Recent Blood Meals Research Articles

Protein mass spectrometry detects multiple bloodmeals for enhanced Chagas disease vector ecology

Chagas disease, a neglected tropical disease endemic in Latin America, is caused by the protozoan parasite Trypanosoma cruzi and is responsible for significant health impacts, especially in rural communities. The parasite is transmitted by insect vectors in the Triatominae subfamily and due to lack of vaccines and limited treatment options, vector control is the main way of controlling the disease. Knowing what vectors are feeding on directly enhances our understanding of the ecology and biology of the different vector species and can potentially aid in engaging communities in active disease control, a concept known as Ecohealth management. We evaluated bloodmeals in rural community, house-caught insect vectors previously evaluated for bloodmeals via DNA analysis as part of a larger collaborative project from three countries in Central America, including Guatemala. In addition to identifying bloodmeals in 100% of all samples using liquid chromatography tandem mass spectrometry (LC-MS/MS) (n = 50), strikingly for 53% of these samples there was no evidence of a recent bloodmeal by DNA-PCR. As individual vectors often feed on multiple sources, we developed an enhanced detection pipeline, and showed the ability to quantify a bloodmeal using stable-isotope-containing synthetic references peptides, a first step in further exploration of species-specific bloodmeal composition. Furthermore, we show that a lower resolution mass spectrometer is sufficient to correctly identify taxa from bloodmeals, an important and strong attribute of our LC-MS/MS-based method, opening the door to using proteomics in countries where Chagas disease is endemic.

Simultaneous identification of host, ectoparasite and pathogen DNA via in-solution capture.

Ectoparasites frequently vector pathogens from often unknown pathogen reservoirs to both human and animal populations. Simultaneous identification of the ectoparasite species, the wildlife host that provided their most recent blood meal(s), and their pathogen load would greatly facilitate the understanding of the complex transmission dynamics of vector-borne diseases. Currently, these identifications are principally performed using multiple polymerase chain reaction (PCR) assays. We developed an assay (EctoBaits) based on in-solution capture paired with high-throughput sequencing to simultaneously identify ectoparasites, host blood meals and pathogens. We validated our in-solution capture results using double-blind PCR assays, morphology and collection data. The EctoBaits assay effectively and efficiently identifies ectoparasites, blood meals, and pathogens in a single capture experiment, allowing for high-resolution taxonomic identification while preserving the DNA sample for future analyses.

Avian malaria parasites in the last supper: identifying encounters between parasites and the invasive Asian mosquito tiger and native mosquito species in Italy.

BackgroundThe invasive Asian tiger mosquito Aedes albopictus has dramatically expanded its distribution range, being catalogued as one of the world’s 100 worst invasive alien species. As vectors of pathogens, Ae. albopictus may create novel epidemiological scenarios in the invaded areas.MethodsHere, the frequency of encounters of Ae. albopictus with the avian malaria parasite Plasmodium and the related Haemoproteus was studied in an area with established populations in northeastern Italy and compared with those from four native mosquito species, Anopheles maculipennis s.l., Culex hortensis, Culex pipiens, and Ochlerotatus caspius. The abdomens of mosquitoes with a recent blood meal were used to identify both the blood meal source and the parasites harboured.ResultsAedes albopictus had a clear antropophilic behaviour while An. maculipennis and Oc. caspius fed mainly on non-human mammals. Birds were the most common hosts of Cx. pipiens and reptiles of Cx. hortensis. Parasites were isolated from three mosquito species, with Cx. pipiens (30%) showing the highest parasite prevalence followed by Cx. hortensis (9%) and Ae. albopictus (5%).ConclusionsThese results are the first identifying the avian malaria parasites harboured by mosquitoes in Italy and represent the first evidence supporting that, although Ae. albopictus could be involved in the transmission of avian malaria parasites, the risk of avian malaria parasite spread by this invasive mosquito in Europe would be minimal.

Crab louse.

