Babesia gibsoni: Detection in blood smears and formalin-fixed, paraffin-embedded tissues using deoxyribonucleic acid in situ hybridization analysis

Yamasaki, Masahiro; Kobayashi, Yusuke; Nakamura, Kensuke; Sasaki, Noboru; Murakami, Masahiro; Rajapakshage, Bandula Kumara Wickramasekara; Ohta, Hiroshi; Yamato, Osamu; Maede, Yoshimitsu; Takiguchi, Mitsuyoshi

doi:10.1016/j.exppara.2010.07.004

Cited by 6 publications

(4 citation statements)

References 25 publications

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“…(Yamasaki et al . ). For ISH analysis, blood smears were formalin‐fixed and paraffin‐embedded before mounting on Probe‐On Plus slides (Fisher Scientific).…”

Section: Introductionmentioning

confidence: 97%

“…The quantitative real-time PCR for CyHV-2 titration, tissue section preparation and ISH analysis were performed as described previously (Xu et al 2014). Peripheral blood smears were prepared as described by Masahiro et al (Yamasaki et al 2011). For ISH analysis, blood smears were formalin-fixed and paraffin-embedded before mounting on Probe-On Plus slides (Fisher Scientific).…”

Section: Introductionmentioning

confidence: 99%

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Detection of cyprinid herpesvirus 2 in peripheral blood cells of silver crucian carp, Carassius auratus gibelio (Bloch), suggests its potential in viral diagnosis

Wang

2015

Journal of Fish Diseases

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Epidemics caused by cyprinid herpesvirus 2 (CyHV-2) in domestic cyprinid species have been reported in both European and Asian countries. Although the mechanisms remain unknown, acute CyHV-2 infections generally result in high mortality, and the surviving carps become chronic carriers displaying no external clinical signs. In this study, in situ hybridization analysis showed that CyHV-2 tended to infect peripheral blood cells during either acute or chronic infections in silver crucian carp, Carassius auratus gibelio (Bloch). Laboratory challenge experiments coupled with real-time PCR quantification assays further indicated that steady-state levels of the viral genomic copy number in fish serum exhibited a typical 'one-step' growth curve post-viral challenge. Transcriptional expression of open reading frames (ORF) 121, which was selected due to its highest transcriptional levels in almost all tested tissues, was monitored to represent the replication kinetics of CyHV-2 in peripheral blood cells. Similar kinetic curve of active viral gene transcription in blood cells was obtained as that of serum viral load, indicating that CyHV-2 replicated in peripheral blood cells as well as in other well-characterized tissues. This study should pave the way for designing non-invasive and cost-effective serum diagnostic methods for quick detection of CyHV-2 infection.

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“…(Yamasaki et al . ). For ISH analysis, blood smears were formalin‐fixed and paraffin‐embedded before mounting on Probe‐On Plus slides (Fisher Scientific).…”

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Detection of cyprinid herpesvirus 2 in peripheral blood cells of silver crucian carp, Carassius auratus gibelio (Bloch), suggests its potential in viral diagnosis

Wang

2015

Journal of Fish Diseases

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“…In Beagle dogs experimentally infected with B. gibsoni , it was possible to detect the parasites with a probe matching a 516 bp DNA fragment of the heat-shock protein 70 (BgHsp70) gene via ISH. 73 FISH is available for the detection of other Babesia spp. ( B. duncani , B. microti ) in fresh human 61 and hamster blood smears.…”

Section: Discussionmentioning

confidence: 99%

Bovine Babesiosis Diagnosed in Formalin-Fixed, Paraffin-Embedded Tissues by Using In Situ Hybridization

Hülskötter¹,

Pfankuche²,

Dyck³

et al. 2020

Vet Pathol

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Bovine babesiosis, caused by Babesia divergens, is in general a rare disease in Europe. Nonetheless, local outbreaks can cause severe economic damage, and postmortem identification represents a diagnostic challenge. During a recent outbreak in May 2018 in northern Germany, 21 animals of a herd of 150 cattle died within 40 days having had clinical signs of fever and hemoglobinuria. Gross examination of 4 of the 21 deceased animals revealed a tick infestation, jaundice, and dark brown staining of urine and kidneys. Histologically, there were iron-positive deposits, hyperplasia of the red pulp of the spleen, and centrilobular necrosis of hepatocytes. In several locations, small basophilic granules suggestive of intraerythrocytic parasites were visible in hematoxylin-eosin- and Giemsa-stained sections. Peripheral blood smears from a living cow from the herd and polymerase chain reaction (PCR) of feeding ticks revealed B. divergens infection. In situ hybridization (ISH) was applied on formalin-fixed, paraffin-embedded (FFPE) tissue of the necropsied cattle to confirm babesiosis in these animals postmortem. Digoxigenin-labeled DNA probes were generated based on a specific nucleotide sequence for B. divergens, obtained by PCR and sequencing of DNA isolates from infected Ixodes ricinus ticks from deceased cattle. ISH using these probes allowed postmortem diagnosis of B. divergens infection in routinely fixed FFPE tissues.

