Browsing by Author "Asrat Daniel"

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    Analysis of Serum for Antibodies to Cytomegalovirus in Individuals with Schizophrenia and Bipolar Disorder
    (Addis Ababa University, 2010-02) Yared Tedla ; Woldeamanuel Yimtubezinash; Asrat Daniel; Shibre Teshome; Assefa Abrham
    Background: Several evidences suggest that Cytomegalovirus (CMV) may play anetiological role in some case of schizophrenia and bipolar disorder. Studies have reported that some individuals experiencing initial episodes of major mental disorders have increased levels of immunoglobulin G(IgG) to CMV. Objective: To determine the magnitude of CMV infection in individuals with schizophrenia, bipolar disorder and healthy controls by using serologic diagnostic methods. Materials and Methods: A case control study conducted from March to May of 2009. A total of 844 serum samples were analysed for the presence and level of IgG to CMV using enzyme linked immunosorbent assay (ELISA).Serawere collected from 216 individuals with schizophrenia, 199with bipolar disorder and 429close relatives and their healthy controls, recruited from Butajira district.Seropositivity summarized using percentage and antibody level was summarized using mean. Logistic regression was used for cross tabulated data and linear regression for comparing antibody level among groups. All test of significance was two tailed and ,p < 0.05 is considered significant. Results:99.3% of tested serawere positive for IgG to CMV. However, serum levels of IgGto CMV in individuals with schizophrenia[β = 0.21; 95% CI (0.03, 0.38)]and bipolar disorder [β = 0.20; 95% CI (0.02, 0.37)]were higher compared to healthy controls. Younger individuals with schizophrenia (<20 years) had higher level of IgG to CMV compared to matched healthy controls[β = 0.64;95% CI (0.10, 1.19)]. Conclusion: This study provided serologic evidence that infection with CMV may be associated with some cases of schizophrenia and bipolar disorder. Additional studies should be directed at further analysis of antibodies to CMV in the sera and CSF of individuals with recent onset of psychosis.
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    Assessment of Bacterial Profile and Antimicrobial Susceptibility Pattern of Catheter-Associated Urinary Tract Infections in Comparison with non-Catheterized Urinary Tract Infections in Jimma University Hospital, Southwest Ethiopia
    (Addis Ababa University, 2005-06) Teshager Lule; Asrat Daniel
    Urinary tract infections (UTIs) are among the most common bacterial infection.They have become the most common hospital acquired infections, accountingfor as many as 35% of nosocomial infections, and are usually associated withcatheterization. The aim of this study is to assess the bacterial profile andantimicrbial resistance pattern of catheter associated urinary tract infection incomparison with non-catheter associated UTI. One hundred and twenty urinespecimens (30 from catheterized patients and 90 from non-catheterized patients with symptoms of UTI) were screened for the presence of significant bacteruria from January to March 2005. Bacteriological screening of catheterized urine and clean catch mid-stream urine revealed that 13/30 (43.3%) and 20/90 (22.3%) had significant bacteriuria, respectively (P<0.05). All samples taken from patients who have been catheterized for 2 weeks or more showed significant bacteruria. In both group the rate of infection was higher in female patients. Older patients (>50years old) were found to be the most affected age group (46%) among catheterized patients whereas most of the non-catheterized patients with significant bacteruria belonged to age group 11-12 years. Among catheterized patients, Escherichia coli and Klebsiella species were found to be the most frequently isolated pathogens (each of them accounts 23 %) followed by coagulase negative Staphylococci (15 %). The most frequently isolated species from non-catheterized patients was Klebsiella (40 %) followed by E. coli (30 %). Resistance rates (56.3 % to 100%) to Ampicillin, amoxicillin, cephalexin, carbenicillin, and Trimethoprime /sulfamethoxazole were observed in all organisms isolated from both groups. In addition, resistance rates to gentamicin, nitrofurantoin, and nalidixic acid were observed in all bacteria isolated from catheterized patients. In general, in this study, high level multidrug resistance in both groups indicated that it is time to reconsider the empirical use of the commonly used antimicrobial agents in Ethiopia. Particularly, increasing vii resistance in pathogens isolated from catheterized patients is frustrating. Because the most important risk factors for UTI is duration of catheterization indwelling urinary catheterization should be avoided or at least minimized
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    Assessment of Specific Immune Responses to Tuberculosis in HIV Infected Patients Before and During Highly Active Antiretroviral Treatment (HAART)
    (Addis Ababa University, 2009-03) Berhanu Tesfaye; Asrat Daniel; Woldeamanuel Yimtubezinash
    BACKGROUND: HAART greatly reduces the risk of developing tuberculosis in HIVinfected persons. However, individuals who initiate HAART may still be reported as having TB, either because they are developing active TB due to persistent immunodeficiency or because they have sub clinical TB that becomes apparent in the immune reconstitution inflammatory syndrome (IRIS). Despite the severity of the problem, little information is available on the extent to which HAART restores TB specific immune response or CD4 cell and characteristics of patients who develop TB while taking HAART and proportion of immune reconstitution syndrome associated with TB. OBJECTIVES: To assess specific immune response to tuberculosis in HIV-1 infected patients before and during HAART and to assess the characteristics of patients who developed ART associated TB and to determine the incidence rate of TB IRIS. METHODS: In a longitudinal prospective cohort study, 177 study participants naive to HAART were enrolled and followed for six months after starting HAART at ALERT Hospital, Addis Ababa Ethiopia. The study period was from June 2006 to September 2008. Blood samples were collected before initiation of HAART (at baseline), at 3rd and 6th month of HAART and at occurrence of suspected TB IRIS. T lymphocyte sub set enumerated and the immune response to tuberculosis was assessed in vitro, using ELISPOT assay in PBMC stimulated with TB antigens (PPD, ESAT-6, and CFP-10), and in vivo using tuberculin skin test (TST) at the specified time points. RESULTS: Recovery of TB specific immune response: The proportion of TST positive responses increased significantly from baseline 17.5% to 26.8% at 3rd month and to 28.9% at the 6th month (p= 0.02). TST response increased significantly among groups with CD4 cell count <50 cells/mm3 when the group is stratified according to baseline CD4 cell count and among group with active TB at baseline investigation when the group is stratified according to baseline TB status. Quantitative IFN-γ ELISPOT response to TB antigens (PPD, ESAT-6 and CFP-10) also increased after HAART in the whole cohort. Median SFC/million to PPD xiii and ESAT-6 significantly increased among group with baseline CD4<50 cells/mm3 when the group is stratified according to baseline CD4 cell count. Although the difference was not statistically significant (except for quantitative ELSPOT response to PPD), the proportion of TST response and median SFC/million ELISPOT response to TB antigens after HAART were also higher in those with baseline CD4 ≥100 cells/mm3 than < 100 cells/mm3. ART associated TB, unmasking and paradoxical TB IRIS: the incidence rate of ART associated TB in our cohort was 12.7 cases /100 PY during the first 6 months of HAART. The majority (62.5%) of TB cases occurred within the first month of HAART. Baseline TST response > 5 mm induration, baseline hemoglobin < 12 gm/dl and being male were the strongest predictors of occurrence of TB during HAART. Only 3/8 (37.5%) episodes of TB were presented as unmasking tuberculosis IRIS. The proportion of paradoxical IRIS was 9.6 % (3/31) and the incidence rate was 22.5 cases per 100 persons per year. CONCLUSIONS AND RECOMMENDATIONS: The majority of the study participants attained CD4 cell counts above the zone of severe immunodeficiency within the first 3 months of treatment. However, the relative risk of attaining CD4 cell > 200 cells/mm3 decreased significantly with decreasing baseline CD4 cell count. Thus, individuals who initiated HAART with CD4 < 50 cells/mm3 are likely to have high risk of morbidity and mortality for longer period. This underscores the need of designing strategy to identify and treat patients before they get into a stage of profound CD4 lymphcytopenia. Our study indicates that TB specific T cell repertoire may not be totally lost in HIV infected individuals with CD4 < 50 cells/ mm3 and their levels can still be improved by HAART. Our findings provide useful baseline information for further research to assess overall potential of HAART in the restoration of immunity to tuberculosis. Finally, in this study, we observed a high rate of tuberculosis in our cohort during the initial months of HAART indicating the need for careful screening for TB before initiation of HAART. More sensitive and specific diagnostic methods for TB may assist in early diagnosis. In addition, our study confirmed that a sub set of patients who developed TB during HAART manifested as IRIS. Key words: Tuberculosis, CD4 cell count, IRIS, HAART, immune response and Ethiopia
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    Bacterial Profile and Drug Susceptibility Pattern of Urinary Tract Infection in Pregnant Women Attending Antenatal Care at Mekelle Hospital, Mekelle, Northern Ethiopia
    (Addis Ababa University, 2014-12) Tsegay Ephrem; Asrat Daniel; Woldeamanuel Yimtubezenash
