Prevalence of malaria and associated risk factors among household members in South Ethiopia: a multi-site cross-sectional study.

BACKGROUND: Despite continuous prevention and control strategies in place, malaria remains a major public health problem in sub-Saharan Africa including Ethiopia. Moreover, prevalence of malaria differs in different geographical settings and epidemiological data were inadequate to assure disease status in the study area. This study was aimed to determine the prevalence of malaria and associated risk factors in selected rural kebeles in South Ethiopia. METHODS: A community-based cross-sectional study was conducted between February to June 2019 in eight malaria-endemic kebeles situated in four zones in South Ethiopia. Mult-stage sampling techniques were employed to select the study zones, districts, kebeles and households. Blood sample were collected from 1674 participants in 345 households by finger prick and smears were examined by microscopy. Sociodemographic data as well as risk factors for Plasmodium infection were collected using questionnaires. Bivariate and multivariate logistic regressions were used to analyse the data. RESULTS: The overall prevalence of malaria in the study localities was 4.5% (76/1674). The prevalence was varied among the study localities with high prevalence in Bashilo (14.6%; 33/226) followed by Mehal Korga (12.1%; 26/214). Plasmodium falciparum was the dominant parasite accounted for 65.8% (50/76), while Plasmodium vivax accounted 18.4% (14/76). Co-infection of P. falciparum and P. vivax was 15.8% (12/76). Among the three age groups prevalence was 7.8% (27/346) in age less than 5 years and 7.5% (40/531) in 5-14 years. The age groups > 14years were less likely infected with Plasmodium parasite (AOR = 0.14, 95% CI 0.02-0.82) than under five children. Non-febrile individuals 1638 (97.8%) were more likely to had Plasmodium infection (AOR = 28.4, 95% CI 011.4-70.6) than febrile 36 (2.2%). Individuals living proximity to mosquito breeding sites have higher Plasmodium infection (AOR = 6.17, 95% CI 2.66-14.3) than those at distant of breeding sites. CONCLUSIONS: Malaria remains a public health problem in the study localities. Thus, malaria prevention and control strategies targeting children, non-febrile cases and individuals living proximity to breeding sites are crucial to reduce malaria related morbidity and mortality.

Anopheles arabiensis continues to be the primary vector of Plasmodium falciparum after decades of malaria control in southwestern Ethiopia.

BACKGROUND: Investigating the species distribution and their role in malaria transmission is important as it varies from place to place and is highly needed to design interventions appropriate to the site. The current study aimed to investigate the Anopheles mosquito species distribution and their infection rate in southwestern Ethiopia. METHODS: The study was conducted in 14 malaria-endemic kebeles (the smallest administrative unit), which were situated in eight different malaria-endemic districts and four zones in southwestern Ethiopia. Ten per cent of households in each village were visited to collect adult mosquitoes using Centers for Disease Control and Prevention (CDC) light traps. The larval and pupal collection was done from breeding sites within the villages, and reared to adults. Female mosquitoes were morphologically identified. The head and thorax of adult Anopheles mosquitoes were tested for circumsporozoite proteins (CSPs) using ELISA. At the same time, legs, wings, and abdomen were used to identify sibling species using PCR targeting the rDNA intergenic spacers region for species typing of the Anopheles funestus group and the internal transcribed spacer 2 region genes for Anopheles gambiae complex. RESULTS: A total of 1445 Anopheles mosquitoes comprising eight species were collected. Of 813 An. gambiae complex tested by PCR, 785 (97%) were Anopheles arabiensis, and the remaining 28 (3%) were not amplified. There were 133 An. funestus group captured and tested to identify the species, of which 117 (88%) were positive for Anopheles parensis, and 15 (11%) were not amplified. A single specimen (1%) showed a band with a different base pair length from the known An. funestus group species. Sequencing revealed this was Anopheles sergentii. Among 1399 Anopheles tested for CSPs by ELISA, 5 (0.4%) An. arabiensis were positive for Plasmodium falciparum and a single (0.07%) was positive for Plasmodium vivax. CONCLUSIONS: Anopheles arabiensis continues to play the principal role in malaria transmission despite implementing indoor-based interventions for decades. Sequencing results suggest that An. sergentii was amplified by the An. funestus group primer, producing PCR amplicon size of different length. Therefore, relying solely on amplifying a specific gene of interest in grouping species could be misleading, as different species may share the same gene.

High human blood meal index of mosquitoes in Arba Minch town, southwest Ethiopia: an implication for urban mosquito-borne disease transmission.

