Effectiveness of bubble continuous positive airway pressure for treatment of children aged 1-59 months with severe pneumonia and hypoxaemia in Ethiopia: a pragmatic cluster-randomised controlled trial.

BACKGROUND: The safety and efficacy of bubble continuous positive airway pressure (bCPAP) for treatment of childhood severe pneumonia outside tertiary care hospitals is uncertain. We did a cluster-randomised effectiveness trial of locally made bCPAP compared with WHO-recommended low-flow oxygen therapy in children with severe pneumonia and hypoxaemia in general hospitals in Ethiopia. METHODS: This open, cluster-randomised trial was done in 12 general (secondary) hospitals in Ethiopia. We randomly assigned six hospitals to bCPAP as first-line respiratory support for children aged 1-59 months who presented with severe pneumonia and hypoxaemia and six hospitals to standard low-flow oxygen therapy. Cluster (hospital) randomisation was stratified by availability of mechanical ventilation. All children received treatment in paediatric wards (in a dedicated corner in front of a nursing station) with a similar level of facilities (equipment for oxygen therapy and medications) and staffing (overall, one nurse per six patients and one general practitioner per 18 patients) in all hospitals. All children received additional care according to WHO guidelines, supervised by paediatricians and general practitioners. The primary outcome was treatment failure (defined as any of the following: peripheral oxygen saturation <85% at any time after at least 1 h of intervention plus signs of respiratory distress; indication for mechanical ventilation; death during hospital stay or within 72 h of leaving hospital against medical advice; or leaving hospital against medical advice during intervention). The analysis included all children enrolled in the trial. We performed both unadjusted and adjusted analyses of the primary outcome, with the latter adjusted for the stratification variable and for the design effect of cluster randomisation, as well as selected potentially confounding variables, including age. We calculated effectiveness as the relative risk (RR) of the outcomes in the bCPAP group versus low-flow oxygen group. This trial is registered with ClinicalTrial.gov, NCT03870243, and is completed. FINDINGS: From June 8, 2021, to July 27, 2022, 1240 children were enrolled (620 in hospitals allocated to bCPAP and 620 in hospitals allocated to low-flow oxygen). Cluster sizes ranged from 103 to 104 children. Five (0·8%) of 620 children in the bCPAP group had treatment failure compared with 21 (3·4%) of 620 children in the low-flow oxygen group (unadjusted RR 0·24, 95% CI 0·09-0·63, p=0·0015; adjusted RR 0·24, 0·07-0·87, p=0·030). Six children died during hospital stay, all of whom were in the low-flow oxygen group (p=0·031). No serious adverse events were attributable to bCPAP. INTERPRETATION: In Ethiopian general hospitals, introduction of locally made bCPAP, supervised by general practitioners and paediatricians, was associated with reduced risk of treatment failure and in-hospital mortality in children with severe pneumonia and hypoxaemia compared with use of standard low-flow oxygen therapy. Implementation research is required in higher mortality settings to consolidate our findings. FUNDING: SIDA Sweden and Grand Challenges Ethiopia.

Effectiveness of Bubble Continuous Positive Airway Pressure (BCPAP) for Treatment of Children Aged 1-59 Months with Severe Pneumonia and Hypoxemia in Ethiopia: A Pragmatic Cluster Randomized Controlled Clinical Trial.

Despite the beneficial effect of bubble continuous positive airway pressure (BCPAP) oxygen therapy for children with severe pneumonia under the supervision of physicians that has been shown in different studies, effectiveness trials in developing country settings where low-flow oxygen therapy is the standard of care are still needed. Thus, the aim of this study is to assess the effectiveness of bubble CPAP oxygen therapy compared to the WHO standard low-flow oxygen therapy among children hospitalized with severe pneumonia and hypoxemia in Ethiopia. This is a cluster randomized controlled trial where six district hospitals are randomized to BCPAP and six to standard WHO low-flow oxygen therapy. The total sample size is 620 per arm. Currently, recruitment of the patients is still ongoing where the management and follow-up of the enrolled patients are performed by general physicians and nurses under the supervision of pediatricians. The primary outcome is treatment failure and main secondary outcome is death. We anticipate to complete enrollment by September 2022 and data analysis followed by manuscript writing by December 2022. Findings will also be disseminated in December 2022. Our study will provide data on the effectiveness of BCPAP in treating childhood severe pneumonia and hypoxemia in a real-world setting.

Active case detection and treatment of malaria in pregnancy using LAMP technology (LAMPREG): a pragmatic randomised diagnostic outcomes trial-study protocol.

