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> Living Beings > Eukaryote > Phlebotominae

Phlebotominae

Phlebotominae, also known as sand flies, are a subfamily of small blood-feeding dipteran insects.
They are important vectors for various parasitic diseases, including leishmaniasis, bartonellosis, and certain viral infections.
Phlebotominae species are found worldwide, primarily in tropical and subtropical regions.
Their biology, ecology, and role in disease transmission make them a key focus of research in medical entomology and public health.
PubCompare.ai's AI-driven platform helps optimize this research by identifying the most accurate and reproducible methodolgies across the scientific literature, preprints, and patents.
Streamline your Phlebotominae studies with this powerful tool for improving research quality and efficiency.

Most cited protocols related to «Phlebotominae»

1
Morphological Analysis of Leishmania major
Cited 11 times
Gut smears of L. major-infected females 5, 9 and 12 days PBM were fixed with methanol, stained with Giemsa and examined under the light microscope with an oil-immersion objective. One hundred and sixty randomly selected promastigotes from four sand flies/smears were measured in each combination of Leishmania line and time PBM. Body length, flagellar length and body width of parasites were measured and position of the kinetoplast in relation to the nucleus was examined. Four morphological forms were distinguished, based on the criteria of Walters (1993) (link) and Cihakova and Volf (1997) (link): (i) short promastigotes: body length < 14 µm and flagellar length < 2 times body length; (ii) elongated nectomonads: body length ≥ 14 µm; (iii) metacyclic promastigotes: body length < 14 µm and flagellar length ≥ 2 times body length, and (iv) round forms: body width > 4 µm and body length ≤ 7.5 µm include also paramastigotes with kinetoplast lateral to the nucleus. We use here the term short promastigotes derived from the terminology of Walters (1993) (link) (short nectomonad promastigotes) which is the older synonym of leptomonads (leptomonad promastigotes) proposed by Rogers et al. (2002) (link). Haptomonads cannot be distinguished in this study as they remain attached to the gut and cannot be measured on gut smears.
Sádlová J., Price H.P., Smith B.A., Votýpka J., Volf P, & Smith D.F. (2010). The stage-regulated HASPB and SHERP proteins are essential for differentiation of the protozoan parasite Leishmania major in its sand fly vector, Phlebotomus papatasi. Cellular Microbiology, 12(12), 1765-1779.
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Publication 2010
Cell Nucleus Females Human Body Leishmania Light Microscopy Methanol Parasites Phlebotominae Submersion
2
Leishmania Infection Dynamics in Sand Fly
Cited 8 times
The colony of P. papatasi was maintained at 26°C on 50% sucrose and 14 h light/10 h dark photoperiod as described previously (Benkova and Volf, 2007 (link)). Sand fly females were infected by feeding through a chick-skin membrane on heat-inactivated rabbit blood containing 106 promastigotes ml−1. Engorged sand flies were maintained in the same conditions as the colony and dissected 2, 5, 9 and 12 days PBM. The location of Leishmania infections in the sand fly digestive tract (foregut, SV, thoracic and abdominal midgut, and endoperitrophic and ectoperitrophic space) was determined by dissection and examination by light microscopy. Parasite loads were estimated by two methods: infections seen in the gut in situ were graded according to Myskova et al. (2008 (link)) as light (< 100 parasites per gut), moderate (100–1000 parasites per gut) and heavy (> 1000 parasites per gut). Alternatively, 30–40 guts from females with late infections (10–12 days PBM) were individually dissected into NET 50 and stored in −20°C for qPCR. Sand fly infection experiments were repeated four times for combinations of wild-type (FVI), KO and Kin lines and twice for combinations of FVI, +HASPB and +SHERP lines.
Sádlová J., Price H.P., Smith B.A., Votýpka J., Volf P, & Smith D.F. (2010). The stage-regulated HASPB and SHERP proteins are essential for differentiation of the protozoan parasite Leishmania major in its sand fly vector, Phlebotomus papatasi. Cellular Microbiology, 12(12), 1765-1779.
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Publication 2010
Abdomen Blood Dissection Females Gastrointestinal Tract Infection Intestines Leishmaniasis Light Light Microscopy Parasites Phlebotominae Phlebotomus Rabbits Skin Sucrose Tissue, Membrane Vision
3
Modeling Sand Fly Leishmania Transmission
Cited 7 times
We considered sand flies in the susceptible (SF), latent (EF, not infectious), or infectious (IF) stages. Flies can become infected by blood meals taken on infectious humans (IHP, IHD, IHS, IHT1, IHT2, IHL). The infection rate λF of flies is determined by the following: 1) the biting rate β, 2) the infection probabilities of flies dependent on the infection status of the hosts (pF1 to pF4, see Table 2) and 3) the numbers of infectious hosts.
