First record of a rare case of accidental urinary myiasis by Telmatoscopus albipunctata (Diptera: Psychodidae), with morphological description, in a human in Brazil.

Telmatoscopus albipunctata is an insect of the Psychodidae family, with a worldwide distribution, particularly in tropical and subtropical countries. Although it does not have hematophagous habits, it has veterinary medical importance due to the mechanical transmission of protozoa and bacteria, many of which cause nosocomial infections. This dipteran has been reported as one of the causative agents of accidental myiasis in humans in several countries, and since it has not been registered in South America, the objective of the present report was to describe a rare case of accidental urinary myiasis caused by T. albipunctata (Diptera: Psychodidae) in humans in Brazil. In the present report, a 25-year-old female patient, resident of the central region of the state of Rio Grande do Sul, Brazil, sought medical attention after finding larvae in her urine. She complained of vaginal itching and skin dermatitis. The larvae were sent to the Veterinary Parasitology Laboratory of the Federal University of Santa Maria, where they were mounted on permanent slides for further taxonomic identification. Morphological characteristics allowed the identification of fourth-stage larvae and pupae of T. albipunctata. Therefore, the present work reports the first record of accidental urinary myiasis caused by T. albipunctata infection in Brazil and South America.

A Review of Historical Phlebotominae Taxonomy (Diptera: Psychodidae).

Psychodidae is a group constituted by ca. 3400 species distributed in six existing and one extinct subfamily. Among them, Phlebotominae is of relevant medical and veterinary interest as vectors of pathogens, such as viruses, bacteria, and trypanosomatides, to vertebrates. The taxonomy of Phlebotominae, begun in 1786, received a great boost at the beginning of the twentieth century, when some of its species began to be implicated as vectors of leishmaniasis agents. Currently, the group consists of 1060 species/subspecies described for both hemispheres. Its taxonomy and systematics have been largely based on the morphological characters of adults in view of the limited number of species known in their immature forms, as well as by molecular approaches. This review focuses on the evolution of the phlebotomine systematics, with information regarding when the descriptions of the sand fly species/subspecies occurred, where their type localities are, the number of authors involved in their descriptions, and the researchers who have most contributed to the description of these taxa and their respective institutions. The morphological characters of adult forms used in the taxonomy of groups, in an evolutionary approach, and the current state of knowledge based on immature forms is also presented.

Is Psathyromyia shannoni (Diptera: Psychodidae: Phlebotominae) a species complex? Retrospective study of genetic diversity of COI gene, pathogens and geographic distribution.

The sand fly Psathyromyia shannoni is a broadly distributed species that is relevant for the transmission of pathogens such as Leishmania, Bartonella and viruses in several countries of America. This species belongs to the Shannoni complex. Yet its identification is difficult due to morphologic intraspecific polymorphisms that make it difficult to distinguish between species, and could therefore lead to misidentification and overestimation of its distribution. The aim of this study was to perform a retrospective study on the genetic diversity of Pa. shannoni based on the Cytochrome Oxidase subunit 1 gene and considering its geographic distribution to achieve a better identification and differentiation from other species of the Shannoni complex. According to the Maximum Likelihood analysis and the data on the genetic structure, we propose a modified delimitation of Pa. shannoni species by classifying it into at least three genetic lineages, based on genetic variability and distribution. However, more genetic information on the COI gene, mainly from countries where this species has been reported, is needed to strengthen this proposal.

Morphological description of Pintomyia (Pifanomyia) veintemillasi n. sp., a new sand fly species from the sub-Andean region of Bolivia.

