scholarly journals Australia Ae. aegypti mosquitoes are susceptible to a highly divergent and sylvatic dengue virus type 2 strain infection but are unlikely to transmit

2019 ◽  
Author(s):  
Paul Pickering ◽  
Leon Hugo ◽  
Gregor J Devine ◽  
John G Aaskov ◽  
Wenjun Liu
Keyword(s):  
Cell Culture ◽  
Dengue Virus ◽  
Dengue Fever ◽  
Vector Competence ◽  
Immunosorbant Assay ◽  
Infectious Dose ◽  
Fever Patient ◽  
A Cell ◽  
Divergent Virus

Abstract Background: Humans are the primary hosts of the dengue virus; However, sylvatic cycles of transmission can occur among non-human primates and human encroachment to forested regions can be a source of emergence of new strains. We reported the isolation of a highly divergent and sylvatic DENV-2 strain (QML22) from a dengue fever patient returning Australia from Borneo. The objective of the present study was to evaluate the vector competence of Australian Ae. aegypti mosquitoes for this virus. Methods: Four-day old mosquitoes from two strains of Ae. aegypti from Queensland, Australia, were feed sheep blood meal containing 108 50% cell culture infectious dose per ml (CCID50/ml) of either QML22 or an Australian epidemic DENV serotype 2 strain (QML16) isolated from a dengue fever patient in 2015. Mosquitoes were maintained at 28°C, 75% relative humidity and sampled at 7, 10 and 14 days post-infection (DPI). Live virions in mosquito bodies (abdomen/thorax), legs and wings and saliva expectorates from individual mosquitoes were quantified using a Cell Culture Enzyme-linked Immunosorbant Assay (CCELISA) to determine infection, dissemination and transmission rates. Findings: The infection and dissemination rates of the sylvatic DENV2 strain, QML22, within mosquitoes were significantly lower than that for QML16. While the titres of virus in the bodies of mosquitoes infected with either of these viruses were similar, titres in legs and wings were significantly lower in mosquitoes infected with QML22 at most time points although they reached similar levels by 14 DPI. QML16 was detected in 16% (n = 25) and 28% (n = 25) of saliva expectorates at 10 and 14 DPI, respectively. In contrast, no virus was detected in the saliva expectorates of QML22 infected mosquitoes. Conclusions: Australia urban/peri-urban Ae.aegypti species are susceptible to infection by the sylvatic and highly divergent DENV-2 virus QML22. However, our results indicate that replication of QML22 is attenuated relative to the contemporary strain QML16 and/or a salivary gland infection or escape barrier acts to prevent infection of saliva, potentially preventing onward transmission of this highly divergent virus in Australia.

scholarly journals Australian Aedes aegypti mosquitoes are susceptible to infection with a highly divergent and sylvatic strain of dengue virus type 2 but are unlikely to transmit it

2020 ◽  
Author(s):  
Paul Pickering ◽  
Leon Hugo ◽  
Gregor J Devine ◽  
John G Aaskov ◽  
wenjun Liu
Keyword(s):  
Cell Culture ◽  
Aedes Aegypti ◽  
Dengue Fever ◽  
Vector Competence ◽  
Immunosorbant Assay ◽  
Infectious Dose ◽  
Fever Patient ◽  
A Cell ◽  
Divergent Virus

Abstract Background: Humans are the primary hosts of dengue viruses (DENV). However, sylvatic cycles of transmission can occur among non-human primates and human encroachment into forested regions can be a source of emergence of new strains such as the highly divergent and sylvatic strain of DENV2, QML22, recovered from a dengue fever patient returning to Australia from Borneo. The objective of the present study was to evaluate the vector competence of Australian Aedes aegypti (Ae. aegypti) mosquitoes for this virus. Methods: Four to five day old mosquitoes from two strains of Ae. aegypti from Queensland, Australia, were feed a meal of sheep blood containing 108 50% cell culture infectious dose per ml (CCID50/ml) of either QML22 or an epidemic strain of DENV serotype 2 (QML16) isolated from a dengue fever patient in Australia in 2015. Mosquitoes were maintained at 28°C, 75% relative humidity and sampled 7, 10 and 14 days post-infection (DPI). Live virions in mosquito bodies (abdomen/thorax), legs and wings and saliva expectorates from individual mosquitoes were quantified using a Cell Culture Enzyme-linked Immunosorbant Assay (CCELISA) to determine infection, dissemination and transmission rates. Findings: The infection and dissemination rates of the sylvatic DENV2 strain, QML22, were significantly lower than that for QML16. While the titres of virus in the bodies of mosquitoes infected with either of these viruses were similar, titres in legs and wings were significantly lower in mosquitoes infected with QML22 at most time points although they reached similar levels by 14 DPI. QML16 was detected in 16% (n = 25) and 28% (n = 25) of saliva expectorates at 10 and 14 DPI, respectively. In contrast, no virus was detected in the saliva expectorates of QML22 infected mosquitoes. Conclusions: Australia urban/peri-urban Ae. aegypti species are susceptible to infection by the sylvatic and highly divergent DENV 2 QML22 but replication of QML22 is attenuated relative to the contemporary strain, QML16. A salivary gland infection or escape barrier may be acting to prevent infection of saliva and would prevent onward transmission of this highly divergent virus in Australia.

