The 2nd International Symposium on New Frontiers in Reef Coral Biotechnology (12 May 2023, Taiwan)

Abstract

For the second year in a row, the theme is “reef coral biotechnology”, specifically the interface between basic science and conservation. It has never been more important to attempt to leverage what we know about these beautiful, albeit highly imperiled and fragile, ecosystems towards conserving them. Our invited speakers’ areas of expertise span all levels of biological organization: from molecules within coral cells, to coral tissues, to entire coral colonies, and then up to reef-scale processes. Our goal is to promote communication not only among local Taiwanese marine biologists, but also those within Southeast Asia and farther afield; we especially encourage participation from early-career researchers, including Master’s students, PhD candidates, and post-doctoral researchers. It is our hope that the presentations (and the discussions that follow) will encourage collaboration. As importantly, we envision that the tools and approaches shared amongst us can be tapped into to expedite our collective efforts to better understand, manage, and conserve coral reefs.

Full text

Citation: Lin, C. The 2nd
International Symposium on New
Frontiers in Reef Coral Biotechnology
(12 May 2023, Taiwan). Appl. Sci.
2023,13, 7318. https://doi.org/
10.3390/app13127318
Academic Editor: Dibyendu Sarkar
Received: 15 June 2023
Accepted: 17 June 2023
Published: 20 June 2023
Copyright: © 2023 by the author.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
applied
sciences
Conference Report
The 2nd International Symposium on New Frontiers in Reef
Coral Biotechnology (12 May 2023, Taiwan)
Chiahsin Lin 1,2
1
Institute of Marine Biology, National Dong Hwa University, Pingtung 94450, Taiwan; chiahsin@nmmba.gov.tw
2Department of Planning and Research, National Museum of Marine Biology & Aquarium,
Pingtung 94450, Taiwan
Abstract:
For the second year in a row, the theme is “reef coral biotechnology”, specifically the
interface between basic science and conservation. It has never been more important to attempt to
leverage what we know about these beautiful, albeit highly imperiled and fragile, ecosystems towards
conserving them. Our invited speakers’ areas of expertise span all levels of biological organization:
from molecules within coral cells, to coral tissues, to entire coral colonies, and then up to reef-scale
processes. Our goal is to promote communication not only among local Taiwanese marine biologists,
but also those within Southeast Asia and farther afield; we especially encourage participation from
early-career researchers, including Master’s students, PhD candidates, and post-doctoral researchers.
It is our hope that the presentations (and the discussions that follow) will encourage collaboration.
As importantly, we envision that the tools and approaches shared amongst us can be tapped into to
expedite our collective efforts to better understand, manage, and conserve coral reefs.
Keywords:
artificial intelligence; coral restoration; endosymbiosis; health monitoring;
cryopreservation
;
husbandry
Genomic Signatures Underlying the Adaptability and Resilience of Scleractinian Corals
Cecilia Conaco *, Niño Posadas and Jeric
Da-Anoy----------------------------------------------------------------- 2
Data-Driven Marine Conservation in the Coral Triangle
Anderson B. Mayfield *---------------------------------------------- 2
Rehabilitating Reef Rehabilitation: Applying Scientific Knowledge to Improve Reef
Restoration Practice in Indonesia
Tries Razak *------------------------------------------------------------ 3
Importance of Lipid Droplets in Endosymbiotic Marine Dinoflagellate Associated
with Corals
Buntora Pasaribu *, Noir Primadona Purba, Alex Muhammad Akbar
Khan, Ibnu Faizal, Lantun Paradhita Dewanti, Mega Laksmini
Syamsuddin, Yudi Nurul Ihsan, Rita Rostika,
