Invertebrate host–parasite associations are one of the keystones in order to understand vector-borne diseases. The study of these specific interactions provides information not only about how the vector is affected by the parasite at the gene-expression level, but might also reveal mosquito strategies for blocking the transmission of the parasites. A very well-known vector for human malaria is Anopheles gambiae. This mosquito species has been the main focus for genomics studies determining essential key genes and pathways over the course of a malaria infection. However, to-date there is an important knowledge gap concerning other non-mammophilic mosquito species, for example some species from the Culex genera which may transmit avian malaria but also zoonotic pathogens such as West Nile virus. From an evolutionary perspective, these 2 mosquito genera diverged 170 million years ago, hence allowing studies in both species determining evolutionary conserved genes essential during malaria infections, which in turn might help to find key genes for blocking malaria cycle inside the mosquito. Here, we extensively review the current knowledge on key genes and pathways expressed in Anopheles over the course of malaria infections and highlight the importance of conducting genomic investigations for detecting pathways in Culex mosquitoes linked to infection of avian malaria. By pooling this information, we underline the need to increase genomic studies in mosquito–parasite associations, such as the one in Culex–Plasmodium, that can provide a better understanding of the infection dynamics in wildlife and reduce the negative impact on ecosystems.

While the desire for systemic repellents is high, ineffective repellents put one at risk of insect-vectored pathogens. Vitamin B1, or thiamine, has been touted as a systemic insect repellent since 1943, and denounced as an ineffective placebo for just as long. This paper presents a scoping review of 104 relevant case reports, research studies, and review articles to trace the evolution of this idea and identify an evidence-based, scientific consensus. Reports of thiamine's systemic repellency are primarily anecdotal and based on uncontrolled trials and/or used bite symptoms as a proxy for reduced biting. Controlled experiments on insect landing and feeding found no evidence of repellency. Of the 49 relevant review papers, 16 insect bite prevention guidelines, and 4 government documents, none after the 1990s claimed thiamine is a repellent. The findings of this review are that thiamine cannot repel arthropods in any dosage or route of administration. Due to limited available evidence, the possibility that thiamine reduces the subjective symptoms of insect bites cannot currently be ruled out. Unfortunately, many medical professionals and travelers today still believe thiamine may be effective despite the evidence stating otherwise. Continued promotion of debunked repellents on the commercial market poses a serious risk in countries with the endemic, mosquito-vectored disease.

From 1971 to 2012, in New York State, years with human Eastern equine encephalitis (EEE) were more strongly associated with the presence of Aedes canadensis, Coquillettidia perturbans and Culiseta melanura mosquitoes infected with the EEE virus (Fisher's exact test, one-sided P = 0.005, 0.03, 0.03) than with Culiseta morsitans, Aedes vexans, Culex pipiens-restuans, Anopheles quadrimaculatus or Anopheles punctipennis (P = 0.05, 0.40, 0.33, 1.00, 1.00). The estimated relative risk of a case in a year in which the virus was detected vs. not detected was 14.67 for Ae. canadensis, 6.38 for Cq. perturbans and 5.50 for Cs. morsitans. In all 5 years with a case, Cs. melanura with the virus was detected. In no year was there a case in the absence of Cs. melanura with the virus. There were 18 years with no case in the presence of Cs. melanura with the virus. Such observations may identify the time of increased risk, and when the methods may be used to prevent or reduce exposure to vector mosquito species in this geographic region.

Factors such as the particular combination of parasite–mosquito species, their co-evolutionary history and the host's parasite load greatly affect parasite transmission. However, the importance of these factors in the epidemiology of mosquito-borne parasites, such as avian malaria parasites, is largely unknown. Here, we assessed the competence of two mosquito species [Culex pipiens and Aedes (Ochlerotatus) caspius], for the transmission of four avian Plasmodium lineages (Plasmodium relictum SGS1 and GRW11 and Plasmodium cathemerium-related lineages COLL1 and PADOM01) naturally infecting wild house sparrows. We assessed the effects of parasite identity and parasite load on Plasmodium transmission risk through its effects on the transmission rate and mosquito survival. We found that Cx. pipiens was able to transmit the four Plasmodium lineages, while Ae. caspius was unable to transmit any of them. However, Cx. pipiens mosquitoes fed on birds infected by P. relictum showed a lower survival and transmission rate than those fed on birds infected by parasites related to P. cathemerium. Non-significant associations were found with the host–parasite load. Our results confirm the existence of inter- and intra-specific differences in the ability of Plasmodium lineages to develop in mosquito species and their effects on the survival of mosquitoes that result in important differences in the transmission risk of the different avian malaria parasite lineages studied.

