A new cluster of rhabdovirus detected in field-caught sand flies (Diptera: Psychodidae: Phlebotominae) collected from southern Thailand
Parasites & Vectors volume 14, Article number: 569 (2021)
The distribution of phlebotomine sand flies is changing rapidly due to climate change. This issue has implications for the epidemiology of sand fly-borne diseases, especially sand fly-associated viruses. Few studies concerning sand fly-associated viruses have been conducted in Thailand. Therefore, this study aimed to perform a molecular survey of groups of pathogenic RNA viruses belonging to the Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae in sand fly samples collected from southern Thailand.
Sand flies were collected at two locations in Trang and Songkhla provinces of southern Thailand, and individual sand fly samples were processed for species identification and virus detection. The Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae molecular determination was performed by RT-PCR, and positive samples were identified by cloning and sequencing, cell culture inoculation, and phylogenetic analysis.
The results presented in this study were based on the analysis of a total of 331 female sand flies. This molecular study revealed evidence of Rhabdoviridae family virus presence in Phlebotomus papatasi (3/331, 0.9%). The findings demonstrated a new cluster of rhabdovirus that was closely related to Bactrocera dorsalis sigmavirus strain BDSV.abc5 and the lineages of insect-specific Rhabdoviridae. In addition, the Bayesian tree suggested that the common ancestor of this group was the dimarhabdovirus clade. It was assumed that the virus may have switched hosts during its evolution. However, the detection of Orbivirus, Phlebovirus, and Flavivirus genera using specific primers for RT-PCR was negative in the collected sand flies.
There is limited knowledge on the genetic diversity and ecology of Rhabdoviridae in Thailand. This is the first data regarding the circulation of Rhabdoviridae in Ph. papatasi from Thailand. We found a new cluster of rhabdoviruses that was close to the new B. dorsalis sigmavirus. It is possible that there is a great deal of diversity in this family yet to be discovered, and a more extensive survey for new rhabdoviruses may uncover viruses from a wide diversity of host taxa and broaden our understanding of the relationships among the Rhabdoviridae.
Phlebotomine sand flies (Diptera: Psychodidae) are a medically important group of insects widely distributed throughout the world. They are responsible for causing several diseases in humans and other animals, including leishmaniasis, sand fly fever, and human bartonellosis [1,2,3]. In southern Thailand, several sand fly-borne pathogens have been detected in various sand fly species, including Leishmania parasites in Sergentomyia (Neophlebotomus) gammae, Se. khawi, and Se. (Parrotomyia) barraudi sand flies, and Trypanosoma parasites in Phlebotomus stantoni, Se. khawi, Se. indica, Se. anodontis, Ph. asperulus, and Ph. betisi species [4, 5]. Sand fly fever is still a significant health problem in many regions of the world, particularly in the Mediterranean Basin, the Middle East, Africa, Central Asia, and Europe. [6, 7]. The common serotypes of phleboviruses (family Bunyaviridae, genus Phlebovirus) are sand fly fever Sicilian virus (SFSV), sand fly fever Naples virus (SFNV) and Toscana virus (TOSV), which are transmitted to humans by different species of sand flies, especially Phlebotomus species [8, 9]. Moreover, there are other viruses that can be transmitted by sand flies, such as family Rhabdoviridae , family Reoviridae , and family Flaviviridae . Interestingly, rhabdoviruses can infect the cytoplasm of host cells. They possess an approximately 12-kilobase (kb) genome that usually consists of five genes (3′-N-P-M-G-L-5′) . In 2015, 13 genera of rhabdoviruses were available at http://www.ictvonline.org/virusTaxonomy.asp, comprising Cytorhabdovirus, Dichorhavirus, Ephemerovirus, Lyssavirus, Novirhabdovirus, Nucleorhabdovirus, Perhabdovirus, Sigmavirus, Sprivivirus, Tibrovirus, Tupavirus, Varicosavirus, and Vesiculovirus [14, 15]. Similar to the majority of the known dimarhabdoviruses (e.g. genera Sigmavirus, Vesiculovirus, and Ephemerovirus), cytorhabdoviruses, nucleorhabdoviruses, and rhabdoviruses are carried by arthropods . Despite their crucial roles in the epidemiology of the disease, in-depth knowledge of sand fly-associated viruses is still scarce. In addition, several factors associated with climate change, human activity, and different geographical regions affect sand fly-borne pathogen distribution in some parts of the world . The most recent report found that the Orbivirus genus in Idiophlebotomus spp., Phlebotomus papatasi, and Se. khawi from southern Thailand was closely related to Changuinola virus (CGLV) . However, it is the only report of survey studies of the distribution of sand fly-associated viruses in Thailand. Therefore, the objective of this study was to survey and screen for the presence of RNA viruses, including the genera Orbivirus, Phlebovirus, and Flavivirus and family Rhabdoviridae, in field-caught sand flies collected from the southern region of Thailand. The detection method used was reverse transcription polymerase chain reaction (RT-PCR) with the use of viral-specific primers to precisely target each virus and sequence analysis. This technique provides a rapid means for obtaining expansive results that facilitate the investigation of the phylogenetic relationships of the virus and its host. Moreover, identification of the host and virus associations remains a key challenge for virus research. The impacts of virus screening in sand flies from Thailand can offer an important contribution to the establishment of vector control programs for sand fly-associated viruses in the country and the region.