Syawaludin Harahap and Pei Luen Jiang-----------------------
3
Low-Technology In-Vitro Cultivation of Small Corals Fragments: A Model System for Clone
Production towards Developing Quality R & D
Alex P. Camaya *, Satoko Sekida and Kazuo Okuda--------- 4
Proteomic Analysis of Acropora cervicornis Exposed to Thermal Acclimation
Samantha Shaw *, Anderson Mayfield, Cliff Ross and
Matthew Gilg----------------------------------------------------------- 5
Heat Tolerance and Symbiodiniaceae Profiles of Acroporids in a Philippine Reef
John Bennedick Quijano *, Jake Ivan P. Baquiran and Cecilia
Conaco------------------------------------------------------------------- 5
The First Cryo-Repository for Coral Larvae: Safeguarding Corals for Future Generations
Kanokpron Loeslakwiboon, Wen-Chung Hsieh, Cheng-Liang Huang,
Sujune Tsai and Chiahsin Lin *------------------------------------- 5
Appl. Sci. 2023,13, 7318. https://doi.org/10.3390/app13127318 https://www.mdpi.com/journal/applsci

Appl. Sci. 2023,13, 7318 2 of 9
Coral Assessment in Selected Areas in the Verde Island Passage
Enriquo Martin C. Velasquez *, Miguel Enrique Ma. A. Azcuna, Jonel
M. Corral, Jade Symon A. Binay, Karlo L. Pintor, Rex M. Medina,
Katherine Sanchez-Escalona, Fhred Kiervin Montante and Jayvee
Ablaña Saco--------------------------------------------------------------
6
Differential Symbiodiniaceae Association with Acropora Humilis Coral While Rearing in
Hatchery Conditions
Suppakarn Jandang *, Voranop Viyakarn, Yuki Yoshioka, Chuya
Shinzato and Suchana Chavanich---------------------------------- 7
Coral Restoration Program in the Bicol Region, the Philippines: Experiences and Challenges
Antonino B. Mendoza *, Satoshi Kubota and Plutomeo M.
Nieves--------------------------------------------------------------------- 7
Spatiotemporal Lipid Profiling during Coral Embryogenesis
Federica Buttari, Sujune Tsai, Zhi-Hong Wen and Chiahsin
Lin *------------------------------------------------------------------------ 8
Benthic Community Structures with Varying Sedimentation and Water Quality Conditions in
Mabini, Batangas
Danilo A. Leyble Jr. *, Riza Maree D. C. Rapada, Ethel C. Wagas and
Milette U. Mendoza-Pascual----------------------------------------- 8
Characteristics of Black Pigment Released from the Octocoral Sinularia flexibilis
Lian-Di Lin, Yu-Chia Chang and Hsing-Hui Li *---------------- 9
1. Genomic Signatures Underlying the Adaptability and Resilience of
Scleractinian Corals
Cecilia Conaco *, Niño Posadas and Jeric Da-Anoy
Marine Science Institute, University of the Philippines, Diliman, Quezon City, Philippines
∗Correspondence: cconaco@msi.upd.edu.ph
Abstract
Corals today face increasingly challenging conditions, including rising ocean tem-
peratures brought about by climate change. To gain insight into how phylogenetically
divergent species may respond to thermal stress events, we compared the gene complement
of four coral species, Acropora digitifera,Favites colemani,Montipora digitata, and Seriatopora
caliendrum, that were previously shown to have different sensitivities to acute thermal
stress. The comparison of gene repertoires in the four scleractinian corals revealed an
extensive set of cnidarian stress response genes (SRGs). Antioxidant protein families and
chaperones were relatively more abundant in F. colemani and M. digitata; proteins linked to
immunological functions were enriched in S. caliendrum and A. digitifera; and extracellular
matrix components were abundant in S. caliendrum and F. colemani. The species that showed
greatest heat susceptibility, S. caliendrum, also exhibited drastic upregulation of SRGs under
acute thermal stress. This suggests that differences in SRGs, as well as the mechanisms
that control SRG expression response, contribute to the ability of corals to maintain stable
physiological functions and survive shifts in seawater temperature.
