostate Cancer Patients at Kenyatta National Hospital
(Kenyatta University, 2015-08) Mwirigi, Liza Kiende
Prostate cancer (PCa) is Kenya‘s most frequently diagnosed cancer of men with an incidence rate of 32 per 100,000 and mortality rate of 12 per 100,000. It is also the leading cancer in terms of incidence and mortality in men from Africa and the Caribbean and the numbers may double by 2030. Prostate Specific Antigen (PSA) is the main diagnostic biomarker used for screening patients currently. Biopsy histology is the only confirmatory test, but it is invasive and tedious to perform. There is an urgent need to develop other accurate biomarkers that can be used for screening and diagnosis. Cytokines have potential but have not been extensively studied as biomarkers. The objectives of this study were to evaluate the cytokine profile, determine the PSA levels in PCa patients and the risk factors that lead to the development of PCa. The study profiled six cytokines (interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon gamma (IFN-Gamma) and tumor necrosis factor alpha (TNF-Alpha) and the total PSA levels in the serum of 45 PCa patients, 7 benign prostate enlargement (BPE) patients and 7 healthy males as controls. Cytokines were measured using a highly sensitive cytometric bead array technique. PSA levels were determined using enzyme linked immunosorbent assay (ELISA) technique. A questionnaire was used to determine the demographic and risk factors that lead to the development of PCa. Statistical analysis was performed using statistical package for social sciences (SPSS) version 21. Analysis of variance (ANOVA) was used to compare the means of the cytokines and PSA levels in the different Gleason scores of the PCa patients. Pearson correlation was used to correlate the six individual cytokines and PSA levels. Statistical significance was set at the level of p<0.05 for both the ANOVA and the Pearson correlation test. The results showed that serum levels of all cytokines and PSA were significantly (p<0.05) higher in the PCa and the BPE patients. The PCa patients with a Gleason score of 8-10 had the highest levels of all the cytokines followed by those with a Gleason score of 5-7, then those with a Gleason score of 2-4. TNF-Alpha and IL-6 were the cytokines with the highest levels while IL-4 and IL-10 had the lowest levels. The cytokine levels among the BPE patients were less than those of the PCa patients. The PSA levels were also significantly higher (p=0.002) in the BPE and PCa patients and they were significantly correlated (p=0.01) with the six cytokines in the two study groups. Kenyan men who are above 50 years are at risk of suffering from PCa. These results indicated that cytokine profiles in PCa patients are distinct by the stage of the disease. TNF-Alpha, IL-6, IFN-Gamma, IL-2 and IL-4 may be potential early diagnostic biomarkers for prostate cancer. The current data forms a basis for further investigations.
Immunogenicity of 10-Valent Pneumococcal Conjugate Vaccine among Infants at Mbagathi District Hospital, Kenya
(Kenyatta University, 2015-08) Walekhwa, Michael Nyongesa
Pneumococcal diseases are responsible for killing at least one million children under the age of five every year: over 70% of these deaths are in developing countries. This total is greater than that due to malaria, AIDS and measles combined. Streptococcus pneumoniae has over 90 serotypes, out of which eight are included in the 10-valent Pneumococcal conjugate vaccine (Synflorix) currently in use in Kenya. Pneumococci are highly diverse and serotype prevalence is dynamic based on age, time period and geographical area. There is no published data on serotypes found in Nairobi and other areas in Kenya except the coastal region. It is therefore possible that these serotypes are different from those included in the vaccine, subjecting the immunogenicity of the vaccine among Kenyan infants to doubt. Since its launch for free public use in February, 2011increasing number of children has been immunized, yet, it was not clear to what extent the vaccine is protective. This study therefore evaluated the immunogenicity of the vaccine by measuring serum concentration of IgG antibodies among infants immunized with PCV-10 at the Mbagathi District Hospital (MDH). It also investigated; the vaccine herd effect on study infants biological mothers and factors that affect the vaccine immunogenicity. It was a cross-sectional study where infants that completed 3-doses of PCV-10 had IgG antibodies to serotype-specific capsular polysaccharide measured by enzyme-linked immunosorbent assay. The majority (76.1%) of the infants who had completed the required dose of the pneumococcal conjugate vaccine had IgG serum titres of between 0.34mg/dl to 0.36mg/dl. Thirteen