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de novo transcriptome sequencing and analysis of coccinella septempunctata l. in non-diapause, diapause and diapause-terminated states to identify diapause-associated genes.the most common ladybird beetle, coccinella septempunctata l., is an excellent predator of crop pests such as aphids and white flies, and it shows a wide range of adaptability, a large appetite and a high reproductive ability. diapause research plays an important role in the artificial propagation and shelf-life extension of insect products. although this lady beetle's regulatory, physiological and biochemical characteristics in the diapause period are well understood, the molecular mechanism of ...201526689283
host preferences in host-seeking and blood-fed mosquitoes in switzerland.the avian zoonotic agent for west nile virus (wnv) can cause neuroinvasive disease in horses and humans and is expanding its range in europe. analyses of the risk for transmission to these hosts in non-endemic areas are necessary. host preferences of mosquitoes (diptera: culicidae), the main vectors of wnv, were determined in switzerland using animal-baited trap (horse, chickens) experiments at a natural and a periurban site. this was undertaken on four occasions during may-september 2014. in ad ...201626685926
estimation of allele-specific ace-1 duplication in insecticide-resistant anopheles mosquitoes from west africa.identification of variation in ace-1 copy number and g119s mutation genotype from samples of anopheles gambiae and anopheles coluzzii across west africa are important diagnostics of carbamate and organophosphate resistance at population and individual levels. the most widespread and economical method, pcr-rflp, suffers from an inability to discriminate true heterozygotes from heterozygotes with duplication.201526682913
effects of a red marker dye on aedes and culex larvae: are there implications for operational mosquito control?marker dyes are often mixed with liquid insecticide formulations prior to field applications to accurately determine the characteristics and penetration of droplets into targeted habitats. we have been using fd&c red 40 granular dm food dye at the rate of 20 g/liter in liquid solutions of bacillus thuringiensis israelensis (bti) for area-wide larvicide applications against the asian tiger mosquito, aedes albopictus. the bti and dye mix ratio has been recommended by pesticide manufacturers for te ...201526675462
an updated checklist of the mosquitoes of oklahoma including new state records and west nile virus vectors, 2003-06.the mosquito fauna of oklahoma has not been evaluated since 1965 and no report has been published concerning species associated with urban areas in the state. mosquito collections were conducted as part of the west nile virus (wnv) surveillance program between april and november from 2003 to 2006, using standard collection methods. a total of 74,756 adults were collected in 26 urban centers in 16 counties of oklahoma. altogether, 40 species were recorded during this study period, bringing the to ...201526675455
molecular detection of six (endo-) symbiotic bacteria in belgian mosquitoes: first step towards the selection of appropriate paratransgenesis candidates.actually, the use of symbiotic bacteria is one of alternative solution to avoid vector resistance to pesticides. in belgium, among 31 identified mosquito species, 10 were considered as potential vectors. given to introduction risks of arbovirosis, the purpose of this study was to investigate the presence of symbiosis bacteria in potential mosquito vectors. eleven species caught from 12 sites in belgium were used: culex pipiens s.l., culex torrentium, culex hortensis, anopheles claviger, anophele ...201626670313
chemical composition and larvicidal activity of greek myrtle essential oils against culex pipiens biotype molestus.fresh leaves of myrtus communis collected from different localities in greece, were subjected to hydrodistillation and the oils obtained were analyzed by gc-fid and gc-ms. the analyses showed mainly quantitative differences, with the monoterpenes myrtenyl acetate, α-pinene, 1,8-cineole, and linalool, along with limonene, dominating the majority of the analyzed myrtle oils. the evaluation of the larvicidal activity of the samples against culex pipiens biotype molestus mosquito showed that all tes ...201526669120
sporogony and sporozoite rates of avian malaria parasites in wild culex pipiens pallens and c. inatomii in japan.malaria infection in mosquitoes is traditionally detected by microscopic examination for plasmodium oocysts and sporozoites. although pcr is now widely used, the presence of parasite dna in a mosquito does not prove that sporogony is achieved. thus, detection of sporozoites by microscopy is still required to definitively identify vector mosquitoes. the aim of this study was to confirm sporogony of avian plasmodium spp. in culex pipiens pallens and c. inatomii caught from the wild.201526666959
cardio-pulmonary parasitic nematodes affecting cats in europe: unraveling the past, depicting the present, and predicting the future.various cardio-pulmonary parasitic nematodes infecting cats have recently been fascinating and stimulating the attention of the academia, pharmaceutical companies, and veterinary practitioners. this is the case of the metastrongyloids: aelurostrongylus abstrusus and troglostrongylus brevior, the trichuroid: capillaria aerophila (syn. eucoleus aerophilus), and the filarioid: dirofilaria immitis. apparently, these parasites have been emerging in several european countries, thus, gaining an importa ...201426664917
direct pcr of indigenous and invasive mosquito species: a time- and cost-effective technique of mosquito barcoding.millions of people die each year as a result of pathogens transmitted by mosquitoes. however, the morphological identification of mosquito species can be difficult even for experts. the identification of morphologically indistinguishable species, such as members of the anopheles maculipennis complex (diptera: culicidae), and possible hybrids, such as culex pipiens pipiens/culex pipiens molestus (diptera: culicidae), presents a major problem. in addition, the detection and discrimination of newly ...201526663040
potential applications of insect symbionts in biotechnology.symbiotic interactions between insects and microorganisms are widespread in nature and are often the source of ecological innovations. in addition to supplementing their host with essential nutrients, microbial symbionts can produce enzymes that help degrade their food source as well as small molecules that defend against pathogens, parasites, and predators. as such, the study of insect ecology and symbiosis represents an important source of chemical compounds and enzymes with potential biotechn ...201526659224
(e)-caryophyllene and α-humulene: aedes aegypti oviposition deterrents elucidated by gas chromatography-electrophysiological assay of commiphora leptophloeos leaf oil.aedes aegypti is responsible for the transmission of dengue, a disease that infects millions of people each year. although essential oils are well recognized as sources of compounds with repellent and larvicidal activities against the dengue mosquito, much less is known about their oviposition deterrent effects. commiphora leptophloeos, a tree native to south america, has important pharmacological properties, but the chemical profile and applicability of its essential oil in controlling the spre ...201526650757
