Detection of four Borrelia burgdorferi genospecies and first report of human granulocytic ehrlichiosis agent in Ixodes ricinus ticks collected in central Italy

The presence of Borrelia burgdorferi s.l. and of Ehrlichia phagocytophila group was sought by PCR in Ixodes ricinus collected in a protected area of central Italy. Nymphs (n = 1475, gathered in 295 pools of 5 nymphs each) and adult ticks (n = 28) were examined. B. burgdorferi s.l. was detected in 13.8% of the nymph pools; of these, 63.4% were infected by B. valaisiana, 26.8% by B. afzelii, 7.3% by B. garinii, and 2.5% by B. burgdorferi s.s. Only a single adult male tick proved to host B. afzelii. The agent of human granulocytic ehrlichiosis (HGE) was detected in 2.7% of the nymph pools. Two HGE agent-positive nymph pools were also found to be positive for B. garinii and for B. afzelii, respectively. This is the first report from central Italy of the finding of the HGE agent in ticks.

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

Journal of Clinical Microbiology ◽

10.1128/jcm.37.5.1332-1334.1999 ◽

1999 ◽

Vol 37 (5) ◽

pp. 1332-1334 ◽

Cited By ~ 24

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.

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Prevalence of Borrelia Burgdorferi Sensu Lato Genomospecies and of the Human Granulocytic Ehrlichiosis (HGE) Agent in Ixodes Ricinus Ticks Collected in the Area of Monti Lepini, Italy

International Journal of Immunopathology and Pharmacology ◽

10.1177/039463200301600203 ◽

2003 ◽

Vol 16 (2) ◽

pp. 105-108 ◽

Cited By ~ 10

Author(s):

I. Santino ◽

A. Iori ◽

M. Nicoletti ◽

S. Valletta ◽

C. Cimmino ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Pcr Amplification ◽

Sensu Stricto ◽

16S Rrna Genes ◽

Rrna Genes ◽

Borrelia Burgdorferi Sensu Lato ◽

Granulocytic Ehrlichiosis ◽

Ehrlichia Phagocytophila ◽

Human Granulocytic Ehrlichiosis ◽

Ixodes Ricinus Ticks

Ticks are obligate hematophagous arthropods that are parasites in every class of vertebrates in most regions of the world. They are also considered to be important vectors for the transmission of human infectious diseases. In the present study we used polymer chain reaction (PCR) amplification analysis to determine the prevalence of Borrelia burgdorferi and Ehrlichia phagocytophila, the agents of, respectively, Lyme borreliosis and human granulocytic ehrlichiosis, among ticks inhabiting the area of Monti Lepini, a wild area located in the Latium Region of Italy. A total of 141 I. ricinus ticks (125 nymphs and 16 adults) were collected in the studied area. Total DNAs were extracted from I. ricinus nymphs (pooled in groups of five) and from individual adults. The DNA samples were examined for the presence of B. burgdorferi sensu lato and E. phagocytophila by PCR using two specific pairs of oligonucleotides that specifically amplify distinct DNA regions of the 16S rRNA genes of the two species. The prevalence of vectors infected with B. burgdorferi s. 1. was 16% in pooled nymphs samples, and 12.5% in adult ticks, while E. phagocytophila was found only in pooled nymphs samples (8%). Three genomospecies were identified, namely Borrelia afzelii, Borrelia garinii, and Borrelia valaisiana, in samples found positive for B. burgdorferi s. 1. No sample was found positive for Borrelia burgdorferi sensu stricto.

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Acquisition of Coinfection and Simultaneous Transmission of Borrelia burgdorferi and Ehrlichia phagocytophila by Ixodes scapularis Ticks

Infection and Immunity ◽

10.1128/iai.68.4.2183-2186.2000 ◽

2000 ◽

Vol 68 (4) ◽

pp. 2183-2186 ◽

Cited By ~ 72

Author(s):

