Analysis of Cellular Stress Assay Parameters and Intracellular ATP in Platelets: Comparison of Platelet Preparation Methods.

Platelets are metabolically active, anucleated and small circulating cells mainly responsible for the prevention of bleeding and maintenance of hemostasis. Previous studies showed that platelets mitochondrial content, function, and energy supply change during several diseases such as HIV/AIDS, COVID-19, pulmonary arterial hypertension, and in preeclampsia during pregnancy. These changes in platelets contributed to the severity of diseases and mortality. In our previous studies, we have shown that the seahorse-based cellular stress assay (CSA) parameters are crucial to the understanding of the mitochondrial performance in peripheral blood mononuclear cells (PBMCS). Moreover, the results of CSA parameters were significantly influenced by the PBMC preparation methods. In this study, we assessed the correlation of CSA parameters and intracellular ATP content in platelets and evaluated the effects of platelet preparation methods on the results of CSA parameters and intracellular ATP content. We compared the results of CSA parameters and intracellular ATP content in platelets isolated by density centrifugation with Optiprep and simple centrifugation of blood samples without Optiprep. Platelets isolated by centrifugation with Optiprep showed a higher spare capacity, basal respiration, and maximal respiration than those isolated without Optiprep. There was a clear correlation between basal respiration and maximal respiration, and the whole-ATP content in both isolation methods. Moreover, a positive correlation was observed between the relative spare capacity and whole-cell ATP content. In conclusion, the results of seahorse-based CSA parameters and intracellular ATP content in platelets are markedly influenced by the platelet isolation methods employed. The results of basal respiration and maximal respiration are hallmarks of cellular activity in platelets, and whole-cell ATP content is a potential hint for basic platelet viability. We recommend further studies to evaluate the role of CSA parameters and intracellular ATP content in platelets as biomarkers for the diagnosis and prediction of disease states.

Molecular epidemiology and transmission dynamics of multi-drug resistant tuberculosis strains using whole genome sequencing in the Amhara region, Ethiopia.

BACKGROUND: Drug resistant Mycobacterium tuberculosis prevention and care is a major challenge in Ethiopia. The World health organization has designated Ethiopia as one of the 30 high burden multi-drug resistant tuberculosis (MDR-TB) countries. There is limited information regarding genetic diversity and transmission dynamics of MDR-TB in Ethiopia. OBJECTIVE: To investigate the molecular epidemiology and transmission dynamics of MDR-TB strains using whole genome sequence (WGS) in the Amhara region. METHODS: Forty-five MDR-TB clinical isolates from Amhara region were collected between 2016 and 2018, and characterized using WGS and 24-loci Mycobacterium Interspersed Repetitive Units Variable Number of Tandem Repeats (MIRU-VNTR) typing. Clusters were defined based on the maximum distance of 12 single nucleotide polymorphisms (SNPs) or alleles as the upper threshold of genomic relatedness. Five or less SNPs or alleles distance or identical 24-loci VNTR typing is denoted as surrogate marker for recent transmission. RESULTS: Forty-one of the 45 isolates were analyzed by WGS and 44% (18/41) of the isolates were distributed into 4 clusters. Of the 41 MDR-TB isolates, 58.5% were classified as lineage 4, 36.5% lineage 3 and 5% lineage 1. Overall, TUR genotype (54%) was the predominant in MDR-TB strains. 41% (17/41) of the isolates were clustered into four WGS groups and the remaining isolates were unique strains. The predominant cluster (Cluster 1) was composed of nine isolates belonging to lineage 4 and of these, four isolates were in the recent transmission links. CONCLUSIONS: Majority of MDR-TB strain cluster and predominance of TUR lineage in the Amhara region give rise to concerns for possible ongoing transmission. Efforts to strengthen TB laboratory to advance diagnosis, intensified active case finding, and expanded contact tracing activities are needed in order to improve rapid diagnosis and initiate early treatment. This would lead to the interruption of the transmission chain and stop the spread of MDR-TB in the Amhara region.

The Determination of Mitochondrial Mass Is a Prerequisite for Accurate Assessment of Peripheral Blood Mononuclear Cells' Oxidative Metabolism.

Mitochondria are responsible for ATP synthesis through oxidative phosphorylation in cells. However, there are limited data on the influence of mitochondrial mass (MM) in the adequate assessment of cellular stress assay (CSA) results in human peripheral blood mononuclear cells (PBMCs). Therefore, the aim of this study was to determine MM in PBMCS and assess its influence on the results of CSA measurements. Blood samples were collected and sent to the laboratory for MM and CSA measurements during different seasons of the year. The mitochondrial mass was determined based on the mtDNA:nDNA ratio in PBMCs using quantitative real-time PCR (qRT-PCR). CSA was measured using Seahorse technology. The MM was significantly lower during summer and autumn compared to winter and spring (p < 0.0001). On the contrary, we found that the maximal respiration per mitochondrion (MP) was significantly higher in summer and autumn compared to winter and spring (p < 0.0001). The estimated effect of MM on mitochondrial performance was -0.002 pmol/min/mitochondrion (p < 0.0001) and a correlation coefficient (r) of -0.612. Similarly, MM was negatively correlated with maximal respiration (r = -0.12) and spare capacity (in % r = -0.05, in pmol/min r = -0.11). In conclusion, this study reveals that MM changes significantly with seasons and is negatively correlated with CSA parameters and MP. Our findings indicate that the mitochondrial mass is a key parameter for determination of mitochondrial fitness. Therefore, we recommend the determination of MM during the measurement of CSA parameters for the correct interpretation and assessment of mitochondrial function.

