Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
PLoS Negl Trop Dis
2021 Dec 20;1512:e0009942. doi: 10.1371/journal.pntd.0009942.
Show Gene links
Show Anatomy links
Molecular phylogeny of heritable symbionts and microbiota diversity analysis in phlebotominae sand flies and Culex nigripalpus from Colombia.
Vivero-Gomez RJ
,
Castañeda-Monsalve VA
,
Atencia MC
,
Hoyos-Lopez R
,
Hurst GD
,
Cadavid-Restrepo G
,
Moreno-Herrera CX
.
???displayArticle.abstract???
BACKGROUND: Secondary symbionts of insects include a range of bacteria and fungi that perform various functional roles on their hosts, such as fitness, tolerance to heat stress, susceptibility to insecticides and effects on reproduction. These endosymbionts could have the potential to shape microbial communites and high potential to develop strategies for mosquito-borne disease control.
METHODOLOGY/PRINCIPAL FINDINGS: The relative frequency and molecular phylogeny of Wolbachia, Microsporidia and Cardinium were determined of phlebotomine sand flies and mosquitoes in two regions from Colombia. Illumina Miseq using the 16S rRNA gene as a biomarker was conducted to examine the microbiota. Different percentages of natural infection by Wolbachia, Cardinium, and Microsporidia in phlebotomines and mosquitoes were detected. Phylogenetic analysis of Wolbachia shows putative new strains of Lutzomyia gomezi (wLgom), Brumptomyia hamata (wBrham), and a putative new group associated with Culex nigripalpus (Cnig) from the Andean region, located in Supergroup A and Supergroup B, respectively. The sequences of Microsporidia were obtained of Pi. pia and Cx. nigripalpus, which are located on phylogeny in the IV clade (terrestrial origin). The Cardinium of Tr. triramula and Ps. shannoni were located in group C next to Culicoides sequences while Cardinium of Mi. cayennensis formed two putative new subgroups of Cardinium in group A. In total were obtained 550 bacterial amplicon sequence variants (ASVs) and 189 taxa to the genus level. The microbiota profiles of Sand flies and mosquitoes showed mainly at the phylum level to Proteobacteria (67.6%), Firmicutes (17.9%) and Actinobacteria (7.4%). High percentages of relative abundance for Wolbachia (30%-83%) in Lu. gomezi, Ev. dubitans, Mi. micropyga, Br. hamata, and Cx. nigripalpus were found. ASVs assigned as Microsporidia were found in greater abundance in Pi. pia (23%) and Cx. nigripalpus (11%). An important finding is the detection of Rickettsia in Pi. pia (58,8%) and Bartonella sp. in Cx. nigripalpus.
CONCLUSIONS/SIGNIFICANCE: We found that Wolbachia infection significantly decreased the alpha diversity and negatively impacts the number of taxa on sand flies and Culex nigripalpus. The Principal Coordinate Analysis (PCoA) is consistent, which showed statistically significant differences (PERMANOVA, F = 2.4744; R2 = 0.18363; p-value = 0.007) between the microbiota of sand flies and mosquitoes depending on its origin, host and possibly for the abundance of some endosymbionts (Wolbachia, Rickettsia).
Fig 1. Geographic locations of sampling sites and proportion by department of insect vectors (Phlebotomine sand flies and mosquitoes) infected with Wolbachia, Cardinium and Microsporidia in Colombia.Link to the base layer of the map used: https://www.colombiaenmapas.gov.co/?e=-74.7117818345732,0.23307652371168444,-62.73668417832639,8.735414279337139,4686&b=dark-gray&u=0&t=2901&servicio=610.
Fig 2. Phylogenetic relationships of Wolbachia strains using Bayesian analysis and inferred with wsp sequences.The Wolbachia strains are identified by host species from which they were isolated (sand fllies symbol of green color; Culex symbol of orange color). Numbers in nodes represent Bayesian posterior probabilities. Reconstruction performed with MrBayes (version 3.0). The wMel positive control is indicated in red color. As out-group Wolbachia of Bemicia tabaci.
Fig 3. Bayesian inference and phylogenetic relationships of Microsporidian strains using sequences of the small subunit ribosomal DNA.The Microsporidian strains are identified by host species from which they were isolated (sand fly symbol of green color; Culex nigripalpus of orange color). Numbers in nodes represent Bayesian posterior probabilities. Reconstruction performed with MrBayes (version 3.0). Clades I–V are indicated. Two fungi were used as outgroups in the analysis (Conidiobolus coronatos and Basidiobolus ranarun). Taxa without GenBank accession numbers are original for this dataset.
