Sand flies used to have a reputation for being difficult and labour-intensive to breed. Here we summarize our experience with establishment and maintenance of sand fly colonies and their use for infective experiments: techniques for collection and handling wild-caught females, rearing larvae and adults and experimental infections of sand flies by Leishmania using membrane feeding. In addition, we compare major life cycle parameters between various colonies maintained under standard laboratory conditions. The sand fly rearing is tricky but some species can be reared in large numbers with a minimum of space and equipment. Initiation of new colonies from endemic sites is a prerequisite for accurate studies on parasite-vector interaction but it is more difficult step than routine maintenance of colonies already established in laboratory for many generations.

The control of the sand fly vectors of leishmaniasis is problematic because their larvae develop in largely unknown terrestrial habitats making them impervious to available control measures. Furthermore, the behavior patterns of adults of different sand fly species are highly diverse, requiring tailor-made control solutions based upon a profound knowledge of their biology. In this short review, we describe possible lines of research that hold promise for improving our munitions in the battle against the diseases they transmit. The suggested approaches are not necessarily presented in order of importance, but rather in a logical sequence starting in the larval breeding areas where the sand flies originate and culminating with the human environments. Some examples are offered to illustrate the potential efficacy.

I review species concepts, the taxonomy of phlebotomine sand flies, and some transmission cycles of leishmaniasis in order to illustrate the difficulties of classifying these vectors in a way that will be ideal both for medical parasitologists and sand fly specialists. Choices will have to be made between different classifications, either maintaining a practical one containing few vectorial genera (mostly Phlebotomus for the Old World and Lutzomyia for the Neotropics) or changing the generic names of many vectors so that the classification represents an evolutionary hypothesis. However, sand flies also transmit arboviruses and members of other sand fly genera bite humans, and so vectorial status alone might not provide the criteria for recognizing only a few genera. Vectorial roles are often determined by species-level co-evolution of susceptibility to Leishmania species, with selection being initiated and maintained by ecological contacts. There is only imperfect cocladogenesis of genus-level groups or subgeneric complexes of sand flies and Leishmania species. Natural hybridization between sand fly species has been recorded in several species complexes, and this highlights the need to focus on gene flow and the distribution of phenotypes of biomedical importance, not on taxa.

A literature review is provided on the state of knowledge of the ecology and control of the sand fly vectors of Leishmania donovani in East Africa, with a special emphasis on Phlebotomus orientalis. Visceral leishmaniasis caused by L. donovani is a major health problem in several areas in East Africa. Studies conducted in the past 70 years identified P. orientalis Parrot and P. martini Parrot as the principal vectors of L. donovani in Sudan, Ethiopia and Kenya and P. celiae Minter as the secondary vector of the parasite in one focus in Ethiopia. Findings on sand fly fauna and other circumstantial evidence indicate that P. martini is also responsible for transmission of L. donovani in VL endemic foci of Somalia and Uganda. Several studies showed that P. orientalis occupy distinct habitat characterized by black cotton soil and Acacia seyal-Balanites aegyptiaca vegetation, whereas P. martini and P. celiae are associated with termite mounds. Little knowledge exists on effective control measures of sand fly vectors of L. donovani in East Africa. However, recent evidence showed that use of insecticide impregnated bednets and insect repellents may reduce exposure to the bites of P. orientalis.

The object of this study was to determine the genetic structures of three vector species, Phlebotomus tobbi, Phlebotomus papatasi, and Phlebotomus sergenti, in the Cukurova Region of Turkey, an endemic focus of cutaneous leishmaniasis. The genetic diversity indices, neutrality tests and hierarchical analysis of molecular variance (AMOVA) were performed using partial sequences of ITS2 and cytochrome b gene regions. In all species, within population genetic variation was higher than between population variation for ITS2 gene region. Fst values were low and non-significant for P. sergenti, and were higher for P. papatasi and P. tobbi indicating a weak structuring between populations. AMOVA tests suggest any substantial isolation between populations within species. AMOVA analysis of cyt b gene region revealed significant genetic structuring between populations for P. papatasi and P. sergenti. Fst values were relatively high and significant for these species indicating a certain degree of isolation between populations. However, in P. tobbi, any significant population genetic structuring was detected. Tajima's D and Fu's Fs values were negative and significant in all three species might be indicating a demographic expansion.

