Searching for common patterns in parasite ecology: species and host contributions to beta-diversity in helminths of South African ungulates and fleas of South American rodents.

We searched for common patterns in parasite ecology by investigating species and host contributions to the beta-diversity of infracommunities (=assemblages of parasites harboured by a host individual) in helminths of three species of South African ungulates and fleas of 11 species of South American rodents, assuming that a comparison of patterns in distinctly different parasites and hosts would allow us to judge the generality or, at least, commonness of these patterns. We used data on species' composition and numbers of parasites and asked whether (i) parasite species' attributes (life cycle, transmission mode, and host specificity in helminths; possession of sclerotized combs, microhabitat preference, and host specificity in fleas) or their population structure (mean abundance and/or prevalence) and (ii) host characteristics (sex and age) affect parasite and host species' contributions to parasite beta-diversity (SCBD and HCBD, respectively). We found that parasite species' morphological and ecological attributes were mostly not associated with their SCBD. In contrast, parasite SCBD, in both ungulates and rodents, significantly increased with either parasite mean abundance or prevalence or both. The effect of host characteristics on HCBD was detected in a few hosts only. In general, parasite infracommunities' beta-diversity appeared to be driven by variation in parasite species rather than the uniqueness of the assemblages harboured by individual hosts. We conclude that some ecological patterns (such as the relationships between SCBD and parasite abundance/prevalence) appear to be common and do not differ between different host-parasite associations in different geographic regions, whereas other patterns (the relationships between SCBD and parasite species' attributes) are contingent and depend on parasite and host identities.

Fleas (Siphonaptera, Latreille, 1825) from Rio Grande do Sul State, Brazil: Species Diversity, Hosts, and One Health Approach.

Background: Fleas are ectoparasitic insects with holometabolous development. It has a hematophagous habit with mouthparts adapted to sting and suck its hosts. There are about 3000 species in the world, ∼61 in Brazil, and 19 in Rio Grande do Sul state. The objective of the research is to catalog the diversity of fleas recorded in the state, their respective hosts, and endosymbionts. Materials and Methods: To this end, a search was carried out in the scientific literature, from articles, books, to abstracts submitted to congresses. Results: The 19 species of fleas occurring in Rio Grande do Sul are divided into 7 families and 10 genera. These ectoparasites, in addition to being found in the environment, were associated with 10 different families of hosts in Rio Grande do Sul, and on the endosymbiont, agents found associated with fleas, there were 7 different species. The main agents researched in the state are Rickettsia spp. and Bartonella spp. The relationships between parasites, hosts, environment, and etiological agents present different scenarios, whether anthropized or conserved, but unknown. Sometimes, this overlap, a factor that aggravates the possibility of spillovers, either from cosmopolitan fleas in these conserved areas, or from their endosymbionts. Conclusion: Thus, it is important to characterize the environment so that the complexities of each location are known for the adoption of environmental and public health policies in each case. The challenges are extensive, but necessary in view of the One Health perspective.

Tick and flea infestations in captive Tapirus terrestris and Tapirus kabomani (Perissodactyla: Tapiridae) in the Brazilian Amazon.

The present study aimed to identify tick and flea species infesting captive tapirs (Tapirus terrestris and Tapirus kabomani) in the Brazilian Amazon. Ectoparasites were collected from tapirs chemically restrained in Zoo-botanical Parks, breeding facilities, conservationists and, environmental compensation areas in the states of Amapá, Amazonas, and Pará. After collection, the tick and flea specimens were placed in plastic pots containing isopropanol and identified according to dichotomous keys. Ectoparasite infestations were observed in 55% (18/33) tapirs, of which 61% (11/18) were single infestations with ticks or fleas, and 39% (7/18) were mixed infestations with different species of ticks and/or fleas. In total, 227 ticks (15 larvae, 107 nymphs, 38 females and 67 males) and 14 fleas (eight females and six males) were collected, identifying four tick species (Amblyomma cajennense sensu stricto , Amblyomma naponense, Amblyomma oblongoguttatum, and Amblyomma pacae) and three flea species and/or subspecies (Ctenocephalides felis felis, Rhopalopsyllus australis australis, and Tunga penetrans). In conclusion, infestations with Ixodidae ticks and Rhopalopsyllidae, Pulicidae and Tungidae fleas were found in captive tapirs in the Brazilian Amazon, and A. cajennense s.s. was the most frequent species. Additionally, the present study reports new associations between A. pacae and R. a. australis with T. terrestris and between A. cajennense s.s. with T. kabomani.