2. Data-Driven Marine Conservation in the Coral Triangle
Anderson B. Mayfield
International Coral Reef Society, Miami, FL, USA; anderson@coralreefdiagnostics.com
Abstract
Due to the plethora of threats to coral reef ecosystems, marine biologists have been
attempting to both document reef decline and devise conservation solutions that could
thwart coral extinction or partially restore ecosystem function. In most cases, local-scale
stressors are not adequately addressed prior to initiating these projects, not because re-
searchers do not appreciate their impact but because, in many cases, it is not possible to do
so on a timescale that is commensurate with coral rescue. What this means, though, is that
many coral reef conservation initiatives are doomed to fail from the start, notably many

Appl. Sci. 2023,13, 7318 3 of 9
restoration projects that seek to grow corals for later outplanting on the reef; if temperatures
continue to rise, and seawater quality continues to deteriorate, only those projects that
stress-harden, genetically modify, or, more generally, modulate the underlying biology
of the farmed corals have any hope of success. Ideally, a manager would collect some
rudimentary data on the habitat in question, then use statistical tools to determine the
conservation approach that has the highest probability of success due to what is known
about the habitat in question. This can be achieved with machine learning, a modeling
approach based on artificial intelligence (AI), though the marine biology field has generally
been slow to leverage breakthroughs in such “big data” analytics that have revolutionized
other disciplines, such as advertising. I discuss a “coral rescue” flow chart and show how
an AI could be trained to robustly project the conservation approach with the highest
probability of success (e.g., maximum coral cover, longest coral colony lifespan, etc.).
3. Rehabilitating Reef Rehabilitation: Applying Scientific Knowledge to Improve Reef
Restoration Practice in Indonesia
Tries Razak
National Research and Innovation Agency, Jakarta, Indonesia; tbrazak@gmail.com
Abstract
Indonesia’s coral reefs have been severely damaged by global and local stressors, and
a range of active restoration techniques are now used in attempts to rebuild degraded reefs.
However, it is difficult to summarize Indonesia’s restoration efforts due to a lack of consis-
tent reporting. Here, we first discuss Indonesia’s legal policy framework concerning reef
restoration. We then provide an extensive review of reef restoration projects in Indonesia,
documenting 533 records between 1990 and 2020. Most (73%) of these records come from
the past 10 years, and many (42%) are reported in online news articles. This review identi-
fied 120,483 units of artificial reef installed across Indonesia, along with 53,640 units of coral
transplantation (including both coral nurseries and direct out-planting onto reefs); in total,
965,992 fragments of hard coral have been planted across Indonesia. The most favoured
restoration materials are concrete (46%) and steel structures (24%). Projects are organized
by a diverse range of governmental, NGO, private and community-led organizations. This
review demonstrates that Indonesia’s policy has encouraged a diverse range of practitioners
to implement reef restoration, but projects are often not coordinated with wider networks
of restoration practitioners or scientists, and only 16% of the identified projects included
a post-installation monitoring framework. Incorporating clear objectives and long-term
monitoring programmes in project planning stages, while prioritizing knowledge exchange
and engagement with the international scientific community, will substantially improve
restoration outcomes in Indonesia. This will allow the country to fulfill its considerable
potential as a global leader in rebuilding damaged coral reefs.
4. Importance of Lipid Droplets in Endosymbiotic Marine Dinoflagellate Associated
with Corals
Buntora Pasaribu 1,2,*,Noir Primadona Purba 1, Alex Muhammad Akbar Khan 3,
Ibnu Faizal 1, Lantun Paradhita Dewanti 4, Mega Laksmini Syamsuddin 1,
Yudi Nurul Ihsan 1, Rita Rostika 4, Syawaludin Harahap 1and Pei Luen Jiang 5
1
Department of Marine Science, Faculty of Fisheries and Marine Science, Universitas
Padjadjaran, Bandung 40600, Indonesia
2Marine and Biogas Research Laboratory, Bandung 40600, Indonesia
3
Tropical Marine Fisheries Undergraduate Programme for Pangandaran Campus,
Faculty of Fisheries and Marine Science, Universitas Padjadjaran, Bandung 40600,
Indonesia
4
Fisheries Study Programme, Faculty of Fisheries and Marine Science, Universitas
Padjadjaran, Bandung 40600, Indonesia

Appl. Sci. 2023,13, 7318 4 of 9
5Department of Biotechnology, National Formosa University, Yuanlin 632, Taiwan
∗Correspondence: buntora.pasaribu@unpad.ac.id
Abstract
The significance of intracellular lipid droplets (LDs) was crucial in the mutualis-
tic endosymbiosis between reef-building corals and the dinoflagellate endosymbiont
Symbiodinium
spp. In this study, the researchers aim to investigate and identify LD proteins
present in Symbiodinium residing within the coral host. To achieve this, various methods,
including Discontinuous Percoll gradient centrifugation, were utilized to separate Symbio-
dinium cells from E. glabrescens tentacles. Additionally, the use of Nile red, a fluorescent
probe, demonstrated the accumulation of lipids in the freshly isolated Symbiodinium cells.
