Araya Gebresilassie

Attraction of phlebotomine sand flies to baited and non-baited horizontal surfaces

Female phlebotomine sand flies (Diptera: Psychodidae) transmit leishmaniasis as they engorge on vertebrate blood required for egg production. Phlebotomus (Phlebotomus) papatasi (Scopoli, 1786), the vector of Leishmania major (Yakimoff & Schokhor, 1914), the causative agent of cutaneous leishmaniasis (CL) were not attracted to large horizontal sticky traps (LHSTs) unless these were baited with CO2 derived from dry ice or from fermenting sugar/yeast mixture (SYM). Attraction of P. papatasi males by CO2 may indicate their tendency to mate on or near the blood-host. Male P. (Larroussius) orientalis (Parrot, 1936), the vector of visceral leishmaniasis (VL) in Ethiopia, were collected on LHSTs in large numbers. Although the number of females remained low, augmentation with SYM, increased the number of females by 800% while the number of males increased by only about 40%. Apparently, male P. orientalis utilize the horizontal surfaces for forming mating swarms. P. (Paraphlebotomus) sergenti (Parrot, 1917), is the vector of CL caused by Leishmania tropica. Although approximately twice as many P. sergenti males were caught on LHSTs as females, it appears that LHSTs were attractive to both sexes. Use of SYM baits is potentially useful for monitoring phlebotomine sand flies in places where dry ice is unobtainable or prohibitively expensive. LHSTs can provide an inexpensive alternative to CDC traps for monitoring some species of sand flies. Unfortunately, the numbers of female sand flies, crucial for estimating transmission of Leishmania, is usually low on LHSTs.

Species composition of phlebotomine sand flies and bionomics of Phlebotomus orientalis (Diptera: Psychodidae) in an endemic focus of visceral leishmaniasis in Tahtay Adiyabo district, Northern Ethiopia

BackgroundVisceral 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.MethodsSand 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.ResultsIn 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.ConclusionsOverall, 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.

Host choice of Phlebotomus orientalis (Diptera: Psychodidae) in animal baited experiments: a field study in Tahtay Adiyabo district, northern Ethiopia

BackgroundHost 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.MethodsTwo 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.ResultsSignificant 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%).ConclusionThe 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.

The influence of moonlight and lunar periodicity on the efficacy of CDC light trap in sampling Phlebotomus (Larroussius) orientalis Parrot, 1936 and other Phlebotomus sandflies (Diptera: Psychodidae) in Ethiopia

BackgroundPhlebotomus 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.MethodsTrapping 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.ResultsIn 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).ConclusionResults 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.

Habitat preference and seasonal dynamics of Phlebotomus orientalis in urban and semi-urban areas of kala-azar endemic district of Kafta Humera, northwest Ethiopia

Visceral leishmaniasis is a significant public health problem in northwest Ethiopia, particularly in Kafta Humera district. The study was designed to determine the species composition and population dynamics of sand flies in five villages representing urban and semi-urban areas of Kafta Humera district namely, Setit Humera, Mykadra, Rawyan, Bereket and Adebay. Sand flies were collected for two to three nights monthly from May 2011 to April 2012 using CDC light-traps and sticky traps. Traps were placed in villages, at periphery of villages and farm fields. Sticky traps were also used for sampling indoor active sand flies. In total, 13,097 sand fly specimens of eight Phlebotomus species and 91,949 Sergentomyia specimens were collected. Among the Phlebotomus, P. orientalis was the predominant species (58.1%) followed by P. papatasi (29.6%), P. lesleyae (5.6%), P. bergeroti (3.8%), P. duboscqi (2.1%), P. alexandri (0.4%), P. heischi (0.2%) and P. rodhaini (0.2%). Significantly, higher number of P. orientalis was caught in Adebay village and the least in Setit Humera town. Seasonal abundance of P. orientalis increased during the dry season (January-May) and dropped drastically in the wet season (late June-September). Significant positive correlation was found between monthly abundance of P. orientalis and the monthly average air and surface soil temperature, while a negative correlation was found with monthly average rainfall and relative humidity. Higher number of P. orientalis was collected outdoors, especially in the farm fields followed by periphery of villages. Thus, various observations strongly suggested P. orientalis to be the principal vector in the study areas, where farm lands and periphery of villages were identified as the most risky habitats, whereas the indoors were the least ones. Appropriate control methods should be designed and implemented according to the knowledge of P. orientalis habitat preferences and seasonal dynamics in the district.

