Fig 1 - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
Taxonomic classification of viral sequences in three samples. (A) Viral sequences were classified by host type. Percentage of Mycovirus was too low which hardly can be seen from figure. (B) Viral sequences were classified on family level. The family with reads number less than 10 did not show in figure. Different host types and families were denoted with different colors.
Source publication
Mosquitoes as one of the most common but important vectors have the potential to transmit or acquire a lot of viruses through biting, however viral flora in mosquitoes and its impact on mosquito-borne disease transmission has not been well investigated and evaluated. In this study, the metagenomic techniquehas been successfully employed in analyzin...
Contexts in source publication
Context 1
... total of 92,304 trimmed valid reads were generated after removing sequences shorter than 50 nt and the reads average length was 247 nt (Table 2). 39.2% viral sequences, identified by querying the viral database from NCBI running BLASTn and BLASTx at theEvalue cutoff of 0.01, had high similarities to a variety of different viruses (Table 2), with 3.6% of vertebrate virus, 88% of insect virus, 0.8% of plant virus, 1.87% of bacteriophage and 0.03% of mycovirus ( Fig 1A) among them. However, the abundance and diversity of viral sequences were different in three samples, with the largest viral reads number of Sample III and highest viral diversity in Sample I. Insect virus occupied the most in Samples III, but viral reads of Sample I scatteredly distributed in each virus type, which may attribute to the wide distribution of C. tritaeniorhynchus, the main vector of JEV in China. ...
Citations
... The viral component of the microbiome has also been extensively studied by means of metataxonomic approaches that used meta-RNA shotgun sequencing (e.g., Aragão et al. 2023;Fauver et al. 2016;Feng et al. 2022;Hameed et al. 2021;Liu et al. 2023;Ramírez et al. 2020;Sadeghi et al. 2018;Shi et al. 2015Shi et al. , 2017Thongsripong et al. 2021;Wu et al. 2023;. ...
The incidence of numerous vector-borne diseases (VBDs) has recently increased alarmingly due to various widespread factors, including unplanned urbanization, greater human mobility, environmental changes, vector resistance to insecticides, and evolving pathogens. In this context, the World Health Organization (WHO) has repositioned effective and sustainable vector control as a key approach to prevent and eliminate VBDs. It has been shown that the microbiome influences development, nutrition, and pathogen defense in disease-transmitting vectors such as mosquitoes, sandflies, tsetse flies, triatomine bugs, and ticks. Consequently, understanding the endogenous regulation of vector biology can aid in developing effective approaches for vector control. In this respect, a metatranscriptomic approach analyzes all the expressed RNAs in an environmental sample (meta-RNAs) and can thus reveal how the metabolic activities of the microbiome influence vector biology. This review includes an extensive analysis of available literature on microbial and viral studies for some of the major hematophagous disease-transmitting arthropods, with a focus on studies that used next generation sequencing (NGS) approaches. Since a consensus terminology for these “meta-sequencing analyses” has not yet been established, a definition of these terms is presented here to provide the framework for systematically sorting the available information for each of the VBDs analyzed here to single out metatranscriptomic analyses. Finally, key gaps in knowledge were identified for some of these hematophagous disease-transmitting arthropods which will prove very useful for driving future studies.
... Many studies use the Illumina platform to assess mosquito viromes (Table 5). A few exceptions to that are the Ion platform (Thermo Fisher Scientific) (Öhlund et al. 2019;Amoa-Bosompem et al. 2020), the Oxford Nanopore (Ferreira et al. 2020), and the 454 pyrosequencing platform (Roche) which has since been discontinued (Ng et al. 2011;Shi et al. 2015). Due to the high costs of maintaining NGS sequencing equipment, some research groups choose to delegate the final steps of the sequencing protocol--usually library preparation and the sequencing process per se--to specialized companies. ...