Mosquitoes are haematophagous vectors for hundreds of pathogenic viruses that are aetiological agents of human diseases. In nature, mosquito-borne viruses maintain a lifecycle between mosquitoes and vertebrate animals. Viruses are acquired by a naive mosquito from an infected host by blood meals and then propagate extensively in the mosquito's tissues. This mosquito then becomes a virus reservoir and is competent to transmit the viruses to a naive vertebrate host through the next blood meal. To survive in and efficiently cycle between two distinct host environments, mosquito-borne viruses have evolved delicate and smart strategies to comprehensively exploit host and vector factors. Here, we provide an update on recent studies of the mechanisms of virus survival in, acquisition and transmission by mosquitoes.

Australian mosquito species significantly impact human health through nuisance biting and the transmission of endemic and exotic pathogens. Surveillance programmes designed to provide an early warning of mosquito-borne disease risk require reliable identification of mosquitoes. This study aimed to investigate the viability of Matrix-Assisted Laser Desorption/Ionization–Time-of-Flight Mass Spectrometry (MALDI-TOF MS) as a rapid and inexpensive approach to the identification of Australian mosquitoes and was validated using a three-step taxonomic approach. A total of 300 mosquitoes representing 21 species were collected from south-eastern New South Wales and morphologically identified. The legs from the mosquitoes were removed and subjected to MALDI-TOF MS analysis. Fifty-eight mosquitoes were sequenced at the cytochrome c oxidase subunit I (cox1) gene region and genetic relationships were analysed. We create the first MALDI-TOF MS spectra database of Australian mosquito species including 19 species. We clearly demonstrate the accuracy of MALDI-TOF MS for identification of Australian mosquitoes. It is especially useful for assessing gaps in the effectiveness of DNA barcoding by differentiating closely related taxa. Indeed, cox1 DNA barcoding was not able to differentiate members of the Culex pipiens group, Cx. quinquefasciatus and Cx. pipiens molestus, but these specimens were correctly identified using MALDI-TOF MS.

Among the invasive mosquitoes registered all over the world, Aedes species are particularly frequent and important. As several of them are potential vectors of disease, they present significant health concerns for 21st century Europe. Five species have established in mainland Europe, with two (Aedes albopictus and Aedes japonicus) becoming widespread and two (Ae. albopictus and Aedes aegypti) implicated in disease transmission to humans in Europe. The routes of importation and spread are often enigmatic, the ability to adapt to local environments and climates are rapid, and the biting nuisance and vector potential are both an ecomonic and public health concern. Europeans are used to cases of dengue and chikungunya in travellers returning from the tropics, but the threat to health and tourism in mainland Europe is substantive. Coupled to that are the emerging issues in the European overseas territorities and this paper is the first to consider the impacts in the remoter outposts of Europe. If entomologists and public health authorities are to address the spread of these mosquitoes and mitigate their health risks they must first be prepared to share information to better understand their biology and ecology, and share data on their distribution and control successes. This paper focusses in greater detail on the entomological and ecological aspects of these mosquitoes to assist with the risk assessment process, bringing together a large amount of information gathered through the ECDC VBORNET project.

Insect population dynamics depend strongly on environmental factors. For floodwater mosquitoes, meteorological conditions are crucial in the rhythm of mosquito abundances. Indeed, rainfall triggers the egg hatching after flooding breeding sites, and temperature controls the duration of the aquatic immature development up to adult emergence.

According to this, we have developed a simple mechanistic and tractable model that describes the population dynamics of floodwater mosquitoes as a function only of the most accessible meteorological variables, rainfall and temperature. The model involves three parameters: development duration tdev of the immature aquatic stages, the adult emergence rate function f(t) (characterized by the emergence time scale τ and shaping the profile of adult population abundance), and the depletion rate, α, of adult disappearance.

The developed model was subsequently applied to fit experimental field data of the dynamics of Aedes caspius (Pallas), the main pest mosquito in southern France. First, it was found that the emergence rate function of adult mosquitoes very well reproduce experimental data of the dynamics of immature development for all sampled temperatures. The estimated values of tdev and τ both exhibit Arrhenius behaviour as a function of temperature. Second, using the meteorological records of rainfall and temperature as inputs, the model correctly fit data from a two-site CO2 trapping survey conducted in 2004 and 2005. The estimated depletion rates (summation of the mortality and the emigration rates) were found to be a concave quadratic function of temperature with a maximum of 0.5 per days at about 22°C.