Unplanned human population shifts in urban areas are expected to increase the prevalence of vector-borne diseases. This study aimed to investigate mosquito species composition, blood meal sources, and malaria vectors in an urban area. Indoor-resting adult mosquitoes were collected using Prokopack and host-seeking mosquitoes using Centers for Disease Control and Prevention light traps in Arba Minch town. Larval collection from artificial containers was done in those houses selected for adult mosquito collection. Anopheles adults collected and emerged from larvae were identified morphologically using a taxonomic key. ELISA was used to identify blood meal sources in freshly fed Anopheles and Culex mosquitoes, and CSP of Anopheles mosquitoes. A total of 16,756 female mosquitoes were collected. Of these, 93% (15,571) were Culex, 6% (1016) were Anopheles, and 1% (169) were Aedes mosquitoes. Out of the 130 adult mosquitoes that were raised from larvae collected from the containers, 20% were An. rhodesiensis, while the remaining 80% were Aedes mosquitoes. Out of 823 mosquitoes tested for blood meal origins, 86.3% (710/823) tested positive for human blood, 2.2% (18/823) tested positive for bovine blood, and 11.5% (95/823) were negative for human and bovine antibodies. Anopheles gambiae complex had a human blood meal index (HBI) of 50% (90/180; CI 42.3-57.5%) and a bovine blood meal index (BBI) of only 0.5% (95% CI 0.01-3.1%). Culex HBI was 96.7% (620/641), and its BBI index was 2.4% (15/641). While it was low (0.8%) in Culex, the proportion of An. gambiae complex with unidentified blood meal sources was 49.5% (95 CI% 41.9-56.9%). Among the 1016 Anopheles mosquitoes tested, a single An. gambiae complex (0.1%; 1/1016) was positive for P. vivax CSP. The high HBI indicates frequent contact between humans and vectors. To reduce human exposure, personal protection tools should be implemented.

Cattle feeding tendency of Anopheles mosquitoes and their infection rates in Aradum village, North Wollo, Ethiopia: an implication for animal-based malaria control strategies.

BACKGROUND: Surveillance of indoor and outdoor resting malaria vector populations is crucial to monitor possible changes in vector resting and feeding behaviours. This study was conducted to assess the resting behaviour, blood meal sources and circumsporozoite (CSP) rates of Anopheles mosquito in Aradum village, Northern Ethiopia. METHODS: Mosquito collection was conducted from September 2019 to February 2020 using clay pots (indoor and outdoor), pit shelter and pyrethrum spray catches (PSC). The species of Anopheles gambiae complex and Anopheles funestus group were identified using polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was done to determine CSP and blood meal sources of malaria vectors. RESULTS: A total of 775 female Anopheles mosquitoes were collected using the clay pot, PSC and pit shelter. Seven Anopheles mosquito species were identified morphologically, of which Anopheles demeilloni (593; 76.5%) was the dominant species followed by An. funestus group (73; 9.4%). Seventy-three An. funestus group screened by PCR, 91.8% (67/73) were identified as Anopheles leesoni and only 2.7% (2/73) were found to be Anopheles parensis. The molecular speciation of 71 An. gambiae complex confirmed 91.5% (65/71) of Anopheles arabiensis. The majority of Anopheles mosquitoes were collected from outdoor pit shelter (42.2%) followed by outdoor clay pots. The majority of the blood meal of An. demeilloni (57.5%; 161/280), An. funestus sensu lato 10 (43.5%) and An. gambiae (33.3%; 14/42) originated from bovine. None of the 364 Anopheles mosquitoes tested for Plasmodium falciparum and Plasmodium vivax sporozoite infections were positive. CONCLUSION: Since the Anopheles mosquitoes in the area prefer to bite cattle, it may be best to target them with an animal-based intervention. Clay pots could be an alternative tool for outdoor monitoring of malaria vectors in areas where pit shelter construction is not possible.

Malaria misdiagnosis in the routine health system in Arba Minch area district in southwest Ethiopia: an implication for malaria control and elimination.

BACKGROUND: Plasmodium falciparum and Plasmodium vivax are coendemic in Ethiopia, with different proportion in different settings. Microscopy is the diagnostic tool in Ethiopian health centres. Accurate species-specific diagnosis is vital for appropriate treatment of cases to interrupt its transmission. Therefore, this study assessed the status of species-specific misdiagnosis by microscope compared with polymerase chain reaction (PCR). METHODS: A health facility based cross-sectional study was conducted from November 2019 to January 2020 in Kolla Shelle Health centre, Arba Minch Zuria district. The study population were suspected malaria cases, who visited the health centre for a diagnosis and treatment. Consecutive microscopy positive cases as well as a sample of microscopically negative cases were included for molecular analysis by polymerase chain reaction (PCR). RESULTS: 254 microscopically negative and 193 microscopically positive malaria suspects were included. Of the 193 malaria positive cases, 46.1% [95% confidence interval (CI) 38.9-53.4] (89/193) were P. falciparum infection, 52.3% (95% CI 45.0-59.5) (101/193) were P. vivax infection, and 1.6% (3/193) had mixed infection of P. falciparum and P. vivax. Of the microscopically positive cases of P. falciparum, 3.4% (3/89) were P. vivax and 11.2% (10/89) were mixed infections with P. falciparum and P. vivax and a single case was negative molecularly. Similarly, of the microscopically positive P. vivax cases, 5.9% (6/101) were P. falciparum and 1% (1/101) was mixed infection. Single case was negative by molecular technique. Of the 254 microscopically negative cases, 0.8% were tested positive for P. falciparum and 2% for P. vivax by PCR. Considering molecular technique as a reference, the sensitivity of microscopy for detecting P. falciparum was 89.2% and for P. vivax, it was 91.2%. The specificity of microscopy for detecting P. falciparum was 96.1% and for P. vivax, it was 97.7%. However, the sensitivity of microscopy in detecting mixed infection of P. falciparum and P. vivax was low (8.3%). CONCLUSION: There were cases left untreated or inappropriately treated due to the species misidentification. Therefore, to minimize this problem, the gaps in the microscopic-based malaria diagnosis should be identified. It is recommended to regularly monitor the competency of malaria microscopists in the study area to improve species identification and diagnosis accuracy.

