scholarly journals Identification of a Granulocytic Ehrlichia Strain Isolated from a Horse in Switzerland and Comparison with Other Rickettsiae of the Ehrlichia phagocytophilaGenogroup

1998 ◽  
Vol 36 (7) ◽  
pp. 2035-2037 ◽  
Author(s):  
Nicola Pusterla ◽  
Jon B. Huder ◽  
Karsten Feige ◽  
Hans Lutz
Keyword(s):  
Case Report ◽  
Nucleotide Sequence ◽  
16S Rrna ◽  
Clinical Signs ◽  
Buffy Coat ◽  
Rrna Gene ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Human Granulocytic Ehrlichiosis ◽  
Limb Edema

This case report describes a 12-year-old Arabian mare with granulocytic ehrlichiosis. Clinical signs included fever, apathy, anorexia, icterus, limb edema, and reluctance to move. Examination of buffy coat smears revealed Ehrlichia organisms in neutrophils and eosinophils. A band of 1,428 bp was amplified from DNA of leukocytes via nested PCR and was identified as part of theEhrlichia 16S rRNA gene. It differed from the gene sequences of Ehrlichia phagocytophila and E. equi at two and three positions, respectively. Interestingly, the nucleotide sequence of the 16S rRNA was 100% identical to that of the agent of human granulocytic ehrlichiosis.

Improved Sensitivity of PCR for Diagnosis of Human Granulocytic Ehrlichiosis Using epank1 Genes of Ehrlichia phagocytophila -Group Ehrlichiae

2000 ◽  
Vol 38 (1) ◽  
pp. 354-356
Author(s):  
Jennifer J. Walls ◽  
Patrizio Caturegli ◽  
Johan S. Bakken ◽  
Kristin M. Asanovich ◽  
J. Stephen Dumler
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Ankyrin Repeat ◽  
Rrna Gene ◽  
Gene Target ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Human Granulocytic Ehrlichiosis ◽  
Group A ◽  
Group B

ABSTRACT The agent of human granulocytic ehrlichiosis (HGE), Ehrlichia phagocytophila , and Ehrlichia equi probably comprise variants of a single Ehrlichia species now called the Ehrlichia phagocytophila genogroup. These variants share a unique 153-kDa protein antigen with ankyrin repeat motifs encoded by the epank1 gene. The epank1 gene was investigated as an improved target for PCR diagnosis of HGE compared with the currently used 16S rRNA gene target. Primers for epank1 flanking a region that spans part of the 5′ ankyrin repeat coding region and part of the unique 3′ region were synthesized. Blood samples from 31 patients with suspected HGE who were previously tested by 16S rRNA gene (16S) PCR and indirect immunofluorescent antibody test (IFA) were retrospectively tested with the epank1 primers. Eleven patients were 16S PCR positive and had a seroconversion detected by IFA (group A), 10 patients were 16S PCR negative but had a seroconversion detected by IFA (group B), and 10 patients were 16S PCR negative and seronegative (group C). Ten of the 11 group A patients were epank1 PCR positive, all 10 of the group B patients were epank1 PCR positive, and all of the PCR-negative and seronegative patients (group C) were epank1 PCR negative. The epank1 primers are more sensitive than the previously used 16S rRNA gene primers and therefore may be more useful in diagnostic testing for HGE.

scholarly journals Human Granulocytic Ehrlichiosis Agent Infection in a Pony Vaccinated with a Borrelia burgdorferi Recombinant OspA Vaccine and Challenged by Exposure to Naturally Infected Ticks

2000 ◽  
Vol 7 (1) ◽  
pp. 68-71 ◽  
Author(s):  
Yung-Fu Chang ◽  
Sean P. McDonough ◽  
Chao-Fu Chang ◽  
Kwang-Soon Shin ◽  
William Yen ◽  
...  
Keyword(s):  
Borrelia Burgdorferi ◽  
Inclusion Bodies ◽  
Ixodes Scapularis ◽  
Clinical Signs ◽  
Buffy Coat ◽  
The Body ◽  
Granulocytic Ehrlichiosis ◽  
Flunixin Meglumine ◽  
Human Granulocytic Ehrlichiosis ◽  
Vector Dna

