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ABSTRACT: The genetic population structure of Aedes (Stegomyia) aegypti (L.), the main vector of dengue virus, is being investigated in areas where a novel dengue suppression program is to be implemented. The aim of the program is to release and establish mosquito populations with impaired virus transmission capabilities. To model effects of the release and devise protocols for its implementation, information about the genetic structure of populations at a range of spatial scales is required.
This study investigates a potential release site in the Hua Sam Rong Subdistrict of Plaeng Yao District, Chachoengsao Province, in eastern Thailand which comprises a complex of five villages within a 10 km radius. Aedes aegypti resting indoors was sampled at four different times of year from houses within the five villages. Genetic markers were used to screen the mosquitoes: two Exon Primed Intron Crossing (EPIC) markers and five microsatellite markers. The raw allele size was determined using several statistical software packages to analyze the population structure of the mosquito. Estimates of effective population size for each village were low, but there was no evidence of genetic isolation by geographic distance.
The presence of temporary genetic structure is possibly caused by genetic drift due to large contributions of adults from a few breeding containers. This suggests that the introduction of mosquitoes into an area needs to proceed through multiple releases and targeting of sites where mosquitoes are emerging in large numbers.
PLoS Neglected Tropical Diseases 01/2013; 7(1):e1913. · 4.69 Impact Factor
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ABSTRACT: Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are related with a variety of
reproductive alterations in their host, the best known being cytoplasmic incompatibility (CI). In this study, prophage WO
gp1, includingWolbachia A andWolbachia B densities were determined during mosquito development by using real-time quantitative PCR (RTQ-PCR). An interesting result
was found for the same flux of prophage WO gp1 andWolbachia B densities in larva, pupa, 3-day-old adult, and 21-day-old adult ofAedes albopictus mosquito (Wolbachia superinfection). The results determined thatWolbachia B and prophage WO gp1, developed with the same fluctuation from larval stage through adult stage. However, the same tendency
was not observed withWolbachia A and phageWO gp1. Hence, this bacteriophage gene is more closely associated withWolbachia B strain than inWolbachia A in their relative density variations. Assesment of the density data reported herein is the first to suggest the possible
association of prophage WO gp1 in CI mechanisms in consequence of the same fluctuation during development withWolbachia B gene density inAedes albopictus mosquito.
Annals of Microbiology 04/2012; 58(2):189-193. · 0.69 Impact Factor
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ABSTRACT: One of the great challenges in the ecology of infectious diseases is to understand what drives the emergence of new pathogens including the relationship between viruses and their hosts. In the case of the emergence of SevereAcute Respiratory Syndrome Coronavirus (SARS-CoV), several studies have shown coronavirus diversity in bats as well as the existence of SARS-CoV infection in apparently healthy bats, suggesting that bats may be a crucial host in the genesis of this disease. To elucidate the biogeographic origin of SARS-CoV and investigate the role that bats played in its emergence, we amplified coronavirus sequences from bat species captured throughout Thailand and assessed the phylogenetic relationships to each other and to other published coronavirus sequences. To this end, RdRp sequence of Coronavirinae was targeted by RT-PCR in non-invasive samples from bats collected in Thailand. Two new coronaviruses were detected in two bat species: one Betacoronavirus in Hipposideros larvatus and one Alphacoronavirus in Hipposiderosarmiger. Interestingly, these viruses from South-East Asia are related to those previously detected in Africa (Betacoronavirus-b) or in Europe (Alphacoronavirus & Betacoronavirus-b). These findings illuminate the origin and the evolutionary history of the SARS-CoV group found in bats by pushing forward the hypothesis of a Betacoronavirus spill-over from Hipposideridae to Rhinolophidae and then from Rhinolophidae to civets and Human. All reported Betacoronaviruses-b (SARS-CoV group) of Hipposideridae and Rhinolophidae respectively cluster in two groups despite their broad geographic distribution and the sympatry of their hosts, which is in favor of an ancient and genetically independent evolution of Betacoronavirus-b