Transmission of Rift Valley fever virus by adult mosquitoes after ingestion of virus as larvae

We studied the ability of Culex pipiens, Aedes circumluteolus, and Ae. mcintoshi, exposed as larvae to liver tissue from a Rift Valley fever (RVF) virus-infected hamster, to become infected and transstadially transmit virus to the adult and for the adults to transmit virus by bite to hamsters. After exposure as larvae, 9% (5/54) of adult Cx. pipiens and 8% (38/505) of the adult Ae. (Neomelaniconion) species were infected. All of the infected Cx. pipiens and about half of the infected Ae. circumluteolus and Ae. mcintoshi tested transmitted RVF virus by bite to hamsters. Transmission rates for mosquitoes orally infected as larvae were higher than those for mosquitoes orally infected as adults. Animals infected with RVF virus may abort or die in the vicinity of mosquito larvae breeding habitats and infected tissue from these animals may contaminate the water.     

Reduced Rift Valley fever virus infection rates in mosquitoes associated with pledget feedings

Infection rates were compared in Culex pipiens and Aedes taeniorhynchus after they fed on Rift Valley fever (RVF) viremic hamsters or ingested similar doses of RVF virus from blood-soaked pledgets. Infection rates were significantly lower for mosquitoes that ingested virus from a pledget than for those that ingested similar doses from viremic hamsters. The method used to prevent normal clot formation for the pledget feedings (i.e., defibrination by shaking with glass beads or addition of heparin) did not affect subsequent infection rates. Both inhibition of normal clot formation and freezing of virus after it had last been propagated were associated with significantly reduced infection rates with the pledget feedings. Laboratory studies using artificial feeding techniques may not give reliable estimates of the vector competence of mosquitoes for arboviruses.     

A comparision of West Nile Virus transmission by Ochlerotatus trivittatus (COQ.), Culex pipiens (L.), and Aedes albopictus (Skuse)

Transmission of West Nile virus (WNV) by Ochlerotatus trivittatus, Culex pipiens, and Aedes albopictus were compared 14 days after taking blood meals from viremic chickens with titers ranging from 10(2.5) to 10(9.5) cell infective dose (50)s (CID50s)/mL serum. Transmission occurred in one of four (25%) Oc. trivittatus and one of 25 (4%) Cx. pipiens that fed on chickens with titers of 10(5.5) CID50s/mL. No transmission occurred among two of 16 (13%) Oc. trivittatus or one of 25 (4%) Cx. pipiens that became infected after blood meals with titers of 10(5.0) and 10(4.5) CID50s/mL, the next lowest blood meal titers evaluated. Seventeen of 28 (61%) Ae. albopictus transmitted WNV after blood meals with titers of 10(7.0) CID50s/mL, but no infection or transmission was observed among 21 Ae. albopictus that fed on chickens with titers of 10(5.0) CID50s/mL, the next lowest titer evaluated. Transmission by all three species increased dramatically after blood meals with WNV titers of > or = 10(5.5) CID50s/mL. No significant differences occurred in dissemination and transmission rates of the three species after taking blood meals with titers of > 10(7.0) CID50s/mL. The cumulative mean +/- SE transmission rates of Oc. trivittatus, Cx. pipiens, and Ae. albopictus after blood meals with titers of > or = 10(7.0) CID50s/mL were 45.5 +/- 4.1%, 46.8 +/- 4.5%, and 72.4 +/- 5.5%. The cumulative mean dissemination rates of the three species were 78.3 +/- 6.7%, 74.8 +/- 2.6%, and 88.6 +/- 2.1%. The rates of transmission by the three species that developed disseminated infections after blood meals with titers of > or = 10(7.0) CID50s/mL were 58.8 +/- 4.4%, 62.6 +/- 5.8%, and 81.6 +/- 5.4%, respectively. In a previous study, we found that susceptibility of the three species to WNV was essentially the same when fed on chickens with WNV titers of > 10(7.0) CID50s/mL, but Oc. trivittatus and Cx. pipiens were more susceptible than Ae. albopictus to WNV at lower virus titers. The current study strongly suggests that Ae. albopictus is a more efficient vector than Oc. trivittatus and Cx. pipiens when fed blood meals with titers of > 10(7.0) CID50s/mL. However, Oc. trivittatus and Cx. pipiens might be more efficient as vectors when infected by blood meals with titers of < 10(7.0) CID50s/mL.     

