Meshesha Balkew

Insecticide resistance in Anopheles arabiensis (Diptera: Culicidae) from villages in central, northern and south west Ethiopia and detection of kdr mutation

BackgroundAnopheles arabiensis is the major vector of malaria in Ethiopia. Malaria vector control in Ethiopia is based on selective indoor residual spraying using DDT, distribution of long lasting insecticide treated nets and environmental management of larval breeding habitats. DDT and pyrethroid insecticides are neurotoxins and have a similar mode of action on the sodium ion channel of insects. It was therefore necessary to verify the insecticide susceptibility status of An. arabiensis, to better understand the status of cross-resistance between DDT and the pyrethroids in this species as well as to detect a resistant gene.MethodsStandard WHO insecticide susceptibility tests were conducted on adults reared from larval and pupal collections from breeding sites at three villages namely: Sodere in the Rift Valley, Gorgora in the north and Ghibe River Valley in the south west of Ethiopia. The occurrence of cross-resistance between pyrethroids and DDT was determined using a DDT selected laboratory colony originally collected from Gorgora. Phenotypically characterized mosquitoes were tested for the presence of knockdown resistance (kdr) alleles using the standard polymerase chain reaction assay.ResultsAll An. gambiae s.l. specimens assayed by PCR were identified as An. arabiensis. The knockdown and mortality results showed An. arabiensis resistance to DDT in all villages, resistance to deltamethrin and permethrin in the Ghibe River Valley and permethrin resistance in Gorgora. Bioassay susceptibility tests also indicated the presence of cross-resistance between DDT and permethrin, but not between DDT and deltamethrin. The knockdown resistance (kdr) mutation of leucine to phenylalanine in the sodium ion channel gene was detected in populations from Gorgora and the Ghibe River Valley.ConclusionSince An. arabiensis shows high levels of resistance to DDT in all villages tested and varying pyrethroid resistance in Gorgora and the Ghibe River valley, precautionary measures should be taken in future vector control operations. Moreover, the status of resistance in other locations in Ethiopia and the spread of resistant gene (s) should be investigated.

The isolation of Leishmania tropica and L. aethiopica from Phlebotomus (Paraphlebotomus) species (Diptera: Psychodidae) in the Awash Valley, northeastern Ethiopia.

In a survey of Leishmania infections in phlebotomine sandflies in a highly suspected focus of leishmaniasis in the Awash Valley (northeastern Ethiopia) between January 1994 and August 1997, a total of 3307 females of 11 Phlebotomus species (P. orientalis, P. fantalensis, P. saevus, P. sergenti, P. gemetchi, P. alexandri, P. bergeroti, P. duboscqi, P. arabicus, P. martini, and P. rodhaini) were dissected. Promastigotes were detected in 17 females of three species (11 P. saevus, 4 P. sergenti and 2 P. arabicus). Of these, only two P. saevus (one from Upper Awash and one from Middle Awash) and three P. sergenti (from Upper Awash) positives were successfully isolated in culture and were typed by isoenzyme analysis. Four isolates (two each from P. saevus and P. sergenti) were identified as new zymodemes (Z) of L. tropica and one isolate from P. sergenti was typed as a new zymodeme of L. aethiopica. This is the first finding of natural infections of P. saevus and P. arabicus and the first evidence for the former to be a vector of L. tropica. This is also the first time P. sergenti has been implicated in L. tropica transmission in Ethiopia; the isolation of L. aethiopica from a Paraphlebotomus species (P. sergenti) is also a new record. The possible presence of human cutaneous leishmaniasis (L. tropica and L. aethiopica), and wild reservoir host(s) of the parasites, especially rock hyrax (Procavia capensis) in the Upper and Middle Awash Valley remain to be determined.

Further studies on the phlebotomine sandflies of the kala-azar endemic lowlands of Humera-Metema (north-west Ethiopia) with observations on their natural blood meal sources

