Laboratory evaluation of Mesocyclops annulatus (Wierzejski, 1892) (Copepoda: Cyclopidea) as a predator of container-breeding mosquitoes in Argentina

In laboratory bioassays we tested the predatory capacity of the copepod Mesocyclops annulatus on Aedes aegypti and Culex pipiens larvae. A single adult female of M. annulatus caused 51.6% and 52.3% mortality of 50 first instar larvae of Ae. aegypti and Cx. pipiens respectively, in a 72 h test period. When alternative food was added to the containers, mortality rates declined to 16% and 10.3% for Ae. aegypti and Cx. pipiens respectively. When 50 first instar larvae of each of the two mosquito species tested were placed together with a single adult female of M. annulatus, mortality rates were 75.5% for Ae. aegypti larvae and 23.5% for Cx. pipiens larvae in a three day test period. Different density of adult females of M. annulatus ranged from 5 to 25 females produced mortality rates of Ae. aegypti first instar larvae from 50% to 100% respectively. When a single adult female of M. annulatus was exposed to an increasing number of first-instar Ae. aegypti larvae ranging from 10 to 100, 100% mortality was recorded from 1 to 25 larvae, then mortality declined to 30% with 100 larvae. The average larvae killed per 24 h period by a single copepod were 29.     

竹叶花椒果实精油对两种蚊虫的毒杀活性研究

采用浸液法测试了竹叶花椒果实的水蒸汽蒸馏精油对白纹伊蚊和致倦库蚊幼虫的毒杀效果,并用三角瓶密闭熏蒸法研究了精油对这两种成蚊的熏杀活性;此外,采用GC-MS分析了该精油的化学成分。研究结果:(1)精油对白纹伊蚊和致倦库蚊的Ⅰ、Ⅱ、Ⅲ、Ⅳ龄期幼虫及蛹的LC50值分别为25.634/61.472、31.763/76.431、52.356/110.172、258.497/121.884和198.263/162.048mg.L-1;(2)精油对白纹伊蚊和致倦库蚊成蚊的LC50值分别为24.957和29.517μg.cm-3;(3)在147.52μg.cm-3熏杀剂量下,精油对白纹伊蚊和致倦库蚊成蚊的KT50值分别为3.493和2.993min,24h致死率均为100%;(4)共鉴定出18种化合物,其中萜烯类10种,占精油总量的67.122%,为竹叶花椒果实精油的主要成分。竹叶花椒果实精油对白纹伊蚊和致倦库蚊均有明显的致死作用,且作用速度快,具有开发为植物源杀蚊剂的潜力。     

Ticks and mosquitoes as vectors of Borrelia burgdorferi s. l. in the forested areas of Szczecin

The aim of the study was to determine the infection level of adult forms and larvae of ticks and mosquitoes with Borrelia burgdorferi in the forested areas of Szczecin. A total of 1699 ticks Ixodes ricinus, including 1422 nymphs, 277 adult forms and 2862 mosquito females representing the genera Aedes (89.6%) and Culex (10.4%) were collected between the years 2004 and 2005. A further 3746 larvae and 1596 pupae of Culex pipiens pipiens were colleted from water bodies. Borrelia burgdorferi s. l. was detected in the arthropods by the method of indirect immunofluorescence assay (IFA). A positive immunological reaction was detected in 16.6% of the adult forms and in 16.5% of the nymphs of Ixodes ricinus. Spirochetes were also detected in 1.7% of mosquito females, 3.2% of larvae and in 1.6% of pupae of Culex pipiens pipiens. The results of the present study confirm that contact with ticks constitutes the main risk of contracting Lyme disease, although mosquitoes play a role as vectors as well.     

