Plasmodium falciparum transmission in the highlands of Ethiopia is driven by closely related and clonal parasites.

Malaria cases are frequently recorded in the Ethiopian highlands even at altitudes above 2000 m. The epidemiology of malaria in the Ethiopian highlands, and, in particular, the role of importation by human migration from the highly endemic lowlands is not well understood. We sequenced 187 Plasmodium falciparum samples from two sites in the Ethiopian highlands, Gondar (n = 159) and Ziway (n = 28), using a multiplexed droplet digital PCR (ddPCR)-based amplicon sequencing method targeting 35 microhaplotypes and drug resistance loci. Here, we characterize the parasite population structure and genetic relatedness. We identify moderate parasite diversity (mean HE : 0.54) and low infection complexity (74.9% monoclonal). A significant percentage of infections share microhaplotypes, even across transmission seasons and sites, indicating persistent local transmission. We identify multiple clusters of clonal or near-clonal infections, highlighting high genetic relatedness. Only 6.3% of individuals diagnosed with P. falciparum reported recent travel. Yet, in clonal or near-clonal clusters, infections of travellers were frequently observed first in time, suggesting that parasites may have been imported and then transmitted locally. 31.1% of infections are pfhrp2-deleted and 84.4% pfhrp3-deleted, and 28.7% have pfhrp2/3 double deletions. Parasites with pfhrp2/3 deletions and wild-type parasites are genetically distinct. Mutations associated with resistance to sulphadoxine-pyrimethamine or suggested to reduce sensitivity to lumefantrine are observed at near-fixation. In conclusion, genomic data corroborate local transmission and the importance of intensified control in the Ethiopian highlands.

Dry-season transmission and determinants of Plasmodium infections in Jawi district, northwest Ethiopia.

BACKGROUND: Malaria remains a serious global public health problem, and continues to have a devastating impact on people's health worldwide. Continuous monitoring and evaluation of current malaria transmission status in different seasons is a mainstay for the success of ongoing intervention strategies for malaria. The purpose of this study was to assess the dry-season transmission and determinants of malaria in Jawi district, northwest Ethiopia. METHODS: A community-based cross-sectional study was conducted from January 13 to February 11, 2020; among selected Kebeles in the Jawi district. A multistage sampling technique was used in this study. Random and systematic sampling techniques were carried out to select Kebeles and each household, respectively. Light microscopy and CareStart™ Malaria HRP2/pLDH (Pf/Pv) Combo RDT were implemented to determine the prevalence of malaria. Moreover, associated risk factors in the prevalence of malaria were assessed by using a bivariate and multivariate logistic regression model. RESULTS: A total of 219 study participants were enrolled in this study. Of the total enrolled individuals, malaria cases were found among 36 individuals with a positivity rate of 16.4% (95% CI 11.4-21.5). Plasmodium falciparum was the predominant species with an estimated prevalence of 87.0% in the study areas. Interrupted utilization of ITN (AOR = 4.411, 95% CI 1.401-13.880), using over 3 years older ITNs (AOR = 9.622, 95% CI 1.881-49.214), travel history (AOR = 12.703, 95% CI 2.441-66.114), living in a house with holes on the wall (AOR = 3.811, 95% CI 1.010-14.384), and living in a house with an eave (AOR = 4.23, 95% CI 1.065-16.801) significantly increased the probability of malaria positivity rate. CONCLUSION: Malaria is still an important public health burden among individuals in the Jawi district. Interrupted utilization of ITNs, using over 3 years older ITNs, living in a house with holes on the wall, living in a house with an eave, and travel history were identified as the risk factors of malaria. Therefore, the District health office and Health extension workers should promote daily utilization of good ITNs and improve housing conditions to reduce malaria prevalence.

An update on the distribution, bionomics, and insecticide susceptibility of Anopheles stephensi in Ethiopia, 2018-2020.

