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A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Virology".

Deadline for manuscript submissions: closed (31 March 2023) | Viewed by 31519

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Dr. Chang-Kweng Lim

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Laboratory of Arboviruses, Department of Virology 1, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan
Interests: developing animal models; laboratory diagnostics; viral replication and assembly, and viral pathology and molecular epidemiology
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Special Issue Information

Dear Colleagues,

Arbovirus diseases such as dengue fever, zika virus disease, yellow fever virus, tick-borne encephalitis, West Nile encephalitis, and Japanese encephalitis are emerging and re-emerging human illnesses transmitted by arthropod vectors. Up to 390 million people are infected every year with the dengue virus. Millions of people are infected with other arboviruses such as chikungunya virus, Zika virus, and Japanese encephalitis virus. The burden of these diseases is highest in tropical and subtropical areas, and they have afflicted populations and claimed lives in many countries. Furthermore, arboviruses are increasingly prevalent in more temperate regions, resulting in even more people being at risk. The distribution of arbovirus diseases is determined by complex demographic, environmental, social factors, and global travel and trade. This Special Issue of Microorganisms highlights recent advances in molecular biology, molecular evolution, diagnostics, animal models, and the control of emerging arboviruses such as Usutu virus, Ross River virus, Muko virus, and Mayaro virus.

Dr. Chang-Kweng Lim
Guest Editor

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Keywords

dengue virus
zika virus
chikungunya virus
Japanese encephalitis virus
animal model and pathogenesis
diagnostics and prevention

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Published Papers (10 papers)

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Research

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18 pages, 2601 KiB

Open AccessArticle

A Six Years (2010–2016) Longitudinal Survey of the Four Serotypes of Dengue Viruses in Lao PDR

by Charlotte Balière, Elodie Calvez, Jean-Michel Thiberge, Somphavanh Somlor, Mathias Vandenbogaert, Marc Grandadam and Valérie Caro

Microorganisms 2023, 11(2), 243; https://doi.org/10.3390/microorganisms11020243 - 18 Jan 2023

Cited by 1 | Viewed by 2399

Abstract

Dengue fever is the most prevalent arthropod-borne viral infection of humans in tropical and subtropical countries. Since 1979, dengue has been reported to be endemic in the Lao People’s Democratic Republic (PDR), as in many countries in Southeast Asia, with a complex circulation of the four dengue viruses’ serotypes (DENV-1 to DENV-4). By sequencing the complete envelope protein, we explored a panel of samples from five Lao Provinces (Vientiane capital, Luangprabang, Bolikhamxay, Saravane, Attapeu) to enrich knowledge about the co-circulation of DENVs in Lao PDR between 2010 and 2016. Phylogenetic analyses highlighted the specific circulation of DENV-1 genotype I, DENV-2 genotype Asian I, DENV-4 genotype I and the co-circulation of DENV-3 genotype II and III. The continuous co-circulation of the four serotypes was underlined, with genotype or cluster shifts among DENV-3 and DENV-1. These data suggested the emergence or re-emergence of DENV strains associated with epidemic events, potentially linked to the exchanges within the territory and with neighboring countries. Indeed, the increasing local or regional connections favored the dissemination of new isolates or new clusters around the country. Since 2012, the surveillance and alert system created in Vientiane capital by the Institut Pasteur du Laos appears to be a strategic tool for monitoring the circulation of the four serotypes, especially in this endemic country, and allows for improving dengue epidemiological knowledge to anticipate epidemic events better. Full article

(This article belongs to the Special Issue Arboviruses)

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12 pages, 1920 KiB

Open AccessArticle

The Colombian Zika Virus Isolate (COL345Si) Replicates in Prostate Adenocarcinoma Cells and Modulates the Antiviral Response

by Yaneth Miranda Brand, Astrid M. Bedoya, Liliana Betancur-Galvis and Juan Carlos Gallego-Gómez

Microorganisms 2022, 10(12), 2420; https://doi.org/10.3390/microorganisms10122420 - 7 Dec 2022

Viewed by 1721

Abstract

Zika virus (ZIKV), a flavivirus that is mainly transmitted by A. aegypti and A. albopictus and sexual transmission, has been documented and described. The ZIKV RNA detection in the semen of vasectomized men indicates that accessory glands such as the prostate could be a site of virus replication. In this study, we characterized the ZIKV infection, evaluated the antiviral profile, and demonstrated the AXL and TIM-1 expression on the PC3 prostate cell line. It was also determined that PC3 cells are susceptible and permissive to ZIKV infection without altering the cell viability or causing a cytopathic effect. The antiviral profile suggests that the PC3 cells modulate the antiviral response through the suppressor molecule expression, SOCS-1, during a ZIKV infection. Full article

(This article belongs to the Special Issue Arboviruses)