percent of infants had 0.30mg/dl to 0.33mg/dl serum concentration of PD specific IgG antibodies. The remainder of the studied infants had IgG antibody titres ranging between 0.25mg/dl to 0.29mg/dl and ≤0.25mg/dl respectively. Also, the total score for serum concentration of PD specific IgG among study infants biological mothers was between 0.34 – 0.36 (61.6%), which meets the threshold set by WHO. This study also found out that there was multi-collinearity between IgG antibody levels (mg/dl) for infants and vaccinated infant’s biological mothers and several variables. These variables included: Use of alcoholic drinks by infants’ biological mother during pregnancy (r =.595,p ≤ 0.05); Maternal diet during pregnancy (r =.137,p ≤ 0.05); Breast feeding frequency (r =.220, p ≤ 0.05); Gap with other children (r =.133, p ≤ 0.05); Child hospitalization (r =.131, p ≤ 0.05); Chronic illness (r =.154, p ≤0.01); and IgG antibody levels (mg/dl)for infants (r =.675,p > 0.05). The 10-valent pneumococcal conjugate vaccine is immunogenic against Pneumococcal Disease four weeks after completion of three doses among infants attending child welfare clinic at Mbagathi District Hospital, Ken
Prevalence of HIV/HCV co-infection amongst HIV serodiscordant couples in Thika, Kenya
(Kenyatta University, 2016-04) Waweru, Susan Wairimu
Immune defects caused by Human immunodeficiency virus (HIV) and Hepatitis C virus (HCV) co-infection could alter the course of secondary infection and dysregulated innate immune responses could contribute to a more rapid disease progression. The prevalence of HCV and HIV co-infection in Kenya varies with different risk groups. Currently, there is parity of data on the prevalence of HIV/HCV co-infection in Kenya. It is also not known how the CD4+ and CD8+ counts correlate with HIV viral load in HIV/HCV co- infected individuals. This study focused on determining the prevalence of HCV and how the CD4+ and CD8+ counts correlate with HIV viral load in HIV/HCV co-infected individuals amongst HIV serodiscordant couples in Thika. Plasma samples from HIV serodiscordant couples that had been collected over a period of 2 years (2006-2008) and stored at -20° C at the Clinical Trials Research Laboratory (CTRL) were used. All the samples from both HIV negative and positive participants were tested for HCV antibody using Murex anti-HCV (version 4.0) microelisa kit. HIV RNA viral load and CD4+/CD8+ counts were obtained from already archived data for the participants that were co-infected with HIV and HCV. A total of 385 samples from HIV serodiscordant couples were used in this study; 196 HIV positive samples and 189 HIV negative samples. Data was analyzed using mean (standard deviations), medians (inter-quartile range), Pearson correlation, Pearson chi square test, McNemar chi square test, ranksum test and t-test as appropriate using Stata version 12.1. Results obtained from this study showed that the mean age of the participants was 34.2 years. 192 of the participants were male (49.9%) and 193 (50.1%) were female. Overall, 13 (3.4%) of the participants were positive for HCV, most of whom 11(5.6%) were HIV positive, p= 0.013. There was no statistically significant difference between HCV infection on the basis of gender, 7 (3.6%) vs. 6 (3.1%), p= 0.785. A negative correlation was observed between HIV Viral load and CD4+ T cells amongst HIV/HCV co-infected participants without statistical significance; r= 0.600, p= 0.070. A positive correlation was observed between HIV Viral load and CD8+ T cells amongst HIV/HCV co-infected participants without statistical significance r= 0.4525, p= 0.162. There was a statistically significant higher median (IQR) HIV Viral load (copies/mL) at baseline among those positive for HIV/HCV co-infection than those who were HIV mono-infected, 89775 vs. 10695, P= 0.0436. Those positive for HIV/HCV co-infection had a lower median CD4+ (IQR) than those who were HIV mono-infected, however without statistical significance, 383 vs. 499, p=0.0772. Those positive for HIV/HCV co-infection had a higher median CD8+ (IQR) than those who are negative for HIV/HCV co-infection, however without statistical significance, p=0.2250. None of the serodiscordant couples were both found to be positive for HCV indicating that HCV may not have been transmitted sexually. A longitudinal study is recommended using a higher number of participants and also inclusion of more laboratory tests and parameters to give a better understanding of the effect of HIV/HCV co- infection on disease progression amongst participants.
Evaluation of safety and therapeutic potential of environmental waste water lytic bacteriophage against multidrug resistant staphylococcus aureus (MDRSA) in BALB/c mice
(Kenyatta University, 2016-04) Oduor, Joseph Michael O.