An intraspecific study of Phlebotomus sergenti was performed on populations from Turkey, Syria, Israel, and Uzbekistan by four different approaches: geometric morphometries, RAPD analysis, internal transcribed spacer 2 (ITS2) sequencing (nuclear marker), and cytochrome B sequencing (mitochondrial marker). In RAPD analysis, distinct clades were formed in accordance with the geographical origin of the specimens. There was no distinct grouping according to place of origin within the Turkish samples from various localities in south-eastern Anatolia, which suggests a gene flow between populations separated spatially by the Amanos mountains, a mountain range of a considerable altitude. The results of ITS2 rDNA sequencing complied with the previously published intraspecific division of P. sergenti into two branches, northeastern and southwestern. However, mtDNA haplotypes formed three lineages with specimens from Turkey and Israel, sharing a common clade. A previously postulated hypothesis about a complex of sibling species within P. sergenti is therefore questionable. Cytochrome B seems to be a more discriminative marker for intraspecific variability assessment.

Sand fly saliva contains an array of bioactive molecules that facilitate blood feeding and also function as modulators of the vertebrate immune response. Such a complex of biologically active molecules was shown to be both conserved and divergent among sand fly species. It is likely that expression of sand fly salivary molecules could be modulated by environmental factors, both biotic and abiotic, that ultimately dictate the quality, and possibly quantity, of the secreted saliva. Carbohydrates are an integral part of the sand fly diet, and sugar-sources found in natural habitats are potentially involved in defining the profile of sand fly saliva, and may influence vectorial capacity. Saliva can drive the outcome of Leishmania infection in animal models, and salivary molecules are potential targets for development of vaccines to control Leishmania infection. Thus, identifying what environmental factors effectively modulate sand fly saliva in the field is a critical step towards the development of meaningful protection strategies against leishmaniasis that are based on salivary compounds from sand fly vectors.

We have previously shown that fermented ripe fruit is a strong attractant for several mosquito species, and when mixed with oral insecticide these attractive toxic sugar baits (ATSB) were highly effective for local mosquito control. In the present study we compared the effects of ATSB presented in different ways on isolated populations of Phlebotomus papatasi Scopoli. Experiments were carried out in the arid habitat of the Jordan valley, Israel where the effectiveness of three methods was compared: ATSB sprayed on patches of vegetation, net fence coated with ATSB, and bait stations soaked with ATSB. Spraying ATSB reduced the population to about 5% of the control area population. Barrier ATSB coated fences, had a similar effect decreasing the population to about 12% of the concurrent catch in the control site. The effect of ATSB presented on bait stations was much smaller and compared to the control, only caused the population to be reduced to 40%. In the control areas where only food dye marker was used, the solution presented on bait stations only marked an average of 22.3% of female sand flies while spraying vegetation and using barrier fences in the two other experiments marked about 60% of the females. Our experiments show that ATSB either sprayed on the vegetation or on barrier fences is an effective means against sand flies at least in arid areas where attractive plants are scarce or absent.

Sand fly populations of different ecological niches in the Amaraji endemic American Cutaneous Leishmaniasis (ACL) focus of the Pernambuco Atlantic Forest region of northeastern Brazil were monitored spatiotemporally. Lutzomyia whitmani was dominant in all niches but occurred in smaller numbers in forested locations. L. whitmani was significantly less seasonal than the other species, being present throughout the year while other species were more abundant between February and April. These results suggest that L. whitmani may potentially be the principal vector of ACL in the region, even though the sand fly fauna was diverse: 88% were L.whitmani and 12% belonged to 11 other species. Two other species, L. complexa (1.3%) and L. migonei (0.8%), considered to be ACL vectors in other regions, were also present. This detailed picture of the sand fly populations abundance and spatiotemporal distribution provides a basis for future modeling studies of forecasting sand fly activity patterns and ACL occurence.

