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Host preference of mosquito vectors of Japanese encephalitis.

The host preference of 4 Culex mosquito species collected in Miaoli and Pingtung counties, Taiwan was studied by capillary precipitin method. Antisera to alum-precipitated sera of man, bovine, swine, rabbit, horse, dog, cat, mouse, chicken, duck, and pigeon were produced in rabbits and reacted with 758 mosquito blood meals among which reactions to one or more antisera. Culex annulus and Culex tritaeniorhynchus summorosus showed a great avidity for pig, and Culex fuscocephala for bovine. Culex pipiens fatigans was ornithophilic. None of 110 C. t. summorosus and 2.4% of 223 C. annulus had fed on man. Among 66 samples of C.p. fatigans tested 10.3% had fed on man, while none of 359 C. fuscocephala did. It seems that the latter does not act as a primary vector of Japanese encephalitis.

Animals

Epidemiological studies on Japanese encephalitis in Kyoto City area, Japan. I. Evidence for decrease of vector mosquitoes.

Mosquito collections by using light traps have been carried out at 10 to 11 stations in Kyoto City area at intervals of about 10 days every year. Mean percent indexes (MPI), being calculated from the data of mosquito collections, were used for comparison of the annual abundance of mosquitoes. It is no doubt that Culex tritaeniorhynchus summorosus has decreased recently and this decrease is correlated with the reduction of human patients of Japanese encephalitis. Wide use of two herbicides, CNP and nitrofen, for rice plant cultivation, may probably be one of the reasons for the decrease of the mosquitoes.

Animals

Genetic analysis of two enzyme polymorphisms in a malaria vector mosquito: octanol dehydrogenase and acid phosphatase in Anopheles culicifacies Giles.

Genetic analysis was performed on two polymorphic enzyme systems in the malaria vector, Anopheles culicifacies Giles. The data indicate that both enzymes, octanol dehydrogenase and acid phosphatase, are controlled by autosomal loci but that these two loci are not linked. The three expected linkage groups in this mosquito have now been identified: linkage group I contains sex and rose-eye; linkage group II contains Odh; and linkage group III contains Acph.

Acid Phosphatase

Dirofilaria immitis and its potential mosquito vectors in central New York State.

Screening of 25,822 dog blood samples indicated approximately a 1% infection rate with Dirofilaria immitis. Dipetalonema reconditum microfilariae were found in approximately 2% of 1,876 feral dogs examined. Laboratory experimentation indicated that 6 of 10 local mosquito species examined allowed successful extrinsic incubation of D immitis. Indices of experimental infection indicated Aedes triseriatus and Anopheles quadrimaculatus were excellent hosts for D immitis, but other factors considered, Aedes vexans probably served as the primary vector of D immitis. Coquillettidia perturbans and Culex pipiens-restuans group were considered unsuitable hosts and thus unlikely vectors of canine heartworm. Natural filarial infections were found in An quadrimaculatus and Ae vexans.

Aedes

Epidemiological studies on Japanese encephalitis in Kyoto City area, Japan. III. Seasonal prevalence of virus infections in several pig populations shown by virus recovery from engorged Culex tritaeniorhynchus summorosus.

When vector mosquitoes engorged by feeding on pigs are tested for virus recovery after incubation for 7 to 10 days, the results may show mosquito infection itself. Therefore, seasonal prevalence of infection in each pig population can be estimated from course of the infection rate among mosquitoes. Many mosquitoes of the main vector of Japanese encephalitis virus, Culex tritaeniorhynchus summorosus, collected by light traps everyday or every other day in some pig sheds from 1967 to 1970 were tested for virus recovery after incubation. The tests were positive during about a month period each summer, and the peak infection rate was high being over 10%. The course of the mosquito infection showed a certain pattern with one or two peaks between the initial recovery and the highest peak. From the interval of 12 to 13 days after the first recovery to the peak, cyclic infection between the pig and the mosquito may occur at the same interval.

Animals

Protocol for Detecting and Sequencing Chikungunya Virus from Field-Collected Mosquitoes.

Arboviral diseases represent a major public health challenge, especially in tropical regions where environmental conditions may favor the proliferation and spread of mosquito vectors. Thus, early and accurate detection of chikungunya virus (CHIKV) in mosquito populations can be a valuable tool for effective surveillance of circulating variants and for identifying new viral introductions. Given the challenges of detecting arboviruses in field-captured mosquitoes, we describe an integrated workflow for CHIKV molecular detection and whole-genome sequencing. This protocol includes mosquito homogenization using a bead-based mechanical disruptor, RNA extraction using TRIzol reagent with minor modifications, molecular screening using CHIKV-specific RT-qPCR, and whole-genome amplification followed by sequencing on Illumina platforms. Despite the protocol being optimized for individual mosquitoes, it results in high-quality RNA suitable for both entomological surveillance and genomic analysis. As this protocol allows recovery of complete CHIKV genomes from mosquito specimens, it can serve as a basis for genomic epidemiology studies, enabling monitoring of viral diversity and lineage dynamics, and facilitating early detection of emerging variants to support timely and targeted public health interventions in endemic and at-risk regions.