Effect of malaria prevention education on bed net utilization, incidence of malaria and treatment seeking among school-aged children in Southern Ethiopia; cluster randomized controlled trial.

BACKGROUND: School-aged children (SAC) have an increased risk to contract malaria and play a major role in its transmission dynamics. However, their malaria prevention experience is poor. Thus, the effect of malaria prevention education (MPE) on bed net utilization, treatment seeking from a health facility and cumulative incidence of malaria was evaluated in Southern Ethiopia. METHODS: A two arm cluster randomized controlled trial was conducted by recruiting 2038 SAC from 32 schools. Structured questionnaire was used to collect data on socio-demographic, economic, bed net ownership, bed net utilization, whether the participated child suffered from malaria and has got treatment from a health facility. Generalized mixed effect logistic regression using school as random variable was used to assess the effect of the intervention on the outcome variables. RESULTS: The ownership of bed net in households of the control and intervention schools was similar respectively with 84.6 and 88.6% (Crude Odds Ratio (COR): 1.5; 95%CI: 0.5-4.8). The percentage of SAC slept under the bed net the night before the survey was also similar (55.1% versus 54.0%); COR:1.04; 95%CI: 0.5-2.4). Bed net utilization was affected by household size to the bed net ratio ≤ 2 (Adjusted Odds Ratio (AOR) = 1.6; 95%CI:1.3-2.1), bed net utilization at baseline of the study (AOR = 2.3; 95%CI:1.5-3.6), and history of malaria attack in the last twelve months (AOR = 1.3; 95%CI:1.01-1.8). Reported cumulative incidence of malaria and treatment seeking from a health facility by SAC was similar between intervention and control arms: -2.1% (COR = 0.8; 95%CI: 0.5-1.5) and 9.6% (COR = 1.4; 95%CI: 0.4-4.3) respectively. The reported incidence of malaria was affected by altitude (AOR = 0.5; 95%CI: 0.3-0.8), low and medium wealth index (AOR = 0.7; 95%CI: 0.5-0.96 and AOR = 0.7; 95%CI: 0.5-0.98), adequate bed net number for household members (AOR = 0.7; 95%CI:0.5-0.9) and bed net utilization (AOR = 1.3; 95%CI:1.1-1.8). CONCLUSIONS: MPE had no significant effect on the use of malaria prevention measures considered, treatment seeking from a health facility and reported cumulative incidence of malaria though bed net use was associated with malaria incidence. Before organizing any health education program, sustainable implementation efforts have to be warranted especially in SAC, a neglected but relevant vulnerable and reservoirs. TRIAL REGISTRATION: Pan African Clinical Trials Registry PACTR202001837195738, registered 21/01/2020.

Cattle-related risk factors for malaria in southwest Ethiopia: a cross-sectional study.

BACKGROUND: Despite the low to moderate intensity of malaria transmission present in Ethiopia, malaria is still a leading public health problem. Current vector control interventions, principally long-lasting insecticidal nets and indoor residual spraying, when deployed alone or in combination, are insufficient to control the dominant vector species due to their exophagic and exophilic tendencies. Zooprophylaxis presents a potential supplementary vector control method for malaria; however, supporting evidence for its efficacy has been mixed. METHODS: To identify risk factors of malaria and to estimate the association between cattle and Anopheles vector abundance as well as malaria risk, a cross-sectional study was conducted in a village near Arba Minch, Ethiopia. Epidemiological surveys (households = 95, individuals = 463), mosquito collections using CDC light traps and a census of cattle and human populations were conducted. To capture environmental conditions, land cover and water bodies were mapped using satellite imagery. Risk factor analyses were performed through logistic, Poisson, negative binomial, and spatial weighted regression models. RESULTS: The only risk factor associated with self-reported malaria illness at an individual level was being a child aged 5 or under, where they had three times higher odds than adults. At the household level, variables associated with malaria vector abundance, especially those indoors, included socioeconomic status, the proportion of children in a household and cattle population density. CONCLUSIONS: Study results are limited by the low abundance of malaria vectors found and use of self-reported malaria incidence. Environmental factors together with a household's socioeconomic status and host availability played important roles in the risk of malaria infection in southwest Ethiopia. Cattle abundance in the form of higher cattle to human ratios may act as a protective factor against mosquito infestation and malaria risk. Humans should remain indoors to maximize potential protection against vectors and cattle kept outside of homes.

Implementation of a malaria prevention education intervention in Southern Ethiopia: a qualitative evaluation.