Human indigenous cutaneous leishmaniasis caused by Leishmania donovani complex is endemic in Sri Lanka. We performed an entomological survey to determine the distribution of probable vector species. Sand flies were collected in districts in the dry zone, in the wet zone highlands, and in the wet zone coastal belt of Sri Lanka using CDC light traps, sticky traps and cattle-baited net traps during July, 2005. The survey was reconducted in February, 2006. Overall, 584 sand flies belonging to Phlebotomus (266 specimens, 2 species) and Sergentomyia (318 specimens, 8 species) genera were collected. A total of 266 Phlebotomus was identified as P. argentipes (258/266; 97%) and P. stantoni (8/266; 3%). The identification studies of Sergentomyia specimens showed that there are at least 8 species in Sri Lanka. Higher number of Phlebotomus sand flies (76/266) were caught in the southern part of the country compared to the other parts probably due to different ecological aspects. P. argentipes were widely distributed throughout the island whereas P. stantoni were collected only in four districts. Since P. argentipes is known to be the vector of L. donovani responsible of visceral leishmaniasis in India, this species may be incriminated as the most possible vector of human cutaneous leishmaniasis in Sri Lanka.

This paper presents the results of an entomological survey in an endemic focus of cutaneous leishmaniasis in the Cukurova region of Turkey. A total of 8,927 specimens belonging to eight Phlebotomus and two Sergentomyia species were captured with sticky papers and CDC light traps from 52 stations. Phlebotomus tobbi Adler, was found to be the most abundant species. Sand fly activity started in May and ended in October. Abundance was highest in August. According to the frequency distributions among certain temperature intervals the observed number of individuals was significantly different from the expected values between 22–24° C and 28–30° C. There was no significant correlation between the abundance of sand flies and altitude. However, sand fly species showed great aggregation at the 100–199 m and 200–299 m altitude intervals. The Shannon—Weinner index indicated no difference between the diversity and abundance of sand flies at different altitudes. Diversity and evenness reached maximum values at 500 m. Jaccard's coefficient indicated that similarity was the highest between 0–99 and 300–399, 0–99 and 500–599 and 100–199 and 200–299 m and lowest between 100–199 and 300–399 and 100–199 and 500–599 m.

Cutaneous leishmaniasis, caused by Leishmania (Viannia) braziliensis, is sporadic in many rural and suburban areas of Rio de Janeiro State. An investigation was carried out during 2008/9 in the Municipality of Saquarema, Rio de Janeiro, Southeast Brazil, in order to identify the phlebotomine sand fly fauna. More than 2,100 sand flies were collected in peridomestic areas in two chicken coops using CDC light traps. Nine species of phlebotomine sand flies were identified: Nyssomyia intermedia, Nyssomyia whitmani, Pintomyia (P.) pessoai, Pintomyia (P.) fischeri, Pintomyia (P.) bianchigalatiae, Migonemyia (M.) migonei, Lutzomyia (L.) longipalpis, Brumptomyia cunhai and Brumptomyia guimaraesi. Based on the results of this study together with related studies in other CL foci in Rio de Janeiro, both Nissomyia intermedia and Migonemyia migonei can be considered suspect vectors of the disease in the region. The potential risk of VL due to the presence of its proven vector L. longipalpis is discussed.

