Comparative Study of the Pathological Effects of Western Equine Encephalomyelitis Virus in Four Strains of Culex tarsalis Coquillett (Diptera: Culicidae).

Early reports suggested that mosquito cells infected with arboviruses remain viable and undamaged. However, more recent experimental evidence suggests that arboviral infection of mosquito tissues might indeed result in pathological changes, with potential implications for vector survival and virus transmission. Here, we compare the pathological effects of western equine encephalomyelitis virus (WEEV) infection in four strains of Culex tarsalis previously reported to differ in their competence as WEEV vectors.

Infection dynamics of western equine encephalomyelitis virus (Togaviridae: Alphavirus) in four strains of Culex tarsalis (Diptera: Culicidae): an immunocytochemical study.

BACKGROUND: Vector competence describes the efficiency with which vector arthropods become infected with and transmit pathogens and depends on interactions between pathogen and arthropod genetics as well as environmental factors. For arbovirus transmission, the female mosquito ingests viremic blood, the virus infects and replicates in midgut cells, escapes from the midgut, and disseminates to other tissues, including the salivary glands. Virus-laden saliva is then injected into a new host.

Rift Valley fever virus-infected mosquito ova and associated pathology: possible implications for endemic maintenance.

Background: Endemic/Enzootic maintenance mechanisms like vertical transmission (pathogen passage from infected adults to their offspring) are central in the epidemiology of zoonotic pathogens. In Kenya, Rift Valley fever virus (RVFV) may be maintained by vertical transmission in ground-pool mosquitoes such as . RVFV can cause serious morbidity and mortality in humans and livestock.

Meconial peritrophic matrix structure, formation, and meconial degeneration in mosquito pupae/pharate adults: histological and ultrastructural aspects.

The noncellular peritrophic matrix (PM) that forms around the food bolus in the midgut of many arthropod species may influence the fate of ingested microbes. In mosquitoes, PMs have been identified in the pupal as well as larval and adult stages. In pupae, the PMs surround the meconium, the sloughed larval midgut epithelium.

Pathogenesis of Rift Valley fever virus in mosquitoes--tracheal conduits & the basal lamina as an extra-cellular barrier.

Knowledge of the fate of an arbovirus in a mosquito is fundamental to understanding the mosquito's competence to transmit the virus. When a competent mosquito ingests viremic vertebrate blood, virus infects midgut epithelial cells and replicates, then disseminates to other tissues, including salivary glands and/or ovaries. The virus is then transmitted to the next vertebrate host horizontally via bite and/or vertically to the mosquito's offspring.

Evidence for arbovirus dissemination conduits from the mosquito (Diptera: Culicidae) midgut.

The mechanism by which arboviruses bypass the basal lamina of mosquito midgut cells and enter the body cavity has been unclear. Experiments using Venezuelan equine encephalitis viral replicon particles, which express the green fluorescent protein gene in cells, indicate the operation of tissue conduits, possibly involving tracheae and visceral muscles, that facilitate virus movement through the basal lamina. Ultrastructural studies of the midgut reveal evidence for possible complete penetration of the basal lamina by tracheal cells and regions of modified basal lamina associated with visceral muscle.

The energetic costs of diving in Aedes aegypti and Aedes albopictus pupae.

Undisturbed mosquito pupae rest at the water surface and respond to passing shadows or vibrations by diving. Pupae do not feed and rely solely on energy stored from the larval stage. The ability of a newly emerged adult mosquito to survive, and therefore to transmit disease, depends on these energy reserves.

Meconial peritrophic membranes and the fate of midgut bacteria during mosquito (Diptera: Culicidae) metamorphosis.

The location of midgut bacteria relative to meconial peritrophic membranes (MPMs) and changes in bacterial numbers during midgut metamorphosis were studied in Anopheles punctipennis (Say), Culex pipiens (L.), and Aedes aegypti (L.) pupae and newly emerged adults.

The occurrence and fate of the meconium and meconial peritrophic membranes in pupal and adult mosquitoes (Diptera: Culicidae).

In mosquitoes, in addition to larval and adult peritrophic membranes (PMs), a PM (meconial peritrophic membrane or MPM1) forms in the pupa around the meconium, the sloughed, degenerating larval midgut epithelium. Often, a second membrane (MPM2) forms in temporal proximity to adult emergence. Differences in the occurrence, persistence, and timing of disappearance of the meconium/MPMs and gas were studied by dissecting the midgut contents from pupae of known ages postpupation and from adults of known ages postemergence.

Buoyancy and diving behavior in mosquito pupae.

Mosquito pupal diving behavior has been studied mostly in Aedes aegypti and in this species pupal buoyancy varies relative to several factors. The research reported herein addresses the 2 following questions. Does diving behavior vary among different mosquito genera and species? How is diving behavior influenced by variation in buoyancy? Depth and duration of dive, and dive pattern, were compared among Ae.

Rift Valley fever virus in the cardia of Culex pipiens: an immunocytochemical and ultrastructural study.

In this paper, we consider the movement of Rift Valley fever (RVF) virus from infected mosquito midgut epithelial cells into the hemocoel as an important factor in the ultimate ability of the insect to transmit the virus. Our results are therefore significant in the context of vector competence. The mosquito Culex pipiens was identified as the primary vector of RVF in an epidemic that occurred in Egypt in the 1970s.

Effect of the developmental stage at infection on the ability of adult Anopheles stephensi to transmit Rift Valley fever virus.

