Blood-fed Females Research Articles

Evaluation of a metofluthrin fan vaporizer device against phlebotomine sand flies (Diptera: Psychodidae) in a cutaneous leishmaniasis focus in the Judean Desert, Israel

The OFF! Clip-On fan vaporizer device releasing metofluthrin was evaluated against phletobomine sand flies in the Judean Desert, Israel, in October, 2009. A total of 76,400 sand flies was collected, with male flies representing 98.3%Phlebotomus sergenti and 1.7%P. papatasi. Females comprised 43.0% of the total catch and included 6.7% blood-fed females. Similar proportions of flies were collected in both suction and sticky traps. In trials with unbaited suction traps, similar numbers of sand flies were collected in traps with a metofluthrin device, blank device, or no device (i.e., suction only). In suction traps baited with CO(2) , higher numbers of P. sergenti males and blood-fed females were collected in traps with a blank device compared to traps with a metofluthrin device. In sticky traps baited with CO(2) , there were no significant differences between catches in traps with a metofluthrin device, blank device, or no device. The results suggest metofluthrin from the device is not repellent against sand flies in a field environment despite showing insecticidal activity against flies collected in suction traps.

Site-Specific Integration and Expression of an Anti-Malarial Gene in Transgenic Anopheles gambiae Significantly Reduces Plasmodium Infections

Diseases transmitted by mosquitoes have a devastating impact on global health and this is worsening due to difficulties with existing control measures and climate change. Genetically modified mosquitoes that are refractory to disease transmission are seen as having great potential in the delivery of novel control strategies. Historically the genetic modification of insects has relied upon transposable elements which have many limitations despite their successful use. To circumvent these limitations the Streptomyces phage phiC31 integrase system has been successfully adapted for site-specific transgene integration in insects. Here, we present the first site-specific transformation of Anopheles gambiae, the principal vector of human malaria. Mosquitoes were initially engineered to incorporate the phiC31 targeting site at a defined genomic location. A second phase of genetic modification then achieved site-specific integration of Vida3, a synthetic anti-malarial gene. Expression of Vida3, specifically in the midgut of bloodfed females, offered consistent and significant protection against Plasmodium yoelii nigeriensis, reducing average parasite intensity by 85%. Similar protection was observed against Plasmodium falciparum in some experiments, although protection was inconsistent. In the fight against malaria, it is imperative to establish a broad repertoire of both anti-malarial effector genes and tissue-specific promoters for their expression, enabling those offering maximum effect with minimum fitness cost to be identified. In the future, this technology will allow effective comparisons and informed choices to be made, potentially leading to complete transmission blockade.

Recent field studies on vector ecology of mosquitoes in urban areas of Tokyo, Japan

Results of two mosquito studies recently conducted in a park in urban Tokyo were introduced and related studies were reviewed. A large number of adult Culex tritaeniorhynchus suddenly appeared from late September to November 2007 during the course of systematic sweep net collections of vegetation in the park. The pre-diapause migration has been observed every year from 2007 to 2010. The time course of pre-diapause migration was similar starting in mid-/late September, peaking in mid-October and ending in December. The density of migrated females and parity differed year by year, and dissection showed that a large part of the females (85 to 97%) were in reproductive diapause. Blood-meal identifications of Cx. pipiens pallens together with avian malaria parasite detection were conducted by using blood-fed females collected in the study park from May 2007 to May 2008. Thirteen wild bird species were identified as the blood-source and 5 of them were infected with avian malaria parasite. Ten Plasmodium lineages were identified and the three commonest lineages were detected in both the abdomens and thoraxes of mosquitoes, suggested transmission of these lineages. Jungle crow and tree sparrow served as a natural host for the three commonest lineages.