INTRODUCTION: Malaria is one of the major public health problems in sub-Saharan Africa. It contributes significantly to maternal and fetal morbidity and mortality in affected countries. This study aims to evaluate the impact of enhanced case detection using molecular testing called loop-mediated isothermal amplification (LAMP) on birth outcomes in a prospective study design. METHODS AND ANALYSIS: A pragmatic randomised diagnostic outcomes trial will be conducted in several health institutes in different Ethiopian regions. Women (n=2583) in their first and second trimesters of pregnancy will be included in the study and individually randomised to the standard of care or enhanced case detection arms, and followed until delivery. Enrolment will encompass the malaria peak transmission seasons. In the standard of care arm, a venous blood sample will be collected for malaria diagnosis only in symptomatic patients. In contrast, in the intervention arm, mothers will be tested by a commercially available Conformité Européene (CE)-approved LAMP malaria test, microscopy and rapid diagnostic test for malaria regardless of their symptoms at each antenatal care visit. The primary outcome of the study is to measure birth weight. ETHICS AND DISSEMINATION: The study was approved by the following ethical research boards: Armauer Hansen Research Institute/ALERT Ethics Review Committee (FORM AF-10-015.1, Protocol number PO/05/20), the Ethiopia Ministry of Science and Higher Education National Research Ethics Review Committee (approval SRA/11.7/7115/20), the Ethiopia Food and Drug Administration (approval 02/25/33/I), UCalgary Conjoint Health Research Ethics Board (REB21-0234). The study results will be shared with the institutions and stakeholders such as the Ethiopia Ministry of Health, the Foundation for Innovative Diagnostics, WHO's Multilateral initiative on Malaria - Tropical Diseases Research (TDR-MIM), Roll Back Malaria and the Malaria in Pregnancy Consortium. The study results will also be published in peer-reviewed journals and presented at international conferences. TRIAL REGISTRATION NUMBER: NCT03754322.

A Single Dose of ChAdOx1 nCoV-19 Vaccine Elicits High Antibody Responses in Individuals with Prior SARS-CoV-2 Infection Comparable to That of Two-Dose-Vaccinated, SARS-CoV-2-Infection-Naïve Individuals: A Longitudinal Study in Ethiopian Health Workers.

Single-dose COVID-19 vaccines, mostly mRNA-based vaccines, are shown to induce robust antibody responses in individuals who were previously infected with SARS-CoV-2, suggesting the sufficiency of a single dose for those individuals in countries with limited vaccine supply. However, these important data are limited to developed nations. We conducted a prospective longitudinal study among Ethiopian healthcare workers who received a ChAdOx1 nCoV-19 vaccine. We compared the geometric mean titers (GMTs) of the SARS-CoV-2 receptor-binding domain (RBD)-specific IgG antibodies in 39 SARS-CoV-2 naïve participants and 24 participants previously infected with SARS-CoV-2 (P.I.), who received two doses of ChAdOx1 nCoV-19 vaccine across the two post-vaccination time points (at 8 to 12 weeks post single dose and two dose vaccinations). We noted that the GMT (1632.16) in naïve participants at 8-12 weeks post first dose were comparable to the GMT (1674.94) observed in P.I. participants prior to vaccination. Interestingly, P.I. participants had significantly higher antibody titers compared to naïve participants, after both the first (GMT, 4913.50 vs. 1632.16) and second doses (GMT, 9804.60 vs. 6607.30). Taken together, our findings show that a single ChAdOx1 nCoV-19 dose in previously SARS-CoV-2 infected individuals elicits similar, if not higher, antibody responses to those of two-dose-vaccinated naïve individuals.

A single dose ChAdOx1 nCoV-19 vaccine elicits high antibody responses in individuals with prior SARS-CoV-2 infection comparable to that of double dose vaccinated SARS-CoV-2 infection naïve individuals.