    Background: Urinary tract infection (UTI) in pregnancy is associated with significant morbidity for both the mother and the baby. However, little is known about UTI in pregnancy in the study area. Antimicrobial resistance among the pathogens that cause UTI is also increasing and is a major health problem in the treatment. Hence, proper investigation and prompt treatment are needed to prevent serious life threatening condition and morbidity due to UTI that can occur in pregnant women. Objectives: To identify the prevalent bacterial isolates that cause UTI and assess their antibiotic susceptibility pattern among symptomatic and asymptomatic pregnant women attending antenatal care in Mekele Hospital, Tigray Region. Methods: A cross sectional study was carried out from January to August 2014 on a total of 168 pregnant women with and without symptoms of UTI that attended antenatal care at Mekelle Hospital. Mid-stream urine samples were collected and inoculated onto Cystine Lactose Electrolyte Deficient medium (CLED). Colony counts yielding bacterial growth of ≥105cfu/ml is regarded as significant bacteriuria. Pure isolates of bacterial pathogen were characterized by colony morphology, gram-stain, and standard biochemical procedures. A standard method of agar disc diffusion susceptibility testing method was used to determine susceptibility patterns of the isolates. Results: In this study, the overall prevalence of UTI was 11.9%. Of this bacteriological screening of midstream urine specimens showed that 11.3% and 15.4%, had significant bacteriuria in asymptomatic and symptomatic group respectively. Prevalence of UTI was significantly associated with previous history of catheterization and urinary tract infection (p<0.05). Escherichia coli was the most frequently isolated organism 6(30%) followed by coagulase negative staphylococcus 5(25%), Staphylococcus aureus 3 (15%), and Klebsiella pneumoniae 3 (15%). Gram negative and Gram positive bacteria accounted for (60) and (40%) respectively. Gram-negative isolates showed resistance rate of 100% to ampicillin and resistance against ciprofloxacin, norfloxacin, gentamycin, amoxicillinclavulnic acid, trimethoprim/sulfamethoxazole and chloramphenicol ranged from 25- 50%. However, all Gram negative bacterial isolates revealed low level of resistance VIII (16.7%) against nitrofurantoin and ceftriaxone. The rates of susceptibility of Grampositives to ceftriaxone, doxycycline, amoxicillin-clavulnic acid and vancomycin range from 62.5% - 100%. And they showed resistance rate of 75% and 87.5% to penicillin and ampicillin, respectively. Multiple drug resistance (resistance to two or more drugs) was observed in 90 % of the isolates. Conclusions and Recommendations:- Significant bacteriuria has been observed from both symptomatic and asymptomatic pregnant women. Majority of the isolates were resistant to the commonly prescribed antibiotics. This calls for an early screening of all pregnant women for UTI and those found to be infected need to be treated with an appropriate drug to avoid complications. Keywords: Bacterial profile, antibiotic resistance, pregnancy, Mekelle, Ethiopia
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    Bacterial Urinary Tract Infection among Adult Renal Transplant Recipients at St. Paul’s Hospital Millennium Medical College, Addis Ababa, Ethiopia
    (Addis Ababa University, 2019-04) Kiros Teklehaimanot; Asrat Daniel
    Background: Although significant advances have been made in surgical techniques and immunosuppression for renal transplantation, urinary tract infections continue to be a major public health problem globally. Post transplantation urinary tract infection complications including asymptomatic bacteriuria, cystitis, and pyelonephritis are the most common form of bacterialinfection following renal transplantation. It could result in graft function impairment and death. Objectives: The study aimed to investigate the prevalence of bacterial isolates that cause urinary tract infections, assess antibiotic susceptibility pattern among symptomatic and asymptomatic renal transplant recipients attending for health care at St. Paul’s Hospital Millennium Medical College. Materials and Methods: A hospital-based cross-sectional study was carried out from December 2017 to August 2018 among 74 renal transplant recipients. The sociodemographic characteristics were collected using structured questionnaires. A first morning voided clean-catch mid-stream urine specimens were collected and 0.001ml inoculated onto a blood and MacConkey agar plates following the standard bacteriological protocols. It was incubated aerobically at 35–37°C for 18–24 hours. Cultural characteristics and series of biochemical tests were used for identification. Results: Significant bacteriuria was found in 11/74 (14.9%), 95%CI =8.2-24.7) of the patients. The prevalence among females 6/32(18.75%) was higher than males 5/42(11.9%) without significant association (COR=2.09, 95%CI=1.04-8.45, P=0.253). UTI was higher in the age group between 35–49 years old (19.3%. Age was statistically significant and stronger independent associated risk factor with crude odds ratio=3.67, 95%CI=2.89-20.07 and 0.003, respectively. The most prevalent bacteria isolates were Escherichia coli 2(18.2%), Staphylococcus aureus 2(18.2%), Acinetobacter spp. 2(18.2%), Enterococcus spp. 2(18.2%), Coagulase-negative Staphylococci 2(18.2%)] followed by Portus mirabilis 1(9.1%).The majority (80%) of Gram-negative bacteria were resistant to ciprofloxacin, chloramphenicol, and trimethoprim /sulfamethoxazole. Multi drug resistance pattern was shown in 82% of the isolates. Conclusion and Recommendations: The overall prevalence of UTI in the study was low with a prevalence of 14.9%. Better to establish routine urine cultures especially in the first 6-12 months.
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    Bacteriology of Open Fracture Wounds in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia
    (Addis Ababa University, 2009-05) Abraham Yishak; Asrat Daniel
    Open fractures are those exposed to the out side environment through a skin wound. They are at risk of complications such as infected non-union and other co-morbid conditions. Sixty to seventy percent of compound fractures are believed to be contaminated with bacteria at the time of injury from both skin and environment. Infection of open fractures depends on the microbial and host factors. In Ethiopia, a high incidence of open fracture wound infection is suspected though the magnitude of the problem is not known. No documented report on bacterial isolates from open fracture wounds and their drug resistance pattern. During a period of November 2007 and May 2008, a cross-sectional prospective study was conducted to determine the bacteriology of open fracture wounds of 191 informed and consented patients who visited the orthopedic surgery department of Tikur Anbessa University Hospital, Addis Ababa, Ethiopia. The modified Gustilo and Anderson (G-A)grading of open fractures based on severity and extent of soft-tissue injury was used to categorize the open fractures. The clinical features of the open fracture wounds were assessed and wound swab specimens were collected using Levine’s technique from each patient. All of the wound specimens were processed for microscopic examination, culture and sensitivity testing. The causes of the fractures varied, but most of the open fractures were caused by car accidents (37.2%) and occurred in lower extremities bones (60.0%). Ofthe 191 patients, 82.7% were males and 17.3% were females (p < 0.05) resulting in an overall male to female ratio of 4.8:1. The average age of the patients was 31.55 years (age range 4 to 75 years). According to G-A grading, 23.0% of the fractures were grade I; 41.5% were grade II; 14.0% were grade IIIA; 5.5% were grade IIIB and 16.0% were grade IIIC. Of the 200 wound specimens examined by gram stain, 30.5% were positive for the presence ofbacteria. Out of the 200 wound specimens cultured, 82 (41%) were positive for bacteria. Of the culture-positive wounds, 51.2% showed mono-microbial growth and 48.8% showed polymicrobial growth. In general, a total of 162 bacterial pathogens were isolated from the open fracture wounds sampled. Staphylococcus aureus was the dominant isolate (14.8%) followed by Acinetobacter spp. (11.4%). The gram-positive and gram-negative bacteriaaccounted for 34.0% and 66.0%, respectively (p< 0.05). All gram-positive bacterial isolates showed low level of resistance (<60%) to all antibiotics tested except for ampicillin and penicillin to which they showed intermediate level of resistance (60-80%). Most gramviii positive isolates, 29/55 (52.7%) showed multiple drug resistance (resistance to three or more drugs). All Clostridium spp. were susceptible to tetracycline, doxycycline, and kanamycinand showed low level of resistance (<60%) against chloramphenicol, clindamycin and penicillin. All gram negative bacterial isolates showed low level of resistance (<60%) to all antibiotics tested except for ampicillin and amoxicillin (60-80%, intermediate level resistance). Fifty-one percent of the gram negative bacterial isolates were identified as multiple drug resistant. In conclusion, the present study showed that road traffic accident was the commonest cause of open fractures. Most fractures occurred in lower extremities. Staphylococcus aureus was the commonest isolate associated with open fracture wound infection. Gentamicin, ciprofloxacin and norfloxacin were the most effective drugs against the tested gram positive and gram negative bacteria. The findings of this study will give valuable information for establishing empiric therapeutic approaches for the management of open fracture wound infections. In addition, the findings underscore the need for routine microbiological investigation of open fracture wounds and monitorring antimicrobial resistance pattern for the use of prophylactic and therapeutic antimicrobials. Key words: Open fracture wounds, Bacterial isolates, Antimicrobial susceptibility testing
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    Burden of Campylobacter Species on Livestock Owning Households in Per-urban Addis Ababa,Ethiopia:a one Health Approach.