We successfully purified stable LDs from the endosymbiotic Symbiodinium cells. The
structural integrity of the Symbiodinium LDs was maintained through mechanisms such
as electronegative repulsion and steric hindrance, potentially facilitated by their surface
proteins. Protein extracts obtained from the purified LDs revealed a prominent protein
band referred to as Symbiodinium lipid droplet protein (SLDP). This study illuminates the
importance of lipid droplets in the endosymbiotic dinoflagellate and their potential role in
the symbiotic relationship between Symbiodinium and reef-building corals.
5. Low-Technology In-Vitro Cultivation of Small Corals Fragments: A Model System
for Clone Production towards Developing Quality R & D
Alex P. Camaya 1, *, Satoko Sekida 2and Kazuo Okuda 2
1
Coastal Resources Management Unit, Bicol University Tabaco Campus, M.H. del Pilar
St., Tayhi, Tabaco City 4511, Albay, Philippines
2Graduate School of Kuroshio Science, Kochi University, 2-5-1 Akebono-cho,
Kochi 780-8520, Japan
∗Correspondence: apcamaya@bicol-u.edu.ph
Abstract
Most of the profound findings in the past investigations of corals have focused on
understanding its aspects of biology vis-a-vis growth-reproduction-dysfunction where
the dynamics of the coexisting host animal and symbiotic algae are crucially challenged
by stress and other factors. In experimental studies where several trials are conducted
in controlled conditions, it is sometimes necessary to establish a knowledge on how to
produce coral cultures as samples to lessen the extraction of healthy colonies from the reef.
In such an event, developing a plausible culture technique holds the key towards capability
in propagating this highly subtle species. In this study, the method of inducing tissue
regeneration in in-vitro systems for scleractinian coral Pocillopora damicornis is essentially
discussed. Following the procedure that was first established in 2014, numbers of isolated
tiny fragments excised from small coral colonies had successfully grown just in glass dishes
with raw seawater at room conditions. These regenerating clones were then utilized as
model samples in observing the growth, cell division, and dysfunction of coral host and
symbiotic cells revealed by light and electron microscopes. With the significant results of the
past experiments derived from utilizing these cultures, this study had demonstrated and
further promote a low-technology system that may produces mass number of viable clones
for various academic and scientific R & Ds, as well as community-based undertakings,
including coral biotechnology, reef restoration, husbandry, MPA conservation, and several
others. However, there are still uncertainties in the success of propagation for other reef-
building species. Hence, this remains a challenging task in future studies.

Appl. Sci. 2023,13, 7318 5 of 9
6. Proteomic Analysis of Acropora cervicornis Exposed to Thermal Acclimation
Samantha Shaw 1,*, Anderson Mayfield 2, Cliff Ross 1and Matthew Gilg 1
1University of North Florida, Jacksonville, FL 32224, USA
2National Museum of Marine Biology and Aquarium, Pingtung 944, Taiwan
∗Correspondence: s.shaw@unf.edu
Abstract
Continuous warming of the oceans has led to increased bleaching events and higher
levels of disease susceptibility. Current scientific efforts have predominately focused on
utilizing transcriptomics to understand thermal stress responses. These efforts have shown
that corals possess plasticity in their response to thermal stress, with certain genotypes
responding well and surviving longer after an initial heat acclimation period. To better
understand the mechanisms by which acclimation occurs, the proteomic response to
thermal acclimation and heat stress of previously studied genotypes was analyzed. The
three least and most plastic genotypes from a prior study were utilized. Fragments of each
genotype were acclimated at 30
◦
C for four days, returned to ambient temperatures (27
◦
C)
for five days, and then heat stressed at 32
◦
C for 2 days. Samples for proteomic analysis
were taken on Day 0, before the experiment began, and then at each temperature transition.