An entomological survey was conducted to determine the spatial distribution of phlebotomine fauna and understand the effect of environmental factors. The entomological survey was carried out during 2006–2007 in a study area in the rural area of Aydin province, near the Kusadasi town where VL, CL, and canine leishmaniasis (CanL) are endemic. In 2006 and 2007, 132 locations were sampled using sticky traps mainly on embankments. Detailed environmental and meteorological information was also collected for each location. The results of entomological studies indicated that the probable vectors are Phlebotomus tobbi and P. neglectus for VL and CanL, and P. similis for CL in this western leishmaniasis focus. The data revealed a correlation between their presence and spatial variables such as altitude, sampling site location, and humidity. The distribution areas of probable vector species in this study area allowed the identification of risk levels, which may provide useful information to guide the leishmaniasis research in endemic regions.

This study examined the spatial distribution and seasonal fluctuations of population densities of phlebotomine sand flies and was designed to obtain baseline data on the population trends of Phlebotomus argentipes, P. papatasi, and Sergentomyia spp. in a visceral leishmaniasis endemic area of Bihar, India. Beginning on 28 October 2009 and through 20 October 2010, 63 CDC light traps were evenly distributed in human homes, cattle sheds, combined dwellings, chicken coops, and adjacent vegetation areas in three villages in the Saran District of Bihar State. Sand fly collections were made on a weekly basis, sorted, and identified according to species, sex, and feeding status of the two genera. The daily temperatures and relative humidity ranges were collected in a representative human home, cattle shed, and combined dwelling in each of the three study villages. Village census surveys were conducted in the three study villages in February 2010, acquiring human population data, structural composition data, and livestock census information, and documenting the history of visceral leishmaniasis within each household. A total of 52,653 sand flies was trapped and identified over 3,276 trap-nights. Peaks in abundance were observed in November 2009, March and April, June through August. Of the sand flies trapped, 72.1% were P. argentipes, 27.1% Sergentomyia spp., and 0.8% P. papatasi. Distribution of the sand fly captures included 30.6%, 26.7%, 18.6%, 12.1%, and 12.0% from vegetation, combined dwellings, cattle sheds, housing, and poultry houses, respectively.

Altogether, 4,008 sand flies belonging to seven species were collected over a period of one year in the microhabitats of a single canyon in the Carmel Mountain ridge. The three most abundant were P. arabicus, P. tobbi, and P. simici. Our results suggest that none of the seven sand fly species was indifferent to the heterogeneity of the microenvironment inside the canyon. Apart from the rare P. perfiliewi, which was only collected on the upper part of the south-facing slope, and P. tobbi, which clustered on the north-facing slope, the bulk of the other sand flies were caught on the bottom of the canyon. During the summer, the catches of all sand fly species increased to reach their maximum number in August and September. In April and May, there was lush vegetation and humidity, so species were distributed evenly throughout their habitats. With the onset of summer dryness, the sand flies concentrated in the humid habitats. The rate of concentration was essentially higher for males than for females, and this variation may result from differences in the behavior of the two sexes. During our study, none of the 2,318 dissected female sand flies were positive for Leishmania promastigotes.

Monitoring sand flies in the cutaneous leishmaniasis foci Kfar Adummim and Ma'ale Adummim from May to October generally yielded several hundred specimens per CO₂ baited trap. In the summer of 2009, a sharp rise in the number of sand flies trapped was recorded in Kfar Adummim, while numbers were similar to previous years in Ma'ale Adummim; approximately 4,000 specimens compared to about 400, with maximal catches of about 16,500 specimens in Kfar Adummim. We postulate that the sharp increase in sand fly numbers is directly related to the intensive construction conducted which enhanced sand fly breeding habitats.

