Development, Optimization and Evaluation of a Sensitive Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detection of Chicken-Based IgY Polyclonal Antibodies against Toxins of D. polylepis Venom.

Life-threatening medical issues can result from snakebite, and hence this is a public health concern. In many tropical and subtropical nations such as Kenya, where a wide variety of poisonous snakes are prevalent, diagnosis of snakebite in health facilities is imperative. Different antivenoms are needed to treat the venom of different snake species. Nonetheless, it might be difficult for medical professionals to identify the exact snake species that envenomated a patient due to the similarities of several snake envenomations' clinical symptoms. Therefore, the necessity for an assay or technique for identifying venomous species is critical. The current study sought to develop a sensitive ELISA prototype for the detection of D. polylepis venom in Kenya using generated chicken-based IgY polyclonal antibodies. Serum samples containing specific chicken-based IgY antibodies previously raised against D. polylepis venom toxins were used in the assay development. ELISA parameters were optimized, and the developed assay was assessed for applicability. The limit of detection (LoD) of the ELISA for neurotoxic venoms was determined to be 0.01 µg/mL. Successful discrimination between neurotoxic and cytotoxic venoms was achieved by the ensuing inhibition ELISA assay. The developed assay showed the capability of identifying venoms in blood samples (from spiked and venom-challenged blood samples) of BALB/c mice, providing compelling evidence of the strategy's usefulness. This assay could help physicians diagnose and manage victims of snakebites through the evaluation of clinical samples.

Generation of chicken-based IgY polyclonal antibodies against Dendroaspis polylepis and preclinical evaluation of envenomation-neutralizing efficacy vis-à-vis selected commercial antivenoms.

The Black mamba, D. polylepis, is one of the many venomous snakes found in Kenya, and known to account for some snakebite incidents. The Kenyan Ministry of Health data reveals annual 15,000 snakebites occurrences. Also, 1 in 15 people in Kenya gets bitten by a snake, and tragically, 1 in 147 of these individuals die of snakebite yearly. Traditionally, antivenoms for treatment are produced from horse or sheep but have complicated and expensive production issues. Alternative production approaches, such as using IgY antibodies derived from chicken egg yolks, may overcome disadvantages with traditional antivenom manufacturing techniques. In this current study, D. polylepis specific IgY polyclonal antibodies were purified from the egg yolks of chickens immunized with D. polylepis venom. These antibodies were subsequently assessed for their in-vivo neutralizing capacity vis-à-vis commercial antivenoms, PANAF-Premium and VINS. The IgY antibodies were purified by ammonium sulfate precipitation and affinity-chromatography, with quality and specificity determined by SDS-PAGE and ELISA. The LD50 of D. polylepis was found to be 0.54 mg/kg in chicks, and 0.34 mg/kg in mice, respectively. Pool of extracted IgY yielded 2.8 mg/mL concentration. Purified IgY under non-reducing and reducing conditions on SDS-PAGE exhibited a single-protein band of about 183 kDa and two bands (67 kDa and 25 kDa), respectively. The minimum-edematogenic dose was 0.05 µg. Anti-D. polylepis IgY antibodies and two antivenoms demonstrated the capacity to neutralize the toxic activities of D. polylepis venom. This study confirms a successful IgY generation against Black mamba venom for the first time, and observed toxic effects of the venom as well as neutralizing capacity of antivenoms.

Comparative prevalence of diarrheagenic Escherichia coli between children below five years with close contact to food animals in Kisumu County, Kenya.

Introduction: diarrheal infections in young children below five years and food animals are caused by diarrheagenic Escherichia coli strains. The study focused on understanding the association between DEC pathotypes in children below five years and food animals to establish the possibility of zoonotic transmission. Methods: samples from 150 children who presented with diarrhea at the Kisumu County Hospital and 100 stool samples from food animals were collected and processed using culture methods. Molecular identification of the pathotypes was assayed using a primer-specific polymerase chain reaction that targeted the six virulence genes related to the diarrheagenic Escherichia coli pathotypes. Results: one hundred and fifty-six study subjects (100 children samples and 56 food animals) samples were positive for E. coli polymerase chain reaction detection revealed a prevalence of (23%) among children below five years and a prevalence of (20%) among the food animals. Children samples showed Enteroaggregative Escherichia coli, having high phenotypic frequency of (12%) followed by Enterotoxigenic Escherichia coli, (5.3%) and Enteropathogenic Escherichia (3.3%) the least being mixed infections Enteroaggregative/Enterotoxigenic Escherichia coli and Enteroaggregative/Enteropathogenic Escherichia coli with (1.3%) respectively. The food animals found in children homesteads were detected to harbor pathogenic strains of E. coli. Enteropathogenic Escherichia coli was the most prevalent pathotypes detected in cattle (13%) followed by Enterotoxigenic Escherichia coli detected in goats at (4%) and poultry at (3%). Conclusion: presence of diarrheagenic Escherichia coli in food animals could serve as reservoirs of transmitting these bacteria to children below five years.

