Molecular detection of Rickettsia felis in fleas and ticks collected from dogs and cats of Puebla, Mexico.

Fleas and ticks represent the two main groups of ectoparasites that infest companion animals. In particular, the flea Ctenocephalides felis felis and several members of the Rhipicephalus sanguineus complex are the main vectors of a wide range of pathogens on the American continent. They are competent vectors for several members of the genus Rickettsia, which encompass at least 15 pathogenic obligate intracellular bacteria that colonize the endothelial cells of vertebrates. In Mexico, 10 species of Rickettsia belonging to three groups have been detected in six species of ectoparasites from dogs in 9 of the 32 states of the country. However, in some larger regions of the country, active epidemiological surveillance has not been carried out. For this reason, the aim of this study was to identify the presence of members of the genus Rickettsia in fleas and ticks of dogs and cats in the state of Puebla, Mexico. A cross-sectional study was carried out to collect ectoparasites of dogs and cats during August to November 2019. Samples were fixed in 70% ethanol and examined to identify the presence of Rickettsia DNA by the amplification and sequencing of specific fragments of the gltA and ompB genes using conventional PCR. The recovered sequences were compared with those deposited in GenBank, and phylogenetic analyses were carried out to identify the position of the pathogens detected with respect to the valid species previously reported worldwide. Additionally, ecological parameters of the ectoparasite infestations were also calculated. We recovered 196 ectoparasites belonging to two species, 33 C. felis felis and 163 R. sanguineus s.l. (Rhipicephalus linnaei), parasitizing 46 hosts (42 dogs and 4 cats) in 11 localities of the state of Puebla. We detected the presence of Rickettsia felis in three pools of C. felis felis, and five from R. sanguineus s.l. Our work provides the first record of R. felis in hard ticks of Mexico and Central America, with new collection localities for this pathogen in central Mexico.

Molecular Evidence for Flea-Borne Rickettsiosis in Febrile Patients from Madagascar.

Rickettsiae may cause febrile infections in humans in tropical and subtropical regions. From Madagascar, no molecular data on the role of rickettsioses in febrile patients are available. Blood samples from patients presenting with fever in the area of the capital Antananarivo were screened for the presence of rickettsial DNA. EDTA (ethylenediaminetetraacetic acid) blood from 1020 patients presenting with pyrexia > 38.5 °C was analyzed by gltA-specific qPCR. Positive samples were confirmed by ompB-specific qPCR. From confirmed samples, the gltA amplicons were sequenced and subjected to phylogenetic analysis. From five gltA-reactive samples, two were confirmed by ompB-specific qPCR. The gltA sequence in the sample taken from a 38-year-old female showed 100% homology with R. typhi. The other sample taken from a 1.5-year-old infant was 100% homologous to R. felis. Tick-borne rickettsiae were not identified. The overall rate of febrile patients with molecular evidence for a rickettsial infection from the Madagascan study site was 0.2% (2/1020 patients). Flea-borne rickettsiosis is a rare but neglected cause of infection in Madagascar. Accurate diagnosis may prompt adequate antimicrobial treatment.

First Detection of Rickettsia typhi and Rickettsia felis in Fleas Collected From Client-Owned Companion Animals in the Southern Great Plains.

Flea-borne rickettsiosis occurs worldwide and includes a number of pathogens, namely, Rickettsia typhi and Rickettsia felis. Most studies in the United States have occurred in southern Texas and California where flea-borne rickettsiosis is endemic, resulting in a lack of information from other regions of the country. Between March and August 2016, 222 fleas were collected from 52 client-owned dogs and cats in two urban areas in Oklahoma. Fleas were identified using morphological characteristics then pooled and tested by polymerase chain reaction (PCR) using published primers for gltA, ompB, and 17-kDa. The majority (98.6%) of fleas collected were Ctenocephalides felis (Bouché) followed by Pulex irritans (L) (1.4%). Overall, fleas collected from 30.0% (6/20) cats and 43.8% (14/32) dogs were infected with R. felis. Three C. felis-pools collected from three dogs, two in the Enid area (central Oklahoma) and one in the Elk City area (western Oklahoma), were infected with R. typhi as well as R. felis. 'Candidatus R. senegalensis' was detected in one pool of fleas taken from a cat in Oklahoma City. This is the first evidence that flea-borne Rickettsia species occur in fleas obtained from client-owned dogs and cats in the Great Plains region. The impact of these Rickettsia species on public health in the region needs further investigation.