Comparison of the Physiological age of Adult Resting Versus Flying Culex Quinquefasciatus (Diptera: Culicidae)

Relatively little is known about the physiological age of resting mosquitoes. In this study, Culex quinquefasciatus Say had a similar proportion of males (75%) in the resting as in the flying population (76%), but among the females, there were significant differences between the physiological age of the resting and flying populations. The proportion of unfed females (51%) was significantly smaller in the resting than in the flying female group (82%). By contrast, in the resting group, there were much higher proportions of blood-fed (28% compared with 4%) and to a lesser extent gravid (21% compared with 14%) females. Of the blood-fed females, most of the resting individuals were freshly blood-fed. Both males and females entered the resting traps throughout the day (but none were collected after sunset), with the main peak around dawn. The flying population showed two circadian peaks: a larger peak at dawn and smaller peak after sunset. This indicates a different pattern from that shown in other studies and demonstrates the significant geographic variation within this pan-tropical species.

Effect of Sublethal Dose of Diflubenzuron and Azadirachtin on Various Life Table Attributes of Culex quinquefasciatus (Diptera: Culicidae)

The exposure of sublethal doses (25% emergence inhibition) of diflubenzuron (DFB) (0.0002 mg/liter) and azadirachtin (AZD; 0.0079 mg/liter) insect growth regulators was provided to late third- early fourth instars of Culex quinquefasciatus till emergence of adults to find out the effects on various biological and life table attributes of adults, and comparison was made with control (CTL) as well. Life expectancy of both males and females of DFB survivors was significantly less in comparison with AZD and CTL ones; however, it was nonsignificantly different between CTL and AZD. In all treatments, males were shorter lived than females of their respective treatments. DFB survivors showed significant reduction in number of blood-fed females, resulting in minimum of oviposition days, total number of egg rafts, total number of eggs, eggs per egg raft, eggs per female life span, and eggs per female per day than survivors of both AZD and CTL significantly, but these parameters did not differ significantly between CTL and AZD, except eggs per egg raft. The net reproductive rate, intrinsic rate of increase, and finite rate of increase were significantly less in DFB than rest of the survivors, whereas doubling time for generation was significantly high; however, difference in mean generation was not significant among them. This study reveals that DFB has the potency to affect severely the various life table attributes at sublethal dose, but AZD does not have such effectiveness at similar sublethal dose against Cx. quinquefasciatus.

Contribution to the knowledge of Culicoides (Diptera: Ceratopogonidae) host preferences in France

Knowledge on host-feeding pattern of blood-sucking insects helps to understand the epidemiology of a vector-born disease. We determined blood meal origin from blood-fed Culicoides thanks to molecular techniques. A set of primers was used to selectively amplify segment of vertebrates' prepronociceptin gene from abdomen of engorged Culicoides (Diptera: Ceratopogonidae). Vertebrate DNA was successfully amplified in 91% of blood-fed Culicoides assayed. Direct sequencing and comparison of resultant sequences with sequences in GenBank, using BLAST, lead to the specific identification of the host in 100% of the cases. A total of 157 blood-fed females belonging to 13 different Culicoides' species were captured thanks to light traps in different areas of France between 2008 and 2009. Blood meal origin was determined for 143 blood-fed midges: 59 Culicoides obsoletus, 18 Culicoides dewulfi, 16 Culicoides scoticus, 11 Culicoides chiopterus, 10 Culicoides lupicaris, 1 Culicoides pulicaris, 8 Culicoides punctatus, 10 Culicoides pallidicornis, 3 Culicoides achrayi, 2 Culicoides furcillatus, 3 Culicoides brunnicans, 1 Culicoides picturatus and 1 Culicoides poperinghensis. The predominant species in our study belong to the C. obsoletus complex; they are considered as putative vectors of Bluetongue virus in the north of Europe. C. chiopterus sampled fed only on cattle, while blood meal origin of C. dewulfi, C. obsoletus and C. scoticus was diversified. In our sampling, we found that midges were fed mainly on cattle (54%), rabbits (20%), horses (17%), sheep (4%), pigs or wild boars (4%) and human (1%). Cattle DNA was found in at least 11 different species of Culicoides assayed.