    (Addis Ababa University, 2020-06) Chala Gemechu; Asrat Daniel
    Background: Campylobacter is the most common infectious causes of diarrhea and acute gastroenteritis globally, and has been recognized as a significant zoonotic agent. Numerous animals serve as reservoirs and a potential source Campylobacter species for food, water and environmental contamination. The rising of antimicrobial resistance among st Campylobacter isolates, is a significant global issue. Objective (s): The study was aimed to highlight the importance of livestock, poultry and water in peri-urban Addis Ababa, as sources of zoonotic Campylobacter species and antimicrobial susceptibility of isolated strains. Methods: A cross-sectional study was conducted from December 2018 till April 2020 to collect and process 519 samples including feces from livestock, poultry, human and water samples form livestock keeping households of the peri-urban Addis Ababa. Data on households’ socio demographics, knowledge and practices regarding livestock production was collected using a structured questionnaire. Results: Of the 519 tested samples, 67 (13%) were positive for Campylobacter species, in which 10.1%, 18.5%, 13%, 13.3%, 7.1% and 10.5% were from human, cattle, poultry, sheep, goat and water samples, respectively. Regardless of isolation source, C. jejuni and C. fetus were the most observed species with the prevalence rate of 17 (25.4%) each. C. coli was isolated only from 6 (9%) of samples, whereas, none of the tested samples were positive for C. lari. The use of stored water and practices of indoor and outdoor manure collecting, taking any specific protection while cleaning animal pen and washing hands before and after cooking were significantly associated with the human Campylobacter positivity. All (100%)of Campylobacter isolates from human, poultry, sheep, goat and water and 96% of isolates from cattle were resistant to at least one or more of the tested antimicrobials. More importantly, 95.5% the isolates were resistant to three or more classes of antimicrobials. Conclusion: The study found that multidrug resistant Campylobacter jejuni, coli, fetus and other Campylobacter with unidentified species were prevalent in the study area.Hence, considering the very significant effect of antimicrobial resistance, it is important for the country to have a national plan to advance the rational use of antimicrobials in the view of “One Health” approach.
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    Characterization of Group A Streptococci Isolated from Throat of Healthy School Children in Ethiopia
    (Addis Ababa University, 2005-07) Abdissa Alemseged; Asrat Daniel
    Post-streptococcal complications such as acute rheumatic fever (ARF) are reported to be common among Ethiopian children. Little is known, however, about the epidemiology of b-hemolytic streptococci, and Group A streptococci (GAS) in Ethiopia. A total of 937 randomly selected healthy schoolchildren residing in Addis Ababa (n=491), Gondar (n=265) and Dire-Dawa (n=181) were studied during a period November 2004 and January 2005. Throat specimens were collected and b-hemolytic streptococci were isolated on Columbia blood agar supplemented with Colistin-oxolonic acid. Gram reaction, catalase test Bacitracin susceptibility test and serological test were used to differentiate the organisms. In addition biochemical testing (API) was used to identify the species of isolates that could not be serogrouped. GAS were the predominant b-hemolytic streptococci with a proportion of 55% in Addis Ababa, 61% in Gondar, and 47% in Dire-Dawa of the serogroups. The carrier rate of GAS in healthy schoolchildren was 10.8% in Addis Ababa, 8.7% in Gondar and 9.4% in Dire- Dawa. Seven isolates that were nongroupable confirmed to be Streptococcus constellatus with API biochemical testing. Molecular (emm gene) typing was performed on 82 GAS isolates and 44 different emm types were detected. Among the 82 isolates 18 strains belonged to 13 new, previously undescribed emm subtypes. Known rheumatogenic strains accounted for 56% of isolates in Addis Ababa, 35% in Gondar and 0% in Dire-Dawa. Of the emm types prevalent in the study communities, 60% were not included in the 26 valent vaccine that was formulated for western countries. The distribution of emm types described in this study was compared with the study conducted on isolates collected 13 years ago in Addis Ababa, only 3/20 emm types identified in earlier study had been represented in the present study. Moreover, the type, which was predominant in the previous study, was not detected in the current one and the type that was predominant in our study was not detected in the previous one. x In conclusion, Group A was the most frequent serogroup among b hemolytic streptococci isolated from throat swabs of healthy schoolchildren in the selected three sites of Ethiopia and the carriage rate of GAS ranged from 8.7 to 10.8%. Tremendous diversity of emm types was observed in this first large scale emm type survey in Africa, and a significant proportion of them are known to be rheumatogenic. The association of the prevalent emm types not previously linked to rheumatic fever needs to be explored particularly in Dire-Dawa. Key Words: Beta-hemolytic streptococci, Group A Streptococci, Streptococcus pyogenes, M protein, emm typing
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    Determinants of Donor Fund Utilization: The Case of Public Sector Capacity Building Program (Pscap) Fund in Implementing Sectors of Oromia National Regional State
    (Addis Ababa University, 2015-10) Asrat Daniel; Ahmed Tufail
    The study was aimed to examine the determinants of donor funds utilization specifically PSCAP fund in implementing Sector of Oromia National Regional State. This study adopted a descriptive research design. Descriptive survey examines a situation as it is and yield quantitative information that can be summarized through statistical analyses. The target population was from Procurement, Finance and Property Administration and Internal Audit Departments of all PSCAP program implementing sectors. The sample of respondents was determined using stratified sampling which relies on mere chance to determine who would be selected in the sample and called for random selection in the inclusion of the cases into the sample. Stratification was done based on employees’ technical skills in both Departments of the five implementing sectors. A total of 64 employees out of a sample frame of 127 were selected from Procurement, Finance and Property Administration and Internal Audit Department of the implementing Sectors. This represents 50% of the total sample. Both primary and secondary methods of data collection were used for this study. Primary data was collected from questionnaires distributed to the Finance officers, Procurement Officers, Property Administration officers and Internal Audit officers. The secondary data was obtained from the records. The data was analyzed using Statistical Package for the Social Science (SPSS) version 20 to assist the researcher in answering the research questions. Descriptive and inferential statics were used for analysis of the research data. The descriptive statics part was analyzed using mean and standard deviation whereas correlation and regression were used from inferential statics. The data was summarized by use of descriptive statistics of the mean and standard deviation. Descriptive statistics aimed to summarize the data set while inferential statistics including correlation and regression analysis used the data collected to make conclusions on the whole population under study. The findings of the study established that there is positive and significant relationship between utilization of PSCAP fund and financial management and managerial factors (r=0.786 & 0.611, n=64, p=.000) as P<0.05 respectively. However, technical factors had negative, weak and insignificant relationship with PSCAP fund utilization(r= -0.091, n=64, p=0.476) as P > 005. The study also revealed that the three key factors: financial management, technical and managerial factors have significant effect on PSCAP fund utilization (β1, β2, β3= 0.647, -0.143, 0.348; p-value = 0.000, 0.036 & 0.000) which is less than α = 0.05) respectively. The study recommends that implementing sectors have to work towards improvements that will make the three determinants favorable to utilization of PSCAP fund. Key words: Financial management factors, Technical factors, Managerial factors
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    Epidemiology and Drug Resistance Pattern of Mycobacterium Tuberculosis in North West Ethiopia: Resource Limited Settings
    (Addis Ababa University, 2014-08) Airgecho Tekle; Woldeamanuel Yimtubezinash ; Asrat Daniel
    Background: Tuberculosis (TB) remains a major global health problem and ranks as the second leading cause of death among deaths caused by infectious diseases worldwide. Despite the availability of short course regimens as first line anti tuberculosis drugs, emergence of drug resistant Mycobacterium tuberculosis strains pose a major challenge to the prevention and control efforts of national tuberculosis programs (NTPs). In developing countries, including Ethiopia, the burden of tuberculosis and /or drug resistance profile of M.tuberculosis remains largely unexplored, mainly due to lack of quality controlled second line laboratory tests. Objectives: To assess the mycobacterial genotypic diversity and drug resistance pattern of M. tuberculosis complex in North West Ethiopia: resource limited settings. Moreover, it was to assess the knowledge, attitude and practice (KAP) of TB suspects towards tuberculosis Methods: Health care institution-based cross-sectional study was designed and conducted to assess: 1) Mycobacterial strain diversity 2) drug resistance pattern of Mycobacterium tuberculosis and 3) to evaluate the KAP of TB suspects towards TB in North West Ethiopia: resource limited settings. Study Subjects from health facilities in Benishangul Gumuz region and Awi zone of Amhara region who consented to participate were included in the study. 