Data will be shown regarding proteomic changes throughout the acclimation process and
subsequent heat stress. Comparisons among genotypes illustrate differences in thermal
acclimation abilities.
7. Heat Tolerance and Symbiodiniaceae Profiles of Acroporids in a Philippine Reef
John Bennedick Quijano *, Jake Ivan P. Baquiran and Cecilia Conaco
Marine Science Institute, University of the Philippines, Diliman, Quezon City, Philippines
∗Correspondence: jbquijano@up.edu.ph
Abstract
The warming of the oceans is decimating coral reefs worldwide. Yet it has been
observed that a proportion of coral populations from the same general reef area, experienc-
ing a similar thermal regime, are able to tolerate heat stress and endure. To understand
the mechanisms underlying coral thermal tolerance, we subjected fragments taken from
30 colonies of Acropora digitifera,A. millepora, and
A. tenuis
to 33
◦
C (high temperature)
versus 29
◦
C (control) for about a week (~2 DHW). Our results revealed inter-individual
differences in heat tolerance in all three coral species, with 70% of A. digitifera, 50% of
A. millepora
, and 30% of A. tenuis colonies showing high tolerance to elevated temperature
(i.e., no bleaching). However, the analysis of associated microalgal symbionts in these
corals using ITS2 sequencing did not demonstrate a clear correlation between differences
in heat tolerance and Symbiodiniaceae composition, suggesting that host-specific factors
should also be examined.
8. The First Cryo-Repository for Coral Larvae: Safeguarding Corals for
Future Generations
Kanokpron Loeslakwiboon 1, Wen-Chung Hsieh 2, Cheng-Liang Huang 3,
Sujune Tsai 4and Chiahsin Lin 1,5,*
1Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan
2He Wei Precision Company Limited, Hsinchu, Taiwan
3Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan
4Department of Post Modern Agriculture, Mingdao University, Chang Hua, Taiwan
5National Museum of Marine Biology & Aquarium, Pingtung, Taiwan
∗Correspondence: chiahsin@nmmba.gov.tw

Appl. Sci. 2023,13, 7318 6 of 9
Abstract
Research on the development of innovative cryobanking techniques will aid coral
restoration and conservation. To date, there are no published studies on long-term cryo-
repository of coral larvae. The aim of this study was to apply our customized freezing
device and cryojig together with vitrification and laser warming techniques to create
the first cryo-repository for coral larvae. In this study, pelagic phase larvae from the
corals Seriatopora caliendrum,Pocillopora verrucosa,P. acuta, and Stylophora pistillata were
used for cryobanking. Three vitrification solutions were formulated with Ficoll and gold
nanoparticles. The results showed that over a thousand coral larvae of S. caliendrum,
P. verrucosa,
P. acuta, and St. pistillata were successfully stored in the cryo-repository. Our
customized innovative technology enabled the long-term cryobanking of coral larvae,
which has never been accomplished before. We believe the methods applied in this study
have the potential to be a critical research and conservation tool for wild reef restoration
and reef habitat diversity.
