Gebresilassie et al. Parasites & Vectors (2015) 8:190 DOI 10.1186/s13071-015-0807-4 RESEARCH Open Access Host choice of Phlebotomus orientalis (Diptera: Psychodidae) in animal baited experiments: a field study in Tahtay Adiyabo district, northern Ethiopia Araya Gebresilassie1,2*, Solomon Yared2,4, Essayas Aklilu1, Oscar David Kirstein3, Aviad Moncaz3, Habte Tekie1, Meshesha Balkew4, Alon Warburg3, Asrat Hailu5 and Teshome Gebre-Michael4 Abstract Background: 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. Methods: 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 attractedsandflieswerecollectedusingunlitCDCtrapsfor10 trapping nights. Sand fly specimens collected from each of the experiments were identified to species level and counted. Results: 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%). Conclusion: 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. Keywords: Host attractiveness, Phlebotomus orientalis, Visceral leishmaniasis, Zoophilic sand flies * Correspondence: [email protected] 1Department of Zoological Sciences, Addis Ababa University, Addis Ababa, Ethiopia 2Department of Biology, College of Natural Science, Jigjiga University, Jigjiga, Ethiopia Full list of author information is available at the end of the article © 2015 Gebresilassie et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Gebresilassie et al. Parasites & Vectors (2015) 8:190 Page 2 of 10 Background Methods Ninety-eight countries and 3 territories on 5 continents Study area are endemic for either of the two major forms of leish- The study area has previously been described elsewhere maniasis: cutaneous leishmaniasis (CL), a disfiguring and [22]. Briefly, the investigation was conducted in the Geza stigmatizing disease, and visceral leishmaniasis (VL) or Adura sub-village in the Tahtay Adiyabo district (14°22’27” kala-azar, which is fatal if untreated [1,2]. In Ethiopia, N/37°44’36” E) in Tigray Regional State, Northern VL caused by Leishmania donovani, is considered an Ethiopia, which is situated 1,117 km north of Addis emerging disease with an estimated incidence of 3,700 to Ababa. The climate is generally sub-tropical-arid, with an 7,400 cases per year [2]. Highly VL endemic foci are in the extended dry period of nine to ten months (October- south‐west and the Humera and Metema lowlands in the May), experiencing only one rainy season (June-September) north‐west of the country [3,4]. However, recently increas- with a mean annual precipitation of about 600 mm ing numbers of VL has been reported from previously (Ethiopian National Meteorological Agency). April and non-endemic regions such as Libo Kemkem district of May are the warmest months with an average temperature Amhara Regional State and Tahtay Adiyabo district in of 39°C and January is the coldest with an average Tigray Regional State in Northern Ethiopia [5,6]. temperature of 14.2°C. The various forms of leishmaniasis, including VL are In different villages of Tahtay Adiyabo district, large transmitted by the bite of infected female sand flies of numbers of domestic animals including cattle, sheep, the genus Phlebotomus in the Old World and Lutzomyia goats, donkeys, camels, dogs and chickens are kept. More- in the New World [7,8]. Transmission of VL occurs over, a wide range of wild animals such as hares, ground when a sand fly acquires infection during feeding on an squirrels, rodents, reptiles, white-tailed mongoose and infected host and transmits the parasite during subse- foxes are either occasionally or commonly seen. quent feeding after completion of the gonotrophic cycle, during which the parasite multiplies in the midgut and mi- Host choice experiments grates to the foregut and mouthparts of the infected sand The two independent host choice studies under field fly female. In southern Ethiopia and Kenya, the principal conditions were conducted between March and April vector is P. martini which breeds and rests in termite 2013 in rotational experimental design in which wild mounds [9,10], whereas in northern Ethiopia and eastern sand flies were given the choice of different animal baits. Sudan, P. orientalis is implicated as the vector inhabiting The experiments were conducted in an open agricultural Acacia forests and cracking vertisols [11-13]. field, where there were no potential bloodmeal source The host preferences of several sand fly species have animals (cattle, sheep, goats, donkeys, camels, dogs, been investigated mostly through the identification of the chickens and other small wild animals) near the test sources of bloodmeals using serological [14,15] or molecu- traps for at least 400–450 meters. lar assays [16,17] and host attractiveness experiments [18-20]. Previously, the host preference of P. orientalis in Experiment I Sudan and Ethiopia was determined by quantifying the The experiment was conducted using wild sand flies of- host preferences using different animal baits [20,21] or by fered a choice of seven baits (human volunteer, cow, identification of sources of bloodmeals by ELISA [12]. In sheep, goat, donkey, dog and chicken) and control (with- eastern Sudan, it has been shown that P. orientalis is out baits) assigned in tent traps. Each tent trap (dimen- largely attracted to dogs, which is a suspected reservoir sion: 2.5 m × 2.5 m × 2.5 m) was constructed from host for domestic transmission of VL in the area [13]. In transparent sand fly-proof netting supported by four the Humera-Metema plains of northwest Ethiopia, the rectangular metal frames and four metal poles to firmly host preference of P. orientalis appeared to be zoophilic, fix them to the ground when installed (Figure 1-A). Six predominantly feeding on bovine blood [13]. These lim- cone-shaped openings were fitted on the sides of the tent ited studies in East Africa might indicate P. orientalis to and the tent was raised a few centimeters above the ground be an opportunistic feeder; however, detailed studies are to allow entry of host seeking sand flies (Figure 1-B). One needed to understand the natural host preference profile side of the net had a long zipped slit (top to bottom) to en- of P. orientalis and other sand fly species and their pos- able entry and exit of host plus the participant to aspirate sible epidemiological significance of both domestic and sand flies from inside the trap. The tent trap design was wild animals in the transmission dynamics of VL. based on a prototype used previously to assess sand fly at- Taking into account the fragmentary information avail- traction behavior in Colombia [19]. able on the feeding habit of P. orientalis,wedesignedan The animals were tethered in the center of the tent experiment to determine the relative host attractiveness while the human volunteer slept on a cot protected by and feeding success of P. orientalis on domestic and small an untreated sand fly-proof mosquito net inside the tent wild mammals in a VL endemic area of north Ethiopia. trap. Baited tent trap and an un-baited control trap were Gebresilassie et al. Parasites & Vectors (2015) 8:190 Page 3 of 10 Figure 1 Host attractiveness experiments. A: Tent trap with animal bait. B: Cones fitted to one side of the tent. C: Arrangement of animal baits in the field. D: Cage trap baited with ground squirrel. E: Cage baited with rodent species. arranged in a circular manner at a distance of about 30 cahirinus). Hares (Lepus sp.) were captured by chasing meters from each other (Figure 1-C). The baits and un- them from their hidings in the bush. baited traps rotated every night between different posi- Square box traps (30cmx30cmx30cm) with entrance tions to eliminate location or site variations in sand fly brass screen cones on the three sides were locally con- abundance. Two replicate collections were performed on structed based on the design of Turner and Hoogstraal a total of 16 trapping nights.