Sand Fly Fauna Research Articles

Integrative Approach to Phlebotomus mascittii Grassi, 1908: First Record in Vienna with New Morphological and Molecular Insights.

Sand flies (Diptera: Psychodidae: Phlebotominae) are blood-feeding insects that transmit the protozoan parasites Leishmania spp. and various arthropod-borne (arbo) viruses. While in Mediterranean parts of Europe the sand fly fauna is diverse, in Central European countries including Austria mainly Phlebotomus mascittii is found, an assumed but unproven vector of Leishmania infantum. To update the currently understudied sand fly distribution in Austria, a sand fly survey was performed and other entomological catches were screened for sand flies. Seven new trapping locations of Ph. mascittii are reported including the first record in Vienna, representing also one of the first findings of this species in a city. Morphological identification, supported by fluorescence microscopy, was confirmed by two molecular approaches, including sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) protein profiling. Sand fly occurrence and activity were evaluated based on surveyed locations, habitat requirements and climatic parameters. Moreover, a first comparison of European Ph. mascittii populations was made by two marker genes, cytochrome c oxidase subunit 1 (COI), and cytochrome b (cytb), as well as MALDI-TOF mass spectra. Our study provides new important records of Ph. mascittii in Austria and valuable data for prospective entomological surveys. MALDI-TOF MS protein profiling was shown to be a reliable tool for differentiation between sand fly species. Rising temperatures and globalization demand for regular entomological surveys to monitor changes in species distribution and composition. This is also important with respect to the possible vector competence of Ph. mascittii.

An integrative approach to identify sand fly vectors of leishmaniasis in Ethiopia by morphological and molecular techniques

BackgroundEthiopia is affected by human leishmaniasis caused by several Leishmania species and transmitted by a variety of sand fly vectors of the genus Phlebotomus. The sand fly fauna in Ethiopia is highly diverse and some species are closely related and similar in morphology, resulting in difficulties with species identification that requires deployment of molecular techniques. DNA barcoding entails high costs, requires time and lacks reference sequences for many Ethiopian species. Yet, proper species identification is pivotal for epidemiological surveillance as species differ in their actual involvement in transmission cycles. Recently, protein profiling using MALDI-TOF mass spectrometry has been introduced as a promising technique for sand fly identification.MethodsIn our study, we used an integrative taxonomic approach to identify most of the important sand fly vectors of leishmaniasis in Ethiopia, applying three complementary methods: morphological assessment, sequencing analysis of two genetic markers, and MALDI-TOF MS protein profiling.ResultsAlthough morphological assessment resulted in some inconclusive identifications, both DNA- and protein-based techniques performed well, providing a similar hierarchical clustering pattern for the analyzed species. Both methods generated species-specific sequences or protein patterns for all species except for Phlebotomus pedifer and P. longipes, the two presumed vectors of Leishmania aethiopica, suggesting that they may represent a single species, P. longipes Parrot & Martin. All three approaches also revealed that the collected specimens of Adlerius sp. differ from P. (Adlerius) arabicus, the only species of Adlerius currently reported in Ethiopia, and molecular comparisons indicate that it may represent a yet undescribed new species.ConclusionsOur study uses three complementary taxonomical methods for species identification of taxonomically challenging and yet medically import Ethiopian sand flies. The generated MALDI-TOF MS protein profiles resulted in unambiguous identifications, hence showing suitability of this technique for sand fly species identification. Furthermore, our results contribute to the still inadequate knowledge of the sand fly fauna of Ethiopia, a country severely burdened with human leishmaniasis.Graphical

Characterization of the sand fly fauna in Barbalha, one of the municipalities with the highest leishmaniasis rates in Brazil.

Leishmaniases are a complex of sand fly-borne diseases that are considered a public health issue in several countries. Brazil presents high leishmaniases rates. The South of Ceará State, known as Cariri region, shows worrying statistics mainly on American tegumentary leishmaniasis. In Barbalha, which is one of the municipalities in this region, there is still a lack of studies regarding the local phlebotomine (Diptera: Psychodidae) fauna in order to help clarify the high rates. This study aimed to characterize such fauna by capturing sand flies with light traps during a four-year period. A total of 3730 sand flies were captured, of which 37.8% were females. Fourteen species were found: 13 of the Lutzomyia genus and one of the Brumptomyia genus. Of the Lutzomyia species, four were proven and five had potential involvement in leishmaniasis transmission. Lutzomyia longipalpis was the most common species (66.97%). This predominance, especially in the urban area, indicates its epidemiological importance and adaptation to environmental conditions modified by human activity. In fact, further studies are still required to accurately determine the behavioral features of these vectors in order to guide public health measurements towards its control and prevention.

