Establishing a Diagnostic Technique for Coelomocentesis in the Long-Spined Sea Urchin Diadema antillarum.

Coelomic fluid aspiration has been utilized in echinoderms in research and clinical settings. Detailed procedural descriptions for coelomic fluid sampling in sea urchins (class Echinoidea) are lacking, and samples are prone to contamination. The objectives of this study were to (1) standardize a technique for coelomic fluid collection in long-spined sea urchin Diadema antillarum that optimizes the diagnostic quality of the sample utilizing diagnostic imaging, (2) identify coelomic fluid bacterial isolates (using Biolog GEN III MicroLog and 16s rDNA sequencing), and (3) compare positive cultures to animal weight, holding time prior to sampling, water temperature, and gross fluid appearance. Seventy Diadema antillarum from the Florida Keys collected in two groups (March and September 2015) were utilized. Positive cultures for bacterial contamination were identified in 5% and 44%, respectively, of animals in the sampling groups. Vibrio spp. was the predominant genus identified. Positive cultures were more frequent in the group with smaller-sized animals, increased holding times, and elevated water temperatures. Deviation from clear-pink gross coelomic fluid appearance did not reliably predict bacterial contamination. A standardized technique for coelomocentesis was defined. The use of the proposed coelomocentesis methodology may facilitate improved health evaluations of sea urchins and may be applicable to research, conservation efforts, and disease investigations.

[1]  C. Morrow,et al.  Ocean warming alters predicted microbiome functionality in a common sea urchin , 2018, Proceedings of the Royal Society B: Biological Sciences.

[2]  G. Beck,et al.  Comparison of phagocytosis in three Caribbean Sea urchins , 2018, Developmental and comparative immunology.

[3]  C. Harms,et al.  TRICAINE METHANESULFONATE (MS-222) SEDATION AND ANESTHESIA IN THE PURPLE-SPINED SEA URCHIN (ARBACIA PUNCTULATA) , 2016, Journal of Zoo and Wildlife Medicine.

[4]  M. Bulling,et al.  New disease outbreak affects two dominant sea urchin species associated with Australian temperate reefs , 2016 .

[5]  H. Lessios.  The Great Diadema antillarum Die-Off: 30 Years Later. , 2016, Annual review of marine science.

[6]  C. López,et al.  Sea urchin Diadema africanum mass mortality in the subtropical eastern Atlantic: role of waterborne bacteria in a warming ocean , 2014 .

[7]  C. Faber,et al.  Systematic comparison and reconstruction of sea urchin (Echinoidea) internal anatomy: a novel approach using magnetic resonance imaging , 2008, BMC Biology.

[8]  T. Hughes.  Catastrophes, Phase Shifts, and Large-Scale Degradation of a Caribbean Coral Reef , 1994, Science.

[9]  H. Lessios,et al.  MASS MORTALITY OF DIADEMA ANTILLARUM IN THE CARIBBEAN: What Have We Learned? , 1988 .

[10]  R. Scheibling,et al.  Experimental infection of the echinoid strongylocentrotus droebachiensis with Paramoeba invadens: quantitative changes in the coelomic fluid , 1988 .

[11]  J. Bauer,et al.  Isolation of bacteria pathogenic for the sea urchin Diadema antillarum (Echinodermata: Echinoidea) , 1987 .

[12]  T. Hughes,et al.  Herbivory on coral reefs: community structure following mass mortalities of sea urchins , 1987 .

[13]  Kw Gilles,et al.  Disease in sea urchin Strongylocentrotus purpuratus: experimental infection and bacterial virulence , 1985 .

[14]  T. Hughes,et al.  Mass mortality of the echinoid Diadema antillarum Philippi in Jamaica , 1985 .

[15]  D. R. Robertson,et al.  Spread of Diadema Mass Mortality Through the Caribbean , 1984, Science.

[16]  S. Unkles,et al.  Bactericidal activity of coelomic fluid from the sea urchin , 1978 .

[17]  S. Unkles.  Bacterial flora of the sea urchin Echinus esculentus , 1977, Applied and environmental microbiology.