BACKGROUND: Anopheles stephensi, an invasive malaria vector, was first detected in Africa nearly 10 years ago. After the initial finding in Djibouti, it has subsequently been found in Ethiopia, Sudan and Somalia. To better inform policies and vector control decisions, it is important to understand the distribution, bionomics, insecticide susceptibility, and transmission potential of An. stephensi. These aspects were studied as part of routine entomological monitoring in Ethiopia between 2018 and 2020. METHODS: Adult mosquitoes were collected using human landing collections, pyrethrum spray catches, CDC light traps, animal-baited tent traps, resting boxes, and manual aspiration from animal shelters. Larvae were collected using hand-held dippers. The source of blood in blood-fed mosquitoes and the presence of sporozoites was assessed through enzyme-linked immunosorbent assays (ELISA). Insecticide susceptibility was assessed for pyrethroids, organophosphates and carbamates. RESULTS: Adult An. stephensi were collected with aspiration, black resting boxes, and animal-baited traps collecting the highest numbers of mosquitoes. Although sampling efforts were geographically widespread, An. stephensi larvae were collected in urban and rural sites in eastern Ethiopia, but An. stephensi larvae were not found in western Ethiopian sites. Blood-meal analysis revealed a high proportion of blood meals that were taken from goats, and only a small proportion from humans. Plasmodium vivax was detected in wild-collected An. stephensi. High levels of insecticide resistance were detected to pyrethroids, carbamates and organophosphates. Pre-exposure to piperonyl butoxide increased susceptibility to pyrethroids. Larvae were found to be susceptible to temephos. CONCLUSIONS: Understanding the bionomics, insecticide susceptibility and distribution of An. stephensi will improve the quality of a national response in Ethiopia and provide additional information on populations of this invasive species in Africa. Further work is needed to understand the role that An. stephensi will have in Plasmodium transmission and malaria case incidence. While additional data are being collected, national programmes can use the available data to formulate and operationalize national strategies against the threat of An. stephensi.

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.

Impact of high malaria incidence in seasonal migrant and permanent adult male laborers in mechanized agricultural farms in Metema - Humera lowlands on malaria elimination program in Ethiopia.

BACKGROUND: Seasonal migrant and permanent laborers who are working in big mechanized agricultural farms in Metema - Humera lowlands are not included in Ethiopia Malaria Elimination Program. The aim of this research was to show the high confirmed and treated malaria cases in these laborers. METHODS: A retrospective analysis of the confirmed and treated malaria cases in all the districts of West, Central and North Gondar Zones, using Weekly Public Health Emergency Management (PHEM) reports, was conducted to show a complete picture of the malaria incidences in the areas. RESULT: A total of 3,485,646 confirmed malaria cases were treated in Amhara region during 2013 to 2017. Of the total malaria cases in the Amhara region during these period, 1, 286, 848 cases or 37.2% were originated from West, Central and North Gondar Zones. But these 3 Zones contribute only 17% of Amhara region population. Of all the confirmed malaria cases reported in the 3 Zones, 41.7% (536,749/1286, 848) was reported from the three lowland districts (Metema, West Armachiho and Quara) of the West Gondar Zone during the same study period. But, the West Gondar Zone has only around 10% of the population in these three zones. The highest annual parasite incidence for malaria was found in West Armachiho district. Majority of above 14 years malaria cases in West Gondar zone were found from laborers. CONCLUSION: Migrant and permanent laborers working in mechanized agricultural fields in Metema - Humera lowlands are highly exposed to malaria and immediate interventions are required.

Malaria-visceral leishmaniasis co-infection and associated factors among migrant laborers in West Armachiho district, North West Ethiopia: community based cross-sectional study.

BACKGROUND: Malaria and leishmaniasis are the two largest parasitic killers in the world. Due togeographical overlap of these diseases, malaria-visceral leishmaniasis co-infections occur in large populations and exist in different areas even if they have been poorly investigated. The aim of this study was to determine malaria-visceral leishmaniasis co-infection and their associated factors among migrant laborers. METHODS: Community based cross-sectional study was conducted from October-December 2016 on migrant laborers who are residents of rural agricultural camp in West Armachiho district and involved in sesame and sorghum harvesting. Standardized questionnaire was used to collect socio-demographic data and risk factors. Capillary blood was collected for giemsa stained blood film examination to detect and identify Plasmodium parasites. Recombinant kinensin (rk39) antigen test was performed to detect anti-leishmania donovani antibody. Data was coded, entered, checked for completeness and analyzed using SPSS version-20 statistical software. Chi-square test was applied to show a significant association between variables. P-value < 0.05 was considered as statistically significant. RESULTS: A total of 178 migrant laborers were included in this study. Of these, 74.2% belong to the age group 15-29; 61.2% come from lowland areas and 51.6% visit the area more than four times. Seroprevalence of visceral leishmaniasis was 9.6% (17/178); and 22.4% (40/178) of tested migrant laborers were found malaria infected. The overall prevalence of malaria-visceral leishmaniasis co-infection was 2.8%. Of the total migrant laborer, 47.8% used bed nets, of them 1.2% were malaria-visceral leishmaniasis co-infected; 72.5% used outdoor sites as usual sleeping site, among them 3.1% were malaria-visceral leishmaniasis co-infected; 60.1% were migrants, of which 2.8% were malaria-visceral leishmaniasis co-infected. All variables were not significantly associated with malaria-visceral leishmaniasis co-infection (P > 0.05). CONCLUSIONS: Prevalence of malaria-visceral leishmaniasis co-infection was low and it is not significantly associated with residence, number of visits, bed net utilization and outdoor sleeping habit even if both diseases are prevalent in the study area.