Fig 4. Phylogenetic relationships of Cardinium strains using Bayesian analysis and based on 16S rDNA sequences.The Cardinium strains are identified by host species from which they were isolated (sand flies symbol of green color). Numbers in nodes represent Bayesian posterior probabilities. The reconstruction was performed using MrBayes (version 3.0). As an outgroup was used Candidatus Amoebophilus asiaticus. Taxa without GenBank accession numbers are original for this data set.
Fig 5. Neighbor-joining analysis of mitochondrial COI sequences among sand flies species (symbol of green color) and mosquitoes of genus Culex (symbol of orange color) from Colombia, based on the model of Kimura’s two-parameter model (K2P).Bootstrap support was estimated with 1000 replicas, only bootstrap values >50% being shown above the branch. As an outgroup was used Tipula maxima.
Fig 6. Microbiota composition and diversity of wild specimens of sand flies and Culex nigripalpus found with endosymbiont in several regions or departments from Colombia.A) Relative abundance of ASVs that were called to the taxonomic rank of genus. The ASVs with abundances above 1% of the rarefied count matrix were selected. See Table 4 for the nomenclature of samples in detail. NA: ASVs without taxonomic assignment.
Fig 7. Alpha diversity (intra-diversity) of bacterial composition using amplicon sequence variants (ASVs) in sand flies and Cx. nigripalpus from Colombia by ecoregion and presence of endosymbiont.a) Species richness was assessed by Chao1 (estimates diversity from abundance data), b) Observed OTU (sum of unique OTUs in each sample), c) Shannon-Wiener (sum of species proportion logarithms) and d) Simpson indexes. e) Alpha diversity with all indexes but associated with the presence of endosymbionts. Higher index values indicate richer and more diverse bacterial composition.
Fig 8. β-diversity analysis (between groups) of microbial communities associated with the groups of samples established of sand flies and Culex nigripalpus found with endosymbiont in several ecoregions from Colombia.a) Hierarchical clustering analysis of ASVs at the genus level. b) Principal Coordinate Analysis (PCoA) with ASVs at the genus level (Weighted Unifrac Distance, PERMANOVA, F = 2.4744; R2 = 0.18363; p-value = 0.007). c) Beta-diversity visualized using Non-metric Multidimensional Scaling (NMDS). The plot was obtained with Bray-Curtis Dissimilarity distances (PERMANOVA, F = 1.1429; R2 = 0.094118; p-value = 0.324; [NMDS] Stress = 0.11348).
Fig 9. Heatmap based on the microbiota composition at the genus level illustrating common and endemic bacterial community abundances according to insects, the presence of endosymbionts and categorization according to its importance as a vector.Hierarchical Ward’s linkage clustering based on Pearson’s correlation coefficient of the microbial taxa abundance. Red and blue colors represent positive and negative correlations, respectively. The color scale represents the scaled abundance of each variable, denoted as Z-score, red indicates high abundance, and blue indicating low abundance.
Fig 10. Linear discriminant analysis effect size (LEfSe) of gut microbiota in sand flies and Culex nigripalpus de Colombia.
Fig 11. The random forest analysis shows positive interactions and significant features among some taxa in the core microbiota in sand flies and Culex nigripalpus de Colombia.The top 25 of ASVs most abundant are considered. The y axis, from top to bottom, displays the taxa ranked by their importance or association with other ASV and ecoregion.
Abdul Rahman,
A molecular survey of Australian and North American termite genera indicates that vertical inheritance is the primary force shaping termite gut microbiomes.
2015, Pubmed
Abdul Rahman,
A molecular survey of Australian and North American termite genera indicates that vertical inheritance is the primary force shaping termite gut microbiomes.
2015,
Pubmed
Alexander,
A review of bartonellosis in Ecuador and Colombia.
1995,
Pubmed
Altamiranda-Saavedra,
Downscaling incidence risk mapping for a Colombian malaria endemic region.
2018,
Pubmed
Altamiranda-Saavedra,
Effect of El Niño Southern Oscillation cycle on the potential distribution of cutaneous leishmaniasis vector species in Colombia.
2020,
Pubmed
Anbutsu,
Spiroplasma as a model insect endosymbiont.
2011,
Pubmed
Andreadis,
Microsporidian parasites of mosquitoes.