Detection of Leishmania species (Kinetoplastida, Trypanosomatidae) in phlebotomine sand flies (Diptera, Psychodidae) from Porto Velho, Northern Brazil

Cutaneous Leishmaniasis (CL) is a vector-borne disease caused by Leishmania species and transmitted by infected female sand flies. CL is widely distributed in Brazil, but knowledge about vectors and transmission cycles could be complex according to localities. The sand fly fauna in Rondônia State is extensive, diverse, and largely unexplored. Although the state records a mean of 1,000 CL cases per year, the vectors of CL are unknown. The aim of this study was to assess phlebotomine sand fly fauna composition using diversity indexes (Shannon [H’] and Simpson [1/D]) and to detect the prevalence of Leishmania infection to verify potential vectors in three ecotopes: (i) forest fragment (FF), (ii) forest edge (FE), and (iii) peridomicile (PE). Captures were performed in four rural districts in the municipality of Porto Velho. A total of 7,026 specimens were captured comprising 72 species, and individuals classified in subgenus level. Overall, the most abundant species were Lutzomyia davisi (n: 1,105), Lutzomyia melloi (n: 760), Lutzomyia auraensis (n: 738) and Lutzomyia antunesi (n: 479). Fauna was most diverse in the FF ecotope (H’ = 20.2, 1/D = 11.2), followed by the FE (H’ = 18.0, 1/D = 10.1) and PE (H’ = 16.6, 1/D = 10.1) ecotopes. Leishmania DNA was detected in 24 of 232 pools. In every ecotope, Leishmania naiffi DNA was identified in the following sand fly species: Lu. antunesi, Lu. davisi, Lu. hirsuta hirsuta, Lu. shawi, Lu. sordellii and Lu. (Trichophoromyia) spp. This observation may indicate that a Le. naiffi transmission focus is present in the study localities. In addition, Leishmania lainsoni was detected in Lutzomyia (Trichophoromyia) spp. Our findings show that sand fly fauna in the study localities is diverse, that Leishmania parasites are circulating in all three ecotopes, and that some sand fly species may be implicated in the transmission of Leishmania to humans in localities evaluated.

Sand flies (Diptera: Psychodidae) in eight Balkan countries: historical review and region-wide entomological survey

BackgroundSand flies (Diptera: Psychodidae) are medically important vectors of human and veterinary disease-causing agents. Among these, the genus Leishmania (Kinetoplastida: Trypanosomatidae), and phleboviruses are of utmost importance. Despite such significance, updated information about sand fly fauna is missing for Balkan countries where both sand flies and autochtonous leishmaniases are historically present and recently re-emerging. Therefore, a review of historical data on sand fly species composition and distribution in the region was followed by a large-scale entomological survey in eight Balkan countries to provide a recent update on local sand fly fauna.MethodsThe literature search involved the period 1910–2019. The entomological survey was conducted at 1189 sampling stations in eight countries (Bulgaria, Bosnia and Herzegovina, Croatia, Kosovo, Montenegro, North Macedonia, Serbia and Slovenia), covering 49 settlements and 358 sampling sites between June and October in the years 2014 and 2016, accumulating 130 sampling days. We performed a total of 1189 trapping nights at these stations using two types of traps (light and CO2 attraction traps) in each location. Sampling was performed with a minimal duration of 6 (Montenegro) and a maximal of 47 days (Serbia) between 0–1000 m.a.s.l. Collected sand flies were morphologically identified.ResultsIn total, 8490 sand fly specimens were collected. Morphological identification showed presence of 14 species belonging to genera Phlebotomus and Sergentomyia. Historical data were critically reviewed and updated with our recent findings. Six species were identified in Bosnia and Herzegovina (2 new records), 5 in Montenegro (2 new records), 5 in Croatia (2 new records), 9 in Bulgaria (5 new records), 11 in North Macedonia (1 new record), 10 in Serbia (no new records), 9 in Kosovo (3 new records) and 4 in Slovenia (no new records).ConclusionsThis study presents results of the first integrated sand fly fauna survey of such scale for the Balkan region, providing first data on sand fly populations for four countries in the study area and presenting new species records for six countries and updated species lists for all surveyed countries. Our findings demonstrate presence of proven and suspected vectors of several Leishmania species.Graphical

Study of fauna, activity patterns and Leishmania infection rate of phlebotomine sand flies in Western Iran.