High asymptomatic malaria among seasonal migrant workers departing to home from malaria endemic areas in northwest Ethiopia.

BACKGROUND: In Ethiopia, thousands of seasonal migrant workers travel from non-malaria or mild malaria transmission areas to malaria-endemic areas for seasonal farm activities. Most of these migrants stay in the farm areas for land preparation, plowing, planting, weeding, and harvesting for a specific period and return to their living areas. However, there is limited evidence of how seasonal migrant workers contribute to the transmission of malaria to new or less malaria transmission areas. METHODS: A cross-sectional study was conducted at the departure phase of seasonal migrant workers in the Metema district from September 2018 to October 2019. A total of 1208 seasonal migrant workers were interviewed during their departure from farm sites to their homes. The face-to-face interviews were performed using a pretested structured questionnaire. Moreover, blood samples were collected from each study participant for microscopic malaria parasite examination. The data were fitted with the logistic regression model to estimate the predictors of malaria transmission. RESULTS: At departure to home, the prevalence of malaria among seasonal migrant workers was 17.5% (15.6-19.45%). Approximately 71.80% (177/212) of the cases were Plasmodium falciparum, and 28.20% (35/212) were Plasmodium vivax. Most seasonal migrant workers 934 (77.4%) were from rural residences and highlanders 660 (55%). Most 661 (55.4%) of the migrants visited two and more farm sites during their stay at development corridors for harvesting activities. Approximately 116 (54.7%) asymptomatic malaria cases returned to the Dembia 46 (21.7%), Chilaga 46 (19.8%) and Metema 28 (13.2%) districts. CONCLUSION: In this study, asymptomatic malaria remains high among seasonal migrant workers departing to home from malaria endemic areas. This may fuel a resurgence of malaria transmission in the high lands and cause challenges to the country's malaria prevention and elimination efforts. Hence, tailored interventions for seasonal migrant workers could be in place to enhance malaria control and elimination in Ethiopia, such as asymptomatic malaria test and treat positive cases at departure and transit, and integration between malaria officers at their origin and departure for further follow-up to decrease any risk of spread at the origin.

Cellular Stress Assay in Peripheral Blood Mononuclear Cells: Factors Influencing Its Results.

Cellular stress is central to the understanding of pathological mechanisms and the development of new therapeutic strategies and serves as a biomarker for disease progression in neurodegeneration, diabetes, cancer, cardiovascular and other chronic diseases. The common cellular stress assay (CSA) based on Seahorse technology in peripheral blood mononuclear cells (PBMCs) shows inconsistent results, which prevents its use as a biomarker for the progression of chronic diseases. Therefore, the aim of this study was to investigate potential factors that affect the CSA in PBMCs. We measured the CSA parameters in PBMCs from study participants and compared the results according to the potential factors, namely, the PBMC isolation method, age, seasonal variation and the gender of the study participants. PBMCs were isolated by OptiPrep® and RobosepTM-S methods. PBMCs isolated with the OptiPrep method showed much higher extracellular acidification and higher respiration compared to Robosep-isolated cells. Moreover, OptiPrep-isolated cells showed a higher number of outliers for the proton production rate (PPR) and a high respiratory quotient, indicating impurities with other cells, such as platelets, and technical inconsistencies. PBMCs from older individuals showed higher maximal respiration, spare capacity and extracellular acidification than younger participants. Additionally, in winter, maximal respiration and spare capacity decreased. From spring until early autumn, spare capacity and maximal respiration continuously increased. Elderly males also showed higher basal respiration, spare capacity and extracellular acidification than females. In conclusion, the findings of this study clearly demonstrate that the results of CSA parameters measured in PBMCs are influenced by the PBMC isolation method, age, seasonal variation and gender. Therefore, we recommend that researchers and physicians properly interpret the results of CSA parameters in PBMCs by considering these factors. It is important to use separate CSA evaluation standards based on the isolation method, age, gender and season-dependent factors. To assess the cellular stress situation in PBMCs, both extracellular acidification and mitochondrial respiration should be taken into account. Further study of additional factors, such as mitochondrial mass, should be conducted to improve the measurement of CSA parameters for the assessment of the real mitochondrial fitness.

Prevalence of sexually transmitted infections among cervical cancer suspected women at University of Gondar Comprehensive Specialized Hospital, North-west Ethiopia.

