Natural breeding places of phlebotomine sandflies

Methods of finding larvae and pupae of phlebotomine sandflies (Diptera: Psychodidae) are described and the known types of breeding sites used by sandflies are listed. Three ways of detecting sandfly breeding places are the use of emergence traps placed over potential sources to catch newly emerged adult sandflies; flotation of larvae and pupae from soil, etc., and desiccation of media to drive out the larvae. Even so, remarkably little information is available on the ecology of the developmental stages of sandflies, despite their importance as vectors of Leishmania, Bartonella and phleboviruses affecting humans and other vertebrates in warmers parts of the world. Regarding the proven or suspected vectors of leishmaniases, information on breeding sites is available for only 15 out of 29 species of sandflies involved in the Old World and 12 out of 44 species of sandflies involved in the Americas, representing approximately 3% of the known species of Phlebotominae. Ecotopes occupied by immature phlebotomines are usually organically rich moist soils, such as the rain forest floor (Lutzomyia intermedia, Lu. umbratilis, Lu. whitmani in the Amazon; Lu. gomezi, Lu. panamensis, Lu. trapidoi in Panama), or contaminated soil of animal shelters (Lu. longipalpis s.l. in South America, Phlebotomus argentipes in India; P. chinensis in China; P. ariasi, P. perfiliewi, P. perniciosus in Europe). Developmental stages of some species (P. langeroni and P. martini in Africa; P. papatasi in Eurasia; Lu. longipalpis s.l. in South America), have been found in a wide range of ecotopes, and many species of sandflies employ rodent burrows as breeding sites, although the importance of this niche is unclear. Larvae of some phlebotomines have been found in what appear to be specialized niches such as Lu. ovallesi on buttress roots of trees in Panama; P. celiae in termite hills in Kenya; P. longipes and P. pedifer in caves and among rocks in East Africa. Old World species found as immatures in the earthen floor of human habitations include P. argentipes, P. chinensis, P. martini and P. papatasi. Much more information on sandfly breeding sites is required to facilitate their control by source reduction.     

The roles of Phlebotomus martini and P. celiae (Diptera: Phlebotominae) as vectors of visceral leishmaniasis in the Aba Roba focus, southern Ethiopia

Abstract. During field studies (December 1988 to April 1990) to determine the vectors of visceral leishmaniasis (kala-azar) in the Aba Roba (Segen Valley) focus of southern Ethiopia, a total of 40,770 sandflies (Diptera: Psychodidae: Phlebotominae) were collected and identified as six species of Phlebotomus and seventeen Sergentomyia spp. Nine of these species were anthropophilic (four Phlebotomus and five Sergentomyia spp.), the dominant being Phlebotomus (Synphlebotomus) martini, P.(S.)celiae and Sergentomyia (Sergentomyia) schwetzi. P. celiae adult populations increased during the rainy season. Of 2326 P. martini and 1044 P. celiae dissected, the parous rate was 29.9% and 24.4%, respectively. Leishmania promastigotes were detected in sixteen P. martini (eleven identified as L. donovani by isoenzyme analysis and/or a DNA probe) and in three P. celiae (two identified by DNA probe), giving overall infection rates of 0.7% (1.9% parous infection rate) and 0.3% (1.2% parous infection rate), respectively. Four isolates from P. martini were typed by their isoenzyme profile as L. donovani zymodeme MON-37 = LON-44, identical to one of the zymodemes isolated from patients in this focus. This is the first evidence that P. celiae is a vector and the first time that P. martini has been implicated in Ethiopia. Based on observations of abundance, seasonality and prevalence of infection of the two Synphlebotomus spp., it is concluded that P. martini is the major vector and P. celiae a secondary vector in the Aba Roba focus. The risk of infection with L. donovani is greatest during the wet season.     