BACKGROUND: Most sand fly species are located in the Americas; some act as vectors of leishmaniasis and other human diseases. In Bolivia, about 25% of Neotropical species have been identified, and only a few have been implicated as vectors of cutaneous and visceral leishmaniasis. A new species of anthropophilic sand fly from the sub-Andean region of Alto Beni is described herein. METHODS: A large systematic entomological survey was carried out in a subtropical humid forest located in the Marimonos mountain range, at around 900 m altitude, in the municipality of Palos Blancos, Sud Yungas Province, Department of La Paz, Bolivia. Sand flies were captured over a period of 26 months between January 1982 and February 1984, at the ground and canopy level, using both CDC light traps and protected human bait. A total of 24,730 sand flies were collected on the ground, distributed in 16 species, and 3259 in the canopy, with eight species. One of these species was labeled as Pintomia (Pifanomyia) nevesi, although certain morphological features allowed us to doubt that it was that taxon. To define the identity of this sand fly, a re-evaluation (this work) was recently carried out through morphological analyses and measurements of the available specimens mounted on Euparal, previously labeled as Pi. (Pif.) nevesi. RESULTS: Based on the morphological traits and measurements, the re-evaluated specimens were definitively identified as a new sand fly species, Pintomyia (Pifanomyia) veintemillasi, closely related to Pi. (Pif.) nevesi and Pintomyia (Pifanomyia) maranonensis within the Evansi series. This new sand fly was the third most numerous anthropophilic species at the floor (6.2%) and the second most numerous anthropophilic at the canopy (35.1%). CONCLUSIONS: A new anthropophilic sand fly species is described as Pi. (Pif.) veintemillasi n. sp. This sand fly species was caught at about 900 m altitude in the Marimonos mountain range, a highly endemic area for cutaneous and mucosal leishmaniasis due to Leishmania (Viannia) braziliensis. Therefore, this species could be involved in the leishmaniasis transmission in the sub-Andean foothills of Alto Beni, Department of La Paz, Bolivia.

Significance of eggshell morphology as an additional tool to distinguish species of sand flies (Diptera: Psychodidae: Phlebotominae).

Morphological characteristics of eggshells are important in sand fly ootaxonomy. In this study, eggshells from Phlebotomus stantoni Newstead, Sergentomyia khawi (Raynal), and Grassomyia indica (Theodor) sand flies collected in Chiang Mai province, Thailand were examined and characterized using light microscopy (LM) and scanning electron microscopy (SEM). Then, eggshell morphology of these three species was described for the first time. Each gravid female was forced to lay eggs by decapitation and the eggs were collected for SEM analysis. Egg laying females were identified by morphological examination and molecular typing using cytochrome b (Cytb) as a molecular marker. The chorionic sculpturing of Ph. stantoni eggs combines two patterns on the same egg: unconnected parallel ridges and reticular patterns. Sergentomyia khawi and Gr. indica have similar chorionic polygonal patterns, but their exochorionic morphology and aeropylar area are different. Results indicate that eggshell morphological characteristics such as chorionic pattern, exochorionic morphology, inter-ridge/boundary area, aeropylar area (including the number of aeropyles) and basal layer, can be useful to develop morphological identification keys of eggs. These can serve as an additional tool to distinguish species of sand flies. In addition, the chorionic sculpturing of the eggs of the three species of sand flies observed by LM is useful for species identification in gravid females with spermathecae obscured by eggs.

Phlebotomus neglectus (Diptera: Psychodidae): New Insights on Its Presence in Iran Based on Three Independent Genetic Loci.

The idea of the existence of Phlebotomus (Larroussius) neglectus (Diptera: Psychodidae) Tonnoir, 1921 in Iran and the skepticism about the existence of Phlebotomus major s.str. Annandale, 1910 had been grown recently in the country. This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of P. major s.l.Annandale, 1910, specimens collected from different parts of Iran. Two different morphotypes were found among the collected samples based on the shape of the aedeagus, ventrally located hairs of the coxite, and parameral sheets. One morphotype seemed similar to P. neglectus Tonnoir 1921 or P. major krimensis Perfiliv1966 (called here MI.N.K.); the other one was similar to P. neglectus and to some extent to P. notus Artemiev & Neronov 1984 (here called MII.N.NO). Cytochrome B, elongation factor 1-alpha, and internal transcribed spacer II loci were amplified, sequenced, and characterized. High sequence homology (98-100%) was observed between P. neglectus and these morphotypes, and phylogenetic analysis was also concordant. Phlebotomus neglectus sequences available in GenBank are located as the sister group of sequences here, particularly near to morphotype MII.N.NO. Moreover, ITS2 locus provides the maximum resolution for differentiation of two morphotypes. Based on achieving results, although a strong support for the presence of P. neglectus was provided, but it is too early to say that P. major s.str. does/does not exist in Iran. This question could be resolved by studying more samples and, most importantly, by comparing the topotypes of P. neglectus and P. major s. str. if possible in the future.

