Taxonomic studies on Culex (Melanoconion) coppenamensis Bonne-Wepster & Bonne (Diptera: Culicidae), and description of two new species from Brazil

Two new species,Culex (Melanoconion) alinkios and Culex (Melanoconion) symbletos are described and defined based on morphological features of the male genitalia. The former is from Vale do Ribeira, Atlantic Forest, southeastern of the state of S?o Paulo, Brazil, and belongs to the Bastagarius subgroup. The latter is from the Parque Nacional do Jaú, state of Amazonas, Brazil, and belongs to the Inhibitator subgroup and is similar to Cx. mesodenticulatus Galindo and Mendez. Diagnostic characters for the identification of the adult males of the species are provided. Two morphological forms (Form 1 and 2), which are similar to Cx. coppenamensis, were also found in the Parque Nacional do Jaú. Form 1 is described and compared with the new species from Vale do Ribeira and Form 2.     

Diel biting activity of Culex (Melanoconion) caudelli in Trinidad, West Indies

The diel biting periodicity of the arbovirus vector Culex (Melanoconion) caudelli Dyar and Knab (Diptera; Culicidae) in the Aripo-Wallerfield forest, Trinidad, was studied by collecting mosquitoes attracted to mouse-baited traps at 2-hourly intervals during eight 24 h periods. Biting females of Cx caudelli were collected during all night-time hours, 18.00-06.00 hours, with an overall unimodal pattern, i.e. one well-defined peak between 22.00 and 04.00 hours. This contrasts with previous reports that Cx caudelli is diurnally active. During the four moon phases, the period of peak biting activity varied from 22.00 to 24.00 hours at fullmoon, from 22.00 to 04.00 hours at first quarter, and from 24.00 to 02.00 hours at new moon and last quarter phases. The number of mosquitoes collected varied significantly with moon phases, the highest (152/24 h) being collected in the first lunar quarter and the lowest (71/24 h) in the last lunar quarter. Cloud cover and rainfall had no significant effect on the diel biting periodicity of Cx caudelli.     

Distribution and seasonality of Culex (Melanoconion) caudelli, a vector of arboviruses in Trinidad

Seasonal and spatial distributions of the arbovirus vector mosquito Culex (Melanoconion) caudelli Dyar and Knab (Diptera: Culicidae) in the Aripo-Wallerfield forest, Trinidad, were studied by collecting mosquitoes attracted to mouse-baited traps in forest, ecotone and savannah, one day per week throughout the year, 98% of the Cx caudelli mosquitoes were collected during the wet season, with a significant positive correlation between rainfall and mosquito catch. The reverse occurred during the dry season. Cx caudelli displayed a preference for the ecotone where 67% were collected, while 32% and 1% were collected in the forest and savannah respectively.     

The arbovirus vector Culex torrentium is more prevalent than Culex pipiens in northern and central Europe

Two species of arbovirus vector, Culex torrentium and Culex pipiens (Diptera: Culicidae), occur in several European countries, but difficulties in their accurate identification and discrimination have hampered both detailed and large‐scale distribution and abundance studies. Using a molecular identification method, we identified to species 2559 larvae of Cx. pipiens/torrentium collected from 138 sites in 13 European countries ranging from Scandinavia to the Mediterranean coast. In addition, samples of 1712 males of Cx. pipiens/torrentium collected at several sites in the Czech Republic were identified to species based on the morphology of their hypopygia. We found that the two species occur together in large areas of Europe, and that Cx. torrentium dominates in northern Europe and Cx. pipiens dominates south of the Alps. The transition in dominance occurs in central Europe, where both species are roughly equally common. There was a strong correlation between the length of the growing season at different sites and occurrences of the two species. As the growing season increases, the proportion and detection of Cx. torrentium decrease, whereas those of Cx. pipiens increase. The present findings have important consequences for the interpretation of the results of studies on major enzootic and link‐vectors of mosquito‐borne bird‐associated viruses (i.e. Sindbis, West Nile and Usutu viruses), especially in central Europe and Scandinavia.     

The use of morphometric wing characters to discriminate female Culex pipiens and Culex torrentium

The reliability of the length of wing radial vein r_2/3 as a character for the morphological discrimination of the two potential arbovirus vectors Culex pipiens s.s. and Cx. torrentium from Germany was reassessed, after this character had been neglected for more than 40 years. Additionally, multivariate morphometric analyses were applied to evaluate wing shape variation between both species. Although high‐throughput molecular tools are now available to differentiate the two species, a simple, low‐cost routine alternative may be useful in the absence of a molecular laboratory, such as under semi‐field conditions. A thin‐plate splines transformation confirmed that primarily the shrinkage of vein r_2/3 is responsible for the wing differences between the two species. In the bivariate analysis, the r_2/3/r₃ indices of Cx. pipiens s.s. and Cx. torrentium were 0.185 and 0.289, respectively, resulting in a correct classification of more than 91% of all tested specimens. Using the absolute length of vein r_2/3 alone still allowed for more than 90% accurate discrimination. Furthermore, classification accuracy of linear discriminant analysis exceeded 97%.     

