coral growth and monitoring using the micro-device system

Un sistema combinato di micro-dispositivi per promuovere e monitorare la crescita dei coralli

The deterioration of coral reef ecosystems has been occurring worldwide over the past 70 years. Considering all the major coral areas across Central America1, Southeast Asia2,3,4,5,6, Australia7,8, and East Africa9, the global coverage of coral reefs has halved since the 1950s10. This mass loss of coral reefs has resulted in ecological and economic problems. For example, by tracing the presence/absence and abundance of all kinds of coral-dependent fishes for 8 years, researchers concluded that the coral decline has directly caused a substantial decrease in fish biodiversity and abundance in Papua New Guinea11. This result proved that the coral decline can not only undermine coral reef-based biological systems but also reduce fishery incomes.
Over decades of field surveys, including direct monitoring, remote sensing, and data comparison, the scientific community has identified several factors causing the mass coral decline. One major reason for the mass coral decline is coral bleaching caused by high seawater temperatures12,13. By combining bleaching and meteorological records, scientists have concluded that coral bleaching is happening more frequently in El Ni&#241;o-Southern Oscillation phases14. Another reason for the coral decline is ocean acidification. Owing to the increased CO2 concentration in both the atmosphere and seawater, calcium carbonate dissolves faster than before, causing downscale net coral reef calcification15. Indeed, it has been concluded that when the CO2 concentration in the atmosphere reaches above 500 ppm, tens of millions of people will suffer, and the coral reefs will be at risk of significant deterioration and symbiodinium detachment16,17. There are other factors that can also affect coral survival, such as inshore pollutants causing or accelerating coral decline. Researchers in Hawaii measured the carbon, oxygen, and nitrogen isotopes in corals, along with the dissolved inorganic carbonate and the related nutrients (NH4+, PO43-, NO2&#8722;, and NO3&#8722;), and concluded that pollution from the land magnified the coastal acidification and bioerosion of corals18. Further to pollution, urbanization also endangers coral survival and causes relatively low architectural complexity in corals, as revealed by a study on the coral survival status in Singapore, Jakarta, Hong Kong, and Okinawa. Thus, the impact of anthropogenic stressors and the superimposed effects of climate change are leading to widespread reduced biodiversity on coral reefs and an associated decline in coral ecological function and resilience19.
It should also be noted that a large number of microorganisms participate in the physiological functions of corals, including nitrogen fixation, chitin decomposition, the synthesis of organic compounds, and immunity20, and these microorganisms should, thus, be included when considering coral reef deterioration. In natural environments, such as coral reefs, many factors cause hypoxic or anoxic conditions, including insufficient water circulation, algal exudate, and algal overgrowth. This phenomenon negatively affects the population distributions of coral and coral-related microorganisms. For example, Vietnamese scientists found that in Nha Trang, Phu Quoc, and Ujung Gelam, bacterial composition in the coral Acropora Formosa could be affected by dissolved oxygen at different locations21. Researchers in the United States explored hypoxic or anoxia conditions in corals and found that algal exudates can mediate microbial activity, leading to localized hypoxic conditions, which may cause coral mortality in the direct vicinity. They also found that corals could tolerate reduced oxygen concentrations but only above a given threshold determined by a combination of the exposure time and oxygen concentration22. Researchers in India found that when Noctiluca scintillans algae bloomed, the dissolved oxygen decreased to 2 mg/L. Below this concentration, about 70% of Acropora montiporacan died because of hypoxic conditions23.
All the abovementioned facts and factors suggest that environmental change leads to the deterioration of coral reefs. To culture and study reef corals under certain conditions, it is important to accurately and comprehensively build up a controllable microscopic environment for reef corals to inhabit. Normally, scientists focus on temperature, light, water flow, and nutrients. However, other features, such as the dissolved oxygen concentration, microorganism abundance, and microorganism diversity in the seawater, are commonly ignored. To this end, our group has explored the possibility of applying small equipment to culture coral polyps in a relatively controlled environment24,25. In this work, we designed and built up a modular micro-device system for coral culture. This modular micro-device system can provide a controllable micro-environment in terms of the temperature, light spectrum, dissolved oxygen concentration, nutrients, and microorganisms, etc., and has the capacity for expansion and upgrade.
Modules and functions of the device 
The micro-device system was inspired by the Berlin system26, but no live rocks are used in the current system. As shown in Figure 1, the current system comprises six main modules, two brushless motor pumps, one gas pump, one flow-through UV lamp, one power supply, certain electronic control components, and the related wires and screws. The six main modules include a seawater store module (with an air pump and temperature sensor), a temperature control module, an algae purification module, a microbial purification module, an activated charcoal purification module, and a coral culture module.
Device architecture 
As shown in Figure 2 and Figure 3, the overall micro-device system can horizontally be divided into two compartments with a temperature control module in between. For safety reasons, all the seawater-containing modules and parts are placed in the left compartment, named the culture compartment. The other electronic parts are placed in the right compartment, named the electronic compartment. Both compartments are sealed or packaged within shells. The temperature control module is fixed in a divider plate in between. The shell of the culture compartment includes a baseboard and three screw-fixing plates. This design ensures compartment tightness and facilitates the operation of the system. Additionally, the tightness favors accurate temperature control. The shell of the electronic compartment includes a baseboard, two screw-fixing plates, and one front control panel.
Water circulation 
An inner and outer seawater circulation loop connected to the seawater store module was pre-designed. The inner circulation loop successfully connects the seawater store module, temperature control module, flow-through UV lamp, algae purification module, and microbial purification module. This circulation loop aims to provide suitable physiochemical and physiological seawater conditions for the corals, and no frequent maintenance is needed. The algae purification module contains Chaetomorpha algae, which absorbs the extra nutrients (nitrate and phosphate) in the water. The microbial purification module contains the bacterial culture substrate, which cultivates the microbiome to transfer nitrite and ammonium into nitrate for water purification. All these modules need to be replaced only under critical circumstances.
The outer circulation loop successively connects the seawater store module, coral culture module, and activated charcoal module. This circulation loop aims to provide light, tightness, water current, and high seawater quality to the corals. The seawater can be refreshed through a water inlet and a water outlet. Additives are added through a three-way valve, and the seawater sample can also be extracted from this valve for inspection. Air can be pumped in through an air inlet and discharged from an air outlet.
Electronic design 
A 220 V AC power supply with a switch and a fuse is used for the whole system. The input power is divided into four branches. The first branch goes to a 12 V DC power supply, which directly powers the heating panel, cooling panel, and cooling fan. This branch also indirectly powers two pumps and two lighting panels through a four-channel DC transformer. The second branch goes to a PID temperature controller. The third branch goes to an air pump power supply. The last branch connects to a UV lamp power supply. A solid-state relay connects the PID temperature controller and the cooling panel in the temperature control module. A regular relay is used to connect the PID temperature controller and the heating panel. The four-channel DC transformer converts the voltage to that required.
There are two control panels on the right part of the system. There are four switches and one controller for the UV lamp on the top panel, including a main power switch, a UV lamp power switch, an air pump switch, and a temperature control switch. The main power switch controls the 12 V power supply of the system.
A PID temperature controller, a cycle timer, a four-channel DC transformer, and a three-channel timer are on the front panel. The PID temperature controller adjusts the water temperature by controlling the heating and cooling panels in the temperature control module. The temperature control module only works when the inner circulation pump is working and the water is flowing past the temperature control module. The cycle timer is connected to the air pump power line. Its purpose is to assign the working time period to the air pump. There is a three-channel timer deployed in the electronic compartment too. This timer controls the work time period for the air pump, coral light, and algae light.