Examination of the interior of sand fly (Diptera: Psychodidae) abdomen reveals novel cuticular structures involved in pheromone release: Discovering the manifold.

The males of many species of New World Phlebotomines produce volatile terpenoid chemicals, shown in Lutzomyia longipalpis s.l. to be sex/aggregation pheromones. Pheromone is produced by secretory cells which surround a cuticular reservoir which collects the pheromone and passes it through a cuticular duct to the surface of the insect. The pheromone then passes through specialised cuticular structures on the abdominal surface prior to evaporation. The shape and distribution of the specialised structures are highly diverse and differ according to species. In this study we used SEM to examine the interior cuticular pheromone collection and transport structures of 3 members of the Lu. longipalpis s.l. species complex and Migonemyia migonei. We found a new structure which we have called the manifold which appears to be a substantial extension of the interior tergal cuticle connected in-line with the cuticular duct and reservoir. The manifold of the Campo Grande member of the complex is longer and wider than the Jacobina member whereas the manifold of the Sobral member was shorter than both other members of the complex. Overall, the secretory apparatus of the Sobral member was smaller than the other two. The manifold of M. migonei was very different to those found in Lu. longipalpis s.l. and was positioned in a pit-like structure within the tergal cuticle. The secretory reservoir was connected by a short duct to the manifold. Differences in the size and shape of the manifold may be related to the chemical structure of the pheromone and may have taxonomic value. Examination of the interior cuticle by SEM may help to locate the secretory apparatus of vector species where pheromonal activity has been inferred from behavioural studies but the external secretory structures or pheromones have not yet been found.

Micro-CT visualization of a promastigote secretory gel (PSG) and parasite plug in the digestive tract of the sand fly Lutzomyia longipalpis infected with Leishmania mexicana.

Leishmaniasis is a debilitating disease of the tropics, subtropics and southern Europe caused by Leishmania parasites that are transmitted during blood feeding by phlebotomine sand flies (Diptera: Psychodidae). Using non-invasive micro-computed tomography, we were able to visualize the impact of the laboratory model infection of Lutzomyia longipalpis with Leishmania mexicana and its response to a second blood meal. For the first time we were able to show in 3D the plug of promastigote secretory gel (PSG) and parasites in the distended midgut of whole infected sand flies and measure its volume in relation to that of the midgut. We were also able to measure the degree of opening of the stomodeal valve and demonstrate the extension of the PSG and parasites into the pharynx. Although our pilot study could only examine a few flies, it supports the hypothesis that a second, non-infected, blood meal enhances parasite transmission as we showed that the thoracic PSG-parasite plug in infected flies after a second blood meal was, on average, more than twice the volume of the plug in infected flies that did not have a second blood meal.

Redescription of Two Psathyromyia Species (Diptera: Psychodidae), Including Description of the Female of Psathyromyia pradobarrientosi Using Molecular and Morphological Approaches.

