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.

Shiga toxin-producing Escherichia coli O157:H7 among diarrheic patients and their cattle in Amhara National Regional State, Ethiopia.

BACKGROUND: Shiga toxin-producing Escherichia coli O157:H7 (STEC O157:H7) is a zoonotic pathogen that causes diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome worldwide. This study aimed to determine the prevalence, antibiotic susceptibility, and associated risk factors of STEC O157:H7 among diarrheic patients and their cattle. METHODS: A cross-sectional study was conducted among diarrheic patients and their cattle in Amhara National Regional State, Ethiopia from December- 2020 to June- 2022. A total of 1,149 diarrheic patients and 229 cattle were included in the study. STEC O157:H7 detection was done using culture, latex agglutination test, and polymerase chain reaction on diarrheic stool samples and recto-anal mucosal swabs of cattle. Antibiotic susceptibility tests were performed using disk diffusion techniques. Risk factors association were identified using binary and multivariable logistic regression analysis. RESULTS: The overall prevalence of STEC O157:H7 in diarrheic patients and their cattle was 11.1% (128/1149) and 14.4% (33/229) respectively. High percentage of the study subjects were found in under-five children (34.5%). Age less than 5 (AOR: 4.02, 95%CI:1.608-10.058,P = 0.003), and greater than 64 years old (AOR:3.36, 95% CI:1.254-8.986, P = 0.016), presence of diarrheic patient in the house (AOR:2.11, 95%CI:1.309-3.390, P = 0.002), availability of cattle in the house (AOR:2.52, 95%CI:1.261-5.049, P = 0.009), and habit of consuming raw foods (AOR:4.35, 95%CI:2.645-7.148, P = 0.000) were risk factors. Antibiotic resistance was shown in 109(85.2%), and 31(93.9%) isolates from diarrheic patients and their cattle respectively. The highest levels of antibiotic resistance were found to tetracycline (54.7%, 69.7%) in diarrheic patients and their cattle respectively. Multiple drug resistance was also observed among 56(43.8%) and 11(33.3%) isolates in diarrheic patients and their cattle respectively. CONCLUSION: Our study showed high prevalence of STEC O157:H7 in diarrheic patients and their cattle. Therefore, health education should be given to the community on how to care for animals, proper sanitation, and the impact of raw food consumption.

Evaluation of the diagnostic performance of EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit in Amhara National Regional State, Ethiopia: A multi-center cross-sectional study.

Rotavirus is the leading cause of morbidity and mortality due to acute gastroenteritis among children under five years globally. Early diagnosis of rotavirus infection minimizes its spread and helps to determine the appropriate management of diarrhea. The aim of this study was to evaluate the performance of EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit for the diagnosis of rotavirus infection among diarrheic children under five years in Ethiopian healthcare settings. A total of 537 children with diarrhea were enrolled from three referral hospitals in Amhara National Regional State, Ethiopia. The samples were tested using one-step RT-PCR and EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit (KTR-917, Epitope Diagnostics, San Diego USA) in parallel. Diagnostic performance of the rapid test kit was evaluated using the one-step RT-PCR as a gold standard. The sensitivity, specificity, and predictive values of the rapid test kit were determined. Moreover, the agreement of the rapid test kit with one step RT-PCR was determined by kappa statistics and receiver operators' curve (ROC) analysis was done to assess the overall diagnostic accuracy of the rapid test kit. Fecal Rotavirus Antigen Rapid Test Kit has shown a sensitivity of 75.5% and specificity of 98.2%. The kit was also found to have 89.9% and 95.0% positive and negative predictive values, respectively. The Fecal Rotavirus Antigen Rapid Test Kit has shown a substantial agreement (78.7%, p = 0.0001) with one-step RT-PCR. The overall accuracy of the Fecal Rotavirus Antigen Rapid Test Kit was excellent with the area under the ROC curve of 86.9% (95% CI = 81.6, 92.1%) (p = .0001). Thus, Fecal Rotavirus Antigen Rapid Test is a sensitive, specific, user-friendly, rapid, and equipment-free option to be used at points of care in Ethiopian health care settings where resource is limited precluding the use of one step RT-PCR. Furthermore, the kit could be used in the evaluation and monitoring of rotavirus vaccine effectiveness in the aforementioned settings.