Methicillin resistant Staphylococcus aureus (MRSA) pose a great threat to the global public health. Control of these bacteria has become difficult due to acquisition of resistance against even some of the best antibiotics. Thus, phage therapy could be the better alternative as they are easy to isolate and produce in mass within a short time. However, phage therapy has been a subject of debate over the years but recently there has been a renewed interest due to their proved therapeutic potential and have, therefore, found commercial application in some countries. Thus, this study was specifically designed to evaluate the efficacy of phages against MRSA both in vitro and in vivo. A litre of environmental waste water and sewage samples were collected around the county of Nairobi. The MRSA isolates were obtained from environmental waste water and sewage samples from Nairobi and its environs and evaluated for drug resistance using antibiogram test. In addition, lytic phages were isolated from these samples too. Thereafter, the in vitro efficacy of the phages against MRSA was done by spot assay and tube culture tests. Only the most virulent phage isolate was used for in vivo efficacy study which involved six groups of mice of n=5 per group (BALB/c mice; both sex). The first three groups acted as controls (group 1=only physiological saline, group 2=MRSA bacteria only and group 3=phage only) while the remaining groups were used for efficacy studies using a dosage of 108 CFU/ml for MRSA bacteria, 108 PFU/ml for phage and clindamycin at 8mg/kg. The efficacy study groups (groups 4-6) were first infected by MRSA and observed for 3 days before treatment with either antibiotic (group 4), or phage (group 5) or phage + antibiotic (cocktail therapy group 6). The mice were then observed for an additional 7 days. During the entire 10 days of observation blood samples were collected daily for bacteremia level determination before being euthanized. Different organs including the liver, brain, kidney and lungs were harvested for histopathological studies. All studies were done in accordance with the IPR Institutional Ethical Committee approved protocols. Antibiogram test indicated that MRSA isolated was a multidrug resistant strain (Figure 1). While the in vitro test showed the virulence of the phage isolates after 24 hrs culture at 370C. Toxicity test showed that phages were safe. There was no significant difference in survival rates between phage infected group and non-infected control group (p>0.05). Bacteremia was significantly lower in phage treated group as compared to other treatment groups and bacteria non-treated group (p<0.001) (Figure 2). In addition, pathological results show that phage prevented organ damage by the bacteria (Figure 3). Thus, a single dose of phage was more effective than other therapeutic agents used in the study. Results of this study show that phage therapy is safe and its application should be considered for the treatment of multidrug resistant bacterial infections.
Plasma interferon-gamma, interleukin-10 and adiponectin levels in HIV-1 and tuberculosis coinfected injection drug users at Bomu Hospital, Mombasa, Kenya
(Kenyatta University, 2016-06) Kiboi, Nathan; Were, Tom; Juma, Gerald
Sub-Saharan Africa accounts for high tuberculosis cases that result from widespread HIV infections, which is exacerbated by injection substance use. Immunologically, HIV critically impairs cell-mediated host responses to Mycobacterium tuberculosis. IFN-γ, IL-10 and Acrp30 are key mediators of systemic inflammation. Although circulating IFN- and IL-10 levels are increased, Acrp30 levels are lowered and associated with disease severity among HIV and TB co-infected non-susbstance users. In contrast, circulating IFN- and Acrp30 levels are decreased while IL-10 levels are upregulated among injecting heroin addicts. However, no studies to date have reported on these cytokine profiles among Kenyan HIV-1 and TB co-infected injection drug users. This study, therefore, investigated plasma IFN-γ, IL-10 and Acrp30 levels among IDUs, and their association with CD4+ T cell counts, HIV-1 viral load and BMI. A cross-sectional study was conducted from August, 2012-November, 2013 using 138 participants recruited at Bomu hospital; a major centre for rehabilitation of drug and substance users in Mombasa County. Following informed consent, IDUs were enrolled through respondent driven sampling, snowball and makeshift methods while convenience and purposive sampling were used for recruiting the control group. IDUs and controls were screened for HIV and TB respectively through Determine™ and Bioline™ rapid tests, and Ziehl Neelsen stained sputum smears. Subsequently, the study participants were categorised into: HIV-1/TB coinfected ART-naive (n=9) and -experienced (n=27); HIV-1 mono-infected ARTnaive (n=26) and -experienced (n=13); TB mono-infected (n=21), HIV-1 negative and TB uninfected (n=25) IDUs and controls (n=17). Demographic, drug use information