In laboratory studies, insecticides (diflubenzuron, novaluron, methoprene and, pyriproxyfen) that have been incorporated into rodent diets were effective as feed-throughs against sand fly larvae. Novaluron also was effective against sand fly larvae at low concentrations and under simulated field conditions. Ivermectin has been shown to be effective as a systemic insecticide, killing 100% of blood-feeding sand flies for up to seven d after rodents were treated. The fluorescent tracer technique (FTT) is the use of certain fluorescent dyes (rhodamine B or uranine O) as feed-through transtadial biomarkers for phlebotomine sand flies, systemic biomarkers for blood-feeding sand flies, and permanent markers for nectar-feeding sand flies. The results of these laboratory studies provide proof of concept for the FTT and indicate that the FTT could be used to delineate specific foci with rodent/sand fly associations that would be susceptible to control by using feed-through or systemic insecticides, or foci where insecticide-treated sugar baits could be used against sand flies.

Oviposition behavior is a fairly neglected aspect in our understanding of the biology of sand flies. In this study, we used a comparative approach using both new- and old-world species (Lutzomyia longipalpis and Phlebotomus papatasi) in choice and no-choice oviposition chambers to evaluate the effect of old sand fly colony remains (frass), conspecific eggs, and their combination on oviposition rates of these sand flies. We also tested the effect of egg washing with de-ionized water on oviposition rates. In both choice and no-choice experiments, sand fly species laid more eggs on a substrate containing frass. The effect of eggs alone was not significant but showed a positive trend. Furthermore, for both sand fly species, the effect of the combined treatment was sub-additive suggesting a potential inhibitory effect of one factor on the other. Egg washing did not have a significant effect. The choice and no-choice experimental designs did not differ in their outcomes suggesting the choice-design could serve as an effective high throughput method for screening oviposition attractants/stimulants.

In this study, we tested the capacity of Temperature Gradient Gel Electrophoresis (TGGE)-based fingerprinting of 16S rDNA PCR fragments to assess bacterial composition in a single isolated sand fly gut. Bacterial content was studied in different life stages of a laboratory-reared colony of Phlebotomus duboscqi and in a wild-caught Phlebotomus papatasi population. Our study demonstrates that a major reorganization in the gut bacterial community occurs during metamorphosis of sand flies. Chloroflexi spp. was dominant in the guts of pre-imaginal stages, although Microbacterium spp. and another as yet unidentified bacteria were detected in the gut of the adult specimen. Interestingly, Microbacterium spp. was also found in all the adult guts of both species. We demonstrate that the analysis of bacterial diversity in an individualized sand fly gut is possible with fingerprinting of 16S rDNA. The use of such methodology, in conjunction with other culture-based methods, will be of great help in investigating the behavior of the Leishmania-bacterial community in an ecological context.

Our goal was to study the effectiveness of the insecticide imidacloprid as a systemic control agent. First, to evaluate the blood-feeding effect, we fed adult female Phlebotomus papatasi with imidacloprid-treated rabbit blood and monitored blood-feeding success and survival. Second, to evaluate the feed-through effectiveness of this insecticide, we fed laboratory rats and sand rats with insecticide-treated food and evaluated the survival of sand fly larvae feeding on rodents' feces. In the blood-feeding experiment, 89.8% mortality was observed with the higher dose (5 mg/ml) and 81.3% with the lower dose (1 mg/ml). In the larvicide experiments, both sand fly species demonstrated a typical dose-response curve with the strongest lethal effect for the 250 ppm samples. Lutzomyia longipalpis larvae, however, were less sensitive. In all experiments, 1^st instar larvae were more sensitive than the older stages. First instar P. papatasi larvae feeding on sand rat feces passed the larvicidal threshold of 90% mortality at doses higher than 50 ppm. In comparison, in older stages 90% mortality was obtained with a dose of only 250 ppm. Overall, results support the feasibility of imidacloprid as a systemic control agent that takes advantage of the tight ecological association between the reservoir host and the sand fly vector.

