Sequence and phylogenetic analysis of the gp200 protein of Ehrlichia canis from dogs in Taiwan

Ehrlichia (E.) canis is a Gram-negative obligate intracellular bacterium responsible for canine monocytic ehrlichiosis. Currently, the genetic diversity of E. canis strains worldwide is poorly defined. In the present study, sequence analysis of the nearly full-length 16S rDNA (1,620 bp) and the complete coding region (4,269 bp) of the gp200 gene, which encodes the largest major immunoreactive protein in E. canis, from 17 Taiwanese samples was conducted. The resultant 16S rDNA sequences were found to be identical to each other and have very high homology (99.4~100%) with previously reported E. canis sequences. Additionally, phylogenetic analysis of gp200 demonstrated that the E. canis Taiwanese genotype was genetically distinct from other reported isolates obtained from the United States, Brazil, and Israel, and that it formed a separate clade. Remarkable variations unique to the Taiwanese genotype were found throughout the deduced amino acid sequence of gp200, including 15 substitutions occurring in two of five known species-specific epitopes. The gp200 amino acid sequences of the Taiwanese genotype bore 94.4~94.6 identities with those of the isolates from the United States and Brazil, and 93.7% homology with that of the Israeli isolate. Taken together, these results suggest that the Taiwanese genotype represents a novel strain of E. canis that has not yet been characterized

Association of anticardiolipin, antiphosphatidylserine, anti-β2 glycoprotein I, and antiphosphatidylcholine autoantibodies with canine immune thrombocytopenia

β2GPI expression and identification. (PDF 159 kb

Molecular+Characterization+on+18S+rRNA+Gene+of+Hepatozoon+canis+from+Suspected+Clinical+Infections+in+Taiwan+Dog+Cases

The aim of this study was to investigate the species of Hepatozoon spp. in dogs in Taiwan. Blood samples were collected from 714 dogs and DNA was extracted and analyzed by the polymerase chain reaction. Of the 714 animals, 15 (2.1%) cases were polymerase chain reaction-positive with Hepatozoon-genus specific primers. By sequence analysis, the partial sequences (666 bp) of the 18S rRNA gene among specified 15 samples with Hepatozoon spp. were shown mutually identical and with 100% similarity to other sequences of H. canis isolates including two Spain cases respectively from Old World red fox Vulpes vulpes (AY731062) and dog (AY461378), one Venezuela case from dog (DQ439543), and one USA case from dog (AF176835). With the phylogenetic tree in this report, the Taiwan cases with H. canis DNA isolates were shown at closest relevance to the cases with Vennezuela 1, Spain 2, Spain 3, USA and Australia strains. To our knowledge, this clinical case report represents the initiative molecular characterization on the 18S rRNA gene sequences of H. canis DNA isolates within the dog population in Taiwan

A+Preliminary+Study+of+Using+the+PCR-DGGE+Assay+for+Detection+the+Autosomal+Dominant+Polycystic+Kidney+Disease+in+Domestic+Cats

貓之體染色體顯性多囊性腎臟病（ADPKD）被認為是貓最常見家族性遺傳性疾病，常發生在波絲貓或是有波絲貓血統的貓。貓PKD1等位基因第29外顯子（exon）中的第3284個核苷酸轉換（C→A），所導致之點突變，是造成AD-PKD原因之一。目前臨床上主要使用腎臟超音波造影檢測方式來診斷ADPKD，本研究首先從2009至2010年間至中興大學獸醫教學醫院就診的貓隻，共收集7個樣本，利用腎臟超音波造影檢測初步診斷得到AD-PKD之4樣本，進而利用PCR-DGGE檢測，結果亦可檢測出4個樣本有PKD1等位基因外顯子突變。由此研究結果顯示利用PCRDGGE鑑別AD-PKD突變株的靈敏度及準確度與腎臟超音波造影檢查結果一致，故在診斷AD-PKD上提供另外一種分子鑑定分析技術之選擇。The autosomal dominant polycystic kidney disease (AD-PKD) is a familial disease in cats and is characterized by the occurrence of fluid-filled cysts in the renal tissues. The cysts often lead to the kidney failure. The major cause of disease is a C→A point mutation at nt 3284 in the PKD1 gene. In clinical diagnosis, the renal ultrasounds are often used to diagnose AD-PKD. In this study, 7 blood samples were collected and analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method to detect single nucleotide polymorphisms (SNPs) in PKD1 gene. Among the 7 samples, 4 samples were preliminarily diagnosed with AD-PKD by renal ultrasounds. The 7 samples were further analyzed by PCRDGGE, and only 4 samples were diagnosed with AD-PKD. Therefore, the sensitivity and the accuracy of PCR-DGGE provides an alternative technique to distinguish AD-PKD mutants in cats

Sequence+and+Phylogenetic+Analyses+of+the+18S+rRNA+Gene+of+Babesia+canis+in+Dogs+in+Central+Taiwan

