The mosquitoes of Sardinia: species records 35 years after the malaria eradication campaign

1. Thirty-five years ago malaria was eradicated from Sardinia by massive application of DDT directed against the mosquito vector species, Anopheles labranchiae, but the long-term effects on the mosquito fauna have not been investigated previously. 2. A 5-year mosquito survey (1980-85) involved in the inspection of 348 sites (plus 173 reinspections) and the processing of nearly 60,000 specimens. 3. Twenty-four species in six genera were identified from larval and adult collections. Species of the genera Culex (41%) and Aedes (22%) were more commonly encountered than Culiseta (18%) and Anopheles (17%) species. Orthopodomyia (1.6%) and Coquillettidia (0.6%) were far less frequent. 4. The species composition was similar to that described for the years of the eradication campaign, except for the genus Anopheles. Of the eight Anopheles species previously recorded, only four species were identified in collections of 1159 larvae from eighty-five positive sites. 5. Anopheles labranchiae was the most frequently collected species of its genus (49%), but was represented in only 12% of the total collections. Comparison with previous records indicated a progressive increase in sites positive for An.labranchiae over the past 35 years. 6. Anopheles hispaniola was not found, although it had apparently replaced An.labranchiae soon after the eradication campaign. An. labranchiae was common in the sites where the replacement phenomenon has been observed.     

Influence of resource levels,organic compounds and laboratory colonization on interspecific competition between the Asian tiger mosquito Aedes albopictus (Stegomyia albopicta) and the southern house mosquito Culex quinquefasciatus

The mosquitoes Aedes albopictus (Stegomyia albopicta) (Skuse) and Culex quinquefasciatus (Say) (Diptera: Culicidae) are common inhabitants of tyres and other artificial containers, which constitute important peridomestic mosquito breeding habitats. We tested the hypotheses that interspecific resource competition between the larvae of these species is asymmetrical, that the concentration of chemicals associated with decomposing detritus affects the competitive outcomes of these species, and that wild and colonized strains of Cx. quinquefasciatus are affected differently by competition with Ae. albopictus. We conducted two laboratory competition experiments wherein we measured survivorship and estimated population growth (λ′) in both species under multiple mixed‐species densities. Under varying resource levels, competition was asymmetrical: Ae. albopictus caused competitive reductions or exclusions of Cx. quinquefasciatus under conditions of limited resources. In a second experiment, which used both wild and colonized strains of Cx. quinquefasciatus, organic chemical compounds associated with decomposing detritus did not affect the competitive outcome. The colonized strain of Cx. quinquefasciatus had greater survivorship and adult mass, and faster development times than the wild strain, but both strains were similarly affected by competition with Ae. albopictus. Competition between these species may have important consequences for vector population dynamics, especially in areas in which tyres and artificial containers constitute the majority of mosquito breeding habitats.     

West Nile virus cluster analysis and vertical transmission in Culex pipiens complex mosquitoes in Sacramento and Yolo Counties,California, 2011

West Nile virus (WNV) is now endemic in California, with annual transmission documented by the statewide surveillance system. Although much is known about the horizontal avian‐mosquito transmission cycle, less is known about vertical transmission under field conditions, which may supplement virus amplification during summer and provide a mechanism to infect overwintering female mosquitoes during fall. The current study identified clusters of WNV‐infected mosquitoes in Sacramento and Yolo Counties, CA, during late summer 2011 and tested field‐captured ovipositing female mosquitoes and their progeny for WNV RNA to estimate the frequency of vertical transmission. Space‐time clustering of WNV‐positive Culex pipiens complex pools was detected in the northern Elk Grove area of Sacramento County between July 18 and September 18, 2011 (5.22 km radius; p<0.001 and RR=7.80). Vertical transmission by WNV‐infected females to egg rafts was 50% and to larvae was 40%. The estimated minimal filial infection rate from WNV‐positive, ovipositing females was 2.0 infected females/1,000. The potential contribution of vertical transmission to WNV maintenance and amplification are discussed.     