112 Smear positive samples were collected from study participants, stored and transported to Armauer Hansen Research Institute (AHRI) TB laboratory. At AHRI laboratory Lowenstein–Jensen (LJ) culture, drug susceptibility testing, RD9 deletion typing and Spoligotyping were performed from April 2013 through June 2014. Drug susceptibility testing (DST) was done by the proportion method on 24 well plates using 7H10 medium. The isolates were further characterized by spoligotyping and compared with updated SpolDB4.0 database to identify the circulating strains. Information for KAP study was collected on well-structured questionnaires. SPSS V.20 statistical software was used for data entry, data cleaning and analysis. Results: DST result of isolates from 87 samples showed one or more drug resistance in 16.5% with higher rate of monoresistance to isoniazid (6.9%). Multidrug resistance of 2(2.3%) was observed with 1(1.3%) and 1(8.3%) distribution among new and retreatment cases respectively. From 75 Mycobacterium tuberculosis isolates typed, 39 particular spoligotype patterns with 32(42.7%) of “New” isolates was observed. SIT289, SIT53, SIT149, SIT37 and SIT134 were the X predominant strains circulating in the study region with proportion of 14.7%, 12.0%, 8.0%, 5.3% and 4.0% respectively From 383 participants involved for KAP study 231(60.3%) were male and 152(39.7%) were female. Most frequent respondents’ sources of information about TB were health professionals in 131(34.2%) and 20(5.2%) had never heard about tuberculosis. 254(66.3%) knew that TB transmission is aerosol while 81(21.1%) do not know about it. 223(58.2%) responded TB transmission can be prevented by covering mouth and nose while coughing and sneezing but 125(32.6%) do not know about means of TB prevention. Respondents’ lack of knowledge 169(49.6%) and 63(18.5%) were reasoned accessibility and affordability for their delay in health service seeking behavior. Conclusion: Although current level of drug resistance appears to be low, the risk for a sudden increase is high because of the relatively higher mono resistance to INH. It is essential to maintain high quality DOTS in the area to keep MDR low. Majority of respondents also had poor knowledge and several misconceptions about TB that needs to be addressed through continuous health education on cause, transmission, prevention, treatment and services available. Key words: Mycobacterium tuberculosis, Strain diversity, Drug resistance, KAP, Benishangul Gumuz region, Northwest Ethiopia
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    Genomic Epidemiology and Molecular Features of Extended Spectrum β-Lactamase-Producing Enterobacteriaceae at Jimma Medical Center, Ethiopia.
    (Addis Ababa University, 2020-04) Sewunet Tsegaye; Asrat Daniel; Giske Christian ; Woldeamanuel Yimtubezinash; Aseffa Abraham
    Introduction: Infections caused by Enterobacterales are common causes of morbidity and mortality in hospitals and community. Several species of the Enterobacterales emerged with resistance property to extended/broad spectrum β-lactam antimicrobials. Such strains produce β-lactamase enzymes that can inactivate extended spectrum β-lactam antimicrobials, and hence, are called Extended Spectrum β-lactamase (ESBL)-producing strains. ESBL producing strains are increasingly being reported from different parts of the world revealing increased prevalence and shift of predominant molecular types over last two decades. However, there is a limited number of phenotypic studies in Ethiopia. The studies are not conclusive enough to unravel the magnitude, dissemination, and the actual burden of the problem. Objective: The study aimed to determine genomic epidemiology and molecular features of ESBL-producing Enterobacteriaceae at Jimma Medical Center (JMC), Ethiopia. Methods: A cross-sectional study was conducted at JMC from June-October 2016. Study participants were identified with the help of clinicians. And specimens relevant to clinical diagnosis of the patients were collected by experienced nurses. Specimens collected included: urine, stool, wound swab /aspirates, and sputum. Then specimens processed and inoculated on Mac Conkey agar, Blood agar and Salmonella Shigella agar at the microbiology laboratory and incubated at 37 o C for 18-24h. Species identification was performed primarily by conventional biochemical tests. Phenotypic screening and confirmation of ESBL-production was determined by Double Disc Synergy Test (DDST). Further validation/confirmation of species identification and antibiotic susceptibility test was performed by MALDI-TOF, and disk diffusion for ESBL-phenotype at Karolinska University Hospital, Department of Clinical microbiology, Sweden. Strains of E. coli and K. pneumoniae with ESBL phenotype were selected for whole genome sequencing (WGS) and sequenced on the Illumina (HiSeq 2500) sequencing platform at SciLifeLab, Stockholm, Sweden. Socio-demographic, clinical data and associated variables were collected using pre-designed structured questionnaire. Ethical clearance was obtained from relevant bodies. Study subjects were given information sheet about the study. Moreover, only study participants who provided written consent were recruited for the study. Descriptive data analysis was performed by using SPSS ver.20 statistical software. Sociodemographic, clinical data, and species related proportions were summarized by descriptive parameters. Where applicable, statistical association and significance was defined as p-value ≤ 0.05 as cut-off value for statistical association. Genome assembly and analysis was performed by using different bioinformatics tools, SPAdes version 3.9 was used for genome assembly. Enterobase, Center for Genomic Epidemiology (CGE), BIGsdb (Bacterial Isolate Genome sequence database), and Gubbins (Genealogies Unbiased By recomBination In Nucleotide Sequences), FastTree v.2, iTOL were tools used for genomic data analysis. Results: A total of 1,087 study participants either admitted to wards or seeking care as outpatient at medical, surgical, pediatric, and intensive care units at JMC were recruited. From these study participants, 58.9%(n= 640) were male and 41.1%(n=447) were female. A total of 1,087 specimens were collected, and 642 bacterial strains of different spp. were isolated. Most, 63.8% (n=255) were either E. coli(n=144) or Klebsiella spp.(n=111) and among nonfermenters, Pseudomonas aeruginosa and Acinetobacter spp. were predominant and accounted for 13% (n=87) of overall isolates. Within the four main units of the hospital, and among all specimens collected, E. coli and Klebsiella spp. were the most common strains isolated. The prevalence of ESBL-producing strains was (54.9%, 144/262) among E. coli and (76%, 111/146) among K. pneumoniae. A small proportion of E. coli strains (5.4%, 14/262) were non-susceptible to carbapenems. But only two E. coli strain encoded carbapenemaseng gene (bla NDM-1 and bla OXA-66 ). Similarly, a total of (22.6%, 33/146) of K. pneumoniae strains were non-susceptible to carbapenems, however, only one strain encoded bla NDM-1. Moreover, E. coli 96% (136/141) and K. pneumoniae 92%(102/111) were resistant to at least three classes of antimicrobials. Several genetic determinants of antimicrobial resistance were identified, where blaCTX-M-15 was the most prevalent ESBL, E. coli (88.4%, 123/139) and K. pneumoniae (84.4%, 92/109). The other genetic determinants included: bla TEM-1B (53.9%, 75/139) , blaOXA-1 (63.3%, 88/139) , dfrA17 (58.2%, 81/139), mph (67.2%, 94/139), aac(6’)-Ib-cr (62.6%, 87/139), and sul (68.3%,95/139) among E. coli strains, and bla TEM-1B (69.7%, 76/109), bla OXA-1 (34.8%, 38/109), sul (61.5%, 67/109), aac(6’)-Ib-cr (65.1%, 71/109), and dfrA27 (32.1%,35/109) among K.pneumoniae strains. Multi-locus sequesnce typing (MLST) revealed several sequence types (STs) carrying the ESBL genes. The most common sequence types (ST) encoding the blaCTX-M-15 were ST410(20/21), ST648 (15/15), ST131 (8/8), ST38 (6/6), ST44 (4/4) among E.coli. Among K. pneumoniae ST218 (7/7), ST15 (6/6), ST147 (6/6), ST17(6/6) and ST39 (5/5) had highest carriage of ESBL-genes. In both K. pneumoniae and E. coli strains, virulence determinant genes were invesitigated. EPEC, ExPEC/UPEC, EAEC were the prevalent pathotypes identified among E. coli strains. The O15/99:H30 ST38 (100%, n=6), O25:H4-ST131 (100%, n=8), O176:H34-ST130 (100%, n=5) encoded multiple toxins/virulence genes. On the other hand, K. pneumoniae strains were characterized by capsular and O-LPS antigens/types. Both classical and hypervirulent strains were identified as multi-drug resistant (MDR) strains carrying blaCTX-M-15 ESBL-genes. Plasmid replicon typing was performed, IncFII(pRSB107) (24.8%, n=32), IncQ1(20%, n=26)and Col(BS512) (14%, n=18) are the most commonly identified replicons among E. coli strains, and IncFIB( K/Mar/PQil) (32.9%, n=32), IncR (24.7%, n=24), and IncFII (14.4%, n=14) were prevalent replicons among K. pneumoniae strains. Single nucleotide polymorphysim (SNP)-based phylogenetic analysis of population structure showed multiple clusters of related strains. Though blaCTX-M-15 is the predominant ESBL-gene, other genetic determinants of resistance to b-lactams and other classes of antimicrobials were identified with a higher proportion in the same clonal complexes. Furthermore, some of these strains (both in E. coli and K. pneumoniae) were high-risk epidemic clones. The dissemination of bla CTX-M-15 mediated antimicrobial resistance was thus polyclonal. Some of the detected conal clusters suggest that an outbreak might have likely occured in the hospital. Furthermore, some possible risk factors for the increased prevalence of ESBL-strains were studied. Among these factors age less than five years, admission, pediatric unit and presence of underlying chronic illness showed statistically significant association with the higher prevalence of ESBL-strains. Gender and current use of antibiotics (reported use of antibiotics) showed no statistically significant association Conclusion: very high prevalence of ESBL among E. coli and K. pneumoniae was observed. The blaCTX-M-15 was the predominat ESBL-genetic variant. Most of the strains are MDR, also resistant to several other non-β-lactam antimicrobials. The detection of blaNDM-1 is a clear threat of evolving carbapenemases mediated carbapenem resistance. The spread of ESBLs is through multiple clusters of epidemic clones, and some of them are hypervirulent strains. The findings of this study are exteremly important primarly for the current hospital to look into it’s practice, to look into immediate preventive strategies including implemention of strict antimicrobial stewardship programmes and infection control strategies. The findings also shed light on genomic diversity and transmission dynamics of ESBL strains from Ethioipia and hence serve as a baseline data for researchers, planners and policy makers.