2007,
Pubmed
Araújo-Coutinho,
Seasonality and prevalence rates of microsporidia in Simulium pertinax (Diptera: Simuliidae) larvae in the region of Serra dos Orgãos, Rio de Janeiro, Brasil.
2004,
Pubmed
Azpurua,
Lutzomyia sand fly diversity and rates of infection by Wolbachia and an exotic Leishmania species on Barro Colorado Island, Panama.
2010,
Pubmed
Baldini,
First report of natural Wolbachia infection in the malaria mosquito Anopheles arabiensis in Tanzania.
2018,
Pubmed
Barua,
First report of Rickettsia felis in mosquitoes, USA.
2020,
Pubmed
Bascuñán,
Factors shaping the gut bacterial community assembly in two main Colombian malaria vectors.
2018,
Pubmed
Bolaños,
Arthropod-Spiroplasma relationship in the genomic era.
2015,
Pubmed
Braig,
Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis.
1998,
Pubmed
Brown,
Comparative Genomics of Wolbachia-Cardinium Dual Endosymbiosis in a Plant-Parasitic Nematode.
2018,
Pubmed
Callahan,
DADA2: High-resolution sample inference from Illumina amplicon data.
2016,
Pubmed
Caporaso,
Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms.
2012,
Pubmed
Carvalho,
Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae).
2014,
Pubmed
Charlesworth,
Wolbachia, Cardinium and climate: an analysis of global data.
2019,
Pubmed
Chepkemoi,
Identification of Spiroplasmainsolitum symbionts in Anopheles gambiae.
2017,
Pubmed
Chong,
Using MicrobiomeAnalyst for comprehensive statistical, functional, and meta-analysis of microbiome data.
2020,
Pubmed
Contreras Gutiérrez,
DNA barcoding for the identification of sand fly species (Diptera, Psychodidae, Phlebotominae) in Colombia.
2014,
Pubmed
Dale,
Molecular interactions between bacterial symbionts and their hosts.
2006,
Pubmed
Day,
Factors That Influence the Transmission of West Nile Virus in Florida.
2015,
Pubmed
Dobson,
Wolbachia infections are distributed throughout insect somatic and germ line tissues.
1999,
Pubmed
Doremus,
Exposure to opposing temperature extremes causes comparable effects on Cardinium density but contrasting effects on Cardinium-induced cytoplasmic incompatibility.
2019,
Pubmed
Duguma,
Microbiota variations in Culex nigripalpus disease vector mosquito of West Nile virus and Saint Louis Encephalitis from different geographic origins.
2019,
Pubmed
Duron,
The diversity of reproductive parasites among arthropods: Wolbachia do not walk alone.
2008,
Pubmed
Fang,
Competitive ability and fitness differences between two introduced populations of the invasive whitefly Bemisia tabaci Q in China.
2014,
Pubmed
Folmer,
DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates.
1994,
Pubmed
,
Echinobase
Fraga,
Phylogeny of Leishmania species based on the heat-shock protein 70 gene.
2010,
Pubmed
Fredensborg,
Parasites modulate the gut-microbiome in insects: A proof-of-concept study.
2020,
Pubmed
Gasparich,
The genus Spiroplasma and its non-helical descendants: phylogenetic classification, correlation with phenotype and roots of the Mycoplasma mycoides clade.
2004,
Pubmed
Glaser,
The native Wolbachia endosymbionts of Drosophila melanogaster and Culex quinquefasciatus increase host resistance to West Nile virus infection.
2010,
Pubmed
Gotoh,
Wolbachia-induced cytoplasmic incompatibility in Japanese populations of Tetranychus urticae (Acari: Tetranychidae).
2007,
Pubmed
Gubry-Rangin,
Coupling of diversification and pH adaptation during the evolution of terrestrial Thaumarchaeota.
2015,
Pubmed
Guerrero,
Symbiogenesis: the holobiont as a unit of evolution.
2013,
Pubmed
Hanahan,
Studies on transformation of Escherichia coli with plasmids.
1983,
Pubmed
Harris,
Population dynamics and rapid spread of Cardinium, a bacterial endosymbiont causing cytoplasmic incompatibility in Encarsia pergandiella (Hymenoptera: Aphelinidae).
2010,
Pubmed
Hebert,
Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species.
2003,
Pubmed
Herren,
A microsporidian impairs Plasmodium falciparum transmission in Anopheles arabiensis mosquitoes.
