Aviad Moncaz

Breeding Sites of Phlebotomus sergenti, the Sand Fly Vector of Cutaneous Leishmaniasis in the Judean Desert

Phlebotomine sand flies transmit Leishmania, phlebo-viruses and Bartonella to humans. A prominent gap in our knowledge of sand fly biology remains the ecology of their immature stages. Sand flies, unlike mosquitoes do not breed in water and only small numbers of larvae have been recovered from diverse habitats that provide stable temperatures, high humidity and decaying organic matter. We describe studies designed to identify and characterize sand fly breeding habitats in a Judean Desert focus of cutaneous leishmaniasis. To detect breeding habitats we constructed emergence traps comprising sand fly-proof netting covering defined areas or cave openings. Large size horizontal sticky traps within the confined spaces were used to trap the sand flies. Newly eclosed male sand flies were identified based on their un-rotated genitalia. Cumulative results show that Phlebotomus sergenti the vector of Leishmania tropica rests and breeds inside caves that are also home to rock hyraxes (the reservoir hosts of L. tropica) and several rodent species. Emerging sand flies were also trapped outside covered caves, probably arriving from other caves or from smaller, concealed cracks in the rocky ledges close by. Man-made support walls constructed with large boulders were also identified as breeding habitats for Ph. sergenti albeit less important than caves. Soil samples obtained from caves and burrows were rich in organic matter and salt content. In this study we developed and put into practice a generalized experimental scheme for identifying sand fly breeding habitats and for assessing the quantities of flies that emerge from them. An improved understanding of sand fly larval ecology should facilitate the implementation of effective control strategies of sand fly vectors of Leishmania.

Characterization of breeding sites of Phlebotomus orientalis – The vector of visceral leishmaniasis in northwestern Ethiopia

We studied breeding sites of Phlebotomus orientalis (Diptera: Psychodidae) the vector of visceral leishmaniasis in northern Ethiopia. Although numbers were rather small, 165 sand flies were captured emerging from vertisol cracks. The most productive breeding sites were cracked vertisols, dry river banks and close to trees. No sand flies were caught emerging from sandy clay loam soils in peri-domestic habitats but a few were captured emerging from gaps in a stone wall. Abiotic parameters in vertisols close to trees and in open field from which P. orientalis had emerged, were compared. Soil pH was slightly alkaline and salinity was low. Organic matter contents were similar in both sites. Temperatures and RH remained relatively stable near trees from the end of the rainy season through mid dry season, yet fluctuated markedly at the shallower depth in the open field. During the rainy season, cracks in the soil were sealed resulting in significant lowering of the oxygen concentrations near the tree. Gravimetric water content of soil near trees was lower than open field at shallow depth but similar deeper down. We conclude that ambient conditions suitable for sand fly larvae at shallow depths (45cm) are restricted to areas close to trees. However, deeper in vertisols (90cm) suitable conditions are apparently maintained throughout the dry season even in open fallow fields.

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.

Sergentomyia spp.: Breeding sites in vertisols and peri-domestic habitats in North West Ethiopia

Sand flies belonging to the genus Sergentomyia Franca & Parrot, 1920, are hematophagous insects feeding mostly on reptiles and birds, but some species feed also on mammals including humans. Sergentomyia spp. frequently comprise the vast majority of sand flies trapped along with Phlebotomus spp., the vectors of mammalian leishmaniasis. Within the framework of a project on the ecology and transmission of visceral leishmaniasis in Ethiopia, putative breeding sites of phlebotomine sand flies were studied. Large horizontal sticky traps (LHSTs) covered with sand fly-proof mesh were deployed over cracked vertisol and related habitats for up to 3 nights, and emerging sand flies were collected daily. Emergence traps (ETs) were also adapted to sample other putative breeding sites including tree trunks, termite mounds, rock piles and vertical river banks. Productive breeding sites were identified in the trunks and roots systems of trees, vertisol fields, cracks and burrows in vertisol dry river banks and termite mounds. Emerging flies were also collected form a stone wall and a rock pile situated inside a village. Significantly more Sergentomyia spp. were trapped in vertisols by ETs deployed over root system than in open fields. Similarly, more sand flies emerged from cracks in the vertisol in fallow Sorghum than in fallow sesame fields. Productive breeding sites were characterized by stable micro-climatic conditions. Species composition of emerging sand flies varied with habitat, season and geographical location.

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.

Studies on the flight patterns of foraging sand flies.

Phlebotomine sand flies transmit Leishmania parasites that cause leishmaniasis in humans. We report experimental results that improve our understanding of how foraging sand flies proceed over flat or sloping ground and how they negotiate vertical obstacles. Three rows of traps were suspended at different heights on a wire fence. Those just above ground level captured 87% of all flies, traps set at one meter captured 11% while only 2% of the flies were captured in traps set two meters above ground. When traps were deployed on a vertical support wall, the mean catch per trap was much higher than for traps suspended on the fence. Traps suspended just above ground level captured 57% of all flies, traps set at one meter above ground captured 27% of the flies and even traps set at two meters captured 16% of the flies. Although, most flies were still captured close to the ground, a higher percentage reached the second and third rows of traps. Sticky traps on a vertical wall produced similar results with significantly more flies alighting on the lower sections of the trap closest to the ground. On a vertical sand fly-proof net the overall dispersal of the flies was more like on a wall than in open space. Traps suspended just above ground level captured 49%, traps set at one meter above ground captured 36% and traps set at two meters captured 15% of the flies. Following spraying of the net with deltamethrin (1%), fewer sand flies were captured but the reduction was not statistically significant. Our conclusions are that being small and frail, sand flies tend to fly close to the ground probably in order to avoid being swept away by gusts of wind. When they encounter a vertical obstacle, they proceed upwards close to the obstacle with intermittent stops. Therefore, insecticide-sprayed walls or vertical nets should be effective for controlling sand flies approaching human habitation.

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.

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.