The midgut microbiota plays an essential role in sand fly vector competence for<i>Leishmania major</i>

Louradour, Isabelle; Monteiro, Carolina Cunha; Inbar, Ehud; Ghosh, Kashinath; Merkhofer, Richard M.; Lawyer, Phillip G.; Paun, Andrea; Smelkinson, Margery; Secundino, Nágila Francinete Costa; Lewis, Michael D.; Erram, Dinesh; Zurek, Ludek; Sacks, David L.

doi:10.1111/cmi.12755

Cited by 72 publications

(64 citation statements)

References 38 publications

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“…Considerable variation was observed in the composition of the microbiota within each group (Fig. S5 and S6); however, this variability was also detected between different groups of wt flies, consistent with our previous findings (34). In order to compare the quantities of bacteria colonizing the guts of rel mutant and wt flies during Leishmania infection, we performed 16S quantitative PCR (qPCR) experiments on pooled guts of females infected with L. major.…”

Section: Resultssupporting

confidence: 87%

“…The gut microbiota is modified in rel mutant flies. Previous studies reported that the sand fly gut microbiota plays a crucial role in its vector competence for Leishmania (33,34). With this in mind, we compared the relative abundances of bacterial species between small groups of wt and rel B heterozygous or homozygous mutants in the absence of Leishmania infection.…”

Section: Resultsmentioning

confidence: 99%

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CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

Louradour

Ghosh

Inbar

et al. 2019

mBio

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Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report, we adapted CRISPR (clustered regularly interspaced palindromic repeat)/Cas9 (CRISPR-associated protein 9) technology to the Phlebotomus papatasi sand fly, a natural vector for Leishmania major, targeting the sand fly immune deficiency (IMD) pathway in order to decipher its contribution to vector competence. We established a protocol for transformation in P. papatasi and were able to generate transmissible null mutant alleles for Relish (Rel), the only transcription factor of the IMD pathway. While the maintenance of a homozygous mutant stock was severely compromised, we were able to establish in an early generation their greater susceptibility to infection with L. major. Flies carrying different heterozygous mutant alleles variably displayed a more permissive phenotype, presenting higher loads of parasites or greater numbers of infective-stage promastigotes. Together, our data show (i) the successful adaptation of the CRISPR/Cas9 technology to sand flies and (ii) the impact of the sand fly immune response on vector competence for Leishmania parasites. IMPORTANCE Sand flies are the natural vectors of Leishmania parasites. Studies of sand fly/Leishmania interactions have been limited by the lack of successful genomic manipulation of these insects. This paper shows the first example of successful targeted mutagenesis in sand flies via adaptation of the CRISPR/Cas9 editing technique. We generated transmissible null mutant alleles of relish, a gene known to be essential for the control of immune response in other insects. In addition to the expected higher level of susceptibility to bacteria, the mutant flies presented higher loads of parasites when infected with L. major, showing that the sand fly immune response impacts its vector competence for this pathogen.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

Louradour

Ghosh

Inbar

et al. 2019

mBio

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“…Phytocannabinoids from C. sativa exhibited antimicrobial activity against some bacteria and fungi (33). Since thriving gut microbiomes are crucial for the development of Leishmania infections in the sand fly gut (34,35), a microbicidal effect could harm Leishmania infections. Lastly, partially characterized compounds from C. sativa exhibited potent antileishmanial activity in vitro (36).…”

Section: Resultsmentioning

confidence: 99%

Plant-feeding phlebotomine sand flies, vectors of leishmaniasis, prefer Cannabis sativa

