Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

Murase, Yusuke; Konnai, Satoru; Hidano, Arata; Githaka, Naftaly; Ito, Takuya; Takano, Ai; Kawabata, Hiroki; Ato, Manabu; Tajima, Tomoko; Tajima, Motoshi; Onuma, Misao; Shiro, Motoo; Ohashi, Kazuhiko

doi:10.1016/j.vetmic.2010.11.025

Cited by 24 publications

(15 citation statements)

References 15 publications

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“…in the present study (27.5% in all ticks, or 26.9% if only I. persulcatus is considered) is higher than the previously reported rate of 7.1% in I. persulcatus ticks [24]. Prevalence rates of A. phagocytophilum in cattle of 1.0-3.4% have also been reported [15,24]. However, the previous reports used different PcR methods, which may also differ in sensitivities or specificities, leading to varying results.…”

Section: Discussioncontrasting

confidence: 84%

“…closely related to A. phagocytophilum in Japan. In a related study, samples found to be Anaplasma-positive using a 16S rRNA-based PcR revealed negative results using a PcR based on p44/ msp2 [24]. Thus, the new reliable PcR method developed in this study can be used to clarify results as well as to conduct further studies to assess the epidemiological status of the 2 organisms in Japan.…”

Section: Discussionmentioning

confidence: 68%

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Dual Presence of Anaplasma phagocytophilum and Its Closely Related Anaplasma sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

Ybañez

Matsumoto

Kishimoto

et al. 2012

The Journal of Veterinary Medical Science

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ABSTRAcT. Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) and tick-borne fever in ruminants. A closely related and potentially novel Anaplasma sp. in Japan was recently characterized. The aims of the study were to provide molecular evidence for the presence of these 2 species in Japan, and to develop a reliable PcR method based on the nucleotide differences within the citrate synthase (gltA) gene. DNA samples from 182 ixodid ticks (134 Ixodes persulcatus, 35 Haemaphysalis douglasii and 13 I. ovatus) collected from 2 sites in Hokkaido, Japan, were screened for A. phagocytophilum and its closely related Anaplasma sp. (herein designated as Anaplasma sp. Japan) using 16S rRNA PcR, revealing a combined prevalence rate of 27.5% (50 samples). The positive samples were then used to evaluate a newly developed gltA-based nested PcR method. Selected positive samples were further characterized using the groEL gene for confirmation and phylogenetic analyses. Two groups of sequence results were obtained: those that had closer identities with (1) A. phagocytophilum (99.5-99.6% for 16S rRNA, 97.5% for gltA and 98.4% for groEL), and those that had closer identities with (2) Anaplasma sp. closely related to A. phagocytophilum in Japan (99.3% for 16S rRNA, 96.4-98.7% for gltA and 97.5-97.9% for groEL). The present study confirmed the distinct presence of A. phagocytophilum and its closely related Anaplasma sp. in Japan, and developed a new PcR detection method based on gltA that can distinguish the 2 organisms.

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Section: Discussioncontrasting

confidence: 84%

Section: Discussionmentioning

confidence: 68%

Dual Presence of Anaplasma phagocytophilum and Its Closely Related Anaplasma sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

Ybañez

Matsumoto

Kishimoto

et al. 2012

The Journal of Veterinary Medical Science

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“…Lyme borrelia and A. phagocytophilum, the agent of human granulocytic anaplasmaposis, are both transmitted by Ixodes spp., including I. persulcatus and may occasionally co-infect a host. In our previous study [18], out of the 281 I. persulcatus ticks, twenty (7.1%) were found to harbor A. phagocytophilum DNA. Finally, the co-infection rate for B. garinii and A. phagocytophilum in I. persulcatus ticks was 3.9% (11/284).…”

confidence: 83%

“…Of the I. persulcatus ticks from the field, 12 ticks (4.2%) had mixed infection with B. garinii and B. afzelii. Recently, we reported an epidemiological surveillance of Anaplasma phagocytophilum in I. persulcatus ticks in Hokkaido [18]. As in the present study, DNA samples extracted from I. persulcatus had been used to detect A. phagocytophilum by PCR with primers MSP2-3F (5'-CCA GCG TTT AGC AAG ATA AGA G-3') and MSP2-3R (5'-GCC CAG TAA CAA CAT CAT AAG C-3') as described previously [13,18,27], which can specifically amplify a 334 bp portion of the p44 gene of A. phagocytophilum.…”