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    Magnitude of Extended Spectrum Beta-Lactamase Producing Multidrug Resistant Enterobacteriaceae Isolates from Patients Attending Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia
    (Addis Ababa University, 2020-02) Seman Aminu; Abebe Tamrat; Asrat Daniel
    Introduction: Multidrug resistant (MDR) bacteria constitute a global emergency. Of which extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-PE) have become the most common type. ESBL-PE are important causes of serious infections in the community as well as in hospital settings. Objective: To determine the magnitude and patterns of resistance of extended Spectrum βlactamase producing Enterobacteriaceae isolates from patients attending Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia. Methods: A cross sectional study was carried out to identify and characterize antimicrobial susceptibility pattern of Enterobacteriaceae isolates from TASH between July 2018 to November 2019. A total of 287 Enterobacteriaceae isolates were isolated from different specimens: blood, urine, wound site discharge, body fluids were analyzed using blood agar,MacConkey, Xylose lysine deoxycholate and salmonella shigella agar(SSA). Bacterial species identification was carried out using standard biochemical tests. Antimicrobial susceptibility testing and detection of ESBL production was performed using standard antibiotics on Muller Hinton agar using disk diffusion method and a combination disc method following 2018 CLSI guideline. Results: The overall magnitude of ESBL production was 62.7% (n=180/287). Female had higher proportion of ESBL positivity (66.1%, n=84/127) than male patients (60%, n=96/160). Among the Enterobacteriaceae isolates tested, 81 (72.3%) of the 112 K. pneumoniae, 21 (65.5%) of 32 K. oxytoca and 61 (58.7%) of 104 E.coli were ESBL producers. Comparable ESBL production by 158 (64.5 %) of 245 isolates from in patients and 22 (52.4%) of 42 isolates from outpatient was observed. There was no significant association between hospitalization and ESBL production (P=0.136, OR=0.606, 95 % CI). However, when we compare ESBL production of isolates from different wards a high ESBL production was noted among isolates from pediatrics ward 57(31.7%) of 180 isolates. The ESBL producing isolates showed highest and lowest level of resistance to ampicillin and amikacin 96.9% and 9.0 %, respectively. Two-hundred sixty four (91.9%) of the 287 isolates had MDR pattern. Conclusion: Significantly all isolates of Enterobacteriaceae showed higher rate of resistance to all tested antibiotics. But amikacin was found most effective for ESBL producers and no producers. High rate of ESBL and high numbers of MDR organisms were isolated among Enterobacteriaceae. Therefore, strengthening of monitoring antibiotics resistance in addition to regulation of the spread of resistance genes and preparing strong policies for monitoring utilization of antibiotics is required
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    Meningitis: Etiologic Agents and Antibiotic Susceptibility Pattern in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia
    (Addis Ababa University, 2009-12) Mengistu Mebratenesh; Mengistu Getahun ; Asrat Daniel; W/Amanuel Yimtubezinash
    Bacterial meningitis remains a common disease worldwide. Its most frequent causes are Nessieria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. In Ethiopia there are a number of previous published reports concerning meningitis. However most of the studies have been conducted during epidemic seasons. Meningitis is a serious emergency, which requires the early identification of the causative bacterium and its antibiogram. However, it is a usual practice to start empirical therapy before the completelaboratory result is available. Such practice requires knowledge of the most frequent etiological agents of meningitis and their antibiotic sensitivity patterns in the local population. This will help to influence the choice of initial therapy before a sensitivity result is available and in areas where laboratory facilities may not be available. To readdress this situation, this study was undertaken to isolate and identify the bacterial etiologic agents, including their antibiotic susceptibility pattern and serogroup/serotype isolated from patients with meningitis (n=340) visiting Tikur Anbessa University Hospital,Addis Ababa, Ethiopia, between November 2007 and June 2008. Of the patients investigated, 53.5% were males and 46.5% were females resulting in an overall male to female ratio of 1.2:1. Over half of the patients, 51.2% were children and the rest 32.6% and 16.2% were neonates and adults respectively. A total of 26 bacterial and fungal pathogens were isolated in all age groups. Of which, in neonates (n=2), children (n=16) and adults (n=8). Bacterial isolates accounted for 22 (84.6%) of the total isolates and the others 4 (15.4%) were fungal. In general, Streptococcus pneumoniae accounted for (36.4%) followed by Haemophilus influenzae type b and Nessieria meningitidis (Serogroup A and C)(13.6%) each. Cryptococcus neoformans was the only fungal isolate, which accounted for half of the isolates in adults.All gram positive bacteria were sensitive to Ceftriaxone, Ciprofloxacin, Chloramphenicol, Erythromycin and Rifampicin and Low level of resistance (<60%) were observed to Penicillin, Tetracycline and Trimethoprim sulphamethoxazole. exceptionally, all isolates of S.pneumoniae showed high level of resistance (>80%) to Gentamicin. Whereas, the gramnegatives showed high level of resistance (>80%) to Tetracycline and Trimethoprim sulphamethoxazole, intermediate level of resistance (60-80%) to Ampicilin and low level ofresistance (<60 %) to Ceftriaxone, Ciprofloxacin, Gentamycin, Chloramphenicol and Rifampicin. Most of the tested drugs were effective against gram-positive bacteria where as Gramnegative bacteria showed high level of resistance. Overall ceftriaxone and ciprofloxacin were the most effective drugs when compared to other drugs tested against the gram-positive and gram-negative bacteria. Multiple resistance (resistance to two or more drugs) was observed in 2/11(18.2%) and 11/11(100%) gram positive and gram-negative bacteria respectively. In conclusion, in the present study, S. pneumoniae was the predominant isolate followed by H. influenzae type b and N. meningitidis. The susceptibility pattern of isolates from the study showed that ceftriaxone and ciprofloxacin were the most effective drugs. Therefore, efforts should be directed towards early diagnosis and treatment of meningitis and continued reevaluation of the resistant patterns of organisms to optimize treatments and reduce complications. In addition, vaccination policies should reflect the primary etiologic agents, including pneumococcus and meningococcal serogroups A and C and H. influenzae type b.