molecular differentiation of the african yellow fever vector aedes bromeliae (diptera: culicidae) from its sympatric non-vector sister species, aedes lilii.yellow fever continues to be a problem in sub-saharan africa with repeated epidemics occurring. the mosquito aedes bromeliae is a major vector of yellow fever, but it cannot be readily differentiated from its non-vector zoophilic sister species ae. lilii using morphological characters. genetic differences have been reported between anthropophilic ae. bromeliae and zoophilic ae. lilii and between forest and domestic populations. however, due to the application of different molecular markers and n ...201526641858
larvicidal activity of nerium oleander against larvae west nile vector mosquito culex pipiens (diptera: culicidae).background. outbreaks of the west nile virus infection were reported in morocco in 1996, 2003, and 2010. culex pipiens was strongly suspected as the vector responsible for transmission. in the north center of morocco, this species has developed resistance to synthetic insecticides. there is an urgent need to find alternatives to the insecticides as natural biocides. objective. in this work, the insecticidal activity of the extract of the local plant nerium oleander, which has never been tested b ...201526640701
arbovirus infections.arbovirus (arthropod-borne virus) infections are increasingly important causes of neurologic disease in the united states through both endemic transmission and travel-associated infections. this article reviews the major arbovirus infections that can cause neurologic disease likely to be encountered in the united states.026633778
arbovirus-mosquito interactions: rnai pathway.arthropod-borne (arbo) viruses infect hematophagous arthropods (vectors) to maintain virus transmission between vertebrate hosts. the mosquito vector actively controls arbovirus infection to minimize its fitness costs. the rna interference (rnai) pathway is the major antiviral response vectors use to restrict arbovirus infections. we know this because depleting rnai gene products profoundly impacts arbovirus replication, the antiviral rnai pathway genes undergo positive, diversifying selection a ...201526629932
fauna and larval habitat characteristics of mosquitoes in neka county, northern iran.ecological studies on mosquitoes are very important in vector control programs. there are a few studies about the ecology of mosquitoes in northern iran. this study was carried out to detect fauna and larval habitat characteristics of mosquitoes.201526623437
dissolved oxygen levels affect the survival and developmental period of the mosquito culex pipiens. 201526611983
detection of persistent chikungunya virus rna but not infectious virus in experimental vertical transmission in aedes aegypti from malaysia.vertical transmission may contribute to the maintenance of arthropod-borne viruses, but its existence in chikungunya virus (chikv) is unclear. experimental vertical transmission of infectious clones of chikv in aedes aegypti mosquitoes from malaysia was investigated. eggs and adult progeny from the second gonotrophic cycles of infected parental mosquitoes were tested. using polymerase chain reaction (pcr), 56.3% of pooled eggs and 10% of adult progeny had detectable chikv rna, but no samples had ...201526598564
comparison of sampling techniques for rift valley fever virus potential vectors, aedes aegypti and culex pipiens complex, in ngorongoro district in northern tanzania.we investigated mosquito sampling techniques with two types of traps and attractants at different time for trapping potential vectors for rift valley fever virus. the study was conducted in six villages in ngorongoro district in tanzania from september to october 2012. a total of 1814 mosquitoes were collected, of which 738 were collected by cdc light traps and 1076 by mosquito magnet trapping technique. of the collected mosquitoes, 12.46% (n = 226) were aedes aegypti and 87.54% (n = 1588) were ...201326591704
genomic analysis of detoxification supergene families in the mosquito anopheles sinensis.anopheles sinensis is an important malaria vector in china and other southeast asian countries, and the emergence of insecticide resistance in this mosquito poses a serious threat to the efficacy of malaria control programs. the recently published an. sinensis genome and transcriptome provide an opportunity to understand the molecular mechanisms of insecticide resistance. analysis of the an. sinensis genome revealed 174 detoxification genes, including 93 cytochrome p450s (p450s), 31 glutathione- ...201526588704
deltamethrin resistance mechanisms in aedes aegypti populations from three french overseas territories worldwide.aedes aegypti is a cosmopolite mosquito, vector of arboviruses. the worldwide studies of its insecticide resistance have demonstrated a strong loss of susceptibility to pyrethroids, the major class of insecticide used for vector control. french overseas territories such as french guiana (south america), guadeloupe islands (lesser antilles) as well as new caledonia (pacific ocean), have encountered such resistance.201526588076
viral interference and persistence in mosquito-borne flaviviruses.mosquito-borne flaviviruses are important pathogens for humans, and the detection of two or more flaviviruses cocirculating in the same geographic area has often been reported. however, the epidemiological impact remains to be determined. mosquito-borne flaviviruses are primarily transmitted through aedes and culex mosquitoes; these viruses establish a life-long or persistent infection without apparent pathological effects. this establishment requires a balance between virus replication and the ...201526583158
behavioural response of female culex pipiens pallens to common host plant volatiles and synthetic blends.most mosquito species need to obtain sugar from host plants. little is known about the chemical cues that culex pipiens pallens use during their orientation to nectar host plants. in this study, we investigated the behavioural responses of female cx. pipiens pallens to common floral scent compounds and their blends.201526577584
yeasts associated with culex pipiens and culex theileri mosquito larvae and the effect of selected yeast strains on the ontogeny of culex pipiens.the success of mosquitoes in nature has been linked to their microbiota and bacteria in particular. yet, knowledge on their symbioses with yeasts is lacking. to explore possible associations, culturable yeasts were isolated from wild larvae of culex pipiens and culex theileri. these yeasts were classified using restriction fragment length polymorphism (rflp) analyses and identified by sequencing the d1/d2 region of the 26s rrna gene. representative strains of candida, cryptococcus, galactomyces, ...201626573833
dissecting vectorial capacity for mosquito-borne viruses.the inter-relationship between mosquitoes and the viruses they transmit is complex. while previously understood barriers to infection and transmission remain valid, additional factors have been uncovered that suggest an 'arms race' between mosquito and virus. these include the mosquito microbiota and interplay between mosquito and viral genetics. following an infectious blood meal, the mosquito mounts an immune and transcriptional response, leading to altered expression of multiple genes. these ...201526569343
potential for co-infection of a mosquito-specific flavivirus, nhumirim virus, to block west nile virus transmission in mosquitoes.nhumirim virus (nhuv) is an insect-specific virus that phylogenetically affiliates with dual-host mosquito-borne flaviviruses. previous in vitro co-infection experiments demonstrated prior or concurrent infection of aedes albopictus c6/36 mosquito cells with nhuv resulted in a 10,000-fold reduction in viral production of west nile virus (wnv). this interference between wnv and nhuv was observed herein in an additional ae. albopictus mosquito cell line, c7-10. a wnv 2k peptide (v9m) mutant capabl ...201526569286
