scholarly journals Development of a Diagnostic Marker for Phlebotomus papatasi to Initiate a Potential Vector Surveillance Program in North America

Insects ◽  
2018 ◽  
Vol 9 (4) ◽  
pp. 162 ◽  
Author(s):  
Austin Merchant ◽  
Tian Yu ◽  
Jizhe Shi ◽  
Xuguo Zhou
Keyword(s):  
Rapid Identification ◽  
International Travel ◽  
Sand Fly ◽  
Surveillance Program ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Diagnostic Assay ◽  
Vector Surveillance ◽  
Geographical Regions ◽  
Imminent Threat

Phlebotomus papatasi, an Old World sand fly species, is primarily responsible for the transmission of leishmaniasis, a highly infectious and potentially lethal disease. International travel, especially military rotations, between domestic locations and P. papatasi-prevalent regions in the Middle East poses an imminent threat to the public health of US citizens. Because of its small size and cryptic morphology, identification of P. papatasi is challenging and labor-intensive. Here, we developed a ribosomal DNA-polymerase chain reaction (PCR)-based diagnostic assay that is capable of detecting P. papatasi genomic DNA from mixed samples containing multiple sand flies native to the Americas. Serial dilution of P. papatasi samples demonstrated that this diagnostic assay could detect one P. papatasi from up to 255 non-target sand flies. Due to its simplicity, sensitivity and specificity, this rapid identification tool is suited for a long-term surveillance program to screen for the presence of P. papatasi in the continental United States and to reveal geographical regions potentially vulnerable to sand fly-borne diseases.

Detection of Pintomyia fischeri (Diptera: Psychodidae) With Leishmania infantum (Trypanosomatida: Trypanosomatidae) Promastigotes in a Focus of Visceral Leishmaniasis in Brazil

2020 ◽  
Author(s):  
Fredy Galvis-Ovallos ◽  
Adriele Eiko Ueta ◽  
Gabriella de Oliveira Marques ◽  
Ana Maria Casagrande Sarmento ◽  
Gabriela Araujo ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
Species Complex ◽  
Natural Infection ◽  
Metropolitan Region ◽  
Sand Fly ◽  
Surveillance Program ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Vector Surveillance

Abstract Visceral leishmaniasis is spreading in Brazil where the main vector of its agent, Leishmania infantum Nicolle, 1908, is the Lutzomyia longipalpis (Lutz & Neiva, 1912) species complex (Diptera: Psychodidae: Phlebotominae), on which many of the activities of the visceral leishmaniasis surveillance program are based. However, there are areas where canine, and/or human cases have been occurring without the presence of this species complex as in the western part of the Greater São Paulo Metropolitan region, where Embu das Artes municipality is situated. In this area, Pintomyia fischeri (Pinto, 1926) has been implicated as potential vector of Le. infantum but so far its natural infection with this parasite has not yet been ascertained. Therefore, the present study sought to investigate the natural infection in sand flies of a CVL focus in Embu das Artes. The sand fly collections were undertaken with Shannon and CDC traps, monthly, between 1800 and 2100 hours from November 2018 to October 2019, inclusive. A total of 951 sand flies (457 males and 494 females), belonging to 10 species, were captured. Pintomyia fischeri was the predominant species (89.5%); of which 426 females were dissected and one of them (0.23%) was found to be harboring flagellates in its midgut. A sample of these flagellates was isolated in culture and characterized by a 234 base pair fragment of Leishmania heat-shock protein 70 gene (hsp70) and restriction fragment length polymorphism with Hae III restriction enzyme as Le. infantum. This finding reinforces previous evidence of Pi. fischeri as a vector of Le. infantum in foci of visceral leishmaniasis and highlights the importance of vector surveillance in areas where this species occurs.

scholarly journals CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Isabelle Louradour ◽  
Kashinath Ghosh ◽  
Ehud Inbar ◽  
David L. Sacks
Keyword(s):  
Immune Deficiency ◽  
Immune Response ◽  
Leishmania Major ◽  
Vector Competence ◽  
Targeted Mutagenesis ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Content Type ◽  
Imd Pathway