clusters in these families. Moreover, despite its probable pathogenicity, we found that a Betacoronavirus-b can persistently infect a medium-sized hipposiderid bat colony. These findings illustrate the importance of the host phylogeny and the host/pathogen ecological interactions in the description and the understanding of pathogen emergence. The host's phylogeny, biogeography and behaviour, combined with already described roles of pathogen plasticity and anthropic changes are likely to be co-factors of disease emergence. Elucidating the common ancestor of Hipposideridae and Rhinolophidae is key to understanding the evolutionary history of actual betacoronaviruses and therefore to get an insight of the deep origin of SARS-CoV.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 07/2011; 11(7):1690-702. · 3.22 Impact Factor
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ABSTRACT: We have used real-time quantitative PCR to measure, for the first time, the relative phage WO-B orf7 density and infection incidence in Aedes albopictus mosquitoes from fields in Thailand. Our results showed that the infection incidence of phage WO-B in this mosquito, sampled from geographically different places in Thailand, was 97.9%. Average relative densities of the offspring were different when collected from diverse parts and reared under the same conditions in the laboratory. Our results also revealed that geographical differences within Thailand did not influence the maternal transmission rate of bacteriophage WO-B. In addition, the orf7 loci might not be strictly associated with Wolbachia, because less than 100% of them were maternally inherited. This discovery does not support the hypothesis that bacteriophage WO-B is involved in Aedes albopictus' cytoplasmic incompatibility. Whether this bacteriophage actually is involved in Wolbachia-induced cytoplasmic incompatibility in this mosquito thus needs further investigation, and additional densities of phage WO-B loci should be integrated.
Current Microbiology 10/2010; 62(3):816-20. · 1.82 Impact Factor
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ABSTRACT: Wolbachia are maternally inherited bacteria that cause various reproductive alterations in their arthropod hosts, including cytoplasmic incompatibility. In this study, we compared mating, oviposition, and host-seeking behavior of Wolbachia-infected (Houston [HOU], Gainesville [GNV]) and Houston uninfected (HT1) Aedes albopictus. In mating assays with virgin mosquitoes, mating success of Wolbachia-infected males was significantly higher than uninfected strains. Mating success was highest with HOU males exposed to infected (95%) and uninfected females (100%), and lowest with HT1 males exposed to infected (20%) and uninfected (25%) females. Results suggested that Wolbachia infection may influence the reproductive behavior of this mosquito. There were no clear differences in oviposition responses between strains, with all strains ovipositing significantly more often on hay infusion and larval rearing water than on water controls and least frequently on 4-methylphenol. Strains of Ae. albopictus females were host-seeking a human when given a choice. Responses to a human arm, acetone, CO2, and dichloromethane were generally higher from the Houston strains than from the GNV strain. Responses of HOU and HT1 females differed from GNV with greater responses to the arm and CO2.
Journal of the American Mosquito Control Association 09/2010; 26(3):265-73. · 0.91 Impact Factor
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ABSTRACT: The highly pathogenic avian influenza (HPAI) H5N1 virus remains a worldwide threat to human and animal health, while the mechanisms explaining its epizootic emergence and re-emergence in poultry are largely unknown. Data from Thailand, a country that experienced significant epidemics in poultry and has recorded suspicious cases of HPAI on a daily basis since 2004, are used here to study the process of emergence. A spatial approach is employed to describe all HPAI H5N1 virus epizootics from 2004 to 2008 and to characterize the pattern of emergence: multiple independent introductions of the virus followed by moderate local spread vs. very rare emergencies followed by strong local spread and rare long range diffusion jumps. Sites where epizootics originate (by foreign introduction, local persistence, or long range jump) were selected from those to which the disease subsequently spreads using a filter based on relative date and position. The spatial distribution of these selected foci was statistically analyzed, and to differentiate environmental factors from long range diffusion, we investigate the relationship of these foci with environmental exposure factors and with rearing characteristics.