Two insulin-like peptide family members from the mosquito Aedes aegypti exhibit differential biological and receptor binding activities

Insects encode multiple ILPs but only one homolog of the vertebrate IR that activates the insulin-signaling pathway. However, it remains unclear whether all insect ILPs are high affinity ligands for the IR or have similar biological functions. The yellow fever mosquito, Aedes aegypti, encodes eight ILPs with prior studies strongly implicating ILPs from the brain in regulating metabolism and the maturation of eggs following blood feeding. Here we addressed whether two ILP family members expressed in the brain, ILP4 and ILP3, have overlapping functional and receptor binding activities. Our results indicated that ILP3 exhibits strong insulin-like activity by elevating carbohydrate and lipid storage in sugar-fed adult females, whereas ILP4 does not. In contrast, both ILPs exhibited dose-dependent gonadotropic activity in blood-fed females as measured by the stimulation of ovaries to produce ecdysteroids and the uptake of yolk by primary oocytes. Binding studies using ovary membranes indicated that ILP4 and ILP3 do not cross compete; a finding further corroborated by cross-linking and immunoblotting experiments showing that ILP3 binds the MIR while ILP4 binds an unknown 55 kDa membrane protein. In contrast, each ILP activated the insulin-signaling pathway in ovaries as measured by enhanced phosphorylation of Akt. RNAi and inhibitor studies further indicated that the gonadotropic activity of ILP4 and ILP3 requires the MIR and a functional insulin-signaling pathway. Taken together, our results indicate that two members of the Ae. aegypti ILP family exhibit partially overlapping biological activity and different binding interactions with the MIR.

Sampling Biases of the BG-Sentinel Trap With Respect to Physiology, Age, and Body Size of Adult Aedes aegypti (Diptera: Culicidae)

AbstractCurrently, Aedes aegypti (L.) control strategies are being developed that involve manipulation of the vector at the adult stage (e.g., the use of the bacterial endosymbiont Wolbachia to shorten the life span of the vector population). These novel strategies demand adult sampling methods to measure changes in population size, structure (age, sex ratio), and, ultimately, the success of the program. Each sampling method presents certain biases. Once these biases are defined, methods used to estimate population size and structure can be calibrated accordingly, resulting in more accurate and complex estimates of the vector population. A series of mark-release-recapture experiments with adult Ae. aegypti were conducted in a large outdoor flight cage and an indoor setting in far north Queensland, Australia. The biases of the BG-Sentinel trap (BGS) were investigated across several categories, as follows: 1) mosquito age; 2) sex; 3) physiological status; and 4) body size. Biases were not detected across age groups or body sizes. A significant bias was detected across physiological groups: nulliparous females were recaptured at a significantly lower rate than all other groups except blood-fed parous females, which were also recaptured at a low rate by the BGS. Males were recaptured at a higher rate than all groups, but only a significant difference in recapture rates was observed between males and nulliparous females. Previous studies show that the BGS is a highly effective tool for Ae. aegypti surveillance. The BGS proves to be a reliable tool in Ae. aegypti surveillance with consistent sampling outcomes. The sampling bias of the BGS is measurable and can be used to generate more accurate estimates of the adult population and its attributes.

Sampling Biases of the BG-Sentinel Trap With Respect to Physiology, Age, and Body Size of Adult <I>Aedes aegypti</I> (Diptera: Culicidae)

Currently, Aedes aegypti (L.) control strategies are being developed that involve manipulation of the vector at the adult stage (e.g., the use of the bacterial endosymbiont Wolbachia to shorten the life span of the vector population). These novel strategies demand adult sampling methods to measure changes in population size, structure (age, sex ratio), and, ultimately, the success of the program. Each sampling method presents certain biases. Once these biases are defined, methods used to estimate population size and structure can be calibrated accordingly, resulting in more accurate and complex estimates of the vector population. A series of mark-release-recapture experiments with adult Ae. aegypti were conducted in a large outdoor flight cage and an indoor setting in far north Queensland, Australia. The biases of the BG-Sentinel trap (BGS) were investigated across several categories, as follows: 1) mosquito age; 2) sex; 3) physiological status; and 4) body size. Biases were not detected across age groups or body sizes. A significant bias was detected across physiological groups: nulliparous females were recaptured at a significantly lower rate than all other groups except blood-fed parous females, which were also recaptured at a low rate by the BGS. Males were recaptured at a higher rate than all groups, but only a significant difference in recapture rates was observed between males and nulliparous females. Previous studies show that the BGS is a highly effective tool for Ae. aegypti surveillance. The BGS proves to be a reliable tool in Ae. aegypti surveillance with consistent sampling outcomes. The sampling bias of the BGS is measurable and can be used to generate more accurate estimates of the adult population and its attributes.

