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The Aquaculture and Biology of 'Opihi 'Alinalina (Cellana Sandwicensis) a Dissertation Submitted to the Graduate Division Of THE AQUACULTURE AND BIOLOGY OF ‘OPIHI ‘ALINALINA (CELLANA SANDWICENSIS) A DISSERTATION SUBMITTED TO THE GRADUATE DIVISION OF THE UNIVERSITY OF HAWAI’I AT MANOA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN MOLECULAR BIOSCIENCES AND BIOENGINEERING DECEMBER 2019 By Anthony Mau Dissertation Committee: Jon-Paul Bingham (Chairperson) Rajesh Jha Cheng-Sheng Lee Birendra Mishra Kacie Ho Keywords: ecology, life-history, nutrition, ontogeny, reproduction, settlement Copyrights Copyright in the Dissertation held by the Author. All rights reserved. © 2019, Anthony B. Mau ii Dedication This dissertation is dedicated to my family, friends, and colleagues whom have inspired me to be a change-maker in our community. I specifically want to dedicate the completion of graduate school to my grandmother Laura, my mom Patricia, and my father Barry – whose unconditional love and financial support made it possible to attend Institutions like Punahou School, University of San Diego, and University of Hawai’i. “To whom much has been given, much will be required.” (Luke 12:48) I also dedicate this work to all of my heroes/best friends: Steven Yee (co-founder of Kupu Place LLC), William Gar Ho Hiroshi Lee M.D., Richard Charles Benner C.P.A., Brendan Big-Country Saunders, Luke O’connor, the Mission Beach crew, and the Semester-At-Sea crew; And of course, to my partner, Kira Miyuki Fox. Ke aloha! iii Acknowledgements I would like to formally acknowledge The Center for Tropical and Subtropical Aquaculture, The University of Hawai’i, The Hawaiian Malacological Society, and Dr. Steven Chun for funding this work. I want to mahalo everyone who has made this work possible from the bottom of my heart: Mahalo nui to my Committee Chairperson, Dr. Jon-Paul Bingham, who offered me a scholarly space to the conduct ‘opihi research in the Bingham Aquatic Laboratory at University of Hawai’i at Manoa. Under Dr. Bingham’s guidance, and during the most intense, enduring, and pivotal point of my career, I was able to think and work freely in this positive space. I was able to build a good deal of professional skills while we managed to write and fund multiple research grants, publish manuscripts, and mentor undergraduate students. And along with all of this, I will forever cherish both the banter and deep conversations we’ve shared. Mahalo nui to Dr. Rajesh Jha for guiding my technical writing, advising on experimental design, and teaching on aspects of Animal Nutrition and Nutritional Biochemistry; and for giving me a second home at the University of Hawai’i. Mahalo nui to Dr. Cheng-Sheng Lee for thoughtful conversations about aquaculture. You’ve played an instrumental role in getting me to attend graduate school, and have given me a platform to build my career on. I look up to you for your commitment to the aquaculture industry and local food production. iv Mahalo nui to Dr. Birendra Mishra for sharing your expertise on hormone regulation and endocrinology; and for opening your laboratory for histology work. Mahalo nui to Dr. Amy Moran for guidance on invertebrate biology and reproduction. Mahalo nui to Dr. Harry Ako for conversations on Biotechnology, career choices, and opportunities after Graduate School. I hope this work makes you proud. Mahalo nui to Dr. Erik Franklin for collaborating on the growth-modeling portion of this work. I’m elated to have met another surfer/researcher in the world of fisheries Mahalo nui to Dave Cohen at Anuenue Fisheries Research Center for supporting the next- generation of aquaculture in Hawai’i, and for always being a phone-call-away, especially when I had technical issues. Mahalo nui to Dr. Michael Hadfield for assistance with seeding settlement plates with biofilm, and for sharing your expertise in this field. Mahalo nui to my Undergraduate Students (future Masters/PhDs) for your kokua and collaboration: Ms. Angelica Valdez, Ms. Amihan Camson, Ms. Shelby Roberson, Mr. Philippe Nicodemus, Mr. Brandon Day, and Mr. John Paul Arios. v Mahalo nui to fellow ‘opihi researchers, Pelika Andrade (Sea Grant) and Dr. Randy Kosaki (NOAA), for the opportunity to contribute to ‘opihi conservation at Papahanaumokuakea Marine National Monument – a life-changing experience to say the least. Mahalo nui to Kehau Springer for sharing your home and connecting me with the ‘Opihi Partnership. And lastly, mahalo nui to Kim Morishige for your collaboration, leadership, and friendship during our most formative years! vi Abstract The present paradigm in global seafood production suggests that the market demands farm-raised seafood over wild-caught seafood. Whether this is due to a shift in market preference or simply an overall increase in farm production, there is an