Perceptions of HIV infected patients on the use of cell phone as a tool to support their antiretroviral adherence; a cross-sectional study in a large referral hospital in Kenya.

BACKGROUND: Clinical trials were conducted to assess the feasibility of using a cell phone text messaging-based system to follow up Human Immunodeficiency Virus (HIV) infected patients on antiretroviral (ARTs) and assess for improved adherence to their medication. However there is need to evaluate the perceptions of the HIV infected patients towards the use of these cell phones in an effort to better aid in the clinical management of their HIV infection. The objective of this study was therefore to determine the perceptions of HIV infected patients on the use of cell phone text messaging as a tool to support adherence to their ART medication. METHODS: A cross sectional survey was conducted among patients receiving Highly Active Anti-Retroviral Therapy (HAART) at the Kenyatta National Hospital Comprehensive Care Clinic in Nairobi between May and July, 2011. Pre-tested questionnaires were used to collect the socio-demographic and perceptions data. The recruitment of the participants was done using the random probability sampling method and statistical analysis of data performed using Statistical Package for Social Sciences (SPSS) version 16.0. RESULTS: A total of 500 HIV infected patients (Male-107, Female-307) aged 19-72 years were interviewed. The majority of individuals (99%) had access to cell phones and 99% of the HIV infected patients interviewed supported the idea of cell phone use in management of their HIV infection. A large proportion (46%) claimed that they needed cell phone access for medical advice and guidance on factors that hinder their adherence to medication and only 3% of them needed it as a reminder to take their drugs. The majority (72%) preferred calling the healthcare provider with their own phones for convenience and confidential purposes with only 0.4% preferring to be called or texted by the health care provider. Most (94%), especially the older patients, had no problem with their confidentiality being infringed in the process of the conversation as per the bivariate analysis results. CONCLUSION: Cell phone communications are acceptable and in fact preferable over cell phone reminders.

Prognostic factors influencing HIV-free survival among infants enrolled for HIV early infant diagnosis services in selected hospitals in Nairobi County, Kenya.

BACKGROUND: Despite being a preventable disease, pediatric HIV infection continues to be a public health concern due to the morbidity and mortality associated with the disease. Vertical transmission of HIV occurs when a mother living with HIV passes the virus to her baby during pregnancy, childbirth, or breastfeeding. Globally, the vertical transmission rate of HIV is 9% with sub-Saharan Africa accounting for 90% of these infections. In Kenya, the national vertical transmission rates of HIV stood at 11.5% by the end of 2018, with a target to reduce vertical transmission rates to below 5% and 2% in breastfeeding and non-breastfeeding infants respectively, by the end of 2021. OBJECTIVE: To determine the prognostic factors influencing HIV-free survival among infants enrolled for HIV early infant diagnosis (EID) services in selected hospitals in Nairobi County, Kenya. METHODS: A prospective cohort study design was adopted. HIV exposed infants were recruited at six weeks to determine HIV-free survival over 12 months follow up. Simple random sampling was used to select 166 infants and data were collected from the mothers using semi-structured interviewer-administered questionnaires. Log-rank tests were used to test for associations at the bi-variable level while Cox-proportional regression was used to analyze data at the multi-variable level, with the aid of STATA 14 software. Ethical approval was obtained from Kenya Medical Research Institute, Scientific Ethics Review Unit. RESULTS: The overall infant HIV incidence rate over one-year follow-up was 9 cases per 100 person-years (95% CI: 5.465-16.290). The failure event was defined as an infant with a positive PCR test during the study period with total failures being 13 (9.41%) over 12 months. Prognostic factors associated with poor infant HIV-free survival were young maternal age (18-24 years) and mothers with a recent HIV diagnosis of ≤ 2 years since a positive HIV diagnosis (HR 5.97 CI: 1.20, 29.58) and (HR 6.97 CI: 1.96, 24.76), respectively. CONCLUSION: Maternal prognostic factors associated with poor infant HIV-free survival were young maternal age (18-24 years) and recent maternal HIV diagnosis of ≤ 2 years since positive HIV diagnosis. The study recommended the development of an intervention package with more rigorous adherence counseling and close monitoring for young mothers, and mothers with recent HIV diagnoses.