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15 pages, 756 KiB

Open AccessArticle

Serologic Investigation on Tick-Borne Encephalitis Virus, Kemerovo Virus and Tribeč Virus Infections in Wild Birds

by Katarína Peňazziová, Ľuboš Korytár, Ivana Cingeľová Maruščáková, Petra Schusterová, Alexander Loziak, Soňa Pivka, Anna Ondrejková, Juraj Pistl and Tomáš Csank

Microorganisms 2022, 10(12), 2397; https://doi.org/10.3390/microorganisms10122397 - 2 Dec 2022

Cited by 4 | Viewed by 2291

Abstract

The present study reports on serosurvey on the tick-borne encephalitis virus European subtype (TBEV; genus Flavivirus), and the tick-borne Kemerovo (KEMV) and Tribeč (TRBV) orbivirus (genus Orbivirus) infections in tick-infested and non-infested birds. No virus RNA was detected in the blood clots. Birds were infested mostly by Ixodes ricinus, but Haemaphysalis concinna and I. frontalis were observed too. TBEV, KEMV and TRBV neutralising antibodies (NAb) were detected in the screening microtitration neutralisation test (μVNT). Seropositive samples were further examined in simultaneous μVNT to distinguish TBEV infection from WNV and USUV. KEMV and TRBV infections were also further examined by μVNT against each other. The demonstrated results point to increased TBEV and TRBV seroprevalence in birds over the past several years. This is the first study on KEMV infection in the Slovak bird population, and seropositive juvenile birds suggest its occurrence in a new geographic area. The results indicate the significance of tick infestation rates, seropositivity and specific NAb titre. The reservoir role of birds for TBEV, KEMV and TRBV remains unclear. However, targeted monitoring of birds and vectors is an effective measure of surveillance of arbovirus introduction into new geographic areas. Full article

(This article belongs to the Special Issue Arboviruses)

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14 pages, 11773 KiB

Open AccessArticle

A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya

by Leonardo Assis da Silva, Monique da Rocha Queiroz Lima, Brenda Rabello de Camargo, Dyeferson Kened da Silva Coelho Guimarães, Anabele Azevedo Lima Barbastefano, Raquel Curtinhas de Lima, Paulo Vieira Damasco, Rivaldo Venâncio da Cunha, Luiz José de Souza, Elzinandes Leal de Azeredo, Luzia Maria de-Oliveira-Pinto, Tatsuya Nagata, Daniel M. P. Ardisson-Araújo, Flavia Barreto dos Santos and Bergmann Morais Ribeiro

Microorganisms 2022, 10(7), 1451; https://doi.org/10.3390/microorganisms10071451 - 18 Jul 2022

Cited by 3 | Viewed by 2371

Abstract

Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance. Full article

(This article belongs to the Special Issue Arboviruses)

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16 pages, 2970 KiB

Open AccessArticle

Lineage Replacement Associated with Fitness Gain in Mammalian Cells and Aedes aegypti: A Catalyst for Dengue Virus Type 2 Transmission

by Cheong Huat Tan, Hapuarachchige Chanditha Hapuarachchi, Li Kiang Tan, Pei Sze Jeslyn Wong, Mei Zhi Irene Li, Wing Yan Wong and Lee Ching Ng

Microorganisms 2022, 10(6), 1100; https://doi.org/10.3390/microorganisms10061100 - 26 May 2022

Cited by 5 | Viewed by 2254

Abstract

Shifting of virus serotypes and clade replacement events are known to drive dengue epidemics. However, only a few studies have attempted to elucidate the virus attributes that contribute to such epidemics. In 2007, Singapore experienced a dengue outbreak affecting more than 8000 individuals. The outbreak ensued with the shuffling of dominant clades (from clade I to clade II) of Dengue virus 2 (DENV-2) cosmopolitan genotype, at a time when the Aedes premise index was significantly low. Therefore, we hypothesized that clade II had higher epidemic potential and fitness than clade I. To test this hypothesis, we tested the replication and apoptotic qualities of clade I and II isolates in mammalian cells and their ability to infect and disseminate in a field strain of Ae. Aegypti. Our findings indicated that clade II replicated more efficiently in mammalian cells than clade I and possessed higher transmission potential in local vectors. This could collectively improve the epidemic potential of clade II, which dominated during the outbreak in 2007. The findings exemplify complex interactions between the emergence, adaptation and transmission potential of DENV, and testify the epidemiological importance of a deeper understanding of virus and vector dynamics in endemic regions. Full article

(This article belongs to the Special Issue Arboviruses)

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16 pages, 1882 KiB

Open AccessEditor’s ChoiceArticle

Enhanced Immunogenicity of Inactivated Dengue Vaccines by Novel Polysaccharide-Based Adjuvants in Mice

by Shuenn-Jue Wu, Dan Ewing, Appavu K. Sundaram, Hua-Wei Chen, Zhaodong Liang, Ying Cheng, Vihasi Jani, Peifang Sun, Gregory D. Gromowski, Rafael A. De La Barrera, Megan A. Schilling, Nikolai Petrovsky, Kevin R. Porter and Maya Williams

