Blood-feeding behavior studies are important for estimating the efficiency of pathogen transmission and assessing the relative human disease risk. However, in Ethiopia and other parts of East Africa there are large remaining gaps in identifying the feeding habits of Phlebotomus orientalis, the vector of Leishmania donovani. The aim of the study was to determine the blood feeding patterns of P. orientalis in Tahtay Adiyabo district, northern Ethiopia. For bloodmeal analysis, sandflies were collected from three different villages of Tahtay Adiyabo district using CDC light traps, sticky traps, and pyrethrum spray catches. Bloodmeal of engorged female sandflies was identified using cytochrome (cyt) b-PCR and reverse-line blotting (RLB) and enzyme linked immunosorbent assay (ELISA) assays. Most (637/641) of the females analyzed were P. orientalis. Successful identification of the host from bloodmeals was achieved in 83.03 and 92.1 % using cyt b PCR-RLB and ELISA, respectively. Bloodmeal analysis of P. orientalis females revealed that they have a range of hosts with predominant preference to bovines followed by donkey, human, goat, sheep, dog, and camel. Results obtained from bloodmeal analyses demonstrate that the feeding preference of P. orientalis is mainly zoophilic, which could vary depending on the availability of hosts.

Host choice and feeding success of sand fly vectors of visceral leishmaniasis (VL) are important factors in understanding the epidemiology and for developing efficient control strategies. The aim of the present study was to determine the host preference of Phlebotomus orientalis in the VL focus of Tahtay Adiyabo district, northern Ethiopia. Two separate experiments were conducted testing attraction of P. orientalis to humans, domestic animals, and small wild animals. The host choice of P. orientalis and other sand fly species was assessed using tent traps baited with seven different animals (human, cow, sheep, goat, donkey, dog and chicken) and a blank control. Baited traps were rotated every night in a Latin square design for two consecutive full rounds totaling 16 trap-nights. The second set of experiments tested attraction to small wild animals including; ground squirrel (Xerus rutilus), hare (Lepus sp.), gerbil (Tatera robusta) and spiny mouse (Acomys cahirinus). Animals were caged in standard rodent traps or cylindrical wire-mesh cages. The bait animals were placed in agricultural field and the attracted sand flies were collected using unlit CDC traps for 10 trapping nights. Sand fly specimens collected from each of the experiments were identified to species level and counted. Significant difference (P < 0.05) was observed in the attraction and feeding rate of P. orientalis to different baits. In the first experiment, cow-baited tent traps attracted the highest mean number of P. orientalis (mean = 510 flies). The engorgement rate of P. orientalis on donkey was the highest followed by cow, and much lower on goat, sheep, dog and chicken. In the case of smaller wild animals, more numbers of P. orientalis females were attracted to squirrels followed by hares, gerbils and the spiny rat. However, the engorgement rates for P. orientalis in the smaller animals were very low (1.08%) compared with larger domestic animals (30.53%). The tendency of female P. orientalis to engorge in large numbers on certain species of domestic as well as wild animals strongly indicated that the species is primarily zoophilic in its host preference with feeding habits that may vary depending on the availability of hosts.

Visceral leishmaniasis (VL) is a neglected tropical disease, which is strongly associated with poverty. VL caused by Leishmania donovani and transmitted by Phlebotomus orientalis is endemic in various remote areas of north and north-west Ethiopia. The present study was designed to determine the sand fly fauna and bionomics of P. orientalis in the VL endemic focus of Tahtay Adiyabo district. Sand flies were collected using CDC light traps (n = 602), sticky traps (n = 9,350) and indoor pyrethrum spray catches (n = 578 house visits) from indoor, peri-domestic and agricultural field habitats between May 2011 to April 2012. All sand fly specimens collected were identified to species level and counted. In total, 100,772 sand fly specimens, belonging to 25 sand fly species (nine Phlebotomus and sixteen Sergentomyia) were collected and identified. S. africana and P. orientalis made up 59.1% and 23.5% of the collected sand flies, respectively. As it could be determined from the proportion of collections from outdoor (peri-domestic and agricultural fields) and indoor locations, P. orientalis appears to exhibit increased exophilic behavior. The outdoor to indoor index was 79:1 on m2 of sticky traps. Mean density of P. orientalis caught was significantly higher on horizontally placed sticky traps (mean = 60 ± 14.56/m2/night) than vertically deployed sticky traps (12 ± 3.57/m2/night). The highest abundance of P. orientalis occurred between March and April. Through July to September, there was a sharp decline in abundance of P. orientalis population. Regarding climatic variables, P. orientalis density in light traps and on sticky traps showed a significant positive and negative association with temperature and relative humidity, respectively. However, non-significant negative correlation was observed with rainfall pattern. Overall, P. orientalis was found to be the most abundant Phlebotomus species, showing marked seasonal abundance that mainly peaks during the dry season (March to April). Likewise, the people in the area usually sleep in compounds during these months that potentially expose them to a high risk of peri-domestic VL transmission.