BACKGROUND: Sexually transmitted infections (STIs) such as Human Immunodeficiency Virus (HIV), Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), and Syphilis have continued a significant public health problem, especially in areas with substandard infection prevention and control programs. It is known that STIs are largely associated with the increased occurrence of cervical cancer. To date, little is known about the burden of STIs among cervical cancer suspected women in Ethiopia. OBJECTIVES: To assess the seroprevalence of STIs and associated risk factors among cervical cancer suspected women with special emphasis on HIV, HBV, HCV, and Syphilis. METHODS: A hospital-based cross-sectional study was conducted among cervical cancer suspected women at the University of Gondar Comprehensive Specialized Hospital from February to April 2017. A pre-tested and structured questionnaire was used to collect data on patients' characteristics. The patient's serum or plasma samples were tested for HIV, HBV, HCV, and syphilis using enzyme-linked immunosorbent assay. In all aspects, the standard operational procedure was strictly followed. Data were analyzed using SPSS version 20 software and presented using tables. Statistical associations were measured using bivariate and multivariable logistic regression. A p-value of below 0.05 was considered statistically significant. RESULT: A total of 403 cervical cancer suspected women with the mean age of 42.54 (SD + 11. 24) years were enrolled in the study. The overall prevalence of STIs was 16.6% (67/403) and the prevalence of HIV, HBV, HCV, and syphilis was 36/403 (8.9%), 10/403 (2.5%), 4/403 (1%), and 29/403 (7.2%) respectively. History of multiple sexual partners (Adjusted OR = 3.02, 95%CI 1.57-5.79, P = 0.001), alcohol addiction (Adjusted OR = 2.2, 95%CI 1.07-4.5, P = 0.031), history of STI (Adjusted OR = 3.38; 95% CI: 1.76-6.47, P = 0.00) and not use of condom (Adjusted OR = 4.99; 95% CI: 1.5-16.16, P = 0.007) were significantly associated with STIs. CONCLUSION: The prevalence of STIs was generally higher among cervical cancer suspected patients. Health education is encouraged to promote awareness about the prevention of STIs.

Sero-epidemiology and associated factors of HIV, HBV, HCV and syphilis among blood donors in Ethiopia: a systematic review and meta-analysis.

BACKGROUND: Transfusion transmissible infections (TTIs) remain a major public health problem in developing countries including Ethiopia. In Ethiopia, comprehensive information about sero-epidemiology of major TTIs is lacking at the national level. Therefore, this systematic review and meta-analysis was aimed at providing the pooled estimate of human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and syphilis among blood donors in Ethiopia. METHODS: Relevant studies published until May 31, 2019 were searched through PubMed/Medline, EMBASE, SCOPUS, HINARI, Cochrane database library, Web of Science, Google Scholar and Google. The methodological quality of articles was assessed using Joanna Brigg's Institute critical appraisal checklist for prevalence and analytical studies. The pooled sero-epidemiology of HIV, HBV, HCV and syphilis were determined using the random-effects model. Heterogeneity between the studies was assessed using the I2 statistics. Publication bias was assessed by visual inspection of the funnel plot and Egger's statistics. RESULTS: A total of 7921 articles were retrieved, and 7798 were screened for eligibility after duplicates removed. Forty-nine full-text articles were assessed for eligibility; of which 45 were eligible for qualitative and quantitative synthesis: categorized as 36, 34, 31 and 23 studies for estimations of HBV, HIV, HCV and syphilis, respectively. In the random-effects model, the pooled sero-epidemiology of HBV, HIV, HCV and syphilis was 5.20, 2.83, 0.93 and 1.50%, respectively. Moreover, being a male blood donor was significantly associated with HBV and syphilis infection, whereas being a replacement blood donor was significantly associated with a high burden of HIV, HBV and HCV infections. CONCLUSION: The pooled sero-epidemiology of major TTIs among blood donors was high. Therefore, there is a need to design prevention and control strategies in a comprehensive approach to reduce the burden.

Antimicrobial resistance profile and multidrug resistance patterns of Streptococcus pneumoniae isolates from patients suspected of pneumococcal infections in Ethiopia.

BACKGROUND: Antimicrobial-resistant strains of Streptococcus pneumoniae have become one of the greatest challenges to global public health today and inappropriate use of antibiotics and high level of antibiotic use is probably the main factor driving the emergence of resistance worldwide. The aim of this study is, therefore, to assess the antimicrobial resistance profiles and multidrug resistance patterns of S. pneumoniae isolates from patients suspected of pneumococcal infections in Ethiopia. METHODS: A hospital-based prospective study was conducted from January 2018 to December 2019 at Addis Ababa city and Amhara National Region State Referral Hospitals. Antimicrobial resistance tests were performed from isolates of S. pneumoniae that were collected from pediatric and adult patients. Samples (cerebrospinal fluid, blood, sputum, eye discharge, ear discharge, and pleural and peritoneal fluids) from all collection sites were initially cultured on 5% sheep blood agar plates and incubated overnight at 37 °C in a 5% CO2 atmosphere. Streptococcus pneumoniae was identified and confirmed by typical colony morphology, alpha-hemolysis, Gram staining, optochin susceptibility, and bile solubility test. Drug resistance testing was performed using the E-test method according to recommendations of the Clinical and Laboratory Standards Institute. RESULTS: Of the 57 isolates, 17.5% were fully resistant to penicillin. The corresponding value for both cefotaxime and ceftriaxone was 1.8%. Resistance rates to erythromycin, clindamycin, tetracycline, chloramphenicol and trimethoprim-sulfamethoxazole were 59.6%, 17.5%, 38.6%, 17.5 and 24.6%, respectively. Multidrug resistance (MDR) was seen in 33.3% isolates. The most common pattern was co-resistance to penicillin, erythromycin, clindamycin, and tetracycline. CONCLUSIONS: Most S. pneumoniae isolates were susceptible to ceftriaxone and cefotaxime. Penicillin has been used as a drug of choice for treating S. pneumoniae infection. However, antimicrobial resistance including multidrug resistance was observed to several commonly used antibiotics including penicillin. Hence, it is important to periodically monitor the antimicrobial resistance patterns to select empirical treatments for better management of pneumococcal infection.

