Detection of Novel Bartonella Strains and Yersinia pestis in Prairie Dogs and Their Fleas (Siphonaptera

Ceratophyllidae and Pulicidae) Using Multiplex Polymerase Chain Reaction

Heather Stevenson-Lerner, Ying Bai, Michael Y. Kosoy, John A. Montenieri, Jennifer L. Lowell, May C. Chu, Kenneth L. Gage

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

We developed a multiplex polymerase chain reaction (PCR) assay that simultaneously detects three types of flea-associated microorganisms. Targets for the assay were sequences encoding portions of the gltA, a 17-kDa antigen, and pla genes of Bartonella spp. Strong et al., Rickettsia spp. da Rocha-Lima, and Yersinia pestis Yersin, respectively. A total of 260 flea samples containing bloodmeal remnants were analyzed from fleas collected from abandoned prairie dog (Cynomys ludovicianus) burrows at the site of an active plague epizootic in Jefferson County, CO. Results indicated that 34 (13.1%) fleas were positive for Bartonella spp., 0 (0%) were positive for Rickettsia spp., and 120 (46.2%) were positive for Y. pestis. Twenty-three (8.8%) of these fleas were coinfected with Bartonella spp. and Y. pestis. A second group of 295 bloodmeal-containing fleas was collected and analyzed from abandoned burrows in Logan County, CO, where a prairie dog die-off had occurred 2-4 mo before the time of sampling. Of these 295 fleas, 7 (2.4%) were positive for Bartonella spp., 0 (0%) were positive for Rickettsia spp., and 46 (15.6%) were positive for Y. pestis. Coinfections were not observed in fleas from the Logan County epizootic site. The multiplex PCR also was used to identify Y pestis and Bartonella in prairie dog blood and tissues. This report represents the first identification of Bartonella from prairie dogs and their fleas. Prairie dog fleas were tested with PCR, and the Bartonella PCR amplicons produced were sequenced and found to be closely related to similar sequences amplified from Bartonella that had been isolated from prairie dog blood samples. Phylogenetic analyses indicate that the sequences of bartonellae from prairie dogs and prairie dog fleas cluster tightly within a clade that is distinct from those containing other known Bartonella genotypes.

Original languageEnglish (US)
Pages (from-to)329-337
Number of pages9
JournalJournal of Medical Entomology
Volume40
Issue number3
StatePublished - May 2003
Externally publishedYes

Fingerprint

Ceratophyllidae
Bartonella
Pulicidae
Yersinia pestis
Siphonaptera
Cynomys
Sciuridae
Multiplex Polymerase Chain Reaction
polymerase chain reaction
Rickettsia
Ctenocephalides canis
burrows
Carbon Monoxide
Cynomys ludovicianus
die-off
plague
blood
assays
Polymerase Chain Reaction
mixed infection

Keywords

  • Bartonella
  • Black-tailed prairie dog
  • Fleas
  • Multiplex polymerase chain reaction
  • Oropsylla
  • Plague
  • Rickettsia
  • Yersinia pestis

ASJC Scopus subject areas

  • Insect Science
  • veterinary(all)

Cite this

Detection of Novel Bartonella Strains and Yersinia pestis in Prairie Dogs and Their Fleas (Siphonaptera : Ceratophyllidae and Pulicidae) Using Multiplex Polymerase Chain Reaction. / Stevenson-Lerner, Heather; Bai, Ying; Kosoy, Michael Y.; Montenieri, John A.; Lowell, Jennifer L.; Chu, May C.; Gage, Kenneth L.

In: Journal of Medical Entomology, Vol. 40, No. 3, 05.2003, p. 329-337.

Research output: Contribution to journalArticle

Stevenson-Lerner, Heather ; Bai, Ying ; Kosoy, Michael Y. ; Montenieri, John A. ; Lowell, Jennifer L. ; Chu, May C. ; Gage, Kenneth L. / Detection of Novel Bartonella Strains and Yersinia pestis in Prairie Dogs and Their Fleas (Siphonaptera : Ceratophyllidae and Pulicidae) Using Multiplex Polymerase Chain Reaction. In: Journal of Medical Entomology. 2003 ; Vol. 40, No. 3. pp. 329-337.
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abstract = "We developed a multiplex polymerase chain reaction (PCR) assay that simultaneously detects three types of flea-associated microorganisms. Targets for the assay were sequences encoding portions of the gltA, a 17-kDa antigen, and pla genes of Bartonella spp. Strong et al., Rickettsia spp. da Rocha-Lima, and Yersinia pestis Yersin, respectively. A total of 260 flea samples containing bloodmeal remnants were analyzed from fleas collected from abandoned prairie dog (Cynomys ludovicianus) burrows at the site of an active plague epizootic in Jefferson County, CO. Results indicated that 34 (13.1{\%}) fleas were positive for Bartonella spp., 0 (0{\%}) were positive for Rickettsia spp., and 120 (46.2{\%}) were positive for Y. pestis. Twenty-three (8.8{\%}) of these fleas were coinfected with Bartonella spp. and Y. pestis. A second group of 295 bloodmeal-containing fleas was collected and analyzed from abandoned burrows in Logan County, CO, where a prairie dog die-off had occurred 2-4 mo before the time of sampling. Of these 295 fleas, 7 (2.4{\%}) were positive for Bartonella spp., 0 (0{\%}) were positive for Rickettsia spp., and 46 (15.6{\%}) were positive for Y. pestis. Coinfections were not observed in fleas from the Logan County epizootic site. The multiplex PCR also was used to identify Y pestis and Bartonella in prairie dog blood and tissues. This report represents the first identification of Bartonella from prairie dogs and their fleas. Prairie dog fleas were tested with PCR, and the Bartonella PCR amplicons produced were sequenced and found to be closely related to similar sequences amplified from Bartonella that had been isolated from prairie dog blood samples. Phylogenetic analyses indicate that the sequences of bartonellae from prairie dogs and prairie dog fleas cluster tightly within a clade that is distinct from those containing other known Bartonella genotypes.",
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