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.

The influence of anti-cancer therapies on lymphocyte subpopulations of lung cancer patients.

Introduction: There are limited data on the influence of different anti-cancer therapies on lymphocyte subpopulations and their relationships to survival of non-small cell lung cancer (NSCLC) patients. This study aimed to assess the effect of immunotherapy, chemotherapy, immunochemotherapy, adjuvant chemotherapy after surgery, and antibodies against Vascular Endothelial Growth Factors (VEGF) on B cell, T cell, and NK cell subpopulations, and the survival time of NSCLC patients. Methods: A total of 32 consecutive NSCLC patients were recruited at Pulmonology Clinic, Leipzig from January 2018 to March 2020 and enrolled in this study. Immunophenotyping was done using a FACS Canto II flow cytometer (BD Biosciences) before the administration of the planned therapy and during therapy with up to 7 observational windows for each patient targeting 130 immunologic parameters. Results: Absolute transitional B cells was significantly increased after immunotherapy (p = 0.032), immunochemotherapy (p = 0.030), and antibodies against VEGF (p = 0.024). Similarly, absolute counts and percentage of B cells were significantly increased after adjuvant chemotherapy (p = 0.023). However, absolute counts and percentage of transitional B cells are significantly decreased after chemotherapy (p = 0.001). Activated cytotoxic T cells were significantly increased after immunotherapy (p = 0.031) and immunochemotherapy (p = 0.030). The overall survival rate of NSCLC patients was 31%. Conclusions: In conclusion, this study suggests that different types of anti-cancer therapies affect lymphocyte subpopulations of NSCLC patients. Further large-scale and multicentre studies are required to confirm our results and to evaluate the prognostic value of lymphocyte subpopulations.

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.

High proportion of RR-TB and mutations conferring RR outside of the RRDR of the rpoB gene detected in GeneXpert MTB/RIF assay positive pulmonary tuberculosis cases, in Addis Ababa, Ethiopia.

BACKGROUND: Globally, TB is the leading cause of infectious disease morbidity and mortality with many diagnostic uncertainties. Access to affordable and rapid diagnostics remained a major challenge for many developing countries which bear the greatest burden of TB delaying the initiation time to treatment. OBJECTIVE: This study aimed to assess the GeneXpert MTBRIF assay probe utility for the detection of pulmonary TB and Rifampicin-resistant TB cases in Addis Ababa, Ethiopia. MATERIALS AND METHODS: A cross-sectional study was performed from October 2019 to July 2020 in Saint Peter TB Specialized Hospital in Addis Ababa metropolitan area, Ethiopia. This study enrolled 216 clinically suspected new presumptive pulmonary TB cases confirmed by GeneXpert MTB/RIF Assay. Sociodemographic and clinical characteristics were captured using a structured tool. Data were entered in Microsoft Excel 2019, checked for inconsistency, cleaned promptly, and exported to IBM SPSS Statistics for Windows, Version 26.0. Armonk, N.Y: IBM Corp, the USA for analysis. Descriptive analysis and binary and multivariate logistics regression were performed and all statistical significance was determined at a 95% confidence level. RESULTS: The majority of the study participants, 55.1% [119/216] were males aged 6-80 years. The prevalence of RR MTB was 11.11% [24/216]. A higher proportion of RR TB was found in female patients [54.2%, 13/24], in patients in the age group of 30-50 years [45.8%, 11/24], in married individuals [62.5%, 15/24], in persons whose residence is urban [79.2%, 19/24], in persons who had a previous history of TB symptoms [100%, 24/24], in persons who had a history of contact with active and LTBI [33.3%, 8/24], and in persons who had a history of HIV and IDUs [41.7%, 10/24]. Occupation (AOR 22.868, 95% CI 1.655-316.022, p = 0.019), history of previous PTB+ (AOR 4.222, 95% CI 1.020-17.47, p = 0.047), and history of HIV and IDUs (AOR 4.733, 95% CI 1.416-15.819, p = 0.012) were independent predictors associated with RR-TB emergence. The commonest mutation 62.5% [15/24] was found in probe E (codons 529-533) region. There was no mutation associated with probe A (codons 507-511), probe B (codons 511-518), and probe C (codons 518-523) regions, as well as no combination of missed probes, was revealed. However, 12.5% [3/24] of RR TB patients were found without unidentified missed probe types detected outside of the RRDR. The delta Ct max was >4.0 and the highest proportion of 35.6% [77/216] RR TB was detected in samples of medium DNA load. CONCLUSION: The proportion of RR-TB we observed in this study was high. Similarly, a higher proportion of RR TB was detected outside of the RRDR. Moreover, a significant number of the GeneXpert MTB/RIF Assay probes were identified as unhybridized and this critical observation would mean that most of the probes had no or minimal utility in this geographical region. This calls for further studies to uncover mutation in the rpoB gene conferring RR and reshape TB triage and definite diagnostic algorithm in Ethiopia.