Molecular characterization of Streptococcus agalactiae isolated from pregnant women and newborns at the University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia.

BACKGROUND: Streptococcus agalctiae (Group B Streptococcus, GBS) is a perinatal pathogen and a leading cause of neonatal infections worldwide. Serotype, sequence type, clonality, antibiotic resistance genes and surface protein profiles of GBS are scarce in Ethiopia, a reason that this study was planned to investigate. . METHODS: Sixteen colonizing GBS isolates obtained from recto-vaginal swabs of pregnant women and body surfaces of newborns were further analyzed. Minimum inhibitory concentration (MIC) test, and whole genome sequence (WGS) methods were done for antibiotic susceptibility test, and molecular characterization of the isolates. RESULTS: All the GBS isolates analyzed were belonged to four capsular serotypes: II, 11/16(68.8%), V, 3/16(18.8%), Ia and VI each with 1/16(6.3%) and five sequence type (ST-2, ST-10, ST-14, ST-569 and ST-933). Sequence type-10 was the most predominant ST followed by ST-569. The five STs were grouped into the four clonal complexes (CC - 1, CC-10, CC-19, and CC-23). Different surface proteins and pili families such as ALP1, ALPHA, ALP23, PI-1 / PI-2A1, PI-1 / PI-2B, and Srr1 were detected from WGS data. All isolates were found to be susceptible to the tested antibiotics except for tetracycline in MIC and WGS test methods used. Tetracycline resistant determinant genes such as TETM and TETL / TETM combination were identified. CONCLUSION: Further studies on serotype and molecular epidemiology will provide a comprehensive data of the GBS capsular serotype and clones available in Ethiopia.

Tuberculosis and its association with CD4+ T cell count among adult HIV positive patients in Ethiopian settings: a systematic review and meta-analysis.

BACKGROUND: Tuberculosis (TB) and HV have been intertwined and makeup a deadly human syndemic worldwide, especially in developing countries like Ethiopia. Previous studies have reported different TB incidences and its association with CD4+ T cell counts among HIV positive patients in Ethiopia. Thus, the goal of this meta-analysis was, first, to determine pooled incident TB among adult HIV positive patients, and second, to assess the association between incident TB and baseline CD4+ T cell count strata's. METHODS: We searched PubMed, Cochrane library, Science Direct and Google scholar databases from June 1 to 30, 2018. The I2 statistics and Egger's regression test was used to determine heterogeneity and publication bias among included studies respectively. A random effects model was used to estimate pooled incident TB and odds ratio with the respective 95% confidence intervals using Stata version 11.0 statistical software. RESULTS: A total of 403 research articles were identified, and 10 studies were included in the meta-analysis. The pooled incident TB among adult HIV infected patients in Ethiopia was 16.58% (95% CI; 13.25-19.91%). Specifically, TB incidence in Pre-ART and ART was 17.16% (95% CI; 7.95-26.37%) and 16.24% (95% CI; 12.63-19.84%) respectively. Moreover, incident TB among ART receiving patients with baseline CD4+ T cell count < and > 200 cells/mm3 was 28.86% (95% CI; 18.73-38.98%) and 13.7% (95% CI; 1.41-25.98%) correspondingly. The odds of getting incident TB was 2.88 (95% CI; 1.55-5.35%) for patients with baseline CD4+ T cell count < 200 cells/mm3 compared to patients with baseline CD4+ T cell count > 200 cells/mm3. CONCLUSION: High incident TB among adult HIV positive patients was estimated, especially in patients with CD4+ T cell count < 200 cells/mm3. Therefore, Early HIV screening and ART initiation, as well as strict compliance with ART and increasing the coverage of TB preventive therapy to more risky groups are important to prevent the problem. TRIAL REGISTRATION: Study protocol registration: CRD42018090802.

The protective effect of isoniazid preventive therapy on tuberculosis incidence among HIV positive patients receiving ART in Ethiopian settings: a meta-analysis.