Host feeding preference of Phlebotomus guggisbergi,a vector of Leishmania tropica in Kenya

Abstract. Recently the sandfly Phlebotomus guggisbergi was found to be a vector of Leishmania tropica causing cutaneous leishmaniasis in the Laikipia focus, Kenya, but extensive searches have shed no light on the identity of the rural reservoir host(s). In order to discover more about the biology of the vector, a host feeding preference study was conducted on wild sandflies in their natural cave environment over a 6-month period. Solid state Army miniature (SSAM) traps, without light bulb, were suspended over cages with potential hosts or an empty cage control. The animals tested included sheep, goat, dog, cat, hamster, rabbit, giant rat (Cricetomys gambianus), crested rat (Lophiomys imhausi) and rock hyrax (Procavia capensis), all of which (except hamsters) are normally found in the vicinity of the study site. Sandfly collections from traps baited with goat, sheep, cat, dog, rabbit, or wild rodent species were significantly higher than the control, whereas trap collections with hamster and rock hyrax were not significantly different from the control. Numbers of sandflies collected from the goat, sheep and cat were significantly greater than from the rabbit and rodents. The sex ratio also varied between collections: larger animals attracted a higher proportion of female P.guggisbergi than did the smaller animals (P>0.05). Therefore greater emphasis should be placed on surveying larger animals to assess their status as reservoir hosts for L.tropica in Kenya.     

Aspects of the bionomics of Phlebotomus sergenti sandflies from an endemic area of anthroponotic cutaneous leishmaniasis in Aleppo Governorate, Syria

Aspects of the bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied from June to November 2005 in three foci of anthroponotic cutaneous leishmaniasis (ACL) in Aleppo Governorate, Syria, where the agent Leishmania tropica (Wright) (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus sergenti Parrot. Syria has been designated by the World Health Organization as one of four countries in the Old World where cutaneous leishmaniasis is hyperendemic, but little is known about the biology of local vector populations. Standard collections by sticky traps showed two peaks in density, in June and late August. In total, 1840 sandflies were caught, comprising five species: Phlebotomus papatasi (Scopoli) (68.0%); P. sergenti (25.4%); Sergentomyia minuta (Rondani) (6.4%); Phlebotomus tobbi Adler & Theodor (0.1%), and Phlebotomus mascittii canaaniticus Adler & Theodor (0.1%). Similar numbers of P. sergenti were caught indoors (246 specimens) and outdoors (222), whereas P. papatasi was significantly more abundant indoors (1096 specimens) than outdoors (156) (χ² = 241, P < 0.01). In total, 212 blood-fed females were tested for host blood determination, of which 176 (83.0%) reacted with anti-species reagent. Results from 20 P. sergenti suggest that this species is an opportunistic feeder, imbibing human, ovine, avian, bovine and feline blood, although more bloodmeals were taken from humans and cattle than expected in relation to the relative proportions of potential hosts present (the forage ratio, FR). The bionomics of P. sergenti are discussed in relation to the inefficacy of control campaigns based on indoor spraying with residual insecticides that have been implemented by the Syrian Ministry of Health to control the epidemics of ACL in the Aleppo Governorate.     

Phlebotomine sandflies (Diptera-Psychodidae) of the isle of Cyprus. II--Isolation and typing of Leishmania (Leishmania infantum Nicolle, 1908 (zymodeme MON 1) from Phlebotomus (Larroussius) tobbi Adler and Theodor, 1930

During two surveys conducted in Cyprus (August 1998 and September 1999), 2,910 phlebotomine sandflies females were caught by CDC miniature light traps then dissected under binocular and examined on microscope. Eleven species were identified: Phlebotomus papatasi, P. sergenti, P. jacusieli, P. alexandri, P. tobbi, P. galilaeus, P. mascittii, P. economidesi, Sergentomyia fallax, S. minuta et S. azizi. The Larroussius species (P. galilaeus and P. tobbi) are the most abundant (more than 60% of our captures). Promastigotes were isolated from one specimen identified as P. tobbi. A Leishmania stock was successfully cultured and identified by isoenzyme characterisation as belonging to L. infantum zymodeme MON 1. The same zymodeme was isolated and identified from four dogs too. Because of the absence of usual vectors of L. infantum in the eastern part of the Mediterranean basin (P. neglectus and P. syriacus), and according to its distribution in Cyprus, P. tobbi constitute certainly a good local vector. It seems to be not very anthropophilic, that could explain the very few human cases.     