2020,
Pubmed
Himler,
Rapid spread of a bacterial symbiont in an invasive whitefly is driven by fitness benefits and female bias.
2011,
Pubmed
Hoyos-López,
Molecular detection of flaviviruses and alphaviruses in mosquitoes (Diptera: Culicidae) from coastal ecosystems in the Colombian Caribbean.
2016,
Pubmed
Jaenike,
Adaptation via symbiosis: recent spread of a Drosophila defensive symbiont.
2010,
Pubmed
Jeffries,
Establishment of a method for Lutzomyia longipalpis sand fly embryo microinjection: The first step towards potential novel control strategies for leishmaniasis.
2018,
Pubmed
Jeyaprakash,
Long PCR improves Wolbachia DNA amplification: wsp sequences found in 76% of sixty-three arthropod species.
2000,
Pubmed
Karatepe,
Investigation of Wolbachia spp. and Spiroplasma spp. in Phlebotomus species by molecular methods.
2018,
Pubmed
Karimian,
wsp-based analysis of Wolbachia strains associated with Phlebotomus papatasi and P. sergenti (Diptera: Psychodidae) main cutaneous leishmaniasis vectors, introduction of a new subgroup wSerg.
2018,
Pubmed
Keeling,
Microsporidia: biology and evolution of highly reduced intracellular parasites.
2002,
Pubmed
Konecka,
First Evidence of Intracellular Bacteria Cardinium in Thermophilic Mite Microzetorchestes emeryi (Acari: Oribatida): Molecular Screening of Bacterial Endosymbiont Species.
2019,
Pubmed
Kontsedalov,
The presence of Rickettsia is associated with increased susceptibility of Bemisia tabaci (Homoptera: Aleyrodidae) to insecticides.
2008,
Pubmed
Kumar,
MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.
2018,
Pubmed
Li,
Diversity of bacteriome associated with Phlebotomus chinensis (Diptera: Psychodidae) sand flies in two wild populations from China.
2016,
Pubmed
Libertucci,
The role of the microbiota in infectious diseases.
2019,
Pubmed
Librado,
DnaSP v5: a software for comprehensive analysis of DNA polymorphism data.
2009,
Pubmed
Louradour,
The midgut microbiota plays an essential role in sand fly vector competence for Leishmania major.
2017,
Pubmed
Lozano-Sardaneta,
First Report of Bartonella SP. In Sand Flies (Diptera: Psychodidae: Phlebotominae) From Southern Mexico.
2019,
Pubmed
Mancini,
Wolbachia strain wAu efficiently blocks arbovirus transmission in Aedes albopictus.
2020,
Pubmed
Marcon,
Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae).
2011,
Pubmed
Martin,
Infections with the microbe Cardinium in the Dolichopodidae and other Empidoidea.
2013,
Pubmed
Martin,
RDP4: Detection and analysis of recombination patterns in virus genomes.
2015,
Pubmed
Matos,
Vavraia lutzomyiae n. sp. (Phylum Microspora) infecting the sandfly Lutzomyia longipalpis (Psychodidae, Phlebotominae), a vector of human visceral leishmaniasis.
2006,
Pubmed
McMurdie,
phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data.
2013,
Pubmed
Mee,
Detection of Low-Level Cardinium and Wolbachia Infections in Culicoides.
2015,
Pubmed
Merçot,
Variability within the Seychelles cytoplasmic incompatibility system in Drosophila simulans.
1995,
Pubmed
Moreira,
A Wolbachia symbiont in Aedes aegypti limits infection with dengue, Chikungunya, and Plasmodium.
2009,
Pubmed
Morningstar,
Diversity of Wolbachia pipientis strain wPip in a genetically admixtured, above-ground Culex pipiens (Diptera: Culicidae) population: association with form molestus ancestry and host selection patterns.
2012,
Pubmed
Morçiçek,
Evidence of natural Wolbachia infections and molecular identification of field populations of Culex pipiens complex (Diptera: Culicidae) mosquitoes in western Turkey.
2018,
Pubmed
Nakamura,
Prevalence of Cardinium bacteria in planthoppers and spider mites and taxonomic revision of "Candidatus Cardinium hertigii" based on detection of a new Cardinium group from biting midges.
2009,
Pubmed
Novakova,
Mosquito Microbiome Dynamics, a Background for Prevalence and Seasonality of West Nile Virus.