and physical measurements were recorded using assisted interviews. EDTA venous blood samples were collected and used for preparing plasma and enumerating CD4+ T cell counts. Frozen plasma samples were used for determining cytokine concentrations, and HIV-1 viral load. CD4+ T cell counts were enumerated using flow cytometry; cytokine levels were measured using a sandwich ELISA technique, while HIV-1 viral load was determined by RT-PCR, respectively. Across-group comparisons in continuous data were performed using Kruskal Wallis followed by post-hoc Dunn’s tests. Plasma IFN-γ (P<0.0001), IL- 10 (P<0.0001) and Acrp30 (P=0.006) levels differed significantly across groups. IFN-γ levels were high in co-infected ART-naive (P<0.001) and -experienced (P<0.001), and HIV-1 mono-infected ART-experienced (P<0.001) IDUs relative to healthy controls. IL-10 levels were elevated in uninfected IDUs (P<0.001) compared to healthy controls. Acrp30 levels were lower in TB mono-infected (P<0.01) relative to controls. IFN-γ/IL-10 ratio varied across-groups (P<0.0001) and higher in co-infected ART-naive (P<0.001) and -experienced (P<0.001), and HIV-1 mono-infected ART-experienced (P<0.001) compared to uninfected IDUs. The IFN-γ/Acrp30 ratio also differed across groups (P<0.0001) with HIV-1 mono-infected ART-experienced (P<0.001), and co-infected ART-naive xii (P<0.001) and -experienced (P<0.001) IDUs exhibiting higher ratio relative to uninfected IDUs. CD4+ T cells correlated inversely with Acrp30 (=-0.717, P=0.030) levels in TB mono-infected IDUs whereas BMI correlated positively with Acrp30 (=0.523, P=0.022) among co-infected ART-naive IDUs, respectively. Altogether, circulating IFN-, IL-10 and Acrp30 production is altered in ART-naive and -experienced HIV-1 and TB co-infected IDUs, suggesting a role as disease markers in HIV and TB co-infection among IDUs.
Correlation between an early 6 and early 7 oncogenes detection method and conventional cytology for cervical cancer screening at Kenyatta National Hospital, Kenya
(Kenyatta University, 2016-10) Muyabwa, Cosmas
The oncogenic potential of human papilloma virus (HPV) early genes E6 and E7 is of interest in HPV testing for cervical carcinoma. The current study included women less than 40 years attending the Kenyatta National Hospital family planning clinic, with an objective of comparing the performance of cytological tests done in Kenya with the Pretect SEE assay developed by Norchip AS (Norway). Two hundred and four (204) samples were obtained in which the HPV E6 and E7 HPV mRNA was evaluated using the Pretect SEE assay for detection of high risk HPV types 16, 18 and 45. The HPV 35 and 52 may not be involved as the cause of invasive cervical cancer in Kenya. The real-time nucleic acid sequence-based on amplification was also included. The Pap test and cytological analysis were done locally at the Kenyatta National Hospital and the results were recorded and compared to the Pretect SEE results. Three (3) out of two hundred and four (203) women were positive with Pretect SEE (samples that expressed E6 and E7 HPV oncogenes), whereas only one sample(1) out of two hundred and three 203 samples was positive with Pap test. One sample (1) was rejected due to lack of cells. The overall diagnostic prevalence of HPV was 1.47% (3/203) after testing with pretect SEE and 0.5% (11203) after testing the pap smears. The Pretect SEE showed a specificity of 100% and a sensitivity of 100% using Pap smear as Gold standard. The PreTect SEE showed a positive predictive value (PPV) of 33 % and a 99% negative predictive value using Pap smear as the gold standard. The Pap smear showed a specificity of 100% and a sensitivity of 33% when using PreTect SEE as the gold standard. If all the PreTect SEE contains CIN2+ cases, the sensitivity, specificity, PPV and NPV would be 100%. In conclusion, the Pretect SEE™ assay was found to be more sensitive than the Pap smear. Therefore the use of PreTect SEE as a primary screening method with a high coverage rate may reduce the incidence of cervical pre-cancer to a minimum in Kenya. Using a very sensitive and specific method in a very representative female population in Kenya has strongly indicated that the prevalence of cervical cancer in Kenya may be lower due to false positive and lack of differentiation between transient and transforming infections. The estimated number of annual cervical cancer cases may be around 1000 and not 4800 as indicated by the IeO HPV Information (2012). The estimated prevalence of HPV 16 and/or HPV 18 DNA (2012) reported by ICO HPV Information Centre among cytological women in Kenya, may be less than 2,5%and not 9.1%. Around 40% of the HPV 16 and 18 HPV DNA in cytological normal cases may not express E6 and E7 mRNA. This also show that the prevalence of HIV in the whole Kenya population may not be 6.2% but rather less than 2%. The study showed that it is important to perform national screening in more than 3.2% of typical national population's in Africa.