Increased mosquito feeding on Rift Valley fever virus-infected lambs

Experiments were conducted to determine whether or not mosquitoes exhibited an increase in feeding on Rift Valley fever (RVF) virus-infected lambs as compared to uninfected ones. Overall, when given a choice between a viremic lamb, or its uninfected twin, 65% (1,264/1,943) of the engorged female Culex pipiens obtained blood from the viremic lamb. This excess of feedings on viremic lambs was demonstrated regardless of the age of the lambs, or the time period between inoculation of the lambs and exposure to mosquitoes. With 3-day-old and 6- to 8-week-old lambs, 66% of the mosquitoes obtained blood from the viremic lamb. At 28, 48, and 52 hours after infection, the percentages of mosquitoes feeding on viremic lambs were 72, 68, and 64, respectively. Temperature, as measured by the difference in temperature of the viremic lamb minus that of the control lamb, was positively correlated with mosquito feedings on young lambs (3 days old), but not on older lambs (6 to 8 weeks old). When Cx. pipiens and Aedes taeniorhynchus were simultaneously given the opportunity to feed on 3-week-old lambs, a significantly higher percentage of engorged Cx. pipiens (94/108, 87%) fed on the viremic lamb than did engorged Ae. taeniorhynchus (187/350, 53%). The data generated by this study (e.g., an excess of feedings on the viremic lambs) warrant further study to determine the significance of this observation under field conditions.     

Mechanical transmission of Rift Valley fever virus by hematophagous Diptera

Experimental studies were conducted to determine if hematophagous Diptera were capable of mechanical transmission of Rift Valley fever (RVF) virus to laboratory animals. All species tested (Glossina morsitans, Aedes aegypti, Aedes taeniorhynchus, Culex pipiens, Stomoxys calcitrans, Lutzomyia longipalpis, and Culicoides variipennis) mechanically transmitted the virus to hamsters. Mechanical transmission rates for G. morsitans ranged from 0-100%, with the probability of mechanical transmission positively correlated with initial viremia titer and negatively correlated with the time after virus exposure. Mechanical transmission of RVF virus to lambs was demonstrated with both G. morsitans and Cx. pipiens. These findings demonstrated that mechanical transmission of RVF virus by hematophagous flies may contribute to the natural transmission and dissemination of this virus.     

Potential for mosquito transmission of attenuated strains of Rift Valley fever virus

Studies were conducted to determine if two attenuated strains of Rift Valley fever (RVF) virus could be transmitted by Culex pipiens mosquitoes. Both strains (RVF MP 12 and T1) replicated in and were transmitted by female Cx. pipiens after intrathoracic inoculation. Mosquitoes also became infected with and transmitted the RVF MP12 strain after ingesting virus from a blood-soaked cotton pledget. However, because of the low viremias produced in infected animals, it is unlikely that mosquitoes would become infected by feeding on an animal inoculated with either of these viruses. Although both strains were transmitted by mosquitoes after intrathoracic inoculation, there was no evidence of reversion to a virulent virus.     

An interspecific mosquito model for the mesenteronal infection barrier to western equine encephalomyelitis virus (Culex tarsalis and Culex pipiens)