the impact of wolbachia on virus infection in mosquitoes.mosquito-borne viruses such as dengue, west nile and chikungunya viruses cause significant morbidity and mortality in human populations. since current methods are not sufficient to control disease occurrence, novel methods to control transmission of arboviruses would be beneficial. recent studies have shown that virus infection and transmission in insects can be impeded by co-infection with the bacterium wolbachia pipientis. wolbachia is a maternally inherited endosymbiont that is commonly found ...201526556361
testing local adaptation in a natural great tit-malaria system: an experimental approach.finding out whether plasmodium spp. are coevolving with their vertebrate hosts is of both theoretical and applied interest and can influence our understanding of the effects and dynamics of malaria infection. in this study, we tested for local adaptation as a signature of coevolution between malaria blood parasites, plasmodium spp. and its host, the great tit, parus major. we conducted a reciprocal transplant experiment of birds in the field, where we exposed birds from two populations to plasmo ...201526555892
metallic copper spray--a new control technique to combat invasive container-inhabiting mosquitoes.the control of container-inhabiting mosquitoes is mainly based on environmental management with special emphasis on community participation e.g. source reduction by elimination or modification of water bodies. however, citizens are often not aware of the problems related to urban mosquito control or just ignore the advice provided during anti-mosquito control campaigns. in particular, cemeteries contain favourite breeding sites for container-inhabiting mosquitoes like ochlerotatus j. japonicus, ...201526553319
the ecological foundations of transmission potential and vector-borne disease in urban landscapes.urban transmission of arthropod-vectored disease has increased in recent decades. understanding and managing transmission potential in urban landscapes requires integration of sociological and ecological processes that regulate vector population dynamics, feeding behavior, and vector-pathogen interactions in these unique ecosystems. vectorial capacity is a key metric for generating predictive understanding about transmission potential in systems with obligate vector transmission. this review eva ...201526549921
insect biometrics: optoacoustic signal processing and its applications to remote monitoring of mcphail type traps.monitoring traps are important components of integrated pest management applied against important fruit fly pests, including bactrocera oleae (gmelin) and ceratitis capitata (widemann), diptera of the tephritidae family, which effect a crop-loss/per year calculated in billions of euros worldwide. pests can be controlled with ground pesticide sprays, the efficiency of which depends on knowing the time, location and extent of infestations as early as possible. trap inspection is currently carried ...201526544845
bacterial calpains and the evolution of the calpain (c2) family of peptidases.homologues of calpain, often thought to be an essential, cytoplasmic, calcium-dependent cysteine endopeptidase found exclusively in eukaryotes, have been found in bacterial proteomes. the homologues lack calcium-binding sites, have differing domain architectures, and can be secreted or membrane-associated. homologues are rare and occur in a minority of bacterial phyla and often in a minority of species in a genus. however, the differences in domain architecture argue against a recent, horizontal ...201526527411
comparison of repellency effect of mosquito repellents for deet, citronella, and fennel oil.to confirm that korean food and drug administration (kfda) guidelines are applicable to test the efficacy of mosquito repellents, these guidelines were used to test the efficacy and complete protection times (cpts) of three representative mosquito repellents: n,n-diethyl-3-methylbenzamide (deet), citronella, and fennel oil. the repellency of citronella oil decreased over time, from 97.9% at 0 h to 71.4% at 1 h and 57.7% at 2 h, as did the repellency of fennel oil, from 88.6% at 0 h to 61.2% at 1 ...201526527362
the evolutionary landscape of intergenic trans-splicing events in insects.to explore the landscape of intergenic trans-splicing events and characterize their functions and evolutionary dynamics, we conduct a mega-data study of a phylogeny containing eight species across five orders of class insecta, a model system spanning 400 million years of evolution. a total of 1,627 trans-splicing events involving 2,199 genes are identified, accounting for 1.58% of the total genes. homology analysis reveals that mod(mdg4)-like trans-splicing is the only conserved event that is co ...201526521696
genotypic and phenotypic analyses of a pseudomonas aeruginosa chronic bronchiectasis isolate reveal differences from cystic fibrosis and laboratory strains.pseudomonas aeruginosa is an environmentally ubiquitous gram-negative bacterium and important opportunistic human pathogen, causing severe chronic respiratory infections in patients with underlying conditions such as cystic fibrosis (cf) or bronchiectasis. in order to identify mechanisms responsible for adaptation during bronchiectasis infections, a bronchiectasis isolate, pahm4, was phenotypically and genotypically characterized.201526519161
evaluation of the pathogenicity of west nile virus (wnv) lineage 2 strains in a spf chicken model of infection: ns3-249pro mutation is neither sufficient nor necessary for conferring virulence.lineage 2 west nile virus (wnv) strains were reported for the first time in europe in 2004. despite an almost silent circulation around their entry point in hungary, an upsurge of pathogenicity occurred in 2010 as 262 people suffered from neuroinvasive disease in greece. this increase in virulence was imputed to the emergence of a his249pro mutation in the viral ns3 helicase, as previously evidenced in american crows experimentally infected with the prototype lineage 1 north-american wnv strain. ...201526518144
detection of oropouche virus segment s in patients and inculex quinquefasciatus in the state of mato grosso, brazil.this study aimed to investigate the circulation of orthobunyavirus species in the state of mato grosso (mt) brazil. during a dengue outbreak in 2011/2012, 529 serum samples were collected from patients with acute febrile illness with symptoms for up to five days and 387 pools of female culex quinquefasciatus captured in 2013 were subjected to nested-reverse transcription-polymerase chain reaction for segment s of the simbu serogroup followed by nucleotide sequencing and virus isolation in vero c ...026517653
exposure to west nile virus increases bacterial diversity and immune gene expression in culex pipiens.complex interactions between microbial residents of mosquitoes and arboviruses are likely to influence many aspects of vectorial capacity and could potentially have profound effects on patterns of arbovirus transmission. such interactions have not been well studied for west nile virus (wnv; flaviviridae, flavivirus) and culex spp. mosquitoes. we utilized next-generation sequencing of 16s ribosomal rna bacterial genes derived from culex pipiens linnaeus following wnv exposure and/or infection and ...201526516902