The OFF! Clip-On fan vaporizer device releasing metofluthrin was evaluated against phletobomine sand flies in the Judean Desert, Israel, in October, 2009. A total of 76,400 sand flies was collected, with male flies representing 98.3% Phlebotomus sergenti and 1.7% P. papatasi. Females comprised 43.0% of the total catch and included 6.7% blood-fed females. Similar proportions of flies were collected in both suction and sticky traps. In trials with unbaited suction traps, similar numbers of sand flies were collected in traps with a metofluthrin device, blank device, or no device (i.e., suction only). In suction traps baited with CO₂, higher numbers of P. sergenti males and blood-fed females were collected in traps with a blank device compared to traps with a metofluthrin device. In sticky traps baited with CO₂, there were no significant differences between catches in traps with a metofluthrin device, blank device, or no device. The results suggest metofluthrin from the device is not repellent against sand flies in a field environment despite showing insecticidal activity against flies collected in suction traps.

In this study, we evaluated the efficacy of eleven commercial models of propane combustion traps for catching male and female Phlebotomus papatasi. The traps differed in physical appearance, amount of carbon dioxide produced and released, type and location of capturing device, and the method by which the trap suction fans were powered. The traps tested were the Mosquito MagnetTM(MM)-Pro, MM-Liberty, MM-Liberty Plus, MM-Defender, SkeeterVac®(SV)-35, SV-27, Mosquito DeletoTM(MD)-2200, MD-2500, MT150-Power Trap, and two models of The Guardian Mosquito Traps (MK-01 and MK-12). All trap models except the SV-35, the SV-27, the MD-2500, and the MK-12 attracted significantly more females than males. The SV-35 was the most efficient trap, catching significantly more females than all the other models. The MD-2200 and MK-12 models were the least effective in catching either female or male sand flies. These data indicate that several models of propane combustion traps might be suitable substitutes for either CO₂-baited or unbaited light traps for adult sand fly surveillance tools. One advantageous feature is the traps' ability to remain operational 24/7 for ca. 20 days on a single tank of propane. Additionally, the models that produce their own electricity to power the trap's fans have an important logistical advantage in field operations over light traps, which require daily battery exchange and charging.

We tested the performance of ten commercial mosquito traps with varying attractive features, against three CDC traps (an unlit model 512, an incandescently lit model 512, and a UV lit model 1212) as well as simple sticky paper, for their ability to attract and capture Phlebotomus papatasi in Israel. The commercial traps tested were the Sentinel 360, the Combo Trap, the Mega Catch Premier, the Bug Eater, the EcoTrap, the Galaxie Power-Vac, the Biter Fighter, the Black Hole, the Mosquito Trap, the Mosquito Catcher, the Sonic Web, the Solar Pest Killer, and a Bug Zapper. The four best performing traps with the highest nightly catches were the Sentinel 360 (85.96±19.34), the Combo Trap (70.00±7.78), the Mega Catch Premier (51.93±1.82) and the UV lit CDC 1212 trap (47.64±3.43). Five traps, the Mosquito Trap, the Mosquito Catcher, the Sonic Web, the Solar Pest Killer, and the Bug Zapper, performed exceptionally poorly, catching an average of less than two sand flies per day. To our knowledge, this is the first comprehensive attempt to evaluate commercial traps for their effectiveness in catching sand flies, and we show here that some traps that have been effective in catching mosquitoes are also effective in catching sand flies.

Knowledge about diurnal resting sites of sand flies is scanty and often anecdotal. In this study, we explored a part natural - part agricultural oasis in Neot Hakikar, Israel, looking for sand fly resting sites. To achieve this, we developed a new type of emergence trap. Sixteen types of microhabitats were examined and in seven of these, we also investigated the rodent burrows. We found that Phlebotomus papatasi showed clear preferences for resting sites characterized by vegetation cover, type of vegetation, and the presence of a mulch layer. In habitats with bare soil and little shade, few or no resting sand flies were found outside rodent burrows. Apart from the trunks of date trees, most resting P. papatasi were found in disturbed habitats, especially in large piles of organic waste and in a plowed field. Though catches from rodent burrow exits were always higher than from the nearby ground, it is safe to assume that the few burrows in this vast oasis do not play an important role for breeding and resting of P. papatasi. It also appears that disturbing the natural environment further increases the already considerable sand fly population.