9. Coral Assessment in Selected Areas in the Verde Island Passage
Enriquo Martin C. Velasquez 1,2,*, Miguel Enrique Ma. A. Azcuna 2,4,
Jonel M. Corral 2,4, Jade Symon A. Binay 1,2 , Karlo L. Pintor 1,2, Rex M. Medina 7,
Katherine Sanchez-Escalona 6, Fhred Kiervin Montante 6and Jayvee Ablaña Saco 1,3,5
1
Verde Island Passage for Oceanographic and Aquatic Life Sciences (VIP CORALS),
Batangas State University, Lobo Campus, Barangay Masaguitsit, Lobo,
Batangas, Philippines
2
Verde Island Passage Center for Oceanographic Research and Aquatic Life Sciences
(VIP CORALS), Batangas State University, Nasugbu Campus, Barangay Bucana,
Nasugbu, Batangas, Philippines
3
College of Arts and Sciences, Batangas State University, Pablo Borbon Campus, Rizal
Ave., Batangas, Philippines
4
College of Arts and Sciences Batangas State University ARASOF, Nasugbu Campus,
Barangay Bucana, Nasugbu, Batangas, Philippines
5
College of Agriculture and Forestry, Batangas State University, Barangay Masaguitsit,
Lobo, Batangas, Philippines
6
Mindoro State University, Main Campus, Alcate, Victoria, Oriental Mindoro, Philippines
7
Scandi Divers Resort, Barangay Sabang, Puerto Galera, Oriental Mindoro, Philippines
∗Correspondence: enriquo.velasquez@g.batstate-u.edu.ph
Abstract
The Verde Island Passage is one of the most biologically diverse ecosystems in the
world. It provides food, livelihood, and other benefits to communities in and around its
waters. However, previous studies conducted here were fragmentary and site-specific.
This study addressed this gap by examining the coral genera and distribution along four
(4) provinces in the Verde Island Passage: Batangas, Marinduque, Occidental Mindoro,
and Oriental Mindoro using the photo transect method. Data were collected during the
northeast monsoon, summer, and southwest monsoon seasons. Each province was assigned
(2) sites classified into highly impacted (HI) and less-impacted (LI) sites. For HI and LI sites,
results showed an average of 21.77% and 27.78% coral cover, 12.70% and 11.98% macroalgae
cover, 24.87% and 30.01% bare rock, 32.06% and 22.89% sand/rubble, respectively. An
average of 25 coral genera were found in Batangas but were mostly dominated by Galaxea
sp. in the highly impacted site and Porites sp. in the less-impacted site. Marinduque
recorded 16 genera, where Porites sp. dominated both highly impacted and less-impacted
sites. Occidental Mindoro recorded 18 genera, where Porites sp. also dominated both
highly impacted and less impacted sites. Lastly, Oriental Mindoro recorded 14 genera
where Galaxea sp. dominated the highly impacted site and Porites sp. dominated the
less-impacted site. Porites sp. Dominance in most of the sites may be linked to their ability

Appl. Sci. 2023,13, 7318 7 of 9
to withstand strong wave action and be more tolerant of stressors and environmental
extremes, such as sedimentation and temperature increase. In general, most of the sites in
all four provinces had good coral cover, including several non-MPA sites. This justifies their
protection, conservation, and possibly elevation of their status to Marine-Protected Areas.
10. Differential Symbiodiniaceae Association with Acropora Humilis Coral While
Rearing in Hatchery Conditions
Suppakarn Jandang 1,5,*, Voranop Viyakarn 1,2, Yuki Yoshioka 3, Chuya Shinzato 3
and Suchana Chavanich 1,2,4
1
Reef Biology Research Group, Department of Marine Science, Faculty of Science,
Chulalongkorn University, Bangkok 10330, Thailand
2Aquatic Resources Research Institute, Chulalongkorn University,
Bangkok 10330, Thailand
3Atmosphere and Ocean Research Institute, The University of Tokyo, Kashiwa,
Chiba 277-8564, Japan
4Center of Excellence for Marine Biotechnology, Department of Marine Science,
Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
5Center for Ocean Plastic Studies, Research Institute for Applied Mechanics,
Kyushu University, Kasuga-Koen, Kasuga, Fukuoka 816-8580, Japan
∗Correspondence: suppakarn.j@riam.kyushu-u.ac.jp
Abstract
Several studies have reported that most coral-Symbiodiniaceae adaptation occurs
when coral faces temperature stress. However, little is known about Symbiodiniaceae pop-
ulation, particularly during coral development in ex-situ conditions. This study provides
the first investigation on the Symbiodiniaceae diversity and community change of reared
Acropora humilis colonies captured in the hatchery system at Samae San Island, Gulf of
Thailand. Coral sexual propagation techniques were conducted to produce experimental
coral colonies. The gamete of A. humilis showed a high fertilization rate of 98.48
±
0.34%.