scholarly journals Australian Aedes aegypti mosquitoes are susceptible to infection with a highly divergent and sylvatic strain of dengue virus type 2 but are unlikely to transmit it

2020 ◽  
Author(s):  
Paul Pickering ◽  
Leon Hugo ◽  
Gregor J Devine ◽  
John G Aaskov ◽  
wenjun Liu
Keyword(s):  
Cell Culture ◽  
Aedes Aegypti ◽  
Dengue Fever ◽  
Vector Competence ◽  
Immunosorbant Assay ◽  
Infectious Dose ◽  
Fever Patient ◽  
A Cell ◽  
Divergent Virus

Abstract Background: Humans are the primary hosts of dengue viruses (DENV). However, sylvatic cycles of transmission can occur among non-human primates and human encroachment into forested regions can be a source of emergence of new strains such as the highly divergent and sylvatic strain of DENV2, QML22, recovered from a dengue fever patient returning to Australia from Borneo. The objective of the present study was to evaluate the vector competence of Australian Aedes aegypti (A. aegypti) mosquitoes for this virus. Methods: Four day old mosquitoes from two strains of A. aegypti from Queensland, Australia, were feed a meal of sheep blood containing 10 8 50% cell culture infectious dose per ml (CCID 50 /ml) of either QML22 or an epidemic strain of DENV serotype 2 (QML16) isolated from a dengue fever patient in Australia in 2015. Mosquitoes were maintained at 28°C, 75% relative humidity and sampled at 7, 10 and 14 days post-infection (DPI). Live virions in mosquito bodies (abdomen/thorax), legs and wings and saliva expectorates from individual mosquitoes were quantified using a Cell Culture Enzyme-linked Immunosorbant Assay (CCELISA) to determine infection, dissemination and transmission rates. Findings: The infection and dissemination rates of the sylvatic DENV2 strain, QML22, were significantly lower than that for QML16. While the titres of virus in the bodies of mosquitoes infected with either of these viruses were similar, titres in legs and wings were significantly lower in mosquitoes infected with QML22 at most time points although they reached similar levels by 14 DPI. QML16 was detected in 16% (n = 25) and 28% (n = 25) of saliva expectorates at 10 and 14 DPI, respectively. In contrast, no virus was detected in the saliva expectorates of QML22 infected mosquitoes. Conclusions: Australia urban/peri-urban A. aegypti species are susceptible to infection by the sylvatic and highly divergent DENV2 virus QML22. Our results indicate that replication of QML22 is attenuated relative to the contemporary strain QML16. Alternatively a salivary gland infection or escape barrier acts to prevent infection of saliva, potentially preventing onward transmission of this highly divergent virus in Australia.

scholarly journals Australian Aedes aegypti mosquitoes are susceptible to infection with a highly divergent and sylvatic strain of dengue virus type 2 but are unlikely to transmit it

2020 ◽  
Author(s):  
Paul Pickering ◽  
Leon Hugo ◽  
Gregor J Devine ◽  
John G Aaskov ◽  
wenjun Liu
Keyword(s):  
Cell Culture ◽  
Aedes Aegypti ◽  
Dengue Fever ◽  
Vector Competence ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infectious Dose ◽  
Fever Patient ◽  
A Cell ◽  
Divergent Virus

