Detection of <i>Francisella tularensis</i>
 and analysis of bacterial growth in ticks in Japan

Suzuki, Junko; Hashino, Masanori; Matsumoto, Shogo; Takano, Ai; Kawabata, Hiroki; Takada, Nobuhiro; Andoh, Masako; Oikawa, Yosaburo; Kobayashi, Hiroko; Uda, Akihiko; Watanabe, Kenta; Shimizu, Takashi; Watarai, Masahisa

doi:10.1111/lam.12616

Cited by 18 publications

(11 citation statements)

References 37 publications

(57 reference statements)

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“…Ticks carry several pathogens including Rickettsia Japonica , 1 Rickettsia tamurae , 1 Rickettsia heilongiangensis , 2 Rickettsia helvetica , 1 Rickettsia asiatica , 1 Coxiella burneti 3 and Francisella tularensis 4 . There are no research reports on whether the invasion of pathogens carried by biting ticks on the human body has any effects to the human health or not.…”

Section: Figurementioning

confidence: 99%

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Investigation of pathogens injected into the human body by ticks

Kondo

Matsushima

Nakanishi

et al. 2022

Experimental Dermatology

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Ticks carry several pathogens including Rickettsia Japonica, 1Rickettsia tamurae, 1 Rickettsia heilongiangensis, 2 Rickettsia helvetica, 1 Rickettsia asiatica, 1 Coxiella burneti 3 and Francisella tularensis. 4 There are no research reports on whether the invasion of pathogens carried by biting ticks on the human body has any effects to the human health or not. In a non-endemic area of Japanese spotted fever in Mie Prefecture, Japan, the tick was removed with tweezers from three patients visited the hospital with complaint of being bitten by ticks. The removed ticks were soaked in 70% ethanol and stored.These patients had no fever and/or skin rash at the visit, and they remained in good general condition after tick removal without prophylactic antibiotic medication. The stocked ticks were extracted for DNA using a QIAamp DNA mini kit ® (Qiagen). Tick species were identified by PCR using widely used tick species identification prim-

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Section: Figurementioning

confidence: 99%

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confidence: 99%

Investigation of pathogens injected into the human body by ticks

Kondo

Matsushima

Nakanishi

et al. 2022

Experimental Dermatology

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“…Haemaphysalis flava ticks are widely distributed throughout southern China and many countries in Southeast Asia [1][2][3]. This tick species feeds on humans, cattle, boar, pandas and other mammals as common hematophagous ectoparasites and can carry bacteria, including Rickettsia, Anaplasma, Coxiella burnetti, Ehrlichia, Francisella tularensis, Borrelia theileri or Bartonella [4][5][6][7], and viruses, including severe fever with thrombocytopenia syndrome virus [8], tick-borne phleboviruses, Kabuto Mountain viruses [9] and Tarumizu tick viruses [10]. These pathogens are threats to human health and animal husbandry.…”

Section: Introductionmentioning

confidence: 99%

Comparative analysis of the anticoagulant activities and immunogenicity of HSC70 and HSC70TKD of Haemaphysalis flava