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.

Flow-Cytometry Intracellular Detection and Quantification of HIV1 p24 Antigen and Immunocheckpoint Molecules in T Cells among HIV/AIDS Patients.

Introduction: HIV p24 antigen-positive T cells measured by flow cytometry (FCM) correlate directly with HIV viral load, inversely with CD4 + T cells, and decrease with antiretroviral therapy (ART). However, the sensitivity of FCM assays depends on the protocol of intracellular staining. Therefore, this study aimed to evaluate the diagnostic performance of our FCM protocol for detection of HIV p24-positive T cells and measure the level of immunocheckpoint molecules (PD1 and TIM3) in T cells. Methods: The study was conducted at the University of Leipzig hospital between January 2020 and November 2020. Viremic and ART-suppressed HIV-positive patients and negative controls were included in this study. HIV1 p24 KC57-, p24 28B7-, PD1-, and TIM3-positive CD4 and CD3 T cells were analyzed from whole blood using a BD FACS Canto II flow cytometer equipped with FACSDiva software. HIV1 p24 antigen FCM results were compared with HIV1 RNA viral load results measured by Alinity M assays on the fully automated random-access platform. We analyzed the data using SPSS 20. Results: The absolute CD4 + and CD4 +:CD8 + T-cells ratio showed a significant inverse correlation with HIV1 viral load. Moreover, the absolute CD4+ T-cells count showed a significant inverse correlation with p24 KC57-positive CD4 T cells. The percentage of p24 KC57, p24 28B7, and double-positive CD4 T cells showed significant correlation with HIV1 viral load. PD1 expressing CD4 T cells were higher in ART-viremic cases than controls, while TIM3-expressing CD4 T cells were lower in ART-viremic cases than controls. Sensitivity, specificity, PPV, and NPV of p24 KC57-positive CD4 T cells were 64%, 82%, 78%, and 69%, respectively, for the diagnosis of HIV infection and 55%, 73%, 40%, and 83%, respectively, for treatment monitoring. Conclusion: Our protocol showed moderate performance for the diagnosis of HIV infection and treatment monitoring. Therefore, the p24 KC57 but not the p24 28B7 clone could be considered as a simple alternative method for rapid diagnosis of HIV infections and treatment monitoring, particularly in low- and middle-income countries.

Prediction of drug resistance by Sanger sequencing of Mycobacterium tuberculosis complex strains isolated from multidrug resistant tuberculosis suspect patients in Ethiopia.