We assumed that each blood meal of a susceptible sand fly leads to a sand fly infection if taken from a symptomatically infected human (pF3 = 100% for KA patients IHS, IHT1, IHT2 and pF4 = 100% for PKDL patients IHL). For a sensitivity analysis, see Results.
The probability that a fly becomes infected when feeding on asymptomatically infected hosts of type IHP or IHD was estimated with the model as follows. The infection probability pF2 of late asymptomatically infected humans (IHD) was estimated, whereby we assumed that the infectivity of hosts increases monotonically from the time of infection until the late asymptomatic state IHD. We defined an intermediate infection probability for flies originating from hosts in the early asymptomatic state IHP as pF1 = pF2/2.
Stauch A., Sarkar R.R., Picado A., Ostyn B., Sundar S., Rijal S., Boelaert M., Dujardin J.C, & Duerr H.P. (2011). Visceral Leishmaniasis in the Indian Subcontinent: Modelling Epidemiology and Control. PLoS Neglected Tropical Diseases, 5(11), e1405.
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Publication 2011
Asymptomatic Diseases Blood Diptera Homo sapiens Hypersensitivity Infection Patients Phlebotominae Phlebotomus
4
Estimating Canine Leishmania Infectiousness
Cited 6 times
The date of patent infection for dogs and foxes was estimated as the first date at which animals were positive by any serological or parasitological assay; all samples thereafter were considered as infected based on previous analyses demonstrating a very low incidence of serological reversal [31] (link), [35] (link), [36] (link). At each bimonthly examination, dogs were classified according to their total clinical score as asymptomatic (scores 0–2), oligosymptomatic (3–6) and symptomatic (>6). Dogs with >8 months post infection follow-up and all bimonthly clinical scores <3 were considered long-term asymptomatic. Infectiousness was assessed as either positive (≥1 sandfly infected) or negative, or as the proportion of sandflies infected at any single time point (point xenodiagnosis). Dogs were also classified previously [22] (link), [35] (link) as “highly infectious” (>20% of total flies infected), “mildly infectious” (>0% and <20% flies infected), and “non-infectious” (no flies infected) by serial xenodiagnoses (n = 6,002 flies dissected from 173 independent trials): the highly infectious group were shown to be responsible for >80% of all transmission events [22] (link). All foxes were non-infectious (n = 1,469 flies from 44 trials) [35] (link).
Courtenay O., Carson C., Calvo-Bado L., Garcez L.M, & Quinnell R.J. (2014). Heterogeneities in Leishmania infantum Infection: Using Skin Parasite Burdens to Identify Highly Infectious Dogs. PLoS Neglected Tropical Diseases, 8(1), e2583.
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Publication 2014
Animals Biological Assay Canis familiaris Diptera Foxes Infection Phlebotominae Phlebotomus Transmission, Communicable Disease Xenodiagnosis
5
Synthetic Pheromone Trapping of Sand Flies
Cited 6 times
All field experiments aimed to test whether dispensers loaded with synthetic pheromone presented at a test station attracted more females and males than simultaneously presented solvent-only controls. Stations consisted of boxes constructed from four panels of thick (10mm) plywood (55cm (width) by 105cm (height)), held together by plastic garden ties sewn through holes (1.5mm ID) drilled in the corner of each plywood panel. Each panel had three holes (10cm ID) drilled in a horizontal line across the middle to allow host odour to escape. Two opposing panels in each box had a groove across the top edge, onto which a wooden pole could be secured and a trap suspended.
At dusk (17:00-18:00), pairs of experimental stations (test and control) were placed 3m apart in residential gardens, previously identified by the local disease control authority (Centro de Controle de Zoonoses (CCZ)) as sites of sand fly aggregation in or around chicken coops. We began by using lures in conjunction with modified (Natal et al. 1991 ) CDC miniature light traps, recognized as an effective device for sampling sand fly populations. Traps, powered overnight by 6-volt rechargeable batteries, were fitted with 35cm-diameter metal lids, and suspended from the pole above each station. Captured flies were collected in pots (10cm diam × 8cm height) connected to each trap via a nylon sleeve (10cm diam). A chicken, selected at random from the garden, was placed in each box to provide a source of host odor.
