Phylogenetic Evidence for the Existence of Multiple Strains of Rickettsia parkeri in the New World

Free PMC Article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881050/The bacterium Rickettsia parkeri has been reported to infect ticks of the "Amblyomma maculatum species complex" in the New World, where it causes spotted fever illness in humans. In South America, three additional rickettsial strains, namely, Atlantic rainforest, NOD, and Parvitarsum, have been isolated from the ticks Amblyomma ovale, Amblyomma nodosum, and Amblyomma parvitarsum, respectively. These three strains are phylogenetically closely related to R. parkeri, Rickettsia africae, and Rickettsia sibirica Herein, we performed a robust phylogenetic analysis encompassing 5 genes (gltA, ompA, virB4, dnaA, and dnaK) and 3 intergenic spacers (mppE-pur, rrl-rrf-ITS, and rpmE-tRNAfMet) from 41 rickettsial isolates, including different isolates of R. parkeri, R. africae, R. sibirica, Rickettsia conorii, and strains Atlantic rainforest, NOD, and Parvitarsum. In our phylogenetic analyses, all New World isolates grouped in a major clade distinct from the Old World Rickettsia species (R. conorii, R. sibirica, and R. africae). This New World clade was subdivided into the following 4 clades: the R. parkerisensu stricto clade, comprising the type strain Maculatum 20 and all other isolates of R. parkeri from North and South America, associated with ticks of the A. maculatum species complex; the strain NOD clade, comprising two South American isolates from A. nodosum ticks; the Parvitarsum clade, comprising two South American isolates from A. parvitarsum ticks; and the strain Atlantic rainforest clade, comprising six South American isolates from the A. ovale species complex (A. ovale or Amblyomma aureolatum). Under such evidences, we propose that strains Atlantic rainforest, NOD, and Parvitarsum are South American strains of R. parkeriIMPORTANCE Since the description of Rickettsia parkeri infecting ticks of the "Amblyomma maculatum species complex" and humans in the New World, three novel phylogenetic close-related rickettsial isolates were reported in South America. Herein, we provide genetic evidence that these novel isolates, namely, strains Atlantic rainforest, NOD, and Parvitarsum, are South American strains of R. parkeri. Interestingly, each of these R. parkeri strains seems to be primarily associated with a tick species group, namely, R. parkerisensu stricto with the "Amblyomma maculatum species group," R. parkeri strain NOD with Amblyomma nodosum, R. parkeri strain Parvitarsum with Amblyomma parvitarsum, and R. parkeri strain Atlantic rainforest with the "Amblyomma ovale species group." Such rickettsial strain-tick species specificity suggests a coevolution of each tick-strain association. Finally, because R. parkerisensu stricto and R. parkeri strain Atlantic rainforest are human pathogens, the potential of R. parkeri strains NOD and Parvitarsum to be human pathogens cannot be discarded.This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (grant 2011/51979-1 to F.A.N.-B.) and by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior CAPES/PROEX 1841/2016.info:eu-repo/semantics/publishedVersio

Update on tick-borne rickettsioses around the world : a geographic approach [plus erratum]

Tick-borne rickettsioses are caused by obligate intracellular bacteria belonging to the spotted fever group of the genus Rickettsia. These zoonoses are among the oldest known vector-borne diseases. However, in the past 25 years, the scope and importance of the recognized tick-associated rickettsial pathogens have increased dramatically, making this complex of diseases an ideal paradigm for the understanding of emerging and reemerging infections. Several species of tick-borne rickettsiae that were considered nonpathogenic for decades are now associated with human infections, and novel Rickettsia species of undetermined pathogenicity continue to be detected in or isolated from ticks around the world. This remarkable expansion of information has been driven largely by the use of molecular techniques that have facilitated the identification of novel and previously recognized rickettsiae in ticks. New approaches, such as swabbing of eschars to obtain material to be tested by PCR, have emerged in recent years and have played a role in describing emerging tick-borne rickettsioses. Here, we present the current knowledge on tick-borne rickettsiae and rickettsioses using a geographic approach toward the epidemiology of these diseases

Diagnóstico sorológico de erliquiose canina com antígeno brasileiro de Ehrlichia canis Serological diagnosis of canine monocytic ehrlichiosis with Brazilian antigen of Ehrlichia canis

O presente trabalho relata o isolamento de Ehrlichia canis em cultivo de células DH82 e posterior padronização da Reação de Imunofluorescência Indireta (RIFI). Leucócitos de uma cadela experimentalmente infectada com o isolado Jaboticabal de E. canis foram inoculados em cultivo de células DH82. A inoculação foi monitorada após a segunda semana, a cada 5-6 dias, através de exames citológicos e pela amplificação de um fragmento do gene dsb de Ehrlichia pela Reação em Cadeia pela Polimerase (PCR) para confirmação da infecção. A cultura apresentou-se positiva aos 27 dias pós-inoculação pela PCR e aos 28 dias pela citologia. No 33o dia pós-inoculação, observou-se 20% de células infectadas e, aos 53 dias, 60% de infecção. Atualmente, o isolado encontra-se estabelecido em células DH82, com várias passagens atingindo 90-100% de células infectadas entre 7-10 dias após a inoculação. Após o seqüenciamento do produto de PCR, o isolado apresentou-se 100% similar à seqüência correspondente de E. canis depositada no GenBank. As células infectadas foram utilizadas como antígeno para a padronização da RIFI para detecção da infecção em cães.<br>The present study describes a successful isolation of Ehrlichia canis and its establishment in DH82 cells, followed by the development of an Indirect Fluorescent Antibodies Test (IFAT). Leukocytes collected from an experimentally infected dog with the Jaboticabal strain of E. canis were used to inoculate a DH82 cell monolayer. Two weeks later, the inoculated culture was checked for infectivity, every 5-6 days by both cytological staining and PCR, targeting a fragment of the dsb gene. The cell culture showed to be infected by Ehrlichia on day 27 by PCR and on day 28 by cytological staining. By the day 33, the infection rate reached 20% and on day 53, 60%. Currently, the isolate is established in DH82 cells, with several passages reaching 90-100% of infected cells, within 7 to 10 days post inoculation. After sequencing, the amplicon was identical to other E. canis corresponding sequences available in the GenBank. DH82 infected cells were used to standardize an IFAT for the diagnosis of canine ehrlichiosis