BACKGROUND: Ethiopia is one of the high multidrug-resistant tuberculosis (MDR-TB) burden countries. However, phenotypic drug susceptibility testing can take several weeks due to the slow growth of Mycobacterium tuberculosis complex (MTBC) strains. In this study, we assessed the performance of a Sanger sequencing approach to predict resistance against five anti-tuberculosis drugs and the pattern of resistance mediating mutations. METHODS: We enrolled 226 MTBC culture-positive MDR-TB suspects and collected sputum specimens and socio-demographic and TB related data from each suspect between June 2015 and December 2016 in Addis Ababa, Ethiopia. Phenotypic drug susceptibility testing (pDST) for rifampicin, isoniazid, pyrazinamide, ethambutol, and streptomycin using BACTEC MGIT 960 was compared with the results of a Sanger sequencing analysis of seven resistance determining regions in the genes rpoB, katG, fabG-inhA, pncA, embB, rpsL, and rrs. RESULT: DNA isolation for Sanger sequencing was successfully extracted from 92.5% (209/226) of the MTBC positive cultures, and the remaining 7.5% (17/226) strains were excluded from the final analysis. Based on pDST results, drug resistance proportions were as follows: isoniazid: 109/209 (52.2%), streptomycin: 93/209 (44.5%), rifampicin: 88/209 (42.1%), ethambutol: 74/209 (35.4%), and pyrazinamide: 69/209 (33.0%). Resistance against isoniazid was mainly mediated by the mutation katG S315T (97/209, 46.4%) and resistance against rifampicin by rpoB S531L (58/209, 27.8%). The dominating resistance-conferring mutations for ethambutol, streptomycin, and pyrazinamide affected codon 306 in embB (48/209, 21.1%), codon 88 in rpsL (43/209, 20.6%), and codon 65 in pncA (19/209, 9.1%), respectively. We observed a high agreement between phenotypic and genotypic DST, such as 89.9% (at 95% confidence interval [CI], 84.2%-95.8%) for isoniazid, 95.5% (95% CI, 91.2%-99.8%) for rifampicin, 98.6% (95% CI, 95.9-100%) for ethambutol, 91.3% (95% CI, 84.6-98.1%) for pyrazinamide and 57.0% (95% CI, 46.9%-67.1%) for streptomycin. CONCLUSION: We detected canonical mutations implicated in resistance to rifampicin, isoniazid, pyrazinamide, ethambutol, and streptomycin. High agreement with phenotypic DST results for all drugs renders Sanger sequencing promising to be performed as a complementary measure to routine phenotypic DST in Ethiopia. Sanger sequencing directly from sputum may accelerate accurate clinical decision-making in the future.

Origin and Global Expansion of Mycobacterium tuberculosis Complex Lineage 3.

Mycobacterium tuberculosis complex (MTBC) Lineage 3 (L3) strains are abundant in world regions with the highest tuberculosis burden. To investigate the population structure and the global diversity of this major lineage, we analyzed a dataset comprising 2682 L3 strains from 38 countries over 5 continents, by employing 24-loci mycobacterial interspersed repetitive unit-variable number of tandem repeats genotyping (MIRU-VNTR) and drug susceptibility testing. We further combined whole-genome sequencing (WGS) and phylogeographic analysis for 373 strains representing the global L3 genetic diversity. Ancestral state reconstruction confirmed that the origin of L3 strains is located in Southern Asia and further revealed multiple independent introduction events into North-East and East Africa. This study provides a systematic understanding of the global diversity of L3 strains and reports phylogenetic variations that could inform clinical trials which evaluate the effectivity of new drugs/regimens or vaccine candidates.

Effects of Intermittent Hypoxia in Training Regimes and in Obstructive Sleep Apnea on Aging Biomarkers and Age-Related Diseases: A Systematic Review.