ABSTRACT Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report, we adapted CRISPR (clustered regularly interspaced palindromic repeat)/Cas9 (CRISPR-associated protein 9) technology to the Phlebotomus papatasi sand fly, a natural vector for Leishmania major, targeting the sand fly immune deficiency (IMD) pathway in order to decipher its contribution to vector competence. We established a protocol for transformation in P. papatasi and were able to generate transmissible null mutant alleles for Relish (Rel), the only transcription factor of the IMD pathway. While the maintenance of a homozygous mutant stock was severely compromised, we were able to establish in an early generation their greater susceptibility to infection with L. major. Flies carrying different heterozygous mutant alleles variably displayed a more permissive phenotype, presenting higher loads of parasites or greater numbers of infective-stage promastigotes. Together, our data show (i) the successful adaptation of the CRISPR/Cas9 technology to sand flies and (ii) the impact of the sand fly immune response on vector competence for Leishmania parasites. IMPORTANCE Sand flies are the natural vectors of Leishmania parasites. Studies of sand fly/Leishmania interactions have been limited by the lack of successful genomic manipulation of these insects. This paper shows the first example of successful targeted mutagenesis in sand flies via adaptation of the CRISPR/Cas9 editing technique. We generated transmissible null mutant alleles of relish, a gene known to be essential for the control of immune response in other insects. In addition to the expected higher level of susceptibility to bacteria, the mutant flies presented higher loads of parasites when infected with L. major, showing that the sand fly immune response impacts its vector competence for this pathogen.

scholarly journals Diversity and Geographical Distribution of Sand Flies Phlebotomus papatasi (Diptera: Phlebotominae) by using Geometric Morphometric Technique from two Iraqi Provinces

2020 ◽  
Vol 17 (3) ◽  
pp. 0754
Author(s):  
Sahar Abd Abd ◽  
Riyadh Okail ◽  
Soolaf A. Kathiar ◽  
Nawras Mzahem
Keyword(s):  
Geographical Distribution ◽  
Morphological Features ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Centroid Size ◽  
Geometric Morphometric ◽  
Geometric Morphometry ◽  
Size And Shape

            The variation in wing morphological features was investigated using geometric morphometric technique of the Sand Fly from two Iraqi provinces Babylon and Diyala . We distributed eleven landmarks on the wings of Sand Fly species. By using the centroid size and shape together, all species were clearly distinguished.  It is clear from these results that the wing analysis is an essential method for future geometric morphometry studies to distinguish the species of Sand Flies in Iraq.

scholarly journals Seasonal dynamics of Phlebotomus Papatasi (Scopoli, 1786) (Diptera: Psychodidae) population in southern Republic of Moldova

2021 ◽  
Author(s):  
Tatiana Sulesco ◽  
Keyword(s):  
Seasonal Dynamics ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Light Traps ◽  
Phlebotomine Sand Flies ◽  
Republic Of Moldova ◽  
Genus Leishmania

Phlebotomine sand flies are vectors of several infectious pathogens, including parasitic protozoans of the genus Leishmania and phleboviruses. Increasing sand fly biting nuisance reported by residents from southern Republic of Moldova since 2011 initiated this study. Ceadir-Lunga, a semi-urban locality in southern Republic of Moldova was selected for seasonal sand fly collections outdoors and indoors in 2015 and 2017 using CDC light traps and manual aspirators. Continuous trapping showed markedly longer activity of P. papatasi indoors. Specimens were collected from first aspirations in the second half of June until last collections in mid-September, suggesting that the actual indoor activity of P. papatasi may have been longer. Low numbers of trapped specimens do not allow make accurate conclusions regarding the seasonal dynamics.

scholarly journals Diversity and Dynamics of Sand Flies (Diptera: Psychodidae) of Two Cutaneous Leishmaniasis Foci in the Fes-Boulemane Region of Northern Morocco

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Fatima Zahra Talbi ◽  
Chafika Faraj ◽  
Fouad EL-Akhal ◽  
Fatiha El Khayyat ◽  
Driss Chenfour ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Statistical Tests ◽  
Control Measures ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Phlebotomus Perniciosus ◽  
Phlebotomus Sergenti ◽  
Sergentomyia Minuta ◽  
Two Peaks