小蓬草精油对两种蚊虫的毒杀活性和成分分析

【目的】明确小蓬草Conyza canadensis ( L.) Cronq.精油的杀虫潜力及其活性成分。【方法】通过浸虫法和密闭熏蒸法测试了小蓬草精油对白纹伊蚊Aedes albopictus和致倦库蚊Culex pipiens quinquefasciatus幼虫及成蚊的毒杀活性, 并利用气相色谱 质谱联用仪（GC-MS）对精油的挥发性成分进行了定性分析。【结果】 小蓬草精油对白纹伊蚊1-4龄期幼虫及蛹的24 h LC50值分别为25.01, 45.88, 56.94, 64.60和346.23 μg/mL; 对致倦库蚊幼虫1-4龄期幼虫及蛹的24 h LC₅₀值分别为9.16, 8.65, 32.12, 43.68和197.83 μg/mL。在剂量分别为48, 64, 80, 96, 112和128 μg/cm3时, 小蓬草精油对白纹伊蚊成蚊的KT₅₀值分别为28.81, 22.31, 20.38, 17.05, 13.92和9.74 min; 对致倦库蚊的KT50值分别为34.90, 32.97, 23.97, 19.60, 15.20和10.34 min。 24 h熏蒸对白纹伊蚊和致倦库蚊成蚊的LC50值分别是75.46和99.19 μg/cm³。小蓬草精油的GC MS定性分析共分离鉴定出31种化合物, 其中单萜类物质6种, 倍半萜烯类物质17种, 含氧萜烯类6种。【结论】结果表明小蓬草精油对这两种蚊虫的毒杀活性较高, 具有深开发潜力。     

椿叶花椒果实精油对两种蚊虫的生物活性及成分分析

为筛选环保型的植物源灭蚊活性物质, 研究了椿叶花椒 Zanthoxylum ailanthoides 果实精油对白纹伊蚊Aedes albopictus和致倦库蚊Culex pipiens quinquefasciatus成蚊的熏杀活性, 并测试了精油对这两种蚊幼虫的毒杀效果。此外, 采用GC-MS分析了该精油的化学成分。 结果表明：在102.145 μg·cm^-3熏杀剂量下, 精油对白纹伊蚊和致倦库蚊成蚊的KT50值分别为2.788和3.505 min, 24 h致死率分别为100%和97.53%。该精油对白纹伊蚊和致倦库蚊4龄幼虫的LC50值分别为45.237和20.759 mg·L^-1。从椿叶花椒果实精油中共鉴定出14种化合物, 其中2-十一酮为主要成分, 相对含量为89.86%。结果说明椿叶花椒果实精油对两种蚊虫均有明显的杀虫活性, 具有开发成植物源灭蚊剂的潜力。     

Effects of temperature, pH and salinity on the infection of Leptolegnia chapmanii Seymour (Peronosporomycetes) in mosquito larvae

The effects of temperature, pH, and NaCl concentrations on the infectivity of zoospores of Leptolegnia chapmanii (Argentine isolate) were determined for Aedes aegypti and Culex pipiens under laboratory conditions. Zoospores of L. chapmanii were infectious at temperatures between 10 and 35 degrees C but not at 5 or 40 degrees C. At the permissive temperatures, mortality rates in young instars were much higher than in older instars and larvae of Ae. aegypti were more susceptible to L. chapmanii than larvae of Cx. pipiens. At 25 degrees C, Ae. aegypti larvae challenged with L. chapmanii zoospores resulted in 100% infection at pH levels ranging from 4 to 10. Larvae of Cx. pipiens exposed to similar pH and zoospore concentrations resulted in increasing mortality rates from 62% to 99% at pH 4 to 7, respectively, and then decreased to 71% at pH 10. Aedes aegypti larvae exposed to L. chapmanii zoospores in NaCl concentrations ranging from 0 to 7 parts per thousand (ppt) at 25 degrees C resulted in 100% mortality while mortality rates for Cx. pipiens decreases from 96% in distilled water to 31.5% in water with 6 ppt NaCl. Control Cx. pipiens larvae died when exposed at a NaCl concentration of 7 ppt. Vegetative growth of L. chapmanii was negatively affected by NaCl concentrations. These results have demonstrated that the Argentinean isolate of L. chapmanii tolerated a wide range of temperatures, pH, and salinity, suggesting that it has the potential to adapt to a wide variety of mosquito habitats.     