Several studies have assessed the effects of intermittent hypoxia-normoxia training (IHNT), intermittent hypoxia-hyperoxia training (IHHT), and obstructive sleep apnea (OSA) on aging and age-related diseases in humans; however, the results remain contradictory. Therefore, this review aims to systematically summarize the available studies on the effects of IHNT, IHHT, and OSA on aging and age-related diseases. Relevant studies were searched from PubMed, Google Scholar, Cochrane Library databases, and through manual searching from reference lists of eligible studies. A total of 38 eligible studies were included in this systematic review. IHHT and IHNT provide positive effects on several age-related parameters including quality of life, cognitive and physical functions, plasma level of glucose and cholesterol/LDL, systolic blood pressure, red blood cells, and inflammation. Moreover, moderate intermittent hypoxia induces telomerase reverse transcriptase (TERT) activity and telomere stabilization, delays induction of senescence-associated markers expression and senescence-associated ß-galactosidase, upregulates pluripotent marker (Oct4), activates a metabolic shift, and raises resistance to pro-apoptotic stimuli. On the contrary, intermittent hypoxia in OSA causes hypertension, metabolic syndrome, vascular function impairment, quality of life and cognitive scores reduction, advanced brain aging, increase in insulin resistance, plasma hydrogen peroxide, GSH, IL-6, hsCRP, leptin, and leukocyte telomere shortening. Thus, it can be speculated that the main factor that determines the direction of the intermittent hypoxia action is the intensity and duration of exposure. There is no direct study to prove that IHNT/IHHT actually increases life expectancy in humans. Therefore, further study is needed to investigate the actual effect of IHNT/IHHT on aging in humans. Systematic Review Registration: www.crd.york.ac.uk/prospero, identifier CRD42022298499.

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.

Bacterial profile, antimicrobial susceptibility patterns, and associated factors of community-acquired pneumonia among adult patients in Gondar, Northwest Ethiopia: A cross-sectional study.

INTRODUCTION: Community-acquired pneumonia is associated with higher morbidity, hospitalization, and mortality in adults. Likewise, antimicrobial resistance has increased in recent decades in Ethiopia. Therefore, this study was aimed to determine the bacterial isolates, their antimicrobial susceptibility patterns, and factors associated with community-acquired pneumonia among adult patients in Gondar, Northwest Ethiopia. MATERIALS AND METHODS: This institutional-based cross-sectional study was conducted from April to June 2021. Sociodemographic, clinical, and other relevant data were collected using a pre-tested questionnaire. A total of 312 sputum specimens were collected using sputum cups and inoculated into blood agar, chocolate agar, mannitol salt agar, and MacConkey agar plates, which were then incubated at 37°C for 24 hours. The bacterial isolates were identified based on Gram staining, colony characteristics, and biochemical tests. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Inducible clindamycin resistance among the S. aureus isolates was detected by the D-test. Data were entered using EPI data version 4.6 and analyzed using SPSS version 20. P-value ≤ 0.05 at 95% CI was considered statistically significant. RESULTS: Of 312 cases, 39.4% (n = 123; 95% CI: 34.1%-44.9%) were found to have culture-confirmed pneumonia. The most common isolates were K. pneumoniae (31.0%, n = 39), S. pneumoniae (26.2%, n = 33), and S. aureus (20.6%, n = 26). The gram-positive bacteria were susceptible to chloramphenicol (100%) and clindamycin (96.6%). Gram-negative bacteria were susceptible to gentamicin (87.5%), azithromycin (87.1%), ciprofloxacin (86.6%), and ceftriaxone (79.0%) but highly resistant to ampicillin (100%), followed by tetracycline (87.1%), doxycycline (86.4%), co-trimoxazole (80.6%), and amoxicillin-clavulanic acid (79.0%). Overall, 72.2% of the isolates were multi-drug resistant to K. pneumoniae (94.9%, n = 37), E. coli (93.8%, n = 15), and S. pneumoniae (72.7%, n = 24). Only, 7.7% of S. aureus isolates showed inducible clindamycin resistance. Aging (AOR: 3.248, 95% CI: 1.001-10.545, p = 0.050), a history of pneumonia (AOR: 7.004, 95% CI: 3.591-13.658, p = 0.001), alcohol use (AOR: 6.614, 95% CI: 3.399-12.872, p < 0.001), and overcrowded living conditions (AOR: 4.348, 95% CI: 1.964-9.624, p = 0.001) were significantly associated with culture-positive sputum. CONCLUSION AND RECOMMENDATIONS: This study found a high prevalence of bacteria-caused community-acquired pneumonia among adults and low susceptibility to ampicillin, tetracyclines, and amoxicillin-clavulanic acid. Therefore, culture-based bacterial identification and local antibiotic susceptibility testing should be performed regularly. Additionally, new insights into vaccine coverage against highly multi-drug resistant bacteria, particularly K. pneumoniae, are necessary.