Background A single dose COVID-19 vaccines, mostly mRNA-based vaccines, are shown to induce robust antibody responses in individuals who were previously infected with SARS-CoV-2, suggesting the sufficiency of a single dose to those individuals. However, these important data are limited to developed nations and lacking in resource-limited countries, like Ethiopia. Methods We compared receptor-binding domain (RBD)-specific IgG antibodies in 40 SARS-CoV-2 naïve participants and 25 participants previously infected with SARS-CoV-2, who received two doses of ChAdOx1 nCoV-19 vaccine. We measured the antibody response in post-vaccination blood samples from both groups of participants collected at four different post-vaccination time points: 8- and 12-weeks after each dose of the vaccine administration using an in-house developed ELISA. Results We observed a high level of anti-RBD IgG antibodies titers 8-weeks after a single dose administration (16/27; 59.3%) among naïve participants, albeit dropped significantly (p<0.05) two months later, suggesting the protective immunity elicited by the first dose ChAdOx1 nCoV-19 vaccine will likely last for a minimum of three months. However, as expected, a significant (p<0.001) increase in the level of anti-RBD IgG antibodies titers was observed after the second dose administration in all naïve participants. By contrast, the ChAdOx1 nCoV-19 vaccine-induced anti-RBD IgG antibody titers produced by the P.I participants at 8- to 12-weeks post-single dose vaccination were found to be similar to the antibody titers seen after a two-dose vaccination course among infection- naïve participants and showed no significant (p>0.05) increment following the second dose administration. Conclusion Taken together, our findings show that a single ChAdOx1 nCoV-19 dose in previously SARS-CoV-2 infected individuals elicits similar antibody responses to that of double dose vaccinated naïve individuals. Age and sex were not associated with the level of vaccine-elicited immune responses in both individuals with and without prior SARS-CoV-2 infection. Further studies are required to assess the need for a booster dose to extend the duration and amplitude of the specific protective immune response in Ethiopia settings, especially following the Omicron pandemic.

Plasmodium falciparum histidine-rich protein 2 and 3 genes deletion in global settings (2010-2021): a systematic review and meta-analysis.

BACKGROUND: The usefulness of histidine-rich protein-2/3 (HRP2/3)-based rapid diagnostic tests of malaria due to Plasmodium falciparum has been threatened by the appearance of mutant PfHRP2/3 genes. This study was undertaken to determine the global pooled estimates of PfHRP2/3gene deletions. METHODS: Relevant publications were identified from electronic databases such as; PubMed, EMBASE, and MEDLINE online. Besides, all the relevant literatures were retrieved through Google and Google Scholar. STATA software was used for data analysis. The pooled estimates were calculated using random effect model. The summary estimates were presented using forest plots and tables. RESULTS: A total of 27 studies were included in the systematic review. However, only 24 and 17 studies were included for PfHRP2 and 3 gene deletion meta-analysis, respectively. The prevalence of PfHRP2 gene deletion across the individual studies ranged from the highest 100% to the lowest 0%. However, the meta-analysis result showed that the global pooled prevalence of PfHRP2 and PfHRP3 gene deletions were 21.30% and 34.50%, respectively. The pooled proportion of PfHRP2 gene deletion among false negative PfHRP2-based RDTs results was found to be 41.10%. The gene deletion status was higher in South America and followed by Africa. The pooled estimate of PfHRP2 gene deletion among studies, which did not follow the WHO PfHRP2/3 gene deletion analysis protocol was higher than their counter parts (21.3% vs 10.5%). CONCLUSIONS: This review showed that there is a high pooled prevalence of PfHRP2/3 gene deletions in Plasmodium falciparum confirmed isolates and also a high proportion of their deletions among false-negative malaria cases using PfHRP2-based RDT results. Hence, malaria diagnosis based on PfHRP2-based rapid tests seems to be less sensitive and warrants further evaluation of PfHRP2/3 gene deletions.

Whole-Genome Sequences of SARS-CoV-2 Isolates from Ethiopian Patients.

Three complete severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes from Ethiopian patients were compared with deposited global genomes. Two genomes belonged to genetic group 20A/B.1/GH, and the other belonged to genetic group 20A/B.1.480/GH. Enhancing genomic capacity is important to investigate the transmission and to monitor the evolution and mutational patterns of SARS-CoV-2 in this country.

Prevalence, Infection Intensity and Associated Factors of Soil-Transmitted Helminthiasis Among School-Aged Children from Selected Districts in Northwest Ethiopia.