BACKGROUND: Tuberculosis (TB) and HIV makeup a deadly synergy of infectious disease, and the combined effect is apparent in resource limited countries like Ethiopia. Previous studies have demonstrated inconsistent results about the protective effect of isoniazid preventive therapy (IPT) on active TB incidence among HIV positive patients receiving ART. Therefore, the aim of this meta-analysis was, first, to determine the protective effect of IPT on active tuberculosis incidence, and second, to assess the pooled incidence of active TB among HIV positive patients taking ART with and without IPT intervention in Ethiopia. METHODS: PubMed, Google scholar and Cochran library databases were searched from April 1 to 30, 2018. Two independent authors explored and assessed studies for eligibility, and extracted data based on predefined criteria. Studies that reported TB incidence among HIV positive patients taking ART in Ethiopia with and without IPT concomitant intervention, and with a clear stratified data on the incidence of TB based on the duration of IPT intervention were selected. A random effects model was used to estimate risk ratios and the pooled incident TB with the respective 95% confidence intervals. RESULTS: We identified 7 suitable studies in this analysis. Accordingly, IPT reduced the risk of TB incidence by 74%, risk ratio (RR) 0.26 (95% CI; 0.16-0.43%), compared to no IPT group. Moreover, IPT for 12 months reduced incident TB by 91% (RR: 0.09, 95% CI: 0.04 to 0.21), whereas 6 months IPT averted TB incidence by 63% (RR: 0.37, 95% CI: 0.26 to 0.52). The overall pooled incident TB among HIV infected patients receiving ART was 10.30% (95% CI; 7.57-13.02%). Specifically, incident TB among study cohorts with and without IPT was 3.79% (95% CI; 2.03-5.55%) and 16.32% (95% CI; 11.57-21.06%) respectively. CONCLUSION: IPT reduced the risk of incident TB among HIV positive patients receiving ART in Ethiopian settings. Moreover, the duration of IPT intervention has effect on its protective role. Thus, scaling up the isoniazid preventive therapy program and its strict compliance is necessary to avert HIV fueled tuberculosis. STUDY PROTOCOL REGISTRATION: CRD42018090804.

Streptococcus agalactiae maternal colonization, antibiotic resistance and serotype profiles in Africa: a meta-analysis.

BACKGROUND: Maternal rectovaginal colonization with Streptococcus agalactiae (Group B Streptococcus or GBS) is the most common route for the GBS disease in the perinatal period. The knowledge of maternal colonization, antibiotic resistance and serotype profiles is substantially needed to formulate the broad vaccine. However, it has not been estimated in Africa. This meta-analysis was aimed to determine the pooled prevalence of colonization, antibiotic resistance and serotype profiles of GBS reported in Africa. METHODS: Potentially relevant studies from 1989 to 31th January, 2019 were retrieved from the Medline/PubMed, EMBASE, HINARI online databases, periodicals and by requesting authors. Unpublished studies retrieved from grey literature through Google and Google Scholar. Pooled estimates were calculated using the random effect model. Subgroup analysis was done to investigate the burden of colonization across sub-regions, sampling site and countries. Summary estimates were presented using words, Forest plots and Tables. Heterogeneity was assessed using the I2 statistic. RESULTS: Eighty-three articles were assessed, of which 57 studies conducted in five sub-regions with 21 countries (22,206 pregnant women) met pre-specified inclusion criteria. The overall estimate of recto-vaginal colonization was 19.3% (95% CI 16.9, 21.7). The highest estimate was observed in Southern Africa, 23.8% (95% CI 18.7, 28.9), followed by Northern Africa, 22.7% (95% CI 18.2, 27.2) while the lowest was driven from the Eastern Africa, 15.4% (95% CI 12.1, 18.7). Considerable heterogeneity across and within regions, sampling site, screening methods and countries (I2 > 75%); and the publication bias were observed (p = 0.031). GBS showed the highest resistance to tetracycline. Resistance to penicillin, amoxicillin, chloramphenicol, ampicillin, ceftriaxone, ciprofloxacin, erythromycin, vancomycin and clindamycin also observed. The V, III, Ia, Ib, and II serotypes altogether were accounted 91.8% in the African studies. CONCLUSIONS: The pooled estimate of the maternal colonization with GBS was 19.3% which is equivalent with other many primary and review reports worldwide. The most antibiotic resistance estimate was recorded in the tetracycline followed by penicillin. Five serotypes were the most prevalent in Africa and more data on the antibiotic résistance and serotype distribution patterns are needed from developing countries to devise the effective preventive measures. In addition, the antibiotic susceptibility test methods used in the Africa shall be assessed for its quality. Trial registration Prospero Registration Number CRD42018094525.

Streptococcus agalactiae from Ethiopian pregnant women; prevalence, associated factors and antimicrobial resistance: alarming for prophylaxis.