Performance of loop-mediated isothermal amplification (LAMP) for the diagnosis of malaria among malaria suspected pregnant women in Northwest Ethiopia.

BACKGROUND: Malaria is a major public health problem and an important cause of maternal and infant morbidity in sub-Saharan Africa, including Ethiopia. Early and accurate diagnosis of malaria with effective treatment is the best strategy for prevention and control of complications during pregnancy and infant morbidity and mortality. However, laboratory diagnosis has relied on the identification of malaria parasites and parasite antigens in peripheral blood using Giemsa-stained microscopy or rapid diagnostic tests (RDTs) which lack analytical and clinical sensitivity. The aim of this study was to evaluate the performance of loop-mediated isothermal amplification (LAMP) for the diagnosis of malaria among malaria suspected pregnant women in Northwest Ethiopia. METHODS: A cross sectional study was conducted from January to April 2016. Pregnant women (n = 87) suspected of having malaria at six health centres were enrolled. A venous blood sample was collected from each study subject, and analysed for Plasmodium parasites by microscopy, RDT, and LAMP. Diagnostic accuracy outcome measures (sensitivity, specificity, predictive values, and Kappa scores) of microscopy, RDT and LAMP were compared to nested polymerase chain reaction (nPCR) as the gold standard. Specimen processing and reporting times were documented. RESULTS: Using nPCR as the gold standard technique, the sensitivity of microscopy and RDT was 90 and 70%, and the specificity was 98.7 and 97.4%, respectively. LAMP assay was 100% sensitive and 93.5% specific compared to nPCR. CONCLUSIONS: This study showed higher sensitivity of LAMP compared to microscopy and RDT for the detection of malaria in pregnancy. Increased sensitivity and ease of use with LAMP in point-of-care testing for malaria in pregnancy was noted. LAMP warrants further evaluation in intermittent screening and treatment programmes in pregnancy.

Sero-prevalence of Leishmania donovani infection in labour migrants and entomological risk factors in extra-domestic habitats of Kafta-Humera lowlands - kala-azar endemic areas in the northwest Ethiopia.

BACKGROUND: Visceral leishmaniasis (VL) or kala-azar cases in seasonal labour migrants from highland areas are addressed to travel history to the Metema-Humera lowlands, northwestern Ethiopia. Factors that affect the incidence of VL in extra-domestic habitats were not evaluated. The aim of this study was to evaluate sero-prevalence of Leishmania donovani infection in randomly selected labour migrant workers and entomological risk factors which might affect the incidence of kala-azar. METHODS: Sero-prevalence of L. donovani infection in labour migrants was obtained from Direct Agglutination Test (DAT) using blood samples. Logistic regression analysis was used to correlate the possible risk factors with L. donovani infection. The season for L. donovani infection or Phlebotomus orientalis bite was estimated from the study of population dynamic of P. orientalis in areas where the blood was sampled. RESULT: A total of 7, 443 P. orientalis (1,748 female and 5,695 male) were collected from agricultural fields and thickets of Acacia seyal using 461 CDC light traps. The highest mean number of P. orientalis/trap in the thickets of A. seyal and agricultural fields were 46.9 and 43.9 in March and April respectively. The mean P. orientalis/trap for November - May dry season in agricultural fields (11.39) and thickets of A. seyal (25.30) were higher compared to 0.66 in fields and 3.92 in thickets during June - August weeding season. Of the total 359 labour migrants screened using DAT, 45 (12.5%) were DAT-positive (≥1:800) for L. donovani infections. Very high titers (1:12800) were found in 3 (0.8%) individuals who had the risk of kala-azar development. Statistically significant p-values and odd ratio (OR) for staying in the areas both in the weeding and harvesting seasons (p = 0.035; OR = 2.83) and sleeping in the agricultural fields (p = 0.01; OR = 15.096) were positively correlated with L. donovani infection. Night harvest (p = 0.028; OR = 0.133) and knowledge about sign or symptoms (p = 0.042; OR = 0.383) were negatively associated with this infection. CONCLUSIONS: Sleeping in open agricultural fields was related with L. donovani infections in labour migrants during June-August weeding season.