Autore in primo piano: Avanzamento della coltura dei coralli - Creazione di un sistema di microambiente semi-quantitativamente controllato per contrastare le attuali limitazioni 

Un sistema integrato di micro-dispositivi per la crescita e il monitoraggio dei coralli

Watch this Scientific Journal Video about Coral Growth and Monitoring Using the Micro-Device System at JoVE.com

Coral Growth and Monitoring Using the Micro-Device System  

Crescita e monitoraggio dei coralli con il sistema di micro-dispositivi

Dopo aver impostato il sistema di microazionamento, aggiungere un millilitro della soluzione di origine del microbioma commerciale in 500 millilitri di acqua di mare mescolando. Successivamente, iniettare 50 millilitri della soluzione diluita e 10 microlitri della soluzione nutrizionale commerciale per coralli nel sistema di circolazione. Accendere la pompa di circolazione interna e accendere la pompa dell'aria per coltivare il microbioma per 21 giorni.Taglia i rami di corallo di roccia in una lunghezza da tre a cinque centimetri. Incolla questi rami di corallo su basi di supporto di corallo stampate in 3D. Rimetti i campioni di rami di corallo nel serbatoio originale dell'acqua di mare per un minimo di sette giorni per il recupero.Fissare la base di supporto in corallo sull'unità di rotazione utilizzando la colla. Assemblare il modulo di coltura del corallo e collegarlo al circuito di circolazione esterno. Posiziona la fotocamera sulla parte superiore dello studio e cattura le immagini da una vista verticale.Successivamente, posiziona il modulo di coltura del corallo nel mini studio fotografico con il corallo posizionato al centro e in basso. Cattura immagini di corallo ruotando la maniglia esterna. Il modulo di coltura dei coralli ha controllato efficacemente la temperatura dell'acqua di mare e ha impedito le fluttuazioni di temperatura, che possono influenzare fortemente la sopravvivenza dei coralli.I tentacoli dei coralli campione si estendevano per oltre un mese, indicando che il sistema forniva un ambiente di sopravvivenza adatto per il corallo.

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Research

JoVE Journal

Bioengineering

Coral Growth and Monitoring Using the Micro-Device System

111 Views. Dopo aver impostato il sistema di microazionamento, aggiungere un millilitro della soluzione di origine del microbioma commerciale in 500 millilitri di acqua di mare mescolando. Successivamente, iniettare 50 millilitri della soluzione diluita e 10 microlitri della soluzione nutrizionale commerciale per coralli nel sistema di circolazione. Accendere la pompa di circolazione interna e accendere la pompa dell'aria per coltivare il microbioma per 21 giorni.Taglia i rami di corallo di rocci...

Video: Coral Growth and Monitoring Using the Micro-Device System  

An Integrated Micro-Device System for Coral Growth and Monitoring

Corals are fundamental organisms in marine and coastal ecosystems. With the advancement of coral protection research in recent years, precise control of the coral culture environment is highly in demand for coral conservation and study. Here, we developed a semi-closed coral culture micro-device system as a multi-functional platform, which can provide accurate and programmable temperature control, a sterile initial environment, long-term stable water quality, an adjustable dissolved oxygen concentration, and a customized light spectrum for corals. Owing to the modular design, the coral culture system can be upgraded or modified by installing desirable new modules or removing existing ones. Currently, under appropriate conditions and with proper system maintenance, the sample corals can survive for at least 30 days in a healthy state. Furthermore, due to the controllable and sterile initial environment, this coral culture system can support research into the symbiotic relationship between corals and associated microorganisms. Therefore, this micro-device system can be applied to monitor and investigate sea corals in a relatively quantitative manner.

This protocol describes the development of a modular controllable micro-device system that can be applied for the long-term culturing and monitoring of sea corals.

Corals are fundamental organisms in marine and coastal ecosystems. With the advancement of coral protection research in recent years, precise control of ...