BACKGROUND: Though school-aged children (SAC) are at high risk of malaria, they are the ones that benefit the least from malaria prevention measures. A cluster randomized controlled trial was conducted to evaluate the effect of malaria prevention education (MPE) on insecticide-treated bed net (ITN) utilization and prompt diagnosis, reported incidence and treatment (PDAT) of malaria. Qualitative evaluation of the implementation of such interventions is vital to explain its effectiveness and will serve as guidance for future interventions. Therefore, this study aimed to evaluate the implementation of the MPE in southern Ethiopia.  METHODS: The trial was registered in Pan African Clinical Trials Registry (PACTR202001837195738) on 21/01/2020. A descriptive qualitative study using semi-structured interview with participants of the MPE was conducted in January 2020 and January 2021. The collected data were transcribed verbatim and analyzed thematically. The analysis of the data was supported by NVivo. RESULTS: The four themes identified after evaluation of MPE training were the setup of the training, challenges for the success of the training, anticipated challenges for practice as per the protocol and experienced immediate influences of the training. Participants appreciated the training: content covered, way of delivery and the mix of the participants. The context specific facilitators to bed net use were the collateral benefits of ITN and perceived at high risk of malaria while its barriers were quality and quantity of the bed nets, bed net associated discomforts, malaria health literacy and housing condition. Severeness of malaria symptoms and malaria health literacy were reported as both barriers and facilitators of the PDAT of malaria. The identified facilitators of PDAT of malaria were health professionals' attitude and exposure to MPE while its barriers were poverty, use of traditional medicine, health facility problems and Coronavirus Disease 2019 (COVID-19) pandemic. CONCLUSION: Low attendance of parents in the training was the major challenge for the success of MPE. National malaria program should ensure the access to malaria prevention measures; and future studies using increased frequency of the intervention embedded with monitoring adherence to the intervention protocol shall be conducted to improve the gains from existing malaria interventions.

Prevalence and associated risk factors of asymptomatic malaria and anaemia among school-aged children in Dara Mallo and Uba Debretsehay districts: results from baseline cluster randomized trial.

BACKGROUND: Despite the growing evidence that malaria and anaemia are two interlinked health problems of school-aged children (SAC) in developing countries, there is scarce information about malaria among SAC in Ethiopia. Moreover, anaemia-related studies were more concentrated in easily accessible areas. This study aimed to assess the prevalence of malaria and anaemia and corresponding risk factors among SAC in Dara Mallo and Uba Debretshay districts, in hard to reach areas, so as to inform appropriate integrated interventions for both diseases. METHODS: This study was part of baseline data collected for a cluster-randomized trial registered in Pan African Clinical Trials Registry (PACTR202001837195738). Data were collected from 2167 SAC and their households through face-to-face interview; malaria was diagnosed by using rapid diagnostic test (RDT); haemoglobin concentration was determined using hemoCue hb 301 and adjusted for altitude to determine anaemic status; helminth infections were determined by using kato-katz, and anthropometric measurements were made to determine nutritional status of children. Generalized mixed effects logistic regression model was used to assess the association between predictor variables and malaria and anaemia using school as a random variable. RESULTS: The overall prevalence of malaria was 1.62% (95% CI 1.15-2.27%) (35/2167). Of the 35 children positive for malaria, 20 (57.14%), 3 (8.57%) and 12 (34.29%) were due to Plasmodium falciparum, Plasmodium vivax and mixed infections of P. falciparum and P. vivax, respectively. Malaria was significantly lower among children from literate household head (Adjusted OR = 0.38; 95% CI 0.15-0.95) and residence house located at an altitude range above 1100 masl (AOR = 0.40; 95% CI 0.17-0.94). The prevalence of anaemia was 22.00% (95% CI 20.3-23.8%) (477/2167) and was significantly reduced by eating legumes, nuts or seed group of food in their 24-h dietary diversity recall (AOR = 0.64; 95% CI 0.41-0.99). CONCLUSIONS: The prevalence of malaria was low and unevenly distributed per school while the overall prevalence of anaemia was moderate. It is important to implement integrated interventions targeting both malaria and anaemia, with special emphasis given to children from illiterate households and living at an altitude below 1100 masl. The micronutrient content of locally grown legumes should be further investigated to recommend specific interventions to overcome anaemia.

Long-lasting insecticide-treated bed net ownership, utilization and associated factors among school-age children in Dara Mallo and Uba Debretsehay districts, Southern Ethiopia.

BACKGROUND: Malaria is one of the major causes of morbidity and mortality among school-age children (SAC) in sub-Saharan Africa. SAC account for more than 60% of the reservoir of malaria transmission, but they are given less emphasis in prioritizing malaria prevention interventions. This study was aimed at assessing the ownership of long-lasting insecticide treated bed nets (LLINs), its utilization and factors associated with ownership of LLINs by households and LLINs utilization among SAC in malaria-prone areas of Dara Mallo and Uba Debretsehay districts in Southern Ethiopia, October to December 2019. METHODS: This study is part of a baseline assessment in a cluster-randomized controlled trial. The data was collected through interview and observation, following a structured questionnaire, of 2261 SAC households. Univariable and multivariable multilevel logistic regressions were used to assess the association between LLINs ownership and utilization and potential predictor variables. Odds ratio (OR) and corresponding 95% confidence interval (CI) were used to determine the strength and statistical significance of association. RESULTS: The ownership of at least one LLIN by households of SAC was about 19.3% (95% CI 17.7-21.0%) but only 10.3% % (95% CI 7.7-13.7%) of these households had adequate access of bed nets to the household members. Ownership of bed net was marginally affected by living in semi-urban area (adjusted OR = 2.6; 95% CI 1.0-6.9) and occupational status of the household head being a civil servant (adjusted OR = 2.7; 95% CI 0.9-7.9). About 7.8% (95% CI 6.7-10.0%) of all SAC participated in the study and 40.4% (95% CI 57.4-66.7%) of children in households owning at least one LLIN passed the previous night under LLIN. LLIN utilization by SAC conditional to presence of at least one net in the household was significantly correlated with education level of mother above grade 6 (adjusted OR = 3.4; 95% CI 1.3-9.3) and the household size to bed net ratio less than or equal to 2 (adjusted OR = 20.7; 95% CI 4.7-132.5). CONCLUSION: Ownership of bed net was lower than universal coverage of at least one bed net for two individuals. It is important to monitor replacement needs and educate mothers with low education level with their SAC on the benefit of consistent utilization of bed nets.