Utilization of digital tools to enhance COVID-19 and tuberculosis testing and linkage to care: A cross-sectional evaluation study among Bodaboda motorbike riders in the Nairobi Metropolis, Kenya.

Kenya has registered over 300,000 cases of COVID-19 and is a high-burden tuberculosis country. Tuberculosis diagnosis was significantly disrupted by the pandemic. Access to timely diagnosis, which is key to effective management of tuberculosis and COVID-19, can be expanded and made more efficient through integrated screening. Decentralized testing at community level further increases access, especially for underserved populations, and requires robust systems for data and process management. This study delivered integrated COVID-19 and tuberculosis testing to commercial motorbike (Bodaboda) riders, a population at increased risk of both diseases with limited access to services, in four counties: Nairobi, Kiambu, Machakos and Kajiado. Testing sheds were established where riders congregate, with demand creation carried out by the Bodaboda association. Integrated symptom screening for tuberculosis and COVID-19 was conducted through a digital questionnaire which automatically flagged participants who should be tested for either, or both, diseases. Rapid antigen-detecting tests (Ag-RDTs) for COVID-19 were conducted onsite, while sputum samples were collected and transported to laboratories for tuberculosis diagnosis. End-to-end patient data were captured using digital tools. 5663 participants enrolled in the study, 4946 of whom were tested for COVID-19. Ag-RDT positivity rate was 1% but fluctuated widely across counties in line with broader regional trends. Among a subset tested by PCR, positivity was greater in individuals flagged as high risk by the digital tool (8% compared with 4% overall). Of 355 participants tested for tuberculosis, 7 were positive, with the resulting prevalence rate higher than the national average. Over 40% of riders had elevated blood pressure or abnormal sugar levels. The digital tool successfully captured complete end-to-end data for 95% of all participants. This study revealed high rates of undetected disease among Bodaboda riders and demonstrated that integrated diagnosis can be delivered effectively in communities, with the support of digital tools, to maximize access.

Determinants of microbial contamination of cooked foods hawked in Tharaka Nithi County, Kenya: a cross-sectional study.

Introduction: hawking of cooked foods is an important socio-economic activity. However, hawked cooked foods are prone to contamination, especially by pathogenic microorganisms. Methods: this was a cross-sectional study of 151 cooked food hawkers and 151 cooked food samples, one sample each from every respondent. Data was collected through questionnaires, observation checklists, laboratory processes, and key informant interviews. Quantitative data were analyzed using SPSS, while qualitative data were analyzed using the thematic network analysis technique. Results: adjusted multinomial logistic regression revealed increased odds of microbial contamination in cooked foods hawked by persons with primary-level education (adjusted odds ratio) AOR = 9.278, 95% confidence interval (CI) 2.292, 37.548; p = 0.003), persons earning a profit of Ksh. ≤500.00 (AOR = 4.046, 95% CI 1.825, 8.969; p = 0.001), and in areas infested with rodents (AOR = 6.386, 95% CI 1.877, 21.724; p = 0.003). However, cooked foods hawked by males (AOR = 0.338, 95% CI 0.133, 0.856; p = 0.022), persons trained on food safety (AOR = 0.216, 95% CI 0.064, 0.733; p = 0.014), persons wearing outer garments (AOR = 0.399, 95% CI 0.196, 0.812; p = 0.011), and by persons who stored their garbage in a municipal receptacle or a standard dust bin (AOR = 0.13, 95% CI 0.041, 0.410; p = 0.000) recorded reduced risk of contamination. Conclusion: hawker´s characteristics, food handling practices, and the condition of the hawking station are important determinants of microbial contamination of cooked foods hawked in Tharaka Nithi County. Targeted interventions need to be developed and implemented to prevent related foodborne diseases.