decreased flight activity in culex pipiens (diptera: culicidae) naturally infected with culex flavivirus.insect-specific flaviviruses (isfvs) commonly infect vectors of mosquito-borne arboviruses. to investigate whether infection with an isfv might affect mosquito flight behavior, we quantified flight behavior in culex pipiens l. naturally infected with culex flavivirus (cxfv). we observed a significant reduction in the scotophase (dark hours) flight activity of cxfv-positive mosquitoes relative to cxfv-negative mosquitoes, but only a marginal reduction in photophase (light hours) flight activity, ...201626512141
dna barcoding of neotropical sand flies (diptera, psychodidae, phlebotominae): species identification and discovery within brazil.dna barcoding has been an effective tool for species identification in several animal groups. here, we used dna barcoding to discriminate between 47 morphologically distinct species of brazilian sand flies. dna barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23-19.04%), indicating cryptic diversity. ...201526506007
dynamics of wolbachia pipientis gene expression across the drosophila melanogaster life cycle.symbiotic interactions between microbes and their multicellular hosts have manifold biological consequences. to better understand how bacteria maintain symbiotic associations with animal hosts, we analyzed genome-wide gene expression for the endosymbiotic α-proteobacteria wolbachia pipientis across the entire life cycle of drosophila melanogaster. we found that the majority of wolbachia genes are expressed stably across the d. melanogaster life cycle, but that 7.8% of wolbachia genes exhibit rob ...201526497146
modeling occurrence of urban mosquitos based on land use types and meteorological factors in korea.mosquitoes are a public health concern because they are vectors of pathogen, which cause human-related diseases. it is well known that the occurrence of mosquitoes is highly influenced by meteorological conditions (e.g., temperature and precipitation) and land use, but there are insufficient studies quantifying their impacts. therefore, three analytical methods were applied to determine the relationships between urban mosquito occurrence, land use type, and meteorological factors: cluster analys ...201526492260
mitochondrial genomes and comparative analyses of culex camposi, culex coronator, culex usquatus and culex usquatissimus (diptera:culicidae), members of the coronator group.the coronator group currently encompasses six morphologically similar species (culex camposi dyar, culex coronator dyar and knab, culex covagarciai forattini, culex usquatus dyar, culex usquatissimus dyar, and culex ousqua dyar). culex coronator has been incriminated as a potential vector of west nile virus (wnv), saint louis encephalitis virus (slev), and venezuelan equine encephalitis virus (veev). the complete mitochondrial genome of cx. coronator, cx. usquatus, cx.usquatissimus, and cx. camp ...201526489754
west nile virus surveillance in 2013 via mosquito screening in northern italy and the influence of weather on virus circulation.west nile virus (wnv) is a recently re-emerged health problem in europe. in italy, an increasing number of outbreaks of west nile disease, with occurrences of human cases, have been reported since 2008. this is particularly true in northern italy, where entomological surveillance systems have been implemented at a regional level. the aim of this study was to use, for the first time, all the entomological data collected in the five regions undergoing surveillance for wnv in northern italy to char ...201526488475
repellent effect of ocimum basilicum and glycyrrhiza glabra extracts against the mosquito vector, culex pipiens (diptera: culicidae).essential or volatile oils of plants have been variously reported to have many medicinal applications. methanol, acetone and petroleum ether extracts of ocimum basilicum and glycyrrhiza glabra were screened for their repellency effect against culex pipiens mosquito. the repellent action of the present plants extracts were varied depending on the solvent used and dose of extract. methanol extract of o. basilicum exhibited the lowest repellent activity as it recorded 77.4% at 6.7mg/cm2. the petrol ...201526485843
experimental and natural infections of goats with severe fever with thrombocytopenia syndrome virus: evidence for ticks as viral vector.severe fever with thrombocytopenia syndrome virus (sftsv), the causative agent for the fatal life-threatening infectious disease, severe fever with thrombocytopenia syndrome (sfts), was first identified in the central and eastern regions of china. although the viral rna was detected in free-living and parasitic ticks, the vector for sftsv remains unsettled.201526485390
identification of qtls conferring resistance to deltamethrin in culex pipiens pallens.culex pipiens pallens is the most abundant culex mosquito species in northern china and is an important vector of bancroftian filariasis and west nile virus. deltamethrin is an insecticide that is widely used for mosquito control, however resistance to this and other insecticides has become a major challenge in the control of vector-borne diseases that appear to be inherited quantitatively. furthermore, the genetic basis of insecticide resistance remains poorly understood. in this study, quantit ...201526484540
how do nutritional stress and la crosse virus infection interact? tests for effects on willingness to blood feed and fecundity in aedes albopictus (diptera: culicidae).evolutionary theory predicts that vector-borne pathogens should have low virulence for their vector because of selection against pathogens that harm the vector sufficiently to reduce transmission. environmental factors such as nutritional stress can alter vector-pathogen associations by making the vectors more susceptible to pathogens (condition-dependent competence) and vulnerable to the harm caused by pathogen replication (condition-dependent virulence). we tested the hypotheses of condition-d ...201526477049
age and prior blood feeding of anopheles gambiae influences their susceptibility and gene expression patterns to ivermectin-containing blood meals.ivermectin has been proposed as a novel malaria transmission control tool based on its insecticidal properties and unique route of acquisition through human blood. to maximize ivermectin's effect and identify potential resistance/tolerance mechanisms, it is important to understand its effect on mosquito physiology and potential to shift mosquito population age-structure. we therefore investigated ivermectin susceptibility and gene expression changes in several age groups of female anopheles gamb ...201526471037
effects of host sex, plant species, and putative host species on the prevalence of wolbachia in natural populations of bemisia tabaci (hemiptera: aleyrodidae): a modified nested pcr study.bemisia tabaci (gennadius) (hemiptera: aleyrodidae) is a globally distributed pest. one of the key endosymbionts in b. tabaci is wolbachia, an α-proteobacterium implicated in many important biological processes. previous studies indicated that the infection frequency of wolbachia in middle east-asia minor 1 (meam1) and mediterranean (med) varied greatly among populations in different areas. however, little is known about the factors that influence the prevalence of wolbachia in b. tabaci. in thi ...201526470122