Effects of Life Stages and Feeding Regimes on Active Movement Behavior of the Tropical Bed Bug, <I>Cimex hemipterus</I> (Hemiptera: Cimicidae)

This study examined the effects of different life stages (first, second, third, fourth, and five instars; adult females and adult males) and feeding regimes (starved and blood fed) on the active movement activity of the tropical bed bug, Cimex hemipterus (F.), under mixed-stage conditions. We used an extended arena made from Tygon tube coils and observed the movement frequency and movement distance at selected time intervals up to 120 h. The fifth instars and adult males and females showed significantly (P < 0.01) greater movement frequency compared with the other stages. The first and second instars showed limited movement (< 8 m) over the experimental period. Starved bed bugs showed greater movement frequency compared with blood-fed bed bugs, with the exception of adult females. Blood-fed adult females exhibited significantly (P < 0.01) greater movement frequency and distance compared with starved females. Blood-fed females moved up to 42.3 m over 120 h. Regression analysis between movement distance of the fifth instars and adults and the time intervals revealed a positive relationship (r2 = 0.6583; P < 0.01), suggesting that delays in bed bug control efforts will increase the risk of the greater infestation. During bed bug inspection, the presence of only late instars and adults in premises would indicate a new infestation, whereas an established infestation likely would consist of mixed stages.

The RNA interference pathway affects midgut infection- and escape barriers for Sindbis virus in Aedes aegypti

BackgroundThe RNA interference (RNAi) pathway acts as an innate antiviral immune response in Aedes aegypti, modulating arbovirus infection of mosquitoes. Sindbis virus (SINV; family: Togaviridae, genus: Alphavirus) is an arbovirus that infects Ae. aegypti in the laboratory. SINV strain TR339 encounters a midgut escape barrier (MEB) during infection of Ae. aegypti. The nature of this barrier is not well understood. To investigate the role of the midgut as the central organ determining vector competence for arboviruses, we generated transgenic mosquitoes in which the RNAi pathway was impaired in midgut tissue of bloodfed females. We used these mosquitoes to reveal effects of RNAi impairment in the midgut on SINV replication, midgut infection and dissemination efficiencies, and mosquito longevity.ResultsAs a novel tool for studying arbovirus-mosquito interactions, we engineered a transgenic mosquito line with an impaired RNAi pathway in the midgut of bloodfed females by silencing expression of the Aa-dcr2 gene. In midgut tissue of the transgenic Carb/dcr16 line, Aa-dcr2 expression was reduced ~50% between 1-7 days post-bloodmeal (pbm) when compared to the recipient mosquito strain. After infection with SINV-TR339EGFP, Aa-dcr2 expression levels were enhanced in both mosquito strains. In the RNAi pathway impaired mosquito strain SINV titers and midgut infection rates were significantly higher at 7 days pbm. There was also a strong tendency for increased virus dissemination rates among the transgenic mosquitoes. Between 7-14 days pbm, SINV was diminished in midgut tissue of the transgenic mosquitoes. Transgenic impairment of the RNAi pathway and/or SINV infection did not affect longevity of the mosquitoes.ConclusionsWe showed that RNAi impaired transgenic mosquitoes are a useful tool for studying arbovirus-mosquito interactions at the molecular level. Following ingestion by Ae. aegypti, the recombinant SINV-TR339EGFP was confronted with both MEB and a midgut infection barrier (MIB). Impairment of the RNAi pathway in the midgut strongly reduced both midgut barriers for the virus. This confirms that the endogenous RNAi pathway of Ae. aegypti modulates vector competence for SINV in the midgut. The RNAi pathway acts as a gatekeeper to the incoming virus by affecting infection rate of the midgut, intensity of infection, and dissemination from the midgut to secondary tissues.