Due to their vectorial capacity, mosquitoes (Diptera: Culicidae) receive special attention from health authorities and entomologists. These cosmopolitan insects are responsible for the transmission of many viral diseases, such as dengue and yellow fever, causing huge impacts on human health and justifying the intensification of research focused on mosquito-borne diseases. In this context, the study of the virome of mosquitoes can contribute to anticipate the emergence and/or the reemergence of infectious diseases. The assessment of mosquito viromes also contributes to the surveillance of a wide variety of viruses found in these insects, allowing the early detection of pathogens with public health importance. However, the study of mosquito viromes can be challenging due to the number and complexities of steps involved in this type of research. Therefore, this article aims to describe, in a straightforward and simplified way, the steps necessary for obtention and assessment of mosquito viromes. In brief, this article explores: the capture and preservation of specimens; sampling strategies; treatment of samples before DNA/RNA extraction; extraction methodologies; enrichment and purification processes; sequencing choices; and bioinformatics analysis.
... None of the 42 virus species we identified was a human pathogen; most are likely to be ISVs, some of which may have a commensal role in their insect hosts. This finding is not surprising as many previous studies on mosquitoes known to vector medically important viruses have not identified human pathogens despite detecting several novel viruses [8,[46][47][48]. Two metatranscriptomic investigations previously conducted in WA by our group also found that the identified viruses were associated with the mosquito host or with other microorganisms within the mosquito [11,14]. ...
Mosquitoes harbor a wide diversity of microorganisms, including viruses that are human pathogens, or that are insect specific. We used metatranscriptomics, an unbiased high-throughput molecular approach, to describe the composition of viral and other microbial communities in six medically important mosquito species from across Western Australia: Aedes vigilax, Culex annulirostris, Cx. australicus, Cx. globocoxitus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. We identified 42 viral species, including 13 novel viruses, from 19 families. Culex mosquitoes exhibited a significantly higher diversity of viruses than Aedes mosquitoes, and no virus was shared between the two genera. Comparison of mosquito populations revealed a heterogenous distribution of viruses between geographical regions and between closely related species, suggesting that geography and host species may play a role in shaping virome composition. We also detected bacterial and parasitic microorganisms, among which Wolbachia bacteria were detected in three members of the Cx. pipiens complex, Cx. australicus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. In summary, our unbiased metatranscriptomics approach provides important insights into viral and other microbial diversity in Western Australian mosquitoes that vector medically important viruses.
... As yet a second study from Turkey only reported minute amounts of virus RNA in worker bees by RNA-seq reads (Mayack and Hakanoğlu, 2022). The biology of the virus and its potential pathogenicity and transmission is not yet known, BSRV appears to be a multi-host pathogen having been also detected by metagenomics analyses in mosquitoes and soybean aphids and maize (Feng et al., 2017, Shi et al., 2015and Wamonje et al., 2017. ...
... The first ORF encodes nonstructural proteins, including RdRp, and the second one encodes capsid proteins. BSRV was first isolated from honeybees (Apis mellifera) [51], and later detected in soybean aphids (Aphis glycines) and Culex tritaeniorhynchus mosquitoes in China [16] and Aphis fabae in Kenya [52]. ...
... Viromes of some blood-sucking ectoparasites, such as mosquitoes and ticks, have been relatively well studied [13][14][15][16][18][19][20] due to their importance as vectors for various human and animal pathogens. Viromes of tabanids, the largest group of hematophagous insects [2,3], remain relatively unstudied. ...
... mellifera) [51], and later in the soybean aphid (A. glycines) and C. tritaeniorhynchus mosquitoes in China [16], as well as in Ap. fabae in Kenya [52]. It should be noted, however, that honeybee BSRV had only 87% similarity with both strains isolated from C. tritaeniorhynchus, as well as with a strain isolated from Ap. fabae [52]. ...