ABSTRACT A pony was vaccinated with recombinant OspA vaccine (rOspA) and then exposed 3 months later to Borrelia burgdorferi-infected ticks (Ixodes scapularis) collected in Westchester County, N.Y. At 2 weeks after tick exposure, the pony developed a high fever (105°F). Buffy coat smears showed that 20% of neutrophils contained ehrlichial inclusion bodies (morulae). Flunixin Meglumine (1 g daily) was given for 2 days, and the body temperature returned to normal. PCR for ehrlichial DNA was performed on blood samples for 10 consecutive days beginning when the pony was first febrile. This pony was monitored for another 3.5 months but developed no further clinical signs. The 44-kDa immunodominant human granulocytic ehrlichiosis antigen gene was amplified by PCR and cloned into a pCR2.1 vector. DNA sequence analysis of this gene showed it was only 8 bp different (99% identity) from the results reported by others (J.W. Ijdo et al., Infect. Immun. 66:3264–3269, 1998). Western blot analysis, growth inhibition assays, and repeated attempts to isolate B. burgdorferi all demonstrated the pony was protected against B. burgdorferi infection. These results highlight the potential for ticks to harbor and transmit several pathogens simultaneously, which further complicates the diagnosis and vaccination of these emerging tick-borne diseases.

scholarly journals Evidence of the Human Granulocytic Ehrlichiosis Agent in Ixodes ricinus Ticks in Switzerland

1999 ◽  
Vol 37 (5) ◽  
pp. 1332-1334 ◽  
Author(s):  
Nicola Pusterla ◽  
Christian M. Leutenegger ◽  
Jon B. Huder ◽  
Rainer Weber ◽  
Ueli Braun ◽  
...  
Keyword(s):  
Ixodes Ricinus ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Adult Males ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Stage Of Development ◽  
Human Granulocytic Ehrlichiosis ◽  
Pcr Products ◽  
Ixodes Ricinus Ticks

A total of 1,667 Ixodes ricinus ticks were collected from five regions in Switzerland where there have been sporadic occurrences of granulocytic ehrlichiosis in dogs and horses. The ticks were examined for rickettsiae of the Ehrlichia phagocytophila group via nested PCR. Twenty-one ticks (1.3%) were positive; 3 (0.5%) were nymphs, 6 (1.3%) were adult males, and 12 (1.9%) were adult females. The number of positive ticks varied with the stage of development and with the geographical origin. Nucleotide sequencing of the isolated PCR products identified these products as part of the 16S rRNA gene of Ehrlichia. In addition, these products had 100% homology with the agent of human granulocytic ehrlichiosis. The occurrence of this agent in I. ricinus in Switzerland presents a potential danger of transmission of granulocytic ehrlichiosis to dogs, horses, and humans.

scholarly journals Isolation and Characterization of Two European Strains of Ehrlichia phagocytophila of Equine Origin

2002 ◽  
Vol 9 (2) ◽  
pp. 341-343 ◽  
Author(s):  
Anneli Bjöersdorff ◽  
Bodil Bagert ◽  
Robert F. Massung ◽  
Asiya Gusa ◽  
Ingvar Eliasson
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Isolation And Characterization ◽  
Operon Gene ◽  
The 16S Rrna Gene

ABSTRACT We report the isolation and partial genetic characterization of two equine strains of granulocytic Ehrlichia of the genogroup Ehrlichia phagocytophila. Frozen whole-blood samples from two Swedish horses with laboratory-verified granulocytic ehrlichiosis were inoculated into HL-60 cell cultures. Granulocytic Ehrlichia was isolated and propagated from both horses. DNA extracts from the respective strains were amplified by PCR using primers directed towards the 16S rRNA gene, the groESL heat shock operon gene, and the ank gene. The amplified gene fragments were sequenced and compared to known sequences in the GenBank database. With respect to the 16S rRNA gene, the groESL gene, and the ank gene, the DNA sequences of the two equine Ehrlichia isolates were identical to sequences found in isolates from clinical cases of granulocytic ehrlichiosis in humans and domestic animals in Sweden. However, compared to amplified DNA from an American Ehrlichia strain of the E. phagocytophila genogroup, differences were found in the groESL gene and ank gene sequences.