用刘氏中剑水蚤防治蚊幼虫(英文)

本研究在室内评价了在济宁采集的捕食性刘氏中剑水蚤单用及与球形芽孢杆菌(B.s.)或苏云金杆菌以色列变种(B.t.i.)合用对蚊幼虫的控制效果。在室内,刘氏中剑水蚤对白蚊伊蚊Ⅰ龄幼虫的捕食效果好于对中华按蚊和淡色库蚊幼虫的效果。蚊笼模拟试验表明,刘氏中剑水蚤可分别于9、10和7周内消灭淡色库蚊、中华按蚊和白蚊伊蚊的幼虫种群。该剑水蚤增长速率快,大量培养经济易行。现场试验证明,刘氏中剑水蚤虽可在蚊虫孳生地存活时间长于蚊虫季节,但单用该剑水蚤不能象化学杀虫剂一样快速完全地控制三种蚊虫。刘氏中剑水蚤与B.s.或B.t.i.合用,应用刘氏中剑水蚤的长持效和生物杀虫剂的速效,取得了高效、持久的防治效果。因此二者合用可用于蚊虫防治并可作为蚊虫综合防治的一个组成部分。     

BIOLOGICAL CONTROL OF MOSQUITO LARVAE WITH MESOCYCLOPS LEUKARTI1)

Abstract  The control efficacy of a predatory copepod, Mesocyclops leukarti , collected in Jining, and its compatibility with Bacillus sphaericus (B.s. ) and B. thuringiensis var. csraefensis H-14 ( B. t. i. ) against mosquito larvae were evaluated in the laboratory and in the field. In the laboratory, M. leukarti showed higher predatory activity toward first instar larvae of Aedes albopictus than toward that of Anopheles sinensis and Culex pipiens pallens. Cage simulation test showed that larvae of Cx. pipiens pallens, An. sinensis and Ae. albopictus could be eliminated within 9, 10 and 7 weeks after interaction with the copepods respectively. M. leukarti population can grow quickly, and its mass production is easy and economic. However, pilot field trials indicated that M. leukarti alone could not eradicate mosquito larvae of the three species rapidly and completely as by chemical insecticides, although they could persist in mosquito breeding habitats longer than the mosquito season. When Mesocyclops was used in combination with B. s. and B. t. i. , higher and sustainable control effects were achieved. So this combination, which achieved the rapid larvicidal and sustainable control effect, was presumed to be practical and could be integrated into mosquito control program.     

Isolation and identification of α-proteobacteria from Culex pipiens (Diptera Culicidae) larvae

A survey of drainage ditches in suburban areas of La Plata, Buenos Aires province, Argentina for pathogens of Culex pipiens larvae was conducted from 2003 to 2006. C. pipiens larvae of opaque, white color were found in several of those field collections. When the white larvae were dissected and observed by phase-contrast microscopy in wet-mount preparations, the presence of bacteria, located in the hemocoel, was recorded. Laboratory experiments were performed to elucidate the pathway for transmission of this pathogen. Although approaches involving traditional culturing had failed to reveal the identity of the new microorganism present, molecular techniques to identify the pathogen in the studies reported here were successful. The partial sequence of the 16S-rRNA gene constitutes a powerful tool for the detection of new isolates from the hemocoele of C. pipiens larvae. These bacteria were characterized as belonging to the genus Novispirillum. In spite of the genus's wide distribution in different aquatic environments, information related to the parasitic relationship of Novispirillum spp. to aquatic insects is scarce, and this association has not been described in other mosquito species. This report constitutes the first documentation of Novispirillum spp. as a pathogen for mosquito larvae.     

Cloning and overexpression of CYP6F1, a cytochrome P450 gene, from deltamethrin-resistant Culex pipiens pallens