2017,
Pubmed
Ono,
Wolbachia infections of phlebotomine sand flies (Diptera: Psychodidae).
2001,
Pubmed
Papadopoulos,
Host Species Determines the Composition of the Prokaryotic Microbiota in Phlebotomus Sandflies.
2020,
Pubmed
Perlmutter,
Microorganisms in the reproductive tissues of arthropods.
2020,
Pubmed
Perrot-Minnot,
Single and double infections with Wolbachia in the parasitic wasp Nasonia vitripennis: effects on compatibility.
1996,
Pubmed
Pietri,
The rich somatic life of Wolbachia.
2016,
Pubmed
Quintero V,
Eco-epidemiological analysis of rickettsial seropositivity in rural areas of Colombia: A multilevel approach.
2017,
Pubmed
Raina,
Infection of Bacterial Endosymbionts in Insects: A Comparative Study of Two Techniques viz PCR and FISH for Detection and Localization of Symbionts in Whitefly, Bemisia tabaci.
2015,
Pubmed
Rath,
Uncovering the trimethylamine-producing bacteria of the human gut microbiota.
2017,
Pubmed
Reeves,
Pathogen screening and bionomics of Lutzomyia apache (Diptera: Psychodidae) in Wyoming, USA.
2008,
Pubmed
Ronquist,
MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space.
2012,
Pubmed
Rudolf,
Bartonella species in medically important mosquitoes, Central Europe.
2020,
Pubmed
Salgado-Almario,
Geographical distribution of Leishmania species in Colombia, 1985-2017.
2019,
Pubmed
Santos-Garcia,
The genome of Cardinium cBtQ1 provides insights into genome reduction, symbiont motility, and its settlement in Bemisia tabaci.
2014,
Pubmed
Shaw,
Wolbachia infections in natural Anopheles populations affect egg laying and negatively correlate with Plasmodium development.
2016,
Pubmed
Shokal,
Effects of co-occurring Wolbachia and Spiroplasma endosymbionts on the Drosophila immune response against insect pathogenic and non-pathogenic bacteria.
2016,
Pubmed
Sinkins,
Wolbachia superinfections and the expression of cytoplasmic incompatibility.
1995,
Pubmed
Socolovschi,
Rickettsia felis in Aedes albopictus mosquitoes, Libreville, Gabon.
2012,
Pubmed
Staley,
The bacterial species dilemma and the genomic-phylogenetic species concept.
2006,
Pubmed
Tolley,
Wolbachia Horizontal Transmission Events in Ants: What Do We Know and What Can We Learn?
2019,
Pubmed
Vivero,
Molecular detection and identification of Wolbachia in three species of the genus Lutzomyia on the Colombian Caribbean coast.
2017,
Pubmed
Vivero,
Wild specimens of sand fly phlebotomine Lutzomyia evansi, vector of leishmaniasis, show high abundance of Methylobacterium and natural carriage of Wolbachia and Cardinium types in the midgut microbiome.
2019,
Pubmed
Vossbrinck,
Verification of intermediate hosts in the life cycles of microsporidia by small subunit rDNA sequencing.
1998,
Pubmed
Vossbrinck,
Molecular phylogeny of the Microsporidia: ecological, ultrastructural and taxonomic considerations.
2005,
Pubmed
Vossbrinck,
Molecular phylogeny and evolution of mosquito parasitic Microsporidia (Microsporidia: Amblyosporidae).
2004,
Pubmed
Wang,
Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy.
2007,
Pubmed
Wasala,
Variable Abundance and Distribution of Wolbachia and Cardinium Endosymbionts in Plant-Parasitic Nematode Field Populations.
2019,
Pubmed
Weeks,
Incidence of a new sex-ratio-distorting endosymbiotic bacterium among arthropods.
2003,
Pubmed
Ye,
The Effect of Temperature on Wolbachia-Mediated Dengue Virus Blocking in Aedes aegypti.
2016,
Pubmed
Yen,
New hypothesis of the cause of cytoplasmic incompatibility in Culex pipiens L.
1971,
Pubmed
Zha,
Detection and characterization of Wolbachia infection in silkworm.
2014,
Pubmed
Zhang,
Rickettsia in mosquitoes, Yangzhou, China.
2016,
Pubmed
Zhou,
Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences.
1998,
Pubmed
da Silva Gonçalves,
Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum.
2019,
Pubmed
de Oliveira,
Broader prevalence of Wolbachia in insects including potential human disease vectors.
2015,
Pubmed