Host-Feeding Patterns ofAedes albopictus(Diptera: Culicidae) in Urban and Rural Contexts within Rome Province, Italy

Knowledge of the frequency of contact between a mosquito species and its different hosts is essential to understand the role of each vector species in the transmission of diseases to humans and/or animals. However, no data are so far available on the feeding habits of Aedes albopictus in Italy or in other recently colonized temperate regions of Europe, due to difficulties in collecting blood-fed females of this diurnal and exophilic species. We analyzed Ae. albopictus host-feeding patterns in two urban and two rural sites within the area of Rome (Italy). Ae. albopictus was collected using sticky-traps and the blood-meal origin of 303 females was determined by direct dot-ELISA. The blood-fed sample, although representing only 4% of the total Ae. albopictus collected, demonstrates the useful application of sticky-trap in studying the feeding behavior of the species. The human blood index was significantly different among sites, ranging from 79-96% in urban sites to 23-55% in rural sites, where horses and bovines represented the most bitten hosts. The results obtained confirm the plastic feeding behavior shown by Ae. albopictus in its original range of distribution and highlights the high potential of this species as a vector of human pathogens in urban areas of Italy, where both humans and the mosquito itself may reach very high densities.

Direct sequencing and expression analysis of a large number of miRNAs in Aedes aegypti and a multi-species survey of novel mosquito miRNAs

BackgroundMicroRNAs (miRNAs) are a novel class of gene regulators whose biogenesis involves hairpin structures called precursor miRNAs, or pre-miRNAs. A pre-miRNA is processed to make a miRNA:miRNA* duplex, which is then separated to generate a mature miRNA and a miRNA*. The mature miRNAs play key regulatory roles during embryonic development as well as other cellular processes. They are also implicated in control of viral infection as well as innate immunity. Direct experimental evidence for mosquito miRNAs has been recently reported in anopheline mosquitoes based on small-scale cloning efforts.ResultsWe obtained approximately 130, 000 small RNA sequences from the yellow fever mosquito, Aedes aegypti, by 454 sequencing of samples that were isolated from mixed-age embryos and midguts from sugar-fed and blood-fed females, respectively. We also performed bioinformatics analysis on the Ae. aegypti genome assembly to identify evidence for additional miRNAs. The combination of these approaches uncovered 98 different pre-miRNAs in Ae. aegypti which could produce 86 distinct miRNAs. Thirteen miRNAs, including eight novel miRNAs identified in this study, are currently only found in mosquitoes. We also identified five potential revisions to previously annotated miRNAs at the miRNA termini, two cases of highly abundant miRNA* sequences, 14 miRNA clusters, and 17 cases where more than one pre-miRNA hairpin produces the same or highly similar mature miRNAs. A number of miRNAs showed higher levels in midgut from blood-fed female than that from sugar-fed female, which was confirmed by northern blots on two of these miRNAs. Northern blots also revealed several miRNAs that showed stage-specific expression. Detailed expression analysis of eight of the 13 mosquito-specific miRNAs in four divergent mosquito genera identified cases of clearly conserved expression patterns and obvious differences. Four of the 13 miRNAs are specific to certain lineage(s) within mosquitoes.ConclusionThis study provides the first systematic analysis of miRNAs in Ae. aegypti and offers a substantially expanded list of miRNAs for all mosquitoes. New insights were gained on the evolution of conserved and lineage-specific miRNAs in mosquitoes. The expression profiles of a few miRNAs suggest stage-specific functions and functions related to embryonic development or blood feeding. A better understanding of the functions of these miRNAs will offer new insights in mosquito biology and may lead to novel approaches to combat mosquito-borne infectious diseases.