Cutaneous leishmaniasis is a crucial vector-borne disease caused by various species of Leishmania and is transmitted by several species of sandflies. The present study was conducted to describe sand fly fauna on vectors of leishmaniasis and performing molecular identification of Leishmania isolates from them on the Iran-Iraq border. Entomological surveys were done from May to October 2016-2018 in 2 counties (Mehran and Dehloran) of Ilam province, west of Iran. Sandflies were collected by 40 Sticky Traps at each station. Samples were mounted for species identification using morphological characters of the head and abdominal terminalia. DNA was extracted from Phlebotomus papatasi females, and Leishmania isolates were identified through PCR on minicircle kDNA, followed by sequencing. A total of 5592 sandflies including 2 genera of Phlebotomus and Sergentomyia comprising 8 species of sand flies were detected. Leishmania major infection was detected in 3.33% of 300 tested female sandflies. Phlebotomus papatasi was predominant in outdoor and indoor resting places. Phlebotomus papatasi was determined as dominant vector of Leishmania major infection in Mehran and Dehloran counties, West of Iran. It seems the composition of sandfly species in the study area is almost similar to the other parts of Iran. A detailed description of the epidemiology and ecology of Phlebotomine sand flies needs to be established to accomplish effective vector control programs.

Sand fly fauna of Crete and the description of Phlebotomus (Adlerius) creticus n. sp. (Diptera: Psychodidae)

BackgroundThe Greek island of Crete is endemic for both visceral leishmaniasis (VL) and recently increasing cutaneous leishmaniasis (CL). This study summarizes published data on the sand fly fauna of Crete, the results of new sand fly samplings and the description of a new sand fly species.MethodsAll published and recent samplings were carried out using CDC light traps, sticky traps or mouth aspirators. The specific status of Phlebotomus (Adlerius) creticus n. sp., was assessed by morphological analysis, cytochrome b (cytb) sequencing and MALDI-TOF protein profiling.ResultsPublished data revealed the presence of 10 Phlebotomus spp. and 2 Sergentomyia spp. During presented field work, 608 specimens of 8 species of Phlebotomus and one species of Sergentomyia were collected. Both published data and present samplings revealed that the two most common and abundant species were Phlebotomus neglectus, a proven vector of Leishmania infantum causing VL, and Ph. similis, a suspected vector of L. tropica causing CL. In addition, the field surveys revealed the presence of a new species, Ph. (Adlerius) creticus n. sp.ConclusionsThe identification of the newly described species is based on both molecular and morphological criteria, showing distinct characters of the male genitalia that differentiate it from related species of the subgenus Adlerius as well as species-specific sequence of cytb and protein spectra generated by MALDI-TOF mass spectrometry.

Entomological Survey of the Sand Fly Fauna of Kayseri Province: Focus on Visceral and Cutaneous Leishmaniasis in Central Anatolia, Turkey

In Turkey, leishmaniasis occurs in two clinical forms: cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL). CL has been reported mainly from south-eastern Anatolia and the eastern part of the Mediterranean region, whereas VL is sporadic in almost all geographical regions of Turkey. Both clinical forms of the disease have been recorded in Kayseri province for decades, but no study has been conducted on the sand fly fauna in this area. Therefore, we determine the species composition and population dynamics of sand flies prevalent in Kayseri province and identify possible vector species. Data related to the recent locations of CL and VL cases were obtained from the Ministry of Health, and sand flies were collected in 14 localities of different districts using CDC light traps. The specimens were transferred to the laboratory in 70% ethanol, and morphological identification to the species level was performed using previously written keys. A total of 1,636 sand fly specimens were collected. Morphological identification revealed eight species of the genus Phlebotomus (P. perfiliewi s.l., P. halepensis, P. simici, P. major s.l., P. papatasi, P. tobbi, P. sergenti and P. mascittii) and one species of the genus Sergentomyia (S. dentata). Among all specimens, members of subgenus Adlerius (54.58%) formed the dominant group, followed by the subgenus Larroussius (43.76%). Our results showed that P. halepensis, belonging to subgenus Adlerius, and P. perfiliewi, belonging to the subgenus, are probable vectors of cutaneous and VL in the province, respectively.