Abstract Background: Humans are the primary hosts of dengue viruses (DENV). However, sylvatic cycles of transmission can occur among non-human primates and human encroachment into forested regions can be a source of emergence of new strains such as the highly divergent and sylvatic strain of DENV2, QML22, recovered from a dengue fever patient returning to Australia from Borneo. The objective of the present study was to evaluate the vector competence of Australian Aedes aegypti mosquitoes for this virus. Methods: Four- to five-day-old mosquitoes from two strains of Ae. aegypti from Queensland, Australia, were fed a meal of sheep blood containing 108 50% cell culture infectious dose per ml (CCID50/ml) of either QML22 or an epidemic strain of DENV serotype 2 (QML16) isolated from a dengue fever patient in Australia in 2015. Mosquitoes were maintained at 28 °C, 75% relative humidity and sampled 7, 10 and 14 days post-infection (dpi). Live virions in mosquito bodies (abdomen/thorax), legs and wings and saliva expectorates from individual mosquitoes were quantified using a cell culture enzyme-linked immunosorbent assay (CCELISA) to determine infection, dissemination and transmission rates. Results: The infection and dissemination rates of the sylvatic DENV2 strain, QML22, were significantly lower than that for QML16. While the titres of virus in the bodies of mosquitoes infected with either of these viruses were similar, titres in legs and wings were significantly lower in mosquitoes infected with QML22 at most time points although they reached similar levels by 14 dpi. QML16 was detected in 16% (n = 25) and 28% (n = 25) of saliva expectorates at 10 and 14 dpi, respectively. In contrast, no virus was detected in the saliva expectorates of QML22 infected mosquitoes. Conclusions: Australia urban/peri-urban Ae. aegypti species are susceptible to infection by the sylvatic and highly divergent DENV 2 QML22 but replication of QML22 is attenuated relative to the contemporary strain, QML16. A salivary gland infection or escape barrier may be acting to prevent infection of saliva and would prevent onward transmission of this highly divergent virus in Australia.

scholarly journals A small molecule inhibitor of dengue virus type 2 protease inhibits the replication of all four dengue virus serotypes in cell culture

2015 ◽  
Vol 12 (1) ◽  
pp. 16 ◽  
Author(s):  
Rajendra Raut ◽  
Hemalatha Beesetti ◽  
Poornima Tyagi ◽  
Ira Khanna ◽  
Swatantra K Jain ◽  
...  
Keyword(s):  
Cell Culture ◽  
Dengue Virus ◽  
Virus Type ◽  
Small Molecule ◽  
Small Molecule Inhibitor ◽  
Molecule Inhibitor ◽  
Dengue Virus Serotypes ◽  
Dengue Virus Type 2

scholarly journals Complete Coding Sequences of Dengue Virus Type 2 Strains from Febrile Patients Seen in Malindi District Hospital, Kenya, during the 2017 Dengue Fever Outbreak

2018 ◽  
Vol 6 (15) ◽  
pp. e00076-18 ◽  
Author(s):  
Kimita Gathii ◽  
Josphat N. Nyataya ◽  
Beth K. Mutai ◽  
George Awinda ◽  
John N. Waitumbi
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Virus Type ◽  
District Hospital ◽  
Coding Sequences ◽  
Malindi District ◽  
Dengue Virus Type 2

ABSTRACT We report here 10 complete polyprotein-coding sequences of dengue virus type 2 strains isolated from febrile patients who presented at Malindi District Hospital, Kenya, during a recent dengue fever outbreak. Phylogenetically, all the strains belonged to clonal serotype 2 of the Cosmopolitan genotype.

scholarly journals Replication of Dengue Virus Type 2 in Aedes albopictus Cell Culture

1973 ◽  
Vol 12 (2) ◽  
pp. 275-283 ◽  
Author(s):  
Pantipa Sinarachatanant ◽  
Lloyd C. Olson
Keyword(s):  
Cell Culture ◽  
Dengue Virus ◽  
Virus Type ◽  
Aedes Albopictus ◽  
Albopictus Cell ◽  
Dengue Virus Type 2

scholarly journals Infectious clone construction of dengue virus type 2, strain Jamaican 1409, and characterization of a conditional E6 mutation

2006 ◽  
Vol 87 (8) ◽  
pp. 2263-2268 ◽  
Author(s):  
Dennis J. Pierro ◽  
Ma Isabel Salazar ◽  
Barry J. Beaty ◽  
Ken E. Olson
Keyword(s):  
Cell Culture ◽  
Dengue Virus ◽  
Virus Type ◽  
Mammalian Cells ◽  
Infectious Clone ◽  
Cell Types ◽  
Dengue Virus Type 2