A 25-year-old married male presented with itching in the pubic area since two weeks. He gave a history of extramarital contact three weeks ago. On examination he was found to have excoriated papules in lower abdomen, pubic area and thighs. Close inspection revealed multiple brown colored nits attached to the pubic hairs. A clinical diagnosis of phthiriasis pubis was made. Other hair bearing areas including axilla, beard, mustache, eyelids and scalp were examined and were found to be free from infestation. He had no genital ulcer, urethral discharge or inguinal adenopathy. Serology for syphilis and retroviral status were non reactive. Microscopic examination of pubic hair revealed the presence of pubic louse with characteristic morphology and a blood filled esophagus indicating a recent blood meal [Figure 1].Figure 1: Crab louse with its characteristic morphology grasping the hair shaft with blood filled esophagus (40× magnification)The Crab louse or Phthirus pubis is a dorsoventrally flattened, wingless insect of order Anoplura and family Pthiridae. Adult louse measures about 1–2 mm with three parts namely head, thorax with three pairs of legs, and abdomen. The terminal part of each leg has claw-like appendages whose grasp is designed to match the diameter of pubic or axillary hair. The louse buries sharp mouthpiece stylets inside the pubic hair follicle to obtain a constant blood supply.[1] Sexual transmission is considered the most important means of crab louse transmission.[2] However, there are documented cases of transmission from toilet seats, beds, and egg-infested loose hairs dropped by infested persons on shared objects. Treatment options include 1% permethrin rinse, 5% permethrin cream, 1% lindane shampoo, oral ivermectin. Occlusive agents like vaseline, freshly prepared fluorescein eyedrops and manual removal of nits are recommended in case the eyelashes are infested.

Bed bug aggregation pheromone finally identified.

Bed bugs have become a global epidemic and current detection tools are poorly suited for routine surveillance. Despite intense research on bed bug aggregation behavior and the aggregation pheromone, which could be used as a chemical lure, the complete composition of this pheromone has thus far proven elusive. Here, we report that the bed bug aggregation pheromone comprises five volatile components (dimethyl disulfide, dimethyl trisulfide, (E)-2-hexenal, (E)-2-octenal, 2-hexanone), which attract bed bugs to safe shelters, and one less-volatile component (histamine), which causes their arrestment upon contact. In infested premises, a blend of all six components is highly effective at luring bed bugs into traps. The trapping of juvenile and adult bed bugs, with or without recent blood meals, provides strong evidence that this unique pheromone bait could become an effective and inexpensive tool for bed bug detection and potentially their control.

Bed Bug Aggregation Pheromone Finally Identified

AbstractBed bugs have become a global epidemic and current detection tools are poorly suited for routine surveillance. Despite intense research on bed bug aggregation behavior and the aggregation pheromone, which could be used as a chemical lure, the complete composition of this pheromone has thus far proven elusive. Here, we report that the bed bug aggregation pheromone comprises five volatile components (dimethyl disulfide, dimethyl trisulfide, (E)‐2‐hexenal, (E)‐2‐octenal, 2‐hexanone), which attract bed bugs to safe shelters, and one less‐volatile component (histamine), which causes their arrestment upon contact. In infested premises, a blend of all six components is highly effective at luring bed bugs into traps. The trapping of juvenile and adult bed bugs, with or without recent blood meals, provides strong evidence that this unique pheromone bait could become an effective and inexpensive tool for bed bug detection and potentially their control.

Effect of feeding status on mortality response of adult bed bugs (Hemiptera: Cimicidae) to some insecticide products.

Fresh and aged residual deposits of several insecticide products were tested against bed bug adults to determine if a recent bloodmeal affected their mortality response to the residues. The bed bugs with a recent bloodmeal survived significantly longer compared with the unfed ones on their exposure to fresh or aged residual deposits of chlorfenapyr and aged residual deposits of deltamethrin on a wooden substrate. Even though the survival time of fed bed bugs was significantly longer than that of unfed ones on their exposure to fresh residue of deltamethrin and aged residue of desiccant pyrethrin dust, these treatments resulted in similarly high final mortalities regardless of feeding status of the insects. Mortality responses of fed and unfed bed bugs were similar to fresh or aged residual deposits of imidacloprid + cyfluthrin combination and fresh residual deposits of desiccant pyrethrin dust. Topical application assays indicated that a recent bloodmeal significantly increased the bed bug's survival time for chlorfenapyr, but not for deltamethrin. Pyrethroid-resistant bed bugs also showed a similar increase in their survival time for chlorfenapyr after a bloodmeal. The comparison of mortality responses between fed and unfed bed bugs treated with similar amount of chlorfenapyr per fresh body weight indicated that increased body mass was not the primary cause for this bloodmeal-induced tolerance increase for chlorfenapyr. Because the surviving bed bugs can continue ovipositing, the effectiveness of chlorfenapyr residual deposits in bed bug harborages could be significantly affected by the feeding status of the adult bed bug populations.