Neisseria gonorrhoeae infection in female sex workers in an STI clinic in Nairobi, Kenya.

BACKGROUND: Gonorrhea caused by Neisseria gonorrhoeae is the second most prevalent curable sexually transmitted infection worldwide. Female Sex Workers (FSWs) are at a higher risk of contracting gonorrhea due to their risky sexual behaviors like inconsistent condom use and multiple sexual partners. We determined the prevalence and risk factors associated with gonorrhea and its antimicrobial susceptibility pattern among symptomatic FSWs attending Sexual Workers Outreach Program (SWOP) city clinic in Nairobi, Kenya. METHODS: Using convenience sampling, we recruited 379 female sex workers from SWOP City clinic in Nairobi County. We administered a semi-structured questionnaire to collect data on socio-demographics and behavioral risk factors associated with gonorrhea. We also conducted three focus groups. Two endocervical swabs were collected from each participant by the attending physician for the laboratory identification of Neisseria gonorrhoeae. An antimicrobial susceptibility test was performed using the disc diffusion method. RESULTS: Twenty-four out of 379 (6.3%) participants tested positive for gonorrhea by PCR. The significant risk factors associated with gonorrhea were having multiple sexual partners in the previous two weeks, primary education, and being in the age group of 38-49 years (p < 0.05). From the qualitative data, sex work disclosure, and difficulty in engaging protected sex with their partner, and unprotected sex with their clients due to more money from the client, PREP, and alcohol use made the female sex workers vulnerable to gonorrhea exposure and or risky sexual behavior. The culture-positive sample result yielded complete (100%) resistance to all the antimicrobials used. CONCLUSION: Neisseria gonorrhoeae infection is prevalent among symptomatic FSWs in Nairobi. Multiple sexual partners, being in age group 38-49 years and having primary education were the factors associated with gonorrhea among the study participants. Based on our identification of a highly resistant isolate, we strongly recommend increasing capacity for culture-based diagnosis and susceptibility testing.

Factors associated with the risk of suicidal behavior among adolescents transitioning to secondary school in Nairobi County, Kenya: a cross-sectional study.

Introduction: adolescence is a transitory stage in the lives of young people. The transition from primary to secondary school among adolescents is associated with suicidal behavior but is not well characterized in the Kenyan context. This study sought to elucidate factors associated with the risk of suicidal behavior among adolescents aged 11-18 years in transition to secondary school. Methods: a cross-sectional design was employed in the study that was conducted among adolescents in 5 randomly selected secondary schools in Nairobi County. The study involved 539 students who had joined form 1 in January 2020. Data were collected using the suicide behavior questionnaire-revised (SBQ-R) in March 2020. Factors associated with suicidal behavior were assessed using a generalized linear model (GLM), using a poisson distribution with a log-link function to estimate adjusted prevalence ratios (aPR), and a significance level of p=.05. Results: one-fifth (20.04%) of adolescents with a median age of 14 years were at risk of suicidal behavior. Depression (aPR=3.16, C.I {1.85, 5.41}, p=0.001) and lifetime alcohol use (aPR=1.87, C.I {1.17, 2.97}, p=0.009) were found to be significant factors for suicidal behavior. Conclusion: depression and lifetime alcohol use are associated with the risk of suicidal behavior among adolescents transitioning from primary to secondary school. Interventions may need to be targeted at the pre-secondary or primary school level to prevent underage alcohol use and enhancement of social support to prevent depression in this demographic of the population.

Effects of bacterial vaginosis-associated bacteria and sexual intercourse on vaginal colonization with the probiotic Lactobacillus crispatus CTV-05.