confidence: 99%

Prevalence of Lyme Borrelia in Ixodes persulcatus Ticks from an Area with a Confirmed Case of Lyme Disease

et al. 2013

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ABSTRACT. In this study, the prevalence of Borrelia infections in Ixodes ticks from a site in Hokkaido, Japan, with confirmed cases of Lyme disease was determined by a PCR method capable of detecting and differentiating between strains of pathogenic Borrelia, with particular emphasis on Borrelia garinii (B. garinii) and Borrelia afzelli (B. afzelli), using tick-derived DNA extracts as template. A total of 338 ticks, inclusive of 284 Ixodes persulcatus (I. persulcatus), were collected by flagging vegetation in mid-spring. Ninety-eight (34.5%) of I. persulcatus tested positive for Borrelia species DNA, whereas the overall prevalence of Borrelia species in Ixodes ovatus and Haemaphysalis longicornis ticks was 19.5 and 7.7%, respectively. PCR-RFLP and sequence analysis of Borrelia rrf(5S)-rrl(23S) intergenic spacer DNA amplicons indicated that they originated from three different Borrelia species namely, B. garinii, B. afzelii and B. japonica. Among the I. persulcatus species, which is a known vector of human borreliosis, 86 were mono-infected with B. garinii, 2 ticks were mono-infected with B. afzelii and whereas 12 ticks had dual infections. Most significant, 11 of the I. persulcatus ticks were coinfected with Anaplasma phagocytophilum and B. garinii. The difference between the number of obtained and expected co-infections was significant (χ 2 =4.32, P=0.038).

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“…Although Ixodes ticks often feed on white-tailed deer, the deer are infected with the Ap-Variant 1 strain of A. phagocytophilum, rather than with the human strain, in the United States (146). Diverse A. phagocytophilum strains are also found in animals and ticks in Europe, Japan, and Russia (109,111,151,160,170,203,238,239), where HGA has been rarely reported. These findings imply that the zoonosis potential of A. phagocytophilum depends not only on the transmissibility, habitats, and population density of ticks and infected mammals (90) but also on the genetic variations of A. phagocytophilum.…”

Section: Morphologymentioning

confidence: 99%

Mechanisms of Obligatory Intracellular Infection with Anaplasma phagocytophilum

2011

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SUMMARY Anaplasma phagocytophilum persists in nature by cycling between mammals and ticks. Human infection by the bite of an infected tick leads to a potentially fatal emerging disease called human granulocytic anaplasmosis. A. phagocytophilum is an obligatory intracellular bacterium that replicates inside mammalian granulocytes and the salivary gland and midgut cells of ticks. A. phagocytophilum evolved the remarkable ability to hijack the regulatory system of host cells. A. phagocytophilum alters vesicular traffic to create an intracellular membrane-bound compartment that allows replication in seclusion from lysosomes. The bacterium downregulates or actively inhibits a number of innate immune responses of mammalian host cells, and it upregulates cellular cholesterol uptake to acquire cholesterol for survival. It also upregulates several genes critical for the infection of ticks, and it prolongs tick survival at freezing temperatures. Several host factors that exacerbate infection have been identified, including interleukin-8 (IL-8) and cholesterol. Host factors that overcome infection include IL-12 and gamma interferon (IFN-γ). Two bacterial type IV secretion effectors and several bacterial proteins that associate with inclusion membranes have been identified. An understanding of the molecular mechanisms underlying A. phagocytophilum infection will foster the development of creative ideas to prevent or treat this emerging tick-borne disease.

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Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

Cited by 24 publications

References 15 publications

Dual Presence of <i>Anaplasma phagocytophilum</i> and Its Closely Related <i>Anaplasma</i> sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

Dual Presence of <i>Anaplasma phagocytophilum</i> and Its Closely Related <i>Anaplasma</i> sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

Prevalence of Lyme Borrelia in <i>Ixodes persulcatus</i> Ticks from an Area with a Confirmed Case of Lyme Disease

Mechanisms of Obligatory Intracellular Infection with Anaplasma phagocytophilum

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