The ability of adult Anopheles stephensi to transmit Rift Valley fever virus was determined for mosquitoes inoculated at selected times during development. None of 109 female An. stephensi inoculated as adults transmitted virus to hamsters.

Effect of mechanical shock on hydrostatic balance and survival of mosquito pupae.

The presence and distribution of ventral air space gas assures that mosquito pupae are positively buoyant and that they float, dive, and ascend in an upright, balanced orientation. Our objective was to test the effects of mechanical shocks of varying magnitude on mosquito pupae representing 3 genera. Forces that disrupt the pupa's buoyancy and/or hydrostatic balance are of a much lower magnitude than those that would cause tissue damage.

An immunocytochemical study of the distribution of Rift Valley fever virus in the mosquito Culex pipiens.

The dissemination of Rift Valley fever (RVF) virus in females of the epidemic vector Culex pipiens was studied immunocytochemically. Among orally infected specimens, viral antigen was detected in all major midgut regions, although individuals varied with respect to which regions were infected. Among specimens with disseminated infections (infections beyond the midgut epithelium), antigen was detected in most tissues, including those of the nervous and endocrine systems.

Impact of stressful conditions on the survival of Culex pipiens exposed to Rift Valley fever virus.

Several groups of Rift Valley fever (RVF) virus-exposed and unexposed Culex pipiens were allowed differential access to a carbohydrate food source and their survival monitored. When stressed by deprivation of a carbohydrate source, mean survival times of RVF virus-exposed mosquitoes were consistently higher than those of unexposed mosquitoes in each of the carbohydrate-deficient experiments. These differences were statistically significant when mosquitoes were provided 5% sucrose for 24 hours.

Histological parameters useful in the identification of multiple bloodmeals in mosquitoes.

Culex nigripalpus females were given double and, in 2 cases, triple (interrupted) bloodmeals separated by various intervals. Mosquitoes given single meals served as controls. Using Azan-stained serial paraffin sections, we could identify multiple meals separated by 1-72 hr in 44 of 53 cases (84.

The distribution of Rift Valley fever virus in the mosquito Culex pipiens as revealed by viral titration of dissected organs and tissues.

Distribution of Rift Valley fever virus (RVFV) was studied in the mosquito Culex pipiens. Mosquitoes were dissected on days 1-7 after an infectious bloodmeal, and RVFV plaque assays were performed on the legs, posterior midgut, ovaries, salivary glands, thoracic ganglia, and remaining organs and tissues (remnants). On days 7-12 and 14 following an infectious bloodmeal, mosquitoes were tested for their ability to transmit virus and then dissected.

Togavirus-associated pathologic changes in the midgut of a natural mosquito vector.

Arthropod-borne viruses were not previously believed to cause discernible pathologic changes in their natural mosquito vectors. We report cytopathologic lesions in the midgut of the mosquito, Culiseta melanura, 2 to 5 days after oral infection with eastern equine encephalomyelitis virus. Sloughing of densely staining, heavily infected epithelial cells into the midgut lumen was observed by light and transmission electron microscopy, along with degeneration of cells within the epithelium.

Reduced survival of adult Culex pipiens infected with Rift Valley fever virus.

The effect of Rift Valley fever (RVF) viral infection on the survival of female Culex pipiens was examined. In 3 experiments in which mosquitoes ingested RVF virus, there was a 44% decrease in survival to days 14-16 for transmitting vs. nontransmitting mosquitoes, and a 48% decrease in survival for individuals with disseminated vs.

The origin and movement of gas during adult emergence in Aedes aegypti: an hypothesis.

Several events which precede adult emergence in Aedes aegypti were studied, including changes in buoyancy, changes visible through the pupal cuticle and changes at the foregut-midgut junction. Our data suggest tht the gas which is present in the posterior midgut at the time of emergence originates in the tracheal system. During the process of emergence this gas moves into the exuvial space through the adult spiracles and then follows the exuvial fluid into the alimentary canal.

Proteolytic activity in the ectoperitrophic fluid of blood-fed Culex nigripalpus.

In blood-fed Culex nigripalpus Theobald, proteolytic activity appeared in the ectoperitrophic fluid after 3 h, but only after 6 h in a homogenate of the blood-filled midgut. The activity continued to be higher in ectoperitrophic fluid than in whole gut homogenate until about 40 h after the meal, when most of the intact clot had disappeared. Apparently, undigested blood inhibits proteolytic activity.

A behavioral mechanism for resting site selection by pupae in three mosquito species.

Pupae of Aedes aegypti, Ae. triseriatus and Culex restuans dive less frequently when resting in a concave meniscus than when resting in open water. They also tend to terminate diving after contacting submerged vertical surfaces, increasing their chances of surfacing in a concave meniscus.

Use of the avidin-biotin-peroxidase complex immunocytochemical procedure for detection of Rift Valley fever virus in paraffin sections of mosquitoes.

The avidin-biotin-peroxidase complex (ABC) immunocytochemical procedure was used to locate Rift Valley fever (RVF) virus antigen in infected Aedes albopictus C6/36 cultured cells and in serial paraffin sections of intrathoracically-injected Egyptian Culex pipiens. Fixation of the cultured cells in formaldehyde or periodate-lysine-paraformaldehyde, and fixation of whole mosquitoes with formaldehyde resulted in good preservation of morphology and excellent differential staining between uninfected and infected specimens. Two primary antibodies against RVF virus were tested on the cultured cells: a polyclonal rabbit antiserum and a mixture of mouse monoclonal antibodies.