PMID- 33526178
OWN - NLM
STAT- MEDLINE
DCOM- 20210421
LR  - 20220107
IS  - 1878-1470 (Electronic)
IS  - 1568-9883 (Print)
IS  - 1568-9883 (Linking)
VI  - 101
DP  - 2021 Jan
TI  - Development of fluorescence in situ hybridization (FISH) probes to detect and 
      enumerate Gambierdiscus species.
PG  - 101914
LID - S1568-9883(20)30193-1 [pii]
LID - 10.1016/j.hal.2020.101914 [doi]
AB  - Ciguatera poisoning (CP) is a syndrome caused by the bioaccumulation of 
      lipophilic ciguatoxins in coral reef fish and invertebrates, and their subsequent 
      consumption by humans. These phycotoxins are produced by Gambierdiscus spp., 
      tropical epiphytic dinoflagellates that live on a variety of macrophytes, as well 
      as on dead corals and sand. Recent taxonomic studies have identified novel 
      diversity within the Gambierdiscus genus, with at least 18 species and several 
      sub-groups now identified, many of which co-occur and differ significantly in 
      toxicity. The ability to accurately and quickly distinguish Gambierdiscus species 
      in field samples and determine community composition and abundance is central to 
      assessing CP risk, yet most Gambierdiscus species are indistinguishable using 
      light microscopy, and other enumeration methods are semi-quantitative. In order 
      to investigate the spatial and temporal dynamics of Gambierdiscus species and 
      community toxicity, new tools for species identification and enumeration in field 
      samples are needed. Here, fluorescence in situ hybridization (FISH) probes were 
      designed for seven species commonly found in the Caribbean Sea and Pacific Ocean, 
      permitting their enumeration in field samples using epifluorescence microscopy. 
      This technique enables the assessment of community composition and accurate 
      determination of cell abundances of individual species. Molecular probes 
      detecting G. australes, G. belizeanus, G. caribaeus, G. carolinianus, G. 
      carpenteri, and the G. silvae/G. polynesiensis clade were designed using 
      alignments of large subunit ribosomal RNA (rRNA) sequences. These probes were 
      tested for specificity and cross-reactivity through experiments in which field 
      samples were spiked with known concentrations of Gambierdiscus cultures, and 
      analyzed to confirm that Gambierdiscus can be successfully detected and 
      enumerated by FISH in the presence of detritus and other organisms. These probes 
      were then used to characterize Gambierdiscus community structure in field samples 
      collected from the Florida Keys and Hawai'i, USA. The probes revealed the 
      co-occurrence of multiple species at each location. Time-series FISH analyses of 
      samples collected from the Florida Keys quantified seasonal shifts in community 
      composition as well as fluctuations in overall Gambierdiscus cell abundance. 
      Application of species-specific FISH probes provides a powerful new tool to those 
      seeking to target individual Gambierdiscus species, including significant 
      toxin-producers, in field populations. Moving forward, analysis of Gambierdiscus 
      community composition across multiple environments and over time will also allow 
      species dynamics to be linked to environmental parameters, improving our ability 
      to understand and manage the current and changing risks of CP worldwide.
CI  - Copyright (c) 2020. Published by Elsevier B.V.
FAU - Pitz, Kathleen J
AU  - Pitz KJ
AD  - Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, 
      USA.
FAU - Richlen, Mindy L
AU  - Richlen ML
AD  - Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, 
      USA. Electronic address: mrichlen@whoi.edu.
FAU - Fachon, Evangeline
AU  - Fachon E
AD  - Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, 
      USA.
FAU - Smith, Tyler B
AU  - Smith TB
AD  - Center for Marine and Environmental Studies, University of the Virgin Islands, St 
      Thomas, U.S. Virgin Islands 00802, USA.
FAU - Parsons, Michael L
AU  - Parsons ML
AD  - Coastal Watershed Institute, Florida Gulf Coast University, Fort Myers, FL 33965, 
      USA.
FAU - Anderson, Donald M
AU  - Anderson DM
AD  - Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, 
      USA.
LA  - eng
GR  - P01 ES028949/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20210106
PL  - Netherlands
TA  - Harmful Algae
JT  - Harmful algae
JID - 101128968
SB  - IM
MH  - Animals
MH  - Caribbean Region
MH  - *Dinoflagellida/genetics
MH  - Florida
MH  - In Situ Hybridization, Fluorescence
MH  - Pacific Ocean
PMC - PMC8016406
MID - NIHMS1660845
OTO - NOTNLM
OT  - Ciguatera
OT  - Ciguatoxin
OT  - Fluorescence in situ hybridization
OT  - Gambierdiscus
OT  - Harmful algal bloom
EDAT- 2021/02/03 06:00
MHDA- 2021/04/22 06:00
PMCR- 2022/01/06
CRDT- 2021/02/02 05:44
PHST- 2019/07/01 00:00 [received]
PHST- 2020/09/16 00:00 [revised]
PHST- 2020/10/05 00:00 [accepted]
PHST- 2021/02/02 05:44 [entrez]
PHST- 2021/02/03 06:00 [pubmed]
PHST- 2021/04/22 06:00 [medline]
PHST- 2022/01/06 00:00 [pmc-release]
AID - S1568-9883(20)30193-1 [pii]
AID - 10.1016/j.hal.2020.101914 [doi]
PST - ppublish
SO  - Harmful Algae. 2021 Jan;101:101914. doi: 10.1016/j.hal.2020.101914. Epub 2021 Jan 
      6.