Nevertheless, the survival rate of reared coral from the early stage to 18 month olds is
declining to 45.96
±
1.31%. Acropora humilis randomly acquired symbionts in early stages
(1 to 3 months old) from an environment associated with three genera (Symbiodinium,
Cladocopium, and Durusdinium) and several Symbiodiniaceae lineages, whereas the com-
munity structure was found stable with almost 100% harbored of Durusdinium D1 at 6 to
18 months old. Reared coral’s offspring was significantly different from Symbiodiniaceae
species compared to wild parent colonies. These results indicated that coral was shown
to change Symbiodiniaceae community composition during development under hatchery
conditions. Knowledge of this study is important to understand the corals-Symbiodiniaceae
association, the status of coral reef prediction, and coral conservation in Thailand.
11. Coral Restoration Program in the Bicol Region, the Philippines:
Experiences and Challenges
Antonino B. Mendoza 1,*, Satoshi Kubota 2and Plutomeo M. Nieves 1
1Bicol University Tabaco Campus, M.H. del Pilar St., Tayhi, Tabaco City 4511,
Albay, Philippines
2Department of Agriculture, Kochi University, Nankoku, Kochi 783-8502, Japan
∗Correspondence: ajbmendoza@bicol-u.edu.ph
Abstract
Several coral restoration programs in Bicol Region, Philippines were conducted by
various sectors in the hope to help improve the ailing coral reefs in the area. In this pa-
per, we discussed the experiences and challenges in the implementation of the program.
The program did not only intend to rehabilitate degraded reef areas but also to showcase

Appl. Sci. 2023,13, 7318 8 of 9
to the communities its viability as a community-based endeavor, allowing partnership
with different sectors of the society. Various rehabilitation methods were discussed from
low-cost to high-cost designs and the possibilities of other economic opportunities. Chal-
lenges enumerated from natural calamities to social issues were also pointed out. Overall,
coral rehabilitation program in the region showed high acceptability from the community.
However, sustainability has been observed as the problem.
12. Spatiotemporal Lipid Profiling during Coral Embryogenesis
Federica Buttari 1,2, Sujune Tsai 3, Zhi-Hong Wen 1and Chiahsin Lin 2,4, *
1
Department of Marine Biotechnology and Resources, National Sun Yat-Sen University,
Kaohsiung, Taiwan
2National Museum of Marine Biology & Aquarium, Pingtung, Taiwan
3Department of Post-Modern Agriculture, Mingdao University, Chang Hua, Taiwan
4Institute of Marine Biology, National Dong Hwa University, Pingtung, Taiwan
∗Correspondence: chiahsin@nmmba.gov.tw
Abstract
To evaluate the variation in the content and composition of lipids and understand
their role during the embryonic developmental stages, major lipid classes: sterol ester
(SE), wax ester (WE), triacylglycerol (TAG), cholesterol, phosphatidylethanolamine (PE),
phosphatidylcholine (PC), lysophosphatidylcholine (LPC), and fatty acids were assessed
on the octopus coral Galaxea fascicularis in this study. The gametes and embryos were
homogenized with a mixture of chloroform and methanol, obtaining a biphasic system in
which the chloroform layer contained the lipids. Thin layer chromatography (TLC), transes-
terification to fatty acid methyl ester (FAME), and gas chromatography mass spectrometry
were used to determine the lipid and fatty acids content. The results showed that SE,
WE, and TAG, altered in content during the different developmental stages, although not
constantly, while cholesterol, PC, PE, and LPC remained unchanged in content throughout
the course of the various developmental stages. Cholesterol and PC were the dominant
lipids in all stages, followed by WE and LPC, which has medium lipid contents, whereas
the contents of TAG, PE, and SE were significantly lower than those of the aforementioned
lipid classes. In contrast with the lipid profile of the oocyte, sperm exhibited a much lower
content of WE and the non-appearance of SE. However, the rest of the lipid classes showed
no statistical difference in content. As concerned the content of fatty acids, there was a
peak of concentration in the early stages of embryonic development with the prevalence
of saturated fatty acids (SFA). On the other hand, there was a low content of fatty acids
at the sperm level. This study revealed that lipid content and composition may fluctuate
between embryonic developmental stages. The findings will undoubtedly assist future
lipid research in other coral species.
