Project description:The role of domestic and peridomestic animals in vector-borne diseases is very important as they share a common environment with people having the potential to extend the network of pathogen transmission to humans. The most significant vector-borne infectious diseases that are shared by man, domestic and peridomestic animals are leishmaniosis, borreliosis, bartonellosis, ehrlichiosis, rickettsiosis and anaplasmosis with fleas acting as main vectors in the transmission of some of these diseases. Flea-borne diseases which are relevant in Europe include plague (caused by Yersinia pestis), murine typhus (caused by Rickettsia typhi), flea-borne spotted fever (Rickettsia felis), and cat scratch disease (Bartonella henselae). In the present study, mNGS was applied to detect and understand the composition of the microbial communities of five different species of fleas (Archaeopsylla erinacei, Ctenocephalides felis, Spilopsyllus cuniculi, Pulex irritans and Ctenocephalides canis) collected on dogs, cats and hedgehogs from Andalusia (Spain) to know the prevalence of pathogenic bacteria among synanthropic fleas. Based on our results, we could conclude that Pulicidae family encompassed those flea species with a close contact with humans and therefore more involved in the potential transmission of infectious diseases. The higher relative abundance of the Phylum Pseudomanadota was mainly due to the presence of the endosymbiont Wolbachia, as well as to notice a high relative abundance of both genera Rickettsia and Bartonella in all flea species. For the first time, we detected Babesia sp. in all species tested, especially with higher abundance in S. cuniculi collected from cats emphasizing the need for further investigation into its potential implications as vectors. Our results also demonstrate that the microbiota composition of fleas is largely influenced by the host they parasitize. Lastly, statistical analyses of microbiota allowed for the ecological separation of flea species, with individuals from these five species clustering distinctly each other.
Project description:Yersinia pestis, the etiologic agent of plague, emerged as a flea-borne pathogen only within the last 6,000 years. Just five simple genetic changes in the Yersinia pseudotuberculosis progenitor, which served to eliminate toxicity to fleas and to enhance survival and biofilm formation in the flea digestive tract, were key to the transition to the arthropod-borne transmission route. To gain a deeper understanding of the genetic basis for the development of a transmissible biofilm infection in the flea foregut, we evaluated additional gene differences and performed in vivo transcriptional profiling of Y. pestis, Y. pseudotuberculosis wild-type (unable to form biofilm in the flea foregut), and a Y. pseudotuberculosis mutant strain (able to produce foregut-blocking biofilm in fleas) recovered from fleas 1 day and 14 days after an infectious bloodmeal. Surprisingly, the Y. pseudotuberculosis mutations that increased c-di-GMP levels and enabled biofilm development in the flea did not change expression levels of the hms genes responsible for the synthesis and export of the extracellular polysaccharide matrix required for mature biofilm formation. The Y. pseudotuberculosis mutant uniquely expressed much higher levels of one of the Yersinia Type VI secretion systems (T6SS-4) in the flea, and this locus was required for flea blockage by Y. pseudotuberculosis, but not by Y. pestis. Significant differences between the two species in expression of several metabolism genes, the Psa fimbrial genes, quorum sensing related genes, transcriptional regulators, and stress response genes were evident during flea infection. The results provide insights into how Y. pestis has adapted to life in its flea vector and point to evolutionary changes in the regulation of biofilm development pathways in these two closely related species