During each wave of epizootics, the temporal occurrence of cases did not show a temporal interruption of more than a week. All foci were globally clustered; i.e., more than 90% of cases had a previous case within a 10 km range and a 21 day period of time, showing a strong local spread. We were able to estimate 60 km as the maximum distance for the local farm to farm dissemination process. The remaining "emergent" cases have occurred randomly over Thailand and did not show specific location, clusters, or trends. We found that these foci are not statistically related to specific environmental conditions or land cover characteristics, and most of them may be interpreted as long range diffusion jumps due to commercial practices.
We conclude that only a few foci appear to have been at the origin of each HPAI epidemic wave, leading to the practical action that surveillance and control must focus on farm to farm transmission rather than on emergence or wild fauna.
International Journal of Health Geographics 01/2010; 9:3. · 2.62 Impact Factor
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ABSTRACT: Species of the genus Wolbachia are a group of Rickettsia-like, maternally-inherited bacteria (gram negative), which cause various reproductive alterations in their arthropod and nematode hosts including cytoplasmic incompatibility (CI), male-killing, parthenogenesis and feminization. They can be divided into supergroups such as A and B based on phylogenetic analysis of 16S rDNA sequences. In this study, we examined the relative infection densities of Wolbachia strains among life cycle stages in the mosquito, Aedes albopictus in terms of crowding effect and temperature effect. A. albopictus is known to be superinfected with both A- and B-supergroup Wolbachia which cause CI. The relative Wolbachia densities within each individual mosquito were determined and quantified by using real-time quantitative PCR assay based on the wsp gene. We found that B-supergroup Wolbachia strain densities in this host species were consistently and significantly higher than in the A-supergroup. Larval crowding also reduced adult size of mosquitoes. Our results show clearly that the higher densities of mosquito larvae cause lower densities of Wolbachia strains. Examination of the effect of temperature on Wolbachia density in each stage of the mosquito clearly revealed a significant decrease in bacterial density following exposure to elevated temperature (37 degrees C) in both males and females.
Journal of Invertebrate Pathology 09/2009; 102(3):220-4. · 2.06 Impact Factor
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ABSTRACT: We have evaluated Photorhabdus insect-related protein (Pir) from Photorhabdus asymbiotica against dengue vectors. PirAB shows larvicidal activity against both Aedes aegypti and Aedes albopictus larvae but did not affect the Mesocyclops thermocyclopoides predator. PirAB expressed the strongest toxicity compared to PirA, PirB, or the mixture of PirA plus PirB. Whether the presence of an enterobacterial repetitive intergenic consensus sequence in PirAB, but not in PirA, PirB, or the mixture of PirA plus PirB, has any impact on biological control efficacy needs further investigation.
Applied and environmental microbiology 06/2009; 75(13):4627-9. · 3.69 Impact Factor
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ABSTRACT: Wolbachia is a genus of maternally inherited, intracellular, alpha-Proteobacteria which are widespread among arthropods and cause various reproductive alterations such as cytoplasmic incompatibility and feminization. Wolbachia strains in arthropods have been phylogenetically divided into two supergroups, A and B. Here, we determined the phylogenetic relationships among Wolbachia strains harbored by insects and isopod crustaceans by using groE operon sequences, which encode highly conserved bacterial heat shock proteins. This study includes the first survey for Wolbachia in isopods from a Southeast Asian country, i.e., Thailand. Two isopod species from different parts of Thailand were found to be infected by Wolbachia. Their groE sequences were also included in the phylogenetic analysis. Our results showed clearly that 19 strains from tropical insects, 11 strains from French isopods, and two strains from Thai isopods were in the B supergroup, and 15 strains from tropical insects were in the A supergroup. This is the first report of phylogenetic analysis of a large data set comprising Wolbachia groE sequences from both insects and isopod crustaceans.