A full-length infectious cDNA clone (ic) was constructed from the genome of the dengue virus type 2 (DENV-2) Jamaica83 1409 strain, pBAC1409ic, by using a bacterial artifical chromosome plasmid system. Infectious virus was generated and characterized for growth in cell culture and for infection in Aedes aegypti mosquitoes. During construction, an isoleucine to methionine (Ile→Met) change was found at position 6 in the envelope glycoprotein sequence between low- and high-passage DENV-2 1409 strains. In vitro-transcribed genomic RNA of 1409ic with E6-Ile produced infectious virions following electroporation in mosquito cells, but not mammalian cells, while 1409ic RNA with an E6-Met mutation produced virus in both cell types. Moreover, DENV-2 1409 with the E6-Ile residue produced syncytia in C6/36 cell culture, whereas viruses with E6-Met did not. However, in vitro cell culture-derived growth-curve data and in vivo mosquito-infection rates revealed that none of the analysed DENV-2 strains differed from each other.

scholarly journals The 1993 dengue 2 epidemic in Charters Towers, North Queensland: clinical features and public health impact

1998 ◽  
Vol 121 (1) ◽  
pp. 151-156 ◽  
Author(s):  
W. J. H. McBRIDE ◽  
H. MULLNER ◽  
J. T. LaBROOY ◽  
I. WRONSKI
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Dengue Infection ◽  
Neutralizing Antibody ◽  
Symptomatic Patient ◽  
Health Impact ◽  
Subclinical Infection ◽  
Dengue Virus Infection ◽  
Dengue Virus Type 2

In 1993 an epidemic caused by dengue virus type 2 occurred in several North Queensland population centres. Charters Towers, estimated population 10000, had 155 officially notified cases. An analysis of symptoms was undertaken using a random sample of 1000 residents to determine specificity of symptoms, the subclinical infection rate, and to establish the true extent of the epidemic. Retrospective diagnoses of dengue fever were based on the presence of both serum dengue 2 neutralizing antibody and presence of symptoms. An estimated 20% of the population had dengue fever. The rate of subclinical infections in this epidemic was 14·6%. There were no symptoms that were specific for dengue fever. Bleeding occurred more frequently in people who recalled a previous dengue infection during a dengue 1 epidemic 12 years earlier (55·6% vs. 16·8%, P=0·003). Surveillance for future epidemics should be based on serological and virological confirmation of dengue virus infection amongst symptomatic patient.

scholarly journals Phylogeny of Dengue virus type 2 isolated in the Central Highlands, Vietnam

2017 ◽  
Vol 65 (2) ◽  
Author(s):  
Tuan Van Le ◽  
Nguyen Thi Tuyet Van ◽  
Nguyen Hoang Quan ◽  
Pham Tho Duoc
Keyword(s):  
Phylogenetic Analysis ◽  
Dengue Virus ◽  
Dengue Fever ◽  
Virus Type ◽  
Public Health Issue ◽  
E Gene ◽  
Study Results ◽  
Central Highlands ◽  
Dengue Virus Type 2

Dengue fever is perhaps the most important viral re-emergent disease especially in tropical and sub-tropical countries, affecting about 50 million people around the world every year. In the Central Highlands regions of Vietnam, dengue fever still remains as a major public health issue. Although four viral serotypes have been currently identified, dengue virus type 2 (DENV-2) was involved in the most important outbreaks during 2010-2012, especially, 2010 when the fatality rate highly increased. Detection of genotype of DENV-2 provided information on origin, distribution and genotype of the virus. In this study, DEN-2 isolated from dengue patients during the 2010-2012 epidemics was amplified and sequenced with E gene. The consensus sequences were aligned with reference E gene sequences of globally available Genbank. Phylogenetic analysis was performed using Neighbor-joining and Kimura 2-parameter model to construct phylogenetic tree. A total of 15 isolates (seven from 2010; one from 2011 and seven from 2012) were obtained from human serum samples. Phylogenetic analysis revealed that Asian genotype 1 is currently circulating locally in Central Highlands region. Isolates of this genotype were closely related to viruses from Thailand, Laos, and Cambodia. It indicated that these epidemics maybe imported into the Central Highlands region from South-East Asia neighbor countries. The study results would help in planning for prevention and control of dengue virus in Vietnam. Continuous monitoring of DENV genotypes is necessary to confirm the current findings and detect possible genotype shifts within the dengue viruses in the future.

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