A sensitive and reliable restriction enzyme assay to distinguish between the mosquitoes Culex torrentium and Culex pipiens

Culex pipiens pipiens Linnaeus and Culex torrentium Martini (Diptera: Culicidae) are closely related vector species that exist sympatrically in Europe. The two species are morphologically almost identical and can only be distinguished with certainty by characters of the male genitalia. Hence, correct species identification and conclusions on distribution and vector status are very difficult and often neglected. Therefore, we developed a reliable and simple mitochondrial cytochrome c oxidase subunit I (COI) gene restriction enzyme assay to discriminate between Cx. pipiens and Cx. torrentium, based on the analysis of morphologically identified male specimens. We sequenced approximately 830 bp in the 3′ region of the mitochondrial COI gene of 18 morphologically identified males of Cx. pipiens and Cx. torrentium. Two restriction enzymes (FspBI and SspI) that could distinguish between the two species according to species‐specific differences in these sequences were chosen. The restriction enzymes were tested on 227 samples from Sweden and verified by sequencing 44 of them. The enzyme FspBI correctly identified all investigated samples; the enzyme SspI identified all samples except one Cx. torrentium. We hope the method and the findings presented here will help to shed light on the true distribution and relative proportions of the two species in Europe.     

Wide recognition of Culex pipiens and lack of detection of Culex torrentium through biomolecular differentiation of mosquitoes in the Emilia‐Romagna region,Northern Italy

The Culex pipiens complex includes species with reported differences in vector competence for arthropod‐borne viruses, many of which are of significant importance to human health such as the West Nile virus and the Sindbis virus. This group of mosquitoes is difficult to distinguish morphologically; particularly as adult females. In Europe, the two species of the complex, Culex pipiens Linnaeus 1758 and Culex torrentium Martini 1925, are often found sympatrically. With the aim to characterize the presence and spread of both species in the Emilia‐Romagna region, Northern Italy, mosquitoes of the complex – collected during the West Nile virus surveillance plans – were tested by multiplex real‐time PCR for the detection of the two species Cx. pipiens and Cx. torrentium. A total of 24 165 mosquitoes, collected between 2012 and 2014 from 105 sites, and sorted in 204 pools, were tested. All tested pools were found to be composed of Cx. pipiens, whereas Cx. torrentium was not detected. These results indicate a likely absence of Cx. torrentium mosquitoes within the surveyed territory, whereas Cx. pipiens is widely distributed in the area mentioned. This is in line with previous reports, which describe a pre‐alpine distribution of Cx. torrentium in Italy.     

Population Structure and Distribution Patterns of the Sibling Mosquito Species Culex pipiens and Culex torrentium (Diptera: Culicidae) Reveal Different Evolutionary Paths

Nowadays a number of endemic mosquito species are known to possess vector abilities for various diseases, as e.g. the sibling species Culex pipiens and Culex torrentium. Due to their morphological similarity, ecology, distribution and vector abilities, knowledge about these species'' population structure is essential. Culicidae from 25 different sampling sites were collected from March till October 2012. All analyses were performed with aligned cox1 sequences with a total length of 658 bp. Population structure as well as distribution patterns of both species were analysed using molecular methods and different statistical tests like distance based redundancy analysis (dbDRA), analysis of molecular variances (AMOVA) or McDonald & Kreitman test and Tajima''s D. Within both species, we could show a genetic variability among the cox1 fragment. The construction of haplotype networks revealed one dominating haplotype for Cx. pipiens, widely distributed within Germany and a more homogeneous pattern for Cx. torrentium. The low genetic differences within Cx. pipiens could be a result of an infection with Wolbachia which can induce a sweep through populations by passively taking the also maternally inherited mtDNA through the population, thereby reducing the mitochondrial diversity as an outcome of reproductive incompatibility. Pairwise population genetic differentiation (F_ST) ranged significantly from moderate to very great between populations of Cx. pipiens and Cx. torrentium. Analyses of molecular variances revealed for both species that the main genetic variability exists within the populations (Cx. pipiens [88.38%]; Cx. torrentium [66.54%]). Based on a distance based redundancy analysis geographical origin explained a small but significant part of the species'' genetic variation. Overall, the results confirm that Cx. pipiens and Cx. torrentium underlie different factors regarding their mitochondrial differentiation, which could be a result of endosymbiosis, dispersal between nearly located populations or human introduction.     