A dispenser containing 50μg of synthetic pheromone was attached to the underside of the lid at each test station, and a dispenser containing solvent only was similarly positioned at each control station.
The next morning (07:00-08:00) nylon sleeves were tied with string to prevent collected flies from escaping, before pots and batteries were removed. Chickens were freed from boxes, dispensers removed from lids. The positions of test and control boxes was alternated between nights. Flies were identified as male or female L. longipalpis and counted in the laboratory under x40 magnification, and trap batteries recharged before reuse. This entire first trial was repeated a second time, with the light bulbs, which are attractive when lit, removed.
Under natural conditions, male sand flies only aggregate on or near host animals, and host odor has been shown to synergize female attraction to sex pheromone in the laboratory (Bray and Hamilton 2007 (link)). However, females can also be attracted to pheromone alone in the laboratory. To test the attractiveness of synthetic pheromone without host odor in the field, the first two experiments were repeated without chickens in test stations.
Finally, to test whether pheromone could be used to trap flies in conjunction with a cheaper alternative to mechanical light traps, the first trial (with chickens) was repeated, replacing CDCs with blue, 28cm wide agricultural sticky traps (Russell IPM, Deeside, UK) wrapped around the top of each box. Each morning, sticky traps were numbered and covered with clear film, before being removed to the laboratory for counting.
Each experiment was conducted over two to four nights, using six to seven pairs of boxes spread across two to three gardens. New dispensers were used for each night of experiments. Wilcoxon signed rank tests (Sokal and Rohlf 1995 ) compared numbers of flies captured at test and control boxes in each pair. Males and females were analyzed separately, and pairs excluded from analysis if at least one of the traps did not function correctly.
Bray D.P., Bandi K.K., Brazil R.P., Oliveira A.G, & Hamilton J.G. (2009). Synthetic Sex Pheromone Attracts the Leishmaniasis Vector Lutzomyia longipalpis (Diptera: Psychodidae) to Traps in the Field. Journal of medical entomology, 46(3), 428-434.
Publication 2009
Animals ARID1A protein, human chenodeoxycholate sulfate conjugate Chickens Diptera Females Light Males Marijuana Abuse Medical Devices Metals Nylons Odors Pheromone Phlebotominae Phlebotomus Plant Bulb Population Group Sex Attractants Solvents Zoonoses

Most recents protocols related to «Phlebotominae»

1
Molecular Detection of Canine Leishmaniasis
Genomic DNA was extracted from the EDTA-blood samples, conjunctival swabs and sand flies using the DNeasy Blood & Tissue Kit (Qiagen GmbH, Hilden, Germany), according to the manufacturer’s instructions.
Blood samples, conjunctival swabs and sand fly DNA samples were tested for L. infantum DNA using a TaqMan real-time quantitative PCR (qPCR) assay that targeted a 120-bp fragment of the kinetoplast minicircle DNA, as previously reported [13 (link)]. A positive (reference DNA sample from blood, conjunctival swab and sand fly DNA, respectively) and a negative (PCR grade water) control were included in each qPCR run.
The qPCR was performed only on samples collected on SDs 0 and 630 and, in addition, on DNA blood samples at all time points from: (i) all dogs that tested positive at the beginning of the study and became negative during the study; (ii) all dogs that tested positive by ELISA only; and (iii) all dogs that tested only as low positive on qPCRs performed on swab DNA samples on SD 630.
For E. canis, Anaplasma platys and A. phagocytophilum, specific PCR tests were performed on all DNA from blood samples on SD 0 and SD 630 collected from dogs with at least 1 year follow-up. To determine the presence of E. canis, a PCR targeting a 345-bp fragment of the 16S ribosomal RNA (rRNA) gene [14 (link)] of various species, including E. canis, Ehrlichia chaffeensis, Ehrlichia muris, Ehrlichia ruminantium, A. phagocytophilum, A. platys, Anaplasma marginale, Anaplasma centrale, Wolbachia pipentis, Neorickettsia sennetsu, Neorickettsia risticii and Neorickettsia helminthoeca, was performed according to previously described thermal-cycling conditions [15 (link)].
To identify the presence of Anaplasma DNA, we performed species-specific nested PCRs for A. platys and A. phagocytophilum DNA targeting a 678-bp and a 546-bp fragment of the 16S rRNA gene, respectively, according to previously described protocols [16 (link)].
A positive (E. canis and A. platys or A. phagocytophilum reference DNA sample) and a negative (PCR grade water) control were included in each PCR run. Amplification products were visualised on 1.5% agarose gels stained with ethidium bromide.