The taxonomic identity of two species of sand flies, Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004) and Psathyromyia runoides (Fairchild & Hertig, 1953) (Diptera, Psychodidae), was evaluated morphologically and molecularly based upon specimens collected in Brazilian states. The morphological component compared collected specimens with paratypes of Pa. runoides and Pa. pradobarrientosi and their descriptions. Phylogenetic analysis of coI sequences of Pa. pradobarrientosi showed a well-supported group distinct from Pa. runoides. Morphologically, Psathyromyia runoides and Pa. pradobarrientosi males are distinguished by characteristics of the aedeagal ducts and parameral sheath in the genitalia; females are distinguished by the number and shape of the teeth in the cibarium and by the shape of the spermathecae. Given the morphological similarity between the males and the absence of the description of the female of Pa. pradobarrientosi, it is possible that specimens previously identified as Pa. runoides in Brazil are in fact Pa. pradobarrientosi.

TRsandflies: A Web-Based Software for the Morphometric Identification of Sand Flies in Turkey.

Sand flies are vector of several diseases, mostly cutaneous and visceral leishmaniasis (CL and VL). Also, 29 sand fly species have been identified in previous fauna studies carried out in 40 provinces of Turkey. Totally, 24 sand flies species belonging to Phlebotomus (Ph.) (Diptera: Psychodidae) genus have been proven or reported as possible vector species. This study aimed to develop a new software which could contribute to researchers' decision making about the identification of sand flies with obtained data from entomological surveys conducted before in Turkey. Developed software called TRsandflies included 35 textbox created with parameters obtained from caught sand flies specimens by the above-mentioned surveys. It also contained 130 photos and distribution maps related to 24 sand flies species. In addition, C# language and MYSQL database were used in the program. TRsandflies had three different forms (pages) allowing the user to compare the specimens and known species. In the species identification trials with three repetitions carried out in the program, except for the specimens belonging to the Transphlebotomus Artemiev & Neronov, 1984 subgenus, morphometric data of all previously collected sand fly species specimens were included. The process of running the morphometric measurement results of predetermined specimens in the program provided us with an accurate prediction rate of 86.66% in male specimens and 71.66% in female specimens. We concluded that the web-based software developed could play an important role in reducing the rate of possible errors that might be encountered by conventional identification methods.

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

BACKGROUND: The 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. METHODS: All 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. RESULTS: Published 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. CONCLUSIONS: The 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.

Detection of Leishmania infantum DNA in Pintomyia evansi and Lutzomyia longipalpis in Honduras.

BACKGROUND: The two most abundant sand fly species on the Honduran Pacific coast are Lutzomyia (Lutzomyia) longipalpis and Pintomyia (Pifanomyia) evansi. Both species are known vectors of Leishmania (Leishmania) infantum, the etiological agent of visceral leishmaniasis (VL) in the Americas. Although VL and non-ulcerative cutaneous leishmaniasis (NUCL) are endemic on the Pacific versant of the Central American Pacific, the latter is the most frequent manifestation of leishmaniasis there. We evaluated the circulation of Leishmania spp. in the sand fly species on El Tigre Island, an endemic area of NUCL. RESULTS: We collected 222 specimens of six sand fly species. Lu. longipalpis (180 specimens; 81%) and Pif. (Pi.) evansi (35 specimens; 16%) were the most abundant species. L. (L.) infantum DNA was detected in nine of the 96 specimens analyzed; seven of these specimens were identified as Lu. longipalpis, and the remaining two were Pi. evansi, with an infection rate of 9.4% and 2.7%, respectively. CONCLUSION: We present the first record of L. (L.) infantum DNA in Pi. evansi from a NUCL endemic region of Central America. Our results suggest that Pi. evansi could be a secondary vector of L. (L.) infantum in the transmission cycle of leishmaniasis. The detection of natural infections of L. (L.) infantum in sand flies in this region contributes to an understanding of the epidemiology of leishmaniasis in Honduras.

How drain flies manage to almost never get washed away.