Differential effects of Bacillus sphaericus strain 2362 on Culex quinquefasciatus and its competitor Culex cinereus in West Africa

The larval susceptibility to Bacillus sphaericus strain 2362 of the non-man-biting mosquito Culex cinereus and the urban filariasis vector Culex quinquefasciatus, two competitor mosquitoes in polluted habitats, was compared. In the laboratory, both species ingested a similar amount of B. sphaericus spores when fed c. 2 x 10(5) spores per ml for 30 min. However, in the same experiment, third-instar larvae of Cx quinquefasciatus were reduced by 98% at 24 h exposure while Cx cinereus larvae were only reduced by 6% at 72 h. In the field, preimaginal populations of Cx cinereus ingested, within a week, more than 99% of the applied spores, but showed no significant decrease through 14 days in cesspools treated at 10 g/m2 of a flowable concentrate of B. sphaericus 2362, containing 2 x 10(10) spores/g. It is proposed that specific biological control of Cx quinquefasciatus could result from appropriate treatment of breeding-sites with larvicidal B. sphaericus and competitive displacement by Cx cinereus or other mosquitoes with larvae that are more tolerant of B. sphaericus.     

Butterflies of the South Pacific

ABSTRACT

Larvae of the invasive mosquito Culex quinquefasciatus Say are morphologically similar to those of the native Culex pervigilans Bergroth, yet distinguishing these species can be hampered by morphological variations in Cx. quinquefasciatus. We present detail about the extent of these variations in an urban population of Cx. quinquefasciatus in Auckland. To aid in identification of this exotic species, we provide images of key diagnostic characters and some observed exceptions to these. Details regarding variation in diagnostic characters for < 3rd instar and 3rd/4th instar larvae are given. Of the nine characters used for identification, three were highly consistent (dorsal papillae, mantle plate, pecten teeth); each observed in > 90% of larvae, although these characters were not always visible. Other characters were less reliable, for instance, the expected position of seta 1a-S in relation to the pecten teeth was observed in < 10% of larvae. Further exploration of regional morphological variation in both Cx. quinquefasciatus and Cx. pervigilans is recommended, ideally with associated molecular characterisation.     

Mosquitoes of the Jaú National Park and their potential importance in Brazilian Amazonia

Abstract. An entomological inventory was conducted between 1993 and 1996 to obtain information on the diversity of mosquitoes (Diptera: Culicidae) in the Jaú National Park, State of Amazonas, Brazil. A total of 10,159 adult (91%) and immature mosquito specimens, representing 130 taxa (species + morphospecies) in 16 genera, was collected. A species list for the family Culicidae is presented, including 30 new records for the State of Amazonas. The collecting localities were restricted to the alluvial subregion of the Open Tropical Forest found in the park. Most of the specimens (71%) were collected in forest habitats and the rest in areas of second growth and peridomicile. The majority of immature specimens (46%) were collected in bodies of water along the edges of rivers, flooded forests, lakes and streams. Among the various collection methods used, the Centre for Disease Control (CDC) and Shannon traps together were responsible for capturing 60% of the adults. More than 90% of the material collected belongs to the genera Culex (65%), Psorophora (19%), Wyeomyia (4%), and Anopheles (3%), which together represent 70% of the identified taxa. The genus Culex presented the largest number of species (45). The species Culex (Melanoconion) vaxus, Cx. (Mel.) pedroi, Psorophora amazonica, Cx. (Mel.) portesi and Cx. (Mel.) theobaldi together (< 4% of the recorded species) represent more than 63% of the material collected and identified to the species level. The most abundant species was Cx. (Mel.) vaxus, representing 17% of the material identified to species. The possible epidemiological and ecological implications of the species hereby recorded in the Jaú National Park are presented and discussed.     