Host specificity of flea parasites of mammals from the Andean Biogeographic Region.

Host specificity of fleas affects their biodiversity that plays a major role in determining the potential transmission routes by pathogens through vertebrate hosts, including humans. In the Biogeographic Andean region, numerous systematic and ecological studies have been conducted, revealing a high diversity of flea taxa of mammals and the presence of pathogenic organisms transmitted by fleas; however, the degree of preference with which each flea species associates with a mammal host remains poorly understood in this region. Herein, host specificity in mammal fleas from the Andean region was analysed. We employed the number of host species for each flea species and the index of host specificity STD *. Following the literature, 144 species and 13 subspecies of fleas (31 genera and 10 families) have been described in the Andean biogeographic region; 76 taxa are endemic to this region. To carry out the analyses of host specificity, we considered 1759 records of fleas collected from 124 species and 59 genera of wild and domestic mammals, mostly rodent species (85.9%). Our results indicate that typical Andean fleas are genus or family host specific (mostly STD * less than 3.0). More diverse mammal hosts are parasitized by more diverse flea genera and families and these hosts are phylogenetically related. Otherwise, these hosts are associated with different flea lineages, suggesting the interaction of ecological and evolutionary mechanisms (host-switching, ecological adaptations and co-evolutionary alternation). The fields of disease ecology and One Health are considering the host specificity of arthropod vectors as an important point to understand the mechanisms of emergence and re-emergence of diseases. Our results allow us to estimate the risk of diseases involving fleas in the Andean region.

Ecology of fleas and their hosts in the trifinio of north-east Argentina: first detection of Rickettsia asembonensis in Ctenocephalides felis felis in Argentina.

Fleas are important in public health due to their role as parasites and vectors of pathogens, including Rickettsia. The aim of this study was to evaluate the diversity, abundance and prevalence of fleas and the presence of Rickettsia in the trifinio of north-east Argentina. Fleas from household and synanthropic animals were obtained from urban and periurban areas. They were taxonomically identified and samples of 227 fleas in 86 pools were analysed by polymerase chain reaction targeting the gltA and ompB genes of Rickettsia spp. The study revealed that Ctenocephalides felis felis was dominant on dogs, cats and opossums, with higher prevalence in the periurban area. The Shannon-Wiener and Morisita-Horn indices expressed differences in the diversity and similarity values of the absolute abundances of the species between the areas compared. DNA amplifications revealed 30.8% C. f. felis pools positive for Rickettsia spp. Phylogenetic analysis showed that the haplotype obtained was identical to Rickettsia asembonensis from Peru and Brazil. This is the first detection in Argentina of R. asembonensis that infects C. f. felis, and we emphasize the importance of conducting research from a 'One Health' perspective on the role of opossums and rodents in the integration of the transmission cycles of rickettsial bacteria.

MORPHOLOGICAL AND MOLECULAR CHARACTERIZATION OF THE PLAGUE VECTOR XENOPSYLLA BRASILIENSIS.

Members of the flea family Pulicidae have been the focus of many studies due to their significance as diseases vectors of medical and veterinary importance and their cosmopolitan distribution. They often exhibit variation in morphological features that can make correct species identification and management challenging. This may also apply to Xenopsylla brasiliensis (Baker, 1904), an important plague vector. In the current study, we aimed to provide genetic tools for reliable species identification using a DNA barcoding approach. A total of 73 flea specimens was collected from a native host (Namaqua rock mouse, Micaelamys namaquensis) in South Africa and identified morphologically. In addition, we took measurements of 7 morphological characteristics. Subsequently, we successfully generated barcodes of the mitochondrial cytochrome c oxidase subunit I (COI) gene for X. brasiliensis. We validated this approach by comparing our data to COI sequences from Rwandan X. brasiliensis. While sequences from both regions suggested a close relationship between the 2 X. brasiliensis populations, both haplotype and nucleotide diversity were substantially larger for the South African specimens. This may be attributed to human-assisted spread, differences in habitat, and/or host species sampled and merits further study in the future.

Fleas and ticks in armadillos from Argentinean Patagonia: Diversity, abundance and distribution.