parameters of mosquito-enhanced west nile virus infection.the arthropod-borne west nile virus (wnv) emerged in new york state in 1999 and quickly spread throughout the united states. transmission is maintained in an enzootic cycle in which infected mosquitoes transmit the virus to susceptible hosts during probing and feeding. arthropod-derived components within the viral inoculum are increasingly acknowledged to play a role in infection of vertebrate hosts. we previously showed that culex tarsalis mosquito saliva and salivary gland extract (sge) enhanc ...201526468544
molecular mechanisms of transcription activation by juvenile hormone: a critical role for bhlh-pas and nuclear receptor proteins.juvenile hormone (jh) is responsible for controlling many biological processes. in several insect species jh has been implicated as a key regulator of developmental timing, preventing the premature onset of metamorphosis during larval growth periods. however, the molecular basis of jh action is not well-understood. in this review, we highlight recent advances which demonstrate the importance of transcription factors from the bhlh-pas and nuclear receptor families in mediating the response to jh.201226467963
modification of insect and arachnid behaviours by vertically transmitted endosymbionts: infections as drivers of behavioural change and evolutionary novelty.vertically acquired, endosymbiotic bacteria such as those belonging to the rickettsiales and the mollicutes are known to influence the biology of their arthropod hosts in order to favour their own transmission. in this study we investigate the influence of such reproductive parasites on the behavior of their insects and arachnid hosts. we find that changes in host behavior that are associated with endosymbiont infections are not restricted to characteristics that are directly associated with rep ...201226467958
insect sex determination manipulated by their endosymbionts: incidences, mechanisms and implications.the sex-determining systems of arthropods are surprisingly diverse. some species have male or female heterogametic sex chromosomes while other species do not have sex chromosomes. most species are diploids but some species, including wasps, ants, thrips and mites, are haplodiploids (n in males; 2n in females). many of the sexual aberrations, such as sexual mosaics, sex-specific lethality and conversion of sexuality, can be explained by developmental defects including double fertilization of a bi ...201226467955
metabolic resistance in bed bugs.blood-feeding insects have evolved resistance to various insecticides (organochlorines, pyrethroids, carbamates, etc.) through gene mutations and increased metabolism. bed bugs (cimex lectularius) are hematophagous ectoparasites that are poised to become one of the major pests in households throughout the united states. currently, c. lectularius has attained a high global impact status due to its sudden and rampant resurgence. resistance to pesticides is one factor implicated in this phenomenon. ...201126467498
comparative study on the effectiveness of different mosquito traps in arbovirus surveillance with a focus on wnv detection.the selection of the ideal trap for arbovirus surveillance is an issue of primary importance to increase the sensitivity of virus detection and the cost-effectiveness of the entomological surveillance. during the summer 2011, the effectiveness of five types of mosquito traps (cdc gravid trap, co2(-)baited trap, bg-sentinel™ and two experimental prototypes) to attract females potentially infected with west nile virus were assessed. the study was carried out in three natural wetland sites located ...201626466982
new insights into control of arbovirus replication and spread by insect rna interference pathways.arthropod-borne (arbo) viruses are transmitted by vectors, such as mosquitoes, to susceptible vertebrates. recent research has shown that arbovirus replication and spread in mosquitoes is not passively tolerated but induces host responses to control these pathogens. small rna-mediated host responses are key players among these antiviral immune strategies. studies into one such small rna-mediated antiviral response, the exogenous rna interference (rnai) pathway, have generated a wealth of informa ...201226466541
reciprocal trophic interactions and transmission of blood parasites between mosquitoes and frogs.the relationship between mosquitoes and their amphibian hosts is a unique, reciprocal trophic interaction. instead of a one-way, predator-prey relationship, there is a cyclical dance of avoidance and attraction. this has prompted spatial and temporal synchrony between organisms, reflected in emergence time of mosquitoes in the spring and choice of habitat for oviposition. frog-feeding mosquitoes also possess different sensory apparatuses than do their mammal-feeding counterparts. the reciprocal ...201226466534
laboratory and simulated field bioassays to evaluate larvicidal activity of pinus densiflora hydrodistillate, its constituents and structurally related compounds against aedes albopictus, aedes aegypti and culex pipiens pallens in relation to their inhibitory effects on acetylcholinesterase activity.the toxicity of pinus densiflora (red pine) hydrodistillate, its 19 constituents and 28 structurally related compounds against early third-instar larvae of aedes albopictus (ae. albopictus), aedes aegypti (ae. aegypti) and culex pipiens palles (cx. p. pallens) was examined using direct-contact bioassays. the efficacy of active compounds was further evaluated in semi-field bioassays using field-collected larval cx. p. pallens. results were compared with those of two synthetic larvicides, temephos ...201326464387
high chlorpyrifos resistance in culex pipiens mosquitoes: strong synergy between resistance genes.we investigated the genetic determinism of high chlorpyrifos resistance (hcr), a phenotype first described in 1999 in culex pipiens mosquitoes surviving chlorpyrifos doses ⩾1 mg l(-1) and more recently found in field samples from tunisia, israel or indian ocean islands. through chlorpyrifos selection, we selected several hcr strains that displayed over 10 000-fold resistance. all strains were homozygous for resistant alleles at two main loci: the ace-1 gene, with the resistant ace-1(r) allele ex ...201626463842
aedes aegypti control strategies in brazil: incorporation of new technologies to overcome the persistence of dengue epidemics.dengue is considered to be the most important mosquito-borne viral disease in the world. the aedes aegypti mosquito, its vector, is highly anthropophilic and is very well adapted to urban environments. although several vaccine candidates are in advanced stages of development no licensed dengue vaccine is yet available. as a result, controlling the spread of dengue still requires that mosquitoes be targeted directly. we review the current methods of dengue vector control focusing on recent techni ...201526463204
fighting arbovirus transmission: natural and engineered control of vector competence in aedes mosquitoes.control of aedine mosquito vectors, either by mosquito population reduction or replacement with refractory mosquitoes, may play an essential role in the fight against arboviral diseases. in this review, we will focus on the development and application of biological approaches, both natural or engineered, to limit mosquito vector competence for arboviruses. the study of mosquito antiviral immunity has led to the identification of a number of host response mechanisms and proteins that are required ...201526463078