BACKGROUND: Globally, soil-transmitted helminths affect beyond a billion people and cause 1.9 million disability-adjusted life years worldwide. It affects children disproportionately due to their unaware activities like walking barefoot, playing with dirty objects that might be contaminated with feces. The control of soil-transmitted helminths principally relies on periodic deworming using either a single dose of albendazole/mebendazole. To assure the effectiveness of this measure, performing continuous parasitological survey is necessary. Herein, the prevalence, intensity and associated factors of soil-transmitted helminth infections were assessed among school-aged children in northwest Ethiopia. METHODS: A cross-sectional study design was conducted among school-aged children (6-14 years old) from January 21st to February 21st/2019. Multistage sampling technique was employed. A Kato-Katz concentration technique was utilized to detect STHs in stool samples. Moreover, risk factors for STH infections were assessed using well-structured questionnaire. Bivariate and multivariate analyses were used to assess the association between explanatory and the outcome variables. The magnitude of the association was measured using the adjusted odds ratio (AOR) and 95% confidence interval (CI). A P-value <0.05 was considered statistically significant. RESULTS: The overall STHs prevalence in this study was 32.3% (95% CI: 29-35.6%) with Ascaris lumbricoides being the predominant species (24.3%) followed by hookworm (8.9%) and Trichuris trichiura (1%). Most (80.3%) of the infected school-aged children had light-intensity infections. Age of 11 years and above (AOR, 12.9, 95% CI, 1.6-103.6, P=0.004), being residing in Chuahit district (AOR, 3.9, 95% CI, 2.3-6.5, P<0.001), and untreated water supply (AOR, 1.7, 95% CI, 1.1-2.7, P=0.018) were identified as predictors for the overall STH prevalence. CONCLUSION: Our findings revealed STH infections are considerable health problems in the study areas. Thus, public health interventions such as provision of safe water supply, health education, and de-worming programs should be regularly implemented in the study areas.

The challenges of COVID-19 testing in Africa: the Ethiopian experience.

Novel coronavirus disease (COVID-19) is spreading rapidly and creating a huge economic, social and public health challenge worldwide. Although currently an effective vaccine is ready, its distribution is limited, and hence the only currently available lever to reduce transmission is to identify and isolate individuals who are contagious. Thus, testing for SARS CoV-2 has a paramount importance. However, testing in many African countries including Ethiopia has multidimensional growing challenges. Here, we tried to identify, categorize and summarize the challenges of COVID-19 testing in Africa from Ethiopian experience.

Evaluation of sample pooling for screening of SARS CoV-2.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has revealed the global public health importance of robust diagnostic testing. To overcome the challenge of nucleic acid (NA) extraction and testing kit availability, an efficient method is urgently needed. OBJECTIVES: To establish an efficient, time and resource-saving and cost-effective methods, and to propose an ad hoc pooling approach for mass screening of SARS-CoV-2. METHODS: We evaluated pooling approach on both direct clinical and NA samples. The standard reverse transcriptase polymerase chain reaction (RT-PCR) test of the SARS CoV-2 was employed targeting the nucleocapsid (N) and open reading frame (ORF1ab) genomic region of the virus. The experimental pools were created using SARS CoV-2 positive clinical samples and extracted RNA spiked with up to 9 negative samples. For the direct clinical samples viral NA was extracted from each pool to a final extraction volume of 200µL, and subsequently both samples tested using the SARS CoV-2 RT-PCR assay. RESULTS: We found that a single positive sample can be amplified and detected in pools of up to 7 samples depending on the cycle threshold (Ct) value of the original sample, corresponding to high, and low SARS CoV-2 viral copies per reaction. However, to minimize false negativity of the assay with pooling strategies and with unknown false negativity rate of the assay under validation, we recommend pooling of 4/5 in 1 using the standard protocols of the assay, reagents and equipment. The predictive algorithm indicated a pooling ratio of 5 in 1 was expected to retain accuracy of the test irrespective of the Ct value samples spiked, and result in a 137% increase in testing efficiency. CONCLUSIONS: The approaches showed its concept in easily customized and resource-saving manner and would allow expanding of current screening capacities and enable the expansion of detection in the community. We recommend clinical sample pooling of 4 or 5 in 1. However, we don't advise pooling of clinical samples when disease prevalence is greater than 7%; particularly when sample size is large.

Treatment efficacy and re-infection rates of soil-transmitted helminths following mebendazole treatment in schoolchildren, Northwest Ethiopia.