Studies on sand fly fauna and ecological analysis of Phlebotomus orientalis in the highland and lowland foci of kala-azar in northwestern Ethiopia

Background 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. Methods 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. Results 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 (P<0.05). However, mean density of P. orientalis in the two areas in sticky trap collections had positive and insignificant association with the temperature (P>0.05). 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). Conclusions 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.

Diversity and altitudinal distribution of phlebotomine sand flies (Diptera: Psychodidae) in visceral leishmaniasis endemic areas of northwest Ethiopia

Background The Leishmaniases are caused by the protozoan parasites of the genus Leishmania and are transmitted to humans by the bite of infected female phlebotomine sand flies. Both visceral and cutaneous leishmaniases are widely distributed in different parts of Ethiopia. The aim of this study was to determine the diversity and altitudinal distribution of phlebotomine sand flies from Kafta Humera to Gondar town in northwest Ethiopia. Methods Seven localities were selected with distinct altitudinal variations between 550 m above sea level (m a.s.l) and 2300 m a.s.l. In each locality, sand flies were collected using standard CDC light traps and sticky traps during the active sand fly season from December 2012 to May 2013. Shannon-Weiner species diversity index and Jaccard’s coefficient were used to estimate species diversity and similarity between altitudes and localities, respectively. Results A total of 89,044 sand flies (41,798 males and 47, 246 females) were collected from the seven localities/towns throughout the study period. Twenty-two species belonging to 11 species in the genus Phlebotomus and 11 species in the genus Sergentomyia were documented. Of these, Sergentomyia clydei (25.87%), S. schwetzi (25.21%), S. africana (24.65%), S. bedfordi (8.89%), Phlebotomus orientalis (6.43%), and S. antennata (4.8%) were the most prevalent species. The remaining 10 Phlebotomus species and six Sergentomyia were less frequent catches. In CDC light trap and sticky trap, higher species diversity and richness for both male and female sand flies was observed at low altitude ranging from 550 to 699 m a.s.l in Adebay village in Kafta Humera district whereas low species richness and high evenness of both sexes were also observed in an altitude 1950–2300 m a.s.l. Conclusion The results revealed that the presence of leishmaniasis vectors such as P. orientalis, P. longipes, P. papatasi, and P. duboscqi in different altitudes in northwest Ethiopia. P. orientalis a vector of L. donovani, occurred between altitude 500–1100 m a.s.l, the area could be at high risk of VL. P. longipes a vector of L. aethiopica, was recorded in the highland area in Tikil-Dingay and Gondar town, implicating the possibility of CL transmission. Hence, further investigation into vector competence in relation to leishmaniasis (VL and CL) in the region is very vital.

Physiological Age Structure and Leishmania spp. Detection in Phlebotomus (Larroussius) orientalis (Parrot, 1936) (Diptera: Psychodidae) at an Endemic Focus of Visceral Leishmaniasis in Northern Ethiopia

Visceral leishmaniasis (VL) caused by Leishmania donovani is endemic in northern Ethiopia, where P. orientalis is the most important presumed vector. This study was designed to determine the physiological age structure and the occurrence of Leishmania infection in the vector of VL in Tahtay Adiyabo district, northern Ethiopia. Sand flies were collected using CDC light traps from peridomestic and agricultural fields between May 2011 and April 2012 and P. orientalis females were dissected for age determination and detection of Leishmania promastigotes. Sand flies were also analyzed for L. donovani detection using molecular methods. Of 1,282 P. orientalis examined for abdominal stages and age characterization, 66.2%, 28.2%, 4.1%, and 1.6% were unfed, freshly fed, half-gravid, and gravid. Parous rate in unfed females was 34.1% and 35.4% in peridomestic and agricultural fields, respectively. Out of 921 P. orientalis females dissected, one specimen (0.1%) was found naturally infected with promastigotes. Five pools (25 females) of unfed P. orientalis were also found with DNA of Leishmania spp. In particular, a single P. orientalis was positive for L. donovani (0.5%). Based on this and other evidences (abundance, human blood feeding, and xenodiagnostic studies), P. orientalis is the principal vector of VL in this endemic focus.