Phlebotomus orientalis is the main sandfly vector of visceral leishmaniasis in the north and northwest of Ethiopia. CDC light traps and sticky traps are commonly used for monitoring sandfly populations. However, their trapping efficiency is greatly influenced by various environmental factors including moonlight and lunar periodicity. In view of that, the current study assessed the effect of moonlight and lunar periodicity on the performance of light traps in collecting P. orientalis. Trapping of P. orientalis and other Phlebotomus spp. was conducted for 7 months between December 2012 and June 2013 using CDC light traps and sticky traps from peri-domestic and agricultural fields. Throughout the trapping periods, collections of sandfly specimens were carried out for 4 nights per month, totaling 28 trapping nights that coincided with the four lunar phases (viz., first quarter, third quarter, new and full moon) distributed in each month. In total, 13,533 sandflies of eight Phlebotomus species (P. orientalis, P. bergeroti, P. rodhaini, P. duboscqi, P. papatasi, P. martini, P. lesleyae and P. heischi) were recorded. The predominant species was P. orientalis in both trapping sites and by both methods of collection in all lunar phases. A significant difference (P < 0.05) was observed in the mean numbers of P. orientalis and other Phlebotomus spp. caught by CDC light traps among the four lunar phases. The highest mean number (231.13 ± 36.27 flies/trap/night) of P. orientalis was collected during the new moon phases, when the moonlight is absent. Fewer sandflies were attracted to light traps during a full moon. However, the number of P. orientalis and the other Phlebotomus spp. from sticky traps did not differ in their density among the four lunar phases (P = 0.122). Results of the current study demonstrated that the attraction and trapping efficiency of CDC light traps is largely influenced by the presence moonlight, especially during a full moon. Therefore, sampling of sandflies using light traps to estimate population density and other epidemiological studies in the field should take the effect of moonlight and lunar periodicity into account on the trapping efficacy of light traps.

Visceral leishmaniasis (VL) also known as kala-azar is a growing health problem in Ethiopia with an estimated annual VL incidence between 3700 and 7400. The disease is mainly endemic in northwestern parts of the country. The aim of the current study was to determine the sand fly fauna and ecology of Phlebotomus orientalis in two endemic and ecologically distinct areas of northwestern Ethiopia. Sand flies were collected using CDC light traps, sticky traps and pyrethrum spray catches from peri-domestic, mixed forest, farm field and indoor habitats from both Libo-Kemkem (May 2011-April 2012) and Metema (October 2012-September 2013) districts. A total of 51,411 sand fly specimens were collected and identified (10,776 from highland and 40, 635 from the lowland areas). Seven species were found in the highland area: two Phlebotomus spp. (P. orientalis and P. rodhaini) and five Sergentomyia species. Whereas 19 species were found in the lowland area: six Phlebotomus (P. orientalis, P. rodhaini, P. bergeroti, P. duboscqi, P. papatasi and P. martini) and 13 Sergentomyia species. Of the Phlebotomus spp., P. orientalis was the predominant species in both the highland (99.9%) and lowland (93.7%) areas. Indoor collections using pyrethrum spray catches and sticky traps indicated that P. orientalis has a strong exophilic and exophagic behaviors in both districts. In both areas, this species showed seasonal occurrence and showing abundance during the dry months (March-May/June) of the year and increasing in numbers till the rain season, when numbers dropped dramatically. Mean density of P. orientalis in the two areas had positive and significant correlation with mean temperature in light trap collections (P0.05). Regarding the rainfall pattern, density of P. orientalis had negative and statistically insignificant correlation (for light trap collections for both areas) and significant correlation (for sticky trap collections for lowland area). The current study indicated the variation in sand fly fauna between the highland and lowland districts, wherein, P. orientalis was found to be the most abundant Phlebotomus species. The study also determined that P. orientalis exhibits distinct seasonality, where its abundance increases during the dry season and disappears when the rainy period starts in both study areas. This entomological observation on the bionomics of P. orientalis provides significant evidence for considering vector control or preventive measures in the areas studied.

Malaria is a public health problem in Ethiopia and its transmission is generally unstable and seasonal. For the selection of the most appropriate vector control measures, knowledge on the ecology of the vector is necessary at a local level. Therefore, the objectives of this study were to document the species composition, breeding habitat characteristics and occurrence of anopheline larva in Sheni stream and the vectorial role of the prevailing Anopheles in relation to malaria transmission in Addis Zemen, Ethiopia. Immature anophelines were sampled from breeding habitats and characteristics, such as water temperature, turbidity, water current, water pH and other variables, of the habitats were measured from October 2011 to February 2012. Adult anophelines were sampled inside human dwellings using space spray and Center for Disease Control light traps. Artificial pit shelters and clay pots were also used for outdoor adult collections. Anophelines collected were identified using morphological key. The enzyme-linked immunosorbent assay was applied to detect circumsporozoite proteins of Plasmodium and source of blood meals. A total of 6258 Anopheles larvae were collected and identified morphologically. Five anopheline species were found: An. gambiae (s.l.), An. cinereus, An. demeilloni, An. christi and An. pretoriensis. Anopheles gambiae (s.l.) existed in most of the habitats investigated. Only the former three species were captured in the adult collections. Sun-lit Sheni stream, rain pools, hoof prints, drainage and irrigation canals were found to be habitats of larvae. Anopheles gambiae (s.l.) larvae were most abundantly sampled from sand mining and natural sand pools of Sheni stream. Multiple regression analysis showed that clear, permanent and temporary habitats devoid of mats of algae were the best predictors of An. gambiae (s.l.) larval abundance. It is also the responsible malaria vector in the study area and exhibits anthropophilic and endophagic behaviour. The malaria vector An. gambiae (s.l.) was found in Addis Zemen throughout the study period from both adult and larval collections. Sheni stream is the main larval habitat responsible for the occurrence of anopheline larvae during the dry season of the study area when other breeding sites perish.