Assessment of the selected hematological profiles among malaria and Schistosoma mansoni co-infected patients, Northwest Ethiopia.

Malaria and schistosomiasis are infectious diseases that cause hematological profiles abnormalities. Malaria and Schistosoma mansoni co-infection causes exacerbation of health consequences and co-morbidities. The aim of this study was to assess the selected hematological profiles among malaria and S. mansoni co-infected patients at Dembiya Selected Health Institutions. An institutional-based comparative cross-sectional study was conducted from March 30 to August 10, 2022. A total of 140 individuals were enrolled in the study using a convenient sampling technique. Wet mount and Kato Katz technique were conducted to detect S. mansoni in Stool sample. Blood films were prepared for the detection of plasmodium. The data was coded and entered into EpiData version 3.1 before being analyzed with SPSS version 25. A P-value of less than 0.05 was considered statistically significant. Mean of WBC, neutrophil, lymphocyte, eosinophil, RBC, hemoglobin, and hematocrit [4.IU/L,2.2 IU/L, 1.4 IU/L, 0.1 IU/L, 3.13 IU/L, 9.5 g/dL, and 28.7%, resepectively] in co-infected were significantly lower than [7.5 IU/L, 4.6 IU/L, 2.1 IU/L, 0.38 IU/L, 4.8 IU/L, 14.6 g/dL, and 43.7%, resepectively] in the healthy control participants. Mean of RBC, hemoglobin, and hematocrit [3.13 IU/L, 9.5 g/dL, 28.7%, resepectively] in co-infected were significantly lower compared to [3.8 IU/L, 11.5 g/dL, 33.9%, resepectively] in the malaria monoinfected participants and [3.7 IU/L,11.5 g/dL, 33.6%, resepectively] in the S. mansoni monoinfected participants. The result of hematological profiles in healthy participants had no significant difference compared to light,moderate and heavy S. mansoni infection intensity in coinfection. The number of S. mansoni eggs per gram of stool had been negatively correlated with hematological profiles of co-infected participants except lymphocyte and monocyte which correlated positively. Hematological profiles status in coinfection were significantly altered compared to malaria monoinfection, S. mansoni monoinfection, and healthy participants.Therefore, hematological tests should be used to monitor and manage coinfection related complications, and to reduce coinfection associated morbidity and mortality.

Effect of Malaria and Schistosoma mansoni Coinfection on Selected Biochemical Profiles among Patients Attending Selected Health Institutions at Dembiya, Northwest Ethiopia.

Background: Malaria and schistosomiasis are infectious diseases that cause biochemical abnormalities. Malaria and Schistosoma mansoni coinfection causes exacerbations of health consequences and comorbidities. The study area is found in Ethiopia, where coinfection of malaria and S. mansoni is common. However, there is limited data on the biochemical profiles of patients coinfected with malaria and S. mansoni schistosomiasis in the study area. Hence, this study aimed to assess the effect of malaria and S. mansoni schistosomiasis coinfection on selected biochemical profiles. Methods: An institutional-based comparative cross-sectional study was conducted from March 30 to August 10, 2022. Using a convenient sampling technique, 70 participants (35 cases and 35 controls) were enrolled in the study. Schistosoma mansoni was detected in stool samples using the wet mount and the Kato Katz method. To detect Plasmodium, both thick and thin blood films were prepared and stained with Giemsa. The blood sample was processed for the analysis of biochemical profiles. All data were analyzed using SPSS version 25. A p value of less than 0.05 was considered statistically significant. Results: The mean values of alanine aminotransferase and aspartate aminotransferase (37.1 U/L and 41.9 U/L, respectively) in coinfected participants were significantly higher than in the healthy control participants (17.4 U/L and 22.0 U/L, respectively) (p < 0.05). Also, the median values of creatinine, total bilirubin, and direct bilirubin (1.51 mg/dL, 2.35 mg/dL, and 0.91 mg/dL, respectively) in coinfected participants were significantly higher than in the healthy control participants (0.85 mg/dL, 0.42 mg/dL, and 0.12 mg/dL, respectively) (p < 0.05). However, median values of total protein (4.82 g/dL) and mean values of glucose (66.6 mg/dL) in coinfected participants were significantly lower than in the healthy control participants (total protein (7.64 g/dL) and glucose (91.9 mg/dL)) (p < 0.05). The results of biochemical profiles in healthy participants were significantly different from those with light, moderate, and heavy S. mansoni infection intensity in malaria and S. mansoni coinfection (p < 0.05). Schistosoma mansoni infection intensity had a positive correlation with biochemical profiles except for total protein and glucose, which correlated negatively in coinfected participants (p > 0.05). Conclusion: Biochemical profiles in coinfection were significantly changed as compared to healthy individuals. As a result, biochemical profile tests should be utilized to monitor and manage coinfection-related problems, as well as to reduce coinfection-related morbidity and death.

