Ethiopian Plasmodium vivax hypnozoites formation dynamics and their susceptibility to reference antimalarial drugs.

One of the key obstacles to malaria elimination is largely attributed to Plasmodium vivax's ability to form resilient hypnozoites in the host liver that cause relapsing infections. As a result, interruption of P. vivax transmission is difficult. P. vivax transmission occurs in Duffy-positive individuals and have been mainly thought to be absent in Africa. However, increasing studies using molecular tools detected P. vivax among Duffy-negative individuals in various African countries. Studies on the African P. vivax has been severely limited because most of malaria control program focus mainly on falciparum malaria. In addition, there is a scarcity of laboratory infrastructures to overcome the biological obstacles posed by P. vivax. Herein, we established field transmission of Ethiopian P. vivax for routine sporozoite supply followed by liver stage infection in Mali. Furthermore, we evaluated local P. vivax hypnozoites and schizonts susceptibilities to reference antimalarial drugs. The study enabled the assessment of local African P. vivax hypnozoite production dynamics. Our data displayed the ability of the African P. vivax to produce hypnozoite forms ex-vivo at different rates per field isolate. We report that while tafenoquine (1µM) potently inhibited both hypnozoites and schizont forms; atovaquone (0.25µM) and the phosphatidylinositol-4-OH kinase (PI4K)-specific inhibitor KDU691 (0.5µM) showed no activity against hypnozoites forms. Unlike hypnozoites forms, P. vivax schizont stages were fully susceptible to both atovaquone (0.25µM) and the (PI4K)-specific inhibitor KDU691 (0.5µM). Together, the data revealed the importance of the local platform for further biological investigation and implementation of drug discovery program on the African P. vivax clinical isolates.

Infectivity of Symptomatic Patients and Their Contribution for Infectiousness of Mosquitoes following a Membrane Feeding Assay in Ethiopia.

The membrane feeding assay is widely used to evaluate the efficacy of transmission-blocking interventions (TBIs) and identify the reservoir of malaria. This study aimed to determine the infectivity of blood meals from symptomatic Plasmodium-infected patients to an Anopheles arabiensis colony in Ethiopia. A membrane feeding assay was conducted on a total of 63 Plasmodium falciparum- and/or Plasmodium vivax-infected clinical patients in East Shoa Zone, Ethiopia. Detection of P. falciparum and P. vivax in blood samples was done using microscopy. Mosquito infection rates were determined by dissection of mosquitoes' midguts, while mosquito infectiousness was observed by dissection of their salivary glands. The proportion of infectious symptomatic patients was 68.3% (43/63). Using the chi-square or Fisher's exact test, the oocyst infection levels were higher among patients infected with P. vivax, females, and rural residents. Nearly 57% (56.7%, 17/30) of assays produced sporozoites in the salivary glands of mosquitoes. Both oocyst and sporozoite infection rates had positive correlations with parasitemia and gametocytemia. High infectiousness of symptomatic patients was observed, with a greater proportion of infectious mosquitoes per assay. Demonstrating oocyst infection in the mosquitoes might confirm estimates of the infectiousness of mosquitoes, although some of the oocyst-infected mosquitoes failed to produce sporozoites. IMPORTANCE Malaria remains one of the most devastating infectious diseases globally, and transmission-blocking activities are needed. Plasmodium transmission from human to mosquitoes is poorly studied, particularly in endemic countries, and the membrane feeding assay allows it to be determined. In this study, we demonstrated human infectious reservoirs of malaria. Moreover, the effect of Plasmodium-infected patients on the infectiousness of mosquitoes was also observed. These findings are therefore important for designing future evaluation of transmission-blocking interventions that will support the malaria elimination program.

Predictors of unsuppressed viral load among adults on follow up of antiretroviral therapy at selected public and private health facilities of Adama town: unmached case-control study.

BACKGROUND: Despite the scale up of antiretroviral therapy (ART), unsuppressed viral load among population taking ART in private and public health facilities is still a public health concern increasing the risk of treatment failure. Studies comprehensively assessing significant predictors of non-suppressed viral load among patients on follow up of AR in public and private health facilities are limited. The objective of the study was to identify predictors of unsuppressed viral load among adult patients taking antiretroviral therapy at selected public and private health facilities of Adama town, East shewa zone, Ethiopia. METHODS: An unmatched case-control study was conducted from April 15 /2021 to May 20/2021. A total sample size of 347 patients consisting 116 cases and 231 controls was selected from electronic database among patients who started ART from September 2015 to August 2020. Data were collected using checklist from patient medical records and analyzed by SPSS. The association of dependent and independent variables was determined using multivariate analysis with 95% confidence interval and P - value in logistic regression model to identify independent predictors. RESULT: From the total 347 participants, 140 (40.3%) of them were males and 207 (59.7%) were females. In multivariate logistic regression, CD4 count < 100 [(AOR:1.22, 95% CI: 1.4-7.3)], CD4 100-200[(AOR: 2.58 95% CI: 1.06-8.28)], Fair Adherence [(AOR: 2.44, 95% CI: 1.67-4.82)], poor adherence [(AOR: 1.11, 95% CI: 1.7-6.73)], History of Cotrimoxazole Therapy (CPT) use and not used [(AOR: 2.60, 95% CI: 1.23-5.48)] and History of drug substitution [(AOR:. 361, 95% CI: .145-.897)] were independent predictors of unsuppressed viral load with the p-value less than 0.05. CONCLUSION AND COMMENDATION: In this study, Baseline CD4, adherence, History of CPT used and history of drug substitution was predictors of unsuppressed viral load. Monitoring immunological response through scheduled CD4 tests is essential to maintain immunity of the patients preventing diseases progression. Intensive adherence support and counseling should conclusively be provided through effective implementation of ART programs by providers would enhance viral suppression ensuring the quality of care and treatment.