Animals

Insecticide susceptibility of some vector fleas and mosquitoes in Burma.

Rat fleas and mosquitoes are insect vectors of public health importance in Burma. Plague is endemic in Central Burma and DDT has been the principal insecticide used for its control to date. Dengue haemorrhagic fever, recently introduced and transmitted by Aedes aegypti, has been spreading to major towns since 1971. The rodents, Rattus rattus, R. exulans, Bandicota bengalensis, Mus musculus, as well as shrews were commonly caught during routine trapping in the country. Rattus norvegicus, prevalent in Rangoon City, is not found in Central Burma. The rat fleas, Xenopsylla cheopis and X. astia, were found to be infesting these rodents, the former being the principal vector of plague. Insecticide susceptibility tests have been carried out periodically in plague endemic areas and Rangoon since 1966 and it is now observed that rat fleas in most of these endemic towns and Rangoon Port are no longer susceptible to DDT. Subsequent rechecks in towns where fleas became resistance to DDT show that fleas are persistently resistant to the insecticide. Aedes aegypti is found to be highly prevalent both in rural and urban areas of almost every major town and townships below 900 meters. Insecticide susceptible tests on this mosquito in limited areas show that the mosquito is generally resistant to DDT but susceptible to other insecticides. With the development of DDT resistance in fleas, it is now necessary to change to an effective insecticide in the control of plague in Burma.

Aedes

Expansive and Diverse Phenotypic Landscape of Field Aedes aegypti (Diptera: Culicidae) Larvae with Differential Susceptibility to Temephos: Beyond Metabolic Detoxification.

Arboviruses including dengue, Zika, and chikungunya are amongst the most significant public health concerns worldwide. Arbovirus control relies on the use of insecticides to control the vector mosquito Aedes aegypti (Linnaeus), the success of which is threatened by widespread insecticide resistance. The work presented here profiled the gene expression of Ae. aegypti larvae from field populations of Ae. aegypti with differential susceptibility to temephos originating from two Colombian urban locations, Bello and Cúcuta, previously reported to have distinctive disease incidence, socioeconomics, and climate. We demonstrated that an exclusive field-to-lab (Ae. aegypti strain New Orleans) comparison generates an over estimation of differential gene expression (DGE) and that the inclusion of a geographically relevant field control yields a more discrete, and likely, more specific set of genes. The composition of the obtained DGE profiles is varied, with commonly reported resistance associated genes including detoxifying enzymes having only a small representation. We identify cuticle biosynthesis, ion exchange homeostasis, an extensive number of long noncoding RNAs, and chromatin modelling among the differentially expressed genes in field resistant Ae. aegypti larvae. It was also shown that temephos resistant larvae undertake further gene expression responses when temporarily exposed to temephos. The results from the sampling triangulation approach here contribute a discrete DGE profiling with reduced noise that permitted the observation of a greater gene diversity, increasing the number of potential targets for the control of insecticide resistant mosquitoes and widening our knowledge base on the complex phenotypic network of the Ae. aegypti response to insecticides.

Aedes

Equine viral encephalitis.

The most important neurotropic viral infections of the horse are the arthropod-borne encephalitides. These include Venezuelan encephalitis (VE), eastern encephalitis (EE) and western encephalitis (WE), which are found in the Americas, and Japanese B encephalitis which occurs in the Far East. All the viruses cause encephalitis in man. Between 1969 and 1972 an epidemic of VE occurred in Central America. In 1971 the disease was reported in Texas, where it was brought under control by the vaccination of susceptible horses with an attenuated live virus vaccine and by the reduction of the mosquito population with insecticides sprayed from aircraft. A high titre viraemia occurs with VE virus in the horse and epidemics are maintained by a mosquito/horse cycle; infection of man and other species is incidental. EE and WE have been recognised as separate diseases since 1933 and in the U.S.A. horses are protected by routine vaccination. Epidemics of these diseases are routine vaccination. Epidemics of these diseases are now uncommon. In contrast with VE, both EE and WE viruses are maintained by a bird/mosquito cycle. The viraemia in the horse is generally considered insufficient to infect mosquito vectors; the horse is a "dead end host". Several species of mosquito can act as vectors of VE, WE and EE. The extension of other arthropod-borne diseases to areas originally outside their geographical distribution (e.g. bluetongue in sheep) serves to illustrate the potential of VE, WE and EE to cause disease on other continents.

Animals

Autodissemination stations suppress Aedes notoscriptus mosquitoes and reduce Buruli ulcer risk in urban Australia: a randomized controlled field trial.