Culex tarsalis, a putative mosquito vector of western equine encephalomyelitis (WEE) virus, is susceptible to peroral infection by WEE virus. The nonvector mosquito, Culex pipiens, has a very high peroral threshold of infection and is considered to be refractory. By parenteral inoculation, both mosquito species are equally susceptible. Thus, Cx. tarsalis and Cx. pipiens represent an excellent model system to examine the mechanisms for the mesenteronal infection barrier to WEE virus. Diethylaminoethyl (DEAE)-dextran (1.6 mg/ml), when added to a pledget bloodmeal that contained high concentrations of WEE virus, enhanced peroral infection rates of Cx. pipiens. In Cx. tarsalis, a reduction in infection rates was observed when low WEE concentrations of virus were ingested. DEAE-dextran had no apparent effects on the dissemination of WEE after infection of either mosquito species. It is suggested that the peroral enhancement of WEE viral infection observed with Cx. pipiens may be related to random, nonspecific mechanisms of infection, since it requires high titers of ingested WEE virus. Interference with specific binding of WEE virus to cellular receptor sites is suggested to explain the reduction in WEE viral infection rates in Cx. tarsalis. Altering the pH of the ingested infectious bloodmeal did not affect the WEE viral infection rate of Cx. pipiens, within the range of 6.0-8.5. Cx. tarsalis was optimally infected when the infectious bloodmeal was pH 8.0, and the infection rate was significantly reduced when the infectious blood was at either extreme of the pH range tested. This is again interpreted to indicate that different mechanisms control the peroral infection of Cx. tarsalis and Cx. pipiens with WEE virus. The mesenteronal infection barrier to WEE virus in Cx. pipiens is associated with an inability of the virus to adsorb and/or penetrate mesenteronal epithelial when administered perorally. The barrier is not related to an inability of the Cx. pipiens mesenteronal epithelial cells to support viral multiplication since these cells become infected when the virus is administered parenterally.     

Relative susceptibilties of South Texas mosquitoes to infection with West Nile virus

Three species of mosquitoes (Culex pipiens quinquefasciatus, Aedes aegypti, and Ae. albopictus) collected in Texas were compared with respect to their relative susceptibility to infection with West Nile virus (WNV) strain 114. Oral infection and dissemination rates were 73% infected with 86% dissemination for Ae. aegypti, 13% infected with 100% dissemination for Ae. albopictus, and 100% infected and disseminated for Cx. p. quinquefasciatus. The oral infectious dose required to establish a 50% infection rate was also determined. All feral mosquito species were found to be susceptible to oral infection with WNV (114) in a similar range: Ae. aegypti (6.37 log(10) 50% tissue culture infectious doses [TCID(50)]/5 microL), Ae. albopictus (6.17 log(10) TCID(50)/5 microL), and Cx. p. quinquefasciatus (5.33 log(10) TCID(50)/5 microL). These data demonstrate that both Ae. aegypti and Ae. albopictus from Texas are susceptible to infection with WNV at a similar range to Cx. p. quinquefasciatus and may represent a threat of WNV transmission to humans because of host preferences.     

Susceptibility of Ochlerotatus trivittatus (Coq.), Aedes albopictus (Skuse), and Culex pipiens (L.) to West Nile virus infection

The susceptibility of Ochlerotatus trivittatus (Coq.) to West Nile virus (WNV) was assessed by comparing it to the susceptibility of Aedes albopictus (Skuse), a likely bridge vector, and Culex pipiens (L.), a primary WNV amplifying species. The three species were infected with WNV (NY crow-1999) by feeding on 2-3-day-old chickens with serum virus titers ranging from 10(2.5) to 10(9.5) cell culture infective dose (CID) 50s/mL. The lowest infective titer for Oc. trivittatus and Cx. pipiens was 10(4.5) CID50s/mL. Thirteen percent (4/32) and 2% (1/45) of each species became infected postprandially. Infection rates of the two species increased to 43% (6/14) and 15% (6/40) after blood meals with a titer of 10(5.5) CID50s/mL. In contrast no infection was observed in nine Ae. albopictus that fed among three chickens with titers of 10(4.5) CID50s/mL nor in 41 Ae. albopictus that fed among three chickens with titers of 10(5.0) CID50s/mL. The infective dose 50s for Oc. trivittatus, Cx. pipiens and Ae. albopictus were 10(6.0), 10(6.2), and 10(6.6) CID50s/mL, respectively. Collectively these observations suggest that Oc. trivittatus and Cx. pipiens are more susceptible than Ae. albopictus to WNV when they feed on hosts with WNV titers of <10(7.5) CID50s/mL, but nearly as susceptible with blood meal titers of > or =10(7.5) CID50s/mL. Unpublished studies in our laboratory showed that cottontail rabbits fed on by WNV-infected Oc. trivittatus developed viremias as high as 10(5.5) CID50s/mL serum which exceeds 10 (4.2 (3.4-4.6)) CID50s/mL, the predicted ID10+/-95% CI of Oc. trivittatus. Consequently this mosquito, which also feeds on humans and birds has the potential to serve as a bridge vector and as a maintenance vector among mammals.     