Advances in sequencing technologies and bioinformatics have greatly enhanced our knowledge of virus biodiversity. Currently, the viromes of hematophagous invertebrates, such as mosquitoes and ixodid ticks, are being actively studied. Tabanidae (Diptera) are a widespread family, with members mostly known for their persistent hematophagous behavior. They transmit viral, bacterial, and other pathogens, both biologically and mechanically. However, tabanid viromes remain severely understudied. In this study, we used high-throughput sequencing to describe the viromes of several species in the Hybomitra, Tabanus, Chrysops, and Haematopota genera, which were collected in two distant parts of Russia: the Primorye Territory and Ryazan Region. We assembled fourteen full coding genomes of novel viruses, four partial coding genomes, as well as several fragmented viral sequences, which presumably belong to another twelve new viruses. All the discovered viruses were tested for their ability to replicate in mammalian porcine embryo kidney (PEK), tick HAE/CTVM8, and mosquito C6/36 cell lines. In total, 16 viruses were detected in at least one cell culture after three passages (for PEK and C6/36) or 3 weeks of persistence in HAE/CTVM8. However, in the majority of cases, qPCR showed a decline in virus load over time.
... Thousands of new viruses were discovered, mostly in arthropods [10][11][12]. Currently, viromes of well-established vector invertebrates, such as mosquitoes [13][14][15][16][17] and ixodid ticks [18][19][20] are being actively studied, while other blood-sucking invertebrates are receiving much less attention. To our knowledge, no specific work dedicated to description of the tabanid virome exists. ...
... The first one encodes nonstructural proteins, including RdRp, and the second one encodes capsid proteins. BSRV was first isolated from honeybees (Apis mellifera) [51], and later detected in the soybean aphid (Aphis glycines) and Culex tritaeniorhynchus mosquitoes in China [16], and Aphis fabae in Kenya [52]. ...
... Viromes of some blood-sucking ectoparasites, such as mosquitoes and ticks, have been relatively well-studied [13][14][15][16][18][19][20], due to their importance as a vectors for various human and animals pathogens. Viromes of tabanids, the largest group of haematophagous insects [2,3], remain relatively unstudied. ...
Advances in sequencing technologies and bioinformatics have greatly enhanced our knowledge of virus biodiversity. Currently, viromes of haematophagous invertebrates, such as mosquitoes and ixodid ticks, are actively studied. Tabanidae (Diptera) is a widespread family, with members mostly known for their persistent haematophagous behavior. They transmit viral, bacterial and other pathogens, both biologically and mechanically. However, tabanid viromes remain severely understudied. In this study, we used high-throughput sequencing to describe the viromes of several species of Hybomitra, Tabanus, Chrysops, and Haematopota genera, which were collected in Russia. We assembled 14 full coding genomes of the novel viruses, 4 partial coding genomes, as well as several fragmented viral sequences, which presumably belong to another 12 new viruses. All the discovered viruses were tested on their ability to replicate in the mammalian porcine embryo kidney (PEK), tick HAE/CTVM8, and mosquito C6/36 cell lines. In total, 16 viruses were detected in at least one cell culture after three passages (for PEK and C6/36) or 3 weeks of persistence in HAE/CTVM8. However, in the majority of cases, qPCR showed a decline in virus load over time. Nevertheless, four viruses were detected after six passages.
... By combining bioinformatics tools and virus databases, it is possible to recover assembled contigs and homologous viroid sequences. In 2015, a study of the viral community in three samples, Culex tritaeniorhynchus, Anopheles sinensis, and a mixture pool of Armigeres subalbatus and Culex fatigans from Hubei province, demonstrated a high abundance and diversity of viruses (Shi et al., 2015). In 2016, metagenomics sequencing of adult Anopheles mosquitoes from Liberia, Senegal, and Burkina Faso found a number of virus and virus-like sequences from mosquito midgut contents (Fauver et al., 2016). ...
... In the same year, distinct virome profiles were found in mosquitoes from the Caribbean and locations on the US East Coast (Frey et al., 2016). The viral community of Ae. albopictus also been studied (Shi et al., 2015;He et al., 2021;Calle-Toboń et al., 2022). These studies have increased our understanding of virus diversity in mosquito vectors. ...
