Isolation and establishment of the raccoon Ehrlichia-like agent in tick cell culture

Ulrike G Munderloh, Michael J. Yabsley, Staci M. Murphy, M. Page Luttrell, Elizabeth W. Howerth

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Feral animals are reservoirs of emerging human pathogens, as well as carriers of closely related wildlife diseases. The latter may interfere with epidemiologic studies by inducing cross-reactive antibodies, or by providing false positive signals in PCR based tests. We cultured a novel intracellular bacterium from the blood of two raccoons (Procyon lotor): RAC413 and RAC414. RAC413 had been experimentally inoculated with blood from a wild-caught raccoon, and provided the material for a blood passage into RAC414. The microbes grew in Ixodes scapularis (black-legged tick) cells, line ISE6, inoculated either with the leukocyte or erythrocyte fraction of anticoagulated blood. Giemsa-stained cells sampled two and three months after initial inoculation of the cultures revealed inclusions similar to those of Ehrlichia sp., except that individual bacteria commonly were elongated and clustered within endosomes. Electronmicroscopy confirmed the presence of irregularly shaped bacteria with evenly granular bacterioplasm bounded by a unit membrane. 16S rDNA sequencing identified the microbes as the raccoon Ehrlichia-like agent previously detected in feral raccoons from Georgia, United States. In conclusion, the availability of a culture isolate of this agent will facilitate future studies to determine its biology, epidemiologic significance, vector association, and host range. The Ehrlichia-like agent infecting raccoons joins a growing list of tick-borne agents cultivable in tick cells.

Original languageEnglish (US)
Pages (from-to)418-425
Number of pages8
JournalVector-Borne and Zoonotic Diseases
Volume7
Issue number3
DOIs
StatePublished - Sep 1 2007

Fingerprint

Ehrlichia
Raccoons
Ticks
Cell Culture Techniques
Ixodes
Bacteria
Host Specificity
Endosomes
Ribosomal DNA
Epidemiologic Studies
Leukocytes
Erythrocytes
Cell Line
Polymerase Chain Reaction
Membranes
Antibodies

Keywords

  • Ehrlichia
  • Ixodes
  • Raccoon
  • Tick
  • Tick cell culture

Cite this

Isolation and establishment of the raccoon Ehrlichia-like agent in tick cell culture. / Munderloh, Ulrike G; Yabsley, Michael J.; Murphy, Staci M.; Luttrell, M. Page; Howerth, Elizabeth W.

In: Vector-Borne and Zoonotic Diseases, Vol. 7, No. 3, 01.09.2007, p. 418-425.

Research output: Contribution to journalArticle

Munderloh, Ulrike G ; Yabsley, Michael J. ; Murphy, Staci M. ; Luttrell, M. Page ; Howerth, Elizabeth W. / Isolation and establishment of the raccoon Ehrlichia-like agent in tick cell culture. In: Vector-Borne and Zoonotic Diseases. 2007 ; Vol. 7, No. 3. pp. 418-425.
@article{7ddb1249cc1042afad90da847d2fe48d,
title = "Isolation and establishment of the raccoon Ehrlichia-like agent in tick cell culture",
abstract = "Feral animals are reservoirs of emerging human pathogens, as well as carriers of closely related wildlife diseases. The latter may interfere with epidemiologic studies by inducing cross-reactive antibodies, or by providing false positive signals in PCR based tests. We cultured a novel intracellular bacterium from the blood of two raccoons (Procyon lotor): RAC413 and RAC414. RAC413 had been experimentally inoculated with blood from a wild-caught raccoon, and provided the material for a blood passage into RAC414. The microbes grew in Ixodes scapularis (black-legged tick) cells, line ISE6, inoculated either with the leukocyte or erythrocyte fraction of anticoagulated blood. Giemsa-stained cells sampled two and three months after initial inoculation of the cultures revealed inclusions similar to those of Ehrlichia sp., except that individual bacteria commonly were elongated and clustered within endosomes. Electronmicroscopy confirmed the presence of irregularly shaped bacteria with evenly granular bacterioplasm bounded by a unit membrane. 16S rDNA sequencing identified the microbes as the raccoon Ehrlichia-like agent previously detected in feral raccoons from Georgia, United States. In conclusion, the availability of a culture isolate of this agent will facilitate future studies to determine its biology, epidemiologic significance, vector association, and host range. The Ehrlichia-like agent infecting raccoons joins a growing list of tick-borne agents cultivable in tick cells.",
keywords = "Ehrlichia, Ixodes, Raccoon, Tick, Tick cell culture",
author = "Munderloh, {Ulrike G} and Yabsley, {Michael J.} and Murphy, {Staci M.} and Luttrell, {M. Page} and Howerth, {Elizabeth W.}",
year = "2007",
month = "9",
day = "1",
doi = "10.1089/vbz.2007.0640",
language = "English (US)",
volume = "7",
pages = "418--425",
journal = "Vector-Borne and Zoonotic Diseases",
issn = "1530-3667",
publisher = "Mary Ann Liebert Inc.",
number = "3",