OBJECTIVE: Several fastidious bacteria have been associated with bacterial vaginosis (BV), but their role in lactobacilli recolonization failure is unknown. We studied the effect of 7 BV-associated bacterial species and 2 Lactobacillus species on vaginal colonization with Lactobacillus crispatus CTV-05 (LACTIN-V). METHODS: Twenty-four women with BV were given a 5-day course of metronidazole vaginal gel and then randomized 3:1 to receive either LACTIN-V or placebo applied vaginally once daily for 5 initial consecutive days, followed by a weekly application over 2 weeks. Vaginal swabs for L. crispatus CTV-05 culture and 9 bacterium-specific 16S rRNA gene quantitative polymerase chain reaction assays were analyzed on several study visits for the 18 women receiving LACTIN-V. RESULTS: Vaginal colonization with CTV-05 was achieved in 61% of the participants receiving LACTIN-V at either day 10 or day 28 visit and 44% at day 28. Participants not colonized with CTV-05 had generally higher median concentrations of BV-associated bacteria compared to those who colonized. Between enrollment and day 28, the median concentration of Gardnerella vaginalis minimally reduced from 10 to 10 16S rRNA gene copies per swab in women who colonized with CTV-05 but increased from 10 to 10 in those who failed to colonize (P = 0.19). Similarly, the median concentration of Atopobium spp. reduced from 10 16S rRNA gene copies per swab to below limit of detection in women who colonized with CTV-05, but increased from 10 to 10 in those who failed to colonize (P = 0.04). The presence of endogenous L. crispatus at enrollment was found to be significantly associated with a reduced odds of colonization with CTV-05 on day 28 (P = 0.003), and vaginal intercourse during the study significantly impaired successful CTV-05 colonization (P = 0.018). CONCLUSION: Vaginal concentration of certain BV-associated bacteria, vaginal intercourse during treatment, and the presence of endogenous L. crispatus at enrollment predict colonization with probiotic lactobacilli.

Influence of hepatitis B virus co-infection on virological and immunological response to antiretroviral treatment among HIV patients attending comprehensive care clinics in Makueni County, Kenya.

INTRODUCTION: the effect of hepatitis B virus (HBV) infection on the natural history of human immunodeficiency virus (HIV) disease remains uncertain. Therefore, this study was conducted to determine the association of HBV co-infections with CD4 count and viral load levels in response to antiretroviral treatment among HIV patients attending comprehensive care clinics in Makueni County (Kenya). METHODS: this was a prospective case-control study among patients seeking HIV services in three hospitals of Makueni County (Kenya). Newly diagnosed HIV mono-infected patients (controls) and HIV/HBV co-infected (cases), 18 years and above who had not started antiretrovirals (ARVs) participated. A total of 258 patients gave informed consent and participated. HIV mono-infected (controls) produced 129 while HIV/HBV (cases) gave 129 participants. P-values ≤ 0.05 were considered significant. RESULTS: the majority (164, 63%) of the study participants were females for both arms of the study. The mean age of the participants was 31±0.402 years and majority of them were between the age of 26-30years old. At the beginning and end of the study the mean viral load for HIV/HBV co-infected individuals was (30169 and 1731) copies/ml while that of CD4 count was (327 and 459) cells/ul, and that of HIV mono-infected was (21860 and 1689) copies/ml and CD4 count of (421 and 437) cells/ul respectively. After enrolling them into antiretroviral therapy (ART) treatment and after six months of follow-up there was significant drop in viral load and significant increase in CD4 count for both groups at p<0.001 using logistic regression. CONCLUSION: HIV patients co-infected with hepatitis B virus had high viral load and low CD4 count compared to HIV monoinfected patients however with introduction of ARVs there was improvement in both groups with the highest noticed among the HIV/HBV co-infected patients.

Risk factors for hypertension among young adults (18-35) years attending in Tenwek Mission Hospital, Bomet County, Kenya in 2018.

INTRODUCTION: Hypertension ranks third in the world, after underweight and unsafe sex, in the list of six major risk factors contributing to the global disease. In Kenya, the prevalence stands at 24% in the general population, while among the young adults, the incidence of hypertension has been reported to be in the rise; a fact attributed to increased number of risks. We therefore sought to determine awareness and risk factors of hypertension among young adults attending Tenwek hospital. METHODS: A case-control study of young adults ages 18-35, involving 80 cases and 80 controls at Tenwek Mission Hospital, Bomet County. Cases included males and females newly diagnosed with hypertension (diagnosed at the time of data collection) and if they reported taking antihypertensive medication and reported as hypertensives in the hospital records at any clinic visit or at interview, while controls included persons with no history of hypertension. RESULTS: Those having a BMI≥25 were 3.05 times more likely to be hypertensive (OR: 3.05, 95% CI 1.26, 7.40; p=0.014). Having a relative suffering from hypertension increased almost thrice the odds of being hypertensive (OR: 2.78, 95% CI 1.20, 6. 46; p=0.018). Not drinking alcohol reduced the chance of suffering from hypertension by 70%, (OR=0.30, 95% CI 0.11, 0.81; p=0.017). CONCLUSION: The prevalence of hypertension in younger adults is not as low as generally perceived. Preventive measures should be formulated in a manner to address variety of major risk factors in young adults.