Liu

et al. 2022

Parasites Vectors

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Background Haemaphysalis flava is a hematophagous ectoparasite that acquires the nutrition needed for development and reproduction by sucking blood and digesting the blood meal. During blood-sucking and blood-meal digestion, the prevention of blood coagulation is important for this tick. Previous studies have shown that heat shock cognate 70 (HSC70) protein has certain anticoagulant activities, but its immunogenicity remains unclear. Also, whether the mutation of individual bases of the TKD-like peptide of HSC70 through the overlap extension method can change its anticoagulant activities and immunogenicity remains to be investigated. Methods The gene encoding the HSC70 protein was cloned from a complementary DNA library synthesized from H. flava. The coding gene of the TKD-like peptide of HSC70 was mutated into a TKD peptide coding gene (HSC70TKD) using the overlap extension method. Escherichia coli prokaryotic expression plasmids were constructed to obtain the recombinant proteins of HSC70 (rHSC70) and HSC70TKD (rHSC70TKD). The purified rHSC70 and rHSC70TKD were evaluated at different concentrations for anticoagulant activities using four in vitro clotting assays. Emulsifying recombinant proteins with complete and incomplete Freund’s adjuvants were subcutaneously immunized in Sprague Dawley rats. The serum antibody titers and serum concentrations of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were detected using an indirect enzyme-linked immunosorbent assay to assess the immunogenicity of rHSC70 and rHSC70TKD. Results The open reading frame of HSC70 was successfully amplified and found to have a length of 1958 bp. The gene encoding the TKD-like peptide of HSC70 was artificially mutated, with the 1373-position adenine (A) of the original sequence mutated into guanine (G), the 1385-position cytosine (C) mutated into G and the 1386-position G mutated into C. rHSC70 and rHSC70TKD that fused with His-tag were obtained using the expression plasmids pET-28a-HSC70 and pET-28a-HSC70TKD, respectively. rHSC70 and rHSC70TKD prolonged the thrombin time (TT) and reduced the fibrinogen (FIB) content in the plasma, but did not affect the prothrombin time (PT) or activated partial thromboplastin time (APTT) when compared to the negative control. Interestingly, the ability of rHSC70TKD to prolong the TT and reduce the FIB content in the plasma was better than that of rHSC70. The specific antibody titers of both rHSC70 and rHSC70TKD in rat serum reached 1:124,000 14 days after the third immunization. The serum concentration of IFN-γ in the rHSC70TKD group was higher than that in the rHSC70 group. The rHSC70 group has the highest serum concentration of IL-4, and the serum concentration of IL-4 in the rHSC70TKD group was higher than that in the negative group. Conclusions rHSC70 and rHSC70TKD exhibited anticoagulant activities by prolonging the TT and reducing the FIB content in vitro. rHSC70TKD had better anticoagulant activities than rHSC70. Both rHSC70 and rHSC70TKD had good immunogenicity and induced humoral and cellular immunity.

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“…A variety of pathogens are detected in those ticks, such as encephalitis virus (Ko et al ., ), Rickettsiae spp. (Lu et al ., ), Kabuto Mountain virus (Ejiri et al ., ), Tarumizu tick virus (Fujita et al ., ), Francisella tularensis (Suzuki et al ., ) and Cercopithifilaria spp. (Uni et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Cloning of four HSPA multigene family members in Haemaphysalis flava ticks

Liu

et al. 2019

Medical Vet Entomology

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The heat shock protein 70 (HSPA) family and their genes have been studied in ticks and are considered as possible antigen candidates for the development of anti-tick vaccines. However, knowledge about their members, structure and function in ticks is incomplete. Based on our transcriptomic data, the full length of four HSPA genes in Haemaphysalis flava (Acari: Ixodidae) was cloned via rapid amplification of cDNA ends. The open reading frame of HSPA2A, HSPA2B, HSPA5 and HSPA9 was 1920, 1911, 1983 and 2088 bp in length, respectively. Three family signatures and one localization motif were in the encoding proteins. HSPA2A and HSPA2B were predicted to be located at cytoplasm/nucleus, whereas HSPA5 and HSPA9 were at endoplasmic reticulum and mitochondria, respectively. In silico simulation demonstrated that those proteins had distinct numbers of -helixes, extended strands and coils, and different antigenic epitopes. Expression of HSPA5 and HSPA9 in the salivary gland was significantly higher in partially-engorged female adult ticks than the fully-engorged (P < 0.01) as shown by a quantitative polymerase chain reaction. Our data indicated that H. flava ticks had at least four HSPA genes encoding proteins with different cellular locations, structures and expression profiles, suggesting their diverse roles in tick biology.

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Detection of Francisella tularensis and analysis of bacterial growth in ticks in Japan

Cited by 18 publications

References 37 publications

Investigation of pathogens injected into the human body by ticks

Investigation of pathogens injected into the human body by ticks

Comparative analysis of the anticoagulant activities and immunogenicity of HSC70 and HSC70TKD of Haemaphysalis flava

Cloning of four HSPA multigene family members in Haemaphysalis flava ticks

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