Kirstein et al. Parasites & Vectors 2013, 6:341 http://www.parasitesandvectors.com/content/6/1/341 RESEARCH Open Access Attraction of Ethiopian phlebotomine sand flies (Diptera: Psychodidae) to light and sugar-yeast mixtures (CO2) Oscar D Kirstein1, Roy Faiman1,4, Araya Gebreselassie2, Asrat Hailu3, Teshome Gebre-Michael2 and Alon Warburg1* Abstract Background: Visceral leishmaniasis (VL) known as Kala-Azar is a serious systemic disease caused by Leishmania donovani parasites (Trypanosomatidae: Kinetoplastida). The disease is prevalent in the Indian Sub-continent, East Africa and Brazil. In Africa, the worst affected regions are in Sudan, with an estimated 15,000-20,000 cases annually and Ethiopia with 5,000-7,000 cases a year. The main vector of VL in Sudan and Northern Ethiopia is Phlebotomus orientalis, a sand fly frequently found in association with Acacia spp and Balanite spp woodlands. Methods: To optimize sampling of sand flies for epidemiological studies in remote areas we tested different means of attraction. Miniature suction traps employing 2AA batteries (3 V) were deployed in the up-draft orientation and baited with chemical light-sticks (Red, Yellow and Green), or bakers’ yeast in sugar solution (emitting CO2 and perhaps other attractants). These traps were compared with standard CDC incandescent light traps employing 6 V batteries. Trials were conducted during two consecutive years at different localities around Sheraro, a town in West Tigray, Northern Ethiopia. Results: The sand fly species composition was similar but not identical in the different locations tested with different Sergentomyia spp. predominating. Phlebotomus spp. comprised less than 1% of the catches during the first year trials (November – December 2011) but increased markedly during the second year trials performed later in the dry season at the height of the sand fly season in February 2012. Although there did not appear to be a species bias towards different light wave-lengths, fermenting yeast in sugar solution attracted relatively more Phlebotomus spp. and Sergentomyia schwetzi. Conclusions: Although the standard 6 V CDC incandescent light traps captured more sand flies, light-weight (~350 g) 3 V suction traps baited with chemical light-sticks were shown to be effective means of monitoring sand flies. Such traps operated without light and baited with yeast in sugar solution caught relatively more Phlebotomus spp. Keywords: Attractants for blood-sucking insects, Chemical light-stick, Ethiopia, Phlebotomus orientalis, Sergentomyia, Visceral leishmaniasis, Yeast fermentation * Correspondence: [email protected] 1Department of Microbiology and Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious and Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, Jerusalem 91120, Israel Full list of author information is available at the end of the article © 2013 Kirstein et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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. Kirstein et al. Parasites & Vectors 2013, 6:341 Page 2 of 9 http://www.parasitesandvectors.com/content/6/1/341 Background sucking insects and are considered superior to CDC light Phlebotomine sand flies (Diptera: Psychodidae) are of traps for both mosquitoes and sand flies. Moreover, CO2- considerable public health importance in many regions baited traps have the added advantage that most moths of the world where they transmit the causative agents of and beetles are not attracted by CO2 so the yield is usually leishmaniasis, bartonellosis and sand fly fever [1-5]. The considerably “cleaner” [29-32]. CO2 for baiting traps is leishmaniases are a group of diseases caused by parasites routinely supplied from either pressurized cylinders fitted of the genus Leishmania, affecting mainly the poorest of with pressure valves or from dry ice [29,30]. Such traps the poor in developing countries; 350 million people are typically emit CO2 at concentrations that are roughly two considered at risk of contracting leishmaniasis, and some orders of magnitude higher than background levels two million new cases occur yearly [6,7]. The proven (20,000 vs. 400 ppm) [33]. Blood seeking female mosqui- vectors of leishmaniasis in the old world belong to the toes and sand flies are generally thought to respond to genus Phlebotomus Rondani & Berté, 1840 and new small changes in CO2 concentration by flying upwind and world vectors to the genus Lutzomyia Franca & Parrot, maneuvering between concentration gradients and visual 1920 [1,8,9]. cues until