BACKGROUND: Transmission of soil-transmitted helminth (STH) infection remains high in Ethiopia. This study aimed at assessing the therapeutic efficacy of mebendazole against soil-transmitted helminths and determining the re-infection rates of the parasites among schoolchildren in Northwest Ethiopia. METHODS: A school-based cross-sectional study was conducted. Data was collected using a structured questionnaire. Stool specimens were examined using direct wet mount microscopy and Kato-Katz methods. Schoolchildren who tested positive for soil-transmitted helminths were treated with 500 mg single-dose of mebendazole. Cure and egg reduction rates were evaluated 2 to 3 weeks post treatment. Moreover, the re-infection rate of these parasites among those who were cured was determined 1 year after treatment. Data were analyzed using SPSS version 20. P value < 0.05 was considered as statistically significant. RESULT: A drug efficacy study was conducted on 62, 52, and 14 children infected by Ascaris lumbricoides (A. lumbricoides), hookworm, and Trichuris trichiura (T. trichiura), respectively. The cure rates (CR) of mebendazole against A. lumbricoides, hookworm, and T. trichiura were found to be 96.9%, 23.1%, and, 28.6%, respectively. The egg reduction rate (ERR) of A. lumbricoides was found to be 99.6% whereas 49.6% and 56.3% were reported for hookworm and T. trichiura, respectively. Eighty schoolchildren who were treated and cured from any STH infections were included for the determination of re-infection rate. Out of 80 children, 36.3% (29/80) were found to be re-infected after 1 year: 22 (75.9%), 6 (20.7%), and 1 (1.3%) of study participants were re-infected with A. lumbricoides, hookworm, and both infections, respectively. All re-infections were grouped under the "light infection" category. CONCLUSION: Mebendazole was found to be highly effective against A. lumbricoides, but had relatively low efficacy against hookworms and T. trichiura. These results bring into question the use of mebendazole in STH mass drug administration (MDA) programs in this region if albendazole, a drug with higher efficacy against hookworms, is available. Moreover, a significant number of treated children were re-infected with either or both of A. lumbricoides or hookworms 1 year after treatment emphasizing the need for better integrated intestinal helminthiasis control measures.

Clinical, parasitological and molecular profiles of Cutaneous Leishmaniasis and its associated factors among clinically suspected patients attending Borumeda Hospital, North-East Ethiopia.

BACKGROUND: Cutaneous leishmaniasis is one of the most neglected tropical diseases increasing in its public health importance. In Ethiopia over 28 million people are living at risk of infection. METHOD: Institution based cross-sectional study was conducted at Borumeda Hospital from February to May 2019. A total 205 leishmaniasis suspected patients were included by systematic random sampling technique. Socio demographic characteristics were collected using pre-tested questionnaires. Parasitological investigation was done from skin slit sample by using Geimsa staining method. Species identification was done by PCR-RFLP. Data were entered in to EpiData version 3.1 and analyzed using SPSS version 20 software. P-value of ≤ 0.05 was considered as statistically significant. RESULT: A total of 205 participants consisting 59% male and 41% female included in this study. The mean age (±SD) of the study participants was 31.9 (±14.29). The overall prevalence of cutaneous leishmaniasis was 22.4% (46/205). The prevalence in males (13.7%) was higher than in females (8.8%). It was more prevalent in the age group 16-45years old (15.6%). Clinically, 60% of patients' hade single lesion with 1.55 average number of lesions. About 30.7% of patients' had indurated plaque type of lesion. Most of the lesions were found on head and face (59%). House near to farmland, presence of hyrax in the village and presence of other cutaneous leishmaniasis cases in the neighborhood were independent predicator of cutaneous leishmaniasis prevalence. L.aethopica was found to be the etiologic agent of cutaneous leishmaniasis in the study participants. CONCLUSION: The prevalence of cutaneous leishmaniasis was 22.4%, this alerts the need of intervention. It is statistically associated with house near to farm land, presence of other cutaneous leishmaniasis cases in the neighborhood and presence of hyrax in village. Head and face were the most common sites of lesion.

Trends and seasonal patterns in intestinal parasites diagnosed in primary health facilities in Northwest Ethiopia.

INTRODUCTION: Intestinal parasites have an insidious impact on human health. In response to high parasite frequencies in Northwest Ethiopia, mass drug administration (MDA) is provided for school children using albendazole/mebendazole (since 2007) and praziquantel (since 2015). The study objective was to assess trends and seasonal patterns of intestinal parasite infections in a context of MDA. METHODOLOGY: This was a descriptive study collecting routine data from laboratory registers in two health centres in Denbia district, Amhara region. Stool test results (wet-mount direct microscopy) from patients attending these centres between 2013 and 2018 were included. Frequencies of different parasite species were evaluated within and across the years and stratified by age and gender. RESULTS: From a total of 8002 stool test results, the overall parasite frequency was 53.3%; this proportion remained constant. The most frequently diagnosed soil-transmitted helminths (STH) were Ascaris lumbricoides (16.9%) and hookworm (3.9%). STH frequency varied over the years, but was similar at the beginning (20.0%) and the end (22.0%) of the six-year period. STH infections were more frequent in winter (December-February; 20.4%) than in other seasons (16.0%). The most frequently diagnosed protozoa were Entamoeba histolytica/dispar (18.5%) and Giardia lamblia (12.2%). The frequency of Giardia steadily increased from 9.6% in 2013 to 15.3% in 2018. E. histolytica/dispar peaked in summer and G. lamblia in autumn. CONCLUSIONS: Trends in routine laboratories may be a proxy for a status quo in the community. These findings suggest that higher MDA coverages and/or interventions beyond MDA are needed to reduce intestinal parasite-related morbidity.