Effect of blood meal digestion and DNA extraction protocol on the success of blood meal source determination in the malaria vector Anopheles atroparvus

BackgroundHost identification is an essential step in studies on the transmission dynamics of vector-borne disease. Nowadays, molecular tools allow the identification of vertebrate hosts to the species level. However, the proportion of successful identifications is variable and may be affected by the quality of the samples and the laboratory protocols. Here, the effect of two of these factors, namely the digestion status of mosquito blood meal and the DNA extraction procedure, on the success of host identification by amplification and sequencing of a fragment of the cytochrome oxidase 1 gene were tested.MethodsMosquitoes collected both outdoors and indoors during 2012 in southern Spain were identified to species level and their blood meal digestion status recorded using the Sella score, a visual estimation of the digestion status of mosquito blood meals. Each mosquito was assigned randomly to one of two DNA extraction procedures: the quick and cheap HotSHOT procedure or the QIAGEN DNeasy Blood and Tissue® kit and their hosts identified by a molecular method.ResultsThree hundred and forty-seven blood-fed mosquitoes belonging to Anopheles atroparvus (n=171), Culex perexiguus (n=84), Culex pipiens (n=43), Culex theileri (n=39), Culex modestus (n=5), Ochlerotatus caspius (n=4), Culiseta sp. (n=1) were included in this study. Overall, hosts were identified from 234 blood meals compromising at least 25 species including mammals, birds and a single reptile. The success of host identification was lower in mosquitoes with an advanced stage of blood meal digestion and for blood meals extracted using the HotSHOT procedure.ConclusionsThe success of host identification decreases with the advanced stage of mosquito blood meal digestion, from 84.5% for recent blood meals to 25.0% for more digested ones. Using the QIAGEN kit, the identification success improved by 17.6%, with larger increases at more advanced stages of blood meal digestion. Availability of blood-fed females used to be very limited for studies of vector ecology, and these results may help to increase the efficiency of blood meal analyses. In addition, results obtained in this study clearly support that the potential malaria vector An. atroparvus feeds on animals located outdoors but use human-made shelters for resting after feeding.

Culicoids as Vectors of Schmallenberg Virus

Culicoids as Vectors of Schmallenberg Virus

Mosquito-specialist spiders

Mosquito-specialist spiders

Migration and motility of spermatozoa in the female reproductive tract of the soft tick Ornithodoros moubata (Acari, Argasidae)

The spermatozoa of ticks are anomalous in many respects: they are very large, cytoplasm-rich cells which lack a flagellum but move with a peculiar gliding motility. Their metamorphosis after deposition in the female has been well documented, but many of the subsequent events in the career of the spermatozoa are controversial or poorly documented. Our observations of motility imply that the many types of motility that have been reported (up to 5 different types in several reports) can be reduced to 2 apparently independent types of active motility: (1) gliding motility generated along the whole spermatozoon and (2) contortions of the anterior tip of the head. These types of motility appear as a consequence of sperm maturation after transfer to the female, but only become pronounced if the female has taken a recent bloodmeal. A consequence of this enhanced gliding motility after feeding is the movement of the spermatozoa out of the naturally ruptured neck of the spermatophore and up the female genital tract. This occurs without any apparent assistance from the female's musculature and likely is the prime mechanism of movement of the spermatozoa to the site of fertilization.

A spider that feeds indirectly on vertebrate blood by choosing female mosquitoes as prey.

Spiders do not feed directly on vertebrate blood, but a small East African jumping spider (Salticidae), Evarcha culicivora, feeds indirectly on vertebrate blood by choosing as preferred prey female mosquitoes that have had recent blood meals. Experiments show that this spider can identify its preferred prey by sight alone and by odor alone. When presented with two types of size-matched motionless lures, E. culicivora consistently chose blood-fed female mosquitoes in preference to nonmosquito prey, male mosquitoes, and sugar-fed female mosquitoes (i.e., females that had not been feeding on blood). When the choice was between mosquitoes of different sizes (both blood- or both sugar-fed), small juveniles chose the smaller prey, whereas adults and larger juveniles chose the larger prey. However, preference for blood took precedence over preference for size (i.e., to get a blood meal, small individuals took prey that were larger than the preferred size, and larger individuals took prey that were smaller than the preferred size). When presented with odor from two prey types, E. culicivora approached the odor from blood-fed female mosquitoes significantly more often the odor of the prey that were not carrying blood.