reaching their target [30,34]. Unfortunately, nei- Visceral leishmaniasis (VL) or Kala-Azar, caused by ther dry ice nor pressurized gas cylinders are easily found infection with Leishmania donovani complex parasites is in remote areas of developing countries where many leish- prevalent in 65 countries. The majority (90%) of cases maniasis foci are found. Sugar solutions fermented by occur in poor rural and suburban areas of 5 countries: Bakers’ yeast (sugar/yeast mixtures [SYM]) were demon- Bangladesh, India, Nepal, Sudan and Brazil [7,10]. In strated to be attractive to Anopheline mosquitoes in East Africa VL is widely distributed throughout much of Tanzania [35]. We have previously demonstrated the Sudan, the lowland regions of Ethiopia as well as Kenya efficacy of both SYM and chemiluminescent light-sticks as and South Sudan [7,11,12]. In Ethiopia, important en- attractants for phlebotomine sand flies in Ethiopia and in demic foci include the Humera and Metema plains in Israel using horizontal sticky traps [36]. Here, we com- northwestern Tigray and the Segen and Woyto river bined these attractants with inverted up-draft suction valleys in the southwest [11,13]. traps similar to CDC traps but fabricated in our laboratory The sand fly Phlebotomus (Larroussius) orientalis and designed to operate on 2 AA type rechargeable batter- Parrot, 1936 is the main vector of VL in Sudan and in ies (3 V). Such traps weigh less than 350 g each making it Northern Ethiopia where it is frequently associated with possible to carry and deploy a large number in areas Acacia seyal and Balanites aegyptiaca woodlands grow- where vehicle access is restricted. These 3 V traps are not ing in black cotton soils [14-16]. P. orientalis is primarily equipped with incandescent lights because 2 AA batteries a lowland species, but in Ethiopia it is also found in do not provide sufficient power to run both motor and altitudes of up to 2,000 m [17]. In the more southerly light for 12 hours. Therefore, to experiment with light VL foci, P. orientalis, P. martini and P. celiae have been traps we utilized chemiluminescent light-sticks to attract implicated as vectors of L. donovani [18-20]. the sand flies to the 3 V traps [36]. Sand flies belonging to the genus Sergentomyia are the The current study forms part of a comprehensive predominant phlebotomine species in many African project on the ecology and transmission dynamics of ecosystems [21-23]. Sergentomyia spp. are known to visceral leishmaniasis in Ethiopia. The experiments transmit Leishmania that infect lizards (previously Saur- reported here were conceived and designed to optimize oleishmania) [24]. Although most species are preferen- and facilitate large scale trapping of sand flies using tial reptile-feeders certain species also bite mammals equipment and materials appropriate for remote areas. including humans and were suspected of transmitting At the same time, the results comprise the only entomo- human leishmaniasis [25,26]. In our study area, Sergento- logical data pertaining to this important focus of kala myia spp. were exceptionally abundant and some species azar in the Sheraro district of northern Ethiopia. were found to feed on humans (Moncaz et al. unpub- lished). This finding, coupled with the almost total Methods absence of Phlebotomus spp. at certain times of the year, Experimental sites warranted the inclusion of Sergentomyia spp. in the The study was carried out during the dry seasons of current study despite their dubious role as vectors of 2011–12 in rural environments near the town of Sheraro leishmaniasis to humans [25,27]. (14° 24′ 09.69″N – 37° 46′ 39.69″E). Sheraro town is a Centers for disease control (CDC) miniature light sub-district of the Tahtay Adiabo district, located in the traps that are frequently used for collecting sand flies Semien Mi’irabawi Zone (Northwestern) of Tigray Region, attract sand flies from short distances not much exceeding northern Ethiopia at an elevation of circa 1,000 m above two meters [28]. CDC traps baited with carbon dioxide sea level. The climate is semi-arid typical of the African (CO2) have been used extensively for trapping blood- Sahel, with an extended dry period of nine to ten months Kirstein et al. Parasites & Vectors 2013, 6:341 Page 3 of 9 http://www.parasitesandvectors.com/content/6/1/341 with most all of the rains (~600 mm) concentrated in 50 A woodland dominated by Balanites aegyptiaca trees near to 60 days during June-September. The mean annual the village of Erdweyane (Figure