BackgroundVisceral leishmaniasis (VL) has been known to exist in northwest Ethiopia (Humera-Metema lowlands) since the early 1970s associated with large scale agricultural development activities, often resulting in outbreaks. The latest outbreak of the disease that has started around 1995 in both regions, has led to the present preliminary entomological surveys (1996-2005) the results of which are reported here. Sandflies were collected using CDC light traps and Phlebotomus females were dissected for Leishmania detection and isolation; freshly fed Phlebotomus females collected were subsequently tested for their blood meal sources using ELISA. All Phlebotomus collections were identified to species.ResultsDuring the surveys (1996-2005), a total of 1963 sandflies of six Phlebotomus species (P. orientalis, P. papatasi, P. bergeroti, P. duboscqi, P. rodhaini and P. alexandri) were recorded from the study areas: the predominant species was P. orientalis in both localities. None of the total 618 P. orientalis females dissected (506 from Metema and 112 from Humera), nor the total 114 females of four other species dissected (P. papatasi, P. duboscqi, P. bergeroti and P. rodhaini) was infected with Leishmania promastigotes. ELISA-based blood meal analysis of 273 fresh fed P. orientalis females collected from Metema revealed a remarkably high bovine blood feeds (92%) with only 2.2% of human blood feeds.ConclusionBased on abundance and other circumstantial evidences (its proven role in Sudan), P. orientalis is the most likely vector of VL in northwest Ethiopia, pending further clarifications. The zoophagic feeding behaviour of P. orientalis detected in the present study could have epidemiological significance, but more investigations are required in this and other behavioural characteristics towards appropriate management of the vector.

Blood meal origins and insecticide susceptibility of Anopheles arabiensis from Chano in South-West Ethiopia

BackgroundAnopheles arabiensis, the main malaria vector in Ethiopia, shows both anthropophilic and zoophilic behaviours. Insecticide resistance is increasing, and alternative methods of vector control are needed. The objectives of this study were to determine the blood meal origins and the susceptibility to insecticides of An. arabiensis from Chano village near Arba Minch in South-West Ethiopia.MethodsBlood meal sources of anopheline mosquitoes collected using Centers for Disease Control and Prevention (CDC) light traps and pyrethrum spray catches (PSC) from human dwellings, and hand-held mouth aspirators from outdoor pit shelters were analysed using a direct enzyme-linked-immunosorbent assay (ELISA). The susceptibility of An. arabiensis to pyrethroid insecticides (alphacypermethrin, lambdacyhalothrin, deltamethrin, and cyfluthrin) and DDT was assessed using females reared from larval and pupal collections from natural breeding sites.ResultsThe blood meal origins of 2967 freshly fed Anopheles mosquitoes were determined. An. arabiensis was the predominant species (75%), and it fed mainly on cattle. The densities of both freshly fed An. arabiensis and those fed on human blood followed similar seasonal patterns. The overall human blood index (HBI) of An. arabiensis, including mixed blood meals, was 44% and the bovine blood index (BBI) was 69%. The HBI of An. arabiensis from CDC light trap collections was 75% and this was higher than those for PSC (38%) and outdoor pit shelter collections (13%), while the BBI was 65% for PSC, 68% for outdoor pit shelters and 72% for CDC light traps. More freshly fed and human blood-fed An. arabiensis were sampled from houses close to the shore of Lake Abaya (the major breeding site).A high proportion of An. arabiensis was resistant to the pyrethroid insecticides, with a mortality rate of 56% for lambdacyhalothrin, 50% for cyfluthrin and alphacypermethrin, 47% for deltamethrin, and 10% for DDT.ConclusionAnopheles arabiensis is the predominant species of anopheline mosquito in this region, and cattle are the main source of its blood meals. The greater tendency of this species to feed on cattle justifies the application of insecticides on cattle to control it. However, An. arabiensis has already developed resistance to the available pyrethroid insecticides, and alternative insecticides are needed for malaria vector control.

Utilization, retention and bio-efficacy studies of PermaNet® in selected villages in Buie and Fentalie districts of Ethiopia