Vertical stratification of sand fly diversity in relation to natural infections of Leishmania sp. and blood-meal sources in Jamari National Forest, Rond\xf4nia State, Brazil

BackgroundAlmost 1000 cases of American cutaneous leishmaniasis have been registered yearly in Rondônia State, Brazil. Little is known about the Leishmania transmission cycle (vectors and reservoirs) in the state. This study aimed to evaluate sand fly fauna from two vertical stratification layers in order to identify potential vectors and their blood-meal sources.MethodsThe study was conducted in Jamari National Forest. Sand flies were collected in the canopy (15 m) and at ground level (1 m) using HP light traps during four months, February, April, August and October, 2018. Insects were identified to the species level, and females were subjected to DNA extraction and PCR targeting minicircle kDNA and hsp70 (for Leishmania detection and species identification), and cytb (to identify blood-meal sources). Exploratory data analysis was used to determine mean of abundance and species richness between stratifications. The hsp70 and cytb sequences were analyzed and compared with sequences from GenBank.ResultsOverall, 68 species were identified from 15,457 individuals. On the Potosi trail, 7531 individuals of 49 species were collected; canopy captures totaled 6463 individuals of 46 species, while ground captures totaled 1068 individuals of 38 species. On the Santa Maria trail, 7926 individuals of 61 species were collected; canopy captures totaled 6136 individuals of 51 species, while ground captures totaled 1790 individuals of 53 species. A total of 23 pools were positive for kDNA (canopy n = 21, ground n = 2). Only two samples were sequenced for hsp70 (both in canopy); one sequence exhibited similarity with Leishmania braziliensis (Lutzomyia davisi pool) and another with L. naiffi (Lu. antunesi pool). The cytb fragment was amplified in 11 of 86 samples. Sample sequencing identified cytb DNA from 5 blood-meal sources: Micrastur gilvicollis, Psophia viridis, Tamandua tetradactyla, Homo sapiens and Choloepus didactylus.ConclusionsSand fly fauna is more diverse in the canopy than at ground level. Factors such as blood-meal sources, resting sites, and abiotic components probably contribute to high abundance in the canopy. Our results reinforce the possibility that Lu. antunesi and Lu. davisi participate in Leishmania transmission in forest environments and may play an important role in transmission from sylvatic to human hosts.

Molecular detection of Leishmania (Sauroleishmania) tarentolae in human blood and Leishmania (Leishmania) infantum in Sergentomyia minuta: unexpected host-parasite contacts.

The detection of atypical Kinetoplastida in vertebrate hosts and vectors might suggest unexpected host-parasite contacts. Aside to major vectors of Leishmania (Leishmania) infantum in Italy (e.g. Phlebotomus perniciosus and Phlebotomus perfiliewi), the sand fly fauna also includes Sergentomyia minuta, herpetophilic and proven vector of Leishmania (Sauroleishmania) tarentolae, in which records of blood meal on mammals and detection of L. infantum DNA are increasing. This study was conducted in Central Italy aiming to molecularly detect potential atypical Leishmania host-vector contacts. Detection of Leishmania spp. DNA was performed by polymerase chain reaction (SSU rRNA, ITS1 targets) on field-collected sand fly females (N=344), blood samples from humans (N=185) and dogs (N=125). Blood meal identification was also performed on engorged sand flies. Leishmania spp. DNA was found in 13.1% sand flies, 3.7% humans and 14.4% dogs. Sequence analysis identified L. infantum in S. minuta (4.4%), P. perniciosus (9.1%), humans (2.2%) and dogs (14.4%). Leishmania tarentolae was detected in S. minuta (12.6%), P. perfiliewi (6.6%) and human (1.6%) samples. Of 28 S. minuta examined for blood meal, 3.6 and 21.4% scored positive for human and lizard DNA, respectively. These results indicate the importance of one-health approach to explore new potential routes of transmission of leishmaniasis involving S. minuta.