... We also detected two unclassified known viruses. The prevalence of Wenzhou sobemo-like virus 4 was reported in samples from Culex quinquefasciatus and Culex tritaeniorhynchus (Shi et al., 2015), and it was detected in female, male, and larvae samples in our study, suggesting possible vertical transmission. We also found several contigs related to known viruses with low identity, indicating the potential presence of novel viruses. ...
Introduction
Aedes albopictus is an aggressive invasive mosquito species widely distributed around the world, and it is also a known vector of arboviruses. Virus metagenomics and RNA interference (RNAi) are important in studying the biology and antiviral defense of Ae. albopictus. However, the virome and potential transmission of plant viruses by Ae. albopictus remain understudied.
Methods
Mosquito samples of Ae. albopictus were collected from Guangzhou, China, and small RNA sequencing was performed. Raw data were filtered, and virus-associated contigs were generated using VirusDetect. The small RNA profiles were analyzed, and maximum-likelihood phylogenetic trees were constructed.
Results
The small RNA sequencing of pooled Ae. albopictus revealed the presence of five known viruses, including Wenzhou sobemo-like virus 4, mosquito nodavirus, Aedes flavivirus, Hubei chryso-like virus 1, and Tobacco rattle virus RNA1. Additionally, 21 new viruses that had not been previously reported were identified. The mapping of reads and contig assembly provided insights into the viral diversity and genomic characteristics of these viruses. Field survey confirmed the detection of the identified viruses in Ae. albopictus collected from Guangzhou.
Discussion
The comprehensive analysis of the virus metagenomics of Ae. albopictus in this study sheds light on the diversity and prevalence of viruses in mosquito populations. The presence of known and novel viruses highlights the need for continued surveillance and investigation into their potential impact on public health. The findings also emphasize the importance of understanding the virome and potential transmission of plant viruses by Ae. albopictus.
Conclusion
This study provides valuable insights into the virome of Ae. albopictus and its potential role as a vector for both known and novel viruses. Further research is needed to expand the sample size, explore additional viruses, and investigate the implications for public health.
... With this new approach, it has been possible to elucidate that Ae. aegypti and Ae. albopictus mosquitoes harbor a rich and diverse virome composed mainly of ISVs [39,107,[113][114][115]. Unlike arboviruses, which have dual host tropism (vertebrates and arthropod vectors), ISVs replicate exclusively in insect populations and cannot replicate in vertebrate cells or infect humans [41,116,117]. ...
... [109,115,[119][120][121]. Phylogenetic analyses on the reconstruction of ancestral traits in ISVs indicate that there are variations in the diversity and abundance of viruses between vector species; however, viral families such as Flaviviridae(positive-sense [+] single-stranded RNA [ssRNA]), Bunyaviridae (negative-sense ssRNA [−]), Rhabdoviridae (ssRNA), Reoviridae (double-stranded RNA [dsRNA]), and Togaviridae (+ssRNA) are shared ...
Aedes aegypti and Aedes albopictus are the main vectors of highly pathogenic viruses for humans, such as dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV), which cause febrile, hemorrhagic, and neurological diseases and remain a major threat to global public health. The high ecological plasticity, opportunistic feeding patterns, and versatility in the use of urban and natural breeding sites of these vectors have favored their dispersal and adaptation in tropical, subtropical, and even temperate zones. Due to the lack of available treatments and vaccines, mosquito population control is the most effective way to prevent arboviral diseases. Resident microorganisms play a crucial role in host fitness by preventing or enhancing its vectorial ability to transmit viral pathogens. High-throughput sequencing and metagenomic analyses have advanced our understanding of the composition and functionality of the microbiota of Aedes spp. Interestingly, shotgun metagenomics studies have established that mosquito vectors harbor a highly conserved virome composed of insect-specific viruses (ISV). Although ISVs are not infectious to vertebrates, they can alter different phases of the arboviral cycle, interfering with transmission to the human host. Therefore, this review focuses on the description of Ae. aegypti and Ae. albopictus as vectors susceptible to infection by viral pathogens, highlighting the role of the microbiota-virome in vectorial competence and its potential in control strategies for new emerging and re-emerging arboviruses.