In Argentina, quantitative information on the composition and structure of assemblages of arthropod parasites in Xenarthra is scarce. The aim of this study was to describe and compare the community of arthropod parasites of Chaetophractus villosus and Zaedyus pichiy in the Argentinean Patagonia. A total of 1300 ectoparasites (1224 fleas and 76 ticks) were collected from both host species. Seven different species were found, namely Phthiropsylla agenoris, Malacopsylla grossiventris, Hectopsylla broscus, Tunga penetrans, Tunga perforans (Siphonaptera), Amblyomma pseudoconcolor and Amblyomma auricularium s.l. (Ixodida). Four species were present in both host armadillos (P. agenoris, M. grossiventris, H. broscus and A. pseudoconcolor). Phthiropsylla agenoris was the most prevalent and abundant ectoparasite showing significant differences in prevalences, mean intensities and mean abundance. Malacopsylla grossiventris only showed significant differences in mean intensity. The structure and composition of the ectoparasite assemblage and the high values of ectoparasites aggregation recorded in the Patagonian armadillos strongly suggest that these associations are stable throughout their geographical distribution.

Beta and phylogenetic diversities tell complementary stories about ecological networks biogeography.

The beta-diversity of interactions between communities does not necessarily correspond to the differences related to their species composition because interactions show greater variability than species co-occurrence. Additionally, the structure of species interaction networks can itself vary over spatial gradients, thereby adding constraints on the dissimilarity of communities in space. We used published data on the parasitism interaction between fleas and small mammals in 51 regions of the Palearctic to investigate how beta-diversity of networks and phylogenetic diversity are related. The networks could be separated in groups based on the metrics that best described the differences between them, and these groups were also geographically structured. We also found that each network beta-diversity index relates in a particular way with phylogenetically community dissimilarity, reinforcing that some of these indexes have a strong phylogenetic component. Our results clarify important aspects of the biogeography of hosts and parasites communities in Eurasia, while suggesting that networks beta-diversity and phylogenetic dissimilarity interact with the environment in different ways.

Fleas (Siphonaptera) Parasitizing Peridomestic and Indigenous Mammals in Panamá and Screening of Selected Fleas for Vector-Borne Bacterial Pathogens.

In total, 341 fleas belonging to 16 species were collected from 78 host mammals belonging to 10 species in Panamá from 2010 to 2016. The cat flea, Ctenocephalides felis (Bouché) predominated on domestic dogs and was also recorded from domestic cats, the raccoon, Procyon lotor (Linnaeus) and the common opossum, Didelphis marsupialis Linnaeus. The largest number of flea species (7) was recorded from D. marsupialis and the most common flea on that host was the ctenophthalmid, Adoratopsylla intermedia copha Jordan. One Oriental rat flea, Xenopsylla cheopis (Rothschild), was collected from D. marsupialis. Native rodents were parasitized by indigenous ceratophyllid, rhopalopsyllid, and stephanocircid fleas. The Mexican deermouse, Peromyscus mexicanus (Saussure), was parasitized by six species of ceratophyllids belonging to the mostly Central American genera, Baculomeris, Jellisonia, Kohlsia and Plusaetis. The long-tailed singing mouse, Scotinomys xerampelinus (Bangs), was parasitized by Plocopsylla scotinomi Tipton and Méndez, the only species of stephanocircid flea known from Central America. Twenty-six pools of extracted flea DNA representing 5 flea species (C. felis, Pulex echidnophagoides (Wagner), Pulex simulans Baker, A. intermedia copha, and P. scotinomi) and 79 individual fleas were all real-time polymerase chain reaction negative for Rickettsia felis, Rickettsia typhi, and Bartonella henselae.

Detection of Bartonella sp. and a novel spotted fever group Rickettsia sp. in Neotropical fleas of wild rodents (Cricetidae) from Southern Brazil.

The Neotropical region shows a great diversity of fleas, comprising more than 50 genera. The importance of the study of fleas is linked to their potential role as disease vectors. The aim of this study is to investigate the presence of Rickettsia spp. and Bartonella spp. in Neotropical fleas collected from wild rodents in Southern Brazil. From 350 rodents captured, 30 were parasitized by fleas. A total of 61 fleas belonging to two genera and six different species were collected (Craneopsylla minerva minerva, Polygenis occidentalis occidentalis, Polygenis platensis, Polygenis pradoi, Polygenis rimatus, and Polygenis roberti roberti). In 13 % of fleas of three different species (C. minerva, P. platensis, and P. pradoi) Rickettsia sp. DNA was found. Phylogenetic analysis of concatenated sequences of gltA, htrA, and ompA genes showed that Rickettsia sp. found in rodent fleas (referred as strain Taim) grouped together with Spotted Fever Group Rickettsia. In reference to Bartonella spp., five genotypes were identified in seven fleas of two species (C. minerva and P. platensis) and in five rodent spleens. Also, 207 frozen samples of wild rodents were screened for these pathogens: while none was positive for Rickettsia spp.; five rodent spleens were PCR-positive for Bartonella spp.. Herein, we show the detection of potential novel variants of Bartonella sp. and Rickettsia sp. in fleas collected of wild rodents from Southern Brazil. Further studies are needed to fully characterize these microorganisms, as well as to improve the knowledge on the potential role of Neotropical flea species as diseases vectors.