The effect of ivermectin® on fertility, fecundity and mortality of Anopheles arabiensis fed on treated men in Ethiopia.

BACKGROUND: Insecticide resistance is a growing threat to malaria vector control. Ivermectin, either administered to humans or animals, may represent an alternate strategy to reduce resistant mosquito populations. The aim of this study was to assess the residual or delayed effect of administering a single oral dose of ivermectin to humans on the survival, fecundity and fertility of Anopheles arabiensis in Ethiopia. METHODS: Six male volunteers aged 25-40 years (weight range 64-72 kg) were recruited; four of them received a recommended single oral dose of 12 mg ivermectin and the other two individuals were untreated controls. A fully susceptible insectary colony of An. arabiensis was fed on treated and control participants at 1, 4, 7, 10 and 13 days post ivermectin-administration. Daily mosquito mortality was recorded for 5 days. An. arabiensis fecundity and fertility were measured from day 7 post treatment, by dissection to examine the number of eggs per mosquito, and by observing larval hatching rates, respectively. RESULTS: Ivermectin treatment induced significantly higher An. arabiensis mortality on days 1 and 4, compared to untreated controls (p = 0.02 and p < 0.001, respectively). However, this effect had declined by day 7, with no significant difference in mortality between treated and control groups (p = 0.06). The mean survival time of mosquitoes fed on day 1 was 2.1 days, while those fed on day 4 survived 4.0 days. Mosquitoes fed on the treatment group at day 7 and 10 produced significantly lower numbers of eggs compared to the untreated controls (p < 0.001 and p = 0.04, respectively). An. arabiensis fed on day 7 on treated men also had lower larval hatching rates than mosquitoes fed on days 10 and 13 (p = 0.003 and p = 0.001, respectively). CONCLUSION: A single oral dose of ivermectin given to humans can induce mortality and reduce survivorship of An. arabiensis for 7 days after treatment. Ivermectin also had a delayed effect on fecundity of An. arabiensis that took bloodmeals from treated individuals on day 7 and 10. Additional studies are warranted using wild, insecticide-resistant mosquito populations, to confirm findings and a phase III evaluation among community members in Ethiopia is needed to determine the impact of ivermectin on malaria transmission.

Wall-type and indoor residual spraying application quality affect the residual efficacy of indoor residual spray against wild malaria vector in southwest Ethiopia.

BACKGROUND: Residual efficacy of indoor residual spray may vary with different spray quality and wall surfaces types. This study evaluated the impact of spray quality and wall surface types on residual efficacy of propoxur against wild Anopheles gambiae sensu lato (s.l.) in southwest Ethiopia. METHODS: Thirty houses of different mud wall surfaces (10 smooth, 10 rough, 10 painted) were selected and randomly allocated into routine and standard spray. The routine spray was conducted by district health office as usual, while the standard spray was done by strictly following guidelines. Three control houses were selected from unsprayed nearby semi-urban. Wild An. gambiae s.l. were used for wall bioassay tests. Two-way mixed model analysis of variance was used to analyse the data. The mean variation between wall and spray types was compared by post hoc analysis of IBM SPSS version 20. RESULTS: On standard spray, knockdown rate was 95.3% on painted, 82% on smooth and 72.5% on rough surface at week 17 of post-spray, whereas on routine spray it was 82.7% on painted, 48.7% on smooth and 60% on rough surface. On standard spray, mortality rate of An. gambiae s.l. was 99.3% on painted surface, 90% on smooth and 80% on rough surface. On routine spray, it was 89.3% on painted, 61.3% on smooth and 65% on rough surface at week 17 of post-spray. The painted wall surface showed the highest knockdown rate (86.4-100%) on standard and (73.8-91.5%) routine spray; mortality rate was more than 80% on both spray types during the 17 weeks of follow-up regardless of spray types. The lower mortality rate and residual effect was observed on routine smooth and rough wall surfaces. The residual efficacy of propoxur was > 80% at week 17 on standard spray regardless of the wall types and it was < 80% on routine spray except painted wall surface. CONCLUSION: The painted wall surface and standard spray showed better residual efficacy. Therefore, it is recommended to consider the wall surface available in the community to estimate the residual lifespan of the insecticide, and strictly to follow the spray guideline to improve the effectiveness of indoor residual spray.