Experimental infection of Aedes (Stegomyia) albopictus and Culex pipiens mosquitoes with Bluetongue virus

Bluetongue disease (BT), caused by Bluetongue virus (BTV), infects wild and domestic ruminants, causing severe economic damage in the cattle and sheep industry. Proven vectors of BTV are biting midges belonging to the Culicoides genus, but other arthropods are considered potential vectors, such as ticks, mosquitoes, wingless flies, and sand flies. The present study represents the first attempt to evaluate the vectorial capacity of Culex pipiens and Aedes albopictus for BTV. Mosquitoes were artificially fed with blood containing BTV serotype 1. Infection, dissemination and transmission rates were evaluated at 0, 3, 7, 14 and 21 days after an infected blood meal. Viral RNA was only detected up to 3 days post infection in the bodies of both species. This study indicates that the two Italian populations of Cx. pipiens and Ae. albopictus are not susceptible to BTV infection.     

中国库蚊属库状蚊亚属一新种：双翅目：蚊科

本文记述库蚊属库状蚊亚属一新种——刺端库蚊,新种Culez(Culiciomyia) spiculostylussp.nov.。本新种阳茎侧板背角短粗而端钝,密布小齿并形成明显的齿簇为其独具特征。模式标本系1985年8月11日采自云南省西双版纳勐腊县新公社一积水坑,存放于贵阳医学院生物学教研室。     

Resistance potential of Culex quinquefasciatus against the insect growth regulators methoprene and diflubenzuron

Bioassay procedures for the insect growth regulators diflubenzuron (a chitin synthetase inhibitor) and methoprene (a juvenile hormone mimic) were evaluated on the mosquito Culex quinquefasciatus Say to determine the most sensitive larval stages and the effects of varying exposure periods. Sensitivity to diflubenzuron decreased gradually during instars I and II; instars III and IV were relatively insusceptible. Sensitivity to methoprene increased with larval age. Standard test procedures were devised for diflubenzuron involving continuous exposure from the early third instar and for methoprene as 6 h exposure during instar IV. Selection for ten generations of strains from Dar-es-Salaam which were multiresistant to insecticides increased the LC₅₀ to diflubenzuron by 2.8-fold and increased the LC₅₀ to methoprene by 3.9-fold. When compared to a standard susceptible strain from Colombo the resistance ratios of Dar-es-Salaam strains rose from 2.4 x to 6.6 x for diflubenzuron and from 6.1 x to 21.3 x for methoprene, with no significant effects detected on the pattern of cross-resistance. These results are discussed in relation to standardization of IGR testing and the need for integrated control programmes against Cx quinquefasciatus.

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Résistance potentielle de Culex quinquefasciatus Say au méthoprène et au diflubenzuron, régulateurs de croissance des insectes

Résumé La détermination du stade larvaire le plus sensible et les effects de différentes durées d'exposition aux régulateurs de croissance des insectes-diflubenzuron (inhibiteur de la chitine synthétase) et méthoprène (mimétique de l'hormone juvénile)—ont été recherchés sur C. quinquefasciatus. La sensibilité au diflubenzuron diminue graduellement pendant les deux premiers stades, les stades III et IV étant relativement insensibles. La sensibilité au méthoprène augmente avec l'âge des larves. Les essais comportaient une exposition continue au diflubenzuron depuis le début du stade III et une exposition de 6 heures au méthoprène pendant le stade IV. La sélection pendant 10 générations de lignées de Dares-Salam multirésistantes aux insecticides a entraîné une augmentation due 2,8 fois de la DL 50 au diflubenzuron et de 3,9 fois au méthoprène. Comparés aux souches standards sensibles de Colombo, les taux de résistance des lignées de Dar-es-Salam se sont élevés de 2,4 à 6,6 pour le diflubenzuron et de 6,1 à 21,3 pour le méthoprène, sans effet visible significatif sur la résistance croisée. Ces résultats sont discutés en relation avec la standardisation du test IGR et les besoins de programmes de lutte intégrée contre C. quinquefasciatus.

     

Changes in malathion and pyrethroid resistance after cypermethrin selection of Culex quinquefasciatus field populations of Cuba

Abstract. Use. of the organophosphorus insecticide malathion for mosquito control in Cuba, for 7 years up to 1986, selected elevated non-specific esterase and altered acetylcholinesterase (AChE) resistance mechanisms in Culex quinquefasciatus. In central Havana space-spraying of malathion was replaced by the pyrethroid cypermethrin in 1987: alternate cycles of malathion and cypermethrin were applied in some of the more rural areas of Havana district during 1987-91. Consequently, populations of Cx quinquefasciatus in the central area of Havana developed resistance to cypermethrin, but there is no evidence of pyrethroid resistance in the outlying areas. Malathion resistance levels declined significantly after 1986, measured both by bioassay and the frequency of the elevated esterase resistance mechanism, and then stabilized with no measurable decline during 1990 in any of the populations tested. These populations had less than 10% frequency of susceptible homozygotes for both the esterase and AChE resistance mechanisms, indicating that organophosphate resistance is still prevalent in Cuban Cx quinquefasciatus. These two mechanisms appear to be in linkage equilibrium, suggesting that current selection for double resistance is not strong. In the central Havana region, pirimiphos-methyl, an organophosphorus insecticide unaffected by the two common malathion resistance mechanisms, is now being used in a resistance management strategy designed to avoid pyrethroid resistance spreading.     