云南南部新建村庄与现有村庄中三带喙库蚊与伪杂鳞库蚊种群动态的比较

对云南省南部新建移民东方红村和现有的龙塘村中三带喙库蚊与伪杂鳞库蚊种群动态情况进行了比较。结果表明:2个村庄中这2种媒介蚊虫的种群动态趋势不同;东方红村房屋内,除6月三带喙库蚊种群密度比伪杂鳞库蚊明显要高以外,其它月份二者种群密度差异不明显(P>0.05);整个诱集期间,伪杂鳞库蚊的种群密度(D)和种群优势度(d)在东方红村房屋(D=10只.灯-1,d=0.04)与牛舍(D=52只.灯-1,d=0.06)中都比龙塘村房屋(D=33只.灯-1,d=0.10)与牛舍(D=99只.灯-1,d=0.09)中的低。表明云南南部地区新移民村建设中,乙脑媒介种群随着环境开发产生变化。     

Nectar feeding by mosquitoes in Sweden, with special reference to Culex pipiens and Cx torrentium

1. Nectar feeding by mosquitoes collected from tansy (Tanacetum vulgare) flowers was studied in July and August 1983-85 at two sites in central and south-western Sweden. Prior to fructose analysis, gonotrophic state and parity was determined. 2. A total of 1010 mosquitoes (70% males) of eighteen species was collected. Fructose was detected in 75% of the males and 78% of the females. Most (86%) mosquitoes collected belonged to Culex pipiens and Cx torrentium. 3. Among 219 Cx pipiens females, for most of which the abdomen appeared gravid (52%) or empty (42%), 81% were fructose-positive. Females of other species were also proved to have fed on plant sugars in both early and later stages of the gonotrophic cycle. 4. In Cx pipiens and males of Cx torrentium peak nectar feeding occurred between 22.00 and 04.00 hours. Data for males of both species suggested a V-shaped pattern of nectar feeding activity during the night. 5. At about 16 degrees C nearly all mosquitoes became fructose-negative in the anthrone test within 20 h after collection from tansy flowers. 6. At least 44% of Cx pipiens females that were attracted to a dove-baited trap had fed on plant sugars shortly before.     

Behavioural response of the house mosquitoes Culex quinquefasciatus and Culex pipiens molestus to avian odours and its reliance on carbon dioxide

How Culex (Diptera: Culicidae) mosquitoes select and discriminate between potential avian hosts is critical for understanding the epidemiology of West Nile virus. Therefore, the present authors studied the behavioural responses of Culex quinquefasciatus (Say) and Culex pipiens molestus (Forsskål) to headspace volatiles of three avian species [chicken and pigeon (sexes analysed separately), and magpie], presented either alone or in combination with 600 p.p.m. carbon dioxide (CO₂). The attraction of Cx. quinquefasciatus to the headspace volatiles of both sexes of chicken, and of female pigeon, in combination with CO₂ was significantly higher than that achieved by the CO₂ and solvent control. Although Cx. p. molestus was attracted to headspace volatiles of chickens and magpies, it was repelled by those of female pigeons when combined with CO₂. An increased effect between the avian volatiles and CO₂ was observed for Cx. quinquefasciatus, whereas the addition of CO₂ had no effect on the attraction of Cx. p. molestus females. The results of this study demonstrate that Cx. quinquefasciatus and Cx. p. molestus are attracted to the odour of potential avian hosts. Future studies aimed at identifying the bioactive volatile compounds in the headspace of chickens may contribute to the potential development of effective surveillance and control tools against Culex species.     

The salivary gland chromosomes of Culex (Culex) vishnui (Culicidae: Diptera)

Culex vishnui is a member of the vishnui group comprising six closely resembling species. The salivary gland chromosomes are longer than those found in Anopheles, more fragile and spread with difficulty. The banding pattern is compared with that of Culex pipiens pipiens and Culex pipiens fatigans. Homologies are closest between vishnui and fatigans. The incidence of asynapsis between the homologues is very high.     

The effect of predatory fish exudates on the ovipostional behaviour of three mosquito species: Culex quinquefasciatus, Aedes aegypti and Culex tarsalis

Abstract.  Three mosquito species, Culex tarsalis Coquillett, Culex quinquefasciatus Say and Aedes aegypti L. (Diptera: Culicidae), were examined in laboratory binary choice experiments to investigate whether fish exudates from the mosquitofish, Gambusia affinis (Baird & Girard) (Cyprinodontiformes: Poecilliidae), deter oviposition and whether the responses of these mosquito species to fish exudates in oviposition sites are consistent with the risk of predation from fish experienced by each species in their respective natural breeding habitats. Culex tarsalis was deterred significantly from egg laying by the presence of fish exudates in oviposition cups, consistent with high levels of predation by fish in natural breeding sites. Egg laying by Cx quinquefasciatus was slightly reduced in water with fish exudates, but was not consistently deterred by water conditioned by mosquitofish, consistent with the species' relatively low risk of fish predation in natural habitats. Oviposition by container-breeding Ae. aegypti was not deterred by the presence of fish exudates in oviposition cups, consistent with a low risk of predation by fish in natural habitats.     