Sand fly collection and species identification
A total of 331 female sand flies were collected in Huai Yot District, Trang Province (7°47′6″N, 99°38′6″E), and in Na Thawi District, Songkhla Province (6°39′58.9″N, 100°42′34.6″E), Thailand, during the period 14–20 December 2020. The sand flies were collected using five to seven miniature Centers for Disease Control and Prevention (CDC) light traps, which were installed and hung from tree branches or hooks at a height of 1 m measuring from the trap hood to the ground. All traps were activated for 12 h from 6.00 p.m. to 6.00 a.m. Insects were collected from the light traps the following day. These sand flies were stored in 2-ml cryovials in liquid nitrogen and then transported to the laboratory for sorting and taxonomic identification. Individual female sand flies were placed on a sterilized slide and dissected under a stereomicroscope equipped with an ice block, using 26G × 1/2″ sterile needles. Head and genitalia were mounted with Hoyer’s medium on a slide, which was later used for morphological species identification based on keys proposed by Lewis , Phumee et al. , and Depaquit et al. . Molecular identification was performed to confirm the sand fly species using mitochondrial cytochrome b gene (CytB)-PCR . The abdomen and thorax of each female specimen were transferred to a sterile 1.5-ml Eppendorf tube containing 200 μl of 1× phosphate-buffered saline (PBS). All samples were then stored at −80 °C until viral screening was carried out.
Viral RNA extraction and viral RNA screening
All sand fly samples were ground in 200 μl of 1× PBS and then centrifuged at 11,000×g for 5 min. Subsequently, 100 μl of supernatant with 300 μl of minimum essential medium (MEM) was used for virus culture. The pellet and some of the supernatant with lysis buffer were processed for viral RNA extraction using the Invisorb Spin Virus RNA Mini Kit (STRATEC Molecular GmbH, Germany) following the manufacturer’s instructions. The extracted RNA samples were immediately used for detection of virus families, and the remaining samples were stored at −80 °C. The RNA samples were amplified for the detection of the Orbivirus , Phlebovirus , and Flavivirus  genera and family Rhabdoviridae . All the methods for viral RNA detection were briefly described in Phumee et al. . PCR amplicon sizes were determined by 1.5% ultrapure low-melting-point (LMP) agarose (Gibco, USA) gels, and gels were stained with ethidium bromide and visualized with Quantity One quantification analysis version 4.5.2 software (Gel Doc EQ System; Bio-Rad, USA). The estimated sizes for the Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae were 188, 244, 270, and 460 base pairs (bp), respectively. A synthesized plasmid was used as a positive control to prevent sample contamination.
One hundred microliters of each virus-positive suspension was inoculated onto confluent monolayers of Aedes albopictus C6/36 (ATCC CRL-1660) or rhesus monkey kidney epithelial (LLC-MK2) (ATCC CCL-7) cells for 1 h in 24-well plates grown in MEM (Gibco, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco, USA). The cultures were incubated at 28 °C (C6/36) and 37 °C (LLC-MK2) in 5% CO2. Cytopathic effects (CPEs) were monitored and observed daily for the following 6–10 days. The positive isolates were continually propagated in C6/36 cells and LLC-MK2 cells grown in MEM supplemented with 10% heat-inactivated FBS (Gibco, USA) at 28 °C (C6/36) and 37 °C (LLC-MK2) in a 5% CO2 environment for additional passages. The culture supernatants were harvested for identification and sequencing. Uninfected C6/36 or LLC-MK2 cells were used as negative controls.