Molecular detection of Rickettsia in fleas from micromammals in Chile.

BACKGROUND: Rickettsial diseases are considered important in public health due to their dispersal capacity determined by the particular characteristics of their reservoirs and/or vectors. Among the latter, fleas play an important role, since the vast majority of species parasitize wild and invasive rodents, so their detection is relevant to be able to monitor potential emerging diseases. The aim of this study was to detect, characterize, and compare Rickettsia spp. from the fleas of micromammals in areas with different human population densities in Chile. METHODS: The presence of Rickettsia spp. was evaluated by standard polymerase chain reaction (PCR) and sequencing in 1315 fleas collected from 1512 micromammals in 29 locations, with different human population densities in Chile. A generalized linear model (GLM) was used to identify the variables that may explain Rickettsia prevalence in fleas. RESULTS: DNA of Rickettsia spp. was identified in 13.2% (174 of 1315) of fleas tested. Fifteen flea species were found to be Rickettsia-positive. The prevalence of Rickettsia spp. was higher in winter, semi-arid region and natural areas, and the infection levels in fleas varied between species of flea. The prevalence of Rickettsia among flea species ranged between 0-35.1%. Areas of lower human density showed the highest prevalence of Rickettsia. The phylogenetic tree showed two well-differentiated clades with Rickettsia bellii positioned as basal in one clade. The second clade was subdivided into two subclades of species related to Rickettsia of the spotted fever group. CONCLUSIONS: To our knowledge, this is the first report of the occurrence and molecular characterization of Rickettsia spp. in 15 flea species of micromammals in Chile. In this study, fleas were detected carrying Rickettsia DNA with zoonotic potential, mainly in villages and natural areas of Chile. Considering that there are differences in the prevalence of Rickettsia in fleas associated with different factors, more investigations are needed to further understand the ecology of Rickettsia in fleas and their implications for human health.

Molecular detection and identification of Rickettsia felis in Polygenis (Siphonaptera, Rhopalopsyllidae, Rhopalopsyllinae) associated with cricetid rodents in a rural area from central Argentina.

The aim of this study was to detect and identify the presence of Rickettsia in fleas associated with cricetid rodents from northeastern Buenos Aires Province, Argentina. Sixteen fleas belonging to three species of Polygenis were collected from 56 cricetid rodents and analyzed for the presence of Rickettsia performing the conventional polymerase chain reaction (PCR) technique. Only one specimen of Polygenis (Polygenis) axius axius collected from Oxymycterus rufus was positive for Rickettsia felis using the gltA gene, and to ompA gene. This is the first report of R. felis in a Rhopalopsyllidae flea from Argentina, and the first detection of this bacterium in P. (P.) a. axius. Since both, O. rufus and P. (P.) a. axius, are common in areas close to humans, and enzootic cycle of R. felis is not fully understood, the results herein obtained might be of epidemiological importance. Further studies are needed in order to analyze the capacity of the species of Polygenis to transmit R. felis.

Rickettsia asembonensis: New records associated with the cat flea (Ctenocephalides felis felis) in Mexico.

In the present work, we report for the first time the presence of an emerging rickettsial agent in Mexico, Rickettsia asembonensis, recorded in the cat flea (Ctenocephalides felis felis) collected in dogs from the states of Morelos and Veracruz. Possible implications for public health and its contribution to the biodiversity of the country are discussed.

Does bathing affect tick and flea burdens and ectoparasiticide effectiveness of a spot-on formulation (fipronil + (S)-methoprene) for dogs?