Prevalence, aetiology and antibiotic sensitivity profile of asymptomatic bacteriuria isolates from pregnant women in selected antenatal clinic from Nairobi, Kenya.

INTRODUCTION: Asymptomatic bacteriuria (ASB) is the presence of bacteria in urine without apparent symptoms of urinary tract infections. The importance of asymptomatic bacteriuria lies in the insight it provides into symptomatic infections. To determine prevalence, bacterial isolates and Antibiotic Sensitivity Profile of asymptomatic bacterial urinary tract infection in pregnant women in selected clinics in Nairobi. METHODS: This was a cross-sectional study involving women attending antenatal clinic at selected clinics of Nairobi County. The women who met the inclusion criteria were included in the study. The midstream urine samples of these women were subjected to microscopy, culture and sensitivity. RESULTS: A total of 1020 of women on their first antenatal clinic visit participated in the study; 219 of them had ASB, giving a prevalence of 21.5 % at 95% confidence level. Escherichia coli were the common organism isolated at 38.8%. The majority of the organisms were sensitive to imipenem and gentamycin. CONCLUSION: There is a high prevalence of ASB among pregnant women included in the study from the Nairobi county clinics. Therefore, routine ASB screening of pregnant women is recommended among the women attending antennal clinics in Nairobi county clinics.

Factors influencing knowledge on completion of treatment among TB patients under directly observed treatment strategy, in selected health facilities in Embu County, Kenya.

INTRODUCTION: The World Health Organization (WHO) promotes the Directly Observed Treatment (DOT) strategy as the standard to increase adherence to Tuberculosis (TB) medication. However, cases of retreatment and Multi Drug Resistant continue to be reported in many parts of Kenya. This study sought to determine the factors influencing the completion of tuberculosis medication among TB patients in Embu County, Kenya. METHODS: A descriptive cross-sectional study was conducted on a population of tuberculosis patients under DOT attending selected TB treatment clinics in Embu County, in Kenya. One hundred and forty TB patients interviewed within a period of 3 months. Data were analyzed using SPSS version 17.0 and included Bivariate and Multivariate Analysis. The level of significance was p≤ 0.05. RESULTS: The male and female participants were 61.4% and 38.6% respectively. The mean age of the respondents was 35±31.34-39.3 years. For the majority (52%) of the participants, the highest level of education was primary education. The unemployed participants formed the highest number of the respondent in the study (73%). The majorities (91.4%0) of the respondents were under the home-based DOT strategy (91.4%, 95% C.I: 85.5-95.5). Bivariate analysis using Chi-square showed that the level of education (p=0.003), patients feeling uncomfortable during supervision (p=0.01), and knowledge regarding the frequency of taking medication (p=0.004) were all significantly associated with knowledge regarding the importance of completion of medication. However, none of these factors was significant after multivariate analysis. CONCLUSION: Most participants did not know the importance of completion of medication. TB programs should come up with better ways to educate TB patients on the importance of supervision and treatment completion during the treatment of TB. The education programs should focus on influencing the attitudes of patients and creating awareness about the importance of treatment completion. The TB programs should be designed towards eliminating the factors influencing the completion of TB medication.

High Prevalence of HIV Low Abundance Drug-Resistant Variants in a Treatment-Naive Population in North Rift Kenya.