Does mass drug administration affect Schistosoma mansoni infection trends in West Dembia district, Northwest Ethiopia?

INTRODUCTION: Schistosomiasis is one of the Neglected Tropical Diseases in Ethiopia. Since 2015, yearly school-based mass drug administration (MDA) using praziquantel has become the major control strategy. This study aimed to assess trends of Schistosoma mansoni infection in a high-endemic area in Northwest Ethiopia. METHODOLOGY: Data were extracted from routine laboratory logbooks at two health centers in West Dembia district, Amhara region, for the period 2013-2018. Wet-mount direct microscopy was used to diagnose intestinal parasites. Chi-square test was used to compare proportions of S. mansoni-positive results before and after the start of MDA with praziquantel, across sex, age groups, and seasons. RESULTS: Data of 8002 stool tests was extracted. The proportion of S. mansoni progressively decreased from 9.6% in 2013 to 4.1% in 2018 in the overall patient population and from 20.3% in 2013 to 8.8% in 2018 in school-aged children. However, a declining trend of S. mansoni was observed before the launch of MDA and remained constant after the start of the MDA. The positivity rate was significantly higher in males and in the 5-14 years age group. S .mansoni infection in school aged children showed significant seasonal variation. CONCLUSIONS: The declined trend of S. mansoni positivity rate is encouraging and may be related to the existence of intervention packages. Although the timing of MDA was related with low positivity rate of S. mansoni infection, it has not resulted in the expected beneficial effect. Therefore, the district health office should work on both MDA and other interventions.

Genetic diversity and genotype multiplicity of Plasmodium falciparum infection in patients with uncomplicated malaria in Chewaka district, Ethiopia.

BACKGROUND: Genetic diversity in Plasmodium falciparum poses a major threat to malaria control and elimination interventions. Characterization of the genetic diversity of P. falciparum strains can be used to assess intensity of parasite transmission and identify potential deficiencies in malaria control programmes, which provides vital information to evaluating malaria elimination efforts. This study investigated the P. falciparum genetic diversity and genotype multiplicity of infection in parasite isolates from cases with uncomplicated P. falciparum malaria in Southwest Ethiopia. METHODS: A total of 80 P. falciparum microscopy and qPCR positive blood samples were collected from study participants aged 6 months to 60 years, who visited the health facilities during study evaluating the efficacy of artemether-lumefantrine from September-December, 2017. Polymorphic regions of the msp-1 and msp-2 were genotyped by nested polymerase chain reactions (nPCR) followed by gel electrophoresis for fragment analysis. RESULTS: Of 80 qPCR-positive samples analysed for polymorphisms on msp-1 and msp-2 genes, the efficiency of msp-1 and msp-2 gene amplification reactions with family-specific primers were 95% and 98.8%, respectively. Allelic variation of 90% (72/80) for msp-1 and 86.2% (69/80) for msp-2 were observed. K1 was the predominant msp-1 allelic family detected in 20.8% (15/72) of the samples followed by MAD20 and RO33. Within msp-2, allelic family FC27 showed a higher frequency (26.1%) compared to IC/3D7 (15.9%). Ten different alleles were observed in msp-1 with 6 alleles for K1, 3 alleles for MAD20 and 1 allele for RO33. In msp-2, 19 individual alleles were detected with 10 alleles for FC27 and 9 alleles for 3D7. Eighty percent (80%) of isolates had multiple genotypes and the overall mean multiplicity of infection was 3.2 (95% CI 2.87-3.46). The heterozygosity indices were 0.43 and 0.85 for msp-1 and msp-2, respectively. There was no significant association between multiplicity of infection and age or parasite density. CONCLUSIONS: The study revealed high levels of genetic diversity and mixed-strain infections of P. falciparum populations in Chewaka district, Ethiopia, suggesting that both endemicity level and malaria transmission remain high and that strengthened control efforts are needed in Ethiopia.

Anopheles gambiae s.l (Diptera: Culicidae) seasonal abundance, abdominal status and parity rates in Metema-Armachiho lowland, Northwest Ethiopia.