CYP6F1 (GenBank/EMBL accession No. AY662654), a novel gene with a complete encoding sequence in the cytochrome P450 family 6, was cloned and sequenced from deltamethrin-resistant 4th instar larvae of Culex pipiens pallens. The cDNA sequence of CYP6F1 has an open reading frame of 1527 bp, which encodes a putative protein of 508 amino acid residues. The deduced amino acid sequence of CYP6F1 indicated that the encoded P450 has conserved domains of a putative membrane-anchoring signal,putative reductase-binding sites, a typical heme-binding site, an ETLR motif and substrate recognition sites.Semi-quantitative RT-PCR analysis indicated that the CYP6F1 gene was expressed to a greater extent in the deltamethrin-resistant strain than in the susceptible strain of Cx. pipiens pallens. The expression levels of the CYP6F1 gene in the deltamethrin-resistant 1 st, 2nd, 3rd, 4th instar larvae and adult female mosquitoes differed, with highest expression levels in the 4th instar larvae. In addition, the CYP6F1 gene was stably expressed in mosquito C6/36 cells, and the expected 61.2 kDa band was identified by Western blotting. The cells transfected with CYP6F1 had an increased resistance to deltamethrin as compared with control cells.These results indicate that CYP6F1 is expressed at higher levels in the deltamethrin-resistant strain, and may confer some insecticide resistance in Cx. pipiens pallens.     

Characterization of phospholipase B of Culex pipiens fatigans

Phospholipase B has been found in the mosquito Culex pipiens fatigans, and some of its properties have been studied. The enzyme had a high optimum temperature (45 degrees C) and broad alkaline pH optimum (8-9). It was inactive toward diacylphospholipids, and hydrolyzed lysolecithin at a higher rate than lysophosphatidyl ethanolamine. The enzyme was heat labile, but lysolecithin protected it against heat inactivation. Phosphatidyl ethanolamine, phosphatidyl choline, deoxycholate, Fe(+++), and Hg(++) inhibited the enzyme markedly. The enzyme was present mainly in larvae; little enzyme activity was detected in pupae or adults. The total and specific activities were highest in IV instar (6 day) and I instar (1st day) larvae, respectively. It was localized in the microsomal fraction and was distributed mainly in the abdomen and thorax of the insect. The enzyme was present at much higher levels of activity in larvae of the mosquitoes Anopheles stephensi and Aedes aegypti.     

Host range and specificity of an Argentinean isolate of the aquatic fungus <Emphasis Type="Italic">Leptolegnia chapmanii</Emphasis> (Oomycetes: Saprolegniales), a pathogen of mosquito larvae (Diptera: Culicidae)

An isolate from Argentina of the fungal mosquito pathogen Leptolegnia chapmanii (ARSEF 5499), was tested against 12 species of mosquito larvae and on species of non-target aquatic invertebrates and vertebrates. The mosquito species tested were Aedes aegypti, Anopheles sp., Culex apicinus, Cx. castroi, Cx. dolosus, Cx. pipiens, Cx. renatoi, Isostomyia paranensis, Ochlerotatus albifasciatus, Oc. crinifer, Psorophora cyanescens, and P. ferox. Mosquito larvae of 10 species were susceptible, with mortality rates from 10–100%. Two mosquito species Cx. renatoi and I. paranensis were not infected by Leptolegnia. None of the non-target fauna treated was infected by L. chapmanii with exception of members of the Family Chironomidae which were susceptible at low infection rates. Researcher CIC (Committee for Scientific Research of Buenos Aires State).     

Oxygen consumption in mosquito larvae parasitized by Romanomermis culicivorax (Nematoda)

The effect of parasitism by Romanomermis culicivorax on the oxygen consumption of three species of mosquito larvae was studied by manometric methods. The Q02 of Culex pipiens, both parasitized and unparasitized, decreased as the host larvae developed from the second to fourth instar. In the early stages there was no significant difference in Q02 in parasitized and unparasitized hosts but the Q02 increased significantly in parasitized fourth instar larvae. The Q02 of parasitized and unparasitized fourth instars of Aedes taeniorhynchus, Anopheles quadrimaculatus and C. pipiens were compared. The Q02 of all parasitized hosts was significantly increased and the Q02 of the two culicine species displayed a dramatic increase in oxygen consumption. The oxygen consumption of the parasitic mermithid larvae was estimated from that obtained earlier for the post-parasitic larvae but did not account for more than 9-36% of the increased Q02 of the parasitized hosts. On this basis it seems probable that the increased metabolic rate as reflected by oxygen consumption of the parasitized mosquito larve was principally a manifestation of the physiological and nutritional stress induced by the mermithid nematodes.     