overwhelming amount of empirical evidence for diminishing return of wild stocks. For this reason, development of high-value mollusc species remains a top priority for the aquaculture industry – the ultimate goal being to improve food- security and reduce harvesting pressure on wild fisheries. For selection of new aquaculture species, it’s critical to understand the life-history to effectively engineer infrastructure, establish technical protocols, and optimize nutrition for production. In Hawai’i, ‘opihi ‘alinalina or yellowfoot limpet (Cellana sandwicensis), a prized mollusc shellfish and significant biocultural resource, has been a prospect aquaculture species for many years; and recently, there has been progress made towards closing the life cycle of C. sandwicensis in captivity. In this dissertation, the primary research objectives were to i) develop a suitable formulated feed by examining the effect of protein to energy ratio on growth performance of adult yellowfoot limpets in aquaculture; ii) establish suitable methodologies for controlled spawning and larval rearing to settlement; iii) examine the effects of a novel synthetic GnRH- like peptide hormone on limpet reproduction utilizing proteomics and solid-phase-peptide- synthesis; and iv) define the growth of yellowfoot limpets utilizing sclerochonology and stable isotope analysis by secondary ion mass spectrometry. The current advances of aquaculture technology and biology of C. sandwicensis includes: the fabrication and implementation of state-of-the-art recirculating aquaculture systems for broodstock, settlement, and grow-out; testing of a suitable formulated feed that meets nutritional vii requirements of the species; development of natural and hormone-controlled spawning techniques using a novel limpet GnRH-like peptide; establishment of methodology for larval rearing to settlement; and the first discoveries of key life-history traits. This research collective closes the present gap-in-knowledge pertaining to C. sandwicensis in aquaculture, and its biology – development, growth, and reproduction. Future research and development will focus on the settlement of limpets and the underlying mechanisms influencing survival in aquaculture, as well as to support ‘opihi fisheries management through understanding the life-history of C. sandwicensis. viii List of Publications Published Manuscripts: Chapter 1 Mau A, Jha R (2018) Aquaculture of two commercially important molluscs (abalone and limpet): existing knowledge and future prospects. Reviews in Aquaculture, 10, 611-625. Chapter 2: Mau A, Jha R (2018) Effects of dietary protein to energy ratios on growth performance of yellowfoot limpet (Cellana sandwicensis Pease, 1861). Aquaculture Reports, 10, 17-22. Chapter 3: Mau A, Fox K, Bingham JP (2017) The Reported Occurrence of Hermaphroditism in the Yellowfoot Limpet (Cellana sandwicensis Pease, 1981). Annals of Aquaculture Research, 4, 1045. Chapter 4: Mau A, Bingham JP, Soller F, Jha R (2018) Maturation, spawning, and larval development in captive yellowfoot limpets (Cellana sandwicensis Pease, 1861). Invertebrate Reproduction and Development, 62, 239-247. Submitted Manuscripts: Chapter 5: Mau A, Camson A, Bingham JP (2019) Gonadotropin Releasing Hormone-like peptide is not an acute spawn-inducer for Prosobranch Limpets (Cellana spp.). General and Comparative Endocrinology. Chapter 6: Mau A, Franklin EC, Nagashima K, Huss GA, Valdez AR, Nicodemus PN, Bingham JP (2019) Near-daily reconstruction of tropical intertidal SST from limpet shells to infer their growth rates. Nature Ecology & Evolution. ix Conference Proceedings: Mau A, Franklin EC, Nagashima K, Huss GA, Valdez AR, Nicodemus PN, Bingham JP (2020) Near-daily reconstruction of Tropical intertidal sea-surface temperature from limpet shells to infer their growth rates. Poster Presentation, Aquaculture America 2020 Conference and Exposition, Honolulu Convention Center, Honolulu, Hawai’i Mau A, Arios J, Valdez AR, Nicodemus P, Jha R, Bingham JP (2018) Chemical synthesis of a novel peptide oGnRH for pheromone stimulation of spawning in ‘opihi. Awarded CTAHR 30th annual Best Ph.D. Poster Presentation, 30th Annual CTAHR/COE SRS Symposium, University of Hawai’i at Manoa Mau A, Bingham JP (2017) Captive Maturation, Spawning, and Larval Rearing of ‘Opihi Cellana sandwicensis. Awarded Best Graduate Poster Presentation, 42nd Annual Albert L. Tester Symposium, Biology Department, University of Hawai’i at Manoa Mau A, Jha R (2017) The Protein to Energy Ratio Requirements of ‘Opihi Cellana sandwicensis. Awarded CTAHR Award of Merit for Oral Presentation, 29th Annual CTAHR/COE SRS Symposium, University of Hawai’i at Manoa x Table of Contents Page Dedication ....................................................................................................................
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