BACKGROUND: Malaria is a life threating vector borne disease caused by different Plasmodium parasites. Metema and Armachiho are two of the top five malaria endemic areas among the districts of Amhara region in Ethiopia. Transmission pattern is seasonal and migrant laborers who visit these areas for employment in mechanized agriculture are highly affected. The aim of this study was to investigate seasonal abundance, abdominal status and parity rate of An.gambiae s.l in Metema-Armachiho lowlands, Northwest Ethiopia. METHOD: A 1 year longitudinal entomological study was conducted in Metema-Armachiho lowlands from June 2016-May 2017. Mosquitoes were collected using CDC-light traps in indoor and outdoor sites for four consecutive days in each month. A total of eight standard battery operated CDC-light traps were used to collect mosquitoes. Female mosquitoes were classed as unfed, fed or gravid under a dissecting microscope. The ovaries of all unfed An.gambiae s.l mosquitoes were examined for evidence of parity. Data were entered and analyzed using SPSS-20 software. Chi-square test was applied to show significant difference between variables. P-value< 0.05 was taken as statistically significant. RESULTS: Of the total 1253 mosquitoes collected, 713 (552 female, 161 male) were culex and 540 (501 female, 39 male) were An.gambiae s.l. About 50.9% were collected in June-August 2016, 21.7% in September-November 2016, 12.0% in December 2016-February 2017 and 15.4% in March to May 2017. Of the total, 57.2 and 42.8% of the An.gambiae s.l were collected from indoor and outdoor sites respectively. Of the total females collected, 76.8% were unfed; of which 69.4% were parous. Significantly higher number of female An.gambiae s.l were collected in indoor and there was significant difference in abdominal status of An.gambiae s.l mosquitoes collected in different season (P < 0.05). CONCLUSIONS: Highest number of An.gambiae s.l was observed from June-August followed by September-November. The parity rate of An.gambiae s.l was high and there was significant difference in abdominal status of An.gambiae s.l collected in different season.

Validation of in-house liquid direct agglutination test antigen: the potential diagnostic test in visceral Leishimaniasis endemic areas of Northwest Ethiopia.

BACKGROUND: Visceral leishmaniasis in Ethiopia is a re-emerging threat to public health, with increased geographical distribution and number of cases. It is a fatal disease without early diagnosis and treatment; thus, the availability of affordable diagnostic tools is crucial. However, due to delays caused by import regulations, procurement and late delivery of imported test kits, accessibility remains a problem in the control program. Therefore, we aimed to produce and evaluate the performance of an in-house liquid (AQ) direct agglutination test (DAT) antigen. RESULT: The AQ-DAT was produced at the Armauer Hansen Research Institute, using Leishmania donovani strain (MHOM/ET/67/L82). Sera from 272 participants; 110 microscopically confirmed cases of VL, 76 apparently healthy and 86 patients who had infectious disease other than VL were tested with AQ-DAT, and standard kits: Freeze-dried DAT (FD-DAT) and rK39. Taking microscopy as a gold standard; the sensitivity and specificity of the AQ-DAT were 97.3 and 98.8%, respectively. It had high degrees of agreement (k > 0.8), with a significant (P < 0.05) correlation compared to microscopy, FD-DAT, and rK39. CONCLUSION: Although further standardization is required, the in-house AQ-DAT could improve diagnostic accessibility, minimize intermittent stock outs and strengthen the national VL control program.

Prevalence and Reinfection Rates of Schistosoma mansoni and Praziquantel Efficacy against the Parasite among Primary School Children in Sanja Town, Northwest Ethiopia.