Thirty-four spedes of mosquitoes (Diptera: Culicidae) were collected at a series of human-bait catches designed to study evening crepuscular biting activity using the Nielsen crep unit, at a site at Peradeniya, Sri Lanka. Crepuscular biting profiles of seven species (Armigeres subalbatus, Aedes albopictus, Culex pseudovishnui, Cx. gelidus, Cx. fuscocephala, Cx. tritaeniorhynchus and Mansonia uniformis) were obtained. Modal periods for the onset of biting were analysed in six species: Cx. pseudovishnui, Cx. fuscocephala, Cx. gelidus, Cx. tritaeniorhynchus, Cx. quinquefasciatus and Ma. uniformis. Successions in the biting peaks and in modal onset of biting were seen in these species. Analysis of biting in Ar. subalbatus and Cx. pseudovishnui under high (clear sky) and low (cloud-overcast sky) light intensities showed earlier biting in both species under the latter conditions. The value of the Nielsen crep unit in making latitudinal comparisons is illustrated in Ma. uniformis, for which parallel data are available from Africa.

Two clones of the human malaria parasite Plasmodium falciparum, denoted 3D7 and HB3, were grown in vitro under conditions permitting the development of gametocytes. The two clones differ in their allelic forms of two antigen genes MSP1 and MSP2. The alleles can be distinguished as size differences of polymerase chain reaction (PCR) amplified fragments of repetitive regions of each gene. Mosquitoes (Anopheles stephensi) were fed on a mixture of these gametocytes. A total of 128 oocysts was isolated from the midguts of infected mosquitoes from 9 crossing experiments between the clones. DNA extracted from these oocysts was amplified by PCR. Oocysts which contained both alleles of each gene (MSP1 and MSP2) had developed from heterozygotes produced by cross-fertilization events between 3D7 and HB3 gametes. The remaining oocysts contained single alleles of each gene, in parent clone combinations, and these had developed from homozygotes formed by self-fertilizations. The results suggest that gametes in the original mixture fed to mosquitoes had undergone random mating.

On the basis of known DNA sequences of Dirofilaria repens and D. immitis we designed specific primers for the amplification by Polymerase Chain Reaction (PCR) of the DNA from the two species. The PCR-based identification was found to be unambiguous and allowed specific diagnosis of microfilariae in blood samples, of developing larvae in the mosquito vector and of immature adults in bioptic material, overcoming the serious constraints of the morphological separation of these filarial parasites at the pre-adult stages. The technique was found to be very sensitive and applicable to samples stored either dry or in various preservation media, with the exception of formalin. The reliable identification of D. repens and D. immitis from bioptic material is expected to greatly enhance the chances of detecting human infections and to further clarify the role of the two parasites as pathogens of man. The possibility of routine identification of developing larvae in the vector will substantially improve the perspectives for epidemiological investigations, particularly in Southern European regions, such as Italy, where the two nematode species are largely sympatric.

Programmes involving mosquito research and surveillance are normally focused on the study of aquatic larval stages, but sampling methods are varied and not systematized, which hinders the comparative analysis of ecological data. A standardized method for assessing the richness and abundance of mosquito larval populations of value for the analysis of mosquito diversity is presented. Based on the study of all the aquatic sites in a one hectare sample area with a proportional number of dips according to the size of the aquatic habitat, comparative data can be obtained on species richness and relative abundance of species found between pairs of sites or in the same area at different times. This technique provides information on the total mosquito fauna at each site, helps recognize species of medical importance and estimates the abundance of each species; parameters that are not estimated by the current entomological indexes used in surveillance programmes. The quality of the inventory is obtained by estimation of the efficiency effort. Procedures for calculating alpha, beta and gamma diversity are presented. The technique was validated in a natural and an urban zone at La Mancha, Veracruz, Mexico, over two years of sampling made during different climatic seasons.

The performance of Nzi traps for tabanids (Tabanus similis Macquart, T. quinquevittatus Wiedemann, Chrysops aberrans Philip, C. univittatus Macquart, C. cincticornis Walker, Hybomitra lasiophthalma (Macquart)), stable flies (Stomoxys calcitrans Linnaeus) (Diptera: Muscidae) and mosquitoes (Aedes) (Diptera: Culicidae) was investigated at various sites in Canada (Ontario, Alberta) and USA (Iowa, Florida, Louisiana). Traps made from selected fabrics, insect nettings and hand-dyed blue cotton were compared to the African design to provide practical recommendations for temperate environments. Comparisons of substituted materials showed that trap performance was optimal only when traps were made from appropriate fabrics in the colours produced by either copper phthalocyanine (phthalogen blue), or its sulphonated forms (turquoise). Fabrics dyed with other blue chromophores were not as effective (anthraquinone, disazo, formazan, indanthrone, triphenodioxazine). An appropriate texture as well as an appropriate colour was critical for optimal performance. Smooth, shiny synthetic fabrics (polyester, nylon) and polyester blends reduced catches. Low catches occurred even for nominal phthalogen blue, but slightly-shiny, polyester fabrics in widespread use for tsetse. The most suitable retail fabric in place of phthalogen blue cotton was Sunbrella Pacific Blue acrylic awning/marine fabric. It was both attractive and durable, and had a matching colour-fast black. Nzi traps caught grossly similar numbers of biting flies as canopy, Vavoua, and Alsynite cylinder traps, but with differences in relative performance among species or locations.