Overwintering of Culex mosquitoes in Sweden and their potential as reservoirs of human pathogens

1. Culex pipiens L. and Cx torrentium Martini are suspected enzootic vectors in Sweden of Sindbis virus causing Ockelbo disease in man. To estimate their potential as virus reservoirs over the winter, mosquitoes were collected from August to May from a cellar in a rural area, central Sweden. The mosquitoes were dissected and examined for blood content, gonotrophic stage, parity and insemination, and tested for fructose. 2. Cx pipiens, Cx torrentium and Cx territans Walker constituted 94% of 887 females caught. A few Culiseta annulata (Schrank), Cs. alaskaensis (Ludlow) and An. maculipennis Meigen s.l. were also collected. 3. Data suggested that all blood-fed, parous Culex females disappeared during the winter and that only non-bloodfed, nulliparous females survived. 4. Fructose, detected in some of these females, and other plant sugars may be the main energy sources for winter survival. 5. Unless transovarially or venerally infected, Culex females are presumably of no or very limited importance in rural, central Sweden as winter reservoirs for blood-borne pathogens of vertebrates.     

Autumn southward ‘return’ migration of the mosquito Culex tritaeniorhynchus in China

Abstract. Direct evidence for a southward ‘return’ migration in autumn of the mosquito Culex tritaeniorhynchus in China was obtained by aerial sampling with a net suspended from a balloon. In a preliminary study between 2 and 20 September 1990 at Jiangpu, near Nanjing, in Jiangsu Province, 11 females and 4 males of this species were taken at heights of 150–250 m. During 17–26 October 1991 at Dongxiang in northern Jiangxi Province, 44 mosquitoes (8 males, 36 females) were caught in the aerial net at heights of 80–380m in northeasterly winds (E-NNW). Most of the specimens were flying within the subsiding air-mass behind a cold front. Cx tritaeniorhynchus was the only species identified (31 females) among the mosquitoes from Dongxiang. Of 24 females dissected, 17 had N stage ovaries — interpreted as diapause, five had stage I ovaries, one had stage II ovaries, and one was gravid (stage V), but none was freshly bloodfed. Cx tritaeniorhynchus is the main vector of Japanese viral encephalitis in China, and it is possible that the virus is reintroduced to northern temperate areas in spring by northward migration of infected Cx tritaeniorhynchus females.     

Purification and characterization of mosquitocidal Bacillus sphaericus BinA protein

Certain strains of Bacillus sphaericus produce a highly toxic mosquito-larvicidal binary toxin during sporulation. The binary toxin is composed of toxic BinA (41.9 kDa) and receptor binding BinB (51.4 kDa) polypeptides and is active against vectors of filariasis, encephalitis and malaria. The toxin has been tested with limited use for the control of vector mosquitoes for more than two decades. The binA gene from a local ISPC-8 strain of B. sphaericus that is highly toxic to Culex and Anopheles mosquito species was cloned into pET16b and expressed in Escherichia coli. The purified BinA protein differs by one amino acid (R197 M) from BinA of the highest toxicity strains 1593/2362/C3-41. Majority of the expressed protein was observed in inclusion bodies. BinA inclusions alone from E. coli did not show toxic activity, like reported previously. However, the active form of BinA could be purified to homogeneity from the soluble fraction of E. coli cell lysate, grown at reduced temperature after isopropyl β-d-thiogalactopyranoside induction. The purified BinA protein with and without poly-histidine tag showed LC₅₀ dose of 82.3 and 66.9 ng ml⁻¹, respectively, at 48 h against Culex quinquefasciatus larvae. The secondary structure of BinA is expected to be mainly β strands as estimated using far-UV circular dichroism. The estimates matched well with the secondary structure predictions using amino acid sequence. This is the first report of large-scale purification and accurate toxicity estimation of soluble B. sphaericus BinA. This can help in design and synthesis of improved bacterial insecticide.