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    Neonatal Sepsis: Bacterial Etiologic Agents and Their Antibiotic Susceptibility Pattern in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia
    (Addis Ababa University, 2008-10) Shitaye Demissie; Asrat Daniel ; W/Amanuel Yimtubezinash
    BACKGROUND: Neonatal sepsis is one of the most common reasons for admission to neonatal units in developing countries. It is also a major cause of mortality in both developed and developing countries. The type and pattern organisms that cause neonatal sepsis changes over time and vary from one hospital to another hospital, even in the same country. In addition the causative organisms have developed increased drug resistance for the last two decades. Maternal, neonatal and environmental risk factors have contributed for the development of sepsis. OBJECTIVES: This study was undertaken to determine the pattern of bacterial agents causing neonatal sepsis and to assess their susceptibility pattern to various antimicrobial agents in the Ethiopian setting. An attempt has been also made to identify the possible maternal and neonatal risk factors responsible for neonatal septicemia. METHODS: During the period of October 2006 and March 2007 a cross-sectional prospective study was conducted at the Department of Pediatrics and Child Health, Ethio- Swedish Children’s Hospital, Addis Ababa, Ethiopia. Blood cultures were performed from newborn babies (n=302, age: 0-28 days) admitted to the hospital with a clinical diagnosis of neonatal sepsis. Antimicrobial susceptibility testing was performed for all blood culture isolates according to the criteria of the National Committee for Clinical Laboratory Standards by disk diffusion method. RESULTS: Of the 302 patients, 46.4% were males and 53.6% were females (p >0.05) resulting in an overall male to female ratio of 0.8:1. The mean age of the neonates was 1.23 ± 8.96 days and 70.2% of neonates were below age of 1 day. A total of 298 (98.7%) neonates presented with early-onset sepsis (EOS) and four (1.3%) presented with late-onset sepsis (LOS). Out of the 302 neonates, 57.3% were preterm and 62.7% had low birth weight. The most prevalent clinical features of sepsis were hypothermia (84.8%), respiratory distress (72.8%), failure to feed (71.5%) and lethargy (30.1%). Among the 302 neonates admitted with suspected cases of sepsis, 59 (19.5%) had abnormal white blood cell count (high and low). Immature/total neutrophil ratio > 0.2 and <0.2 was observed in 58.0% and 42.0% peripheral blood smear examined respectively. Of the 302 neonates investigated for sepsis, 135 (44.7%) were positive for blood culture. The Gram-positive and negative bacteria accounted for 43.7% and 56.3% respectively (p> 0.05). The most common isolated organisms were Klebsiella spp. (39.2%) and Staphylococcus aureus (22.2%). Neonatal risk factors such as prematurity, low birth weight, abnormal WBC count (high and low) and I:T ratio > 0.2 were strongly associated with culture proven neonatal sepsis. No maternal risk factors were identified. In general both gram positive and negative bacteria isolated from blood culture showed low resistance rates (<60%) to ciprofloxacin, doxycycline, kanamycin, streptomycin, trimethoprim-sulphamethoxazole and tetracycline. Gram-negative bacteria showed high-level resistance (>80%) to ampicillin, ceftriaxone, cephalothin, chloramphenicol, and gentamicin. Ciprofloxacin was the most effective drug against the tested gram-positive and gram-negative bacteria. Multiple resistance (resistance to two or more drugs) was observed in 45.7% and 84.2% gram positive and gram negative bacteria respectively (p<0.05). CONCLUSION AND RECOMMENDATIONS: Klebsiella spp. and S. aureus were the most common organisms causing neonatal sepsis. Prematurity, low birth weight, abnormal WBC counts and I: T ratio > 0.2 were strongly associated with blood culture proven neonatal sepsis. Gram-negative bacteria showed high level of resistance to commonly used antibiotics. Ciprofloxacin was the most effective drug when compared to other drugs tested against the gram-positive and gram-negative bacteria. Multi-drug resistance detected in 67.4% isolates. The detection of multi-drug resistant isolates may further limit therapeutic options. Routine bacterial surveillance and the study of their resistance patterns must be an essential component of neonatal care. A knowledge of these patterns is essential when local polices on the uses of antibiotics are being devised. Key words: Neonatal sepsis, clinical features, bacterial pathogens, risk factors,antimicrobial susceptibility pattern, Ethiopia
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    Phenotypic and Molecular Characterizations of Salmonella Species in Ethiopia
    (Addis Ababa University Phenotypic and Molecular Salmonella Species, 2008-12) Beyene Getenet; Asrat Daniel; Mengistu Yohannes
    Background: Salmonella infections are very common and an important public health problem in many parts of the world. In sub-Saharan Africa there is very little direct data on strain type or antibiotic resistance. Research to date, as well as unpublished reports from different health institutions in Ethiopia, has indicated that salmonellosis is a common problem and the extensive use of the first line drugs has led to the development of multiple drug resistance at a level which could pose a serious problem in the near future. OBJECTIVES: The study was aimed at defining the serovars responsible for Salmonella infection, estimating the level of antibiotic resistance, investigate molecular basis of resistance and observing molecular polymorphism among Salmonella species isolated from children in Ethiopia. MATERIALS AND METHODS: Pediatric patients (n=1225) presenting with diarrhoea or fever from pediatric out patient department of Tikur Anbessa University Hospital, Addis Ababa (n= 825) and Jimma University Hospital, south west Ethiopia (n= 400) were investigated for enteric pathogens during January 2006 to June 2008. Forty eight Salmonella isolates collected in Tikur Anbessa hospital between January, 2004 and December, 2005 from similar age group were analyzed together. Stool specimens were collected for microscopic examination and culture. In addition blood specimens were obtained for culture. Brain Heart Infusion broth was used for blood culture and Selenite F broth enrichment followed by plating on Deoxycholate agar (DCA) and Xylose lysin deoxycholate agar (XLD) were used for stool culture. Identification of Salmonella species was carried out using API-20E, serology with antisera and strain were characterized using multilocus sequence typing (MLST). Based on resistance pattern, site of collection and type of specimen in which the isolate was detected, S. Concord isolates were selected and investigated for plasmid profile, incompatibility grouping, pulsed field gel electrophoresis (PFGE), MLST profile and fliC gene sequencing using standard procedures. In addition, 48 previously collected Salmonella isolates before the commencement of the present study were sero-grouped and serotyped. RESULTS: A total of 463 entropathogens were isolated from 1225 pediatric patients. The isolates were: 65 Salmonella species, 61 Shigella species and 337 parasites. Among the 113 xi Salmonella isolates (65 + 48 previously collected), serogroup C, B, D and E were isolated at a frequency of 78.8%, 11.5%, 8% and 1.8% respectively. Most of the Salmonella isolates were from stool (68%) and the rest were from blood (32%). No isolate was detected from both blood and stool of the same patient. A total of 12 serotypes were identified namely; S. Concord (82), S. Colindale (1), S. Gatow (3), S.Laronchelle (1), S. Garoli (1), S. Colorado (1), S. Typhimurium (7) S. Paratyphi B(2), S. Haifa (1) S. Typhi (2) S. Enteritidis (4), and S. Butantan (2). Eighty nine percent of the group C isolates were S. Concord. S. Concord isolates were highly resistant to ampicillin, trimethoprimsulfamethoxazole,ceftriaxone, amoxicillin,chloramphenicol, gentamicin, and tetracycline. Low resistance rate was observed for nalidixic acid and ofloxacin and there was no resistance to ciprofloxacin by disk diffusion test. However, E-test result indicated the presence of one resistant, one intermediate and twenty two (26.8%) Concord isolates which showed reduced susceptibility to ciprofloxacin. The extended spectrum beta lactamase (ESBL) screening test result showed that 98.8% of S. Concord were positive for ESBL production. Plasmid analysis showed that all characterized S. Concord isolates harbored multiple copies of small and large plasmids. The molecular weight of plasmids varied from less than five to 170 kb with 120, 118 and 95 kb being the most prevalent. Plasmid replicons A/C, I1 and incFI were found in the majority of the isolates. Different plasmid studies indicated that A/C and incFI replicons are associated with multi drug resistance (MDR) Salmonella isolates. A total of 16 pulse field gel electrophoresis (PFGE) profiles were seen among the 23 Concord group and 5 non Concord isolates. Every sequence type (ST) had a unique PFGE profile which indicates that S. Concord in Ethiopia is in an endemic situation, rather than a spread of a clonal type (has many different point sources/reservoirs). The same ST and PFGE types were found in Addis Ababa and in Jimma suggesting movement of infected people/reservoirs between the two cities. Multilocus sequence typing (MLST)analysis showed the presence of a total of seven STs among 58 isolates. ST533, ST534 and ST599 are single locus variants. Because they differ in only one of the seven loci, they are closely related genetically and make a single Concord group. Distantly related serotypes like Gatow and Colindale had sequence types that differ by more than one allele from Concord group. Molecular serotyping using sequencing of the fliC gene was able to differentiate xii between S. Haifa and the S. Concord group of isolates indicating the possibility of its role in molecular serology. For our strain collection, it seems therefore that fliC sequencing can complement MLST for classification and strain differentiation. CONCLUSION AND RECOMMENDATIONS: Salmonellosis in children in these two regions of Ethiopia is mainly due to non-typhoidal salmonellae particularly with S. Concord. This is different from other countries where S. Enteritidis and S. Typhimurium accounted for the majority of salmonellosis. Both phenotypic and genotypic characterization indicated that S. Concord is highly resistant and present in endemic situation in Ethiopia. The presence of many multi-drug resistant strains containing genes for ESBL production and the emergence of reduced susceptibility to ciprofloxacin in this study posed a major concern in the search for efficient antimicrobial therapy of Salmonella infections in the near future. Therefore more comprehensive studies should be designed to trace its source and distribution within the country and to monitor antibiotic resistance pattern over time.