scholarly journals Experimental Transmission of Ehrlichia equi to Horses through Naturally Infected Ticks (Ixodes pacificus) from Northern California

1998 ◽  
Vol 36 (7) ◽  
pp. 2131-2134 ◽  
Author(s):  
Gerhard H. Reubel ◽  
Robert B. Kimsey ◽  
Jeffrey E. Barlough ◽  
John E. Madigan
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Clinical Signs ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Ixodes Pacificus ◽  
Experimental Transmission ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Equi

We report the experimental transmission of Ehrlichia equi from naturally infected Ixodes pacificus ticks to horses. Three weeks after exposure to ticks, two of three horses developed clinical signs compatible with E. equiinfection, while one horse remained asymptomatic. 16S rRNA gene PCR of blood leukocyte lysates was positive for all horses at various time points; two horses seroconverted. The 16S rRNA gene sequences amplified from tick-exposed horses showed more than 99% homology to corresponding fragments of the 16S rRNA genes of E. equi, Ehrlichia phagocytophila, and the human granulocytic ehrlichiosis agent.

scholarly journals Comparison of the Nucleotide Sequences of 16S rRNA, 444Ep-ank, and groESL Heat Shock Operon Genes in Naturally Occurring Ehrlichia equi and Human Granulocytic Ehrlichiosis Agent Isolates from Northern California

2000 ◽  
Vol 38 (4) ◽  
pp. 1364-1369 ◽  
Author(s):  
Joon-seok Chae ◽  
Janet E. Foley ◽  
J. Stephen Dumler ◽  
John E. Madigan
Keyword(s):  
Heat Shock ◽  
16S Rrna ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Northern California ◽  
Eastern United States ◽  
Granulocytic Ehrlichiosis ◽  
Naturally Occurring ◽  
Human Granulocytic Ehrlichiosis ◽  
Ehrlichia Equi

We examined 11 naturally occurring isolates of Ehrlichia equi in horses and two human granulocytic ehrlichiosis agent isolates in California for sequence diversity in three genes.Ehrlichia equi isolates were from Sierra (n = 6), Mendocino (n = 3), Sonoma (n = 1), and Marin (n = 1) counties, and human granulocytic ehrlichiosis (HGE) agent isolates were obtained from Humboldt county. PCR with specific primers for 16S rRNA, 444Ep-ank and groESL heat shock operon genes successfully produced amplicons for all 13 clinical samples. The 444Ep-ank gene of the HGE agent and E. equiisolates from northern California is different from the eastern U.S. isolates BDS and USG3. The translated amino acid sequence of thegroESL heat shock operon gene fragment is identical amongE. equi, the HGE agent, and E. phagocytophila, with the exception of the northern Californian equine CASOLJ isolate. Microheterogeneity was observed in the 16S rRNA gene sequences of HGE agent and E. equi isolates from northern California. These results suggest that E. equi and the HGE agent found in California are similar or identical but may differ from the isolates of equine and human origin found in the eastern United States.

scholarly journals Identity of Ehrlichial DNA Sequences Derived from Ixodes ricinus Ticks with Those Obtained from Patients with Human Granulocytic Ehrlichiosis in Slovenia

1999 ◽  
Vol 37 (1) ◽  
pp. 209-210 ◽  
Author(s):  
M. Petrovec ◽  
J. W. Sumner ◽  
W. L. Nicholson ◽  
J. E. Childs ◽  
F. Strle ◽  
...  
Keyword(s):  
Heat Shock ◽  
16S Rrna ◽  
Ixodes Ricinus ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Granulocytic Ehrlichiosis ◽  
Human Granulocytic Ehrlichiosis ◽  
Ixodes Ricinus Ticks ◽  
Pcr Assays ◽  
The 16S Rrna Gene

