Detection of Rickettsia felis in a New World flea species, Anomiopsyllus nudata (Siphonaptera: Ctenophthalmidae)

Heather Stevenson-Lerner, Marcelo B. Labruna, John A. Montenieri, Michael Y. Kosoy, Kenneth L. Gage, David Walker

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The flea and rodent samples studied in this project were collected from field study sites in New Mexico from winter 1998 to spring 2001. During this period, 155 small rodents (14 different species) were live-trapped and combed for the presence of fleas. A total of 253 fleas were collected, comprising 21 species. Two of the 253 fleas collected were polymerase chain reaction (PCR) positive for the Rickettsia 17-kDa protein gene. These two fleas were both Anomiopsyllus nudata Baker, each collected from an individual Neofoma albigula Hartley, on two occasions. Individual fleas positive for the Rickettsia 17-kDa protein gene were then tested with primers targeting the rickettsial genes for citrate synthase (gltA) and two major outer membrane proteins (ompA and ompB). The nucleotide sequences of the PCR products of these two fleas were identical to each other and were 100% (394/394), 100% (1150/1150), 99.8% (469/470), and 99.3% (818/824) similar to the corresponding sequences of the 17-kDa, gltA, ompA, and ompB genes of Rickettsia felis, respectively. Flea homogenates of individual PCR-positive fleas were inoculated into shell vials seeded with Vero cells, and the Giménez stain technique was used to demonstrate the presence of Rickettsia-like organisms in detached cells found in aspirated medium 19 d after inoculation. These cells were harvested and tested by PCR, targeting portions of the 17-kDa and gltA genes, resulting in products 100% identical to R. felis. This work comprises the first report of R. felis detection in a flea species (A. nudata) endemic to the New World.

Original languageEnglish (US)
Pages (from-to)163-167
Number of pages5
JournalJournal of Medical Entomology
Volume42
Issue number2
StatePublished - Mar 2005

Fingerprint

Rickettsia felis
Siphonaptera
Rickettsia
polymerase chain reaction
Polymerase Chain Reaction
genes
Rodentia
rodents
rickettsia-like organisms
Citrate (si)-Synthase
citrate (si)-synthase
Vero Cells
Gene Targeting
outer membrane proteins
cells
Genes

Keywords

  • Anomiopsyllus nudata
  • Flea
  • Neotoma albigula
  • New World
  • Rickettsia felis

ASJC Scopus subject areas

  • Insect Science
  • veterinary(all)

Cite this

Detection of Rickettsia felis in a New World flea species, Anomiopsyllus nudata (Siphonaptera : Ctenophthalmidae). / Stevenson-Lerner, Heather; Labruna, Marcelo B.; Montenieri, John A.; Kosoy, Michael Y.; Gage, Kenneth L.; Walker, David.

In: Journal of Medical Entomology, Vol. 42, No. 2, 03.2005, p. 163-167.

Research output: Contribution to journalArticle

Stevenson-Lerner, Heather ; Labruna, Marcelo B. ; Montenieri, John A. ; Kosoy, Michael Y. ; Gage, Kenneth L. ; Walker, David. / Detection of Rickettsia felis in a New World flea species, Anomiopsyllus nudata (Siphonaptera : Ctenophthalmidae). In: Journal of Medical Entomology. 2005 ; Vol. 42, No. 2. pp. 163-167.
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AB - The flea and rodent samples studied in this project were collected from field study sites in New Mexico from winter 1998 to spring 2001. During this period, 155 small rodents (14 different species) were live-trapped and combed for the presence of fleas. A total of 253 fleas were collected, comprising 21 species. Two of the 253 fleas collected were polymerase chain reaction (PCR) positive for the Rickettsia 17-kDa protein gene. These two fleas were both Anomiopsyllus nudata Baker, each collected from an individual Neofoma albigula Hartley, on two occasions. Individual fleas positive for the Rickettsia 17-kDa protein gene were then tested with primers targeting the rickettsial genes for citrate synthase (gltA) and two major outer membrane proteins (ompA and ompB). The nucleotide sequences of the PCR products of these two fleas were identical to each other and were 100% (394/394), 100% (1150/1150), 99.8% (469/470), and 99.3% (818/824) similar to the corresponding sequences of the 17-kDa, gltA, ompA, and ompB genes of Rickettsia felis, respectively. Flea homogenates of individual PCR-positive fleas were inoculated into shell vials seeded with Vero cells, and the Giménez stain technique was used to demonstrate the presence of Rickettsia-like organisms in detached cells found in aspirated medium 19 d after inoculation. These cells were harvested and tested by PCR, targeting portions of the 17-kDa and gltA genes, resulting in products 100% identical to R. felis. This work comprises the first report of R. felis detection in a flea species (A. nudata) endemic to the New World.

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