BackgroundMalaria remains a major public health problem in Ethiopia. Pyrethroid-treated mosquito nets are one of the major tools available for the prevention and control of malaria transmission. PermaNet® is a long-lasting insecticide-treated net (LLIN) recommended by WHO for malaria control.ObjectiveThe objective of the study was to assess utilization and retention of PermaNet® nets distributed for malaria control in Buie and Fentalie districts and monitor the bio-efficacy of the nets using the WHO cone bioassay test procedures.MethodsA cross sectional study was carried out by interviewing household heads or their representative in Buie and Fentalie districts. The two districts were selected based on a priori knowledge of variations on ethnic background and housing construction. Clusters of houses were chosen within each of the study villages for selection of households. 20 households that had received one or more PermaNet® nets were chosen randomly from the clusters in each village. A total of eight used PermaNet® nets were collected for the bio-efficacy test. The bio-efficacy of PermaNet® nets was monitored according to the standard WHO procedures using a susceptible colony of Anopheles arabiensis to deltamethrin.ResultsA total of 119 household heads were interviewed during the study. The retention rate of nets that were distributed in 2005 and 2006 season was 72%. A total of 62.2% of the interviewees claimed children under five years of age slept under LLIN, while only 50.7% of the nets were observed to be hanged inside houses when used as a proxy indicator of usage of LLIN. For the bio-efficacy test the mean knock-down was 94% and 100%, while the mean mortality rate observed after 24 hr holding period was 72.2% and 67% for Buie and Fentalie districts respectively.ConclusionThe study revealed a moderately high retention of PermaNet® in the study villages and effectiveness of the nets when tested according to the standard WHO procedure.

Laboratory and field evaluation of neem (Azadirachta indica A. Juss) and Chinaberry (Melia azedarach L.) oils as repellents against Phlebotomus orientalis and P. bergeroti (Diptera: Psychodidae) in Ethiopia

The study evaluated the efficacy of neem (Azadirachta indica A. Juss.) and Chinaberry (Melia azedarach L.) seed oils as repellents against laboratory and field populations of some sandflies in Ethiopia. In the laboratory, concentrations of 2% and 5% neem oil in coconut oil tested against Phlebotomus orientalis (vector of visceral leishmaniasis) provided 96.28% (95% CI=95.60-96.97) protection up to a mean time of 7h and 20 min and 98.26% (95% CI=93.46-104. 07) protection up to 9h, respectively. Similarly, M. azedarach oil at 2% concentration produced 95.13% (95% CI=90.74-99.52) protection for the same duration (7h and 20 min), while the 5% oil gave 96.20 (95% CI=86.98-105.41) protection for 8h and 20 min against the same species with no significant difference in percentage protection between the two oils at 2% and 5% concentrations. In the field tests with only neem oil (A. indica) against field populations of P. orientalis and P. bergeroti, similar high level of repellencies were recorded with about the same duration of protection. Application of both neem and Chinaberry oils can be safe and low-cost means of personal protection against sandfly bites in endemic areas of Ethiopia, if the community is advised and encouraged to grow the plants abundantly.

A zoonotic focus of cutaneous leishmaniasis in Addis Ababa, Ethiopia

BackgroundCutaneous leishmaniasis (CL) is endemic in the highlands of Ethiopia, and almost always caused by Leishmania aethiopica. Hitherto, Addis Ababa (the capital city of Ethiopia) was not considered endemic for CL, mainly due to absence of epidemiological and field ecological studies. This report summarizes the preliminary epidemiological investigation that proved the existence of active transmission in southeastern Addis Ababa.ResultsActive case finding surveys were conducted in 3 localities, Saris, Kality, and Akaki, which are found in and around Bulbula-Akaki river gorges. During the surveys conducted in January 2005 - May 2006, a total of 35 cases with 9 active and 26 healed skin lesions were identified. Eighteen of the cases (51.4%) were found in Saris; while 10 (28.6%) and 7 (20%) cases were from Kality and Akaki respectively.Ten colonies of rock hyraxes (Heterohyrax brucei) were identified in the vicinities of the 3 localities. Three of the 48 hyraxes (6.3%) trapped from the surroundings harbored natural infections of Leishmania aethiopica. Confirmation of the Leishmania species of the 3 isolates was achieved by PCR amplification and RFLP analysis of the ribosomal DNA internal transcribed spacer (ITS) sequences. Based on sandfly species composition and proximity of resting sites to human settlements, Phlebotomus longipes is circumstantially proven to be the vector of CL in south east Addis Ababa.ConclusionThe study proves the existence of isolated zoonotic foci of CL in south eastern Addis Ababa, with P. longipes as the likely vector and H. brucei as the natural reservoir host.