effects of starvation on deltamethrin tolerance in bed bugs, cimex lectularius l. (hemiptera: cimicidae).bed bugs, cimex lectularius l., are a major pest in the urban environment. their presence often results in physical, psychological, and financial distress of homeowners and apartment dwellers. although many insecticide bioassays have been performed on this pest, little attention has been paid to bed bug feeding status, which is closely linked to metabolism, molting, and mass. therefore, we evaluated the toxicity of topically applied deltamethrin on insecticide susceptible adult male bed bugs fed ...201526463068
bacillus thuringiensis is an environmental pathogen and host-specificity has developed as an adaptation to human-generated ecological niches.bacillus thuringiensis (bt) has been used successfully as a biopesticide for more than 60 years. more recently, genes encoding their toxins have been used to transform plants and other organisms. despite the large amount of research on this bacterium, its true ecology is still a matter of debate, with two major viewpoints dominating: while some understand bt as an insect pathogen, others see it as a saprophytic bacteria from soil. in this context, bt's pathogenicity to other taxa and the possibi ...201326462580
the non-photosynthetic algae helicosporidium spp.: emergence of a novel group of insect pathogens.since the original description of helicosporidium parasiticum in 1921, members of the genus helicosporidium have been reported to infect a wide variety of invertebrates, but their characterization has remained dependent on occasional reports of infection. recently, several new helicosporidium isolates have been successfully maintained in axenic cultures. the ability to produce large quantity of biological material has led to very significant advances in the understanding of helicosporidium biolo ...201326462425
co-circulation of usutu virus and west nile virus in a reed bed ecosystem.mosquito-borne flaviviruses are a major public health threat in many countries worldwide. in central europe, west nile virus (wnv) and usutu virus (usuv), both belonging to the japanese encephalitis virus group (flaviviridae) have emerged in the last decennium. surveillance of mosquito vectors for arboviruses is a sensitive tool to evaluate virus circulation and consequently to estimate the public health risk.201526459018
insights into the recent emergence and expansion of eastern equine encephalitis virus in a new focus in the northern new england usa.eastern equine encephalomyelitis virus (eeev) causes a highly pathogenic zoonosis that circulates in an enzootic cycle involving the ornithophagic mosquito, culiseta melanura, and wild passerine birds in freshwater hardwood swamps in the northeastern u.s. epidemic/epizootic transmission to humans/equines typically occurs towards the end of the transmission season and is generally assumed to be mediated by locally abundant and contiguous mammalophagic "bridge vector" mosquitoes.201526453283
flight capacity of bactrocera dorsalis (diptera: tephritidae) adult females based on flight mill studies and flight muscle ultrastructure.the oriental fruit fly, bactrocera dorsalis (hendel) (diptera: tephritidae), is considered a major economic threat in many regions worldwide. to better comprehend flight capacity of b. dorsalis and its physiological basis, a computer-monitored flight mill was used to study flight capacity of b. dorsalis adult females of various ages, and the changes of its flight muscle ultrastructures were studied by transmission electron microscopy. the flight capacity (both speed and distance) changed signifi ...201526450591
acetylcholinesterases from the disease vectors aedes aegypti and anopheles gambiae: functional characterization and comparisons with vertebrate orthologues.mosquitoes of the anopheles (an.) and aedes (ae.) genus are principal vectors of human diseases including malaria, dengue and yellow fever. insecticide-based vector control is an established and important way of preventing transmission of such infections. currently used insecticides can efficiently control mosquito populations, but there are growing concerns about emerging resistance, off-target toxicity and their ability to alter ecosystems. a potential target for the development of insecticide ...201526447952
evolutionary divergence of core and post-translational circadian clock genes in the pitcher-plant mosquito, wyeomyia smithii.internal circadian (circa, about; dies, day) clocks enable organisms to maintain adaptive timing of their daily behavioral activities and physiological functions. eukaryotic clocks consist of core transcription-translation feedback loops that generate a cycle and post-translational modifiers that maintain that cycle at about 24 h. we use the pitcher-plant mosquito, wyeomyia smithii (subfamily culicini, tribe sabethini), to test whether evolutionary divergence of the circadian clock genes in this ...201526444857
multicopper oxidase-1 is required for iron homeostasis in malpighian tubules of helicoverpa armigera.multicopper oxidases (mcos) are enzymes that contain 10 conserved histidine residues and 1 cysteine residue. mco1 has been extensively investigated in the midgut because this mco is implicated in ascorbate oxidation, iron homeostasis and immune responses. however, information regarding the action of mco1 in malpighian tubules is limited. in this study, helicoverpa armigera was used as a model to investigate the function of mco1 in malpighian tubules. sequence analysis results revealed that hamco ...201526437857
breeding sites of aedes aegypti: potential dengue vectors in dire dawa, east ethiopia.background and objectives. entomological survey was carried out from may-june to september-october 2014 to investigate the presence of dengue vectors in discarded tires and artificial water containers in houses and peridomestic areas. methods. a cross-sectional immature stage survey was done indoors and outdoors in 301 houses. mosquito larval sampling was conducted using pipette or dipper depending on container types. larvae were identified morphologically and larval indices were also calculated ...201526435712
temporal gene expression profiles of pre blood-fed adult females immediately following eclosion in the southern house mosquito culex quinquefasciatus.prior to acquisition of the first host blood meal, the anautogenous mosquito culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. in the current study, we conducted whole transcriptome analyses of adult female culex mosquitoes to identify genes that may be necessary for both taking of the blood meal, and processing of the blood meal in adult female mosquitoes cx. quinquefasciatus. we examined temporal expression of genes for the p ...201526435696
an ace-1 gene duplication resorbs the fitness cost associated with resistance in anopheles gambiae, the main malaria mosquito.widespread resistance to pyrethroids threatens malaria control in africa. consequently, several countries switched to carbamates and organophophates insecticides for indoor residual spraying. however, a mutation in the ace-1 gene conferring resistance to these compounds (ace-1(r) allele), is already present. furthermore, a duplicated allele (ace-1(d)) recently appeared; characterizing its selective advantage is mandatory to evaluate the threat. our data revealed that a unique duplication event, ...201526434951
identification of chemical constituents of zanthoxylum heitzii stem bark and their insecticidal activity against the malaria mosquito anopheles gambiae.zanthoxylum heitzii bark extracts have insecticidal properties and have been reported to be used against malaria in western africa. previously, it has been shown that a hexane extract of the bark is toxic to adult females of the mosquito anopheles gambiae, a malaria vector. as part of our project on the control of malaria vectors using plant extracts, the phytochemistry of z. heitzii bark hexane extract has been investigated with the aim to identify the major components with adulticidal and larv ...201526427956