PCR products (from samples positive for E. canis and A. platys/phagocytophilum) were sent to a commercial service (CeMIA SA, Larissa, Greece) for purification and sequencing on both strands (Sanger sequencing). The results were assembled with Seqman 8.1 software (DNASTAR, Madison, WI, USA). Assembled sequences were aligned using the Basic Local Alignment Tool (BLAST) and compared with reference sequences using the MegAlign application of the Lasergene software package (DNASTAR).
Ligda P., Gizzarelli M., Kostopoulou D., Foglia Manzillo V., Saratsis A., Saratsi K., Michler S., Ringeisen H., Boegel A., Schunack B., Pollmeier M., Kontrafouris M., Tsatsaki O., Oliva G, & Sotiraki S. (2023). Determination of the effect of collars containing 10% w/w imidacloprid and 4.5% w/w flumethrin (Seresto®) on the incidence of Leishmania and other canine vector-borne pathogen infections in Greece. Parasites & Vectors, 16, 89.
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Publication 2023
Anaplasma Anaplasma centrale Anaplasma marginale Anaplasma platys Biological Assay BLOOD Canis familiaris Conjunctiva DNA, A-Form DNA, Kinetoplast Edetic Acid Ehrlichia chaffeensis Ehrlichia muris Ehrlichia ruminantium Enzyme-Linked Immunosorbent Assay Ethidium Bromide Gels Genome Neorickettsia helminthoeca Neorickettsia risticii Neorickettsia sennetsu Nested Polymerase Chain Reaction Phlebotominae Real-Time Polymerase Chain Reaction Ribosomal RNA Genes Sepharose Tissues Wolbachia
2
Sand Fly Sampling and Identification
Sand flies were collected starting on SD 0 in both locations. Follow-up collections in Heraklion (Crete) were performed between SDs 78 and 80, SDs 134 and 136, SDs 318 and 320, SDs 409 and 411, SDs 533 and SD 534 and SD 661 and 663. Follow-up collections in Leros were performed between SDs 77 and 80, SDs 144 and 146, SDs 338 and 340, SDs 424 and 426, SDs 539 and 542 and SDs 675 and 678. The US Center for Disease Control (CDC) miniature light traps and mechanical aspirators were used for all collections. The light traps were placed close to animal-inhabited biotopes. The traps operated overnight (for 2–4 consecutive nights each time), and the sand flies collected were kept either dry or in 70% ethanol until examination.
All captured sand flies were counted. For species identification, the head and the terminal part of the abdomen of all collected female sand flies were mounted on permanent microscopy slides and subsequently identified using the appropriate keys based on the morphology of the pharynx and the genitalia armature and documented [11 , 12 ]. Upon species identification, female sand flies of the same species and collected at the same time point from the same trap were homogenised in groups of five and kept frozen at − 20 °C until further molecular analysis.
Ligda P., Gizzarelli M., Kostopoulou D., Foglia Manzillo V., Saratsis A., Saratsi K., Michler S., Ringeisen H., Boegel A., Schunack B., Pollmeier M., Kontrafouris M., Tsatsaki O., Oliva G, & Sotiraki S. (2023). Determination of the effect of collars containing 10% w/w imidacloprid and 4.5% w/w flumethrin (Seresto®) on the incidence of Leishmania and other canine vector-borne pathogen infections in Greece. Parasites & Vectors, 16, 89.
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Publication 2023
Abdomen Animals Ethanol Freezing Genitalia Head Light Microscopy Pharynx Phlebotominae Woman
3
Socioeconomic Factors Impact on Vector-Borne Diseases
The total number of features used for socioeconomic data is 13. Most features are regarding the water, i.e., people have access to pure water or how they treat water before drinking, etc. Stagnant and dirty water is the primary cause of malaria. Dirty puddles and dirty water are high mosquito breeding places and are some of the reasons for malaria [41 (link)]. The reasons for poverty are less availability of health facilities and the environmental conditions of people’ s living and working areas. Some specific groups of the population are at risk for leishmaniasis. In rural and peri-urban areas, houses are built with poor-quality materials (mud, straw, bamboo), which can become the reason for sandflies’ shelter and can cause leishmaniasis. A lack of sewage and garbage collection services, leading to the accumulation of waste, can also become the reason for vector breeding places [42 (link)]. Socioeconomic factors have an impact also on dengue. The dengue epidemic remains due to socioeconomic factors [43 (link)].