Status of Bactrocera invadens (Diptera: Tephritidae) in mango-producing areas of Arba Minch, southwestern Ethiopia.

Bactrocera invadens, the Asian fruit fly, was first reported in Kenya in 2003, and it spread fast to most tropical countries in Africa. To our knowledge, there is no detailed data on the fruit damage and status of fruit flies in Arba Minch and elsewhere in Ethiopia. Hence, information on the species composition and pest status of the fruit fly species is urgent to plan management strategies in the area. Fruit flies were captured using male parapheromone-baited traps. Matured mango (Mangifera indica) fruits were collected from randomly selected mango trees and incubated individually in cages (15 by 15 by 15 cm) with sandy soil. B. invadens was the predominant (96%; 952 of 992) captured species and the only fruit fly species emerging from mango fruits incubated in the laboratory. The mean number of adult B. invadens emerging per mango fruit was 35.25, indicating that the species is the most devastating mango fruit fly in the area. The loss due to this species would be serious if no management strategies are implemented.

Administration of ivermectin to cattle induced mortality, reduced fecundity and survivorship of Anopheles arabiensis in Ethiopia: an implication for expansion of vector control toolbox.

BACKGROUND: Although many countries have shown interest in eliminating malaria, approaches that complement existing vector control interventions are needed because existing methods have been scaled up but malaria still persists. Therefore, the effect of ivermectin administration to cattle was evaluated for its effect on mortality, survivorship and mortality of laboratory reared Anopheles arabiensis. METHODS: Three calves were randomly selected and injected with ivermectin at a therapeutic dose of 0.2 mg/kg, while the other two calves received no treatment and served as controls. Five tents were constructed for the trial. Calves were housed in tents (one per tent) and then 30 starved female An. arabiensis were introduced into each tent. Only fully engorged females were collected from each tent and placed in different mosquito cages to monitor their mortality, survival and fecundity. Data analysis was done using SPSS version 16. RESULTS: During the follow-up period (until day 21), ivermectin induced significantly higher mortality when compared to controls. It resulted in an average 24-h mortality rate of 81.6% against An. arabiensis on the first day following treatment. 100% An. arabiensis that fed on ivermectin-treated calves on the first day after treatment died within four days. Egg production rate of An. arabiensis that fed on ivermectin-treated calves was significantly lower compared to controls (F = 768.7, P < 0.001). CONCLUSION: In conclusion, ivermectin induced mortality, reduced fecundity and survivorship of laboratory maintained An. arabiensis. Further study is recommended using a wild mosquito population. Moreover, mass ivermectin administration to domestic animals could be recommended to supplement the existing indoor based interventions.

Spatiotemporal distribution and bionomics of Anopheles stephensi in different eco-epidemiological settings in Ethiopia.

BACKGROUND: Malaria is a major public health concern in Ethiopia, and its incidence could worsen with the spread of the invasive mosquito species Anopheles stephensi in the country. This study aimed to provide updates on the distribution of An. stephensi and likely household exposure in Ethiopia. METHODS: Entomological surveillance was performed in 26 urban settings in Ethiopia from 2021 to 2023. A kilometer-by-kilometer quadrant was established per town, and approximately 20 structures per quadrant were surveyed every 3 months. Additional extensive sampling was conducted in 50 randomly selected structures in four urban centers in 2022 and 2023 to assess households' exposure to An. stephensi. Prokopack aspirators and CDC light traps were used to collect adult mosquitoes, and standard dippers were used to collect immature stages. The collected mosquitoes were identified to species level by morphological keys and molecular methods. PCR assays were used to assess Plasmodium infection and mosquito blood meal source. RESULTS: Catches of adult An. stephensi were generally low (mean: 0.15 per trap), with eight positive sites among the 26 surveyed. This mosquito species was reported for the first time in Assosa, western Ethiopia. Anopheles stephensi was the predominant species in four of the eight positive sites, accounting for 75-100% relative abundance of the adult Anopheles catches. Household-level exposure, defined as the percentage of households with a peridomestic presence of An. stephensi, ranged from 18% in Metehara to 30% in Danan. Anopheles arabiensis was the predominant species in 20 of the 26 sites, accounting for 42.9-100% of the Anopheles catches. Bovine blood index, ovine blood index and human blood index values were 69.2%, 32.3% and 24.6%, respectively, for An. stephensi, and 65.4%, 46.7% and 35.8%, respectively, for An. arabiensis. None of the 197 An. stephensi mosquitoes assayed tested positive for Plasmodium sporozoite, while of the 1434 An. arabiensis mosquitoes assayed, 62 were positive for Plasmodium (10 for P. falciparum and 52 for P. vivax). CONCLUSIONS: This study shows that the geographical range of An. stephensi has expanded to western Ethiopia. Strongly zoophagic behavior coupled with low adult catches might explain the absence of Plasmodium infection. The level of household exposure to An. stephensi in this study varied across positive sites. Further research is needed to better understand the bionomics and contribution of An. stephensi to malaria transmission.

The effect of screening doors and windows on indoor density of Anopheles arabiensis in south-west Ethiopia: a randomized trial.

