Barnacle biology before, during and after settlement and metamorphosis: a study of the interface.
Mobile barnacle cypris larvae settle and metamorphose, transitioning to sessile juveniles with morphology and growth similar to that of adults. Because biofilms exist on immersed surfaces on which they attach, barnacles must interact with bacteria during initial attachment and subsequent growth. The objective of this study was to characterize the developing interface of the barnacle and substratum during this key developmental transition to inform potential mechanisms that promote attachment. The interface was characterized using confocal microscopy and fluorescent dyes to identify morphological and chemical changes to the interface and the status of bacteria present as a function of barnacle developmental stage. Staining revealed patchy material containing proteins and nucleic acids, reactive oxygen species amidst developing cuticle, and changes in bacteria viability at the developing interface. We found that as barnacles metamorphose from the cyprid to juvenile stage, proteinaceous materials with the appearance of coagulated liquid were released into and remained at the interface. It stained positive for proteins, including phosphoprotein, as well as nucleic acids. Regions of the developing cuticle and the patchy material itself stained for reactive oxygen species. Bacteria were absent until the cyprid was firmly attached, but populations died as barnacle development progressed. The oxidative environment may contribute to the cytotoxicity observed for bacteria and has the potential for oxidative crosslinking of cuticle and proteinaceous materials at the interface.
Kewalo Marine Laboratory, Pacific Biosciences Research Center, University of Hawaii, 41 Ahui St, Honolulu, HI 96813, USA essock@hawaii.edu.
Tara
0000-0003-4159-6974
T
Essock-Burns
Okeanos Research Group, Department of Biological Sciences, 132 Long Hall, Clemson University, Clemson, SC 29634, USA.
Neeraj V
NV
Gohad
Duke University Marine Laboratory, Marine Science and Conservation, 135 Duke Marine Lab Road, Beaufort, NC 28516, USA.
Beatriz
B
Orihuela
Duke University Marine Laboratory, Marine Science and Conservation, 135 Duke Marine Lab Road, Beaufort, NC 28516, USA.
Andrew S
AS
Mount
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USA.
Christopher M
CM
Spillmann
Chemistry Division, Naval Research Laboratory, Washington, DC 20375, USA.
Kathryn J
KJ
Wahl
Duke University Marine Laboratory, Marine Science and Conservation, 135 Duke Marine Lab Road, Beaufort, NC 28516, USA.
Daniel
D
Rittschof
D000818:Animals; D060829:Arthropod Proteins / Q000378:metabolism; D018407:Bacterial Physiological Phenomena; D018441:Biofilms / Q000254:growth & development*; D005456:Fluorescent Dyes; D007814:Larva / Q000254:growth & development / Q000378:metabolism / Q000382:microbiology; D008675:Metamorphosis, Biological; D018613:Microscopy, Confocal; D009696:Nucleic Acids / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D001468:Thoracica / Q000254:growth & development* / Q000378:metabolism / Q000382:microbiology
D060829:Arthropod Proteins; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D017382:Reactive Oxygen Species
Antimicrobial; Attachment; Bacteria; Confocal; Fouling; Reactive oxygen species