the wolbachia wo bacteriophage proteome in the aedes albopictus c/wstr1 cell line: evidence for lytic activity?wolbachia pipientis (rickettsiales), an obligate intracellular alphaproteobacterium in insects, manipulates host reproduction to maximize invasion of uninfected insect populations. modification of host population structure has potential applications for control of pest species, particularly if wolbachia can be maintained, manipulated, and genetically engineered in vitro. although wolbachia maintains an obligate mutualism with genome stability in nematodes, arthropods can be co-infected with dist ...201526427709
mosquito age and avian malaria infection.the immune system of many insects wanes dramatically with age, leading to the general prediction that older insects should be more susceptible to infection than their younger counterparts. this prediction is however challenged by numerous studies showing that older insects are more resistant to a range of pathogens. the effect of age on susceptibility to infections is particularly relevant for mosquitoes given their role as vectors of malaria and other diseases. despite this, the effect of mosqu ...201526424326
"singing in the tube"--audiovisual assay of plant oil repellent activity against mosquitoes (culex pipiens).plant essential oils have been suggested as a promising alternative to the established mosquito repellent deet (n,n-diethyl-meta-toluamide). searching for an assay with generally available equipment, we designed a new audiovisual assay of repellent activity against mosquitoes "singing in the tube," testing single mosquitoes in drosophila cultivation tubes. statistics with regression analysis should compensate for limitations of simple hardware. the assay was established with female culex pipiens ...201626412058
mosaic composition of riba and wspb genes flanking the virb8-d4 operon in the wolbachia supergroup b-strain, wstr.the obligate intracellular bacterium, wolbachia pipientis (rickettsiales), is a widespread, vertically transmitted endosymbiont of filarial nematodes and arthropods. in insects, wolbachia modifies reproduction, and in mosquitoes, infection interferes with replication of arboviruses, bacteria and plasmodia. development of wolbachia as a tool to control pest insects will be facilitated by an understanding of molecular events that underlie genetic exchange between wolbachia strains. here, we used n ...201526400107
drosophila americana diapausing females show features typical of young flies.diapause is a period of arrested development which is controlled physiologically, preprogrammed environmentally and characterized by metabolic depression that can occur during any stage of insect development. nevertheless, in the genus drosophila, diapause is almost always associated with the cessation of ovarian development and reproductive activity in adult females. in this work, we show that, in d. americana (a temperate species of the virilis group), diapause is a genetically determined dela ...201526398836
winter activity and aboveground hybridization between the two biotypes of the west nile virus vector culex pipiens.culex (cx.) pipiens mosquitoes are important vectors of west nile virus (wnv). in europe, the species cx. pipiens consists of two biotypes, pipiens and molestus, which are morphologically identical, but differ in behavior. typical behavior of the molestus biotype is the ability to remain active during winter, whereas the pipiens biotype enters diapause. the current paradigm is that the two biotypes occur sympatrically in southern europe, but occur in isolated above- and belowground populations i ...201526394124
early transcriptional events linked to induction of diapause revealed by rnaseq in larvae of drosophilid fly, chymomyza costata.diapause is a developmental alternative to direct ontogeny in many invertebrates. its primary adaptive meaning is to secure survival over unfavourable seasons in a state of developmental arrest usually accompanied by metabolic suppression and enhanced tolerance to environmental stressors. during photoperiodically triggered diapause of insects, the ontogeny is centrally turned off under hormonal control, the molecular details of this transition being poorly understood. using rnaseq technology, we ...201526391666
potential mosquito (diptera:culicidae) vector of dirofilaria repens and dirofilaria immitis in urban areas of eastern slovakia.this paper follows the study from 2013 focused on the molecular screening of mosquitoes as vectors of dirofilaria spp. which provided the information on aedes vexans as a potential vector of dirofilaria repens in slovakia. current entomological and molecular research indicates that ae. vexans can participate also in the transmission of dirofilaria immitis within the region. using the standard pcr method, we examined 10,500 mosquitoes (ae. vexans, ae. rossicus, anopheles maculipennis s.l., ochler ...201526391170
experimental studies on comparison of the vector competence of four italian culex pipiens populations for west nile virus.west nile virus (wnv) is a vector-borne disease responsible for causing epidemics in many areas of the world. the virus is maintained in nature by an enzootic bird-mosquito-bird cycle and occasionally transmitted to other hosts, such as equines and humans. culex species, in particular the ubiquitous species culex pipiens is thought to play a major vector role both in enzootic and epizootic maintenance and transmission of wnv. introduced in europe in recent years, since 2008 wnv has been stably c ...201526383834
evaluation of vector competence for west nile virus in italian stegomyia albopicta (=aedes albopictus) mosquitoes.west nile virus (wnv) is a zoonotic arboviral pathogen transmitted by mosquitoes in a cycle that involves wild birds as reservoir hosts. the virus is responsible for outbreaks of viral encephalitis in humans and horses. in europe, culex pipiens (diptera: culicidae) is considered to be the main vector of wnv, but other species such as stegomyia albopicta (=aedes albopictus) (diptera: culicidae) may also act as competent vectors of this virus. since 2008 human cases of wnv disease have been report ...201526382099
mitochondrial genomes of two phlebotomine sand flies, phlebotomus chinensis and phlebotomus papatasi (diptera: nematocera), the first representatives from the family psychodidae.leishmaniasis is a worldwide but neglected disease of humans and animal transmitted by sand flies, vectors that also transmit other important diseases. mitochondrial genomes contain abundant information for population genetic and phylogenetic studies, important in disease management. however, the available mitochondrial sequences of these crucial vectors are limited, emphasizing the need for developing more mitochondrial genetic markers.201526381614
comparative transcriptome analyses of deltamethrin-susceptible and -resistant culex pipiens pallens by rna-seq.the widespread and improper use of pyrethroid insecticides, such as deltamethrin, has resulted in the evolution of resistance in many mosquito species, including culex pipiens pallens. with the development of high-throughput sequencing, it is possible to massively screen pyrethroid resistance-associated gene. in this study, we used illumina-solexa transcriptome sequencing to identify genes that are expressed differently in deltamethrin-susceptible and -resistant strains of culex pipiens pallens ...201626377942