Cloning and sequencing
PCR products were either directly sequenced or cloned into the pGEM-T Easy Vector (Promega, USA), transformed into DH5α competent cells, and screened by a blue–white colony selection system. The suspected positive colonies were cultured and used for further plasmid DNA extraction using the Invisorb Spin Plasmid Mini kit (STRATEC Molecular GmbH, Germany) following the manufacturer’s instructions. Purified plasmids were sent for sequencing by 1st BASE DNA sequencing services (1st BASE Laboratories, Malaysia) using universal forward T7 primers. Several independent PCR clones (3–5 clones) were analyzed to produce a consensus nucleotide sequence for each virus. Because of genetic variation within the degenerate primer sequences, they were excluded from the analysis. Nucleotide sequences were analyzed by comparing them with the reference viral sequence in the GenBank database using the BLASTn tool in the BLAST program. This tool searches nucleotide databases using percentage nucleotide queries and identity.
The nucleotide sequences were aligned using the BioEdit version 7.2.5 sequence alignment editor  and MAFFT (multiple alignment using fast Fourier transform) version 7 software (https://mafft.cbrc.jp/alignment/software/) . Phylogenetic trees of viruses were generated using both maximum likelihood (ML) and Bayesian approaches with IQ-TREE on the IQ-TREE web server (http://iqtree.cibiv.univie.ac.at/) with 1000 ultrafast bootstrap replicates. The best-fit model of substitution was identified using the auto function on the IQ-TREE web server . Finally, the phylogenetic tree was viewed and edited with FigTree version 1.4.4 (http://tree.bio.ed.ac.uk/software/figtree/).
This study is a descriptive analysis. The sample size was calculated following a simple formula  by estimated prevalence as a percentage from a pilot study and a previously published paper from our team , with 95% confidence. The data from the sand fly survey were calculated as the total number of sand fly species divided by the total population of the total number of all sand fly samples. The percentage of virus-sand fly infection was also calculated for the virus-positive sand flies, which was calculated by dividing the total number of positive samples of sand flies by the total number of all sand fly samples. All statistical analyses were performed using Microsoft Excel 2019 (Microsoft Corporation, USA).
A total of 331 specimens were analyzed. The samples included 162 (48.9%) sand flies collected in Trang and 169 (51.1%) sand flies collected in Songkhla. Phlebotomus stantoni and Se. indica were the most abundant sand fly species found in Trang and Songkhla provinces, respectively. Idiophlebotomus spp., Ph. mascomai, and Ph. papatasi were found only in Trang province. All samples were screened for viruses: Orbivirus, Phlebovirus, and Flavivirus genera and family Rhabdoviridae. The Orbivirus, Phlebovirus, and Flavivirus genera RT-PCRs were negative in all sand fly samples (Table 1). Interestingly, family Rhabdoviridae RNA was detected in three (0.9%) Ph. papatasi samples. Viral isolation was not successful in cell culture inoculation of the positive samples. No cytopathic effect (CPE) was observed in three positive sand fly samples, and culture supernatants remained negative for the family Rhabdoviridae. Rhabdoviridae family RNA was detected in three samples of Ph. papatasi. Sequencing of the L-region amplicon revealed a sequence length of approximately 450 bp. Sequences of all three samples were deposited in GenBank (accession nos. OK205076–OK205078). We observed that these three sequences had similarity of 74.16% compared to Bactrocera dorsalis sigmavirus strain BDSV.abc5 (accession no. MN745080), which has been reported as a new sigmavirus in the family Rhabdoviridae from fruit flies. The intramural diversity of three sequences and other sigmaviruses showed 19.2–41.5% nucleotide similarity. The ML analysis using the sequences from the three positives of family Rhabdoviridae in this study revealed a separate clustering from B. dorsalis sigmavirus strain BDSV.abc5 (accession no. MN745080), B. tryoni rhabdovirus 1 strain d (accession no. MW208811), rhabdoviruses or dimarhabdoviruses supergroup; vesiculovirus, Le Dantec and Kern Canyon group, Almpiwar group, sigmavirus, Hart Park group, and ephemerovirus (Fig. 1). Additionally, we conducted Bayesian analysis to investigate the correlation between the viruses and their host evolution by combining phylogenetic data with their host information. The results revealed that these three new sequences of sigmaviruses in Ph. papatasi from Thailand were categorized in a well-supported monophyletic group that was distinct from other dimarhabdoviruses (Fig. 2).