This work evaluated tick and flea burdens and the efficacy of a single topical application of 10% fipronil + 9% (S)-methoprene spot-on against experimental infestations of Rhipicephalus sanguineus sensu lato and Ctenocephalides felis felis on dogs that were submitted to regular baths for 56-days post-treatment. Four treatments (n = 24) were evaluated: no chemical treatment and no bathing (T01); no chemical treatment + bathing (T02); chemical treatment and no bathing (T03) and chemical treatment + bathing (T04). Dogs were infested with adult ticks and fleas seven days pre-treatment (-7). The ectoparasites were then counted and removed on day -5, followed by a new infestation on day -2 and treatment on day 0. Ticks and fleas were then counted without removal on day 1 and counted with removal on day 2. The dogs were then repeatedly infested with both ectoparasites on days 5, 12, 19, 26, 33, 40, 47 and 54 post-treatment. Baths and ectoparasite counts with removal were performed every 48 h before and after experimental infestation, respectively. Animals of T04 had lower (P ≤ 0.05) tick burden than animals from T01 and T02 from day 2 to day 28. For fleas, the counts varied among days of the study. Dogs from T03 presented lower (P ≤ 0.05) flea burden than dogs from T01 from day 1 until day 56. The same was true for T04 when compared to T01 from day 1 to day 42. Animals of T02 greater (P ≤ 0.05) flea burden than animals of T04 in all dates post treatment. In addition, dogs from T02 presented higher (P ≤ 0.05) flea burden than T01 from day 7 to day 28 and from day 49 to day 56. In sum, the efficacy of the spot-on formulation used declined over the time against both ectoparasites. The efficacy for ticks ranged from 0.0 to 96.5% and 0.0 to 98.52% for T03 and T04, respectively. While for fleas, the efficacy of the formulation ranged from 67.41 to 100% for T03 and 40.18 to 100% for T04. So, bathing dogs that were not treated increased C. f. felis burden but not R. sanguineus s. l. burden. The residual efficacy of the product had a shorter duration against these ectoparasites for dogs that received subsequent bathing.

Ectoparasites of Didelphis virginiana From Yucatan, Mexico.

A faunal study is presented to determine the species of ectoparasites found in the opossum Didelphis virginiana (Kerr) (Marsupialia: Didelphidae). For this, Tomahawk traps were placed in the peridomiciles of a rural town in Yucatán, to capture individuals of this marsupial and proceed to the collection of their ectoparasites. A total of 3,023 arthropods were collected from 145 opossums. The most frequent ectoparasites were the acarines Ornithodoros (Alectorobius) nr. talaje (64.8%) (Argasidae); Ornithonyssus wernecki Fonseca (53.8%) (Macronyssidae) and Didelphilicus serrifer Fain (25.5%) (Atopomelidae); the ticks Amblyomma parvum Aragão (8.3%) and A. mixtum Koch (10.3%) (Ixodidae); and the fleas Ctenocephalides felis (Bouché) (20.7%) and Pulex simulans Baker (8.3%) (Pulicidae). It is concluded that the ectoparasite fauna of this marsupial is mainly composed of euryxenous organisms, which have been documented as vectors of diseases caused by rickettsial bacteria. Since D. virginiana is widely adapted to the peridomiciliary environment in the region, this study allows recognizing those that represent a potential risk for the transmission of vector-borne zoonotic diseases.

First report of Siphonaptera parasites in Canis latrans in the Flora and Fauna Protection Area, Médanos de Samalayuca Chihuahua, Mexico.

Siphonaptera are hematophage parasite vectors of both human and animal diseases. We aimed to identify ectoparasites parasitizing a coyote population (Canis latrans) in the northwest region of the Flora and Fauna Protection Area Médanos de Samalayuca, Chihuahua, Mexico. We captured 21 coyotes (15 males and 6 females) during the summer and winter of 2018. The individuals were anesthetized and thoroughly examined for ectoparasites. We found that 43% of the coyotes were infested. Based on characteristics such as the absence of pronotal and genal combs in the head, we identified 15 specimens as Pulex irritans. This is the first report of P. irritans in coyotes in Médanos de Samalayuca Chihuahua, Mexico.

Molecular detection and identification of Bartonella in the cat flea Ctenocephalides felis felis collected from companion animals in a border area in northeastern Argentina.

Molecular methods were used to detect and identify Bartonella species in the cat fleas Ctenocephalides felis felis from Puerto Iguazú, a border area in northeastern Argentina. The fleas were collected from 12 household animals, 9 dogs (Canis lupus familiaris) and 3 cats (Felis silvestris catus) during July 2016. Out of 15C. f. felis analyzed for PCR, only one flea collected from a cat was positive (6.66%) in screened for Bartonella spp. based on the gltA gene. Bartonella clarridgeiae was identified in the genetic analyses, this specimen clustered monophyletically with others B. clarridgeiae isolated from different geographical origins (1.0 PP), even, all shared the same haplotype. The results obtained provide evidence of the presence of B. clarridgeiae in cat fleas from Argentina suggesting the probable presence of related flea-borne diseases in the region and the role of cat fleas in the transmission of Bartonella among mammals including humans.