Graphical Abstract
... In addition, another metagenomic sequencing work revealed ten other viruses infecting An. sinensis namely Culex Bunyavirus 1 (Phenuiviridae), Wutai mosquito phasivirus (Phenuiviridae), Wuhan Mosquito Virus 6 (Orthomyxoviridae), Hubei virga-like virus 1 (Tymoviridae), Yongsan picorna-like virus 4 (Iflaviridae), Wuhan Mosquito Virus 9 (Rhabdoviridae), Culex Y virus (Birnaviridae), Hubei virga-like virus 21 (Tymoviridae) Hubei picorna-like virus 58 (Tymoviridae), and Culex mononega-like virus 1 (Virgaviridae) (He et al., 2021). Furthermore, the porcine parvovirus (PPV) belonging to the genus Parvovirus (subfamily Parvovirinae, family Parvoviridae), was also identified in Infection and dissimilation Jöst et al., 2011;Huhtamo et al., 2013 Anopheles mosquitoes including An. sinensis (Shi et al., 2015;Xia et al., 2018;Hameed et al., 2020). These data show a high diversity of viral communities carried by Anopheles mosquitoes, which may have the potential to infect a wide range of hosts. ...
Malaria remains a vector-borne infectious disease that is still a major public health concern worldwide, especially in tropical regions. Malaria is caused by a protozoan parasite of the genus Plasmodium and transmitted through the bite of infected female Anopheles mosquitoes. The control interventions targeting mosquito vectors have achieved significant success during the last two decades and rely mainly on the use of chemical insecticides through the insecticide-treated nets (ITNs) and indoor residual spraying (IRS). Unfortunately, resistance to conventional insecticides currently being used in public health is spreading in the natural mosquito populations, hampering the long-term success of the current vector control strategies. Thus, to achieve the goal of malaria elimination, it appears necessary to improve vector control approaches through the development of novel environment-friendly tools. Mosquito microbiota has by now given rise to the expansion of innovative control tools, such as the use of endosymbionts to target insect vectors, known as “symbiotic control.” In this review, we will present the viral, fungal and bacterial diversity of Anopheles mosquitoes, including the bacteriophages. This review discusses the likely interactions between the vector microbiota and its fitness and resistance to insecticides.
... Characteristics such as high population density and high anthropophilic behavior of Cx. quinquefasciatus in Brazil can increase the chance of collecting specimens that contain blood in your organism. In this way, increase the probability of identifying circulating arboviruses in the vector population (13,58,59). ...
The (re) emergence of arboviruses around the world is a public health concern once severe outbreaks are usually associated with these infections. The co-circulation of Dengue (DENV), Zika (ZIKV) and Chikungunya (CHIKV) viruses in the past few years has caused a unique epidemic situation in Brazil. The northeast region of the country was the most affected by clinical complications from such arboviruses’ infections, including neurological disorders caused by ZIKV. In this particular region, Aedes mosquitoes are the main vectors of DENV, ZIKV and CHIKV, with Culex quinquefasciatus also considered as a potential vector of ZIKV. Therefore, virological surveillance in mosquitoes contributes to understanding the epidemiological profile of these diseases. Here, we report the circulation of DENV, ZIKV and CHIKV in Aedes spp. and Cx. quinquefasciatus female mosquitoes collected in areas with a high arbovirus circulation in humans in the Metropolitan Region of Recife, Pernambuco, Brazil, during the triple-epidemics of 2015-17. All the field-caught mosquitoes were sent to the laboratory for arbovirus screening after RNA extraction and RT-PCR/RT-qPCR. A total of 6,227 females were evaluated and, as a result, DENV, ZIKV and CHIKV were identified in Ae. aegypti , Ae. taeniorhynchus and Cx. quinquefasciatus mosquito pools. In addition, DENV and ZIKV were isolated in C6/36 cells. In conclusion, it is important to highlight that arbovirus surveillance performed in mosquitoes from DENV-ZIKV-CHIKV hotspots areas can serve as an early-warning system to target vector control actions more efficiently in each studied area.