Bacterial Profile, Antimicrobial Susceptibility Pattern, and Associated Risk Factors Among Patients With Wound Infections at Debre Markos Referral Hospital, Northwest, Ethiopia.

Wound infections are associated with morbidity and mortality in developing countries. Thus, this study aimed to assess bacterial profile, antimicrobial susceptibility pattern and risk factors among wound infection suspected patients. A hospital-based cross-sectional study was conducted on 201 participants at Debre Markos referral hospital from January to May 2019. Sociodemographic data were collected using pre-designed questionnaire and swabs from different types of wounds were collected, and inoculated onto mannitol salt agar, blood and MacConkey agar plate for isolation and identification. Antimicrobial susceptibility tests were done using modified Kirby-Bauer disk diffusion technique. Out of 201 swabs analyzed, 72.6% were culture positive and 10.9% of them had co-infections, and 162 bacterial isolates obtained from 146 swab samples. Staphylococcus aureus was the most frequently isolated which accounted for 32.1% of isolates followed by Pseudomonas aeruginosa, 15.4%. The susceptibility patterns of ciprofloxacin, gentamycin and ceftriaxone were 77.8%, 69.1%, and 68.5%, respectively. The MDR rate of gram positive and gram negative isolates were 69.7% and 82.3%, respectively. Anatomically located wounds near a site of potential contamination, inadequate management of moisture, exudate or edema, mechanism of wound production, presence of predisposing condition and being urban in residence significantly associated with wound infections. S. aureus and P. aeruginosa were the predominant causes of wound infections. Ciprofloxacin, gentamycin and ceftriaxone were the most effective antimicrobials. Periodic surveillance of isolates involved in wound infection and their antimicrobial susceptibility is recommended for effective management of patients.

Seroprevalence and Associated Factors of Hepatitis B and C Virus Among Pulmonary Tuberculosis Patients Attending Health Facilities in Gondar Town, Northwest Ethiopia.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) are hepatotropic viruses whose primary replication occurs in the liver. Despite the significant clinical importance of early screening of hepatitis B and C virus infection in decreasing the hepatotoxicity effect of anti-tuberculosis drugs, screening of hepatitis B and C virus among tuberculosis (TB) patients before treatment has not been practiced in Ethiopia. Thus, this study was conducted to determine the seroprevalence and associated factors of HBV and HCV infections among pulmonary TB (PTB) patients attending health facilities in Gondar, Northwest Ethiopia. METHODS: A cross-sectional study was conducted among 145 bacteriologically confirmed PTB patients from January 1 to May 30, 2019. After obtaining a signed informed consent from each participant, data on socio-demographic, clinical, and associated factors were collected using a structured pre-tested questionnaire. Besides, a blood sample was collected to determine HBsAg and HCV antibodies by enzyme linked immune sorbent assay (ELISA). The data were entered and analyzed using SPSS version 21. A Fisher's exact test was used to see the relationship between dependent and independent variables, and a p-value ≤0.05 was considered as statistically significant. RESULTS: Out of the 145 PTB patients screened, 5 (3.4%) patients tested positive for HBsAg, yet none of them were found to be positive for anti-HCV. Besides, the proportion of HIV-positive was 12 (8.3%). History of hospital admission (P= 0.005), tattooing (P= 0.009) and dental extraction (P=0.003) were significantly associated with HBsAg. CONCLUSION: Although anti-HCV antibodies were not detected, the prevalence of HBV was relatively high in tuberculosis patients. This study highlights the need for the introduction of routine screening of viral hepatitis markers for all TB patients before anti-TB treatment for better management of patients. Likewise, further clinical and epidemiological studies are needed.