Drain flies, Pshycoda spp. (Order Diptera, Family Psychodidae), commonly reside in our homes, annoying us in our bathrooms, kitchens, and laundry rooms. They like to stay near drains where they lay their eggs and feed on microorganisms and liquid carbohydrates found in the slime that builds up over time. Though they generally behave very sedately, they react quite quickly when threatened with water. A squirt from the sink induces them to fly away, seemingly unaffected, and flushing the toilet with flies inside does not necessarily whisk them down. We find that drain flies' remarkable ability to evade such potentially lethal threats does not stem primarily from an evolved behavioral response, but rather from a unique hair covering with a hierarchical roughness. This covering, that has never been previously explored, imparts superhydrophobicity against large droplets and pools and antiwetting properties against micron-sized droplets and condensation. We examine how this hair covering equips them to take advantage of the relevant fluid dynamics and flee water threats in domestic and natural environments including: millimetric-sized droplets, mist, waves, and pools of water. Our findings elucidate drain flies' astounding ability to cope with a wide range of water threats and almost never get washed down the drain.

Morphological identification keys for adults of sand flies (Diptera: Psychodidae) in Sri Lanka.

BACKGROUND: Phlebotomine sand flies are a medically important group of insects that is responsible for the transmission of leishmaniasis. Surveillance plays a major role in vector control programmes through exploring species abundance, potential entomological risk and designing appropriate control measures. In field surveillance programmes of such nature, morphological identification of vector species is of paramount importance. However, in Sri Lanka, there is no published taxonomic key available for the identification of leishmaniasis vectors. METHOD: Identification keys for both male and females of the sand flies recorded in Sri Lanka were developed using morphological features. Main identification features were compared with the original observation of specimens collected from surveys and the use of published literature. Photographic illustrations of morphological features are included with the intention of making the keys user-friendly for non-taxonomists. RESULTS: A total of 22 sand fly species (Diptera: Psychodidae) of the genera Phlebotomus and Sergentomyia reported in Sri Lanka from 1910 to 2019 are included in the present work. CONCLUSION: This simplified key, along with photographs taken from specimens would be beneficial to the health staff, entomologists and research staff who deal with leishmaniasis control programmes and vector-related studies.

Conserved and distinct morphological aspects of the salivary glands of sand fly vectors of leishmaniasis: an anatomical and ultrastructural study.

BACKGROUND: Sand flies are vectors of Leishmania spp., the causative agents of leishmaniasis in vertebrates, including man. The sand fly saliva contains powerful pharmacologically active substances that prevent hemostasis and enhance Leishmania spp. infections. On the other hand, salivary proteins can protect vaccinated mice challenged with parasites. Therefore, sand fly salivary proteins are relevant for the epidemiology of leishmaniasis and can be a potential target for a vaccine against leishmaniasis. Despite this, studies on sand fly salivary glands (SGs) are limited. METHODS: The present study analyzes, in detail, the morphology, anatomy and ultrastructure of the SGs of sand fly vectors of the genera Lutzomyia and Phlebotomus. We used histology, transmission and scanning electron microscopy and lectin labeling associated with confocal laser microscopy. RESULTS: The SGs have conserved and distinct morphological aspects according to the distinct sand fly species. Each SG has a single rounded lobe constituting of c.100-120 secretory cells. The SG secretory cells, according to their ultrastructure and lectin binding, were classified into five different subpopulations, which may differ in secretory pathways. CONCLUSIONS: To the best of our knowledge, these morphological details of sand fly salivary glands are described for the first time. Further studies are necessary to better understand the role of these different cell types and better relate them with the production and secretion of the saliva substances, which has a fundamental role in the interaction of the sand fly vectors with Leishmania.

Morphological and initial molecular characterization of Clogmia albipunctatus larvae (Diptera: Psychodidae) causing urinary myiasis in Egypt.