Sugar is the main source of energy for the daily activities of sand flies. Considering its importance, there is surprisingly little information on sugar meal specific sources and sand fly attraction to plants, particularly in the field. In this study, we first needed to develop an effective sand fly trap that would be suitable for mass screening of potentially attractive flowering plants. Next, we used this trap to screen a total of 56 different flowering plant species and five plant species soiled with different types of honeydew. The plant baited traps together caught 21,978 P. papatasi. Out of the 56 types of flowering plants which were tested, 13 were shown to bait significantly more female sand flies, and 11 baited more male sand flies than the control. Based on an attraction index, the top three attractive plants in this study were the flowering plants Ochradenus baccatus, Prosopis farcta, and Tamirix nilotica. We believe that plants and phyto-chemicals have untapped potentials to attract sand flies. These could be used for control and, in combination with simple glue traps, as an alternative for existing monitoring systems.

Recently, in several areas of the Middle East, a sharp increase of cutaneous leishmaniasis was observed in suburbs of larger towns including Jerusalem. In some of these areas, poor housing conditions and unsuitable waste management was suspected to provide ideal conditions for sand fly breeding, but hard data on diurnal resting sites and breeding habitats of most sand fly species are scant. In this study, we chose 16 sites on both slopes and the bottom of a natural valley in the Judean Desert to conduct a survey of sand fly distribution with emergence traps. Altogether, 1,261 sand flies, 52% Phlebotomus syriacus, 22% P. sergenti, 14% P. papatasi and 12% P. tobbi were caught. About two thirds of the flies caught were resting, while the other third emerged from breeding sites. All four species showed clear preferences for resting and breeding sites, but generally, most sand flies were breeding in the more humid habitats, namely the bottom of the valley, the adjacent north facing slope, terraces on the north facing slope, and caves. The vegetation cover also appeared to be important for resting habitats; on the bottom of the valley more than six times as many sand flies were collected in areas covered by dense vegetation than in areas with low vegetation cover. P. sergenti seemed also to better tolerate the drier habitats, which might explain the abundance of this species in the arid Judean Desert.

Sand flies have been reported feeding on various plant organs including stems, leaves, and flowers but the attraction of sand flies to sugar-rich fruits has received little attention. In this study, we tested 24 commercially available fruits for their attractiveness to sand flies, and found that the top three attractive fruits were nectarine (Prunus persica var. nectarina), cactus fruit, (Opuntia ficus-indica), and guava (Psidium guajava). These fruits were fed upon equally by both males and females. There were slight differences in the order of preference to the less-attractive fruits by males and females, but these were not statistically significant. The knowledge of fruit preference may help to improve existing methods that use plant phytochemicals to attract and kill biting flies.

We conducted two experiments to determine the best CDC-trap configuration for catching male and female Phlebotomus papatasi. First, visual features were evaluated. Standard CDC traps were modified to have black or white catch bags, black or white lids, or no lids and these were tried in different combinations. Significantly more male sand flies were caught by darker traps; significantly more females were captured by traps with either all black or a combination of black and white features. Attraction may be due to dark color or contrast in colors. CDC traps with suction and the following features were also evaluated: no light; incandescent light; ultraviolet (UV) light; combination of black color, heat and moisture; CO₂ alone, or a combination of black color, heat, moisture, and CO₂ simultaneously, all in upright and inverted positions, with the opening for insect entry always 50 cm above the ground. Significantly more females than males were caught by all traps (standard and inverted) except the control traps with suction only. Traps with CO₂ caught more sand flies than traps without CO₂ Traps with black color, heat and moisture captured significantly more sand flies than the control traps, but with the addition of CO₂, these traps catch significantly more sand flies than the other traps evaluated. Inverting traps increased the catch for like traps by about two times.