Sylvatic Dengue Virus Research Articles

Sylvatic Dengue Viruses Share the Pathogenic Potential of Urban/Endemic Dengue Viruses

Dengue virus (DENV) exists in both sylvatic and urban/endemic ecotypes ([15][1]), and the potential for emergence of sylvatic strains has become a focus of research. Recently Mota and Rico-Hesse ([10][2]) attempted to evaluate the pathogenic potential of viruses belonging to different genetic

Dengue Virus Serotype 2 from a Sylvatic Lineage Isolated from a Patient with Dengue Hemorrhagic Fever

Dengue viruses circulate in both human and sylvatic cycles. Although dengue viruses (DENV) infecting humans can cause major epidemics and severe disease, relatively little is known about the epidemiology and etiology of sylvatic dengue viruses. A 20-year-old male developed dengue hemorrhagic fever (DHF) with thrombocytopenia (12,000/ul) and a raised hematocrit (29.5% above baseline) in January 2008 in Malaysia. Dengue virus serotype 2 was isolated from his blood on day 4 of fever. A phylogenetic analysis of the complete genome sequence revealed that this virus was a member of a sylvatic lineage of DENV-2 and most closely related to a virus isolated from a sentinel monkey in Malaysia in 1970. This is the first identification of a sylvatic DENV circulating in Asia since 1975.

Characterization of a Sylvatic DENV2 Isolated from a DHF Case

Background: Dengue viruses are thought to bemaintained in a sylvatic cycle with the endemic or epidemic viruses arising from ancentral sylvatic strains. In general it is thought that sylvatic dengue viruses are not particularly virulent for humans and may only cause mild disease. In January 2008 we isolated a sylvatic dengue 2 virus from a patient presenting to a fever surveillance clinic. This patient subsequently developed DHF and we present here a description of the case, and early attempts at characterizing the infecting sylvatic virus. Methods: Virus isolation was performed by inoculating C6/36 mosquito cells with 5 ul of serum and incubation of cells at 30 ◦C. One blind passage was performed and when cytopathic effect was observed the isolate was subjected to a Taqman PCR to determine serotype. The nucleotide sequence of the complete E gene was determined by sequencing RT PCR product directly using Big Dye DNA sequencing kit and an ABI 377 sequencer. Dengue virus specific IgM was determined using an IgM Capture ELISA (MAC ELISA) with paired serum specimens tested at 1:100 dilution and dengue IgG was determined using an IgG Capture ELISA (GAC ELISA) in order to determine if the patient had a primary or secondary infection. PRNT50 assays against the sylvatic and currently circulating DENV2 strains were done using serum dilutions in Vero cells in 24 well plates. Western blots were carried out using lysates of infected cells probed with paired sera Results: The phylogenetic analysis of the DENV2 isolated shows that the virus is most closely related to a sylvatic virus P8—1407 isolated in 1970 from a sentinel monkey in Malaysia. This paper will present detailed IgM, IgG, PRNT50 and western blot results for the paired serum against the virus isolated as well as currently circulating DENV2 viruses.

Density-Dependent Competitive Suppression of Sylvatic Dengue Virus by Endemic Dengue Virus in Cultured Mosquito Cells

Mosquito-borne dengue viruses are maintained in two discrete transmission cycles: a sylvatic cycle between nonhuman primates and sylvatic Aedes mosquitoes, and an endemic cycle between humans and peridomestic Aedes (primarily Ae. aegypti and Ae. albopictus). Most sylvatic strains are genetically distinct from endemic strains, and human infections with sylvatic strains have been detected only rarely. Interestingly, sylvatic strains replicate as well as endemic strains in Ae. aegypti and experimental models of replication in humans, suggesting that adaptive constraints may not explain the limited spillover of sylvatic strains into the endemic cycle. Within-host competition is another mechanism known to decrease emergence of strains into occupied niches. In the current study, we examined the magnitude of competitive suppression between sylvatic and endemic dengue strains of different serotypes in pair-wise mixed infections of cultured Ae. albopictus cells to test whether the ecotype or the initial ratio of the two strains influenced the outcome of competition. Strains isolated from nonhuman primates were competitively inferior to those isolated from humans. Moreover, competition was density-dependent; the magnitude of suppression increased as the starting density of a strain relative to its competitor decreased. These data suggest that competitive inferiority in endemic vectors coupled with a numerical disadvantage relative to resident endemic strains could restrict reemergence of sylvatic strains into the endemic cycle and contribute to the ecologically correlated genetic divergence between sylvatic and endemic strains.

Antigenic Relationships between Sylvatic and Endemic Dengue Viruses

Sylvatic dengue viruses (DENVs) are transmitted between non-human primates and arboreal Aedes spp. mosquitoes in Southeast Asia and west Africa. Recent evidence suggests that the risk for re-emergence of sylvatic DENV into the urban endemic/epidemic cycle may be high, which could limit the potential for eradicating the human transmission cycle with vaccines now under development. We assessed the likelihood of sylvatic DENV re-emergence in the face of immunity to current endemic strains or vaccines by evaluating the neutralization capacity of sera from DENV vaccinees and convalescent patients after primary infection with DENV-2 and DENV-3 serotypes. Our data indicate robust homotypic cross-immunity between human sera and sylvatic DENV strains, but limited heterotypic neutralization. Should a licensed vaccine lead to the eradication of the urban transmission cycle in the future, re-emergence of sylvatic strains into the urban cycle would be limited by homotypic immunity mediated by virus-neutralizing antibodies.