Candidatus Rickettsia senegalensis in Cat Fleas (Siphonaptera: Pulicidae) Collected from Dogs and Cats in Cauca, Colombia.

Rickettsia typhi and Rickettsia felis (Rickettsiales: Rickettsiaceae) are flea-transmitted pathogens. They are important causes of acute febrile illness throughout the world. We, therefore, sought to identify the rickettsial species present in the fleas of dogs and cats in the department of Cauca, Colombia. In this study, we collected 1,242 fleas from 132 dogs and 43 fleas from 11 cats. All fleas were morphologically identified as Ctenocephalides felis (Bouché) adults and organized in pools for DNA extraction (234 pools from dogs and 11 from cats). The gltA gene from rickettsiae was targeted for screening amplification using conventional PCR. In total, 144 of the 245 pools (58.7%) were positive. The positive samples were then processed for the amplification of the 17kDa antigen gene (144/144; 100% positive) and sca5 gene (140/144; 97.2% positive). In addition, restriction enzyme length polymorphism analysis using NlaIV on the amplified product of the sca5 gene demonstrated several organisms: 21/140 (15%) were R. felis, 118/140 (84.3%) were Rickettsia asemboensis, and 1/140 (0.7%) were Candidatus Rickettsia senegalensis. Subsequent sequencing confirmed Candidatus Rickettsia senegalensis in C. felis collected from dogs the first reported from Colombia.

Molecular detection of Bartonella in fleas (Hexapoda, Siphonaptera) collected from wild rodents (Cricetidae, Sigmodontinae) from Argentina.

Bartonella are facultative intracellular Gram-negative bacteria, transmitted mainly by hematophagous arthropods, and the rodents act as a natural reservoir. Different species of Bartonella associated with rodents have been implicated as causing human disease. Studies from Argentina are scarce and no Bartonella from fleas have been reported previously. The present study investigated the presence of Bartonella spp. in fleas associated with sigmodontine rodents in four localities of the Santa Cruz Province, Argentina. In total, 51 fleas (four species) were analysed of which 41.2% were found to be positive for the gltA gene fragment via a nested polymerase chain reaction. All positive fleas were of the species Neotyphloceras crackensis from three different localities. Eight of the 21 amplified samples were sequenced, and the presence of three different genotypes was detected with an identity of 95.5-98.8% amongst themselves. Bartonella genotypes from American rodents and rodent fleas were recovered in a monophyletic group. Similarly, most of the Peruvian and all Argentinean variants constitute a natural group sister of the American remainder. The importance of the Bartonella spp. with respect to public health is unknown, although future studies could provide evidence of the possible involvement of N. crackensis in the Bartonella transmission cycles.

Detection of Rickettsia spp. and Bartonella spp. in Ctenocephalides felis fleas from free-ranging crab-eating foxes (Cerdocyon thous).

Fleas are insects with a worldwide distribution that have been implicated in the transmission of several pathogens. The present study aimed to investigate the presence of Rickettsia spp. (Rickettsiales: Rickettsiaceae) and Bartonella spp. (Rhizobiales: Bartonellaceae) in fleas from free-ranging crab-eating foxes Cerdocyon thous (Linnaeus, 1766) (Carnivora: Canidae) from Rio Grande do Sul, southern Brazil. Fleas were collected manually from animals and used for the molecular detection of Rickettsia spp. and Bartonella spp. Twenty-nine C. thous were sampled in six municipalities. Four foxes were parasitized by 10 fleas, all of which were identified as Ctenocephalides felis (Bouché, 1935) (Siphonaptera: Pulicidae). DNA from Rickettsia felis Bouyer et al., 2001 and Rickettsia asembonensis Maina et al., 2016 were found in three and eight fleas, respectively. In four fleas, DNA of Bartonella sp. was identified. Phylogenetic analysis grouped Bartonella sp. together with other genotypes previously reported in C. felis worldwide. The scenario described in the present study highlights a Neotropical canid parasitized by the invasive cosmopolitan cat flea, which in turn, is carrying potentially invasive vector-borne microorganisms. These findings suggest that C. felis is adapted to wild hosts in wilderness areas in southern Brazil, hypothetically exposing the Neotropical fauna to unknown ecological and health disturbances.