Cutaneous leishmaniasis (CL) is an infectious disease caused by various species ofLeishmaniaand transmitted by several species of sand flies (Diptera: Psychodidae). In order to evaluate the risk of leishmaniasis transmission in Fes-Boulemane, an investigation was carried out in two localities, Aichoun and Bouasseme, during 2011. From January to December, 1120 specimens were collected in Aichoun comprising six species belonging to two genera:Phlebotomus sergenti(76.07%),Phlebotomus longicuspis(9.01%),Phlebotomus perniciosus(8.48%),Phlebotomus papatasi(4.82%),Sergentomyia minuta,andSergentomyia fallax. For Bouasseme, seven species were identified withPhlebotomus sergenti(60.39%) dominating, followed byPhlebotomus perniciosus(20%) andPhlebotomus longicuspis(12.15%). The remaining species,Phlebotomus papatasi,Phlebotomus ariasi,Sergentomyia minuta,andSergentomyia fallax,were less prevalent. The activity of sand flies in both localities is marked by the dominance ofPh. sergentiwith two peaks occurring in June and September. In order to obtain a better understanding of sand fly diversity among their species, results were analyzed by the ecological indices determinant: specific richness, the relative abundance, and Shannon-Weiner index (H′). Further studies of sand fly diversity should employ statistical tests and molecular analyses. This study can be useful in the implementation of appropriate future control measures.

scholarly journals Comparison of sand fly trapping approaches for vector surveillance of Leishmania and Bartonella species in ecologically distinct, endemic regions of Peru

2021 ◽  
Vol 15 (7) ◽  
pp. e0009517
Author(s):  
Victor O. Zorrilla ◽  
Marisa E. Lozano ◽  
Liz J. Espada ◽  
Michael Kosoy ◽  
Clifton McKee ◽  
...  
Keyword(s):  
Citrate Synthase ◽  
Light Trap ◽  
Amazon Region ◽  
Sand Fly ◽  
Sand Flies ◽  
Vector Surveillance ◽  
Bartonella Bacilliformis ◽  
Blue Led ◽  
Red Led ◽  
Bg Sentinel

Background In Peru, the information regarding sand fly vectors of leishmaniasis and bartonellosis in the Amazon region is limited. In this study, we carried out sand fly collections in Peruvian lowland and highland jungle areas using different trap type configurations and screened them for Leishmania and Bartonella DNA. Methodology/Principal findings Phlebotomine sand flies were collected in Peruvian Amazon jungle and inter Andean regions using CDC light trap, UV and color LED traps, Mosquito Magnet trap, BG Sentinel trap, and a Shannon trap placed outside the houses. Leishmania spp. screening was performed by kDNA PCR and confirmed by a nested cytochrome B gene (cytB) PCR. Bartonella spp. screening was performed by ITS PCR and confirmed by citrate synthase gene (gltA). The PCR amplicons were sequenced to identify Leishmania and Bartonella species. UV and Blue LED traps collected the highest average number of sand flies per hour in low jungle; UV, Mosquito Magnet and Shannon traps in high jungle; and Mosquito Magnet in inter Andean region. Leishmania guyanensis in Lutzomyia carrerai carrerai and L. naiffi in Lu. hirsuta hirsuta were identified based on cytB sequencing. Bartonella spp. related to Bartonella bacilliformis in Lu. whitmani, Lu. nevesi, Lu. hirsuta hirsuta and Lu. sherlocki, and a Bartonella sp. related to Candidatus B. rondoniensis in Lu. nevesi and Lu. maranonensis were identified based on gltA gene sequencing. Conclusions/Significance UV, Blue LED, Mosquito Magnet and Shannon traps were more efficient than the BG-Sentinel, Green, and Red LED traps. This is the first report of L. naiffi and of two genotypes of Bartonella spp. related to B. bacilliformis and Candidatus B. rondoniensis infecting sand fly species from the Amazon region in Peru.

scholarly journals Designing and Introducing a New Artificial Feeding Apparatus for Sand Fly Rearing

2019 ◽  
Author(s):  
Mahboubeh Fatemi ◽  
Zahra Saeidi ◽  
Parviz Noruzian ◽  
Amir Ahmad Akhavan
Keyword(s):  
Disease Transmission ◽  
Artificial Feeding ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Feeding Apparatus ◽  
Medical Sciences ◽  
Membrane Feeding ◽  
Chicken Skin ◽  
New Apparatus

Background: Due to strict ethical rules, the risk of accidental disease transmission and the most importantly, inconven­ience regarding using of live animals, artificial feeding apparatus has been developed for colonization of haematophagous insects. Rearing of sandfly is more difficult than other haematophagous insects. Methods: In the current study, a new apparatus for membrane feeding of Phlebotomus papatasi was designed, made and compared with available apparatus in Sand Fly Insectary, Tehran University of Medical Sciences, Tehran, Iran, in 2014. Results: In comparison to other apparatus designed for artificial feeding of other arthropods, our designed apparatus had the highest performance which after up to 1h, the majority of sand flies landed and took blood and among tested membranes, chicken skin was proved the most efficient membrane. Conclusion: Sand fly artificial feeding apparatus can be used at least for rearing of Ph. papatasi.