Status of selected biochemical and coagulation profiles and platelet count in malaria and malaria-Schistosoma mansoni co-infection among patients attending at Dembiya selected Health Institutions, Northwest Ethiopia.

Malaria and schistosomiasis are infectious diseases that cause coagulation disorders, biochemical abnormalities, and thrombocytopenia. Malaria and Schistosoma mansoni co-infection cause exacerbations of health consequences and co-morbidities.This study aimed to compare the effect of malaria and Schistosoma mansoni co-infection and malaria infection on selected biochemical and coagulation profiles, and platelet count. An institutional-based comparative cross-sectional study was conducted from March 30 to August 10, 2022. A total of 70 individuals were enrolled in the study using a convenient sampling technique. Wet mount and Kato Katz techniques were conducted to detect Schistosoma mansoni in a stool sample. Blood films were prepared for the detection of plasmodium. The data was coded and entered into EpiData version 3.1 before being analyzed with SPSS version 25. An independent t test was used during data analysis. A P-value of less than 0.05 was considered statistically significant. The mean [SD] of alanine aminotransferase, aspartate aminotransferase, creatinine, total bilirubin, and direct bilirubin in the co-infected was higher than in malaria infected participants. However, the mean of total protein and glucose in co-infected was lower than in the malaria infected participants. The mean of prothrombin time, international normalization ratio, and activated partial thromboplastin time in co-infected was significantly higher, while the platelet count was lower compared to malaria infected participants. Biochemical and coagulation profiles, and platelet count status in co-infection were changed compared to malaria infected participants. Therefore, biochemical and coagulation profiles and platelet count tests should be used to monitor and manage co-infection related complications and to reduce co-infection associated morbidity and mortality.

A digital microscope for the diagnosis of Plasmodium falciparum and Plasmodium vivax, including P. falciparum with hrp2/hrp3 deletion.

Sensitive and accurate malaria diagnosis is required for case management to accelerate control efforts. Diagnosis is particularly challenging where multiple Plasmodium species are endemic, and where P. falciparum hrp2/3 deletions are frequent. The Noul miLab is a fully automated portable digital microscope that prepares a blood film from a droplet of blood, followed by staining and detection of parasites by an algorithm. Infected red blood cells are displayed on the screen of the instrument. Time-to-result is approximately 20 minutes, with less than two minutes hands-on time. We evaluated the miLab among 659 suspected malaria patients in Gondar, Ethiopia, where P. falciparum and P. vivax are endemic, and the frequency of hrp2/3 deletions is high, and 991 patients in Ghana, where P. falciparum transmission is intense. Across both countries combined, the sensitivity of the miLab for P. falciparum was 94.3% at densities >200 parasites/µL by qPCR, and 83% at densities >20 parasites/µL. The miLab was more sensitive than local microscopy, and comparable to RDT. In Ethiopia, the miLab diagnosed 51/52 (98.1%) of P. falciparum infections with hrp2 deletion at densities >20 parasites/µL. Specificity of the miLab was 94.0%. For P. vivax diagnosis in Ethiopia, the sensitivity of the miLab was 97.0% at densities >200 parasites/µL (RDT: 76.8%, microscopy: 67.0%), 93.9% at densities >20 parasites/µL, and specificity was 97.6%. In Ethiopia, where P. falciparum and P. vivax were frequent, the miLab assigned the wrong species to 15/195 mono-infections at densities >20 parasites/µL by qPCR, and identified only 5/18 mixed-species infections correctly. In conclusion, the miLab was more sensitive than microscopy and thus is a valuable addition to the toolkit for malaria diagnosis, particularly for areas with high frequencies of hrp2/3 deletions.