Aspects of the bionomics of Phlebotomus sergenti sandflies from an endemic area of anthroponotic cutaneous leishmaniasis in Aleppo Governorate, Syria

Aspects of the bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied from June to November 2005 in three foci of anthroponotic cutaneous leishmaniasis (ACL) in Aleppo Governorate, Syria, where the agent Leishmania tropica (Wright) (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus sergenti Parrot. Syria has been designated by the World Health Organization as one of four countries in the Old World where cutaneous leishmaniasis is hyperendemic, but little is known about the biology of local vector populations. Standard collections by sticky traps showed two peaks in density, in June and late August. In total, 1840 sandflies were caught, comprising five species: Phlebotomus papatasi (Scopoli) (68.0%); P. sergenti (25.4%); Sergentomyia minuta (Rondani) (6.4%); Phlebotomus tobbi Adler & Theodor (0.1%), and Phlebotomus mascittii canaaniticus Adler & Theodor (0.1%). Similar numbers of P. sergenti were caught indoors (246 specimens) and outdoors (222), whereas P. papatasi was significantly more abundant indoors (1096 specimens) than outdoors (156) (chi(2) = 241, P < 0.01). In total, 212 blood-fed females were tested for host blood determination, of which 176 (83.0%) reacted with anti-species reagent. Results from 20 P. sergenti suggest that this species is an opportunistic feeder, imbibing human, ovine, avian, bovine and feline blood, although more bloodmeals were taken from humans and cattle than expected in relation to the relative proportions of potential hosts present (the forage ratio, FR). The bionomics of P. sergenti are discussed in relation to the inefficacy of control campaigns based on indoor spraying with residual insecticides that have been implemented by the Syrian Ministry of Health to control the epidemics of ACL in the Aleppo Governorate.

Insecticide resistance profiles for malaria vectors in the Kassena-Nankana district of Ghana

BackgroundMalaria is a major public health problem in Ghana. The current strategy of the National Malaria Control Programme is based on effective case management and the use of insecticide treated bed nets among vulnerable groups such as children under-five years of age and pregnant women. Resistance to pyrethroids by Anopheles gambiae s.l. and Anopheles funestus has been reported in several African countries including neighbouring Burkina Faso.MethodsIndoor resting Anopheles mosquitoes were collected. Blood-fed and gravid females were allowed to oviposit, eggs hatched and larvae reared to 1–3 days old adults and tested against permethrin 0.75%, deltamethrin 0.05%, cyfluthrin 0.15%, lambdacyhalothrin 0.1% and DDT 4%, based on WHO methodology. PCR analyses were carried out on a sub-sample of 192 of the An. gambiae for sibling species complex determination. Resistance to pyrethroids and DDT was determined by genotyping the knock-down resistance kdr gene mutations in the study area.ResultsA total of 9,749 1–3 days-old F1 female Anopheles mosquitoes were exposed to the insecticides. Among the pyrethroids, permethrin, 0.75% had the least knockdown effect, whilst cyfluthrin 0.15%, had the highest knock-down effect. Overall, no difference in susceptibility between An. gambiae 93.3% (95% CI: 92.5–94.1) and An. funestus 94.5% (95% CI: 93.7–95.3) was observed when exposed to the pyrethroids. Similarly, there was no difference in susceptibility between the two vector species (An. gambiae = 79.1% (95% CI: 76.6–81.8) and An. funestus = 83.5% (95% CI: 80.2–86.4) when exposed to DDT. Overall susceptibility to the insecticides was between 80% and 98%, suggesting that there is some level of resistance, except for cyfluthrin 0.15%. The kdr PCR assay however, did not reveal any kdr mutations. The analysis also revealed only the molecular M (Mopti) form.ConclusionThe findings in this study show that An. gambiae and An. funestus, the main malaria vector mosquitoes in the Kassena-Nankana district are susceptible to the insecticides being used in the treatment of bed nets in the malaria control programme. There is however, the need for continuous monitoring of the pyrethroids as the efficacy is not very high.