ZOOLOGICAL SCIENCE 03/2009; 26(2):171-7. · 0.95 Impact Factor
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ABSTRACT: Female Aedes albopictus mosquitoes from natural populations of different geographical regions of Thailand were collected and allowed to oviposit to determine relative Wolbachia A and Wolbachia B densities of their offspring (F1) by using real-time quantitative PCR (RTQ-PCR). An important aspect of this work is that all Aedes albopictus mosquitoes were collected from the field. Twenty-seven offspring were from diverse areas of Thailand (Songkhla, Konkaen, Chantaburi, and Kanchanaburi). The range of relative Wolbachia A density in F, mosquitoes was from 0.007 to 1250.78 (bacteria-to-host ratio), whereas relative Wolbachia B densities ranged from 0 to 348.2 (bacteria-to-host ratio). These data are in contrast to those from a previous study that showed a very low amount (less than 0.10) of both relative Wolbachia density types for laboratory strains. The percent transmission of Wolbachia density from mother to each individual offspring cannot be predicted and was not related to the sex of the F1. Obtaining confirmation for variations and unpredictable Wolbachia transmission load raises some concerns about using Wolbachia as a gene-driving system in nature for population replacement if Wolbachia density is involved in cytoplasmic incompatibility in this mosquito.
Journal of Vector Ecology 07/2008; 33(1):173-7. · 0.88 Impact Factor
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ABSTRACT: Four mosquito densovirus strains were assayed for mortality and infectivity against Aedes aegypti larvae from different geographic regions. The viral titers were quantified by real-time PCR using TaqMan technology. Firstinstar larvae were exposed to the same titer of each densovirus strain for 48 hours. All strains of densoviruses exhibited larvicidal activity and caused more than 80% mortality and infectivity in the three mosquito strains. AalDNV-exposed larvae had the highest mortality rate. The mean time to death of AalDNV-exposed larvae was shorter than other DNVs-exposed larvae. We can conclude that different densovirus strains exhibit some variations in their pathogenicity to different populations of Ae. aegypti mosquitoes. A few mosquitoes from Chachoengsao and Bangkok exposed to AeDNV and AThDNV survived to the adult stage to lay eggs and showed 22% to 50% vertical transmission in the F1 generation. Phylogenetic analysis of four densovirus strains indicated that mosquito densoviruses are separated into two distinct clades.
The American journal of tropical medicine and hygiene 06/2008; 78(5):784-90. · 2.59 Impact Factor
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ABSTRACT: Mosquito densonucleosis viruses synthesize two non-structural proteins, NS1 and NS2. While NS1 has been studied relatively well, little is known about NS2. Antiserum was raised against a peptide near the N-terminus of NS2, and used to conduct Western blot analysis and immuno-fluorescence assays. Western blots revealed a prominent band near the expected size (41 kDa). Immuno-fluorescence studies of mosquito cells transfected with AeDNV indicate that NS2 has a wider distribution pattern than does NS1, and the distribution pattern appears to be a function of time post-infection. Nuclear localization of NS2 requires intact C-terminus but does not require additional viral proteins. Mutations ranging from complete NS2 knock-out to a single missense amino acid substitution in NS2 can significantly reduce viral replication and production of viable progeny.
Virology 05/2008; 374(1):128-37. · 3.35 Impact Factor
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ABSTRACT: A serological survey of primary school children from six schools in Chachoengsao Province, Thailand, was performed at the end of the peak of dengue transmission. GIS analysis of sero-positive cases was carried out to determine transmission foci. Vector control implementation was conducted in the foci and also within 100 meters around the foci in the treated areas by community participation in collaboration with the local government. Vector control strategies included source reduction together with the use of screen covers, a combination of Bacillus thuringiensis subsp. israelensis and Mesocyclops thermocyclopoides, and lethal ovitraps. Implementation of vector control strategies in the foci was continued until the end of the rainy season. Vector control effectiveness was monitored using entomological, serological, and clinical parameters. Results showed a significant reduction of dengue vectors as well as a decrease in sero-positive children and clinical cases in treated areas when compared with untreated areas.