Host range of the parasite Strelkovimermis spiculatus (Nematoda: Mermithidae) in Argentina mosquitoes

Strelkovimermis spiculatus ( Poinar and Camino 1986 ) is a nematode parasite of mosquitoes isolated from the Neotropical region. We investigated the host range of this parasite in mosquito populations for a better understanding of the dynamics of temporary breeding sites. Five grassy‐pool habitats filled by rainwater were sampled from the summer 2007 through the fall 2008. Eight mosquito species were collected: Anopheles albitarsis, Culex chidesteri, Culex dolosus, Culex maxi, Aedes albifasciatus, Psorophora ciliata, Psorophora cyanescens, and Psorophora albigenu. Six of these species were parasitized: Cx. chidesteri, Cx. dolosus, Cx. maxi, Ae. albifasciatus, Ps. ciliata, and Ps. cyanescens. The occurrence of this mermithid in natural mosquito populations was increased from the end of winter to the end of the spring. Prevalence ranged from 11% to 100%. High levels of infections were registered only in Ae. albifasciatus larvae, the most abundant mosquito species (95%), followed by Cx. dolosus (2.7%). Strelkovimermis spiculatus completed its development in all infected mosquito larvae. The presence of S. spiculatus in six natural mosquito populations increases the number of susceptible species to 24.     

Results of a survey to detect the mosquito Aedes albopictus in the French Riviera.

A programme of surveillance was initiated in 1992, in the French Riviera, to detect a possible introduction of Aedes albopictus from Italy and to prevent nuisances caused by mosquitoes in the touristic localities of the C?te d'Azur. In five years, numerous mosquito breeding places have been located. Nine species have been collected: Anopheles claviger, An. plumbeus, Aedes geniculatus, Ae. vittatus, Culex hortensis, Cx. impudicus, Cx. pipiens, Culiseta fumipennis, Cs. longiareolata but no Ae. albopictus was found. Nuisances were mainly due to hypogean populations of Cx. pipiens. Breeding places in urban sites have been controlled or suppressed. The discovery of an important larval population of An. plumbeus in an urban area might further stress the importance of this species already suspected to transmit indigenous malaria in cities.     

Evaluation of some adhesives for collecting Musca domestica and Chrysomya megacephala adults or mosquito larvae in sticky traps

1. Seven types of water-insoluble adhesives were evaluated in sticky traps for collecting adults of Musca domestica L. and Chrysomya megacephala (Fabricius) or mosquito larvae (Aedes aegypti (L.) and Culex quinquefasciatus Say). 2. Adhesive viscosity affected the tackiness of the glues and this determined their trapping efficiency in air or water. 3. From the 'Hyvis' range of adhesives tested, 'Hyvis 200' was most effective for trapping adult flies. 4. With 24 h exposure to fourth instar Ae.aegypti larvae in tapwater, submerged plates coated with 'Hyvis 10', 'Hyvis 30' or 'Hyvis 200' formulations trapped the majority of larvae. In polluted water the highest rates of trapping were 17.3% of Ae.aegypti and 18.7% of Cx quinquefasciatus with 'Hyvis 200'. Floating traps were consistently less productive than submerged traps under laboratory conditions. 5. In a heavily polluted natural breeding-site of Cx quinquefasciatus, floating traps were more productive than submerged sticky traps with four of seven adhesives tested, the most efficient being 'Hyvis 200' (4.2 mosquitoes per hour) and Hyvis:polyethylene 90:10 (4.5/h). Despite the relative inefficiency of aquatic traps, emergent adults, pupae and second to fourth instars of larvae were collected quickly from the habitat.     

Evaluation of various control agents against mosquito larvae in rice paddies in Taiwan.