Aedes notoscriptus are mosquito vectors implicated in transmission of Mycobacterium ulcerans. This bacterium causes a destructive infection of skin and soft tissue called Buruli ulcer. Here we ran a randomized controlled trial in an urban Buruli ulcer endemic area in Melbourne, Australia to test whether autodissemination mosquito control stations, containing pyriproxyfen (larvicide) and Beauveria bassiana (entomopathogenic fungus), suppress Ae. notoscriptus populations. Six geographic areas each received 100 autodissemination stations for 8 weeks, and six control areas received no stations between 25 January 2024 and 21 March 2024. The primary outcome measure was mosquito population numbers. After the trial, there was a 70% average reduction in mosquito egg counts among the six intervention areas compared to control areas (P = 0.0076). In an ad hoc analysis, we then explored human Buruli ulcer notifications in treatment and control areas. After accounting for the 4.8-month mean incubation period, there was an 83% reduction in infection likelihood coinciding with peak intervention effect (intervention zones 1 case, control zones 6 cases, incidence ratio rate 0.167, 95% CI 0.0026-1.054, P = 0.047). The effect was not observed during the same time period in the year previous or following 2024, when no interventions were undertaken. A strong correlation (R2 = 0.85) was observed between decreased disease risk and mosquito suppression. These data show that autodissemination traps can effectively lower urban mosquito populations and reduce the threat of Buruli ulcer in humans.

Animals

Recent advances in applied malaria immunology.

Our present knowledge of cellular and humoral factors which are involved in immunity to plasmodial infections are discussed. Immunization against plasmodial infection has been achieved in birds, rodents, simians, and humans. Avian hosts have been immunized against gametocytes which resulted in inhibition of gametocytes within the mosquito vector. Immunization of humans against plasmodial gametocytes would indirectly protect them against malaria by blocking mosquito transmission to other susceptible individuals. Immunization by sporozoites provides short-lived protection against sporozoite challenge, but gives no protection against erythrocytic forms. Some success has been obtained in immunizing avian and mammalian hosts with exoerythrocytic forms obtained from cultured avian cells. The most significant advances have occurred in immunizing simian hosts against simian or human malaria by vaccinating with fresh erythrocytic merozoites or a nonviable lyophilized antigen obtained from intraerythrocytic forms. The development of an antigen preparation suitable for use as a human malaria vaccine is dependent upon prior development of an in vitro system which would provide adequate amounts of parasite material. Efforts to cultivate the sporogonic, exoerythrocytic, and erythrocytic, and erythrocytic phases of plasmodia as well as the feasibility of using these forms for vaccination are discussed.

Animals

Altered histone modifications in Aedes aegypti following Rift Valley fever virus exposure.

When arthropod-borne viruses (arboviruses) are delivered to vector mosquitoes in an infectious bloodmeal, viral components interact with host proteins to hijack cells and initiate replication. The extent to which arbovirus infection alters mosquito host transcriptional and genomic regulatory processes is currently unknown. We hypothesized that histone modifications would be altered in mosquitoes exposed to Rift Valley fever virus (RVFV MP12, Phlebovirus riftense, family Phleboviridae). We interrogated transcriptome and chromatin landscapes in Aedes aegypti midguts by performing Cleavage Under Targets and Release Using Nuclease (CUT&RUN), using H3K27ac and H3K9me3 marks. Altered H3K27ac marks were identified following RVFV MP12 exposure, as well as upon bloodfeeding alone. It took several days for differential H3K27ac marks to be associated with differentially expressed genes (DEGs) in RVFV-exposed midguts. H3K27ac peaks showed progressive depletion as infection progressed. Gene set enrichment analysis revealed that immune response transcripts were enriched at 1 and 3 dpf (days post-feeding) but depleted by 7 dpf. Hedgehog/Gli (glioma-associated oncogene homolog) signaling pathway transcripts were depleted, indicating possible viral manipulation of cellular polarization. Moreover, at 7 dpf, 7 of 102 DEGs were proximal to differentially acetylated sites in a pattern expected to favor viral propagation. However, one transcript coding for an antiviral effector (LysM-TLDc domain protein) showed significant depletion of both H3K9me3 and H3K27ac marks. Analysis of midguts after a non-infectious bloodmeal versus sugar-fed controls revealed global changes to H3K27ac and H3K9me3 marks during and following the period of bloodmeal digestion. Differential H3K27ac marks were proximal to one quarter of all DEGs at 1 dpf, consistent with an important role of H3K27ac in bloodmeal digestion. These results demonstrate that H3K27ac and H3K9me3 patterns are altered upon virus exposure in a complex interplay that favors viral replication but is also countered by host responses to limit replication.

ChIP-Seq

Gamete vaccines and transmission-blocking immunity in malaria.

We have recently proposed an approach to malaria control based on immunization of the host against extracellular malarial gametes, the stage in the mosquito guts, in order to block transmission by the mosquito vector. Our studies with avian and primate models have demonstrated that immunization of the host with extracellular gametes totally suppresses infectivity to the mosquito of a subsequent blood meal. Gametocytes within the erythrocytes are unaffected by the immunity, since resuspending the gametocytes in serum from normal nonimmune animals restores their infectivity to mosquitos. Immunity is mediated by antibodies that are ingested with the blood meal. These antibodies interact with extracellular gametes and prevent fertilization (the fusion of male and female gametes). Thus the infection in the mosquito is blocked, and in this way transmission is interrupted.

Animals