Microorganisms 2022, 10(5), 1034; https://doi.org/10.3390/microorganisms10051034 - 16 May 2022

Cited by 2 | Viewed by 3621

Abstract

Dengue fever, caused by any of four dengue viruses (DENV1-4), is a major global burden. Currently, there is no effective vaccine that prevents infection in dengue naïve populations. We tested the ability of two novel adjuvants (Advax-PEI and Advax-2), using aluminum hydroxide (alum) as control, to enhance the immunogenicity of formalin- or psoralen-inactivated (PIV or PsIV) DENV2 vaccines in mice. Mice were vaccinated on days 0 and 30, and serum samples were collected on days 30, 60, 90, and 101. Neutralizing antibodies were determined by microneutralization (MN) assays, and the geometric mean 50% MN (MN₅₀) titers were calculated. For the PIV groups, after one dose MN₅₀ titers were higher in the novel adjuvant groups compared to the alum control, while MN₅₀ titers were comparable between the adjuvant groups after the second dose. For the PsIV groups, both novel adjuvants induced higher MN₅₀ titers than the alum control after the second dose. Spleen cells were collected on days 45 and 101 for enzyme-linked immunospot (ELISPOT) for IFNγ and IL4. Both PIV and PsIV groups elicited different degrees of IFNγ and IL4 responses. Overall, Advax-2 gave the best responses just ahead of Advax-PEI. Given Advax-2’s extensive human experience in other vaccine applications, it will be pursued for further development. Full article

(This article belongs to the Special Issue Arboviruses)

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21 pages, 3769 KiB

Open AccessArticle

Spread of West Nile Virus and Usutu Virus in the German Bird Population, 2019–2020

by Ute Ziegler, Felicitas Bergmann, Dominik Fischer, Kerstin Müller, Cora M. Holicki, Balal Sadeghi, Michael Sieg, Markus Keller, Rebekka Schwehn, Maximilian Reuschel, Luisa Fischer, Oliver Krone, Monika Rinder, Karolin Schütte, Volker Schmidt, Martin Eiden, Christine Fast, Anne Günther, Anja Globig, Franz J. Conraths, Christoph Staubach, Florian Brandes, Michael Lierz, Rüdiger Korbel, Thomas W. Vahlenkamp and Martin H. Groschup Show full author list Hide full author list

Microorganisms 2022, 10(4), 807; https://doi.org/10.3390/microorganisms10040807 - 12 Apr 2022

Cited by 28 | Viewed by 4751

Abstract

West Nile virus (WNV) and Usutu virus (USUV) are important flaviviruses circulating in Germany. While USUV was first reported more than 10 years ago, WNV has only reached the country in 2018. Wild birds are important amplifying hosts for both viruses. Therefore, we have been monitoring the bird population in different regions of Germany by a previously established network for many years. This report summarizes the results of molecular and/or serological methods of 2345 blood samples from birds of 22 different orders and over 2900 bird carcasses from 2019 and 2020. USUV RNA circulation was found in different regions of Germany, with emphasis on USUV lineages Europe 3 and Africa 3. Increased evidence of USUV lineage Europe 2 was detected in eastern Germany. WNV RNA was found only in birds from the eastern part of the country. The seroprevalence for USUV was between 3.11% and 7.20% in all three regions investigated, whereas the WNV seroprevalence spanned from 14.77% to 16.15% in eastern Germany, with a noticeable tendency for a westward and southward expansion in both years. Thus, wild bird monitoring for WNV and USUV can serve as an early warning system for a human exposure risk. Full article

(This article belongs to the Special Issue Arboviruses)

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14 pages, 1399 KiB

Open AccessArticle

Genotype-Dependent Immunogenicity of Dengue Virus Type 2 Asian I and Asian/American Genotypes in Common Marmoset (Callithrix jacchus): Discrepancy in Neutralizing and Infection-Enhancing Antibody Levels between Genotypes

by Nor Azila Muhammad Azami, Meng Ling Moi, Yasushi Ami, Yuriko Suzaki, Satoshi Taniguchi, Shigeru Tajima, Masayuki Saijo, Tomohiko Takasaki, Ichiro Kurane and Chang-Kweng Lim

Microorganisms 2021, 9(11), 2196; https://doi.org/10.3390/microorganisms9112196 - 21 Oct 2021