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    Phenotypic and Molecular Characterizations of Staphylococcus Aureus Isolates from Human and Dairy Cows in Mekele, Northern Ethiopia
    (Addis Ababa University, 2019-03) Abrha Alem ; Asrat Daniel ; Woldeamanuel Yimtubezinash ; Wang Shu; Gebreyes Wondwossen
    Introduction: Staphylococcus aureus is one of the common colonizers of human and several animal species with the capacity to cause serious diseases. It is one of the global public health priority pathogens due to its ubiquitous nature, ability to cause life-threatening diseases and continuous evolvement of drug resistance. The emergence and spread of multidrug resistant strains, mainly MRSA, initially in the health care setting, later in the community and recently in the veterinary side with a capacity of transmission to human is of special concern. Several molecular techniques have been used successfully for reliable identification, determination of specific virulence and drug resistance genes and to understand spread and distribution of specific S. aureus strains. In Ethiopia, despite several reports on phenotypic studies of S. aureus isolates from human and animals, data on molecular characterizations is scarce. Objectives: To isolate S. aureus from human skin and soft tissue infections, milk of dairy cows and nares of dairy farm workers in Mekele, Northern Ethiopia; and then to determine the antimicrobial resistance pattern, virulence genes profile and genotypic diversity and relatedness of the isolates. Methods: A cross sectional study was conducted during period of March 2016 to August 2017 G.C. A total of 811 non-duplicate specimens (355 wound/pus swabs from human patients with skin and soft tissue infections [SSTIs] attending Ayder referral hospital, 71 nasal swabs from dairy farmers and 384 milk samples from dairy cows) were collected to isolates S. aureus. Culture and identification of S. aureus was performed using standard laboratory procedures. Antimicrobial susceptibility testing was performed following theCLSI guideline. All phenotypic characterizations of S. aureus were done at the Microbiology laboratory of Ayder Referral Hospital, Mekele, Ethiopia. AlSaureus isolates were stored at −700C and shipped to USA for Molecular characterizations at the Infectious Diseases Molecular Epidemiology Laboratory (IDEML) of the Ohio State University and the Public Health Research Institute of the State University of New Jersey, USA. Molecular characterizations include confirmation of S. aureus by nuc detection, mecA /mecC detection, toxin genes profiling (pvl, nine se genes and tsst-1), spa typing and SCCmec typing. Results: From the 811 samples, 193 (23.8%) were positive for S. aureus using phenotypic and molecular methods; where 123 (63.7%) were from human SSTIs, 22 (11.4%) from nares of dairy farmers and 48 (24.9%) from milk of Dairy cows. Overall, high percentage of resistance was observed for Penicillin (92.7%); however, no resistance was found for Vancomycin, Daptomycin and Rifampin. Four (2.1%) of the 193 isolates were MRSA where all were from human and were both cefoxitin resistant and mecA positive. Two of them carried SCCmec type III and the other two SCCmec type IV. Moreover, 26% of S. aureus isolates were multidrug resistant. Regarding toxin genes carriage, 66.8% of the total isolates possessed at least one of the 11 targeted toxin genes. Of the 11 genes, pvl was the most frequently detected (carried by 36.8%) followed by seg and sei (29% each), seb (19.2%), sea(16%) and sec (15%). Interestingly, pvl was found significantly higher (p<0.001) on isolates from SSTI (53.7%) compared to nasal (13.6%) and cows’ milk (4.2%) isolates. In addition,tsst-1 was found in human isolates only. The spa typing yielded 61 different types among the 190 successfully typed isolates. Out of these, five were novel spa types, namely spa typet17828, t17829, t17830, t17831 and t17832. The most dominant spa type among the SSTI isolates was t355 (19%) followed by t306 (8.3%); whereas in milk spa type t042 (61.7%) was the dominant one followed by t2856 (19.1%). The present study also noted that spa type t042, t306, t085, t2856 and t5338 were detected from both human and dairy cows. Based on the spa sequence information, 90.5% of the total S. aureus were assigned in to 12 probable clonal complexes (CCs), namely CC1, CC5, CC8, CC15, CC22, CC25, CC80, CC81, CC88, CC121, CC152 and CC239. The leading CC among the human clinical isolates was CC152 (20.7%) followed by CC15 (19%) and CC5 (16.5%). However, most of the S. aureus isolates from dairy cows (83%) were CC80 followed by CC5 (6.4%). Conclusion: In the present study, S. aureus was isolated from skin and soft tissue infections, nares of farm workers and milk of dairy cows. Highest resistance was observed against penicillin. No resistance was observed against Vancomycin, Daptomycin and Rifampin. Multidrug resistance was observed in a quarter of the isolates. mecA positive S. aureus (MRSA) strains were of human origin. Majority of S. auresus isolates possessed at least 1 of the 11 toxin genes. The spa typing of S. aureus resulted in 61 different types and showed diversity. Based on the spa sequence information, S. aureus isolates were assigned into 12 probable clonal complexes (CCs). Among these CCs, CC1, CC5, CC15 and CC80 were shared between human and bovine isolates. Findings of the present study indicate that S. aureus is a significant cause of skin and soft tissue infections in human and intramammary infections in dairy cows. Hence, measures to reduce/prevent infection such as improving infection prevention practices in the hospital, keeping hygiene of the dairy cows and educating the community to practice the best possible personal hygiene are recommended. Further study should be conducted in different regions of the country to have conclusive data for policy makers, clinicians and researchers.
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    Prevalence and Antimicrobial Resistance of Bacterial Isolates With Special Emphasis on Enterobacteriaceae Among Children Suspected for Septicemia and Urinary Tract Infection in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia
    (Addis Ababa University, 2014-05) Hailu Melese; Asrat Daniel
    Background: Blood stream and urinary tract infections are a major cause of mortality and morbidity ofthe pediatric population. Extended spectrum beta-lactamase (ESBL) producing and carbapenemresistant enterobacteriaceae is the challenge for controlling now days. Assessing the prevalent bacteriaand their antibiotic resistance helps to provide effective therapies, develop rational prescriptionprograms and make policy decisions. Objective: To determine the prevalence and antimicrobial resistance of bacterial isolates with specialemphasis on enterobacteriaceae among children suspected for septicemia and urinary tract infection inTikur Anbessa University Hospital, Addis Ababa, Ethiopia. Method: A cross sectional study was conducted from January 10 to March 30/2014 at Tikur AnbessaUniversity Hospital, Addis Ababa, Ethiopia. A total of 322 study participants who were suspected forsepticemia and/or UTI were recruited. All blood and urine samples were cultured on Blood andMacConkey agar. All culture positives were characterized by colony morphology, biochemical tests using the standard procedure. Significant bacteriuria was determined for all culturepositive urine samples. Antimicrobial susceptibility test was performed for all bacterial isolates usingKirby-Bauer method. ESBL was detected using combination disk & double disk synergy methods onMuller Hinton agar. Carbapenemase were detected by Modified Hodge method using Meropenem. Alldemographic & laboratory data were entered to EPI INFO & exported to SPSS version 20 for analysis. Result: The overall prevalence of bacteria isolates from blood and urine cultures was 17.1%. From 177blood samples 13.0% (n=23/177) and from 145 urine samples 22.1% (n=32/145) were culture positives.Coagulase negative Staphylococci & Klebsiella ozaenae were the predominate bacteria isolated inblood and urine cultures respectively. Most of them 89.1% (n=49/55) developed multidrug resistance(MDR≥2 drugs) to most commonly used antibiotics. Multiple resistances were observed in 71.42% ofGram positive and 95.11% Gram negative isolates. Prevalence of ESBL producing and carbapenem resistant enterobacteriaceae was 78.57% and 12.12% respectively. Conclusion: The choice of drugs in the treatment of bacteria isolates from blood and urine is quitenarrow today due to the wide scale resistance to common antibiotics. The emergence of MDR calls forcontinuous monitoring & reviewing of antimicrobial policy in hospitals and the country at large. Key terms: Septicemia, UTI, Bacterial isolates, antimicrobial resistance pattern, ESBL, Carbapenemresistance, Tikur Anbessa University Hospital, Addis Ababa, Ethiopia
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    Prevalence of Bacterial Vaginosis Among Pregnant Women Attending Antenatal Care in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia.