The efficacy of indoor CDC light traps for collecting the sandfly Phlebotomus argentipes, vector of Leishmania donovani

The efficacy of light traps for collecting sandflies (Diptera: Psychodidae) varies both inter-specifically and intra-specifically (by gender and physiological status) as a result of significant differences in phototropic and other behavioural characteristics. The efficacy of miniature CDC light traps for collecting Phlebotomus argentipes Annandale & Brunetti, a vector of Leishmania donovani Laveran & Mesnil (Kinetoplastida: Trypanosomatidae), was assessed in the Indian state of Bihar. Sandflies were collected during the night from 16 houses in each of three villages over 3 months (four times at fortnightly intervals) using CDC light traps indoors, and by aspirator collection (carried out by one person for 30 min/house) from the walls of the same houses the following morning. Incidence rate ratios (IRRs) between CDC light trap collections and aspirator collections were obtained through a negative binomial regression with household as random effect. CDC light traps were especially effective in catching males (IRR 3.08, 95% confidence interval [CI] 2.12-4.46) and unfed females (IRR 3.50, 95% CI 2.37-5.16) of P. argentipes, and to a lesser extent gravids (IRR 1.77, 95% CI 1.07-2.93). However, only a relatively small proportion of all blood-fed P. argentipes were collected by light trap (IRR 0.45, 95% CI 0.28-0.73). Despite its limitations in collecting blood-fed female sandflies, the CDC light trap appears to trap a sufficient proportion of the indoor population of sandflies for sampling purposes, and as this light trap is also more convenient and more easily standardized than the aspirator method, we conclude that it is the most efficient method for monitoring P. argentipes populations in the Indian subcontinent.     

Hosts of Lutzomyia shannoni (Diptera: Psychodidae) in relation to vesicular stomatitis virus on Ossabaw Island, Georgia, U.S.A.

Abstract. Hosts of Lutzomyia shannoni Dyar, a suspected biological vector of the New Jersey serotype of vesicular stomatitis (VSNJ) virus, were determined using an indirect enzyme-linked immunosorbent assay (ELISA) of 333 blood-fed female sandflies collected from their diurnal resting shelters on Ossabaw Island, Georgia, U.S.A. Sandflies had fed primarily on white-tailed deer ( Odocoileus virginianus ) (81%) and to a lesser extent on feral swine ( Sus scrofa ) (16%), two species of host infected annually with VSNJ. Other hosts were raccoons ( Procyon lotor ) and horses ( Equus caballus ) or donkeys ( E. asinus ), with only two (<1%) mixed bloodmeals from deer/raccoon and deer/swine. A larger proportion of feedings on feral swine was detected in maritime live oak forests than in mixed hardwood forests. These findings are consistent with the hypothesis that L. shannoni is a primary vector of VSNJ virus on Ossabaw Island.     

Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis

We developed a DNA assay for bloodmeal identification in haematophagous insects. Specific host cytochrome B gene sequences were amplified by PCR and classified on the basis of their mobility in a heteroduplex assay. In the blackfly Simulium damnosum s.l. (Diptera: Simuliidae), human cytochrome B DNA sequences were identifiable up to 3 days following ingestion of the bloodmeal. In the tsetse Glossina palpalis (Diptera: Glossinidae) collected from tsetse traps in Ivory Coast, bloodmeals were identified as taken from domestic pigs on the basis of their heteroduplex pattern and DNA sequence. Evidently the cytochrome B sequence shows sufficient interspecific variation to distinguish between mammalian host samples, while exhibiting minimal intraspecific variation. The stability of DNA in bloodmeals, for several days post-ingestion by haematophagous insects, allows PCR-HDA assays to be used reliably for host identification.     