Feeding rates, nutritional status and flight dispersal potential of peridomestic populations of Triatomainfestans in rural northwestern Argentina

Triatoma infestans, the main vector of Chagas disease in Southern Cone countries, frequently infests peridomestic structures housing domestic animals. A total of 814 T. infestans collected from 35 different peridomestic sites in rural northwestern Argentina over 1–4 consecutive seasons was examined for recent blood meals and nutritional status. Bugs from goat or pig corrals had lower qualitative nutritional status and mean weight to length ratios ( W/L) than those captured in chicken coops. Males systematically had lower qualitative nutritional status and W/ L than females. Using logistic multiple regression, the daily feeding rates of T. infestans were significantly associated with season and stage but not ecotope, whereas the proportion of well-fed bugs varied significantly with all three factors. The seasonal trends in feeding rates and nutritional status were consistent with the local availability and breeding timing of domestic animals. The observed data fed into an empirical model predicted that the probability of flight initiation would peak in summer from pig or goat corrals, not chicken coops, and be insignificant in all ecotopes in spring and fall. Male T. infestans outnumbered females as potential fliers. This is the first study conducted in well-defined habitat units that shows significant heterogeneities in the feeding rates and nutritional status of triatomine populations linked to host demographics and management, and how these affect flight dispersal potential over seasons. Peridomestic bug populations are of great relevance as a source of domestic reinfestation and for the elimination of T. infestans.

Padronização da técnica imunoenzimática do ELISA de captura, no sistema avidina-biotina para a identificação de sangue ingerido por Lutzomyia (Lutzomyia) longipalpis (Lutz & Neiva, 1912)

Bloodmeals taken by insects constitute an important parameter for clarifying aspects of the transmission of zoonoses, including leishmaniases. Immunological assays can be used to investigate the attraction of vectors to animals, which may be hosts of these parasitoses. The objective of this study was to standardize a sandwich enzyme-linked immunosorbent assay and titer samples with different time periods of digestion, in laboratory-bred Lutzomyia longipalpis fed on rats. In the light of the high sensitivity that the biotin-avidin method permits, the technique provided at least ninety repeat tests for each sample and identified recent bloodmeals taken by these insects. Bloodmeals were detectable up to 12 and 24 h after blood ingestion, and a significant difference between these titers was observed.

Flagellate infections of Brazilian sand flies (Diptera: Psychodidae): isolation in vitro and biochemical identification of Endotrypanum and Leishmania.

Flagellate infections were found in 1,063 of 18,895 sand flies collected in the states of Amazonas, Pará, Rondonia and Acre, Brazil. Infection rates were 13.4% (species group Shannoni); 7.5% (subgenus Nyssomyia); 6.7% (subgenus Lutzomyia series Cruciata); 0.5% (genus Psychodopygus) and 3.1% for other sand flies (various subgenera). Leishmania braziliensis guyanensis and L. mexicana amazonensis were isolated, respectively, from the known vectors, Lutzomyia umbratilis and L. flaviscutellata. Single stocks of L. braziliensis-like and L. mexicana-like organisms were isolated, respectively, from L. whitmani and L. yuilli. Thirty-eight flagellate stocks, isolated by direct culture from sand flies were characterized in detail by morphology in culture, behavior in hamsters and mice and by enzyme profiles. Sixteen stocks from Lutzomyia sp. (Shannoni group) were identified as Endotrypanum schaudinni; 8 stocks from Lutzomyia sp. (Shannoni group) were identified as Endotrypanum sp.; 7 stocks from Psychodopygus ayrozai and P. paraensis were identified as Leishmania sp. previously isolated from the armadillo, Dasypus novemcinctus; 2 stocks of Trypanosoma rangeli were isolated from recently fed Lutzomyia sp. (Shannoni group) sand flies; the remaining 5 stocks from L. umbratilis and L. yuilli could not be identified. Observations suggested that Shannoni group sand flies were the natural vectors of Endotrypanum. Leishmania sp. infections in the man-biting flies P. ayrozai and P. paraensis were restricted to the midgut and associated with recent bloodmeals. Unidentified flagellates in L. umbratilis and L. yuilli were distributed throughout the digestive tract with no trace of bloodmeals.