A field test was conducted in rice paddies adjacent to Wufeng, Taichung County in Central Taiwan to evaluate the efficacy of control agents against mosquito larvae. The agents included Bacillus thuringienesis israelensis (Bti), two Lagenidium giganteum products (Lg product A and T), and temephos. The major mosquito species found in the rice paddies were Culex tritaeniorhynchus/vishnui and Anopheles sinensis. Compared to controls, a 7-day treatment with Bti or Lg products A and T caused overall reductions in the number of immatures (larvae and pupae) of Cx. tritaeniorhynchus/vishnui of 77.5%, 49.7%, and 21.9%, respectively, whereas temephos caused an increase of 66.9%. The overall reductions in An. sinensis were 85.4%, 8.6%, 44.6%, and 92.1%, respectively. There was no significant reduction in the number of mosquito larvae following 42 days of treatment with these agents. In summary, 1-week treatments with both biological control agents produced moderate overall reductions in mosquito larvae in rice paddies. The insecticide temephos, on the other hand, was very effective at suppressing the larvae of An. sinensis but significantly increased the number of Cx. tritaeniorhynchus/vishnui larvae in temephos-treated plots.     

The taxonomy and biology of two sympatric sibling species of Culex, C. pipiens and C. torrentium (Diptera, Culicidae)

The mosquito Culex torrentium Martini is a sibling species of C. pipiens L. and was first discovered in Britain in 1951. However, it is now evident that it was present in this country from at least 1900. Collections from a few southern localities in England indicate that the size of its population is about a third that of C. pipiens.
A comparative taxonomic study showed that neither the eggs nor larvae can be separated from those of C. pipiens , and that only in certain instances can the pupae of the two species be distinguished. Males are readily separated on the structure of the terminalia. Previously it was considered that C. torrentium could be distinguished by the presence of a pre-alar patch of scales, but it is shown that in a few specimens they are absent and in others they may become easily detached.
Larvae of both species frequently occur together in the same habitats, and no differences were observed in the biology of their immature stages. Serological tests showed that adults of both species feed on birds. Adult females of both species hibernate during the autumn, but whereas those of C. pipiens are common in man-made shelters, C. torrentium is absent.     

Midgut barrier imparts selective resistance to filarial worm infection in Culex pipiens pipiens

Mosquitoes in the Culex pipiens complex thrive in temperate and tropical regions worldwide, and serve as efficient vectors of Bancroftian lymphatic filariasis (LF) caused by Wuchereria bancrofti in Asia, Africa, the West Indies, South America, and Micronesia. However, members of this mosquito complex do not act as natural vectors for Brugian LF caused by Brugia malayi, or for the cat parasite B. pahangi, despite their presence in South Asia where these parasites are endemic. Previous work with the Iowa strain of Culex pipiens pipiens demonstrates that it is equally susceptible to W. bancrofti as is the natural Cx. p. pipiens vector in the Nile Delta, however it is refractory to infection with Brugia spp. Here we report that the infectivity barrier for Brugia spp. in Cx. p. pipiens is the mosquito midgut, which inflicts internal and lethal damage to ingested microfilariae. Following per os Brugia exposures, the prevalence of infection is significantly lower in Cx. p. pipiens compared to susceptible mosquito controls, and differs between parasite species with <50% and <5% of Cx. p. pipiens becoming infected with B. pahangi and B. malayi, respectively. When Brugia spp. mf were inoculated intrathoracically to bypass the midgut, larvae developed equally well as in controls, indicating that, beyond the midgut, Cx. p. pipiens is physiologically compatible with Brugia spp. Mf isolated from Cx. p. pipiens midguts exhibited compromised motility, and unlike mf derived from blood or isolated from the midguts of Ae. aegypti, failed to develop when inoculated intrathoracically into susceptible mosquitoes. Together these data strongly support the role of the midgut as the primary infection barrier for Brugia spp. in Cx. p. pipiens. Examination of parasites recovered from the Cx. p. pipiens midgut by vital staining, and those exsheathed with papain, suggest that the damage inflicted by the midgut is subcuticular and disrupts internal tissues. Microscopic studies of these worms reveal compromised motility and sharp bends in the body; and ultrastructurally the presence of many fluid or carbohydrate-filled vacuoles in the hypodermis, body wall, and nuclear column. Incubation of Brugia mf with Cx. p. pipiens midgut extracts produces similar internal damage phenotypes; indicating that the Cx. p. pipiens midgut factor(s) that damage mf in vivo are soluble and stable in physiological buffer, and inflict damage on mf in vitro.