BACKGROUND: Screening of houses might have impact on density of indoor host-seeking Anopheles mosquitoes. A randomized trial of screening windows and doors with metal mesh, and closing openings on eves and walls by mud was conducted to assess if reduce indoor densities of biting mosquitoes. METHODS: Mosquitoes were collected in forty houses using Centers for Diseases Control and Prevention (CDC) light traps biweekly in March and April 2011. A randomization of houses into control and intervention groups was done based on the baseline data. Windows and doors of 20 houses were screened by metal mesh, and openings on the walls and eves closed by mud and the rest 20 houses were used as control group. Mosquitoes were collected biweekly in October and November 2011 from both control and intervention houses. A Generalized Estimating Equations (GEE) with a negative binomial error distribution was used to account for over dispersion of Anopheles arabiensis and culicine counts and repeated catches made in the same house. RESULTS: Screening doors and windows, and closing openings on eves and wall by mud reduced the overall indoor densities of An. arabiensis by 40%. The effect of screenings pronounced on unfed An. arabiensis by resulting 42% reduction in houses with interventions. The total costs for screening windows and doors, and to close openings on the eves and walls by mud was 7.34 USD per house. CONCLUSION: Screening houses reduced indoor density of An. arabiensis, and it was cheap and can easily incorporated into malaria vector strategies by local communities, but improving doors and windows fitness for screening should be considered during house construction to increase the efficacy of screenings.

A dynamic model of some malaria-transmitting anopheline mosquitoes of the Afrotropical region. II. Validation of species distribution and seasonal variations.

BACKGROUND: The first part of this study aimed to develop a model for Anopheles gambiae s.l. with separate parametrization schemes for Anopheles gambiae s.s. and Anopheles arabiensis. The characterizations were constructed based on literature from the past decades. This part of the study is focusing on the model's ability to separate the mean state of the two species of the An. gambiae complex in Africa. The model is also evaluated with respect to capturing the temporal variability of An. arabiensis in Ethiopia. Before conclusions and guidance based on models can be made, models need to be validated. METHODS: The model used in this paper is described in part one (Malaria Journal 2013, 12:28). For the validation of the model, a data base of 5,935 points on the presence of An. gambiae s.s. and An. arabiensis was constructed. An additional 992 points were collected on the presence An. gambiae s.l.. These data were used to assess if the model could recreate the spatial distribution of the two species. The dataset is made available in the public domain. This is followed by a case study from Madagascar where the model's ability to recreate the relative fraction of each species is investigated. In the last section the model's ability to reproduce the temporal variability of An. arabiensis in Ethiopia is tested. The model was compared with data from four papers, and one field survey covering two years. RESULTS: Overall, the model has a realistic representation of seasonal and year to year variability in mosquito densities in Ethiopia. The model is also able to describe the distribution of An. gambiae s.s. and An. arabiensis in sub-Saharan Africa. This implies this model can be used for seasonal and long term predictions of changes in the burden of malaria. Before models can be used to improving human health, or guide which interventions are to be applied where, there is a need to understand the system of interest. Validation is an important part of this process. It is also found that one of the main mechanisms separating An. gambiae s.s. and An. arabiensis is the availability of hosts; humans and cattle. Climate play a secondary, but still important, role.

Plasmodium falciparum remains the dominant parasite affecting children despite decades of implementing vector control in two villages of Wolaita Zone, Southwest Ethiopia.

Background: Malaria is still a significant public health concern, and its prevention and control measures have different impacts in different areas. This study assesses the prevalence of malaria and the effectiveness of routine malaria control programmes such as indoor residual spray (IRS) in two Ethiopian villages. Methods: The Kebeles (villages) were purposefully selected based on their malaria prevalence rates. A parasitology survey was conducted in Fango-Gelchecha pre- and post-IRS implementation, whereas in Shochora-Abela it was only conducted post-IRS implementation. The IRS was implemented as part of the routine malaria control programme in August 2017. Every fourth house from the village registration list was systematically selected, resulting in a sample of 300 households per village. A total of 3,075 individuals were enrolled for malaria testing using microscopy. Results: After three to four months of application of IRS in August 2017, 59 malaria cases were confirmed, resulting in an overall prevalence of 1.9% (95% CI: 1.5-2.5). Of the positive cases, 18 cases (0.59%: 95% CI: 1.3-1.8) were from Shochora-Abela village, and 41 cases (1.33%: 95% CI: 1.1-1.3) were from Fango-Gelchecha. About age categories, the prevalence of malaria was 10.1% (95% CI: 5.9-15.9) among children under five, 4.7% (95% CI: 3.3-6.4) in children aged 5-14, and only 0.32% (95% CI: 0.13-0.67) in the age group 15 and above. Overall, P. falciparum was the dominant malaria parasite, accounting for 69.5% (95% CI: 56.1-80.8), while P. vivax malaria accounted for 30.5% (95% CI: 19.2-43.8). The malaria prevalence in Fango-Gelchecha village was 3.1% (95% CI: 2.3-4.0) before IRS and 2.6% (95% CI: 1.8-3.5) after IRS application. In the village of Shochora-Abela, the prevalence of malaria post-IRS was 1.2% (95% CI: 0.7-1.9), but the prevalence prior to IRS was not evaluated. Conclusions: Plasmodium falciparum is the predominant parasite in the villages, mainly affecting children under five. Therefore, protecting young children should be the top priority for reducing infection burdens.