Myiasis is the infestation of human tissues by dipterous fly larvae of the class Insecta. Clogmia albipunctatus, family Psychodidae, is one of the most medically important insects that cause human myiasis. The aim of the present study is the morphological identification and the molecular characterization of moth flies causing many cases of urinary myiasis in Egypt, based on sequencing of the mitochondrial DNA of the larvae. Seven urinary samples of patients complaining of urinary symptoms and giving a history of low socioeconomic level were examined. Recovered larvae were identified using light microscopy and SEM. For molecular identification, the mitochondrial genes Cytochrome B (cytB), NADH1, NADH1, and 16S were sequenced and phylogenetically analyzed. The morphological and molecular characterization could accurately diagnose our patients to have C. albipunctatus infestation. Such results provided the initial set of data on the molecular identification and phylogenetic analysis of moth flies based on DNA barcoding in Egypt.

On the true identity of Sergentomyia gemmea and description of a closely related species: Se. raynali n. sp.

Several species of Leishmania are responsible for leishmaniases in Thailand, although little is known about their transmission. Sergentomyia gemmea has been suspected several times to transmit Leishmania martiniquensis. Some captures carried out in Thailand and Lao People's Democratic Republic have emphasized the scarcity of Se. gemmea, comprising only 1% of the collected females. The sequencing of cytochrome B mtDNA of our specimens showed that our specimens are not grouped with other Se. gemmea previously deposited in GenBank. The latter are grouped with some Se. khawi and Se. hivernus that we processed in the present study. We suspect misidentifications and propose focusing on the most useful characters for identification of Se. gemmea based on the examination of type-specimens. The examination of the ascoids exhibiting anterior spurs is the most important one. However, we also describe Se. raynali n. sp. exhibiting comparable spurs but differing from Se. gemmea by its original cibarium. Finally, the vectorial role of Se. gemmea appears very questionable in the absence of new evidence.

Traditional and Geometric Morphometry Analyses of Lutzomyia cruciata (Diptera: Psychodidae: Phlebotominae) Populations of Chiapas, Mexico.

The presence of Lutzomyia (Tricholateralis) cruciata (Coquillett 1907) species complex has been suggested by morphological analysis of eggs and genetic studies of females. The present work aimed to compare the diversity in morphology of four populations of Lu. cruciata from the Coast of Chiapas, Mexico, using traditional (TM) and geometric (GM) methods. Several morphological characteristics that were analyzed provided consistency to differentiate at least, three populations of Lu. cruciata. Both methods were effective to detect morphological differences associated with the geographical sites of capture. In both sexes, three and four groups were detected by TM and GM, respectively. These results suggest marked morphological differences in both sexes of Lu. cruciata that make these methods potentially useful to identify the geographical origin of any specimen of this species captured in the study region. Although the results produced by both methods are coincident, geometric morphometrics turned out to be most advantageous with respect to traditional morphometry, since the latter requires more time and effort. The consistency of our results shows that the variability of environmental conditions on the coast of Chiapas determines a high degree of phenotypic plasticity in Lu. cruciata, with the possibility of prezygotic isolation and the formation of species complex.

Three new records of the genus Lutzomyia of the subgenus Helcocyrtomyia (Diptera: Psychodidae: Phlebotominae) from Southwestern Brazilian Amazonia.

The sand fly phlebotomine fauna of the Brazilian municipality of Assis Brasil, Acre was sampled between April 2013 and March 2014 using modified CDC light traps. A total of 22,334 sand flies were collected, 41 of them belonging to three species: Lutzomyia gonzaloi, Lu. kirigetiensis and Lu. munaypata, of the subgenus Helcocyrtomyia, which have previously only been recorded in Peru. This paper presents their first reported collection in Brazil.

Trichophoromyia iorlandobaratai (Diptera: Psychodidae), a new phlebotomine species from the Brazilian Amazonia.

Trichophoromyia iorlandobaratai sp. n. Vasconcelos dos Santos, Santos Neto, Sánchez Uzcátegui & Galardo is described and illustrated on the basis of male specimens collected in the municipality of Itaituba, Pará State, Brazilian Amazonia. This species can be differentiated from other morphologically similar species within Trichophoromyia by means of the male characters such as the ratio between the aedeagal ducts/sperm pump, shape and distribution of setae in the gonocoxite, and particularly the distinct shape of paramere.