Aspects of sandfly biology in southern Switzerland

Abstract. During three field surveys of phlebotomine sandflies in the region of Sessa, Minor Malcantone, southern Switzerland, annually 1987-89, Phlebotomus pemiciosus and P.mascittii were caught regularly by means of light traps, but only when the traps were in close association with humans. The most productive catching sites were in the middle of Sessa village, in basements or partially closed shelters in the immediate vicinity of human dwellings. All catching sites outside the village were negative. The only other sandfly species caught was Sergentomyia minuta, four specimens only. P.pemiciosus and P.mascittii always occurred together and were found to feed - at least occasionally – on human blood. Glucose and fructose were identified in wild-caught sandflies of both species. A breeding place of P.mascittii was identified in the basement of a house in the middle of Sessa. Analyses of soil samples revealed moderately loamy humus with a slightly acidic pH (5.6).     

Species composition and seasonal abundance of sandflies (Diptera: Psychodidae: Phlebotominae) in coffee agroecosystems

The composition and seasonal occurrence of sandflies were investigated in coffee agroecosystems in the Soconusco region of Chiapas, Mexico. Insect sampling was performed on three plantations located at different altitudes: Finca Guadalupe Zajú [1,000 m above sea level (a.s.l.)], Finca Argovia (613 m a.s.l.) and Teotihuacán del Valle (429 m a.s.l.). Sandflies were sampled monthly from August 2007-July 2008 using three sampling methods: Shannon traps, CDC miniature light traps and Disney traps. Sampling was conducted for 3 h during three consecutive nights, beginning at sunset. A total of 4,387 sandflies were collected during the course of the study: 2,718 individuals in Finca Guadalupe Zajú, 605 in Finca Argovia and 1,064 in Teotihuacán del Valle. The Shannon traps captured 94.3% of the total sandflies, while the CDC light traps and Disney traps captured 4.9% and 0.8%, respectively. More females than males were collected at all sites. While the number of sandflies captured was positively correlated with temperature and relative humidity, a negative correlation was observed between sandfly numbers and rainfall. Five species of sandflies were captured: Lutzomyia cruciata , Lutzomyia texana , Lutzomyia ovallesi , Lutzomyia cratifer / undulata and Brumptomyia sp. Lu. cruciata , constituting 98.8% of the total, was the most abundant species. None of the captured sandflies was infected with Leishmania spp.     

Phlebotomine sandfly responses to carbon dioxide and human odour in the field

Responses of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) to carbon dioxide (CO2) and human odour were investigated by field experiments in Parana State, southern Brazil. Catches of two predominant species: Lu. intermedia (Antunes & Coutinho) and Lu. whitmani Lutz & Neiva, were compared between traps baited with a human adult or with CO2 emitted at the human-equivalent rate. When the baits were only 40 cm apart, no difference of attractiveness was detected. When baits were separated by 20 m, however, significantly fewer sandflies (44% Lu. intermedia, 46% Lu. whitmani) were trapped with CO2 compared with human bait. This is the first field evidence that anthropophilic sandflies are attracted by human kairomones in addition to CO2. For both species [Lutzomyia intermedia and Lu. whitmani] [corrected], the proportion of human attractiveness attributable to CO2 was significantly more [corrected] for males than females; for Lu. intermedia males human bait was no more attractive than CO2 alone. Gender differences in sandfly olfactory sensitivity are likely to be associated with behavioural differences on the host, where females feed on blood and males find mates. With traps 20 m apart, both Lutzomyia spp. showed roughly linear increased responses (log-log scale) to 0.08-0.55% CO2 equivalent to 0.5-4 humans. This would explain why host size is generally proportional to attractiveness, as observed for other species of phlebotomine sandflies.     

Identification of bloodmeals in Anopheles quadrimaculatus and Anopheles punctipennis from eastern equine encephalitis virus foci in northeastern U.S.A.