BACKGROUND: Schistosomiasis is among the most widespread chronic infections in the world. The magnitude of the infection may show variations across different areas with respect to time. Praziquantel is a first line drug of choice for the treatment of schistosomiasis although its low cure rate has been reported in different parts of the world. Thus, an assessment of the magnitude of the diseases, the efficacy of currently available drugs, and reinfection rates is crucial. OBJECTIVE: Our principal objective is to determine the prevalence and reinfection rates of Schistosoma mansoni and to evaluate the efficacy of PZQ against Schistosoma mansoni. METHOD: A school-based cross-sectional study was conducted on Sanja Elementary Schools, Sanja town, northwest Ethiopia. Stool specimens were examined using Kato-Katz method. Schoolchildren who tested positive for intestinal schistosomiasis and fulfilled the inclusion criteria took part in the efficacy and reinfection study. Positive participants were treated with 40 mg/kg of Praziquantel. Cure and egg reduction rates were evaluated three weeks after treatment. The intensity of infection was determined following the WHO's guideline. Moreover, the reinfection rate of those who were cured was evaluated after a six-month posttreatment period. Data were analyzed using SPSS version 20. RESULTS: At baseline, 130 (35%) of the 372 schoolchildren were found infected with Schistosoma mansoni. Out of the 130 infected schoolchildren, 112 (86.2%) had moderate infection intensity. Among the S. mansoni positive schoolchildren, 80 were included as study participants for the evaluation of PZQ efficacy, based on the inclusion criteria established by WHO. The cure and egg reduction rates were found to be 90% (72/80) and 99.5%, respectively. Of the seventy-two schoolchildren considered for the determination of reinfection rate, after 6 months of posttreatment, 13.9% were found to be reinfected. CONCLUSION: The schoolchildren in the three primary schools of Sanja are at moderate risk of the infection caused by S. mansoni. Although the therapeutic potency of PZQ at 40 mg/kg was efficient against S. mansoni, a high rate of reinfection was reported in the study site, suggesting the need for integrated schistosomiasis control measures.

Molecular detection of Enteropathogens from diarrheic stool of HIV positive patients in Gondar, Ethiopia.

BACKGROUND: Infectious diarrhea is a common problem in the developing world, especially among people living with HIV/AIDS. Traditional diagnostic methods such as stool culture and microscopic examination are limited by resources and poor sensitivity. The use of molecular diagnostics for enteropathogen detection in this region of sub-Saharan Africa has not been fully explored. We sought to identify risk factors and characterize enteropathogens from diarrheic stools of HIV-positive patients in Gondar, Ethiopia using multiplex molecular panels targeting key infectious agents. METHODS: A cross-sectional study of 100 stool samples was performed. Samples were collected consecutively from HIV- positive patients presenting with diarrhea at University of Gondar Hospital clinic, a major center in NW Ethiopia. Genomic DNA was extracted from stool and processed using a multiplex molecular panel Allplex™ [Seegene, Canada]. Correlations between patient characteristics, symptoms, public health risk factors, and enteropathogen type (s) were studied. Eighty-six samples were successfully analyzed by molecular methods. RESULTS: The mean age was 35 with 43% male. Eighty percent lived in an urban area, 18% had access to well water only, and 81% practiced proper hand hygiene. The majority of patients (72%) were receiving HAART with a median CD4 cell count of 362/µL. Multiple pathogens were detected in 94% of specimens, with an average of 5 enteropathogens per sample. Common bacteria, viruses, and parasites detected were Shigella spp./enteroinvasive E. coli (80%), enterotoxigenic E. coli (73%), Norovirus (16%) and B. hominis (62%). CD4 cell count < 500/ µL was associated with the presence of viruses (p = 0.004) and the absence of STEC (p = 0.010). The use of HAART or CD4 levels was not associated with the number of enteropathogens detected. CONCLUSIONS: Diarrheic stool from HIV-positive outpatients in Gondar, Ethiopia had on average 5 enteropathogens present in their stool. Shigellaspp./enteroinvasive E. coli and enterotoxigenic E. coli are the major pathogens, not dissimilar to immunocompetent individuals in low income countries.

A 17-year trend analysis of malaria at Adi Arkay, north Gondar zone, Northwest Ethiopia.

BACKGROUND: Malaria is one of the leading causes of death worldwide. This study aimed to determine the trend of malaria among febrile patients seeking treatment over 17 year (1997-2013) at Adi Arkay, Northwest Ethiopia. METHODS: A 17-year malaria microscopy data were extracted retrospectively at Adi Arkay health centre. Time series and curve estimation analysis were used to evaluate trends in the data. Pearson's Chi square test was also used to describe associations of variables. RESULTS: Over 17 years, 20,483 blood films were requested for malaria diagnosis at the health centre. Out of this, 7428 (36.1%) were microscopically confirmed malaria cases. Plasmodium falciparum, Plasmodium vivax, and their mixed infection accounted for 68.85, 28.79, and 2.34% of all malaria cases, respectively. There was a remarkable reduction of overall malaria during the 17 years. Malaria was reported in all age groups of both sexes, but its positivity rate was significantly higher in males and in the 15-24 years than their counterparts. CONCLUSION: In relative terms, the overall positivity rate of malaria in the area over 17 years showed a significant reduction, but its magnitude as a public health problem is still alarming. Plasmodium falciparum played a significant role in the remarkable drop of overall malaria in the area, whereas vivax malaria remained unchanged. Therefore, control measures should continue to strengthen targeting both predominant malaria parasites in the area.