Metagenomic analysis of human-biting cat fleas in urban northeastern United States of America reveals an emerging zoonotic pathogen

Ferreira, Francisco C.; Fonseca, Dina M.; Hamilton, George C.; Price, Dana C.

doi:10.1038/s41598-020-72956-x

Cited by 10 publications

(12 citation statements)

References 60 publications

(63 reference statements)

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“…Although not the first study to report both flea genotype and pathogen carriage status [ 2 , 20 ], this study reports a large sample of C. felis genotype and Bartonella, Rickettsia , and Wolbachia infection status. Our data provide evidence that C. felis genotypic haplotype is most likely to be related to the infecting Bartonella and Wolbachia strain but less likely to be related to Rickettsia strain.…”

Section: Discussionmentioning

confidence: 99%

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Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny

et al. 2022

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Background Ctenocephalides felis, the cat flea, is the most common ectoparasite of cats and dogs worldwide. As a cause of flea allergy dermatitis and a vector for two genera of zoonotic pathogens (Bartonella and Rickettsia spp.), the effect of the C. felis microbiome on pathogen transmission and vector survival is of substantial medical importance to both human and veterinary medicine. The aim of this study was to assay the pathogenic and commensal eubacterial microbial communities of individual C. felis from multiple geographic locations and analyze these findings by location, qPCR pathogen prevalence, and flea genetic diversity. Methods 16S Next Generation Sequencing (NGS) was utilized to sequence the microbiome of fleas collected from free-roaming cats, and the cox1 gene was used for flea phylogenetic analysis. NGS data were analyzed for 168 individual fleas from seven locations within the US and UK. Given inconsistency in the genera historically reported to constitute the C. felis microbiome, we utilized the decontam prevalence method followed by literature review to separate contaminants from true microbiome members. Results NGS identified a single dominant and cosmopolitan amplicon sequence variant (ASV) from Rickettsia and Wolbachia while identifying one dominant Bartonella clarridgeiae and one dominant Bartonella henselae/Bartonella koehlerae ASV. Multiple less common ASVs from these genera were detected within restricted geographical ranges. Co-detection of two or more genera (Bartonella, Rickettsia, and/or Wolbachia) or multiple ASVs from a single genus in a single flea was common. Achromobacter, Peptoniphilus, and Rhodococcus were identified as additional candidate members of the C. felis microbiome on the basis of decontam analysis and literature review. Ctenocephalides felis phylogenetic diversity as assessed by the cox1 gene fell within currently characterized clades while identifying seven novel haplotypes. NGS sensitivity and specificity for Bartonella and Rickettsia spp. DNA detection were compared to targeted qPCR. Conclusions Our findings confirm the widespread coinfection of fleas with multiple bacterial genera and strains, proposing three additional microbiome members. The presence of minor Bartonella, Rickettsia, and Wolbachia ASVs was found to vary by location and flea haplotype. These findings have important implications for flea-borne pathogen transmission and control. Graphical Abstract

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Section: Discussionmentioning

confidence: 99%

“…The diversity of C. felis-associated Rickettsia spp. has only recently been appreciated; therefore, little is known regarding their geographic distribution [17][18][19][20]. In addition to pathogenic Bartonella and Rickettsia spp., Wolbachia is widely accepted as a C. felis microbial constituent.…”

Section: Introductionmentioning

confidence: 99%

Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny

et al. 2022

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“…Comparable to tick-borne epidemiology, the recent discovery of R. felis genotypic variants, collectively referred to as R. felis-like organisms (RFLOs), in various arthropods throughout the world has complicated the understanding of flea-borne rickettsioses [89]. Although once presumed as strict endosymbionts, RFLOs have been molecularly detected among vertebrate hosts [93]. Furthermore, cat fleas constitutively infected with R. felis experimentally acquire R. typhi at a lower frequency after subsequent exposure compared to uninfected cat fleas [94].…”

Section: Microbial Interactions and Vector Biologymentioning

confidence: 99%

Unpacking the intricacies of Rickettsia–vector interactions

Laukaitis

Macaluso

2021

Trends in Parasitology

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“…Spotted fever group (SFG) rickettsioses are important neglected zoonoses throughout the world, with expanding known distribution. Also, SFG rickettsiae are transmitted by ticks while lice and fleas are vectors for typhus group rickettsiae [1][2][3][4]. Currently, the reported SFG rickettsioses cases in China are Rickettsia heilongjiangensis, Rickettsia raoultii, Rickettsia slovaca, Rickettsia sibirica and Rickettsia massiliae [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Visual closed dumbbell-mediated isothermal amplification (CDA) for on-site detection of Rickettsia raoultii

Gong

Cai

et al. 2022

PLoS Negl Trop Dis

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Spotted fever group (SFG) rickettsioses are important zoonoses, threatening human health seriously and gradually attracting more attention in the world. SFG rickettsiae are classified as neglected pathogens. If these pathogens are detected at all, they are usually recognized very late in the infection through indirect detection of specific antibodies. Previous studies have shown that Rickettsia raoultii (R. raoultii), a member of the SFG rickettsiae, occurs with increasing incidence in remote countries. Therefore, a rapid detection method for R. raoultii is in urgently need. In this study, a R. raoultii diagnosis method by closed dumbbell-mediated isothermal amplification (R-CDA) assay targeting a conserved sequence of the outer membrane protein A (OmpA) gene with high sensitivity and specificity was developed. This assay offered a rapid and simple method for on-site detection of R. raoultii. Firstly, four pairs of R-CDA primers were designed and the optimum primer set was selected to amplify target gene specifically and effectively. Then, a pair of outer primer was designed to accelerate the reaction based on the inner primers to establish the RO-CDA reaction. In addition, the results of real-time amplification curves, melting curves and end-point colorimetric judgements showed that the established visual RO-CDA reaction could accurately detect R. raoultii without cross-reaction with other closely related pathogens. Furthermore, the detection limit of visual RO-CDA assay was 10 copies/μL, which was feasible for on-site detection with merits of easy-operation, rapidity, high sensitivity, and specificity. In conclusion, the developed RO-CDA detection method could be helpful for pathogen screening and epidemic prevention at the point of care.

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Metagenomic analysis of human-biting cat fleas in urban northeastern United States of America reveals an emerging zoonotic pathogen

Cited by 10 publications

References 60 publications

Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny

Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny

Unpacking the intricacies of Rickettsia–vector interactions

Visual closed dumbbell-mediated isothermal amplification (CDA) for on-site detection of Rickettsia raoultii

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