BACKGROUND: Maternal Streptococcus agalactiae (Group B Streptococcus, GBS) colonization rates and its antibiotic resistance patterns provide important information useful in guiding prevention strategies. There is a paucity of evidence about GBS in the Amhara National Regional State, Ethiopia. OBJECTIVE: To determine colonization prevalence, associated risk factors, and antibiotics resistance including inducible clindamycin resistance patterns of GBS among Ethiopian pregnant women. METHODS: A prospective cross-sectional study was conducted from 1st December 2016 to 30th November 2017 at the University of Gondar Referral hospital delivery ward. Combined recto-vaginal swabs were collected from 385 pregnant women and analyzed at the University of Gondar Bacteriology Laboratory by using LIM broth and 5% defibrinated sheep blood agar culture methods. Isolates were identified by using colony morphology, gram reaction, hemolysis, and CAMP test. Antibiotic susceptibility test was done using the disc diffusion method. Double disc diffusion method was used to identify inducible clindamycin resistance isolates. Data were analyzed by SPSS version 20 software. p ≤ 0.05 was considered as statistically significant. RESULTS: The overall prevalence of maternal GBS colonization was 25.5% (95% CI 21-29.5%). Experiencing meconium stained amniotic fluid (AOR = 3.018, 95% CI 1.225, 7.437), and longer duration of premature rupture of membrane (AOR = 1.897, 95% CI 1.014, 3.417) were statistically significant to maternal colonization. Furthermore, GBS resistant to 0 (8.2%), 1 (25.5%) and 3 (39.8%) or more antibiotics were identified. A D-test showed 15.2% inducible clindamycin resistant GBS. Constitutive macrolide lincosamide-streptograminB, L-, and M-phenotypes were also detected. CONCLUSIONS: Maternal GBS colonization rate in this study was higher compared to the previous reports in Ethiopia. This much prevalence and antibiotics resistance results are the clue to which attention shall be given to this bacterium during management of pregnant women and the newborns.

Prevalence and associated factors of treatment failure among HIV/AIDS patients on HAART attending University of Gondar Referral Hospital Northwest Ethiopia.

BACKGROUND: The initiation of highly active antiretroviral therapy (HAART) plays a significant role in the clinical management of HIV infected people by preventing morbidity and mortality. This benefit becomes, the most terrible when treatment failure develops. Thus, this research aims to assess the prevalence and associated factors of treatment failure among HIV/AIDS patients on HAART attending University of Gondar Referral Hospital Northwest Ethiopia. RESULTS: Patients on ART with a minimum of 6 months and up to 12 years of treatment were being enrolled. The prevalence of treatment failure, immunological failure and virological failure among people living with HIV/AIDS attending University of Gondar referral hospital were 20.3, 13.2, and 14.7%, respectively. Patients who had no formal education (Adjusted odds ratio (AOR): 3.8; 95% CI, 1.05-13.77), primary level education (AOR: 4.2; 95% CI, 1.16-15.01) and duration on ART < 6 years (AOR: 2.1; 95%CI, 1.12-3.81) were a significant risk factor. However, initial adult regimen D4T +  3TC+ EFV (AOR: 0.025; 95% CI, 0.002-0.36), AZT +3TC + NVP (AOR: 0.07; 95% CI, 0.01-0.71), AZT +  3TC + EFV (AOR: 0.046; 95% CI, 0.004-0.57) andTDF+3TC + EFV (AOR: 0.04; 95% CI, 0.004-0.46) were significantly protective for treatment failure. CONCLUSIONS: Timely and early identification of associated factors and monitoring antiretroviral therapy treatment failure should be done to enhance the benefit and to prevent further complication of the patients. It is preferable to initiate ART using any one of the following ART regimens: AZT +3TC + NVP, AZT + 3TC + EFV and TDF + 3TC + EFV to prevent treatment failure. Since the prevalence of this treatment failure and its associated factor may be different from other ART centers and community in Ethiopia, further national representative institutional based cross-sectional researches are needed across all ART centers of Ethiopia in order to determine the prevalence of treatment failure and its associated factors.

A study of clinical, hematological, and biochemical profiles of patients with dengue viral infections in Northwest Ethiopia: implications for patient management.

BACKGROUND: Dengue is one of the most common arboviral diseases with increased outbreaks annually in tropical and subtropical areas. In Ethiopia, there are no data regarding clinical, hematological and biochemical parameters which are very important in the clinical management of dengue patients. Hence this study was carried out to provide the first baseline data of clinical, hematological and biochemical profiles of patients infected with dengue virus. METHODS: A cross-sectional study was carried out among febrile patients in northwest Ethiopia from March 2016 to May 2017. Blood samples were collected from dengue presumed cases and tested against dengue specific IgM antibody by enzyme-linked immunosorbent assay (ELISA). Those study participants who fulfilled the inclusion criteria were enrolled in the study. Clinical examination findings were recorded, hematological and biochemical parameters tests were done. RESULTS: During the study period, a total of 102 dengue cases were included in the study. Of these, there were 16 (15.7%) children and 86 (84.3%) adults between 1 and 76 year age. The most common clinical presentations followed by fever (100%) were a headache 89 (87.3%), myalgia 82 (80.4%), nausea/vomiting 71 (69.6%). The common hematological findings were thrombocytopenia 61 (59.8%), followed by anemia 45 (44.1%) and leucopenia 27 (26.5%) and the elevated levels of biochemical parameters were AST 46 (45.1%) and ALT in 18 (17.6%). CONCLUSIONS: This study highlights the most common clinical and laboratory profiles of dengue viral infections that could alert physicians to the likelihood of dengue virus infections in the study area.