The American journal of tropical medicine and hygiene 02/2008; 78(1):70-6. · 2.59 Impact Factor
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ABSTRACT: Blood-engorged mosquitoes were collected at poultry farms during an outbreak of highly pathogenic avian influenza in Central Thailand during October 2005. These mosquitoes tested positive for H5N1 virus by reverse transcription-polymerase chain reaction (RT-PCR). Results were confirmed by limited sequencing of the H5 and N1 segments. Infection and replication of this virus in the C6/36 mosquito cell line was confirmed by quantitative real-time PCR. However, transmission by mosquitoes was not evaluated, and further research is needed. Collecting and testing mosquitoes engorged with the blood of domestic or wild animals could be a valuable tool for veterinary and public health authorities who conduct surveillance for H5N1 virus spread.
Vector Borne and Zoonotic Diseases 02/2008; 8(1):105-9. · 2.44 Impact Factor
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ABSTRACT: Dengue hemorrhagic fever is a major public health problem in several countries around the world. Dengue vector surveillance is an important methodology to determine when and where to take the control action. We used a combination of the Global Positioning System (GPS)/Geographic Information System (GIS) technology and the immature sampling count method to improve dengue vector surveillance. Both complete count and sampling count methods were used simultaneously to collect immature dengue vectors in all houses and all containers in one village in eastern Thailand to determine the efficiency of the sampling count technique. A hand-held GPS unit was used to record the location of surveyed houses. Linear regression indicated a high correlation between total immature populations resulting from the complete count and estimates from sampling count of immature stages. The immature survey data and the GPS coordinates of house location were combined into GIS maps showing distribution of immature density and clustering of immature stages and positive containers in the study area. This approach could be used to improve the efficiency and accuracy of dengue vector surveillance for targeting vector control.
The American journal of tropical medicine and hygiene 12/2007; 77(5):897-902. · 2.59 Impact Factor
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ABSTRACT: Geographic information systems (GIS) and remote sensing have been increasingly used in ecology and epidemiology, providing a spatial approach for animal health issues. Recent development of earth environmental satellites--i.e., their growing number, improving sensor resolutions and capabilities--has offered new opportunities to delineate possible habitats and understand animals and associated parasites in their environment, by identifying the nature and structure of land use, hydrological network, soil hydromorphy, and human settlements. Integrated into GIS, remotely sensed and other geo-referenced data allow both spatial and temporal analyses of animal ecology and health. However, a review of their applications has showed the poor quality of data sources and processing used, revealing limitations between theory and practical implementations. As an example, the assessment of the expected distribution of Bandicoot rats, main agricultural pest and vector of zoonoses in Phrae province (North Thailand), illustrates a rational use of spatial analysis, with the choice of relevant data, scales, and processing. Vegetation indices are computed on a TERRA ASTER image and further classified using elevation data. The biotopes of Bandicota indica and Bandicota savilei are delimited, providing a major source of knowledge for rodent and human health analyses.
Annals of the New York Academy of Sciences 11/2006; 1081:17-29. · 3.15 Impact Factor
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ABSTRACT: A community-based dengue vector control trial was conducted at transmission foci in Plaeng Yao District, Chachoengsao Province, eastern Thailand. Implementation was done by the local community in collaboration with local administration, public health, and school authorities. Our cost-effective approaches combined a source reduction campaign with appropriate vector control technologies applied within the foci (within 100 m around the foci) and also within schools attended by children from the treated areas. Vector management measures by local government included cleanup campaigns before the rainy season followed by a routine garbage pickup during the rainy season. Locally made screen covers for water jars, a combination of local Bacillus thuringiensis subsp. israelensis and Mesocyclops thermocyclopoides (copepod), and locally made lethal ovitraps were appropriate technologies used by the community in this campaign. The success of our intervention was evidenced by the significant reduction of dengue vectors and dengue hemorrhagic fever cases in treated areas compared with untreated areas.