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. Only the mouse monoclonal antibody mixture was tested on the Cx. pipiens. Specific staining of tissues in paraffin sections occurred at dilutions as high as 1:5,000. In both cultured Ae. albopictus cells and sections of Cx. pipiens, only the cytoplasm was positive for antigen. A preliminary list of tissues in Cx. pipiens that exhibited specific or nonspecific staining is given.     

Mosquito feeding modulates Th1 and Th2 cytokines in flavivirus susceptible mice: an effect mimicked by injection of sialokinins, but not demonstrated in flavivirus resistant mice

Culex pipiens and Aedes aegypti mosquitoes were fed on C3H/HeJ mice and systemic cytokine production was quantified from stimulated lymphocytes harvested four to ten days after feeding. Mosquito feeding on C3H/HeJ mice significantly down regulated IFN gamma production seven to ten days post feeding by Cx. pipiens and seven days after Ae aegypti feeding. Th2 cytokines, IL-4 and IL-10, were significantly up regulated 4-7 days after Cx. pipiens and Ae. aegypti feeding. The immunosuppressive effect of Cx. pipiens feeding on systemic cytokine production was not evident in congenic flavivirus resistant (C3H/RV) mice, as systemic IFN gamma and IL-2 were significantly up regulated at days 7 and 10, correlating with a significant decrease in IL-4 10 days after feeding by Cx. pipiens mosquitoes. Inoculation of 5-1000 ng of sialokinin-I into C3H/HeJ mice mimicked the effect of Ae. aegypti feeding by down regulating Th1 cytokines and significantly up regulating Th2 cytokines four days post inoculation. Injections of sialokinin-II resulted in only moderate effects on IFN gamma and IL-4 production seven and ten days after injection. Thus natural feeding by two arbovirus vectors had a profound T cell modulatory effect in vivo in virus susceptible animals which was not demonstrated in the flavivirus resistant host. Moreover, sialokinin-I and sialokinin-II mimicked the effect of mosquito feeding by modulating the host T cell response. These results may lend new insight into specific aspects of the role of the mosquito vector in potentiating virus transmission in the mammalian host.     

Potential vectors of Rift Valley fever virus in the Mediterranean region

We evaluated the ability of three mosquito species (Aedes caspius, Aedes detritus, Culex pipiens), collected in southern France and Tunisia, and of different laboratory-established colonies (Aedes aegypti, Aedes albopictus, Aedes vexans, Anopheles gambiae, Culex pipiens, Culex quinquefasciatus) to disseminate two strains of Rift Valley fever virus (RVFV), the virulent ZH548 and the avirulent Clone 13. After feeding on an infectious blood meal at 10(8.5) plaque-forming units/mL, females were maintained at 30 degrees C for 14 days. Surviving females were tested for the presence of virus on head squashes. Disseminated infection rate corresponds to the number of females with disseminated infection among surviving females. Among field-collected mosquitoes, Cx. pipiens was the most susceptible species with disseminated infection rates ranging from 3.9% to 9.1% for French strains and up to 14.7% for Tunisian strains. Among laboratory-established colonies, Ae. aegypti from Tahiti exhibited the highest disseminated infection rates: 90% when infected with ZH548 and 72.6% with Clone 13. The presence of competent Cx. pipiens in southern France and Tunisia indicates the potential for RVFV epizootics to occur if the virus was introduced into countries of the Mediterranean basin.     

The effect of laboratory colonization on the vector-pathogen interactions of Egyptian Culex pipiens and Rift Valley fever virus

Field and laboratory findings implicated Culex pipiens as a vector of Rift Valley fever (RVF) virus during the 1977-1978 epizootics/epidemics in Egypt. This study evaluated changes in infection and transmission rates, and viral titers in F1 through F16 generation Cx. pipiens mosquitoes orally infected with RVF virus. Infection and transmission rates of RVF virus by this species changed significantly during the colonization process. However, the ultimate viral titers of either the transmitting or the infected nontransmitting mosquitoes were not affected by the colonization process. Following ingestion of virus, Cx. pipiens could be separated into three distinct subpopulations, an uninfected group and two types of infected mosquitoes--transmitters and nontransmitters. Transmitters contained significantly more virus (approximately 100-fold) than nontransmitters. These results demonstrated that not every infected female mosquito should be considered a competent vector, even if the species (population) is known to be a primary vector. Transmission was also accomplished by probing mosquitoes which were unsuccessful in obtaining a blood meal. These data document the long-held suspicion that vector competence studies based upon laboratory-colonized specimens may not represent the field situation.     