The study of pathogens in arthropods is an important part of surveillance programs. It can provide data about circulating pathogens and their vectors, assess potential public health threats, predict probable disease emergence in susceptible humans or animals, and establish optimal diagnostics and interventions to prevent disease transmission [30, 31]. In the present study, we detected family Rhabdoviridae RNA in three (0.9%) samples of Ph. papatasi sand flies from Trang Province. The Ph. papatasi was not found in Songkhla Province in this study. However, the results obtained did not reveal the circulation of the Orbivirus, Phlebovirus, or Flavivirus genera in field-caught sand flies. To the best of our knowledge, this is the first study undertaken to identify a new cluster of rhabdoviruses from Ph. papatasi in Thailand that was genetically closely related to B. dorsalis sigmavirus strain BDSV.abc5. Rhabdoviruses, a family of negative-sense RNA viruses (Mononegavirales: Rhabdoviridae), are a large and ecologically diverse group of viruses . For the ML phylogenetic tree, the sequences from this study showed separate clustering, exhibiting a close relationship to B. dorsalis sigmavirus strain BDSV.abc5 of the oriental fruit fly (accession no. MN745080)  and B. tryoni rhabdovirus 1 strain d of a tephritid fruit fly (accession no. MW208811) . The viruses have been reported as new sigmaviruses in the family Rhabdoviridae [33, 34]. Sigmavirus genera infects a variety of fruit flies, including Drosophila melanogaster, D. immigrans, and D. ananassae . It was found that flies infected with the sigmavirus of D. melanogaster (DMelSV) became paralyzed and died upon exposure to CO2, whereas uninfected flies recovered after exposure to CO2 . In addition, rhabdoviruses are involved in paralysis by binding to acetylcholine receptors on the nervous tissue of flies . Several reports suggested that rhabdovirus injection in aphids , Culex mosquitoes , and Drosophila  reduced their lifespan after CO2 exposure. From the aforementioned discussion, it is postulated that rhabdoviruses may exist in common groups of insects and may serve as common pathogens in those insect populations. Our findings demonstrated the first use of molecular techniques to detect the family Rhabdoviridae in sand flies from southern Thailand. The results revealed the presence of a new cluster of rhabdoviruses, which was closely related to the new sigmavirus in Ph. papatasi. The phylogeny of these sequences was reconstructed using Bayesian analysis with amino acid sequences and models of protein evolution. Our phylogeny showed that new sigmaviruses in Ph. papatasi are in the same lineage of dimarhabdoviruses, which suggested that infecting arthropod viruses emerged from this clade and might have undergone host switching during its evolution rather than cospeciation with them. Previous studies have revealed that for a pathogen to infect a new host species, it must adapt and adjust to the new environment in which it settles [40,41,42]. Additionally, the phylogenetic distance and root sharing between hosts also elucidate the diversity in virulence after cross-species transmission. Virulence increases when viruses jump between more distantly related hosts [43, 44]. Here, we report for the first time a new cluster of rhabdoviruses closely related to the new sigmavirus in sand flies from Thailand. Zhang et al.  implied that there was a strong association between sigmaviruses and their hosts. Therefore, the discovery of this virus may enrich our understanding of the diversity of RNA viruses in sand flies and provide information about virus association with their host and their adaptation in insects. This new sigmavirus may be a general rhabdovirus that is unlikely to cause high mortality. Unfortunately, the isolation of the virus in this study in Ae. albopictus C6/36 (ATCC CRL-1660) and rhesus monkey kidney epithelial (LLC-MK2) (ATCC CCL-7) cells was not successful, and the RT-PCR performed with the infected cells was negative. This likely resulted from a combination of factors associated with the C6/36 and LLC-MK2. Indeed, C6/36 cells have been used extensively for the isolation of rhabdoviruses [45, 46], but they may not be permissive to all insect viruses. In this regard, a non-specific cell line should be used instead. This may indicate that the culture of C6/36 or LLC-MK2 cells, although convenient, is be ideal. Few studies have shown different culture results from different cell types. A previous report in Italy showed the successful isolation of a novel rhabdovirus from an insectivorous bat (Pipistrellus kuhlii) in Vero cells . Another study showed that infecting LL-5 cells (sand fly cell line derived from sand fly Lutzomyia longipalpis) with vesicular stomatitis virus (VSV) from a sand fly could establish productive, long-lasting persistent infections. However, sustained persistence was difficult in C6/36 cells in the study [48,49,50]. Extensive studies aimed at identifying insect rhabdovirus vectors and their distribution in nature could help us understand the natural evolution of this virus; moreover, extensive surveys to demonstrate its presence in its natural vectors or host may have benefit in terms of demonstrating early signals of its circulation before emergence of diseases from such viruses and sand flies. Furthermore, the technical limitations of the present study based on conventional RT–PCR for viral screening, high-throughput experimental approaches, or metagenomic experimental design combined with the use of next-generation sequencing methods should be addressed in future studies.