Septic tank mosquitoes: competition between species in central Nigeria

Exit traps, placed over the air vents of septic tanks, were used to examine species diversity and relative abundance of mosquitoes breeding in ammonia-rich waters of septic tanks. Of the six species found, Culex decens Theobald and Culex cinereus Theobald appeared to be competing successfully with Culex quinquefasciatus Say during the wet season but not during the long dry season. The seasonal timing of their displacement by Cx quinquefasciatus was variable and did not correlate well with climatic factors. The three other species present, generally during the wet season and early dry season, were Culex tigripes G. & C., Culex horridus Edwards and Aedes aegypt (L.). Experimental bucket ovitraps were used to assess preference towards covered (dark) septic tank water in comparison with sunlit septic tank water, covered and sunlit compost water. These were colonized by Cx quinquefasciatus, Cx decens, Ae. aegypti and Ae. vittatus Bigot. The covered septic tank water was more abundantly colonized by Cx quinquefasciatus and marginally so by the two Aedes species. Cx decens appeared to colonize the exposed compost water more readily in the dry season, but changed to the covered septic tank water in the wet season. The discussion centres around competition between these mosquitos species and concludes that it would be useful to know what environmental factors, or what aspects of competition, lead to severe natural reductions in the abundance of the major pest species Cx quinquefasciatus.     

Attraction of Culex pipiens to uropygial gland secretions does not explain feeding preference for American robins

Culex pipiens, the endemic mosquito vector of West Nile virus in eastern North America, is responsible for maintenance of the virus in avian reservoir hosts, the most important of which appears to be the American robin. One reason for the greater involvement of robins is believed to be the feeding preference of Cx. pipiens, however, the basis of this preference is not understood. We tested the hypothesis that the species‐specific chemical profile of avian uropygial gland secretions are used by Cx. pipiens as cues to locate birds and, therefore, may contribute to the observed feeding preferences. We used gas chromatography‐mass spectrometry to identify the semi‐volatile components of the uropygial gland secretions of American robins and two other common reservoir host species, the house sparrow and European starling. We found that the chemical composition of the robin secretions was different from those of the sparrows and starlings. Through behavioral choice trials conducted in a dual‐port olfactometer, we also found that Cx. pipiens did not prefer the secretions of robins over the other two species. Surprisingly, however, we found that Cx pipiens were more often attracted to live starlings over robins and to the secretions of starlings over those of robins.     

Culex saltanensis Dyar, 1928--natural vector of Plasmodium juxtanucleare in Rio de Janeiro, Brazil.

Searching for the natural vector of Plasmodium juxtanucleare in an enzootic locality: Granjas Calábria (33% of the chickens infected), Jacarepaguá, in Rio de Janeiro, Brazil, 13 comparative captures of mosquitoes were carried out, simultaneously on man (out-doors) and on chicken (in a poultry-yard), between 6 and 9 p.m., from September 1988 to March 1989. Culex saltanensis was the most frequent species in captures on chicken, accounting for 41.7% of the mosquitoes collected on this bait, showing to be highly ornithophilic (90% captured on chicken versus 10% on man). Seven specimens of Cx. saltanensis were found naturally infected in Granjas Calábria: five with mature pedunculate oocysts and two with sporozoites (one in the haemocoele and one in the salivary glands). These sporozoites produced an infection by P. juxtanucleare in a chick, which had parasitemia on day 41 after inoculation. One Cx. coronator was found with mature pedunculate oocysts. Culex saltanensis was regarded as primary vector of P. juxtanucleare in Rio de Janeiro for being highly ornithophilic and in enough density to maintain the transmission, having been found with infective sporozoites in its salivary glands, and being susceptible to the parasite and able to transmit experimentally it by the bite.     

Trapping biases of Culex torrentium and Culex pipiens revealed by comparison of captures in CDC traps,ovitraps, and gravid traps

We evaluate three trapping methods for their effectiveness at capturing Culex pipiens and Culex torrentium, both enzootic vectors of bird‐associated viruses in Europe. The comparisons, performed in two regions in Sweden, were among CDC traps baited with carbon dioxide, gravid traps, and ovitraps baited with hay infusion. The proportions of the two Culex species in a catch differed between trap types, with CDC traps catching a lower proportion of Cx. torrentium than both gravid traps and ovitraps. Between gravid traps and ovitraps, there was no difference in the proportions of the two species. The results indicate that Cx. torrentium may go undetected or underestimated compared to Cx. pipiens when using carbon dioxide baited CDC traps. The new insight of trap bias presented here adds an important dimension to consider when investigating these vectors of bird‐associated viruses in the field.