    (Addis Ababa University, 2012-06) Mengistie Zemenu; Woldeamanuel Yimtubezinas; Asrat Daniel
    Background: Bacterial vaginosis is one of the most common genital tract infections among reproductive age group. It is associated with different gynecologic and poor obstetric out come. The prevalence of bacterial vaginosis varies from country to country even in the same country it varies at population of interest. Different social and sexual factors can contribute for development of bacterial vaginosis. Bacterial vaginosis can be easily diagnosed by combination of two Amsel’s criteria. Objectives: This study was undertaken to determine the prevalence of bacterial vaginosis and to evaluate the accuracy of Amsel’s criteria individually or in combination of two for the clinical diagnosis of bacterial vaginosis among pregnant women attending Antenatal care in Tikur Anbessa University Hospital, Addis Ababa, Ethiopia. An attempt has also been made to identify the possible risk factors associated with bacterial vaginosis. Methods: In this observational study during the period from November 2011 to April 2012, 252 pregnant women were screened for bacterial vaginosis in Tikur Anbessa University Hospital. Vaginal swabs were collected for pH determination, saline wet mount microscopic examination to detect clue cells, KOH preparation for whiff test and Gram-stain evaluation of vaginal flora for diagnosis of bacterial vaginosis by Nugent scoring system. Accuracy of clinical diagnosis using individual and two of Amsel’s criteria was evaluated. Results: The prevalence of bacterial vaginosis was 18.3% by Amsel’s two of three criteria and 19.4% using Gram stain Nugent scoring system. The prevalence of bacterial vaginosis was 31.6% and 15.9% among symptomatic and asymptomatic pregnant women respectively. A high percentage of bacterial vaginosis positive pregnant women were asymptomatic (63.3%) whereas the remaining 36.7% bacterial vaginosis positive pregnant women were complaining abnormal vaginal discharge with or without unpleasant smell. Comparing with Gram stain Nugent scoring methods the clinical diagnosis by Amsel’s criteria had sensitivity of 85.7%, specificity of 98%, and positive predictive value of 91.3% and negative predictive value of 96.6%. The most sensitive and specific individual criterion was clue cells and with highest positive and negative predictive value. Amsel’s criteria with the lowest sensitivity and specificity were whiff test and vaginal pH respectively. Combination of clue cells with vaginal pH test were the highest in sensitive while whiff test with clue cells were the highest in specificity than the other combined two Amsel’s criteria. After adjusted for other factors multiple life time sexual partner (OR: 8.6; 95% CI: 2.5, 29) and previous history of spontaneous abortion (OR: 5.9; 95% CI: 1.5, 23) had remained significantly associated with prevalence of bacterial vaginosis. They were the most likely to be a risk factors for bacterial vaginosis infection. Conclusion and Recommendation: The prevalence of bacterial vaginosis among pregnant women was higher in asymptomatic pregnant women and associated with the factors multiple lifetime sexual partner and previous spontaneous abortion. Amsel’s criteria diagnosis of bacterial vaginosis can be simplified using a combination of the two criteria, vaginal pH and clue cells, in settings where time or Gram staining is not available. So using these simplified diagnostic criteria is better to screen pregnant women as a routine activityduring antenatal care.
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    Prevalence of Group B Streptococcus Colonization among Pregnant Women Attending Antenatal clinic of Hawassa Health Center, Hawassa, Ethiopia
    (Addis Ababa University, 2008) Mohammed Musa; Asrat Daniel
    Background: Group B streptococcus (GBS) or Streptococcus agalactiae are members of the normal flora of the female genital tract. During labor GBS may infect the newborn, leading to neonatal sepsis and meningitis. GBS emerged in the 1970s as one of the most frequent causes of sepsis and meningitis in neonates and young infants. Since the mid-1960s, GBS has become the major cause of bacterial infections in the perinatal period, including bacteraemia, amnionitis, endometritis, and urinary tract infection in pregnant women. Rates of GBS colonization vary widely throughout the world due to differences in laboratory investigation methods, regional variations and racial differences. Report on prevalence of GBS among pregnant women in Ethiopia is very limited. Objectives: The objective of this study was to determine the prevalence of group B Streptococcal (GBS) colonization among pregnant women attending antenatal clinic at Hawassa Health centre, Hawassa Ethiopia and analyze risk factors related to GBS colonization. Methods: A total of 139 pregnant women were screened for GBS colonization between May and June 2010. Standard microbiological methods were used to isolate and identify GBS from vaginal and anorectal swabs obtained from study subjects. Antimicrobial susceptibility test was performed for all GBS isolates according to the criteria of the National Committee for Clinical Laboratory Standards by disk diffusion method. Results: A total of 29 out of 139 pregnant women studied (20.8%) were colonized by GBS. No statistically significant association was observed for GBS colonization with any of sociodemographic characteristics of the study subjects including age, occupation, type of contraceptive used, types of gravida, number of antenatal clinic visits etc. All GBS strains were susceptible to penicillin, ampicillin, vancomycin and gentamicin, Low level of resistance (<60%) were observed against erythromycin, tetracycline, ceftriaxone, chloramphenicol, ciprofloxacin and norfloxacin. Conclusion and recommendations: this study including 139 pregnant women, confirmed the prevalence of GBS colonization to be around 21%. This prevalence was compared with findings reported from developed and developing countries and have reached comparable level. However, further epidemiological investigations should be conducted in different parts of the country in order to know the actual GBS colonization rate in pregnant women and consider implementation of prevention plans using intrapartum antibiotics prophylaxis to prevent early onset GBS-neonatal diseases. Key words: Group B Streptococcus (GBS), risk factors, Antibiotic susceptibility, Ethiopia.
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    Serological and Molecular Characterization of Hepatitis B, C and D Viruses Infections Among Health Professionals in Ras Desta and Tikur Anbessa Hospitals, Addis Ababa, Ethiopia
    (Addis Ababa University, 2008) Yimer Kedir; Asrat Daniel
    The risks for occupational infection with blood-borne pathogens (HBV, HCV, and HIV) have been a source of concern among health professionals because of their frequent and often substantial exposures to patient blood and body fluids and all of which are associated with significant morbidity and mortality. A cross sectional study was conducted in order to assess the prevalence of HBV, HCV, and HDV infections among HCW’s (n=267) and risk factors associated with these viral infections. After each volunteer study subject has completed a standardized questionnaire, 10 ml of venous blood was collected from each individual for serological and molecular processing. ELISA method was used to detect HBs and HBe antigens, antibodies to HBc, HBs, HDV and HCV in the serum samples. Molecular methods were used to detect HBV DNA and HCV RNA. One hundred and eight (40.4%) of the health professionals use gloves consistently, 221 (82.3%) had at least a one time history of cutaneous exposure to blood ungloved, 165 (61.8%) of health care workers had at least a one time history of cutaneous exposure to any of the body fluids, 162 (60.6%) had splashed blood or body fluids in their faces, 158 (59.2 %) had sustained needle stick injury and 83 (31.1%) sustained sharp injury. The overall prevalence of HBV infection was found to be 51.3% (95% CI: 45-57%) and the infection prevalence increases with age (p<0.01). HBsAg was detected in 9.7 % (95% CI: 6-13.2%) of HCW’s. Absence of vaccination was the only risk factor associated with hepatitis B infection(p=0.001). Though only a single individual had HBeAg (4.5%; 95% CI: 2-7%), HBV DNA was detected by PCR in serum sample from 17 of the 24 subjects (71%). The hepatitis B vaccination rate was found to be 13%. Ninety three percent of individuals who were vaccinated with a single dose of HBs vaccine had achieved protective antibody. ‘Anti-HBc only’ positive cases were detected in 6 (2.2%; 95% CI: 0.004-3.96%) individuals however HBV DNA was not detected in any of the samples tested. HCV infection was detected in 1 (0.37%) study subject. None of the HBsAg positive individuals had marker of HDV infection. Our data suggest that occupational exposures to blood and body fluids occur frequently with low universal precaution practice and vaccination coverage. Hepatitis B infection is widespread and characterized by high level of immunity through natural infection and the presence of continuous horizontal transmission.Absence of vaccination is the only risk factor associated with hepatitis B infection. In this study HCV and HDV infection prevalence is very low. Serologic response rate to a single dose of viii hepatitis B vaccine was much higher than experiences from other countries. HBeAg negative serological status and low viral replication levels characterize chronic hepatitis B virus-infected health professional. ‘Anti-HBc only’ is found in the health professionals but none of the subjects had detectable HBV DNA. This suggests the need to do future study in ‘Anti-HBc only’ case.