Viewed by 2211

Abstract

Owing to genotype-specific neutralizing antibodies, analyzing differences in the immunogenic variation among dengue virus (DENV) genotypes is central to effective vaccine development. Herein, we characterized the viral kinetics and antibody response induced by DENV type 2 Asian I (AI) and Asian/American (AA) genotypes using marmosets (Callithrix jacchus) as models. Two groups of marmosets were inoculated with AI and AA genotypes, and serial plasma samples were collected. Viremia levels were determined using quantitative reverse transcription-PCR, plaque assays, and antigen enzyme-linked immunosorbent assay (ELISA). Anti-DENV immunoglobulin M and G antibodies, neutralizing antibody titer, and antibody-dependent enhancement (ADE) activity were determined using ELISA, plaque reduction neutralization test, and ADE assay, respectively. The AI genotype induced viremia for a longer duration, but the AA genotype induced higher levels of viremia. After four months, the neutralizing antibody titer induced by the AA genotype remained high, but that induced by the AI genotype waned. ADE activity toward Cosmopolitan genotypes was detected in marmosets inoculated with the AI genotype. These findings indicate discrepancies between heterologous genotypes that influence neutralizing antibodies and viremia in marmosets, a critical issue in vaccine development. Full article

(This article belongs to the Special Issue Arboviruses)

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Review

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20 pages, 1330 KiB

Open AccessReview

Latest Advances in Arbovirus Diagnostics

by Jano Varghese, Imesh De Silva and Douglas S. Millar

Microorganisms 2023, 11(5), 1159; https://doi.org/10.3390/microorganisms11051159 - 28 Apr 2023

Cited by 9 | Viewed by 4596

Abstract

Arboviruses are a diverse family of vector-borne pathogens that include members of the Flaviviridae, Togaviridae, Phenuviridae, Peribunyaviridae, Reoviridae, Asfarviridae, Rhabdoviridae, Orthomyxoviridae and Poxviridae families. It is thought that new world arboviruses such as yellow fever virus emerged in the 16th century due to the slave trade from Africa to America. Severe disease-causing viruses in humans include Japanese encephalitis virus (JEV), yellow fever virus (YFV), dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKV), Crimean–Congo hemorrhagic fever virus (CCHFV), severe fever with thrombocytopenia syndrome virus (SFTSV) and Rift Valley fever virus (RVFV). Numerous methods have been developed to detect the presence of these pathogens in clinical samples, including enzyme-linked immunosorbent assays (ELISAs), lateral flow assays (LFAs) and reverse transcriptase–polymerase chain reaction (RT-PCR). Most of these assays are performed in centralized laboratories due to the need for specialized equipment, such as PCR thermal cyclers and dedicated infrastructure. More recently, molecular methods have been developed which can be performed at a constant temperature, termed isothermal amplification, negating the need for expensive thermal cycling equipment. In most cases, isothermal amplification can now be carried out in as little as 5–20 min. These methods can potentially be used as inexpensive point of care (POC) tests and in-field deployable applications, thus decentralizing the molecular diagnosis of arboviral disease. This review focuses on the latest developments in isothermal amplification technology and detection techniques that have been applied to arboviral diagnostics and highlights future applications of these new technologies. Full article

(This article belongs to the Special Issue Arboviruses)

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21 pages, 1054 KiB

Open AccessReview

Arbovirus Epidemiology: The Mystery of Unnoticed Epidemics in Ghana, West Africa

by Eric Agboli, Alexandru Tomazatos, Oumou Maiga-Ascofaré, Jürgen May, Renke Lühken, Jonas Schmidt-Chanasit and Hanna Jöst

Microorganisms 2022, 10(10), 1914; https://doi.org/10.3390/microorganisms10101914 - 27 Sep 2022

Cited by 2 | Viewed by 3693

Abstract

It is evident that all the countries surrounding Ghana have experienced epidemics of key arboviruses of medical importance, such as the recent dengue fever epidemic in Burkina Faso. Therefore, Ghana is considered a ripe zone for epidemics of arboviruses, mainly dengue. Surprisingly, Ghana never experienced the propounded deadly dengue epidemic. Indeed, it is mysterious because the mosquito vectors capable of transmitting the dengue virus, such as Aedes aegypti, were identified in Ghana through entomological investigations. Additionally, cases may be missed, as the diagnostic and surveillance capacities of the country are weak. Therefore, we review the arbovirus situation and outline probable reasons for the epidemic mystery in the country. Most of the recorded cases of arbovirus infections were usually investigated via serology by detecting IgM and IgG immunoglobulins in clinical samples, which is indicative of prior exposure but not an active case. This led to the identification of yellow fever virus and dengue virus as the main circulating arboviruses among the Ghanaian population. However, major yellow fever epidemics were reported for over a decade. It is important to note that the reviewed arboviruses were not frequently detected in the vectors. The data highlight the necessity of strengthening the diagnostics and the need for continuous arbovirus and vector surveillance to provide an early warning system for future arbovirus epidemics. Full article

(This article belongs to the Special Issue Arboviruses)

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