Antibacterial effects of Apis mellifera and stingless bees honeys on susceptible and resistant strains of Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae in Gondar, Northwest Ethiopia.

BACKGROUND: Honey is a natural substance produced by honeybees and has nutritional and therapeutic uses. In Ethiopia, honeys are used traditionally to treat wounds, respiratory infections and diarrhoea. Recent increase of drug resistant bacteria against the existing antibiotics forced investigators to search for alternative natural remedies and evaluate their potential use on scientific bases. Thus, the aim of this study was to evaluate the antibacterial effects of different types of honeys in Ethiopia which are used traditionally to treat different types of respiratory and gastrointestinal infections. METHODS: Mueller Hinton agar (70191) diffusion and nutrient broth culture medium assays were performed to determine susceptibility of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and resistant clinical isolates (Methicillin resistant Staphylococcus aureus(MRSA), Escherichia coli(R) and Klebsiella pneumoniae (R), using honeys of Apis mellifera and stingless bees in northern and north western Ethiopia. RESULTS: Honey of the stingless bees produced the highest mean inhibition (22.27 ± 3.79 mm) compared to white honey (21.0 ± 2.7 mm) and yellow honey (18.0 ± 2.3 mm) at 50% (v/v) concentration on all the standard and resistant strains. Stingless bees honey was found to have Minimum Inhibitory Concentration (MIC) of 6.25% (6.25 mg/ml) for 80% of the test organisms compared to 40% for white and yellow Apis mellifera honeys. All the honeys were found to have minimum bactericidal concentration (MBC) of 12.5% (12.5 mg/ml) against all the test organisms. Staphylococcus aureus (ATCC 25923) was susceptible to amoxicillin, methicillin, kanamycine, tetracycline, and vancomycine standard antibiotic discs used for susceptibility tests. Similarly, Escherichia coli (ATCC 25922) was found susceptible for kanamycine, tetracycline and vancomycine. Escherichia coli (ATCC 25922) has not been tested for amoxicillin ampicillin and methicillin. The susceptibility tests performed against Staphylococcus aureus (MRSA), Escherichia coli (R) and Klebsiella pneumoniae (R) using three of methicillin, erythromycin, ampicillin, Penicillin and amoxicillin discs were resistant. But, these drug resistant strains were susceptible to antibacterial agents found in the honeys and inhibited from 16 mm to 20.33 mm. CONCLUSIONS: Honeys in Ethiopia can be used as therapeutic agents for drug resistant bacteria after pharmaceutical standardization and clinical trials.

Highland Malaria Transmission Dynamics in Space and Time Before Pre-elimination Era, Northwest Ethiopia.

BACKGROUND: Currently, the district-level malaria transmission stratification has indicated the Northern, Northwestern, Southern, and rift valley lowland and surrounding highland districts are almost entirely classified as high or moderate malaria transmission zones. Conducting malaria surveillance to track, test, and treat all malaria cases cannot be implemented in Ethiopia in the current situation. OBJECTIVE: To show malaria transmission dynamics in different health facilities located from 1800 to 2772 m altitudes during 2018-2021 in Northwest Ethiopia. METHODS: A total of 3.5 years (2018-2021) retrospective confirmed and treated malaria cases in 43 kebeles health posts and clinics in Gondar Zuria district were used for analysis. RESULT: The total malaria count was 5893 for 2019 compared to 31, 550 for 2020 and 33, 248 for 2021. Mean monthly malaria incidence/1000 people in 2019 was 2.39 ± 5.4 and increased to 10.64 ± 16.99 in 2020 and 11.19 ± 16.59 in 2021. Annual malaria incidence increased from 24 cases/1000 people in 2019 to 139.08 cases/1000 people in 2021 and is alarming danger in malaria elimination program in the district or the country as a whole. Poisson and Negative binomial regressions models indicated 5.78- and 5.26-fold malaria cases increase, respectively, in 2021 compared to 2019. The sudden increase in malaria incidences (counts) in 2020 and 2021 coincided with the interruption of residual insecticide application in Gondar Zuria district during the transition period towards the malaria pre-elimination stage implicating the role of malaria control tools in suppressing transmission. Study on climate variability also indicated that the rainfall variability in different months might have also favored high malaria transmission in 2020 and 2021 compared to 2019. Thus, in addition to re-starting the use of malaria control tools, giving attention to climate anomalies (variability) that favors malaria transmission, for prompt interventional actions, is required. The malaria elimination program in Ethiopia might have not reached a pre-elimination stage as malaria cases per 1000 people have not decreased below five in the majority of Ethiopian districts. Tracing, confirming, and treating individual cases to stop further transmission is, almost, impossible. In a situation like this, the Ethiopian malaria elimination program should work intensively towards understanding malaria epidemiology at the district level to re-design a localized malaria control strategy. The renewed malaria control program should also consider altitudes above 2000 m.