Anopheles mosquito diversity, entomological indicators of malaria transmission and challenges of morphological identification in southwestern Ethiopia.

BACKGROUND: A number of Anopheles species play either a primary or secondary role in malaria transmission. This necessitates understanding the species composition, bionomics, and behaviors of malaria mosquitoes in a particular geographic area, which is relevant to design and implement tailored intervention tools. This study aimed to assess the species composition, sporozoite infection rate, and blood meal origins of malaria mosquitoes in two malaria-endemic villages of Boreda district in Gamo Zone, southwest Ethiopia. METHODS: Thirty houses, 20 for Center for Disease Control and Prevention (CDC) light traps and 10 for Pyrethrum Spray Catches (PSC) were randomly selected for bimonthly mosquito collection from October 2019 to February 2020. An enzyme-linked immunosorbent assay (ELISA) was carried out to detect the blood meal origins and circumsporozoite proteins (CSPs). The entomological inoculation rate (EIR) was calculated by multiplying the sporozoite and human biting rates from PSCs. Anopheles gambiae complex and An. funestus group samples were further identified to species by the polymerase chain reaction (PCR). Anopheles species with some morphological similarity with An. gambiae complex or An. funestus group were tested using the primers of the two species complexes. RESULTS: A total of 14 Anopheles species were documented, of which An. demeilloni was found to be the dominant species. An. arabiensis was found to be positive for P. falciparum CSP with the overall CSP rate of 0.53% (1/190: 95% CI 0.01-2.9). The overall estimated P. falciparum EIR of An. arabiensis from PSC was 1.5 infectious bites/person/5 months. Of the 145 freshly fed Anopheles mosquitoes tested for blood meal sources, 57.9% (84/145) had bovine blood meal, 15.2% (22/145) had human blood meal origin alone, and 16.5% (24/145) had a mixed blood meal origin of human and bovine. Anopheles demeilloni were more likely to feed on blood meals of bovine origin (102/126 = 80.9%), while An. arabiensis were more likely to have blood meals of human origin. Eleven samples (2.6%; 11/420) were morphologically categorized as An. demeilloni, but it has been identified as An. leesoni (the only An. funestus group identified in the area) by PCR, though it requires additional verification by sequencing, because different species genes may have amplified for these species specific primers. Similarly, a small number of An. arabiensis were morphologically identified as An. salbaii, An. maculipalpis and An. fuscivenosus. CONCLUSIONS AND RECOMMENDATIONS: In spite of the wide variety of Anopheles mosquito species, An. arabiensis dominates indoor malaria transmission, necessitating additional interventions targeting this species. In addition, increasing entomological knowledge may make morphological identification less difficult.

Infectivity of symptomatic Plasmodium vivax cases to different generations of wild-caught and laboratory-adapted Anopheles arabiensis using a membrane feeding assay, Ethiopia.

When measuring human to mosquito transmission of Plasmodium spp., laboratory-adapted (colony) mosquitoes can be utilized. To connect transmission studies to the local epidemiology, it can be important to comprehend the relationship between infectivity in laboratory-adapted (colony) and wild-caught (wild) mosquitoes of the same species. Microscopically confirmed Plasmodium vivax cases were recruited from health facilities in Arba Minch town, and a nested polymerase chain reaction (nPCR) was used for subsequent confirmation. We performed paired membrane-feeding assays using colony An. arabiensis and three generations of wild origin An. arabiensis. Anopheles arabiensis aged 3-6 days were fed after being starved for 8-14 h. Microscopically, the oocyst development was evaluated at day 7 after feeding. Circumsporozoite proteins (CSPs) assay was carried out by enzyme-linked immunosorbent assay (ELISA). In 19 paired feeding experiments, the feeding efficiency was more than doubled in colony (median: 62.5%; interquartile range, IQR: 35-78%) than in wild mosquitoes (median: 28.5%; IQR: 17.5-40%; P < 0.001). Among the 19 P. vivax gametocyte-positive blood samples, 63.2% (n = 12) were infective to wild An. arabiensis and 73.7% (n = 14) were infective to colony An. arabiensis. The median infection rate was twice as high (26%) in the colony than in the wild (13%) An. arabiensis, although the difference was marginally insignificant (P = 0.06). Although the observed difference was not statistically significant (P = 0.19), the median number of oocysts per midgut was more than twice as high (17.8/midgut) in colony than in wild (7.2/midgut) An. arabiensis. The median feeding efficiency was 26.5% (IQR: 18-37%) in F1, 29.3% (IQR: 28-40%) in F2 and 31.2% (IQR: 30-37%) in F3 generations of wild An. arabiensis. Also, no significant difference was observed in oocyst infection rate and load between generations of wild An. arabiensis. CSP rate of P. vivax was 3.1% (3/97; 95% CI: 0.6-8.8%) in wild and 3.6% (3/84; 95% CI: 0.7-10.1%) in colony An. arabiensis. The results of the present study revealed that oocyst infection and load/midgut, and CSP rate were roughly comparable, indicating that colony mosquitoes can be employed for infectivity studies, while larger sample sizes may be necessary in future studies.