limited genomic divergence between intraspecific forms of culex pipiens under different ecological pressures.divergent selection can be a major driver of ecological speciation. in insects of medical importance, understanding the speciation process is both of academic interest and public health importance. in the west nile virus vector culex pipiens, intraspecific pipiens and molestus forms vary in ecological and physiological traits. populations of each form appear to share recent common ancestry but patterns of genetic differentiation across the genome remain unknown. here, we undertook an aflp genome ...201526377220
modification of the suna trap for improved survival and quality of mosquitoes in support of epidemiological studies.monitoring adult mosquito populations provides information that is critical for assessing risk of vector-borne disease transmission. the recently developed suna trap was found to be a very effective trap when baited with an attractive odor blend. a modification of this trap was tested to improve its function as a tool for monitoring mosquito populations, including anopheles coluzzii (an. gambiae sensu stricto molecular form m), aedes aegypti, and culex pipiens. the modified suna trap (suna-m) wa ...201526375903
wolbachia strains for disease control: ecological and evolutionary considerations.wolbachia are endosymbionts found in many insects with the potential to suppress vectorborne diseases, particularly through interfering with pathogen transmission. wolbachia strains are highly variable in their effects on hosts, raising the issue of which attributes should be selected to ensure that the best strains are developed for disease control. this depends on their ability to suppress viral transmission, invade host populations, persist without loss of viral suppression and not interfere ...201526366194
larval habitats diversity and distribution of the mosquito (diptera: culicidae) species in the republic of moldova.a countrywide field survey of immature mosquitoes was conducted in moldova with the aim to evaluate the culicidae species composition in different larval habitats and their distribution in the country. in total, 259 potential larval habitats were sampled in the 53 localities, resulting in 9,456 specimens. twenty species belonging to the genera anopheles, aedes, culex, culiseta, and uranotaenia were collected. mean species richness in aquatic habitats ranged from 1.00 to 4.00, and, for example, w ...201526364191
mosquitocidal properties of bacillus species isolated from mangroves of vellar estuary, southeast coast of india.samples collected from the mangroves of vellar estuary yielded a mosquitocidal bacterium, whose secondary metabolites exhibited mosquito larvicidal and pupicidal activity. the bacterium was isolated using standard microbiological methods and identified using classical biochemical tests. the mosquitocidal bacterium was identified as bacillus subtilis, bacillus thuringiensis, bacillus sphaericus and bacillus cereus. mosquitocidal metabolite(s) was separated from the culture supernatant of the bact ...201326345039
additive effects on the improvement of insecticidal activity: design, synthesis, and insecticidal activity of novel pymetrozine derivatives.a series of new pymetrozine analogues containing both methyl on the imine carbon and phenoxy group at the pyridine ring were designed and synthesized. their insecticidal activities against bean aphid (aphis craccivora), mosquito larvae (culex pipiens pallens), cotton bollworm (helicoverpa armigera), corn borer (ostrinia nubilalis) and oriental armyworm (mythimna separata) were evaluated. the results of bioassays indicated that most of the target compounds showed good insecticidal activity agains ...201626342545
bagaza virus is pathogenic and transmitted by direct contact in experimentally infected partridges, but is not infectious in house sparrows and adult mice.bagaza virus (bagv) is a mosquito-borne flavivirus belonging to the ntaya serocomplex. in 2010, a disease outbreak was reported in cádiz (southern spain) affecting game birds (red-legged partridges and common pheasants). in this work, red-legged partridges were inoculated experimentally with infectious bagv isolated from this outbreak in order to make a complete clinical and analytical assessment of the disease caused by the pathogen in this species. viral load (by real-time rt-pcr) in blood, or ...201526338714
the cuticle proteins: a putative role for deltamethrin resistance in culex pipiens pallens.insecticide resistance has been a major public health challenge. it is impendent to study the mechanism on insecticide resistance. in our previous study, 14 differentially accumulated insect cuticle proteins (icps) based on insecticide resistance proteomes and transcriptomes were found in the deltamethrin-resistant (dr) and -susceptible (ds) strains of culex pipiens pallens. to investigate if these icps are associated with deltamethrin resistance, different transcriptional levels of the 14 icps ...201526337265
synthesis and antiviral, insecticidal, and fungicidal activities of gossypol derivatives containing alkylimine, oxime or hydrazine moiety.gossypol is a part of the cotton plant's defense system against pathogens and herbivorous insects. to discover gossypol analogs with broad spectrum and high activity, a series of gossypol alkylamine schiff base, oxime and hydrazone derivatives were synthesised and bioassayed. the biological results indicated that most of these derivatives exhibited higher anti-tmv activity than gossypol. interestingly, the activities of compounds 10, 15, 18, 20, 23 and 26 were much higher than that of ribavirin. ...201626337022
mom matters: diapause characteristics of culex pipiens-culex quinquefasciatus (diptera: culicidae) hybrid mosquitoes.females of the northern house mosquito, culex pipiens l., are capable of entering an adult overwintering diapause characterized by arrested ovarian development, enhanced stress tolerance, and elevated lipid stores. in contrast, the southern house mosquito, culex quinquefasciatus say, lacks this capacity and is therefore unable to survive the harsh winters found in northern regions of north america. these two species are capable of forming fertile hybrids in the united states, yet the diapause ch ...201526336296
seasonal and daily activity patterns of mosquito (diptera: culicidae) vectors of pathogens in northeastern italy.the seasonal and daily activity of mosquito vectors of pathogens affecting animals and humans were studied in northeastern italy at a site within the po river delta park. a cdc-co2 trap and a gravid trap were operated at 2-h intervals for 24 h every 15 d from may to october 2010. overall, 5,788 mosquitoes comprising six species were collected, namely culex pipiens l. (75.1% of total), aedes caspius (pallas) (15.2%), aedes vexans (meigen) (6.9%), anopheles maculipennis s.l. meigen (2.6%), culiset ...201526336280
simulated seasonal photoperiods and fluctuating temperatures have limited effects on blood feeding and life history in aedes triseriatus (diptera: culicidae).biotic and abiotic factors change seasonally and impact life history in temperate-zone ectotherms. temperature and photoperiod are factors that change in predictable ways. most studies testing for effects of temperature on vectors use constant temperatures and ignore potential correlated effects of photoperiod. in two experiments, we tested for effects of larval rearing environments creating ecologically relevant temperatures and photoperiods simulating early and late season conditions (june and ...201526336255
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