Rapid diagnosis of lyme disease: Flagellin gene-based nested polymerase chain reaction for identification of causative Borrelia species

AbstractObjective: Each of Borrelia burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii has characteristic restriction sites in its flagellin gene. The authors focused on this gene and developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis for rapid diagnosis of Lyme disease.Methods: External and internal primer sets were designed for nested PCR to amplify an approximately 580 by fragment of the flagellin gene that includes species-specific restriction sites. DNA extracted from tissue samples of mice and humans were used as templates for PCR. The amplicons obtained were digested with HapII, HhaI, CelII HincII, or Ddel endonuclease.Results: In mice experimentally infected with each of B. burgdorferi sensu stricto, B. garinii, and B. afzelii, borrelial DNA was detected irrespective of differences in the causative species. However, RFLP of the amplicons was able to identify the species. Skin biopsy samples from 11 Japanese patients with erythema migrans were subjected to both PCR and culture tests. Borrelial infections were detected in seven cases (64%) by PCR and eight cases (73%) by culture. All PCR-positive samples were also positive by culture. The causative species in human infections was easily identified as B. garinii by RFLP analysis of the amplicons.Conclusion: The nested PCR-RFLP system appears to be an easy and reliable diagnostic tool for the detection and species identification of borreliae in human cutaneous biopsies

Report on the infectious disease group session

オガサワラカワラヒワ生息域外保全の専門家ワークショップにおいて、「感染症」が生存を脅かす可能性のある16の脅威のひとつに選出されたが、十分な議論時間を確保できなかった。そこで、専門的なグループセッションを実施して、オガサワラカワラヒワの保全を考える上で「感染症」をどう考えるべきか、情報の整理と議論を行った。小笠原諸島では鳥マラリアとポックスウイルスが確認されているが、感染症対策で野生個体群に対してできることは少ない。域外保全での検査と防除策の導入がベストな感染症的な知見の活かし方である。また、個体へのリスク管理上も、島外ではなく小笠原に繁殖保護施設をつくる等の意見が整理された。At the Ogasawara Greenfinch PHVA WS, "infectious diseases" was selected as one of the 16 threats. However, due to insufficient time for discussion, a specialized group session was held to organize and discuss information on "infectious diseases". Avian malaria and poxvirus have already been confirmed in the Ogasawara Islands. However, there are few measures that can be taken against wild populations. It is important to take inspection and control measures when implementing ex situ conservation. The creation of a local breeding shelter in Ogasawara is the right choice for managing the risk of infectious diseases.departmental bulletin pape

Prevalence of avian haemosporidia among injured wild birds in Tokyo and environs, Japan

Avian haemosporidia have been reported in various birds of Japan, which is part of the East Asian-Australian flyway and is an important stopover site for migratory birds potentially carrying new pathogens from other areas. We investigated the prevalence of avian malaria in injured wild birds, rescued in Tokyo and surrounding areas. We also evaluated the effects of migration by examining the prevalence of avian malaria for each migratory status. 475 birds of 80 species were sampled from four facilities. All samples were examined for haemosporidian infection via nested polymerase chain reaction (PCR) of the cytochrome b (cytb) gene. 100 birds (21.1%) of 43 species were PCR positive for avian haemosporidia. Prevalence in wintering birds, migratory breeders, and resident birds was 46.0%, 19.3%, 17.3% respectively. There was a bias in wintering birds due to Eurasian coot (Fulica atra) and Anseriformes. In wintering birds, lineages which are likely to be transmitted by Culiseta sp. in Northern Japan and lineages from resident species of Northern Japan or continental Asia were found, suggesting that wintering birds are mainly infected at their breeding sites. Meanwhile, there were numerous lineages found from resident and migratory breeders, suggesting that they are transmitted in Japan, some possibly unique to Japan. Although there are limits in studying rescued birds, rehabilitation facilities make sampling of difficult-to-catch migratory species possible and also allow for long-term monitoring within areas. Keywords: Avian haemosporidia, Japan, Rescued wild birds, Migratory birds, Parasite diversity, Cytochrome

Complete mitochondrial genome of a subspecies of the great cormorant, Phalacrocorax carbo hanedae (Kuroda, 1925) (Suliformes: Phalacrocoracidae)

We determined the complete mitochondrial DNA sequence of a subspecies of the great cormorant, Phalacrocorax carbo hanedae (Kuroda, 1925) using long PCR and primer walking methods. The mitochondrial genome was 19,020 bp in length and contained 13 protein-coding genes (PCGs), two ribosomal RNA genes, 22 transfer RNA genes, and two control regions. It is basically consistent with the characteristics of the mitochondrial genomes of other Suliformes species. Phylogenetic analysis using 12 species of Suliformes based on the sequences of 13 concatenated protein-coding genes confirmed the monophyly of P. carbo ssp

Complete mitochondrial genome of the Japanese Cormorant Phalacrocorax capillatus (Temminck & Schlegel, 1850) (Suliformes: Phalacrocoracidae)

The complete sequencing of mitochondrial DNA of the Japanese Cormorant Phalacrocorax capillatus was performed using long PCR and primer walking methods. The assembled genome was 19,105 bp in length. It contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and two control regions. The phylogenetic analysis using the obtained sequence showed that P. capillatus is closest to P. carbo