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“…Thus, it is suggested that mtDNa should not be deeply involved in Da resistance in B. gibsoni. Babesia gibsoni is a blood protozoan of dogs and a causative pathogen of canine babesiosis [34]. It is difficult to eliminate this parasite from infected dogs, although a number of drugs, including diminazene aceturate (Da), clindamycin, metronidazole, pentamidine, and atovaquone combined with azithromycin, are used for treatment of the disease [9,16,22,29].…”

mentioning

confidence: 99%

Involvement of Mitochondrial Genes of <i>Babesia gibsoni</i> in Resistance to Diminazene Aceturate

Rajapakshage

Yamasaki

Hwang

et al. 2012

The Journal of Veterinary Medical Science

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aBsTracT. The stability of the characteristics of the diminazene aceturate (Da)-resistant B. gibsoni isolate was initially determined in vitro. Part of the Da-resistant B. gibsoni isolate was cultured without Da for 4 weeks, and then newly exposed to 200 ng/ml Da. as a result, this isolate could proliferate the same as the Da-resistant isolate, indicating that the characteristic of Da resistance was stable in the Daresistant isolate. additionally, the level of parasitemia in the Da-resistant isolate was comparatively lower than in the wild-type, suggesting that the proliferation potential of the Da-resistant isolate would be lower than that of the wild-type. subsequently, to investigate the involvement of mitochondrial DNa (mtDNa) in Da resistance in B. gibsoni, the nucleotide sequences and deduced amino acid sequences of mitochondrial genes such as COXI, COXIII, and CYTb genes of the Da-resistant isolate, were compared with those of the wild-type. as a result, these three genes were not altered in the Da-resistant B. gibsoni isolate. moreover, the transcription levels of COXI, COXIII, and CYTb genes were observed by semi-quantitative rT-Pcr. as a result, the gene transcription of those genes in the Da-resistant isolate was not significantly altered. These results indicated that DA did not affect mtDNA directly in DA-resistant B. gibsoni. Thus, it is suggested that mtDNa should not be deeply involved in Da resistance in B. gibsoni. Babesia gibsoni is a blood protozoan of dogs and a causative pathogen of canine babesiosis [34]. It is difficult to eliminate this parasite from infected dogs, although a number of drugs, including diminazene aceturate (Da), clindamycin, metronidazole, pentamidine, and atovaquone combined with azithromycin, are used for treatment of the disease [9,16,22,29]. in the treatment of canine babesiosis, possible relapses and the development of drug-resistant isolates are matters of concern.Da, an aromatic diamidine derivative, has been used as a first-line agent for the treatment of B. gibsoni infection in dogs [22]; however, Da cannot eliminate B. gibsoni from infected dogs, and relapses often occur [5,8,17,29]. in addition, a Da-resistant B. gibsoni isolate was previously developed in vitro in our laboratory [8]. However, the mechanism of action of Da on B. gibsoni has not been elucidated, and the mechanism of the development of Da resistance in B. gibsoni is still not known. To treat canine babesiosis with Da effectively, these problems have to be clarified. The DA-resistant B. gibsoni isolate developed by Hwang et al. was more resistant to pentamidine, clindamycin and doxycycline than the wild-type [8]. Furthermore, the transcription levels of the heat shock protein 70 (hsp70) gene in this isolate decreased during the development of the Da-resistant isolate, suggesting that the hsp70 gene is involved but does not play a major role in the development of the Da-resistant isolate. Thus, we have very little knowledge of the mechanism of the Da resistance of B. gibsoni. in Trypanosoma and Lei...

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Babesia gibsoni: Detection in blood smears and formalin-fixed, paraffin-embedded tissues using deoxyribonucleic acid in situ hybridization analysis

Cited by 6 publications

References 25 publications

Detection of cyprinid herpesvirus 2 in peripheral blood cells of silver crucian carp, Carassius auratus gibelio (Bloch), suggests its potential in viral diagnosis

Detection of cyprinid herpesvirus 2 in peripheral blood cells of silver crucian carp, Carassius auratus gibelio (Bloch), suggests its potential in viral diagnosis

Bovine Babesiosis Diagnosed in Formalin-Fixed, Paraffin-Embedded Tissues by Using In Situ Hybridization

Involvement of Mitochondrial Genes of <i>Babesia gibsoni</i> in Resistance to Diminazene Aceturate

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