Javaid M., Sarfraz M.S., Aftab M.U., Zaman Q.U., Rauf H.T, & Alnowibet K.A. (2023). WebGIS-Based Real-Time Surveillance and Response System for Vector-Borne Infectious Diseases. International Journal of Environmental Research and Public Health, 20(4), 3740.
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Publication 2023
Cloning Vectors Culicidae Dengue Fever Epidemics Garbage Leishmaniasis Malaria Phlebotominae Population Group Sewage
4
Culturing Leishmania Promastigotes for Experimentation
Cited 1 time
L. (L.) amazonensis and L. (L.) infantum promastigotes were cultured in 199 medium (Sigma Aldrich, Burlington, MA, USA) supplemented with 0.005% hemin, 40 mM HEPES pH 7.4, 100 µM adenine, 4 mM sodium bicarbonate, 20 μg/mL gentamicin, and 10% FBS. Promastigotes of LV79 (MPRO/BR/72/M1841, obtained from the rodent Proechimys sp. from Pará State, Brazil) and PH8 (IFLA/BR/67/PH8, isolated from the sand fly Lutzomyia flaviscutellata from Pará State, Brazil) strains were obtained through cultivation of amastigotes derived from BALB/c mice lesions in medium 199 at 24 °C. Parasites were subcultured weekly to an initial density of 2 × 106 parasites/mL until the eighth passage. For that, BALB/c mice infected with LV79 and with PH8 were kept in our animal facility. Amastigotes were collected from the footpad lesions when promastigotes’ cultures reached 6–7 passages.
All experiments employed parasites at fourth day of culture (early stationary phase); macrophage infections employed day 4 and day 6 cultures.
L. (L.) infantum Ba262 strain (MCAN/BR/1989/BA262) was used as control for LPG assays.
Tano F.T., Telleria E.L., Rêgo F.D., Coelho F.S., de Rezende E., Soares R.P., Traub-Cseko Y.M, & Stolf B.S. (2023). Phenotypical Differences between Leishmania (Leishmania) amazonensis PH8 and LV79 Strains May Impact Survival in Mammal Host and in Phlebotomine Sand Flies. Pathogens, 12(2), 173.
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Publication 2023
Adenine Animals Bicarbonate, Sodium Biological Assay Gentamicin Hemin HEPES Infection Lutzomyia Macrophage Mice, Inbred BALB C Parasites Phlebotominae Rodent Strains
5
Artificial Sandfly Infection Protocol
For artificial infection of sandflies, defibrinated rabbit blood was used. Red blood cells were centrifuged and washed in sterile PBS 3 times and serum was inactivated at 56 °C for 30 to 45 min. The parasites were collected on early log phase culture, resuspended in the inactivated serum, and later mixed with the red blood cells. Artificial infections were performed with blood containing 107 parasites/mL of blood using a Hemotek artificial feeder.
Tano F.T., Telleria E.L., Rêgo F.D., Coelho F.S., de Rezende E., Soares R.P., Traub-Cseko Y.M, & Stolf B.S. (2023). Phenotypical Differences between Leishmania (Leishmania) amazonensis PH8 and LV79 Strains May Impact Survival in Mammal Host and in Phlebotomine Sand Flies. Pathogens, 12(2), 173.
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Publication 2023
Blood Erythrocytes Infection Parasites Phlebotominae Rabbits Serum Sterility, Reproductive

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More about "Phlebotominae"

Phlebotominae, also known as sand flies, are a subfamily of small blood-feeding dipteran insects that are important vectors for various parasitic diseases, including leishmaniasis, bartonellosis, and certain viral infections.
These insects are found worldwide, primarily in tropical and subtropical regions.
Their biology, ecology, and role in disease transmission make them a key focus of research in medical entomology and public health.
To optimize Phlebotominae research, scientists often utilize tools like the DNeasy Blood & Tissue Kit or the QIAmp DNA Mini Kit for DNA extraction, and the IQ SYBR Green Supermix or IQ Real-Time PCR Systems for quantitative PCR analysis.
The High Pure PCR Template Preparation Kit and DNAzol can also be used for DNA isolation from tissue samples.
Additionally, cell culture media like FBS (Fetal Bovine Serum) may be used to maintain sand fly colonies in the lab.
Researchers may also employ imaging equipment, such as the D70 camera, to capture high-quality images and videos of these insects.
PubCompare.ai's AI-driven platform helps streamline Phlebotominae research by identifying the most accurate and reproducible methodologies across the scientific literature, preprints, and patents.
This powerful tool can improve research quality and efficiency, ensuring that scientists have access to the best protocols and products for their Phlebotominae studies.