The host‐feeding patterns of Anopheles quadrimaculatus Say and Anopheles punctipennis (Say) were examined in order to evaluate their potential contributions to the transmission of eastern equine encephalitis virus (EEEv) and other arboviruses in the northeastern U.S.A. Engorged mosquitoes of the two species were collected from EEEv foci in central New York (NY) and throughout New Jersey (NJ), and their bloodmeals were identified using a polymerase chain reaction (PCR)‐based assay and sequencing portions of the mitochondrial cytochrome b gene. Analysis of 131 An. quadrimaculatus and 107 An. punctipennis from NY revealed that 97.7% and 97.2%, respectively, had acquired blood solely from mammalian hosts. Similarly, examination of 288 An. quadrimaculatus and 127 An. punctipennis from NJ showed 100% and 96.0%, respectively, contained mammalian‐derived bloodmeals. Mosquitoes containing mixed bloodmeals from both avian and mammalian hosts were detected in 1.6% of An. quadrimaculatus from NY, and 2.8% and 4.0% of An. punctipennis from NY and NJ, respectively. White‐tailed deer (Odocoileus virginianus) constituted the most common vertebrate host for these anopheline mosquitoes, accounting for 85.8–97.7% of all bloodmeals identified. The predominance of white‐tailed deer as a source of bloodmeals supports enzootic amplification of deer‐associated arboviruses in this region, including Jamestown Canyon, Cache Valley and Potosi viruses. One horse‐ and two human‐derived bloodmeals were also detected in An. quadrimaculatus collected in NJ. Limited avian‐derived bloodmeals were detected from mourning dove (Zenaida macroura), sharp‐shinned hawk (Accipiter striatus) and house finch (Carpodacus mexicanus), mostly in mixed bloodmeals. Occasional feeding on avian hosts suggests that these mosquitoes may participate as epizootic–epidemic bridge vectors of EEEv from viraemic birds to mammalian hosts of concern, including horses and humans. An isolate of EEEv was recovered from the head and thorax of an An. punctipennis mosquito collected in NY.     

Entomological and parasitological study on phlebotomine sandflies in central and northern Albania

An entomological survey was carried out in two districts of central (Kruje) and northern (Lezhe) Albania. Six collecting sites, showing a variety of diurnal resting sites, were monitored for adult sandflies from June through October 2002. Flies were collected with CDC miniature light traps, sticky traps and mechanical or hand aspirators in peridomestic sites, in bedrooms and inside cow barns, chicken coops and pigpens. All collecting sites monitored were found positive for sandflies. A total of 849 specimens were caught (29.2% males) belonging to five Phlebotomus species. Phlebotomus neglectus (75.6%) was the most abundant species followed by P. perfiliewi (14.4%), P. papatasi (4.6%), P. tobbi (3.6%) and P. similis (1.8%). The first adult of P. neglectus appeared on June 11 and the last one was collected on October 16. The highest density for this species was observed at the end of July. A total of 111 blood-fed females were caught from the two areas studied. P. neglectus was the only species found blood fed in Lezhe and the same species was prevalent (56.1%) in Kruje followed by P. perfiliewi (30.3%), P. tobbi (10.6%); P. papatasi was represented by only two specimens. Blood meal origin was determined in 45/66 (68.2%) of the females tested from Kruie district. P. neglectus was found fed on four hosts, showing the following feeding patterns: cow (71.4%), dog (117.1%), chicken (5.7%) and human (5.7%); P. perfiliewi was found fed on cow (80.0%) and chicken (20.0%), P. tobbi on cow (50.0%), chicken (25.0%) and dog (25.0%). One specimen of P. papatasi was found fed on cow. When such prevalences were analysed by the available biomass for each host present at the collecting site, P. neglectus resulted to be an opportunistic feeder rather than exhibiting preferences for any specific animal. PCR analysis of 39 P. neglectus from the Lezhe district gave negative results for the presence of Leishmania DNA.     