Novel use of stir bar sorptive extraction (SBSE) as a tool for isolation of oviposition site attractants for gravid Culex quinquefasciatus

Mosquitoes such as Culex quinquefasciatus Say (Diptera: Culicidae) are important vectors of organisms that cause disease in humans. Research into the development of effective standardized odour baits for blood-fed females (oviposition attractants), to enable entomological monitoring of vector populations, is hampered by complex protocols for extraction of physiologically active volatile chemicals from natural breeding site water samples, which have produced inconsistent results. Air entrainment and solvent extraction are technically demanding methods and are impractical for use in resource poor environments where mosquito-borne disease is most prevalent. This study reports the first use of a simple, robust extraction technique, stir bar sorptive extraction (SBSE), to extract behaviourally active small lipophilic molecules (SLMs) present in water samples collected from Cx. quinquefasciatus breeding sites in Tanzania. Extracts from a pit latrine and from a cess pool breeding site attracted more gravid Cx. quinquefasciatus in pair choice bioassays than control extracts, and coupled gas chromatography-electroantennography (GC-EAG) allowed tentative identification of 15 electrophysiologically active chemicals, including the known oviposition attractant, skatole (3-methylindole). Here, we have demonstrated, using simple pair choice bioassays in controlled laboratory conditions, that SBSE is effective for the extraction of behaviourally and electrophysiologically active semiochemicals from mosquito breeding site waters. Further research is required to confirm that SBSE is an appropriate technique for use in field surveys in the search for oviposition cues for Cx. quinquefasciatus.

Surveillance des moucherons piqueurs : détermination des périodes sans vecteurs en Suisse de 2007 à 2009

Since the introduction of bluetongue virus serotype 8 (BTV-8) in Northern Europe, a surveillance of the disease has been imple­mented in Switzerland, based on a collaborative network includ­ing the Swiss Federal Veterinary Office, the Institute of Virology and Immunoprophylaxis and the Institute of Parasitology (IPZ). In accordance with European regulations, the surveillance aims at determining zones which are seasonally free of bluetongue by showing lack of evidence of disease transmission or of activity of competent vectors. The results of two years (2007–2009) of sur­veillance of the winter activity of biting midges are presented. BT zones were identified for Switzerland and Liechtenstein based on (i) the 45x45 km grid, (ii) a spatial risk analysis on the occur­rence of biting midges and susceptible livestock, and (iii) the limits of the Swiss administrative units (cantons). Thus, 16 BT zones were defined, comparable in area size (as ‘risk’ areas) and in populations of cattle, sheep and goats. Insects were caught with Onderstepoort ultraviolet-light traps once weekly at 19 sta­tions, outdoors, from dusk to dawn, and from October to May. Trapping was maintained at six stations during the midge activ­ity season. Trappings were implemented by volunteer farmers or local veterinary offices, and samples were sent to the Swiss reference laboratory for vector entomology (IPZ) where midge activity was classified from ‘no activity’ to ‘very high activity’. Midges were caught at all stations, but in very different numbers. From mid-November 2007 until end of March 2008 there was a period of very low Culicoides activity (catches without midges or below the threshold of 10 midges/trap/night). Then, the activity increased gradually, and the vector-free period was defined as ending on April 15th. All sites showed activity during the whole summer, globally with a decrease from July to September. The highest activity was recorded in Northern Switzerland (Basel), with an average of 3268 midges/trap/night. Surprisingly, the highest activity was not observed in Southern Switzerland where the mild climate is influenced by the Mediterranean Sea, but north of the Alps characterized by an Atlantic climate. During October to mid-November 2008, midge activity decreased, and less than 10 midges or 5 parous midges/trap/night were caught at all sites after mid-November. The vector-free period was thus determined as starting on December 15, 2008. Then, a very low activity was observed in Basel during early January, and no activity was observed in the cantons until the first week of March 2009. Very few midges were trapped until beginning of April, and first parous and blood-fed females were caught after mid- April. Thus, the vector-free period was declared to end on April 20, 2009. Vector-free periods could be defined for the whole of Switzerland during the two years of winter surveillance. A third year of surveillance is ongoing, and three-years’ data will be ana­lysed for a possible extrapolation of an annual vector-free period.