Canine babesiosis represents an important veterinary medical problem. The present study aimed at characterizing Babesia canis strains in naturally infected dogs in central Taiwan, using a combination of polymerase chain reaction (PCR) and sequence analysis of 18S rRNA gene. Genomic DNA was extracted from 36 canine blood samples and the target genes were separately amplified, sequenced, and aligned with corresponding sequences available in the GenBank database. All 18S rDNA sequences (455 bp) amplified from the ten Taiwanese isolates were identical to each other and had very high similarity (99.1-99.5%) with previously reported Babesia canis vogeli sequences. A phylogenetic analysis based on the 18S rDNA sequence demonstrated that the Taiwanese isolates were closely related to a strain previously identified from Japan. These results provided the first molecular evidence showing infection of dogs with B. canis vogeli from Taiwan.本研究主要針對台灣中部地區自然感染焦蟲症之犬隻，利用聚合酶連鎖反應及18S rRNA基因定序方法進行病原體檢測及其分子特性之探討。首先抽取36個犬血液樣本的基因體DNA，然後分別擴增標的基因之節段、定序並與GenBank資料庫中的相關序列進行核苷酸排比。所有自10個台灣分離株擴增出來的18S rDNA序列（455個鹼基對長）全皆相同，並且與先前其他國家或地區所發表的Babesia canis vogeli序列有非常高的相似性（99.1-99.5%）。根據18S rDNA序列所做的親緣分析結果顯示，台灣分離株和先前已確認的日本株相近。這些結果首度提供台灣犬隻感染B. canis vogeli的分子證據

台灣中部地區貓隻感染血巴東蟲之調查暨分子特性分析

The aim of this study was to survey Mycoplasma spp. infection in cats in central Taiwan, and to characterize Mycoplasma spp. by polymerase chain reaction (PCR). We collected 120 feline blood samples at National Chung-Hsing University Veterinary Medical Teaching Hospital and 5 veterinary hospitals from March to September 2011, and these samples were analyzed by PCR and blood examination. Of the 120 samples, 5 (4.2%) were Mycoplasma haemofelis (Mhf), 21 (17.5%) were Candidatus Mycoplasma haemominutum (CMhm), 2 (1.6%) cats were Candidatus Mycoplasma turicensis (CMt), and 3 (2.5%) were Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis. The main symptoms of Mhf-infected cats included anemia, anorexia, and depression; and those of CMhm-infected cats included anemia and depression; and those of CMt-infected cats included anemia. In addition, we speculated that male, mixed breed and less than six-year-old cats were the risk factors for CMhm infection. The further 16S rRNA gene sequence analysis showed that the sequences of 5 Mhf Taiwan strains shared 100% identity with USA strain, and the sequences of 21 CMhm Taiwan strains shared 99.4-99.8% identity with United Kingdom and Israel strains, and the sequences of 2 CMt Taiwan strains shared 100% identity with Switzerland and South Africa strains. Phylogenetic analysis showed that Mhf, CMhm and CMt Taiwan strains were grouped with the same species. The data indicate that the main Mycoplasma spp. pathogens in cats in central Taiwan are CMhm, followed by Mhf and CMt.本研究目的為利用聚合酶連鎖反應（Polymerase chain reaction, PCR）方法調查台灣中部地區貓隻感染之Mycoplasma病原體，且進一步進行分子特性分析。本研究於100年3月至9月期間收集120隻於國立中興大學獸醫教學醫院及5間獸醫診所就診之貓隻抗凝血血液進行PCR檢驗及血液學分析。結果顯示共5（4.2%）隻貓隻為Mycoplasma haemofelis（Mhf）陽性，21（17.5%）貓隻為Candidatus Mycoplasma haemominutum（CMhm）陽性及2（1.6 %）隻貓隻為Candidatus Mycoplasma turicensis（CMt）陽性，3（2.5%）隻貓隻共同感染Mhf 及CMhm。感染Mhf的貓隻主要症狀包括貧血（80%）、厭食（100%）及精神抑鬱（80%）；感染CMhm的貓隻主要症狀包括貧血（90.5%）及精神抑鬱（81%）；而感染CMt的貓隻主要症狀包括貧血（100%）。此外，本研究推測公貓、混種貓及年齡低於6歲貓隻為感染CMhm的可能相關危險因子。進一步進行16S rRNA基因分析顯示5個Mhf序列與美國株序列相似度為100%，21個CMhm序列與英國及以色列株相似度為99.4-99.8%，2個CMt序列則分別與瑞士及南非株之相似度為100%。親緣關係樹狀圖顯示Mhf、CMhm及CMt皆聚集於相同種的族群。由本研究結果顯示台灣中部地區貓隻CMhm感染率最高，再來是Mhf及CMt

Association of anticardiolipin, antiphosphatidylserine, anti-β2 glycoprotein I, and antiphosphatidylcholine autoantibodies with canine immune thrombocytopenia

In humans, the presence of antiphospholipid antibodies (aPL) is frequently found in immune thrombocytopenia. The present study investigated whether aPL and any aPL subtypes are associated with canine thrombocytopenia, in particular, immune-mediated thrombocytopenia (immune thrombocytopenia) that usually manifests with severe thrombocytopenia

Additional file 6: of Association of anticardiolipin, antiphosphatidylserine, anti-β2 glycoprotein I, and antiphosphatidylcholine autoantibodies with canine immune thrombocytopenia

Frequencies of aPL subtypes in all thrombocytopenic dogs. (PDF 47 kb