Genetic and phenotypic characterization of sylvatic dengue virus type 2 strains

The four serotypes of endemic dengue viruses (DENV) circulate between humans and peridomestic Aedes mosquitoes. At present endemic DENV infect 100 million people per year, and a third of the global population is at risk. In contrast, sylvatic DENV strains are maintained in a transmission cycle between nonhuman primates and sylvatic Aedes species, and are evolutionarily and ecologically distinct from endemic DENV strains. Phylogenetic analyses place sylvatic strains basal to each of the endemic serotypes, supporting the hypothesis that each of the endemic DENV serotypes emerged independently from sylvatic ancestors. We utilized complete genome analyses of both sylvatic and endemic DENV serotype 2 (DENV-2) to expand our understanding of their genetic relationships. A high degree of conservation was observed in both the 5′- and 3′-untranslated genome regions, whereas considerable differences at the nucleotide and amino acid levels were observed within the open reading frame. Additionally, replication of the two genotypes was compared in cultured cells, where endemic DENV strains produced a significantly higher output of progeny in human liver cells, but not in monkey kidney or mosquito cells. Understanding the genetic relationships and phenotypic differences between endemic and sylvatic DENV genotypes may provide valuable insight into DENV emergence and guide monitoring of future outbreaks.

Vector competence of Aedes aegypti populations from Senegal for sylvatic and epidemic dengue 2 virus isolated in West Africa

Vector competence of Aedes aegypti populations from six locations in different bioclimatic zones and habitats of Senegal was assessed for sylvatic (ArD 140875) and epidemic (ArA 6894) dengue 2 virus (DENV-2) isolates. F1 generation mosquitoes were fed with a blood meal containing 10(6.5) and 1.6 x 10(7) tissue culture infectious dose (TCID50) of the sylvatic DENV-2 isolate or 10(6.5) TCID50 of the epidemic isolate and incubated for 14 days. After this period, legs and wings of each individual were pooled and the corresponding body was collected separately. An immunofluorescence assay was used to test bodies and wings/legs for DENV-2 infection separately. The results showed that: (i) Senegalese Ae. aegypti populations have lower vector competence (infection rate 0-26%, dissemination rate 10-100%) for sylvatic and epidemic dengue viruses than populations from Asia and America; (ii) there was no obvious geographic variation in susceptibility among mosquito populations; (iii) the only apparent factor affecting the susceptibility was the virus titre; and (iv) except for that of Kédougou, all populations were able to disseminate the virus. Overall our findings, while extended to four other populations, confirm previous studies showing low susceptibility of Senegalese Ae. aegypti populations.

Sylvatic Dengue Virus Type 2 Activity in Humans, Nigeria, 1966

Using phylogenetic analysis of complete virus genomes from human isolates obtained in Nigeria in 1966, we identified sylvatic dengue virus (DENV) strains from 3 febrile patients. This finding extends current understanding of the role of sylvatic DENV in febrile disease and documents another focus of sylvatic DENV transmission in West Africa.

Dengue Emergence and Adaptation to Peridomestic Mosquitoes

Phylogenetic evidence suggests that endemic and epidemic dengue viruses (DENV), transmitted among humans by the anthropophilic mosquitoes Aedes aegypti and Ae. albopictus, emerged when ancestral, sylvatic DENV transmitted among nonhuman primates by sylvatic Aedes mosquitoes adapted to these peridomestic vectors. We tested this hypothesis by retrospectively examining evidence for adaptation of epidemic and endemic versus sylvatic strains of DENV-2 to Ae. albopictus and Ae. aegypti. First and second-generation offspring of mosquitoes from different geographic regions in the Americas and Southeast Asia were tested for their susceptibility to epidemic/endemic and sylvatic DENV-2 isolates from West Africa, Southeast Asia, and Oceania. Both Aedes species were highly susceptible (up to 100% infected) to endemic/epidemic DENV-2 strains after ingesting artificial blood meals but significantly less susceptible (as low as 0%) to sylvatic DENV-2 strains. Our findings support the hypothesis that adaptation to peridomestic mosquito vectors mediated dengue emergence from sylvatic progenitor viruses.

Evolutionary relationships of endemic/epidemic and sylvatic dengue viruses.

Endemic/epidemic dengue viruses (DEN) that are transmitted among humans by the mosquito vectors Aedes aegypti and Aedes albopictus are hypothesized to have evolved from sylvatic DEN strains that are transmitted among nonhuman primates in West Africa and Malaysia by other Aedes mosquitoes. We tested this hypothesis with phylogenetic studies using envelope protein gene sequences of both endemic/epidemic and sylvatic strains. The basal position of sylvatic lineages of DEN-1, -2, and -4 suggested that the endemic/epidemic lineages of these three DEN serotypes evolved independently from sylvatic progenitors. Time estimates for evolution of the endemic/epidemic forms ranged from 100 to 1,500 years ago, and the evolution of endemic/epidemic forms represents relatively recent events in the history of DEN evolution. Analysis of envelope protein amino acid changes predicted to have accompanied endemic/epidemic emergence suggested a role for domain III in adaptation to new mosquito and/or human hosts.