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.

Malaria Infection is High at Transit and Destination Phases Among Seasonal Migrant Workers in Development Corridors of Northwest Ethiopia: A Repeated Cross-Sectional Study.

PURPOSE: Malaria is a leading public health problem in Ethiopia. Every year, thousands of seasonal farm workers travel to farm corridors in Northwest Ethiopia and fall at risk of malaria infection. However, the magnitude of malaria infection and risk factors during harvest time were not well identified. This study aimed at estimating the prevalence and risk factors of malaria infection among seasonal migrant workers in Northwest Ethiopia. METHODS: A repeated cross-sectional study was conducted at transit and destination phases in Metema district from September 15, 2018 to October 30, 2019. Data were collected using a structured questionnaire. A capillary blood sample was collected to examine infection with malaria parasite using a microscope. A multivariate logistic regression technique was used to determine risk factors. RESULTS: The malaria prevalence at transit and destination phases among migrant workers was 13.5% (95% CI: 12.07-14.93%) and 18.7% (95% CI: 16.40-21.02%), respectively. The combined prevalence was 16.1% (95% CI: 14.67-17.63%). The odds of malaria infection among migrant workers at the destination phase was 1.5 (OR=1.5, 95% CI 1.167-1.846) times higher compared to the transit phase. Education (AOR=8.198; 95% CI: 4.318-15.564), knowledge of antimalarial drugs (AOR=2.4; 95% CI: 1.43-3.95), and use of long-lasting insecticidal nets (AOR=5.0; 95% CI: 3.34-4.43) were significantly associated with malaria infection at migration phases. CONCLUSION: This study showed that the burden of malaria among seasonal migrant workers was high at transit and destination phases. Malaria prevalence was higher at the destination phase compared to the transit phase. A tailored malaria prevention intervention is needed including awareness creation, screening, treatment, repellent, and prophylaxis at both phases to reduce malaria infections.

Pertussis among patients with clinically compatible illness in the Amhara Regional State, Ethiopia.

BACKGROUND: Pertussis is an acute respiratory tract disease caused by Bordetella pertussis. In 2014, 24.1 million pertussis cases, resulting in 160,700 deaths, were estimated to have occurred worldwide. This study aimed to determine the epidemiology of pertussis among patients with clinically compatible illness who visited selected hospitals in the Amhara Regional State of Ethiopia. METHODS: A cross-sectional study design was used to review pertussis patients with clinically compatible illness. Nasopharyngeal swabs were collected from 515 patients from July 2018 through February 2019. DNA was extracted from all nasopharyngeal swabs and samples were analyzed using real-time (RT-) PCR. Crude and adjusted odds ratios with corresponding 95% confidence intervals were estimated using bivariable and multivariable logistic regression analysis, respectively. RESULTS: The overall prevalence of Bordetella species among the study participants was 156 of 515 (30.3%) [95% CI = 26.4-34.6] as determined by Bordetella RT-PCR, including: 65 (41.7%) B. pertussis, 89 (57.1%) indeterminate B. pertussis, one (0.6%) Bordetella holmesii and one (0.6%) Bordetella parapertussis. CONCLUSIONS: This study found that pertussis is potentially endemic and a common health problem among patients visiting health institutions in the Amhara Regional State of Ethiopia. More data regarding pertussis in Ethiopia could inform development of effective prevention strategies.

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.