Michael L. Levin ◽

Durland Fish

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Ixodes Scapularis ◽

The Other ◽

Infection Status ◽

Granulocytic Ehrlichiosis ◽

Ehrlichia Phagocytophila ◽

Human Granulocytic Ehrlichiosis ◽

Previous Infection ◽

Prior Infection

ABSTRACT The agents of Lyme disease (Borrelia burgdorferi) and human granulocytic ehrlichiosis (Ehrlichia phagocytophila) are both transmitted by the tick Ixodes scapularis. In nature, ticks are often infected with both agents simultaneously. We studied whether previous infection with either Borrelia orEhrlichia in ticks would affect acquisition and transmission of a second pathogen. Ehrlichia-infectedI. scapularis nymphs were fed uponBorrelia-infected mice, and Borrelia-infectedI. scapularis nymphs were fed uponEhrlichia-infected mice. The efficiency with which previously infected nymphal ticks acquired a second pathogen from infected hosts was compared to that of uninfected ticks. An average of 51% ± 15% of ticks acquired Ehrlichia from infected mice regardless of their prior infection status with Borrelia. An average of 85% ± 10% of ticks acquired Borrelia from infected mice regardless of their prior infection status withEhrlichia. Also, we assessed the efficiency with which individual nymphs could transmit either agent alone, or both agents simultaneously, to individual susceptible hosts. An average of 76% ± 9% of Borrelia-infected ticks and 84% ± 10% ofEhrlichia-infected ticks transmitted these agents to mice regardless of the presence of the other pathogen. There was no evidence of interaction between the agents of Lyme disease and human granulocytic ehrlichiosis in I. scapularis ticks. The presence of either agent in the ticks did not affect acquisition of the other agent from an infected host. Transmission of the agents of Lyme disease and human granulocytic ehrlichiosis by individual ticks was equally efficient and independent. Dually infected ticks transmitted each pathogen to susceptible hosts as efficiently as ticks infected with only one pathogen.

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Prevalence of Borrelia burgdorferi and Granulocytic and Monocytic Ehrlichiae in Ixodes ricinusTicks from Southern Germany

Journal of Clinical Microbiology ◽

10.1128/jcm.37.11.3448-3451.1999 ◽

1999 ◽

Vol 37 (11) ◽

pp. 3448-3451 ◽

Cited By ~ 52

Author(s):

Birgit U. Baumgarten ◽

Martin Röllinghoff ◽

Christian Bogdan

Keyword(s):

Borrelia Burgdorferi ◽

The United States ◽

Tick Population ◽

Original Sequence ◽

Southern Germany ◽

Granulocytic Ehrlichiosis ◽

Ehrlichia Phagocytophila ◽

16S Ribosomal Dna ◽

Human Granulocytic Ehrlichiosis ◽

Pcr Products

A total of 287 adult Ixodes ricinus ticks, collected in two regions of southern Germany (Frankonia and Baden-Württemberg) where Borrelia burgdorferi infections are known to be endemic, were examined for the presence of 16S ribosomal DNA specific for the Ehrlichia phagocytophila genogroup, E. chaffeensis, E. canis, and B. burgdorferiby nested PCR. Totals of 2.2% (6 of 275) and 21.8% (65 of 275) of the ticks were positive for the E. phagocytophila genogroup andB. burgdorferi, respectively. Two ticks (0.7%) were coinfected with both bacteria. Of 12 engorged I. ricinusticks collected from two deer, 8 (67%) were positive for the E. phagocytophila genogroup and one (8%) was positive for B. burgdorferi. There was no evidence of infection with E. canis or E. chaffeensis in the investigated tick population. The nucleotide sequences of the 546-bpEhrlichia PCR products differed at one or two positions from the original sequence of the human granulocytic ehrlichiosis (HGE) agent (S.-M. Chen, J. S. Dumler, J. S. Bakken, and D. H. Walker, J. Clin. Microbiol. 32:589–595, 1994). Three groups of sequence variants were detected; two of these were known to occur in other areas in Europe or the United States, whereas one has not been reported before. Thus, in the German I. ricinus tick population closely related granulocytic ehrlichiae are prevalent, which might represent variants of E. phagocytophila or the HGE agent.

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

Journal of Clinical Microbiology ◽

10.1128/jcm.37.1.209-210.1999 ◽

1999 ◽

Vol 37 (1) ◽

pp. 209-210 ◽

Cited By ~ 45

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.

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

Journal of Clinical Microbiology ◽

10.1128/jcm.37.5.1329-1331.1999 ◽

1999 ◽

Vol 37 (5) ◽

pp. 1329-1331 ◽

Cited By ~ 51

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.

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Molecular Evidence of Coinfection of Ticks with Borrelia burgdorferi Sensu Lato and the Human Granulocytic Ehrlichiosis Agent in Switzerland

Journal of Clinical Microbiology ◽

10.1128/jcm.37.10.3390-3391.1999 ◽

1999 ◽

Vol 37 (10) ◽

pp. 3390-3391 ◽

Cited By ~ 47

Author(s):

Christian M. Leutenegger ◽

Nicola Pusterla ◽

Caroline N. Mislin ◽

Rainer Weber ◽

Hans Lutz

Keyword(s):

Borrelia Burgdorferi ◽

Real Time ◽

Ixodes Ricinus ◽

Real Time Pcr ◽

Borrelia Burgdorferi Sensu Lato ◽

Molecular Evidence ◽

Granulocytic Ehrlichiosis ◽

Human Granulocytic Ehrlichiosis

Adult Ixodes ricinus ticks were collected in Switzerland and tested for the presence of coinfection withBorrelia burgdorferi sensu lato and the human granulocytic ehrlichiosis (HGE) agent by real-time PCR. Of 100 ticks, 49% were positive for B. burgdorferi and 2% were positive for the HGE agent. The two HGE agent-positive ticks were also found to be positive for B. burgdorferi.

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