Entomological Investigations, Seasonal Fluctuations and Impact of Bioclimate Factors of Phlebotomines Sand Flies (Diptera: Psychodidae) of an Emerging Focus of Cutaneous Leishmaniasis in Aichoun, Central Morocco.

Leishmaniasis diseases are endemic in Morocco. An entomological survey was conducted in Aichoun locality for 1 year from September 2013 to August 2014. The objective of this study was to investigate the sand fly fauna, mainly the species composition and the monthly species prevalence in accordance with bioclimate factors. Sand flies were collected twice a month, using sticky traps and CDC light traps. During a one-year study, 4472 specimens of sand fly were caught (72.56% male/22.44% female) that were composed of seven species divided into two genera: Phlebotomus (99.46%) and Sergentomyia (0.53%). Ph. sergenti was the most prevalent species (46.64%), followed by Ph. perniciosus (38.19%), Ph. longicuspis (9.32%), Ph. papatasi (5.23%), and Ph. ariasi (0.06%). The genus Sergentomyia was even less frequent. The population dynamics showed a bimodal trend with two peaks: the first one in October (12.03% of specimens) and the second in June (27.92% of specimens). The study of the effects of climatic factors in the study area showed a link between the dynamics of sand flies and the variation of these parameters (temperature and relative humidity). During the period between November and March, the sand flies were absent. The highest prevalence of sand flies was recorded in June when the temperature and relative humidity values reached, respectively, 25.8°C and 42%. The minimum number of specimens was collected in September with an average temperature of 23.19°C and relative humidity of 57.4%. Statistical analysis with principal component analysis has shown a strong positive correlation between temperature parameters and the seasonal distribution of sand flies. The climatic factor of relative humidity has been judged of being negatively correlated. The wind speed does not have any impact on the relative abundance of all species. Within this context, the results will be useful for the development of a monitoring program to better manage the operations and evaluate their effectiveness.

Diversity of phlebotomine sand flies and molecular detection of trypanosomatids in Brumadinho, Minas Gerais, Brazil.

This study aimed to describe the sand fly fauna and detect trypanosomatids in these insects from Casa Branca, state of Minas Gerais, Brazil, an endemic area of both visceral (VL) and tegumentary leishmaniasis (TL). Sand flies were collected bimonthly from May 2013 to July 2014, using automatic light traps exposed for three consecutive nights in peridomiciliary areas of nine houses with previous reports of VL and TL. ITS1-PCR and DNA sequencing were performed for trypanosomatids identification. A total of 16,771 sand flies were collected belonging to 23 species. The most abundant species was Nyssomyia whitmani (Antunes & Coutinho, 1939) (70.9%), followed by Lutzomyia longipalpis (Lutz & Neiva, 1912) (15.2%) and Migonemyia migonei (França, 1920) (9.1%). Leishmania amazonensis DNA was detected in Ny. whitmani (four pools) and Le. braziliensis DNA was detected in Psychodopygus lloydi (one pool). In seven pools of Ny. whitmani and in one pool of Lu. longipalpis positive for Leishmania DNA, the parasite species was not determined due to the low quality of the sequences. Moreover, DNA of Herpetomonas spp. was detected in Ny. whitmani (two pools) and Cortelezzii complex (one pool). DNA of Crithidia spp. was detected in Ny. whitmani and Ps. lloydi (both one pool). Our results suggest that Ny. whitmani may be involved in the transmission of Le. amazonensis in the study area. The molecular detection of Le. amazonensis suggests the presence of this species in a sylvatic cycle between vertebrate and invertebrate hosts in the region of Casa Branca. Our data also reveal the occurrence of other non-Leishmania trypanosomatids in sand flies in Casa Branca District.