Anopheles stephensi Mosquitoes as Vectors of Plasmodium vivax and falciparum, Horn of Africa, 2019.

Anopheles stephensi mosquitoes, efficient vectors in parts of Asia and Africa, were found in 75.3% of water sources surveyed and contributed to 80.9% of wild-caught Anopheles mosquitoes in Awash Sebat Kilo, Ethiopia. High susceptibility of these mosquitoes to Plasmodium falciparum and vivax infection presents a challenge for malaria control in the Horn of Africa.

Comparison of infectivity of Plasmodium vivax to wild-caught and laboratory-adapted (colonized) Anopheles arabiensis mosquitoes in Ethiopia.

BACKGROUND: Mosquito-feeding assays that assess transmission of Plasmodium from man-to-mosquito typically use laboratory mosquito colonies. The microbiome and genetic background of local mosquitoes may be different and influence Plasmodium transmission efficiency. In order to interpret transmission studies to the local epidemiology, it is therefore crucial to understand the relationship between infectivity in laboratory-adapted and local mosquitoes. METHODS: We assessed infectivity of Plasmodium vivax-infected patients from Adama, Ethiopia, using laboratory-adapted (colony) and wild-caught (wild) mosquitoes raised from larval collections in paired feeding experiments. Feeding assays used 4-6 day-old female Anopheles arabiensis mosquitoes after starvation for 12 h (colony) and 18 h (wild). Oocyst development was assessed microscopically 7 days post-feeding. Wild mosquitoes were identified morphologically and confirmed by genotyping. Asexual parasites and gametocytes were quantified in donor blood by microscopy. RESULTS: In 36 paired experiments (25 P. vivax infections and 11 co-infections with P. falciparum), feeding efficiency was higher in colony (median: 62.5%; interquartile range, IQR: 47.0-79.0%) compared to wild mosquitoes (median: 27.8%; IQR: 17.0-38.0%; Z = 5.02; P < 0.001). Plasmodium vivax from infectious individuals (51.6%, 16/31) infected a median of 55.0% (IQR: 6.7-85.7%; range: 5.5-96.7%; n = 14) of the colony and 52.7% (IQR: 20.0-80.0%; range: 3.2-95.0%; n = 14) of the wild mosquitoes. A strong association (ρ(16) = 0.819; P < 0.001) was observed between the proportion of infected wild and colony mosquitoes. A positive association was detected between microscopically detected gametocytes and the proportion of infected colony (ρ(31) = 0.452; P = 0.011) and wild (ρ(31) = 0.386; P = 0.032) mosquitoes. CONCLUSIONS: Infectivity assessments with colony and wild mosquitoes yielded similar infection results. This finding supports the use of colony mosquitoes for assessments of the infectious reservoir for malaria in this setting whilst acknowledging the importance of mosquito factors influencing sporogonic development of Plasmodium parasites.

The Relative Contribution of Symptomatic and Asymptomatic Plasmodium vivax and Plasmodium falciparum Infections to the Infectious Reservoir in a Low-Endemic Setting in Ethiopia.

Background: The majority of Plasmodium vivax and Plasmodium falciparum infections in low-endemic settings are asymptomatic. The relative contribution to the infectious reservoir of these infections compared to clinical malaria cases is currently unknown. Methods: We assessed infectivity of passively recruited symptomatic malaria patients (n = 41) and community-recruited asymptomatic individuals with microscopy-detected (n = 41) and polymerase chain reaction (PCR)-detected infections (n = 82) using membrane feeding assays with Anopheles arabiensis mosquitoes in Adama, Ethiopia. Malaria incidence and prevalence data were used to estimate the contributions of these populations to the infectious reservoir. Results: Overall, 34.9% (29/83) of P. vivax- and 15.1% (8/53) P. falciparum-infected individuals infected ≥1 mosquitoes. Mosquito infection rates were strongly correlated with asexual parasite density for P. vivax (ρ = 0.63; P < .001) but not for P. falciparum (ρ = 0.06; P = .770). Plasmodium vivax symptomatic infections were more infectious to mosquitoes (infecting 46.5% of mosquitoes, 307/660) compared to asymptomatic microscopy-detected (infecting 12.0% of mosquitoes, 80/667; P = .005) and PCR-detected infections (infecting 0.8% of mosquitoes, 6/744; P < .001). Adjusting for population prevalence, symptomatic, asymptomatic microscopy-detected, and PCR-detected infections were responsible for 8.0%, 76.2%, and 15.8% of the infectious reservoir for P. vivax, respectively. For P. falciparum, mosquito infections were sparser and also predominantly from asymptomatic infections. Conclusions: In this low-endemic setting aiming for malaria elimination, asymptomatic infections were highly prevalent and responsible for the majority of onward mosquito infections. The early identification and treatment of asymptomatic infections might accelerate elimination efforts.