Description of malaria epidemics and normal transmissions using rainfall variability in Gondar Zuria highland District, Ethiopia.

BACKGROUND: Rainfall is one of the climate variables most studied as it affects malaria occurrence directly. OBJECTIVE: This study aimed to describe how monthly rainfall variability affects malaria incidence in different years. METHODS: A total of 7 years (2013/14-2019/20) retrospective confirmed and treated malaria cases in Gondar Zuria district were used for analysis in addition to five (2013/14-2017/18) years retrospective data from Dembia district. RESULTS: The annual rainfalls in the study years showed no statistically significant difference (p = 0. 78). But, variations in rainfalls of the different months (p = 0.000) of the different years were the source of variations for malaria count (incidences) in the different years. Malaria was transmitted throughout the year with the highest peak in November (mean count = 1468.7 ± 697.8) and followed by May (mean count = 1253.4 ± 1391.8), after main Kiremt/Summer and minor Bulg/Spring rains respectively. The lowest transmission was occurred in February (338 ± 240.3) when the rivers were the only source of mosquito vectors. Year 2013/14 (RF = 2351.12 mm) and 2019/20 (RF = 2278.80 mm) with no statistically significant difference (p = 0.977) in annual rainfalls produced 10, 702 (49.2%) and 961 (20%) malaria counts for the Bulg (spring) season respectively due to 581.92 mm (24.8%) higher total Bulg/Spring rain in 2013/14 compared to 124.1 mm (5.45%) in 2019/20. Generally, above normal rainfalls in Bulg/Spring season increased malaria transmission by providing more aquatic habitats supporting the growth of the immature stages. But heavy rains in Summer/Kiremt produced low malaria counts due to the high intensity of the rainfalls which could kill the larvae and pupae. Spearman's correlation analysis indicated that the mean rainfalls of current month (RF) (0 lagged month) (P = 0.025), previous month (RF1) (1 month lagged) (p = 0.000), before previous months (RF2) (2 months lagged) (p = 0.001) and mean RF + RF1 + RF2 (P = 0.001) were positive significantly correlated with mean monthly malaria counts compared to negative significant correlations for temperature variables. Temperature variables negative correlations were interpreted as confounding effects because decreased malaria counts in dry months were due to a decrease in rainfalls. Conclusion: rainfall distribution in different months of a year affects malaria occurrences.

Malaria Parasitemia in Febrile Patients Mono- and Coinfected with Soil-Transmitted Helminthiasis Attending Sanja Hospital, Northwest Ethiopia.

BACKGROUND: Malaria is a life-threatening disease associated with high morbidity and mortality. Helminths are among the most widespread infectious agents prevalent in tropical and subtropical regions of the developing world. Malaria and soil-transmitted helminthiasis (STHs) are coendemic and major public health problems in Ethiopia. The effects of helminth coinfection on malaria parasitemia remained poorly understood. Therefore, the objective of this study was to assess malaria parasitemia among malaria-monoinfected and malaria-soil-transmitted helminthiasis-coinfected febrile patients attending Sanja Hospital, Northwest Ethiopia. METHODS: A cross-sectional study with parallel groups was conducted to assess malaria parasitemia among malaria-monoinfected and malaria-soil-transmitted helminthiasis-coinfected febrile patients in Sanja Hospital from January to March 2019. Double population proportion formula was used for sample size calculation, and convenient sampling technique was used to select 134 study participants. Data were entered and analyzed by using the Statistical Package for Social Sciences (SPSS) version 20. Descriptive statistics, independent t-test, and one-way analysis of variance (ANOVA) were performed. A P value of <0.05 was considered as statistically significant. RESULTS: From 134 malaria-positive study participants, 67 were malaria-monoinfected and 67 were malaria-STHs-coinfected patients. Out of 67 malaria STHs-coinfected patients, 54 (80.6%) were infected with hookworm followed by Ascaris lumbricoides 11 (16.4%) and Strongyloides stercoralis 2 (3%). The mean Plasmodium parasite density was significantly higher in malaria-STHs-coinfected patients than in patients infected with only Plasmodium parasite density was significantly higher in malaria-STHs-coinfected patients than in patients infected with only P value of <0.05 was considered as statistically significant. Plasmodium parasite density was significantly higher in malaria-STHs-coinfected patients than in patients infected with only F = 6.953, P value of <0.05 was considered as statistically significant. CONCLUSIONS: Infections with STHs, especially hookworm, were positively associated with Plasmodium parasite density. The current study finding also revealed that increased worm burden of hookworm as expressed by egg intensity had significantly increased Plasmodium parasite density.Plasmodium parasite density was significantly higher in malaria-STHs-coinfected patients than in patients infected with only Plasmodium parasite density was significantly higher in malaria-STHs-coinfected patients than in patients infected with only.