Experimental transovarial transmission of St. Louis encephalitis virus by Culex and Aedes mosquitoes

Colonized and field-collected female Culex tarsalis, infected with St. Louis encephalitis, (SLE) virus by intrathoracic inoculation or by feeding on a viremic host, transmitted virus to their F1 adult and/or larval progeny when reared at 18(+/- 1) degree C but not when reared at 27(+/- 1) degrees C. The minimal infection rates (MIR) for different populations of Cx. tarsalis ranged from 1:32 to less than 1:250 (mean = 1:121) for larval progeny and from 1:32 to less than 1:1, 989 (mean = 1:1,571) for adult progeny. SLE virus also was transmitted transovarially by colonized and field-collected populations of Culex pipiens (mean MIRs = 1:340 and 1:1,815 for larval and adult progeny, respectively) and by a field population of Culex quinquefasciatus (MIR = 1:500 and less than 1:246 for larval and adult progeny, respectively), but not by colonized strains of Cx. quinquefasciatus and Culex peus. SLE virus was not recovered in tests on 5,522 Cx. tarsalis and 4,798 Cx. quinquefasciatus that were collected as larvae or pupae from field sites in Southern California and reared to adults at 18 degrees C in the laboratory. Transovarial transmission of SLE virus by Aedes epactius was confirmed and extended to a closely related species, Aedes atropalpus. Efforts to demonstrate transovarial transmission of SLE virus by Aedes melanimon, Aedes sierrensis, and Aedes triseriatus were unsuccessful. Aedes dorsalis, Cx. peus, and Toxorhynchites amboinensis were equally sensitive hosts for viral isolation when inoculated with suspensions of larvae transovarially infected with SLE virus.     

Vector potential of selected North American mosquito species for Rift Valley fever virus

Selected North American mosquito species were evaluated as potential vectors of Rift Valley fever virus. Field populations of Aedes canadensis, Ae. cantator, Ae. excrucians, Ae. sollicitans, Ae. taeniorhynchus, Ae. triseriatus, Anopheles bradleyi-crucians, Culex salinarius, Cx. tarsalis, and Cx. territans perorally exposed to 10(6.2)-10(7.2) plaque forming units of Rift Valley fever virus readily became infected. Infection rates ranged from 51% (65/127) for Cx. salinarius to 96% (64/67) for Ae. canadensis. Disseminated infection rates were generally greater at 14 days than at 7 days after the infectious bloodmeal, and, with the exception of An. bradleyi-crucians, they were not significantly different than the pooled rate of 59% for each species tested. Only 5/55 (9%) of the An. bradleyi-crucians developed a disseminated infection. For most of the species, about half of the mosquitoes with a disseminated infection transmitted an infectious dose of virus to hamsters. While all species, with the exception of An. bradleyi-crucians, transmitted virus, Ae. canadensis, Ae. taeniorhynchus, and Cx. tarsalis had the highest vector potential of the species tested. Following inoculation of approximately 10(1.6) plaque forming units of virus, 100% of the mosquitoes of each species became infected. For most species, transmission rates were similar for inoculated individuals and those that developed a disseminated infection following peroral infection. Viral titers of transmitting and nontransmitting-disseminated individuals were similar for all species tested. These data suggest that, if Rift Valley fever virus was introduced into North America, several mosquito species would be capable of transmitting it.     