Comparative Study of Hepatitis B and Hepatitis C Viruses among HIV-1 Infected Intravenous Drug Users and Non-Users In Mombasa County
(Kenyatta University, 2017) Kinya, Caroline Mercy
HIV/AIDS is a debilitating disease associated with high mortality and morbidity globally. In Kenya, it is the major cause of mortality across all gender and age groups, in effect, putting a huge demand on the healthcare system and the economy. The HIV 1 positive population faces major challenges such as the drug resistance, severe hepatic coupled with immunological deficiencies and toxicity. The problems are aggravated by co-infection with blood borne diseases, varied responses to the infection and therapy among vulnerable groups. This study purposed to determine and compare; the CD4 counts, HIV viral loads, liver enzyme markers and the prevalence of viral hepatitis among the HIV- 1 positive IDUs and NDUs. A cross-sectional study was carried out in Mombasa County. A structured questionnaire was used to collect information on HIV AIDS, Hepatitis and social demographics. Blood samples were collected, screened and analyzed for HIV, HBV and HCV, CD4+ cell, HIV viral load and liver enzyme markers. Pearson’s Chi square, Student T test and one way Anova were used to analyze data. A P value ≤ 0.05 was considered statistically significant. A total of 200 participants consisting of 78 males and 122 females were recruited. Sero-prevalence of Hepatitis B and Hepatitis C was at 16% and 20% respectively among the IDUs and 11% and 8% respectively among the NDUs. The sero-prevalence of HIV+HBV+/HCV infection among IDUs and NDUs was 6% and 4% respectively. The liver enzyme markers (Alkaline phosphate, Aspartate aminotransferase and Alanine transferase) were markedly elevated among the co-infected participants than the mono-infected in both the NDU and IDU groups. IDUs showed elevated mean liver enzymes than the NDUs. The IDUs had a lower mean CD4+ cell count of 350.2 (±225.27) cells/μl than the NDUs 485.9 (±243.38) cells/μl (P˂0.0001). Participants who were co-infected showed remarkably low mean CD4+ cell counts of 192. 91(±84.08) cells/μl than the mono-infected with mean count of 536.79 (±218.76) cells/μl (P=˂0.0004). A statistically significant high mean viral load of 153392.97(±395699.65) copies/ml was reported among co-infected participants than the mono-infected at 2435.05(±5596.88) copies/ml (P˂0.0001) across the study population. The study established that the co-infection rates with HBVand HCV was higher among the IDUs than the NDUs. The liver profile indices indicated elevated liver enzymes among IDUs than NDUs. Co-infected participants had statistically significant higher liver enzyme markers than the mono-infected. Immunologically, CD4+ cell counts were lower among IDUs than the NDUs. Co-infected individuals had a lower mean CD4+ cell count than the mono-infected. IDUs had a statistically significant higher HIV viral load than the NDUs. The co-infected also indicated a statistically significant higher mean viral load than the mono-infected. This study proposes routine baseline screening of HBV and HCV for IDUs and NDUs owing to the high frequency of co-infections. A people driven campaign is also necessary to create awareness on the effects of the use of substances of abuse in relation to viral infections and treatment. Also the campaign is necessary to create awareness on the HBV vaccination.
Expression of integrin markers on anti- malaria effector cells in the placenta of post partum women in Western Kenya
(2017-06) Mwalimu, Stephen M.
Malaria in pregnancy has been linked to utero-placental vascular insufficiency and fetal fatalities. The immunity to placental malaria may be mediated in part by monocytes and T cells that accumulate in the placenta intervillous space. The immunological basis for retention of antimalarial effector cells in the intervillous space and the role of leucocyte integrins in the intervillous space is not well understood. Leucocytes use surface receptors to mediate binding on endothelial villous walls. Among these receptors, integrins play a crucial role in leucocytes tethering and extravasation. The surface expression of integrins and intracellular adhesion molecule-1(ICAM- 1) on T cells and monocytes in reduced incidence of placental malaria in multigravidae has not been studied. The objective of this study was to analyse the integrins and compare their expression patterns on peripheral and placental T cells and monocytes. Placental and peripheral leucocytes were processed from whole blood peripheral and intervillous blood. The washed leucocytes were stained with specific monoclonal antibodies and analysed by flow cytometry for ICAM-1, very late leucocyte antigen-4 (VLA), leucocyte function associated antigen-1(LFA-1) and macrophage antigen-1 (MaC-1). Microscopy was used to determine peripheral and placental malaria infections, while parasites density was calculated from the number of malaria parasites per 300 leucocytes and converted into parasites per microliter of blood. Statistical analysis was done using Graph Pad Prism version 5.0. Differences in the expression of integrins between two groups and within cell population were performed by students paired t-test. Statistical significance was considered for probability values less than 0 .05 (˂ 0.05).The results indicated the expression pattern of most integrins studied was altered in placental blood in presence of malaria parasites. In placental malaria ICAM-1 and VLA-4, MaC-1 and LFA-1 expression in placental monocytes was significantly up regulated (p < 0.0001). Surface expression of ICAM-1, VLA-4 and LFA-1 was significant high on peripheral T cells (P < 0.0001) in presence malaria parasites except for MaC- 1. Gravidity and parasite density in the intervillous and peripheral blood did not seem to affect the expression pattern of these integrins and ICAM-1. The results from this study suggest that leucocyte integrins activation in the placental blood is inducible by accumulation of infected red blood cells in the intervillous blood. However, the expression pattern is different on peripheral and intervillous T cells and monocytes. The study also demonstrated parasite density and gravidity does not affect the expression patterns of leucocyte integrins and ICAM -1 on peripheral and placental T cells and monocytes. Therefore to develop anti-adhesion vaccines and therapeutics to reduce malaria in pregnancy there is need to understand the role of leucocyte integrins in the pathogenesis placental malaria.