The advent of antiretroviral treatment (ART) has resulted in a dramatic reduction in AIDS-related morbidity and mortality. However, the emergence and spread of antiretroviral drug resistance (DR) threaten to negatively impact treatment regimens and compromise efforts to control the epidemic. It is recommended that surveillance of drug resistance occur in conjunction with scale-up efforts to ensure that appropriate first-line therapy is offered relative to the resistance that exists. However, standard resistance testing methods used in Sub-Saharan Africa rely on techniques that do not include low abundance DR variants (LADRVs) that have been documented to contribute to treatment failure. The use of next generation sequencing (NGS) has been shown to be more sensitive to LADRVS. We have carried out a preliminary investigation using NGS to determine the prevalence of LDRVS among a drug-naive population in North Rift Kenya. Antiretroviral-naive patients attending a care clinic in North Rift Kenya were requested to provide and with consent provided blood samples for DR analysis. DNA was extracted and amplified and nested PCR was conducted on the pol RT region using primers tagged with multiplex identifiers (MID). Resulting PCR amplicons were purified, quantified, and pyrosequenced using a GS FLX Titanium PicoTiterPlate (Roche). Valid pyrosequencing reads were aligned with HXB-2 and the frequency and distribution of nucleotide and amino acid changes were determined using an in-house Perl script. DR mutations were identified using the IAS-USA HIV DR mutation database. Sixty samples were successfully sequenced of which 26 were subtype A, 9 were subtype D, 2 were subtype C, and the remaining were recombinants. Forty-six (76.6%) had at least one drug resistance mutation, with 25 (41.6%) indicated as major and the remaining 21 (35%) indicated as minor. The most prevalent mutation was NRTI position K219Q/R (11/46, 24%) followed by NRTI M184V (5/46, 11%) and NNRTI K103N (4/46, 9%). Our use of NGS technology revealed a high prevalence of LADRVs among drug-naive populations in Kenya, a region with predominantly non-B subtypes. The impact of these mutations on the clinical outcome of ART can be ascertained only through long-term follow-up.

Knowledge and practices towards rabies and determinants of dog rabies vaccination in households: a cross sectional study in an area with high dog bite incidents in Kakamega County, Kenya, 2013.

INTRODUCTION: An estimated 55,000 people die from rabies annually. Factors promoting dog vaccination, estimates of vaccination coverage and knowledge on rabies are important for effective rabies control. We sought to establish these estimates at household (HH) level and whether rabies knowledge is associated with proper control practices. METHODS: Cross-sectional cluster survey with two-stage sampling was employed in Kakamega County to enroll HH members above 18 years. A set of questions related to rabies knowledge and practice were used to score participant response. Score above the sample mean was equated to adequate knowledge and proper practices respectively. Independent t-test was used to evaluate the differences of sample mean scores based on dog vaccination status. Bivariate analysis was used to associate knowledge to practices. RESULTS: Three hundred and ninety HHs enrolled and had a population of 754 dogs with 35% (n = 119) HH having vaccinated dogs within past 12 months. Overall mean score for knowledge was 7.0 (±2.8) with range (0-11) and 6.3 (±1.2) for practice with range (0-8). There was a statistically significant difference in mean knowledge (DF = 288, p < 0.01) and practice (DF = 283, p = 0.001) of HH with vaccinated dogs compared to ones with unvaccinated dogs. Participants with adequate rabies knowledge were more likely to have proper health seeking practices 139 (80%) (OR = 3.0, 95% CI = 1.4-6.8) and proper handling practices of suspected rabid dog 327 (88%) (OR = 5.4, 95% CI = 2.7-10.6). CONCLUSION: Rabies vaccination below the 80% recommended for herd immunity. Mass vaccination campaign needed. More innovative ways of translating knowledge into proper rabies control practice are warranted.

A cross-sectional study on the microbiological quality and safety of raw chicken meats sold in Nairobi, Kenya.

BACKGROUND: Chicken is a rich source of meat protein and is increasingly being consumed in urban areas in Kenya. However, under poor hygienic environment, raw chicken meat presents an ideal substrate supporting the growth of pathogenic Escherichia coli and Coliform bacteria indicating the potential presence of other pathogenic bacteria; this may constitute a major source of food-borne illnesses in humans. This study sought to assess the microbiological quality and safety of raw chicken meat sold in Nairobi, Kenya by determining the E. coli/coliform contamination levels as well as the antimicrobial resistance patterns and pathogenicity of E. coli isolated. FINDINGS: We conducted a Cross-sectional study to collect two hundred raw chicken samples that were randomly purchased between the periods of August 2011-February 2012. Enumeration of bacteria was done using 3 M Petri film E. coli/Coliform count plates, isolation and identification of E. coli through standard cultural and biochemical testing, antimicrobial susceptibilities interpreted according to criteria set by the Clinical and Laboratory Standards Institute (2012) while Polymerase chain reaction assays were used to determine presence of virulence genes in isolated E. coli. Data was analyzed using SPSS version 17.0. Contamination rates were 97% and 78% respectively for Coliform bacteria and E. coli. Seventy six percent of samples fell under the unacceptable microbial count limit (>100 cfu/ml) and significant differences in the E. coli/coliform counts (p < 0.001) were observed among the chicken retail outlets with samples from supermarkets having the lowest level of contamination compared to the rest of the retail outlets. Seventy five percent of the isolates were resistant to at least one of the 12 antibiotics tested with resistance to tetracycline being the highest at 60.3%. In addition 40.4% E. coli isolates were positive for the ten virulence genes tested. CONCLUSION: Raw retail chicken meats in Nairobi are not only highly contaminated, but also with potentially pathogenic and multi-drug resistant strains of E. coli. It will be important for public health authorities and retail chicken processing outlets to collaborate in ensuring adherence to set out principles of hygienic processing and handling of chicken meats in order to reduce potential risks of infection.