This study reported a new cluster of rhabdoviruses closely related to the new sigmavirus in sand flies from southern Thailand. Additionally, the results also provide information on virus association with insect host adaptation. Although rhabdoviruses are already considered common pathogens in terms of insect-specific viruses, extensive surveys and comprehensive studies of new rhabdoviruses might be able to clarify and demonstrate the wide diversity of their host taxa as well as epidemiological data of this virus in sand flies. Moreover, further studies will be able to provide a better understanding of the relationships between rhabdoviruses and sand flies in Thailand.
Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Polymerase chain reaction
Reverse transcriptase PCR
Maroli M, Feliciangeli MD, Bichaud L, Charrel RN, Gradoni L. Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern. Med Vet Entomol. 2013;27:123–47.
Bates P, Depaquit J, Galati E, Kamhawi S, Maroli M, McDowell M, et al. Recent advances in phlebotomine sand fly research related to leishmaniasis control. Parasit Vectors. 2015;8:131.
Elaagip A, Ahmed A, Wilson MD, Boakye DA, Abdel Hamid MM. Studies of host preferences of wild-caught Phlebotomus orientalis and Ph. papatasi vectors of leishmaniasis in Sudan. PLoS ONE. 2020;15:e0236253.
Phumee A, Tawatsin A, Thavara U, Pengsakul T, Thammapalo S, Depaquit J, et al. Detection of an unknown Trypanosoma DNA in a Phlebotomus stantoni (Diptera: Psychodidae) collected from southern Thailand and records of new sand Flies with reinstatement of Sergentomyia hivernus Raynal & Gaschen, 1935 (Diptera: Psychodidae). J Med Entomol. 2017;54:429–34.
Srisuton P, Phumee A, Sunantaraporn S, Boonserm R, Sor-Suwan S, Brownell N, et al. Detection of Leishmania and Trypanosoma DNA in field-caught sand flies from endemic and non-endemic areas of leishmaniasis in southern Thailand. Insects. 2019;10:238.
Dionisio D, Esperti F, Vivarelli A, Valassina M. Epidemiological, clinical and laboratory aspects of sandfly fever. Curr Opin Infect Dis. 2003;16:383–8.
Tavana AM. Sandfly fever in the world. Ann Trop Med Public Health. 2015;8:83–7.
Tesh RB. The genus Phlebovirus and its vectors. Annu Rev Entomol. 1988;33:169–81.
Marklewitz M, Tchouassi DP, Hieke C, Heyde V, Torto B, Sang R, et al. Insights into the evolutionary origin of Mediterranean sandfly fever viruses. mSphere. 2020;5:e00598-e620.
Bhatt PN, Rodrigues FM. Chandipura: a new arbovirus isolated in India from patients with febrile illness 1967. Indian J Med Res. 2012;136:13–890.
Silva SP, Dilcher M, Weber F, Hufert FT, Weidmann M, Cardoso JF, et al. Genetic and biological characterization of selected Changuinola viruses (Reoviridae, Orbivirus) from Brazil. J Gen Virol. 2014;95:2251–9.
Moureau G, Ninove L, Izri A, Cook S, De Lamballerie X, Charrel RN. Flavivirus RNA in phlebotomine sandflies. Vector Borne Zoonotic Dis. 2010;10:195–7.
Fu ZF. Genetic comparison of the rhabdoviruses from animals and plants. Curr Top Microbiol Immunol. 2005;292:1–24.
Afonso CL, Amarasinghe GK, Bányai K, Bào Y, Basler CF, Bavari S, et al. Taxonomy of the order Mononegavirales: update 2016. Arch Virol. 2016;161:2351–60.