Newborn colonization and antibiotic susceptibility patterns of Streptococcus agalactiae at the University of Gondar Referral Hospital, Northwest Ethiopia.

BACKGROUND: Group B Streptococcus (GBS) that asymptomatically colonizing the recto-vaginal area of women is the most important cause of neonatal colonization. There is paucity of evidence about newborn colonization with GBS in Ethiopia. Thus, this study was aimed to determine the prevalence of newborn colonization with GBS, antibiotic susceptibility patterns of the isolates and associated risk factors at the University of Gondar Referral Hospital in Northwest Ethiopia METHODS: A prospective cross sectional study was conducted from December 2016 to November 2017. A total of 1,155 swabs from nasal, ear and umbilical areas of the newborns were collected from the 385 newborns. Identifications of the isolates and antibiotic susceptibility testing were done by using conventional methods. RESULTS: Sixty two (16.1%, 95% CI: 12.2% - 20%) of the newborns were colonized by GBS. Seven percent of the total specimens were positive for GBS. The antibiotics susceptibility rates of GBS (average of the three body sites tested) were 95.1%, 89.6%, 88.9%, 85.7%, 85.3%, 81.3%, 76.9%, 76.1%, 73.8%, and 34.4% to ampicillin, penicillin, ciprofloxacin, chloramphenicol, vancomycin, azitromycin, erythromycin, clindamycin, ceftriaxone, and tetracycline, respectively. A multilogistic regression analyses were shown that the newborns that were from mothers whose education status was below tertiary level, and newborns from mothers who were: being employed, being nullipara and multigravida were at risk for colonization with GBS. CONCLUSION: Prevalence of neonatal colonization with GBS was higher than it was reported in three decades ago in Ethiopia. Ciprofloxacin, chloramphenicol, vancomycin and azithromycin were identified as the drug of choice next to ampicillin and penicillin.

A standardised method for interpreting the association between mutations and phenotypic drug resistance in Mycobacterium tuberculosis.

A clear understanding of the genetic basis of antibiotic resistance in Mycobacterium tuberculosis is required to accelerate the development of rapid drug susceptibility testing methods based on genetic sequence.Raw genotype-phenotype correlation data were extracted as part of a comprehensive systematic review to develop a standardised analytical approach for interpreting resistance associated mutations for rifampicin, isoniazid, ofloxacin/levofloxacin, moxifloxacin, amikacin, kanamycin, capreomycin, streptomycin, ethionamide/prothionamide and pyrazinamide. Mutation frequencies in resistant and susceptible isolates were calculated, together with novel statistical measures to classify mutations as high, moderate, minimal or indeterminate confidence for predicting resistance.We identified 286 confidence-graded mutations associated with resistance. Compared to phenotypic methods, sensitivity (95% CI) for rifampicin was 90.3% (89.6-90.9%), while for isoniazid it was 78.2% (77.4-79.0%) and their specificities were 96.3% (95.7-96.8%) and 94.4% (93.1-95.5%), respectively. For second-line drugs, sensitivity varied from 67.4% (64.1-70.6%) for capreomycin to 88.2% (85.1-90.9%) for moxifloxacin, with specificity ranging from 90.0% (87.1-92.5%) for moxifloxacin to 99.5% (99.0-99.8%) for amikacin.This study provides a standardised and comprehensive approach for the interpretation of mutations as predictors of M. tuberculosis drug-resistant phenotypes. These data have implications for the clinical interpretation of molecular diagnostics and next-generation sequencing as well as efficient individualised therapy for patients with drug-resistant tuberculosis.

FIND Tuberculosis Strain Bank: a Resource for Researchers and Developers Working on Tests To Detect Mycobacterium tuberculosis and Related Drug Resistance.

The spread of multidrug-resistant (MDR) tuberculosis (TB) and extensively drug-resistant (XDR) TB hampers global efforts in the fight against tuberculosis. To enhance the development and evaluation of diagnostic tests quickly and efficiently, well-characterized strains and samples from drug-resistant tuberculosis patients are necessary. In this project, the Foundation for Innovative New Diagnostics (FIND) has focused on the collection, characterization, and storage of such well-characterized reference materials and making them available to researchers and developers. The collection is being conducted at multiple centers in Southeast Asia, South America, Eastern Europe, and soon the sub-Saharan Africa regions. Strains are characterized for their phenotypic resistances and MICs to first-line drugs (FLDs) and second-line drugs (SLDs) using the automated MGIT 960 system following validated procedures and WHO criteria. Analysis of resistance-associated mutations is done by whole-genome sequencing (WGS) using the Illumina NextSeq system. Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat analysis and WGS are used to determine strain lineages. All strains are maintained frozen at -80°C ± 10°C as distinct mother and daughter lots. All strains are extensively quality assured. The data presented here represent an analysis of the initial part of the collection. Currently, the bank contains 118 unique strains with extracted genomic DNA and matched sputum, serum, and plasma samples and will be expanded to a minimum of 1,000 unique strains over the next 3 years. Analysis of the current strains by phenotypic resistance testing shows 102 (86.4%), 10 (8.5%), and 6 (5.1%) MDR, XDR, and mono/poly resistant strains, respectively. Two of the strains are resistant to all 11 drugs that were phenotypically tested. WGS mutation analysis revealed FLD resistance-associated mutations in the rpoB, katG, inhA, embB, embA, and pncA genes; SLD resistance in the gyrA, gyrB, rrs, eis, and tlyA genes; and ethionamide resistance in the ethA genes. Most important lineages are represented in the bank, and further collections have been initiated to increase geographic and lineage diversity. The bank provides highly characterized and high-quality strains as a resource for researchers and developers in support of the development and evaluation of new diagnostics and drug resistance detection tools.

