Name: Scaling Microalgal Biotechnology for Enhanced Biomethane Production (Video) | JoVE
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We thank DGAPA UNAM project number IT100423 for the partial funding. We also thank PROAN and GSI for allowing us to share technical experiences about their photosynthetic biogas upgrading full installations. The technical support of Pedro Pastor Hern&#225;ndez Guerrero, Carlos Martin Sigala, Juan Francisco D&#237;az M&#225;rquez, Margarita Elizabeth Cisneros Ortiz, Roberto Sotero Briones M&#233;ndez and Daniel de los Cobos Vasconcelos is highly appreciated. A part of this research was done at IIUNAM Environmental Engineering Laboratory with an ISO 9001:2015 certificate.

1. System set-up
NOTE: A piping and instrumentation diagram (P&#38;ID) of the system described in this protocol is shown in Figure 2.
<ol>
	<li>Reactor set-up
	<ol>
		<li>Prepare the ground by leveling and compacting it to improve reactor stability.</li>
		<li>On an open field, dig two elongated holes and 3 m from the end, further dig a 3 m2 and 1 m deep hole (known as an aeration well).</li>
		<li>Place two HRAPs (<strong class="xfig...

Microalgal Cultivation for Biomethane Obtention from Swine Waste Biogas

<ol>
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Manuel de culture artisanale de spiruline Available from: <a target="_blank" href="https://www.scribd.com/document/513003475/Manuel-de-Culture-Artisanale-de-Spiruline">https://www.scribd.com/document/513003475/Manuel-de-Culture-Artisanale-de-Spiruline</a> (2006)</li><li>Lu, L., Yang, G., Zhu, B., Pan, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+three+quantitating+methods+of+microalgal+biomass.">A comparative study on three quantitating methods of microalgal biomass.</a> Indian J Geo-Mar Sci. 46, 2265-2272 (2017).</li><li>Sukarni, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermogravimetric+analysis+of+the+combustion+of+marine+microalgae+Spirulina+platensis+and+its+blend+with+synthetic+waste.">Thermogravimetric analysis of the combustion of marine microalgae Spirulina platensis and its blend with synthetic waste.</a> Heliyon. 6 (9), e04902 (2020).</li><li>Kundu, S., Zanganeh, J., Moghtaderi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+on+understanding+explosions+from+methane-air+mixture.">A review on understanding explosions from methane-air mixture.</a> J Loss Prev Process Ind. 40, 507-523 (2016).</li><li>Serejo, M. 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A review.</a> Renew Sustain Energy Rev. 137, 137 (2021).</li><li>lvarez-Gonz&#225;lez, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+microalgae+grown+in+wastewater+reduce+the+use+of+inorganic+fertilizers.">Can microalgae grown in wastewater reduce the use of inorganic fertilizers.</a> J Environ Manage. 323, 116224 (2022).</li><li>Deepika, P., MubarakAli, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+and+assessment+of+microalgal+liquid+fertilizer+for+the+enhanced+growth+of+four+crop+plants.">Production and assessment of microalgal liquid fertilizer for the enhanced growth of four crop plants.</a> Biocatal Agric Biotechnol. 28, 101701 (2020).</li><li>. Perspectives for a european standard on biomethane: a Biogasmax proposal Available from: <a target="_blank" href="https://trimis.ec.europa.eu/sites/default/files/project/documents/20120601_135059_69928_d3_8_new_lmcu_bgx_eu_standard_14dec10_vf__077238500_0948_26012011.pdf">https://trimis.ec.europa.eu/sites/default/files/project/documents/20120601_135059_69928_d3_8_new_lmcu_bgx_eu_standard_14dec10_vf__077238500_0948_26012011.pdf</a> (2010)</li><li>. Biomethane - Oxygen Content Assessment Available from: <a target="_blank" href="https://www.gasnetworks.ie/docs/corporate/gas-regulation/Oxygen-concentration-report-17985-AI-RPT-001-Rev-5-Biomethane-review-Penspen.pdf">https://www.gasnetworks.ie/docs/corporate/gas-regulation/Oxygen-concentration-report-17985-AI-RPT-001-Rev-5-Biomethane-review-Penspen.pdf</a> (2018)</li><li>. European biomethane standards for grid injection and vehicle fuel use Available from: <a target="_blank" href="https://www.biosurf.eu/wordpress/wp-content/uploads/2015/06/9.-Arthur_Wellinger.pdf">https://www.biosurf.eu/wordpress/wp-content/uploads/2015/06/9.-Arthur_Wellinger.pdf</a> (2017)</li><li>. NORMA Oficial Mexicana NOM-001-SECRE-2010, Especificaciones del gas natural (cancela y sustituye a la NOM-001-SECRE-2003, Calidad del gas natural y la NOM-EM-002-SECRE-2009, Calidad del gas natural durante el periodo de emergencia severa) Available from: <a target="_blank" href="https://www.dof.gob.mx/normasOficiales/3997/sener/sener.html">https://www.dof.gob.mx/normasOficiales/3997/sener/sener.html</a> (2010)</li><li>Sharifian, R., Wagterveld, R. M., Digdaya, I. A., Xiang, C., Vermaas, D. 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Throughout the years, this algal technology has been tested and used as an alternative to the harsh and expensive physicochemical techniques to purify biogas. Particularly, the Arthrospira genus is widely used for this specific purpose, along with Chlorella. There are few methodologies, however, that are made on a semi-industrial scale, which adds value to this procedure.
It is critical to maintain lower O2 concentrations by using the proper L/G ratio; however, thi...