Some aspects of entomological determinants of Phlebotomus orientalis in highland and lowland visceral leishmaniasis foci in northwestern Ethiopia

Visceral leishmaniasis (VL) is one of the major public health problems in northwest Ethiopia, mainly in Libo-Kemkem and Metema districts, where Phlebotomus orientalis is the most probable vector of the disease. The aim of this study was to determine the physiological age, host preference and vectorial potential of P. orientalis in the highland and lowland foci of the region. Sand flies were collected using CDC light traps between May 2011 and April 2012 in Libo-Kemkem and October 2012 and September 2013 in Metema from household compounds, farm field and mixed forest. Females belonging to Phlebotomus were dissected for physiological age determination and Leishmania detection and isolation. Leishmania infections in sand flies were investigated using molecular methods. Freshly fed Phlebotomus females were tested to identify blood meal sources using PCR-RLB and ELISA. A total of 1149 (936 from Libo-Kemkem and 213 from Metema) blood unfed female P. orientalis were dissected for age determination. The parity rate was 45.6% and 66.2% in Libo-Kemkem and Metema, respectively. None of 798 female P. orientalis dissected (578 from Libo-Kemkem and 220 from Metema) was infected with Leishmania parasites. A total of 347 P. orientalis specimens collected from Libo-Kemkem were processed using PCR, of which 10 (2.8%) specimens were found with DNA of Leishmania spp. Of a total 491 freshly fed female P. orientalis analyzed for blood meal origins by RLB-PCR and ELISA, 57.6% (67.8% from Libo-Kemkem and 49.8% from Metema) were found to contain bovine blood while 4.9% (3.7% from Libo-Kemkem and 5.7% from Metema) were of human blood. In conclusion, the present study showed parity difference between the two populations of P. orientalis and that both populations have strong zoophilic behavior. Based on the presented evidences, the species is strongly implicated as a vector of kala-azar in both areas. Therefore, vector control should be a component of a strategy to manage visceral leishmaniasis in both study areas.

First detection of Anopheles stephensi Liston, 1901 (Diptera: culicidae) in Ethiopia using molecular and morphological approaches

Malaria is a major public health concern in Ethiopia. With the increase in malaria cases in the Somali Region of Ethiopia, understanding the distribution and identifying the species of malaria vectors is vital to public health. Here we report the first detection of Anopheles stephensi in Ethiopia, a malaria vector typically found in the Middle East, the Indian subcontinent, and China, but recently found in Djibouti. An entomological investigation was conducted during November to December 2016 in Kebri Dehar town of the Ethiopian Somali Regional State as ancillary work for Anopheles spp. surveillance. Mosquito larvae were collected from water reservoirs. Larvae were reared in the laboratory to the adult stage and identified morphologically. PCR and sequencing of cytochrome oxidase 1 (COI) and internal transcribed spacer 2 (ITS2) loci were performed. Basic Local Alignment Search Tool (BLAST) was used to compare sample sequences to sequences in the NCBI nucleotide database for species identification. To further analyze the relationship between the specimen we collected in Kebri Dehar and other Anopheles samples available in Genbank, phylogenetic analysis was performed using a maximum likelihood approach. Molecular and morphological results confirm specimens were An. stephensi. The closest high scoring hit was for all specimens was for the An. stephensi sequence. Independent phylogenetic analyses of COI and ITS2 sequences revealed in both cases strong bootstrap (100) support for our sequence forming a clade with other An. stephensi sequences to the exclusion of any other species of Anopheles. In conclusion, Anopheles stephensi is present in Kebri Dehar town in Ethiopia. These findings highlight the need for additional research to examine the role of An. stephensi in malaria transmission in Ethiopia.