Adult Ixodes ricinus (Acari: Ixodidae) ticks collected near Ljubljana, Slovenia, were tested for the agent of human granulocytic ehrlichiosis (HGE) by using PCR assays based on the 16S rRNA gene. Three (3.2%) of 93 ticks were found to contain granulocytic ehrlichiae. Nucleotide sequences of portions of the bacterialgroESL heat shock operon amplified from these ticks were identical or nearly (99.8%) identical to those previously determined for human patients with HGE from Slovenia, providing additional evidence that the ticks were infected with the HGE agent. This study identified I. ricinus as the likely vector for these ehrlichial pathogens of humans in this part of Europe.

scholarly journals Quantitative Real-Time PCR for Detection of Members of the Ehrlichia phagocytophila Genogroup in Host Animals and Ixodes ricinus Ticks

1999 ◽  
Vol 37 (5) ◽  
pp. 1329-1331 ◽  
Author(s):  
Nicola Pusterla ◽  
Jon B. Huder ◽  
Christian M. Leutenegger ◽  
Ueli Braun ◽  
John E. Madigan ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Ixodes Ricinus ◽  
Nested Pcr ◽  
Rrna Gene ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Ixodes Ricinus Ticks ◽  
Taqman Pcr ◽  
The 16S Rrna Gene

A TaqMan PCR was established for identification and quantitation of members of the Ehrlichia phagocytophila group in experimentally infected cows and in Ixodes ricinus ticks. The TaqMan PCR identified a 106-bp section of the 16S rRNA gene by use of a specific fluorogenic probe and two primers. This technique was specific for members of the E. phagocytophila group, which include E. phagocytophila, Ehrlichia equi, and the agent of human granulocytic ehrlichiosis. The TaqMan system identified 10 copies of a cloned section of the 16S rRNA gene ofE. phagocytophila. The sensitivity and specificity of the TaqMan PCR were similar to those of conventional nested PCR. The numbers of ehrlichiae in leukocytes of the two cows experimentally infected with E. phagocytophila were measured daily by TaqMan PCR and had a course similar to that of the percentages of infected leukocytes determined daily by light microscopy. The prevalence of infected free-living ticks, which were collected from areas where bovine ehrlichiosis is endemic and from regions with sporadic occurrences of granulocytic ehrlichiosis in dogs and horses, was identical as determined by nested PCR and TaqMan PCR.

scholarly journals Antimicrobial Susceptibility ofEhrlichia phagocytophila

2001 ◽  
Vol 45 (3) ◽  
pp. 786-788 ◽  
Author(s):  
Harold W. Horowitz ◽  
T.-C. Hsieh ◽  
Maria E. Aguero-Rosenfeld ◽  
Fatemeh Kalantarpour ◽  
Ishraq Chowdhury ◽  
...  
Keyword(s):  
New York ◽  
Culture System ◽  
Minimum Bactericidal Concentration ◽  
New York State ◽  
Cell Culture System ◽  
Obligate Intracellular ◽  
Granulocytic Ehrlichiosis ◽  
Ehrlichia Phagocytophila ◽  
Human Granulocytic Ehrlichiosis ◽  
Quinolone Antibiotics

ABSTRACT Human granulocytic ehrlichiosis is a recently described disease caused by an obligate intracellular gram-negative organism recently named Ehrlichia phagocytophila. To expand our knowledge of the susceptibility of E. phagocytophila, we tested six New York State isolates for susceptibility to 12 antimicrobials using an HL-60 cell culture system. All of the isolates were susceptible to doxycycline (MIC, ≤0.125 μg/ml; minimum bactericidal concentration [MBC], 0.125 to 0.5 μg/ml), rifampin (MIC, ≤0.125 μg/ml; MBC, ≤0.125 μg/ml), ofloxacin (MIC, ≤2 μg/ml; MBC, ≤2 μg/ml), levofloxacin (MIC, ≤1 μg/ml; MBC, ≤1 μg/ml), and trovafloxacin (MIC, ≤0.032 μg/ml; MBC, ≤0.032 μg/ml). Isolates were uniformly resistant to amoxicillin, ceftriaxone, erythromycin, azithromycin, clarithromycin, and amikacin. For one strain, the MBC of chloramphenicol was ≤8 μg/ml. These data suggest that quinolone antibiotics and rifampin may be alternative agents for patients with intolerance to tetracyclines.

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