Diurnal adult resting sites and breeding habitats of phlebotomine sand flies in cutaneous leishmaniasis endemic areas of Kurunegala District, Sri Lanka

BackgroundSand flies are responsible for the transmission of several disease pathogens including Leishmania. Sand flies breed in habitats with high levels of humidity and organic matter. They are nocturnal in nature and peak activity ranges from dusk to dawn. The scientific evidence on breeding ecology and diurnal resting sites of sand fly fauna are important aspects of planning and implementing vector control activities. However, such fundamental information is grossly inadequate in Sri Lanka to support the control efforts in the country. Therefore, the present study addresses some of the important aspects of sand fly breeding ecology and diurnal resting sites.MethodsPotential resting sites were thoroughly observed, and sand flies were collected using a battery-operated aspirator and sticky papers when appropriate from three selected Medical Officer of Health (MOH) areas (Polpithigama, Maho and Galgamuwa) in Kurunegala district, Sri Lanka. Soil samples were collected from each potential breeding site. Half of each soil sample was incubated for 45 days. The other half was screened for immature stages. Adult sand flies collected from field and emerged adults at the insectary under confined incubation were identified using morphological characteristics.ResultsPepper bushes and termite mounds were the most notable resting sites while, betel bushes, cattle huts, piles of coconut shells, latrines, manna bushes and tree holes were also positive for sand fly adults. Only two species, Phlebotomus argentipes and Sergentomyia punjabensis, were reported. Soil samples were collected from a total of 432 sites and 7 of them were positive for immature stages. Predominant breeding habitats identified during the present study were mud flats and moist soils of rice paddies, the soil below decaying hay, drying irrigational tank bottom moist soil, and the floors of cattle huts.ConclusionThis study demonstrates that the potential adult resting sites and breeding habitats are abundant in the Polpithigama, Maho and Galgamuwa MOH areas. Therefore, vector control activities targeting both adult and immature stages of sand flies are recommended.

Entomological Survey for the Detection of Sand Fly Fauna and Vector Species in the Cutaneous Leishmaniasis Endemic Area in East Mediterranean Region of Turkey, Mersin Province.

Cutaneous (CL) and visceral (VL) forms of leishmaniasis, transmitted by sand flies, are seen in all countries located in Mediterranean Basin including Turkey. In this study, we aimed to conduct an entomological survey for the detection of sand fly fauna and vector species in Mersin province, one of the important endemic areas for CL in Turkey. In total, 912 sand fly specimens were collected in 2010 and 2011 using CDC light traps. Nine Phlebotomus (Diptera: Psychodidae) and three Sergentomyia (Diptera: Psychodidae) species were detected. Of the collected Phlebotomus sand flies, P. sergenti Parrot, 1917 (30.1%) was the most dominant followed by P. alexandri Sinton, 1928 (18.2%), P. neglectus/syriacus Tonnoir Adler (12.0%), P. tobbi Adler & Theodor, 1930 (11.7%), and P. papatasi Scopoli, 1786 (10.2%), while S. minuta Rondani, 1843 (11.3%) was the dominant species among Sergentomyia. During the field work in 2011, female specimens (n = 81) were screened for the presence of Leishmania promastigotes by midgut dissection, and all were found negative. The rest of the collected female specimens (n = 334) were pooled according to species (P. alexandri, P. neglectus/syriacus, P. papatasi, P. sergenti, P. simici, and P. tobbi) and location (Mut, Silifke, and Anamur). In total, 29 pools were generated and real-time ITS1 PCR assay was performed to detect and identify natural Leishmania Ross, 1903 (Kinetoplastida: Trypanosomatida) infection. Two pools, both from Mut town, containing P. sergenti specimens were found positive and Leishmania tropica Ross, 1903 was identified as an infectious agent for both pools. In conclusion, the sand fly fauna was determined in an endemic area for CL. The detection of L. tropica DNA in P. sergenti specimens showed the possible vectorial role of this species in Mersin province.

Sandfly surveillance and investigation of Leishmania spp. DNA in sandflies in Kosovo.