Aetiology and factors associated with bacterial diarrhoeal diseases amongst urban refugee children in Eastleigh, Kenya: A case control study.

INTRODUCTION: Kenya is home to over 400 000 refugees from neighbouring countries. There is scanty information about diarrhoea amongst urban refugees in Kenya. OBJECTIVES: We investigated the enteric bacteria causing diarrhoea amongst urban refugee children and described the associated factors. METHOD: During the period of August-December 2010, urban refugee children between the ages of two and five who attended Eastleigh County Council Health Centre were enrolled into the study. Diarrhoeal cases were compared with age-matched children with no diarrhoea (controls). Stool specimens were collected and enteric bacteria isolated. A questionnaire was administered to identify risk factors. RESULTS: A total of 41 cases and 41 controls were enrolled in the study. The age and country of origin were similar for cases and controls. The bacterial isolation rates amongst the cases were: non-pathogenic Escherichia coli 71%, Shigella dysenteriae 2.4%, Shigella flexneri 2.4%, Salmonella paratyphi 5%. For the controls, non-pathogenic E. coli 90% and enterotoxigenic E. coli (ETEC) 2.4% were amongst the organisms isolated. All isolates were resistant to amoxicillin; resistance to other antibiotics varied by isolate type. Factors associated independently with diarrhoea included children not washing their hands with soap (aOR 5.9, p < 0.05), neighbour(s) having diarrhoea (aOR 39.8, p < 0.05), children not exclusively breastfed for their first 6 months (aOR 7.6, p < 0.05) and children eating food cooked the previous day (aOR 23.8, p = 0.002). CONCLUSIONS: Shigella species, Salmonella species and ETEC were found to be responsible for diarrhoea amongst the urban refugee children. Measures to control and guide the use of antibiotics are critical for the prevention of antibiotic resistance. Efforts to improve personal and domestic hygiene, including educational campaigns to promote appropriate handwashing, should be encouraged.

Risk factors of severe pneumonia among children aged 2-59 months in western Kenya: a case control study.

INTRODUCTION: Globally, pneumonia is the leading cause of death in children under the age of 5 years. In Kenya, it is the second leading cause of mortality, accounting for greater than 30,000 deaths in this age group annually. This study sought to identify risk factors for severe pneumonia in children under the age of five years. METHODS: We conducted a case control study. Cases were children aged 2 to 59 months with severe pneumonia or very severe pneumonia and controls were those with non-severe pneumonia as defined by the integrated management of childhood illnesses classification. We administered structured questionnaires to mothers of participants to obtain data on socio-demographics, nutritional status and potential environmental risk factors. Data was analyzed using Epi Info; significance level was set at 0.05. RESULTS: We recruited 103 cases and 103 controls. The median age of cases was 14.0 (Range 3-58) months and of controls 14.0 (Range 2-54) months. Comorbidity (Odds Ratio = 3.8, Confidence Interval 1.4-10.6), delay in seeking treatment for three days or more (Odds Ratio = 2.3, Confidence Interval 1.2-4.2) and contact with upper respiratory tract infection (Odds Ratio = 2.7, Confidence Interval 1.1-6.5) were independent risk factors for severe pneumonia. Receiving antibiotics at home (Odds Ratio = 0.4, Confidence Interval 0.2-0.8) was protective. CONCLUSION: Co-morbidity, contact with upper respiratory tract infection and delay in seeking treatment are risk factors for severe pneumonia. We recommend health education regarding appropriate health seeking and engaging community health workers in pneumonia prevention, control and treatment.