Comparative Study of Hepatitis B and Hepatitis C Viruses Among Hiv-1 Infected Intravenous Drug Users and Non-Users in Mombasa County
(Kenyatta University, 2017-09) Kinya, Caroline Mercy
HIV/AIDS is a debilitating disease associated with high mortality and morbidity globally. In Kenya, it is the major cause of mortality across all gender and age groups, in effect, putting a huge demand on the healthcare system and the economy. The HIV 1 positive population faces major challenges such as the drug resistance, severe hepatic coupled with immunological deficiencies and toxicity. The problems are aggravated by co-infection with blood borne diseases, varied responses to the infection and therapy among vulnerable groups. This study purposed to determine and compare; the CD4 counts, HIV viral loads, liver enzyme markers and the prevalence of viral hepatitis among the HIV- 1 positive IDUs and NDUs. A cross-sectional study was carried out in Mombasa County. A structured questionnaire was used to collect information on HIV AIDS, Hepatitis and social demographics. Blood samples were collected, screened and analyzed for HIV, HBV and HCV, CD4+ cell, HIV viral load and liver enzyme markers. Pearson’s Chi square, Student T test and one way Anova were used to analyze data. A P value ≤ 0.05 was considered statistically significant. A total of 200 participants consisting of 78 males and 122 females were recruited. Sero-prevalence of Hepatitis B and Hepatitis C was at 16% and 20% respectively among the IDUs and 11% and 8% respectively among the NDUs. The sero-prevalence of HIV+HBV+/HCV infection among IDUs and NDUs was 6% and 4% respectively. The liver enzyme markers (Alkaline phosphate, Aspartate aminotransferase and Alanine transferase) were markedly elevated among the co-infected participants than the mono-infected in both the NDU and IDU groups. IDUs showed elevated mean liver enzymes than the NDUs. The IDUs had a lower mean CD4+ cell count of 350.2 (±225.27) cells/μl than the NDUs 485.9 (±243.38) cells/μl (P˂0.0001). Participants who were co-infected showed remarkably low mean CD4+ cell counts of 192. 91(±84.08) cells/μl than the mono-infected with mean count of 536.79 (±218.76) cells/μl (P=˂0.0004). A statistically significant high mean viral load of 153392.97(±395699.65) copies/ml was reported among co-infected participants than the mono-infected at 2435.05(±5596.88) copies/ml (P˂0.0001) across the study population. The study established that the co-infection rates with HBVand HCV was higher among the IDUs than the NDUs. The liver profile indices indicated elevated liver enzymes among IDUs than NDUs. Co-infected participants had statistically significant higher liver enzyme markers than the mono-infected. Immunologically, CD4+ cell counts were lower among IDUs than the NDUs. Co-infected individuals had a lower mean CD4+ cell count than the mono-infected. IDUs had a statistically significant higher HIV viral load than the NDUs. The co-infected also indicated a statistically significant higher mean viral load than the mono-infected. This study proposes routine baseline screening of HBV and HCV for IDUs and NDUs owing to the high frequency of co-infections. A people driven campaign is also necessary to create awareness on the effects of the use of substances of abuse in relation to viral infections and treatment. Also the campaign is necessary to create awareness on the HBV vaccination.