13. Benthic Community Structures with Varying Sedimentation and Water Quality
Conditions in Mabini, Batangas
Danilo A. Leyble Jr. *, Riza Maree D. C. Rapada, Ethel C. Wagas
and Milette U. Mendoza-Pascual
Department of Environmental Science, Ateneo de Manila University, Quezon City, Philippines
∗Correspondence: danilo.leyble@obf.ateneo.edu
Abstract
Marine-protected areas have long been used as tools for protecting reef ecosystems
in the Philippines. No-take marine zones are established to limit direct human activity,
which is one of the primary causes for the degradation of coral reefs and reef-associated
organisms. However, Philippine reefs are not only at risk from direct human activities in
reef areas but from nonpoint sources such as sedimentation and poor water quality. This

Appl. Sci. 2023,13, 7318 9 of 9
study assessed and compared the hard coral cover with physical and chemical water quality
parameters, such as sedimentation rate, TSS, pH, nitrate concentration, and phosphate
concentrations or reefs inside and outside a marine-protected area. Samples were obtained
through the photo-transect method, use of sediment traps, and standard methods for
water quality assessment. Hard corals were significantly higher in the MPA, although no
spatial differences were observed for sedimentation rate and water quality. Sedimentation
rates, TSS, and nutrient concentrations were significantly higher during the wet season,
while no temporal differences were observed for the benthic cover. Principal Component
Analysis (PCA) revealed that sites were differentiated through hard coral cover, pH, and
TSS concentrations, while seasons are influenced by sedimentation rates and nutrient
concentrations. This study highlights the need for a ridge-to-reef approach in conserving
coastal ecosystems. The current management practices of the MPA do not entirely inhibit
the effects of sedimentation and nutrient pollution in the reef, which occur when run-offs
from land-based activities such as infrastructure developments happen along the coast
without proper management measures being set in place.
14. Characteristics of Black Pigment Released from the Octocoral Sinularia flexibilis
Lian-Di Lin 1, Yu-Chia Chang 2and Hsing-Hui Li 1,3,*
1National Museum of Marine Biology and Aquarium, Pingtung 944013, Taiwan
2
Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry
Technology, College of Human Ecology, Chang Gung University of Science and
Technology, Taoyuan 333324, Taiwan
3
Department of Marine Biotechnology and Resources, National Sun Yat-Sen University,
Kaohsiung 804201, Taiwan
∗Correspondence: hhli@nmmba.gov.tw
Abstract
In our previous study, we determined that the octocoral Sinularia flexibilis released a
black pigment that stained the operator’s hands black when it was cut during the process
of asexual propagation. This black pigment was water-insoluble, sticky, and constantly
entangled with Symbiodiniaceae. At the time, we were unable to identify any solvents
that could dissolve the pigment. In the present study, we established a method to purify
the pigment and remove the contamination of Symbiodiniaceae to better understand the
pigment’s nature. We determined that the pigment can be dissolved in a strong basic
solution, 1M sodium peroxide (pH value 14). We confirmed that the black pigment is
not melanin by using Fourier–Transform Infrared Spectroscopy and the Fontana–Masson
melanin stain. By staining the tissue sections, we determined that the black pigment is
generated in the endoderm of the coral, then moved to the ectoderm, and finally released
outside of the corals. The structure and function of this black pigment merit further study
for its potential to become a new black dye for human industries.
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