PMID- 38462166
OWN - NLM
STAT- Publisher
LR  - 20240312
IS  - 1096-0805 (Electronic)
IS  - 0022-2011 (Linking)
VI  - 204
DP  - 2024 Mar 9
TI  - Overcoming research challenges: In vitro cultivation of Ameson portunus (Phylum 
      Microsporidia).
PG  - 108091
LID - S0022-2011(24)00034-X [pii]
LID - 10.1016/j.jip.2024.108091 [doi]
AB  - Ameson portunus is an intracellular pathogen that infects marine crabs Portunus 
      trituberculatus and Scylla paramamosain, causing significant economic losses. 
      However, research into this important parasite has been limited due to the 
      absence of an in vitro culture system. To address this challenge, we developed an 
      in vitro cultivation model of A. portunus using RK13 cell line in this study. The 
      fluorescent labeling assay indicated a high infection rate ( approximately 60 %) on the first 
      day post-infection and quantitative PCR (qPCR) detection demonstrated successful 
      infection as early as six hours post-inoculation. Fluorescence in situ 
      hybridization (FISH) and qPCR were used for the detection of A. portunus infected 
      cells. The FISH probe we designed allowed detection of A. portunus in infected 
      cells and qPCR assay provided accurate quantification of A. portunus in the 
      samples. Transmission electron microscopy (TEM) images revealed that A. portunus 
      could complete its entire life cycle and produce mature spores in RK13 cells. 
      Additionally, we have identified novel life cycle characteristics during the 
      development of A. portunus in RK 13 cells using TEM. These findings contribute to 
      our understanding of new life cycle pathways of A. portunus. The establishment of 
      an in vitro culture model for A. portunus is critical as it provides a valuable 
      tool for understanding the molecular and immunological events that occur during 
      infection. Furthermore, it will facilitate the development of effective treatment 
      strategies for this intracellular pathogen.
CI  - Copyright (c) 2024. Published by Elsevier Inc.
FAU - Fu, Ming
AU  - Fu M
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China; Marine College, 
      Shandong University, Weihai 264209, China.
FAU - Qu, Hongnan
AU  - Qu H
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Wang, Yongliang
AU  - Wang Y
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Guan, Jingyu
AU  - Guan J
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Xia, Tian
AU  - Xia T
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Zheng, Kai
AU  - Zheng K
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Tang, Liyuan
AU  - Tang L
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Zhou, Chunxue
AU  - Zhou C
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Zhou, Huaiyu
AU  - Zhou H
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China.
FAU - Cong, Wei
AU  - Cong W
AD  - Marine College, Shandong University, Weihai 264209, China. Electronic address: 
      messicw@163.com.
FAU - Zhang, Jinyong
AU  - Zhang J
AD  - School of Marine Science and Engineering, Qingdao Agricultural University, 
      Qingdao 266109, China. Electronic address: zhangjy@ihb.ac.cn.
FAU - Han, Bing
AU  - Han B
AD  - Department of Pathogenic Biology, School of Basic Medical Sciences, Cheeloo 
      College of Medicine, Shandong University, Jinan 250012, China. Electronic 
      address: bing.han@sdu.edu.cn.
LA  - eng
PT  - Journal Article
DEP - 20240309
PL  - United States
TA  - J Invertebr Pathol
JT  - Journal of invertebrate pathology
JID - 0014067
SB  - IM
OTO - NOTNLM
OT  - In vitro model
OT  - Intracellular pathogen
OT  - Marine crabs
OT  - Novel life cycle
OT  - RK13 cells
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2024/03/11 00:42
MHDA- 2024/03/11 00:42
CRDT- 2024/03/10 20:18
PHST- 2023/08/24 00:00 [received]
PHST- 2023/12/03 00:00 [revised]
PHST- 2024/03/06 00:00 [accepted]
PHST- 2024/03/11 00:42 [pubmed]
PHST- 2024/03/11 00:42 [medline]
PHST- 2024/03/10 20:18 [entrez]
AID - S0022-2011(24)00034-X [pii]
AID - 10.1016/j.jip.2024.108091 [doi]
PST - aheadofprint
SO  - J Invertebr Pathol. 2024 Mar 9;204:108091. doi: 10.1016/j.jip.2024.108091.