Determination of Urinary Tract Infections aand Antibiotic Sensitivity among Non-Insulin Dependent Diabetes Mellitus Patients Visiting Kisii Teaching And Referral Hospital, Kenya
(Kenyatta University, 2018) Mogaka, Mageto Vincent
People with non-insulin dependent diabetes mellitus have been found to be prone to urinary tract infections. There is a wide gap of information in developing countries regarding the prevalence and antibiotic sensitivity of the pathogens causing urinary tract infections in diabetic patients. Developed nations also face a relatively huge burden of managing urinary tract infections among non-insulin dependent diabetics. The objective of this study was to determine the prevalence and antibiotic sensitivity of bacteria causing urinary tract infections among non-insulin dependent diabetic patients as well as determining the bacterial causative agents of urinary tract infections through biochemical confirmatory tests. The study was carried out in Kisii Teaching and Referral Hospital in Kisii County, Kenya. One hundred and eighty diabetic patients were enrolled in a cross-sectional study design. Clean catch midstream urine was collected from all participants and cultured in cysteine lactose electrolyte deficient agar for bacteria isolation. The isolates were later cultured in Mueller Hinton for antibiotic sensitivity testing. Classification of a positive culture for urinary tract infection was based on more than 100,000 (≥105) colony-forming units of a single bacterial species. All the data was extracted and analyzed with the statistical package for the social science (SPSS) version 20. The data was analyzed using frequencies, chi square (P<0.05) and logic regression to find the odds ratio. One hundred and seven participants were male (59.4 %) and 73 (40.6 %) were female. Sixty-three participants (35 %) showed symptoms of urinary tract infections. The overall prevalence of urinary tract infections was 20.6 % with 37 participants testing positive for urinary tract infection. The duration of diabetes mellitus was revealed to be significant (p=0.01) while age was also found to have a significant association with urinary tract infection (p=0.002). Out of the 37 (100 %) isolates, 35 (94.6 %) were gram negative and the remaining 2 (5.4 %) were gram positive. Escherichia coli was the most predominant isolate with 21 (56.8 %) isolates followed by 10 (27 %) isolates of Klebsiella pneumonia and then 4 (10.8 %) isolates of Proteus mirabilis. There were two (5.4 %) isolates of Enterococcus faecalis. Out of the 21 E. coli isolates, five isolates showed resistance to ampicillin, three isolates were resistant to nitrofurantoin and three isolates were resistant to co-trimoxazole. Out of 10 K. pneumoniae isolates, two were resistant to ampicillin, one was resistant to cephalexin and two were resistant to co-trimoxazole. Out of the four P. mirabilis isolates, there were three cases where one strain was each resistant to ampicillin, nitrofurantoin and cotrimoxazole. All 21 isolates of E. coli (100 %) were sensitive to gentamicin and cephalexin. All ten K. pneumoniae isolates (100 %) were sensitive to gentamicin and nitrofurantoin. These findings suggest an increasing antibiotic resistance among pathogenic causative agents of UTI among non-insulin dependent diabetic patients. A longitudinal study is recommended with a higher number of participants, in order to understand the risk factors of urinary tract infection among diabetic patients
Evaluation of Enriched Rapid Diagnostic Method for Detection of Vibrio Cholerae and Antibiotic Susceptibility Patterns of Serotypes in Juba, South Sudan
(Kenyatta University, 2018-09) Nyangena, Ontweka Lameck
The global cholera burden is not precisely known since many of the cases lack proper investigation and documentation systems. More importantly, the situation is exacerbated by defective epidemiological surveillance as well as inadequate diagnostic procedures. Isolation of Vibrio cholerae is critical for disease outbreak confirmation. Rapid diagnostic testing of fecal specimens, on basis of lipopolysaccharide detection of V. cholerae O1 or O139 sub-types may enhance early outbreak detection and surveillance. The gold standard for V. cholerae diagnosis is stool culture, although, it’s not feasible in resource constrained settings since it warrants greater than 24 hours of bacterial isolation. As such, the present investigation aimed at evaluating the enriched rapid diagnostic method for V. cholerae detection while also assessing antimicrobial susceptibility patterns of associated V. cholerae serotypes present in Juba, South Sudan. A cross-sectional laboratory-based experimental study was conducted on 109 consenting participants in Juba, South Sudan. Purposive sampling technique was employed. Stool samples were collected and tested using both conventional and the rapid enriched method (alkaline peptone water; APW), with results obtained being compared to V. cholerae stool cultures (gold standard). On the other hand, V. cholerae sero-typing was undertaken using both polyvalent and monovalent antisera. Additionally, antimicrobial susceptibility testing (AST) was carried out on study samples to determine drug sensitivity profiles. All tests were conducted onsite with exception of cultures and AST which were performed in public health laboratory Juba, South Sudan. Out of 109 stool samples only 31.2 % (n=34) were positive for V. cholerae; Inaba serotype 100 % (n=34). Demographic information revealed that 70.6 % (n=24) of all V. cholerae infections were among males aged (26-30 yrs; 32.4 %). Data on antibiotic susceptibility pattern showed that V. cholerae was most sensitive to ceftriaxone (n=31; 91.2 %). Conversely, V. cholerae demonstrated highest resistance against nalidixic acid (n=34; 100 %). Conclusively, enriched method for detection of V. cholerae exhibited notable diagnostic performance compared to V. cholerae culture (control).Sensitivity for enriched and Culture was 68 % in O1 and 0 % in O139 respectively while specificity for enriched was 100 % in O1 and 99.08 % in O139 and on culture was 100 % in both specificity and sensitivity. Additionally, Inaba serotype of V. cholerae showed highest prevalence amongst subjects attending cholera treatment centers and oral rehydration points in Juba, South Sudan. On the other hand, ceftriaxone displayed highest potency agent against V. cholerae. The study concludes that there was high prevalence of Inaba serotype in the region. The findings also inform selection of ceftriaxone as an effective chemotherapeutic agent against V. cholerae Inaba serotype.