Field investigation on the repellent activity of some aromatic plants by traditional means against Anopheles arabiensis and An. pharoensis (Diptera: Culicidae) around Koka, central Ethiopia

A study was undertaken to evaluate the impact of traditional application methods of mosquito repellent plants in the reduction of the human-vector contact of malaria vectors in central Ethiopia. The plants (Corymbia citriodora, Eucalyptus camaldulensis, Ocimum suave and Ocimum basilicum) were tested by thermal expulsion and direct burning on traditional stoves in the field against two important malaria vectors in Ethiopia (Anopheles arabiensis and An. pharoensis). A Latin-square design was applied for randomly assigning the treatment plants and control to experimental houses over different nights. The percentage repellency of each candidate plant by both application methods was estimated from the catches of mosquitoes in the treatment and control houses. On direct burning of the plants, O. basilicum showed the highest percentage repellency (73.11%, P<0.001) and E. camaldulensis the least repellency (65.29%, P<0.001) against An. arabiensis. By the same method of application, C. citriodora on the other hand gave the highest repellency (72.87%, P<0.001) while E. camaldulensis was still the least repellent plant (66.60%, P<0.001) against An. pharoensis. On thermal expulsion, C. citriodora exhibited the highest repellency (78.69%, P<0.001) while E. camaldulensis was the lowest repellent plant (71.91%, P<0.001) against An. arabiensis. Against An. pharoensis, C. citriodora gave the highest repellency (72.9%, P<0.001) while E. camaldulensis still gave the least repellency (72.2%, P<0.001) on the same method of application. All the tested plants by both methods of application gave partial but significant protection (>65%) against the house-entry and biting of two important malaria vectors in Ethiopia, and thus have a potential to be used at least as supplements to other control methods. However, feasibility and actual impact on disease transmission need to be known on these and other potentially useful plants.

Impact of housing condition on indoor-biting and indoor-resting Anopheles arabiensis density in a highland area, central Ethiopia

BackgroundExposure of individuals to malaria infection may depend on their housing conditions as houses serve as biting and resting places of vectors. This study describes the association of housing conditions with densities of indoor-biting and indoor-resting Anopheles arabiensis in Hobe, Dirama and Wurib villages of a highland area in central Ethiopia.MethodsData on housing conditions, including presence of house apertures, number of occupants and number and the type of domestic animal tethered inside, were collected. Indoor-biting mosquitoes were sampled using Centers for Disease Control (CDC) light traps and indoor-resting mosquitoes sampled with pyrethrum spray catches (PSCs) monthly for two years (July 2008 to June 2010). Female anophelines were identified to species and processed. Univariate and general linear estimating equation allowing for repeated measures were used to assess the contribution of housing conditions for indoor-biting and indoor-resting An. arabiensis.ResultsAbout 96% (4,597/4,788) of anophelines were caught inside residential houses. Nine anopheline species were identified, among which An. arabiensis was most prevalent (2,489; 52%). Vectors entering houses were higher in those situated at low (β = 4.475; 95% CI = 3.475-5.476; p <0.001; β = strength of the association) and medium (β = 2.850; 95% CI = 1.975-3.724; p <0.001) altitudes compared to high altitude, and where houses have no windows (β = -0.570; 95% CI = -1.047-0.094; p = 0.019) compared with those that have. Numbers of indoor-resting vectors were higher in those situated at low (β = 6.100; 95% CI = 4.571-7.629; p <0.001) and medium (β = 4.411; 95% CI = 2.284-6.537; p <0.001) altitudes compared to high altitudes, and where houses had open eaves (β =1.201; 95% CI = 0.704-1.698; p <0.001) compared with those that had closed eaves.ConclusionHousing conditions such as presence of open eaves, absence of window, location at low and mid altitudes, were strong predictors of indoor exposure to An. arabiensis bite in a highland area of south-central Ethiopia.

Breeding Sites of Aedes aegypti: Potential Dengue Vectors in Dire Dawa, East Ethiopia.

Background and Objectives. Entomological survey was carried out from May-June to September-October 2014 to investigate the presence of dengue vectors in discarded tires and artificial water containers in houses and peridomestic areas. Methods. A cross-sectional immature stage survey was done indoors and outdoors in 301 houses. Mosquito larval sampling was conducted using pipette or dipper depending on container types. Larvae were identified morphologically and larval indices were also calculated. Results. A total of 750 containers were inspected, and of these 405 were positive for mosquito larvae. A total of 1,873 larvae were collected and morphologically identified as Aedes aegypti (n = 1580: 84.4%) and Culex (n = 293: 15.6%). The larval indices, house index, container index, and breteau index, varied from 33.3 to 86.2, from 23.2 to 73.9, and from 56.5 to 188.9, respectively. Conclusion. Aedes aegypti is breeding in a wide range of artificial containers. To control these mosquitoes, the integration of different methods should be taken into consideration.