Anopheles cinereus implicated as a vector of malaria transmission in the highlands of north-west Ethiopia.

BACKGROUND: Transmission of malaria in the highlands of Ethiopia is poorly understood and usually attributed to importation by mobile populations or local transmission by Anopheles arabiensis. To characterize and identify Anopheles species present in a highland area of northern Ethiopia, adult and larval collections were performed in Gondar town and the neighboring Senbet Debir village (Dembia district, > 2000 meters above sea level, masl), in addition to Bahir Dar town (capital of Amhara region) and Kumer Aftit village (Metema district, < 2000 masl). METHODS: CDC-light traps were used to collect adult mosquitoes and larval collections were performed from rain pools for rearing into adults for species identification. Collections were made September-March 2016-2018. Adult mosquitoes were identified morphologically and a subset of randomly chosen specimens were identified to species by sequencing the ribosomal DNA internal transcribed spacer region 2 (ITS2) and mitochondrial DNA cytochrome c oxidase subunit 1 (cox1). RESULTS: The primary species of Anopheles identified at elevations higher than 2000 masl was An. cinereus, which was confirmed molecularly by ITS2 and cox1 sequencing. Interestingly, two unknown species were also sequenced, in addition to two specimens of An. pretoriensis. The species collected at sites with elevations less than 2000 masl (Bahir Dar town and Kumer Aftit village) was An. arabiensis. Three Plasmodium falciparum-positive specimens were identified molecularly as An. cinereus. CONCLUSIONS: The presence of Plasmodium-positive An. cinereus in areas greater than 2000 masl incriminates this species as a potential vector contributing to non-peak malaria transmission in Ethiopian highland areas.

Seroprevalence of Asymptomatic Leishmania donovani among Laborers and Associated Risk Factors in Agricultural Camps of West Armachiho District, Northwest Ethiopia: A Cross-Sectional Study.

BACKGROUND: Visceral leishmaniasis (VL, also called kala-azar) is a public health problem in Ethiopia, especially in sesame and sorghum growing areas. Compared to other populations, labor migrants are the most exposed. Knowing the seroprevalence of Leishmania donovani and associated risk factors is essential to design appropriate control measures. The main aim of this study was to assess the seroprevalence of asymptomatic L. donovani among laborers and associated risk factors in agricultural camps of West Armachiho district, Northwest Ethiopia. Therefore, this study was conducted to know the seroprevalence and associated risk factors of L. donovani infection. METHOD: A cross-sectional study was conducted among 185 laborers from October to December 2017. A simple random sampling technique was used to select study participants from selected agricultural camps. After obtaining written informed consent, data were collected using a structured pretested Amharic version questionnaire using the interview technique. A single finger prick blood sample was collected from the study participants and the blood samples were subjected to the serological diagnostic method using the rk39 kit. The multivariable logistic regression model was used to identify risk factors associated with L. donovani infection. RESULT: Among 185 participants examined using rk39, 14 (7.6%) were seroreactive for L. donovani. Leishmania donovani infection had a statistically significant association with sleeping under Balanites trees (AOR: 4.36, 95%CI: 1.186-16.06), presence of domestic animals near sleeping place (AOR: 4.68, 95% CI: 1.25-17.56), and lack of knowledge about VL transmission (AOR: 3.79, 95% CI: 1.07-13.47). CONCLUSION: Seroprevalence of asymptomatic L. donovani among laborers in agricultural camps of West Armachiho was low. Prevention measures and health education about risk factors that expose to L. donovani infection for the laborers are essential to prevent the spread of the disease.