Journal of the American Mosquito Control Association 10/2006; 22(3):538-46. · 0.91 Impact Factor
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ABSTRACT: The endosymbiotic bacteria in the genus Wolbachia are capable of inducing a wide range of reproductive abnormalities in their hosts, including cytoplasmic incompatibility (CI), which could lead to the replacement of uninfected host populations with infected ones. Because of this, Wolbachia have attracted considerable interest as a potential mechanism for spreading disease-blocking transgenes through vector populations. Here we report the establishment of double Wolbachia transinfection by direct adult microinjection of Wolbachia from naturally double-infected Aedes albopictus to Aedes aegypti, the most important mosquito vector of infectious viral diseases, and a mosquito in which natural Wolbachia infections are not known to occur. We further demonstrate that incomplete CI is induced in these double-transinfected mosquitoes. Comparisons of fitness traits between naturally uninfected and transinfected Ae. aegypti lines indicated one significant difference in favor of the latter, namely, an increased number of eggs laid. Levels of CI expression corresponded to the Wolbachia density. There were large differences in relative Wolbachia density between reproductive and nonreproductive tissues in both Ae. albopictus and transinfected Ae. aegypti, except Malpighian tubule, which implied the preferred establishment of Wolbachia within reproductive tissue. Results from a simulation model confirm that population replacement by transinfected Ae. aegypti is possible over time. The establishment of Wolbachia double infections in Ae. aegypti by direct adult microinjection and the demonstration of CI expression in this new host suggest that Wolbachia could be experimentally transferred into vector species and could also be used as a gene-driving system to genetically manipulate vector populations.
Proceedings of the National Academy of Sciences 09/2006; 103(33):12534-9. · 9.68 Impact Factor
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ABSTRACT: Severe epidemics of leptospirosis yearly occurred in Thailand since 1997, causing over 1,200 deaths, especially among farmers highly exposed while working in flooded rice fields. Main vectors are wild rodents, spreading the human pathogenic bacteria (spirocheta) in the environment through their urine. Phrae province, which has recorded among the highest incidences, was chosen for a large scale spatial analysis, in collaboration with Phrae provincial Public Health Office. A health geographic database (GIS) was built, using SavGIS, a Geographic Information System (GIS) and Remote Sensing (RS) freeware developed by the Development Research French Institute (IRD), to understand the disease dynamics and assess the villagers’ exposure. GIS and Remote Sensing processing have shown a great potential in analyzing vector-borne diseases but have never been applied to leptospirosis. The land use was extracted and vegetation indices computed from a TERRA ASTER satellite image and described around villages. In a second step, this information was compared with the reported disease incidence. Problems encountered due to the accuracy of biomedical and location-based data are detailed. The results are exposed and discussed in a preventive planning perspective.
Int. J of Geoinformatics. 03/2006; 2(4):43-50.
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ABSTRACT: Research on cultivated shrimp suggests that they have the capability to tolerate viral pathogens in a highly specific manner by mechanisms currently unknown. The phenomenon is difficult to study in detail because they have a generation time of 1-2yr and lack continuous cell lines. Thus, we developed a mosquito-densovirus model to examine whether similar phenomena occur in insects. Serial challenge of five generations with a stock densovirus (AThDNV) resulted in progressive survival increases from 15% to 58%. Prevalence of AThDNV infection in surviving mosquito larvae (confirmed by PCR, histology, in situ hybridization and transmission electron microscopy) was relatively high (e.g. 36% in F2) but they grew normally to establish each succeeding generation. At the end of five generations, comparison of deduced amino acid sequences from genome fragments revealed a significantly higher (p=0.02) estimated prevalence of defective targets in the survivor virus population (29.7%+/-10.0 SD) than in the stored viral population (3.3%+/-5.8 SD). The results paralleled those reported for serially passaged C6/36 mosquito cell cultures infected with a densovirus. There, reduced infection rates are ascribed to the production of defective interfering particles (DIP). Thus, it is possible that the presence of prior AThDNV infections with a high level of DIP contributed to improved survival in our challenged F4 mosquito population. If so, it suggests that persistent viral infections in arthropods may serve in a specific, adaptive manner to reduce the incidence and severity of disease.
Developmental & Comparative Immunology 02/2006; 30(10):878-92. · 3.27 Impact Factor