In the past decade, leishmaniasis seems to be re-emerging in Balkan countries. There are serious implications that Kosovo is a visceral leishmaniasis endemic region with autochthonous transmission; nevertheless, surveillance of vectors, reservoirs or the disease is not yet established. Gaining knowledge about sandfly vector species is a prerequisite for the development of a monitoring and control plan in the future. After a long gap in research of over 70 years, sandfly studies in Kosovo were resumed in 2014. During this presence/absence study, nine sandfly species were detected: Phlebotomus papatasi, Ph. perfiliewi, Ph. tobbi, Ph. neglectus, Ph. simici, Ph. balcanicus, Ph. alexandri, Ph. mascittii and Sergentomyia minuta. Three species are new with regard to the fauna of Kosovo-Ph. alexandri, Ph. balcanicus and Ph. mascittii. Besides increased diversity, changes in the number of collected specimens and distribution range of species were recorded, with Ph. neglectus being the most dominant species with the widest distribution. Testing of randomly chosen females for Leishmania spp. DNA resulted the in detection of L. tropica in a specimen of Ph. neglectus. The presence of numerous vector species in the sandfly fauna of Kosovo pose a threat for the re-emergence of vector-borne diseases. Therefore, continuous surveillance is recommended with regular updates on vector distribution and abundance.

Lutzomyia evandroi in a New Area of Occurrence of Leishmaniasis.

Sand flies are vectors of medical and veterinary concern, responsible for the transmission of Leishmania parasites. These invertebrates are widely distributed throughout the world and their abundance and diversity is dependent on several biotic and physical factors. Based on the epidemiological importance of these insects, the aim of this study was to assess the sand fly fauna of a new area of occurrence of leishmaniasis. From October 2018 to September 2019, samplings were performed using CDC light traps at six different collection points located in a high altitude area in the municipality of Garanhuns, Northeastern Brazil. Geographic coordinates were recorded using the satellite remote sensing Global Positioning System (GPS) Garmin Etrex20. The spatial distribution of sand flies was evaluated using kernel density estimation (KDE). Engorged females (n = 12) were molecularly processed to search for Leishmania DNA. A total of 138 sand fly specimens were collected during the study period, with 100% (138/138) identified as Lutzomyia evandroi. Climatic conditions did not influence the number of sand flies (males and females) collected monthly, but a statistically significant difference was observed between the number of specimens and different collections points. The kernel map showed a heterogeneous spatial distribution of sand flies in the municipality of Garanhuns with a hotspot in the south of the region. Data presented herein are pivotal in filling the gap on the epidemiology of leishmaniasis in this high altitude area, serving as an alert to local health authorities. It also indicated that the putative role of L. evandroi in the transmission of Leishmania parasites should be clarified.

Ecology of Sand Flies (Psychodidae: Phlebotominae) and Natural Infection of Pintomyia townsendi With Leishmania amazonensis in a Cutaneous Leishmaniasis Focus in Colombia.

Humans have influenced the epidemiological patterns of American cutaneous leishmaniasis by habitat disturbance, which has led to the emergence of new transmission foci. In these transmission areas, detecting natural infection of sand fly species with Leishmania parasites is of prime importance in epidemiological studies. In this study, we examined the species composition, spatial distribution, seasonality, and natural infection with Leishmania of the sand fly fauna in an emergent leishmaniasis focus located in Colombia. Sand flies were collected from September 2014 to June 2015 using CDC light traps located in indoor, peridomestic, and outdoor habitats within areas with confirmed clinical patients. In total, 13,488 sand flies of 12 species and seven genera were collected. Among these, Pintomyia townsendi (Ortiz) was the most abundant species comprising 76.3% of total flies collected. The sand fly richness and abundance were influenced by habitat and weather conditions. Outdoor areas harbored the greatest diversity of sand flies. Rainfall negatively affected abundance, whereas increased temperature showed a low positive effect. We detected Leishmania amazonensis parasites in Pi. townsendi for the first time.

Species composition of sand flies (Diptera: Psychodidae) in caves of Quadrilátero Ferrífero, state of Minas Gerais, Brazil.