Isolation, Antibiotic Susceptibility and Molecular Characterization of Resistance Genes in Pseudomonas Isolates from Selected Hospitals in Mombasa County, Kenya.
(Kenyatta University, 2018-09) Mwinyikombo, Issa Suleiman
Pseudomonas species are common causes of nosocomial infections. Globally nosocomial infections are among the most important problems complicating health care provision since they are caused by bacteria exhibiting resistance to many antibiotics. Their susceptibility to antibiotics is restricted to merely few drugs, and the occurrence of resistance by initially susceptible strains during therapy occurs at a relatively high frequency. The rate of transmission of Pseudomonas infection increases from 5% in non-hospitalized patients to 20% in hospitalized patients within 72 hours after hospital admission. These infections may be contributed by indwelling catheters, surgical or burn wounds as well as abscess in patients who have been admitted for over 48 hours in the hospital. This study was conducted in two selected hospitals in Mombasa County, coastal region of Kenya. A cross sectional study design was adopted and 192 clinical samples from each study hospital were randomly collected and analyzed. Samples were collected from indwelling catheters, swabs from burn wounds, surgical wounds and abscess as well as tubing’s from patients who had stayed for over 48 hours in the hospital wards or ICU. Specimens were inoculated in MacConkey and Blood agar, and later sub cultured in blood agar for purity. Species identification and antimicrobial susceptibility was analyzed by the Vitek® system. Pseudomonas positive isolates were screened for the presence of Metallo-beta lactamase and fluoroquinolones resistance conferring genes by conventional PCR. Out of the 384 samples, collected 50 were positive for Pseudomonas species. The type of specimens collected included 226 (58.9%) pus swabs, 124 (32.3%) catheter tips, 29 (7.6%) endotracheal tube tips and 5 (1.3%) central venous catheter tips. The predominant isolate identified was Pseudomonas aeruginosa 41 (82%), followed by Pseudomonas fluorescence 3 (6%), Pseudomonas stutzeri 3 (6%), Pseudomonas luteola 2 (4%), and Pseudomonas oleovorans 1 (2%). The relative distribution of the Pseudomonas species amongst the two study facilities was not statistically significant (p= 0.955). Pseudomonas isolates were detected in all the specimen types except central venous catheter tips. Pus swabs recorded the highest Pseudomonas burden n=30 (60%) followed by catheter tips n=11 (22%) and finally endotracheal tubes tips n=9 (18%). Antibiotic susceptibility against 26 drugs used was varying. This study observed that (96%) of Pseudomonas isolates were resistant to Ampicillin while Colistin was the most effective antibiotic. Metallo-betalactamase (MBL) and fluoroquinolone resistance conferring genes were analyzed from twenty (20) Pseudomonas isolates using PCR. VIM was the most prevalent MBL gene observed followed by SPM, SIM, IMP, and GIM. Fluoroquinolone resistant genes ParC and MexR were detected in all the isolates screened in this study. This study is of great importance to the county, as it will outline the suitable antibiogram for the treatment of Pseudomonas infections and serves as a baseline to evaluate the trends in antibiotic resistant genes in the region.

Browse

Browsing MST-Department of Medical Laboratory Sciences by Issue Date

Results Per Page

Sort Options