Use of Phlebotomine Sand Flies as Indicator of Leishmania Prevalence in an Endemic Area

An entomological survey was conducted on phlebotomine sand flies to discover the presence of Leishmania parasites in six Sicilian provinces. During the season from June to October 2005, 20,346 sand fly specimens were collected by using a "black light" trap in eight cattle and sheep farms. The most prevalent species was Phlebotomus perfiliewi (51.60%) followed by P. perniciosus (24.37%), and Sergentomyia minuta (24.03%). Among the 11,441 females collected, 284 (2.70%) were gravid and 708 (6.74%) were blood-fed females. A total of 194 gravid females, 274 blood-fed females, and 254 females with no evidence of blood meal were analyzed for L. infantum by PCR analysis, showing 9.79% gravid females, 5.84% blood-fed females, and 5.51% females with no evidence of blood meal positive for Leishmania DNA. These results support the use of gravid female sand flies for the evaluation of prevalence of Leishmania.

Structural Changes in Fat Body of Aedes aegypti Caused by Aging and Blood Feeding

The fat body is the intermediary metabolism organ of insects and the main source of hemolymph components. In the current study, the microanatomy of Aedes aegypti (L., 1762) fat body was studied through scanning electron microscopy to observe the effects of blood feeding and aging. Three groups of female mosquitoes were used: newly emerged females, 18-d-old sugar-fed females, and 18-d-old blood-fed females. In Ae. aegypti, the fat body is located beneath the integument, and it is subdivided into dorsal, ventral, and lateral lobes, with the latter two being larger than the dorsal lobes. The lobes projected into the body cavity, and they were covered externally by a basal lamina with rounded cells beneath it. In 18-d-old sugar-fed females, the ventral and dorsal fat bodies seemed more developed than in newly emerged mosquitoes. The fat body hypertrophy caused by aging in the sugar-fed mosquito was probably associated with lipid accumulation due to the sugar diet. The blood-fed 18-d-old mosquitoes showed flattened fat bodies in all locations. The fat body modifications after the blood ingestion may be associated with midgut expansion after blood feeding, followed by ovary hypertrophy that mechanically compresses the fat body against the body wall. The structural changes in the fat body after a bloodmeal may be important for midgut extension to maximize blood storage and subsequent ovary enlargement, leading to the organ’s reorganization in the body cavity. In addition, the depletion of fat body content during vitellogenesis could be responsible for the shrinking and flattening of the fat body lobes.

Structural Changes in Fat Body of &lt;I&gt;Aedes aegypti&lt;/I&gt; Caused by Aging and Blood Feeding

The fat body is the intermediary metabolism organ of insects and the main source of hemolymph components. In the current study, the microanatomy of Aedes aegypti (L., 1762) fat body was studied through scanning electron microscopy to observe the effects of blood feeding and aging. Three groups of female mosquitoes were used: newly emerged females, 18-d-old sugar-fed females, and 18-d-old blood-fed females. In Ae. aegypti, the fat body is located beneath the integument, and it is subdivided into dorsal, ventral, and lateral lobes, with the latter two being larger than the dorsal lobes. The lobes projected into the body cavity, and they were covered externally by a basal lamina with rounded cells beneath it. In 18-d-old sugar-fed females, the ventral and dorsal fat bodies seemed more developed than in newly emerged mosquitoes. The fat body hypertrophy caused by aging in the sugar-fed mosquito was probably associated with lipid accumulation due to the sugar diet. The blood-fed 18-d-old mosquitoes showed flattened fat bodies in all locations. The fat body modifications after the blood ingestion may be associated with midgut expansion after blood feeding, followed by ovary hypertrophy that mechanically compresses the fat body against the body wall. The structural changes in the fat body after a bloodmeal may be important for midgut extension to maximize blood storage and subsequent ovary enlargement, leading to the organ's reorganization in the body cavity. In addition, the depletion of fat body content during vitellogenesis could be responsible for the shrinking and flattening of the fat body lobes.