Abbasi

Queiroz

Kirstein

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Blood-sucking phlebotomine sand flies (Diptera: Psychodidae) transmit leishmaniasis as well as arboviral diseases and bartonellosis. Sand fly females become infected with Leishmania parasites and transmit them while imbibing vertebrates’ blood, required as a source of protein for maturation of eggs. In addition, both females and males consume plant-derived sugar meals as a source of energy. Plant meals may comprise sugary solutions such as nectar or honeydew (secreted by plant-sucking homopteran insects), as well as phloem sap that sand flies obtain by piercing leaves and stems with their needle-like mouthparts. Hence, the structure of plant communities can influence the distribution and epidemiology of leishmaniasis. We designed a next-generation sequencing (NGS)–based assay for determining the source of sand fly plant meals, based upon the chloroplast DNA gene ribulose bisphosphate carboxylase large chain (rbcL). Here, we report on the predilection of several sand fly species, vectors of leishmaniasis in different parts of the world, for feeding on Cannabis sativa. We infer this preference based on the substantial percentage of sand flies that had fed on C. sativa plants despite the apparent “absence” of these plants from most of the field sites. We discuss the conceivable implications of the affinity of sand flies for C. sativa on their vectorial capacity for Leishmania and the putative exploitation of their attraction to C. sativa for the control of sand fly-borne diseases.

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“…Furthermore, the gut bacteria of hematophagous vectors interact with parasites (like T. cruzi) occupying the same niche [2,23,24]. The microbiome can potentially impede parasite transmission through direct (competition for resources) and indirect (promoting immune response) interactions [25][26][27][28][29][30][31]. The most comprehensive background on hematophagous microbiomes has been derived from mosquitos, ticks, and tsetse flies (reviewed in [32]), whereas triatomine-bacteria associations remain neglected.…”

Section: Introductionmentioning

confidence: 99%

Ontogeny, species identity and environment dominate microbiome dynamics in wild populations of kissing bugs (Triatominae)

Brown

Rodríguez‐Ruano

Poosakkannu

et al. 2020

Preprint

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BackgroundKissing bugs (Triatominae) are blood-feeding insects best known as the vectors of Trypanosoma cruzi, the causative agent of Chagas' disease. Considering the high epidemiological relevance of these vectors, their biology and bacterial symbiosis remains surprisingly understudied. While previous investigations revealed generally low individual complexity but high among-individual variability of the triatomine microbiomes, any consistent microbiome determinants have not yet been identified across multiple Triatominae species. Methods To obtain a more comprehensive view of triatomine microbiomes, we investigated the host-microbiome relationship of five Triatoma species sampled from white-throated woodrat (Neotoma albigula) nests in multiple locations across the USA. We applied optimized 16S rRNA gene metabarcoding with a novel 18S rRNA gene blocking primer to a set of 170 T. cruzi negative individuals across all six instars. Results Triatomine gut microbiome composition is strongly influenced by three principal factors: ontogeny, species identity, and the environment. The microbiomes are characterised by significant loss in bacterial diversity throughout ontogenetic development. First instars possess the highest bacterial diversity while adult microbiomes are routinely dominated by a single taxon. Primarily, the bacterial genus Dietzia dominates late-stage nymphs and adults of T. rubida, T. protracta, and T. lecticularia, but is not present in the phylogenetically more distant T. gerstaeckeri and T. sanguisuga. Species-specific microbiome composition, particularly pronounced in early instars, is further modulated by locality-specific effects. In addition, pathogenic bacteria of the genus Bartonella, acquired from the vertebrate hosts, are an abundant component of Triatoma microbiomes. ConclusionOur study is the first to demonstrate deterministic patterns in microbiome composition among all life stages and multiple Triatoma species. We hypothesize that triatomine microbiome assemblages are produced by species-and life stage-dependent uptake of environmental bacteria and multiple indirect transmission strategies that promote bacterial transfer between individuals. Altogether, our study highlights the complexity of Triatominae symbiosis with bacteria and warrant further investigation to understand microbiome function in these important vectors.

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The midgut microbiota plays an essential role in sand fly vector competence forLeishmania major

Cited by 72 publications

References 38 publications

CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

Plant-feeding phlebotomine sand flies, vectors of leishmaniasis, prefer Cannabis sativa

Ontogeny, species identity and environment dominate microbiome dynamics in wild populations of kissing bugs (Triatominae)

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