Smear positive pulmonary tuberculosis and associated risk factors among tuberculosis suspects attending spiritual holy water sites in Northwest Ethiopia.

BACKGROUND: Tuberculosis (TB) remains one of the world's deadliest communicable diseases. In Ethiopia, tuberculosis patients have different pattern of health care seeking behavior. They usually adopt other approaches like traditional healers and spiritual holy water sites before consulting public health facilities. This study was aimed to assess the prevalence of smear positive pulmonary tuberculosis and associated risk factors among tuberculosis suspects attending spiritual holy water sites. METHODS: A cross-sectional study was conducted from February 01, 2015 to March 30, 2015 in seven selected holy water sites in Northwest Ethiopia. During the study period, a total of 1384 adult holy water users were screened for PTB symptoms. A total of 382 pulmonary tuberculosis suspects participated in the study. Socio-demographic data were collected using a semi-structured questionnaire. Spot-morning-spot sputum specimens were collected and examined for acid fast bacilli using Auramine O fluorescence staining technique. Smear positive sputum samples were tested by GeneXpert MTB/RIF assay for rifampicin resistance. Descriptive statistics, binary and multivariate logistic regression analysis were employed using SPSS-16 software. RESULTS: The prevalence of smear positive pulmonary tuberculosis was 2.9% with point prevalence of 795/100, 000 holy water users. History of contact with tuberculosis patient (AOR = 9.174, 95% C.I = 2.195-38.34) and the number of family members > 5 per household (AOR = 9.258, 95% C.I = 1.14-74.97) were significantly associated with smear positive pulmonary tuberculosis. Rifampicin resistance was not detected from all smear positives by GeneXpert MTB/RIF assay. CONCLUSIONS: The prevalence of smear positive pulmonary tuberculosis in spiritual holy water sites was 7.4 fold higher than the general population. History of contact with active tuberculosis patients and increased family size were significantly associated with smear positive pulmonary TB. The national tuberculosis program should consider spiritual holy water sites as potential foci for TB transmission and plan regular survey and health education in holy water sites for effective TB prevention and control in the country.

Multidrug resistant tuberculosis in Ethiopian settings and its association with previous history of anti-tuberculosis treatment: a systematic review and meta-analysis.

BACKGROUND: Efforts to control the global burden of tuberculosis (TB) have been jeopardized by the rapid evolution of multi-drug resistant Mycobacterium tuberculosis (MTB), which is resistant to at least isoniazid and rifampicin. Previous studies have documented variable prevalences of multidrug-resistant tuberculosis (MDR-TB) and its risk factors in Ethiopia. Therefore, this meta-analysis is aimed, firstly, to determine the pooled prevalence of MDR-TB among newly diagnosed and previously treated TB cases, and secondly, to measure the association between MDR-TB and a history of previous anti-TB drugs treatment. METHODS: PubMed, Embase and Google Scholar databases were searched. Studies that reported a prevalence of MDR-TB among new and previously treated TB patients were selected. Studies or surveys conducted at national or sub-national level, with reported MDR-TB prevalence or sufficient data to calculate prevalence were considered for the analysis. Two authors searched and reviewed the studies for eligibility and extracted the data in pre-defined forms. Forest plots of all prevalence estimates were performed and summary estimates were also calculated using random effects models. Associations between previous TB treatment and MDR-MTB infection were examined through subgroup analyses stratified by new and previously treated patients. RESULTS: We identified 16 suitable studies and found an overall prevalence of MDR-TB among newly diagnosed and previously treated TB patients to be 2% (95% CI 1% - 2%) and 15% (95% CI 12% - 17%), respectively. The observed difference was statistically significant (P < 0.001) and there was an odds ratio of 8.1 (95% CI 7.5-8.7) for previously treated TB patients to develop a MDR-MTB infection compared to newly diagnosed cases. For the past 10 years (2006 to 2014) the overall MDR-TB prevalence showed a stable time trend. CONCLUSIONS: The burden of MDR-TB remains high in Ethiopian settings, especially in previously treated TB cases. Previous TB treatment was the most powerful predictor for MDR-MTB infection. Strict compliance with anti-TB regimens and improving case detection rate are the necessary steps to tackle the problem in Ethiopia.