PMID- 25167936
OWN - NLM
STAT- MEDLINE
DCOM- 20160405
LR  - 20200930
IS  - 1365-2761 (Electronic)
IS  - 0140-7775 (Linking)
VI  - 38
IP  - 9
DP  - 2015 Sep
TI  - Direct visualization of the novel pathogen, Spiroplasma eriocheiris, in the 
      freshwater crayfish Procambarus clarkii (Girard) using fluorescence in situ 
      hybridization.
PG  - 787-94
LID - 10.1111/jfd.12287 [doi]
AB  - Spiroplasma eriocheiris is the first spiroplasma strain known to be pathogenic to 
      freshwater crustaceans. It has caused considerable economic losses both in the 
      freshwater crayfish Procambarus clarkii (Girard) and in some other crustaceans. 
      The monitoring of the pathogen in crustacean populations and study of its 
      behaviour in the laboratory require the development of reliable diagnostic tools. 
      In this article, we improved microscopic identification of S. eriocheiris by 
      combining in situ hybridization with specific fluorescently labelled 
      oligonucleotide probes. The established fluorescence in situ hybridization (FISH) 
      allowed simultaneous visualization, identification and localization of 
      S. eriocheiris in the tissues of diseased crayfish P. clarkii and exhibited low 
      background autofluorescence and ideal signal-to-noise ratio. With the advantages 
      of better tissue penetration, potentially more specific and stable, we designed 
      three species-specific oligonucleotide probes utilizing the sequences of 16S-23S 
      rRNA intergenic spacer regions (ISRs) of S. eriocheiris. Positive hybridization 
      signals were visualized in haemocytes and connective tissues of hepatopancreas, 
      cardiac muscle and gill from diseased crayfish. This unique distribution pattern 
      matched the pathological changes when diagnosed by H&E staining and indicated 
      that S. eriocheiris probably spread throughout the tissues in P. clarkii by 
      hemokinesis. This assay will facilitate our understanding of the pathogenesis of 
      S. eriocheiris and enhance the early diagnosis of the novel pathogen.
CI  - (c) 2014 John Wiley & Sons Ltd.
FAU - Ding, Z F
AU  - Ding ZF
AD  - Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key 
      Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing 
      Normal University, Nanjing, China.
AD  - Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing, China.
FAU - Xia, S Y
AU  - Xia SY
AD  - College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 
      China.
FAU - Xue, H
AU  - Xue H
AD  - Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing, China.
FAU - Tang, J Q
AU  - Tang JQ
AD  - Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing, China.
FAU - Ren, Q
AU  - Ren Q
AD  - Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key 
      Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing 
      Normal University, Nanjing, China.
FAU - Gu, W
AU  - Gu W
AD  - Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key 
      Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing 
      Normal University, Nanjing, China.
FAU - Meng, Q G
AU  - Meng QG
AD  - Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key 
      Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing 
      Normal University, Nanjing, China.
FAU - Wang, W
AU  - Wang W
AD  - Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key 
      Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing 
      Normal University, Nanjing, China.
LA  - eng
SI  - GENBANK/AY549216
SI  - GENBANK/AY729936
SI  - GENBANK/AY736030
SI  - GENBANK/DQ004913
SI  - GENBANK/DQ917754
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140829
PL  - England
TA  - J Fish Dis
JT  - Journal of fish diseases
JID - 9881188
SB  - IM
MH  - Animals
MH  - Aquaculture/*methods
MH  - Astacoidea/*microbiology
MH  - Fresh Water
MH  - Hepatopancreas/microbiology/pathology
MH  - In Situ Hybridization, Fluorescence
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction
MH  - Spiroplasma/*cytology/genetics
OTO - NOTNLM
OT  - Procambarus clarkii
OT  - Spiroplasma eriocheiris
OT  - fluorescence in situ hybridization
OT  - pathogenesis
EDAT- 2014/08/30 06:00
MHDA- 2016/04/06 06:00
CRDT- 2014/08/30 06:00
PHST- 2014/05/13 00:00 [received]
PHST- 2014/06/11 00:00 [revised]
PHST- 2014/06/19 00:00 [accepted]
PHST- 2014/08/30 06:00 [entrez]
PHST- 2014/08/30 06:00 [pubmed]
PHST- 2016/04/06 06:00 [medline]
AID - 10.1111/jfd.12287 [doi]
PST - ppublish
SO  - J Fish Dis. 2015 Sep;38(9):787-94. doi: 10.1111/jfd.12287. Epub 2014 Aug 29.