title: Development and validation of a competitive enzyme-linked immunosorbent assay for the measurement of total plasma immunoglobulins in healthy loggerhead sea (Caretta caretta) and green turtles (Chelonia mydas).
abstract: The quantification of circulating plasma immunoglobulins represents a valuable diagnostic tool in human and veterinary immunology, although its application is very limited in reptile medicine to date. The objectives of our study were the development and standardization of a competitive enzyme-linked immunosorbent assay (cELISA) for the measurement of total plasma immunoglobulins (Igs; both IgM and IgY) in loggerhead sea turtles (LST; Caretta caretta; n = 254) and green turtles (GT; Chelonia mydas; n = 111), the establishment of reference intervals for Ig for both species, and the examination of associations between Ig and total protein (TP), condition index, and water temperature. The cELISA for Ig was successfully developed and optimized. Reference intervals for Ig were 0.38-0.94 g/dL in LST (median: 0.59 g/dL; range: 0.16-2.15 g/dL) and 0.40-0.85 g/dL in GT (median: 0.58 g/dL; range: 0.18-1.80 g/dL). In LST, there were positive linear relationships of Ig with TP, and TP with Ig and condition index, and a negative relationship of Ig with condition index. The positive linear relationships of Ig with TP, and TP with Ig were also identified in GT. These positive associations of Ig and TP were expected, as Ig represents fractions of TP, and TP reportedly increases with straight carapace length and weight. The negative association of Ig with condition index may indicate potential biological variations. The cELISA and reference intervals for total Ig of LST and GT presented herein have the potential to be useful as a diagnostic and research tool for sea turtle immunology.
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Kaplan
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi) stacyn@ufl.edu.
authors.forename: Nicole I
authors.initials: NI
authors.lastname: Stacy
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Elliott
authors.initials: E
authors.lastname: Jacobson
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Carolina R
authors.initials: CR
authors.lastname: Le-Bert
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Hendrik H
authors.initials: HH
authors.lastname: Nollens
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Francesco C
authors.initials: FC
authors.lastname: Origgi
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Linda G
authors.initials: LG
authors.lastname: Green
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Shadi
authors.initials: S
authors.lastname: Bootorabi
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Alan
authors.initials: A
authors.lastname: Bolten
authors.affiliation: Departments of Small Animal Clinical Sciences (Kaplan, Jacobson, Hernandez), University of Florida, Gainesville, FLLarge Animal Clinical Sciences (Stacy, Le-Bert, Nollens, Hernandez), University of Florida, Gainesville, FLCollege of Veterinary Medicine, and the Hybridoma Core Laboratory, Interdisciplinary Center for Biotechnology Research (Green, Bootorabi), University of Florida, Gainesville, FLArchie Carr Center for Sea Turtle Research and Department of Biology (Bolten), University of Florida, Gainesville, FLNational Marine Mammal Foundation, San Diego, CA (Le-Bert)SeaWorld San Diego, San Diego, CA (Nollens)Centre for Fish and Wildlife Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland (Origgi).
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Hernandez
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D007136:Immunoglobulins / Q000097:blood*; D012016:Reference Values; D014426:Turtles / Q000097:blood*
chemical_list: D007136:Immunoglobulins
keywords: Enzyme-linked immunosorbent assay; immunoglobulin; sea turtles; serology