Experimental vertical transmission of Japanese encephalitis virus by Culex tritaeniorhynchus and other mosquitoes

Vertical transmission of Japanese encephalitis virus to the F1 adult stage was demonstrated in Culex tritaeniorhynchus, Cx. annulus, Cx. quinquefasciatus, and Armigeres subalbatus. Transmission to the F1 larval stage was demonstrated in Cx. pipiens, Aedes vexans, Ae. alcasidi, and A. flavus. In Cx. tritaeniorhynchus, vertical transmission rates (the percentage of parent females transmitting to progeny) varied (12-100%). Filial infection rates (the percentage of progeny infected) for a given mosquito virus combination were markedly affected by the interval of time between parental infection and oviposition, suggesting that vertical infection was not transovarial in nature but occurred at oviposition. Filial infection rates for Cx. tritaeniorhynchus also varied widely by family and, as measured in F1 larvae, rates in excess of 20% were observed in a family. Filial infection rates in Cx. tritaeniorhynchus F1 adults were about 4 times lower than those in larvae. Japanese encephalitis virus was sexually transmitted from male to female Cx. tritaeniorhynchus.     

Vector competence of five common mosquito species in the People's Republic of China for Western equine encephalitis virus

Two strains of the Western equine encephalitis virus (WEEV) were first detected and isolated in China in 2001. The maintenance and transmission cycles of WEEV in China are currently not well understood, and the mosquito vectors involved in these cycles are unknown. To understand the ability of the local mosquitoes in China to transmit WEEV, the vector competence of five mosquito species, namely, Culex pipiens pallens Coquillett, Cx. p. quinquefasciatus Say, Aedes (Stegomyia) albopictus Skuse, Ae. (Stegomyia) aegypti Linnaeus, and C. tritaeniorhynchus Giles, for WEEV were evaluated. Infection rates for Cx. p. pallens, Cx. p. quinquefasciatus, Cx. tritaeniorhynchus, Ae. Albopictus, and Ae. aegypti were 46%, 60%, 80%, 37%, and 25%, respectively. Dissemination rates for the same species were 60%, 61%, 75%, 55%, and 50%, respectively. Transmission rates were 41%, 53%, 57%, and 45% for Cx. p. pallens, Cx. p. quinquefasciatus, Ae. Albopictus, and Ae. Aegypti, respectively. Infection rates were significantly different between species, but the difference between dissemination and transmission rates were nonsignificant. These results suggest that several local mosquito species in China are competent laboratory vectors for WEEV.     

嗜人按蚊对班氏丝虫易感性的实验观察

本文报告嗜人按蚊对班氏丝虫人工感染的易感性,并和中华按蚊、致倦库蚊和东乡伊蚊的易感性进行比较。在用一名微丝蚴密度为190条/20cmm的班氏微丝蚴血症者血液感染的188只嗜人按蚊、202只中华按蚊、280只致倦库蚊和129只东乡伊蚊中,L_3阳性率分别为35.64％、9.41％、30.0％和65.89％;而在用另一名微丝蚴密度为83条/20cmm的班氏微丝蚴血症者血液感染的188只嗜人按蚊、134只中华按蚊、289只致倦库蚊和176只东乡伊蚊中,L_3阳性率分别为18.09％、3.73％、13.84％和39.77％。结论认为,嗜人按蚊对班氏丝虫人工感染的易感性明显高于中华按蚊,明显低于东乡伊蚊,而与致倦库蚊的易感性处在同一水平上。     

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. However, no antigen was found in the ovarian follicular epithelia, oocytes/nurse cells, or female accessory gland. A viral dissemination index (DI) based on several tissues was determined for each orally infected specimen and used to estimate the extent of viral dissemination in the hemocoel. Plots of DI values as a function of time after ingestion of an infectious blood meal indicated that dissemination was sporadic, and that once virus escapes from the midgut, its spread to other tissues is rapid. The most common sequence of tissue/organ infection following escape of virus from the midgut epithelium appears to be as follows: intussuscepted foregut, fat body, salivary glands and thoracic ganglia, epidermis, and ommatidia of the compound eyes. Reduced fecundity and survival identified in other studies of RVF virus-infected Cx. pipiens are probably explainable on the basis of extensive tissue and organ infection resulting in an overall energy drain. Infection of regulatory tissues and organs such as the ganglia of the central nervous system, neurosecretory cells, and the corpora allata may also have detrimental effects on the overall functioning of a mosquito, even in the absence of widespread infection.