Adams MJ, Lefkowitz EJ, King AM, Harrach B, Harrison RL, Knowles NJ, et al. Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2016). Arch Virol. 2016;161:2921–49.
Longdon B, Obbard DJ, Jiggins FM. Sigma viruses from three species of Drosophila form a major new clade in the rhabdovirus phylogeny. Proc Biol Sci. 2010;277:35–44.
Rocklöv J, Dubrow R. Climate change: an enduring challenge for vector-borne disease prevention and control. Nat Immunol. 2020;21:479–83.
Phumee A, Wacharapluesadee S, Petcharat S, Tawatsin A, Thavara U, Siriyasatien P. Detection of Changuinola virus (Reoviridae: Orbivirus) in field-caught sand flies in southern Thailand. Trans R Soc Trop Med Hyg. 2021;115:1039-44.
Lewis DJ. The phlebotomine sand flies (Diptera: Psychodidae) of the oriental region. Bull Br MUS Nat Hist. 1978;37:217–343.
Depaquit J, Vongphayloth K, Siriyasatien P, Polseela R, Phumee A, Loyer M, et al. On the true identity of Sergentomyia gemmea and description of a closely related species: Se. raynali n. sp. Med Vet Entomol. 2019;33:521–9.
Esseghir S, Ready PD, Killick-Kendrick R, Ben-Ismail R. Mitochondrial haplotypes and phylogeography of Phlebotomus vectors of Leishmania major. Insect Mol Biol. 1997;6:211–25.
Palacios G, Cowled C, Bussetti AV, Savji N, Weir R, Wick I, et al. Rapid molecular strategy for orbivirus detection and characterization. J Clin Microbiol. 2011;49:2314–7.
Sánchez-Seco MP, Echevarría JM, Hernández L, Estévez D, Navarro-Marí JM, Tenorio A. Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers. J Med Virol. 2003;71:140–9.
Moureau G, Temmam S, Gonzalez JP, Charrel RN, Grard G, de Lamballerie X. A real-time RT-PCR method for the universal detection and identification of flaviviruses. Vector Borne Zoonotic Dis. 2007;7:467–77.
Bourhy H, Cowley JA, Larrous F, Holmes EC, Walker PJ. Phylogenetic relationships among rhabdoviruses inferred using the L polymerase gene. J Gen Virol. 2005;86:2849–58.
Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucl Acids Symp Ser. 1999;41:95–8.
Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol. 2013;30:772–80.
Trifinopoulos J, Nguyen LT, von Haeseler A, Minh BQ. W-IQ-TREE: a fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Res. 2016;44:W232–5.
Daniel WW. Biostatistics: a foundation for analysis in the health sciences. 7th ed. New York: Wiley; 1999.
Medlock JM, Hansford KM, Van Bortel W, Zeller H, Alten B. A summary of the evidence for the change in European distribution of phlebotomine sand flies (Diptera: Psychodidae) of public health importance. J Vector Ecol. 2014;39:72–7.
Liang G, Gao X, Gould EA. Factors responsible for the emergence of arboviruses; strategies, challenges and limitations for their control. Emerg Microbes Infect. 2015;4:e18.
Dietzgen RG, Kondo H, Goodin MM, Kurath G, Vasilakis N. The family Rhabdoviridae: mono- and bipartite negative-sense RNA viruses with diverse genome organization and common evolutionary origins. Virus Res. 2017;227:158–70.
Zhang W, Gu Q, Niu J, Wang JJ. The RNA virome and its dynamics in an invasive fruit fly, Bactrocera dorsalis, imply interactions between host and viruses. Microb Ecol. 2020;80:423–34.
Sharpe SR, Morrow JL, Brettell LE, Shearman DC, Gilchrist S, Cook JM, Riegler M. Tephritid fruit flies have a large diversity of co-occurring RNA viruses. J Invertebr Pathol. 2021. https://doi.org/10.1016/j.jip.2021.107569.
Webster CL, Longdon B, Lewis SH, Obbard DJ. Twenty-five new viruses associated with the Drosophilidae (Diptera). Evol Bioinform Online. 2016;12:13–25.
Longdon B, Wilfert L, Obbard DJ, Jiggins FM. Rhabdoviruses in two species of Drosophila: vertical transmission and a recent sweep. Genetics. 2011;188:141–50.