Caves are extreme and inhospitable environments that can harbor several species of vertebrate and invertebrate animals. Among these animals are phlebotomine sand flies, vectors of parasites of the genus Leishmania that cause leishmaniasis. This study aimed to evaluate the species composition of sand flies of four caves, a cave located at Moeda Sul (MS) and three at Parque Estadual Serra do Rola Moça (PESRM), in the region of the Quadrilátero Ferrífero in the state of Minas Gerais, southeastern Brazil. Sand flies were collected with automatic light traps. Non-Metric Multidimensional Scaling, using a dissimilarity matrix calculated with the Jaccard index, and Multivariate Permutation Analysis were used to evaluate sand fly species composition among entrance, interior, and the surrounding environments of each sampled cave and to infer biological mechanisms from patterns of distribution among these different cave environments. A total of 375 phlebotomine sand flies representing 14 species and six genera were collected. The most abundant species were Evandromyia tupynambai (54.7%), Brumptomyia troglodytes (25.6%), Evandromyia edwardsi (6.1%), Psathyromyia brasiliensis (4.8%) and Lutzomyia longipalpis (4.3%). Thirty individuals were collected at MS, 16 inside the cave and 14 from its surroundings. At PESRM, five individuals were collected from the surroundings of cave RM38, 190 individuals from cave RM39 (48 in the cave and 142 from its surroundings) and 150 individuals from cave RM40 (42 in the cave and 108 from its surroundings). The results revealed a rich sand fly fauna with similar species compositions among the entrance, interior, and surrounding environments of each sampled cave, suggesting that both caves and their surroundings are important for maintaining sand fly communities.

Potential Vectors of Leishmania Parasites in a Recent Focus of Visceral Leishmaniasis in Neighborhoods of Porto Alegre, State of Rio Grande do Sul, Brazil.

In view of recent cases of human and canine visceral leishmaniasis reported in Porto Alegre, Rio Grande do Sul, Brazil, we investigated the sand fly fauna inhabiting the neighborhoods of Morro Santana and Jardim Carvalho, Brazil, continuing a series of entomological surveys aimed to identify potential vectors of Leishmania (Ross, 1903) parasites. Sand flies were collected monthly from October 2016 to October 2017 using CDC light traps in the intradomiciliary and peridomiciliary environments of seven residences. Sand fly abundance was correlated to climatic variables. Females were pooled by species, location, and date for Leishmania DNA molecular screening using ITS1 and kDNA polymerase chain reaction. In total, 501 sand flies from five species were collected in which Lutzomyia gaminarai (Cordero, Vogelsang & Cossio, 1928) (Diptera: Psychodidae) (78%) was the most abundant species in the intradomiciliary sites while Migonemyia migonei (Franca, 1920) (Diptera: Psychodidae) (43.3%) was the most abundant in the peridomiciliary sites. A higher number of sand flies were collected during the warmest months, from December to March (Mann-Whitney statistical test - P < 0.001). Leishmania infantum DNA was detected in Lu. gaminarai (2), Pintomyia fischeri (Pinto, 1926) (1) and Mg. migonei (1). Leishmania braziliensis DNA was detected in Lu. gaminarai (1) and Pi. fischeri (1). Our results add support to the possible vector role of Pi. fischeri in the epidemiological cycle of Le. infantum in Brazil. Furthermore, the first documented detection of Leishmania DNA in Lu. gaminarai may be indicative of multiple vectors being involved in the Leishmania cycle within Porto Alegre.

Sandfly fauna and behavior (Diptera: Psychodidae) in municipalities of the Mesoregion North Pioneer of Paraná, Brazil

The occurrence of cutaneous leishmaniasis (CL) in the municipalities of Jaboti, Japira, Pinhalão, and Tomazina in the North Pioneer of the State of Paraná, where this disease is endemic, prompted the investigation of sandfly fauna, their population changes throughout the year, and their behavior in and around human accommodation. In these municipalities, the collection of sandflies was conducted using Falcão traps from 7 pm to 6 am once a month. 32,994 sandflies, 18,442 males and 14,552 females, were collected and represented by the species Brumptomyia brumpti, Brumptomyia cunhai, Evandromyia correalimai, Evandromyia cortelezzii, Expapillata firmatoi, Micropygomyia ferreirana, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, Pintomyia fischeri, Pintomyia monticola, Pintomyia pessoai, and Psathyromyia bigeniculata. Ny. neivai (SISA = 0.9808) predominated in Japira (46.9%), Pinhalão (53.1%), and Tomazina (38.9%), while Ny. whitmani (SISA = 0.9423) predominated in Jaboti (53.8%). Most sandflies were captured in domestic animal shelters, with a peak in September. The high number of sandflies collected in domestic animal shelters, species richness, and presence of species involved in the epidemiology of CL reveal the need for permanent entomological surveillance in the municipalities mentioned. The use of phlebotomine control measures in these municipalities should be considered in conjunction with the anthropogenic actions that have resulted in changes to sandfly fauna and behavior.