Mosquito host-feeding patterns and implications for Japanese encephalitis virus transmission in northern Australia and Papua New Guinea

Japanese encephalitis (JE) virus spread to northern Australia during the 1990s, transmitted by Culex annulirostris Skuse and other mosquitoes (Diptera: Culicidae). To determine the relative importance of various hosts for potential vectors of JE virus, we investigated the host-feeding patterns of mosquitoes in northern Australia and Western Province of Papua New Guinea, with particular attention to pigs, Sus scrofa L. - the main amplifying host of JE virus in South-east Asia. Mosquitoes were collected by CDC light traps baited with dry ice and 1-octen-3-ol, run 16.00-08.00 hours, mostly set away from human habitations, if possible in places frequented by feral pigs. Bloodmeals of 2569 mosquitoes, representing 15 species, were identified by gel diffusion assay. All species had fed mostly on mammals: only <10% of bloodmeals were from birds. The predominant species was Cx. annulirostris (88%), with relatively few (4.4%) bloodmeals obtained from humans. From all 12 locations sampled, the mean proportion of Cx. annulirostris fed on pigs (9.1%) was considerably lower than fed on other animals (90.9%). Highest rates of pig-fed mosquitoes (>30%) were trapped where domestic pigs were kept close to human habitation. From seven of eight locations on the Australian mainland, the majority of Cx. annulirostris had obtained their bloodmeals from marsupials, probably the Agile wallaby Macropus agilis (Gould). Overall proportions of mosquito bloodmeals identified as marsupial were 60% from the Gulf Plains region of Australia, 78% from the Cape York Peninsula and 64% from the Daru area of Papua New Guinea. Thus, despite the abundance of feral pigs in northern Australia, our findings suggest that marsupials divert host-seeking Cx. annulirostris away from pigs. As marsupials are poor JE virus hosts, the prevalence of marsupials may impede the establishment of JE virus in Australia.     

Distribution of phlebotomine sandflies and the rate of their infection with Leishmania promastigotes in the Algarve, Portugal

Four species of sandflies: Phlebotomus (Larroussius) perniciosus Newstead, Sergentomyia minuta (Rondani), Phlebotomus (Paraphlebotomus) sergenti Parrot and Phlebotomus (Larroussius) ariasi Tonnoir, were collected, by aspiration and light traps, from three dog kennels and an area of high prevalence of human and canine visceral leishmaniasis in the Algarve, Portugal. Midgut infections with leishmanial promastigotes were observed only in P. perniciosus with a minimum infection rate of 5%. The engorgement, gravidity, population dynamics and sex ratios of the sandflies, and the temperature, source and altitude of collection sites were examined. Host preferences were noted for P. perniciosus.     

Identification of mosquito bloodmeals using mitochondrial cytochrome oxidase subunit I and cytochrome b gene sequences

Abstract.  Primer pairs were designed and protocols developed to selectively amplify segments of vertebrate mitochondrial cytochrome oxidase subunit 1 (COI) and cytochrome b (Cyt b ) mtDNA from the bloodmeals of mosquitoes (Diptera: Culicidae). The protocols use two pairs of nested COI primers and one pair of Cyt b primers to amplify short segments of DNA. Resultant sequences are then compared with sequences in GenBank, using the BLAST function, for putative host identification. Vertebrate DNA was amplified from 88% of our sample of 162 wild-caught, blood-fed mosquitoes from Oregon, U.S.A. and GenBank BLAST searches putatively identified 98% of the amplified sequences, including one amphibian, seven mammalian and 14 avian species. Criteria and caveats for putative identification of bloodmeals are discussed.     