Brun G, Plus N. The viruses of Drosophila. In: Ashburner M, Wright TRF, editors. The genetics and biology of Drosophila. New York: Academic Press; 1980. p. 625–702.
Sylvester E, Richardson J. Aphid-borne rhabdoviruses-relationship with their aphid vectors. In: Harris KF, editor. Advances disease vector research. New York: Springer; 1992. p. 313–41.
Shroyer DA, Rosen L. Extrachromosomal inheritance of carbon dioxide sensitivity in the mosquito Culex quinquefasciatus. Genetics. 1983;104:649–59.
Davies TJ, Pedersen AB. Phylogeny and geography predict pathogen community similarity in wild primates and humans. Proc Biol Sci. 2008;275:1695–701.
Parrish CR, Holmes EC, Morens DM, Park EC, Burke DS, Calisher CH, et al. Cross-species virus transmission and the emergence of new epidemic diseases. Microbiol Mol Biol Rev. 2008;72:457–70.
Woolhouse ME, Haydon DT, Antia R. Emerging pathogens: the epidemiology and evolution of species jumps. Trends Ecol Evol. 2005;20:238–44.
Guth S, Visher E, Boots M, Brook CE. Host phylogenetic distance drives trends in virus virulence and transmissibility across the animal-human interface. Philos Trans R Soc Lond B Biol Sci. 2019;374:20190296.
Mollentze N, Streicker DG, Murcia PR, Hampson K, Biek R. Virulence mismatches in index hosts shape the outcomes of cross-species transmission. Proc Natl Acad Sci USA. 2020;117:28859–66.
Vazeille MC, Rosen L. Preparation of high-titred antisera for sigma virus strains from Hawaii and France and their use in neutralization tests in vitro and in vivo. Ann Inst Pasteur Virol. 1988;139:175–83.
Vazeille MC, Rosen L. Cultivation in mosquito cells of sigma virus, the aetiologic agent of hereditary CO2 sensitivity in Drosophila melanogaster. Ann Inst Pasteur Virol. 1987;138:451–60.
Lelli D, Prosperi A, Moreno A, Chiapponi C, Gibellini AM, De Benedictis P, et al. Isolation of a novel Rhabdovirus from an insectivorous bat (Pipistrellus kuhlii) in Italy. Virol J. 2018;15:37.
Tesh RB, Modi GB. Development of a continuous cell line from the sand fly Lutzomyia longipalpis (Diptera: Psychodidae), and its susceptibility to infection with arboviruses. J Med Entomol. 1983;20:199–202.
Llewellyn ZN, Salman MD, Pauszek S, Rodriguez LL. Growth and molecular evolution of vesicular stomatitis serotype New Jersey in cells derived from its natural insect-host: evidence for natural adaptation. Virus Res. 2002;89:65–73.
Zárate S, Novella IS. Vesicular stomatitis virus evolution during alternation between persistent infection in insect cells and acute infection in mammalian cells is dominated by the persistence phase. J Virol. 2004;78:12236–42.
We would like to thank the new strategic research project (P2P), Walailak University, Thailand (Grant No. CGS-P2P-2564-012), the Research Institute for Health Sciences, Walailak University (Grant No. WU-IRG-64-013), the Program Management Unit for Human Resources and Institutional Development, Research and Innovation-CU (Grant No. B16F630071), Thailand Science Research Innovation (TSRI)-CU (Grant No. FRB640001), and NIH/NIAID/CREID/07-049-7012-52338 for support.
This study was supported by the new strategic research project (P2P), Walailak University, Thailand (Grant No. CGS-P2P-2564-012), the Research Institute for Health Sciences, Walailak University (Grant No. WU-IRG-64-013), the Program Management Unit for Human Resources and Institutional Development, Research and Innovation-CU (Grant No. B16F630071), Thailand Science Research Innovation (TSRI)-CU (Grant No. FRB640001), and NIH/NIAID/CREID/07-049-7012-52338.
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The study was approved by the Animal Ethical Committee of Walailak University (Clearance No. WU-AICUC-64-005).
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Phumee, A., Wacharapluesadee, S., Petcharat, S. et al. A new cluster of rhabdovirus detected in field-caught sand flies (Diptera: Psychodidae: Phlebotominae) collected from southern Thailand. Parasites Vectors 14, 569 (2021). https://doi.org/10.1186/s13071-021-05047-z
- Molecular survey
- Sand fly