We have examined metabolic rate, lipid and carbohydrate of female Aedes aegypti during 10 days following a malaria-infected bloodmeal. In parallel, we determined bloodmeal size, portions retained and diuresed, and subsequent fecundity. We found that mosquitoes obtained identical masses of blood when feeding on an infected or control host. However, infected mosquitoes lost more mass during diuresis and retained a smaller mass. Infection led to a significant reduction in fecundity, the extent of which could not be explained by the difference in post-diuresis bloodmeal mass alone. We found no differences in lipid or carbohydrate content between infected and control mosquitoes during the 10 days post-infection, although infected mosquitoes had a lower body mass than controls. Metabolic rates were not different between groups, except during blood digestion, where the metabolic rate was lower in infected mosquitoes. These results suggest that infection by malaria does not lead to an increase in metabolic rate during the phases of midgut invasion and sporogony. However, infection does have a measurable effect on fecundity and subsequent body mass of the infected females.

Various morphological characters of larvae of Aedes (Stg.) simpsoni complex collected from different localities in Uganda were studied in an attempt to characterise the species in the Ae. simpsoni complex. Morphometrics of larval characters revealed wide ranges of variation most of which separated or grouped populations. Multivariate discriminant analysis and principal component analysis discriminated between different populations. In some populations the males were fairly distinct from the females, while for some characters the populations overlapped considerably. The percent variability of each character alone is low and does not separate the species of the complex. However several characters correlated. Length of antennae (LANT), length of antennae up to Seta 1–4 (LANT4), length of head (LHEAD), and length of siphon up to position of Seta 1-S (LSIPHSET1S) showed fairly significant variability which could be used in conjunction with characters of pupae and adults to separate populations. In the wild we find anthropophilic and non-anthropophilic populations of the mosquitoes, especially in Colocasia plants, bananas (Musa sp.), Dracaenia and Ananas sp. The multivariate analysis carried out could not separate the populations into biting behaviour groups nor breeding habitat groups.

Studies of invertebrate–parasite interactions frequently report that infection reduces host fecundity. The extent of the reduction is likely to be determined by a wide range of host and parasite factors. We conducted a laboratory experiment to evaluate the role of parasite genetics and infection genetic diversity on the fecundity of mosquitoes carrying malaria parasites. The malaria vector Anopheles stephensi was infected with either of 2 different genotypes of the rodent malaria parasite Plasmodium chabaudi, or by a mixture of both. Mixed genotype infections reduced mosquito fecundity by 20%, significantly more than either of the 2 single genotype infections. Mixed genotype infections were associated with high gametocyte densities and anaemia in mice, both of which were correlated with reduced bloodmeal size in mosquitoes. Bloodmeal size was the most important predictor of mosquito fecundity; the presence and number of parasites had no direct effect. Parasite density influenced the propensity of mosquitoes to feed on infected mice, with a higher percentage of mosquitoes taking a meal as asexual parasite and gametocyte density increased. Thus mosquitoes may preferentially feed on hosts who will most impair their fecundity.

A simple method for storing mosquito bloodmeal samples, which permits extraction and detection of human DNA after polymerase chain reaction (PCR) amplification of target DNA sequences, was tested. Abdomens of bloodfed field-collected Anopheles gambiae s.l. and An. funestus mosquitoes (Diptera: Culicidae) were directly expressed onto filter paper, air-dried and stored at room temperature. DNA was extracted and amplified at human hypervariable loci TC11, VWA and D1S80. The amplified products were separated using polyacrylamide gel electrophoresis, visualised by silver staining, and results compared with those from mosquitoes that had been preserved in liquid nitrogen. DNA from blooded abdomens stored on dried filter papers could be amplified with greater than 95 % success for any locus, storage temperature, mosquito species or storage duration. Collection and drying of mosquito bloodmeals directly onto filter paper appears to be a more convenient method for sample transportation and storage than the conventional method involving cryopreservation.