Sandfly (Diptera: Psychodidae: Phlebotominae) species diversity in an urban area of the municipality of Tapachula,Chiapas, Mexico

Monitoring phlebotomine sandflies in urban areas is key for epidemiological studies in susceptible populations. This paper describes sandfly fauna that were present in an urban area of the municipality of Tapachula, Chiapas, Mexico, and were captured with Shannon and CDC light traps. During February and March of 2014, 1,442 sandflies were captured, specifically Lutzomyia cruciata (Coquillet) (98.8%), Lutzomyia cayennensis cayennensis (Floch and Abonnenc) (0.8%), Lutzomyia chiapanensis (Dampf) (0.3%) and Lutzomyia atulapai (De León) (0.1%). Lu. cruciata was the most abundant and the most frequently trapped species. This is the first record of its remarkable ability to adapt to urban green areas. The three other species trapped represent new records of geographic distribution for the study region. These results indicate the need to establish measures for reducing both human contact with this vector and the risk of possible sites of infection.     

The relationship between CDC light-trap and human-bait catches of endophagic sandflies (Diptera: Psychodidae) in the Peruvian Andes

Abstract. A study was carried out in the Peruvian Andes to test the suitability of CDC light traps for monitoring changes in the human-landing rate of endophagic phlebotomine sandflies, following house-spraying with pyrethroid insecticide. On four pairs of consecutive nights, sandflies were caught inside eight sprayed and eight unsprayed houses, either by human bait or by CDC light traps. The sandflies collected were Lutzomyia verrucarum (97%) and Lu. peruensis (3%), both probable vectors of Leishmania peruviana , and the species composition was unaffected by house-spraying. A non-linear relationship was detected between light-trap and human-bait catches, but the relationship did not diverge significantly from linearity within the range of sandfly abundance found in most houses in the endemic area (i.e. between 3 and 200 sandflies/house-night), and did not differ significantly between sprayed and unsprayed houses. However, light trap catches had a significantly lower proportion of blood-fed females in sprayed than in unsprayed houses, probably due to an insecticidal effect on post-blood-feeding behaviour. The proportion of Lu. verrucarum was significantly higher in light trap than in human bait catches, indicating that Lu. peruensis is either more anthropophilic or less phototropic than Lu. verrucarum.     

Distribution of phlebotomine sandflies, of the subgenus Larroussius, in Morocco

Adult sandflies (Diptera: Psychodidae) of the genus Phlebotomus and of the subgenus Larroussius were collected with sticky traps from thirty-four localities in central and south-western Morocco. Four Larroussius species were present in this area, Phlebotomus perniciosus Newstead, being the most widespread and abundant species particularly in the mountainous area, was sympatric with the closely related species Phlebotomus longicuspis Nitzulescu. Only one male of P. perniciosus displayed the bifid copulatory valves characteristic of the species. All specimens showed copulatory valves curved at their apex. The number of coxite hairs is a taxonomical tool that differentiates this morph of P. perniciosus from P. longicuspis. Phlebotomus ariasi Tonnoir was collected from six mountainous stations. Phlebotomus mariae Rioux et al. was scarce; only five males were collected and were often associated with P. ariasi.     

Identifying the last supper: utility of the DNA barcode library for bloodmeal identification in ticks

Ticks are among the most important vectors of disease in the Northern Hemisphere, and a better understanding of their feeding behaviour and life cycle is critical to the management and control of tick-borne zoonoses. DNA-based tools for the identification of residual bloodmeals in hematophagous arthropods have proven useful in the investigation of patterns of host use in nature. Using a blind test approach, we challenged the utility of the DNA barcode library for the identification of vertebrate bloodmeals in engorged, field-collected Ixodes scapularis. Universal vertebrate primers for the COI barcode region successfully amplified DNA from the host bloodmeal and only rarely amplified tick DNA. Of the 61 field-collected ticks, conclusive genus- and species-level identification was possible for 72% of the specimens. In all but two cases, barcode-based identification of the bloodmeal was consistent with the morphological identification of the vertebrate host the ticks were collected from. Possible explanations for mismatches or ambiguities are presented. This study validates the utility of the DNA barcode library as a valuable and reliable resource for the identification of unknown bloodmeals in arthropod vectors of disease. Future directions aimed at the refinement of these techniques to gain additional information and to improve the amplification success of digested vertebrate DNA in tick bloodmeals are discussed.