}

TY - JOUR

T1 - Isolation and establishment of the raccoon Ehrlichia-like agent in tick cell culture

AU - Munderloh, Ulrike G

AU - Yabsley, Michael J.

AU - Murphy, Staci M.

AU - Luttrell, M. Page

AU - Howerth, Elizabeth W.

PY - 2007/9/1

Y1 - 2007/9/1

N2 - Feral animals are reservoirs of emerging human pathogens, as well as carriers of closely related wildlife diseases. The latter may interfere with epidemiologic studies by inducing cross-reactive antibodies, or by providing false positive signals in PCR based tests. We cultured a novel intracellular bacterium from the blood of two raccoons (Procyon lotor): RAC413 and RAC414. RAC413 had been experimentally inoculated with blood from a wild-caught raccoon, and provided the material for a blood passage into RAC414. The microbes grew in Ixodes scapularis (black-legged tick) cells, line ISE6, inoculated either with the leukocyte or erythrocyte fraction of anticoagulated blood. Giemsa-stained cells sampled two and three months after initial inoculation of the cultures revealed inclusions similar to those of Ehrlichia sp., except that individual bacteria commonly were elongated and clustered within endosomes. Electronmicroscopy confirmed the presence of irregularly shaped bacteria with evenly granular bacterioplasm bounded by a unit membrane. 16S rDNA sequencing identified the microbes as the raccoon Ehrlichia-like agent previously detected in feral raccoons from Georgia, United States. In conclusion, the availability of a culture isolate of this agent will facilitate future studies to determine its biology, epidemiologic significance, vector association, and host range. The Ehrlichia-like agent infecting raccoons joins a growing list of tick-borne agents cultivable in tick cells.

AB - Feral animals are reservoirs of emerging human pathogens, as well as carriers of closely related wildlife diseases. The latter may interfere with epidemiologic studies by inducing cross-reactive antibodies, or by providing false positive signals in PCR based tests. We cultured a novel intracellular bacterium from the blood of two raccoons (Procyon lotor): RAC413 and RAC414. RAC413 had been experimentally inoculated with blood from a wild-caught raccoon, and provided the material for a blood passage into RAC414. The microbes grew in Ixodes scapularis (black-legged tick) cells, line ISE6, inoculated either with the leukocyte or erythrocyte fraction of anticoagulated blood. Giemsa-stained cells sampled two and three months after initial inoculation of the cultures revealed inclusions similar to those of Ehrlichia sp., except that individual bacteria commonly were elongated and clustered within endosomes. Electronmicroscopy confirmed the presence of irregularly shaped bacteria with evenly granular bacterioplasm bounded by a unit membrane. 16S rDNA sequencing identified the microbes as the raccoon Ehrlichia-like agent previously detected in feral raccoons from Georgia, United States. In conclusion, the availability of a culture isolate of this agent will facilitate future studies to determine its biology, epidemiologic significance, vector association, and host range. The Ehrlichia-like agent infecting raccoons joins a growing list of tick-borne agents cultivable in tick cells.

KW - Ehrlichia

KW - Ixodes

KW - Raccoon

KW - Tick

KW - Tick cell culture

UR - http://www.scopus.com/inward/record.url?scp=35148824976&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=35148824976&partnerID=8YFLogxK

U2 - 10.1089/vbz.2007.0640

DO - 10.1089/vbz.2007.0640

M3 - Article

VL - 7

SP - 418

EP - 425

JO - Vector-Borne and Zoonotic Diseases

JF - Vector-Borne and Zoonotic Diseases

SN - 1530-3667

IS - 3

ER -