Larval Conditioning and Aging of Sand Fly Rearing Medium Affect Oviposition Site Selection in Phlebotomus papatasi (Diptera: Psychodidae) Sand Flies

2021 ◽  
Author(s):  
Lindsey R Faw ◽  
Kasie Raymann ◽  
Nayma Romo Bechara ◽  
Gideon Wasserberg
Keyword(s):  
Conditioned Medium ◽  
Food Source ◽  
Food Sources ◽  
Sand Fly ◽  
Larval Rearing ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Larval Food ◽  
Gravid Females ◽  
Larval Conditioning

Abstract Sand fly larvae develop in sheltered humid habitats containing decaying organic matter on which they feed. Previously, we showed that gravid females of Phlebotomus papatasi Scopoli (Diptera: Psychodidae) are attracted to and stimulated to lay eggs on larval rearing medium containing larvae. That study, however, did not control for the possible effect of medium aging. Our goal in this study was to evaluate the effect of larval substrate conditioning on attraction and oviposition responses of Ph. papatasi sand flies while controlling for the effect of substrate aging. Initially, we confirmed that the pretreatment fresh larval food sources (to be used as larval conditioned and unconditioned media) did not differ with respect to their effect on attraction and oviposition responses. The larval conditioned medium was produced by rearing larvae to the second/third-instar stage over 3 wk using the same larval food source. To produce larval unconditioned medium, the same amount of fresh larval food was added to a control rearing cup that did not contain larvae but was aged under identical time and conditions. Two-choice bioassays were conducted to evaluate gravid female’s attraction and oviposition response to larval conditioned and unconditioned media. We found that gravid females were significantly attracted (P < 0.05) to larval conditioned medium when compared with unconditioned medium under the same amount of time and conditions. However, no such difference was found with respect to oviposition response. Both attraction and oviposition responses were significantly increased for larval conditioned and unconditioned media in comparison to the initial fresh larval food source.

scholarly journals Oviposition-Site Selection of Phlebotomus papatasi (Diptera: Psychodidae) Sand Flies: Attraction to Bacterial Isolates From an Attractive Rearing Medium

2020 ◽  
Author(s):  
Madhavi L Kakumanu ◽  
Bahjat F Marayati ◽  
Coby Schal ◽  
Charles S Apperson ◽  
Gideon Wasserberg ◽  
...  
Keyword(s):  
Site Selection ◽  
Bacterial Species ◽  
Oviposition Site ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Bacterial Isolates ◽  
Oviposition Site Selection ◽  
Response Curves

Abstract Phlebotomine sand flies are worldwide vectors of Leishmania parasites as well as other bacterial and viral pathogens. Due to the variable impact of traditional vector control practices, a more ecologically based approach is needed. The goal of this study was to isolate bacteria from the most attractive substrate to gravid Phlebotomus papatasi Scopoli sand flies and determine the role of bacterial volatiles in the oviposition attractancy of P. papatasi using behavioral assays. We hypothesized that gravid sand flies are attracted to bacterially derived semiochemical cues associated with breeding sites. Bacteria were isolated from a larvae-conditioned rearing medium, previously shown to be highly attractive to sand flies. The isolated bacteria were identified by amplifying and sequencing 16S rDNA gene fragments, and 12 distinct bacterial species were selected for two-choice olfactometer bioassays. The mix of 12 bacterial isolates elicited strong attraction at the lower concentration of 107 cells per ml and significant repellence at a high concentration of 109 cells per ml. Three individual isolates (SSI-2, SSI-9, and SSI-11) were particularly attractive at low doses. In general, we observed dose-related effects, with some bacterial isolates stimulating negative and some positive dose–response curves in sand fly attraction. Our study confirms the important role of saprophytic bacteria, gut bacteria, or both, in guiding the oviposition-site selection behavior of sand flies. Identifying the specific attractive semiochemical cues that they produce could lead to development of an attractive lure for surveillance and control of sand flies.

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