In recent years, several technologies have emerged to purify biogas to biomethane, promoting its use as non-fossil fuel, therefore mitigating undesairable methane emissions1. Air pollution is a problem that affects most of the world&#39;s population, particularly in urbanized areas; ultimately, around 92% of the world&#39;s population breathe polluted air2. In Latin America, the air pollution rates are mostly created by the use of fuels, whereby in 2014, 48% of the air pollution was brought on by the electricity and heat production sector3.
In the last decade, more and more studies on the relationship between pollutants in the air and the increase in mortality rates have been proposed, arguing that there is a strong correlation between both datasets, particularly in children populations.
As a way to avoid the continuation of air pollution, several strategies have been proposed; one of these is the use of renewable energy sources, including wind turbines and photovoltaic cells, which diminish the CO2 release into the atmosphere4,5. Another renewable energy source comes from biogas, a byproduct of the anaerobic digestion of organic matter, produced along with a liquid organic digestate6. This gas is composed of a mixture of gasses, and their proportions depend on the source of organic matter used for anaerobic digestion (sewage sludge, cattle manure, or agro-industrial biowaste). Generally, these proportions are CH4 (53%-70%vol), CO2 (30%-47%vol), N2 (0%-3%vol), H2O (5%-10%vol), O2 (0%-1%vol), H2S (0-10,000 ppmv), NH3 (0-100 ppmv), hydrocarbons (0-200 mg/m3) and siloxanes (0-41 mg/m3)7,8,9, where the scientific community is interested in the methane gas since this is the renewable energetic component of the mixture.
However, biogas cannot be simply burned as obtained because the byproducts of the reaction can be harmful and contaminant; this raises the need to treat and purify the mixture to increase the percentage of methane and decrease the rest, essentially converting it into biomethane10. This process is also known as upgrading. Even though, currently, there are commercial technologies for this treatment, these technologies have several economic and environmental drawbacks11,12,13. For example, systems with activated carbon and water washing (ACF-WS), pressure water washing (PWS), gas permeation (GPHR), and pressure swing adsorption (PSA) present some economic or other drawbacks of environmental impact. A viable alternative (Figure 1) is the use of biological systems such as those that combine microalgae and bacteria grown in photobioreactors; some advantages include the simplicity of design and operation, the low operating costs, and its environmentally friendly operations and byproducts10,13,14. When biogas is purified to biomethane, the latter can be used as a substitute for natural gas, and the digestate can be implemented as a source of nutrients to support microalgae growth in the system10.
A method widely used in this upgrading procedure is the growth of microalgae in open raceway photoreactors coupled with an absorption column due to the lower operation costs and the minimal investment capital needed6. The most used type of raceway reactor for this application is the high-rate algal pond (HRAP), which is a shallow raceway pond where the circulation of the algal broth occurs via a low-power paddle wheel14. These reactors need large areas for their installation and are very susceptible to contamination if used in outdoor conditions; in biogas purification processes, it is advised to use alkaline conditions (pH &#62; 9.5) and the use of algal species that thrive in higher pH levels to enhance the removal of CO2 and H2S while avoiding contamination15,16.
This research aimed to determine the biogas treatment efficiencies and final production of biomethane using HRAP photobioreactors coupled with an absorption-desorption column system and a microalgae consortium.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1&#34; rotameter</td>
			<td>CICLOTEC</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#34; rotameter</td>
			<td>GPI</td>
			<td>A10-LMA100IA1</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorption tank</td>
			<td>EFISA</td>
			<td>Made under previous design</td>
			<td></td>
		</tr>
		<tr>
			<td>Air blower (2.35 HP)</td>
			<td>Elmo Rietschle</td>
			<td>2BH11007AH01</td>
			<td></td>
		</tr>
		<tr>
			<td>Biogas blower (2 HP)</td>
			<td>Elmo Rietschle</td>
			<td>2BH11007AH01</td>
			<td></td>
		</tr>
		<tr>
			<td>Biogas composition measure</td>
			<td>Geotech</td>
			<td>BIOGAS 5000</td>
			<td></td>
		</tr>
		<tr>
			<td>Data-acquisition device</td>
			<td>LabJack Co.</td>
			<td>U3-LV</td>
			<td></td>
		</tr>
		<tr>
			<td>Diffuser tubes</td>
			<td>Aero-Tube</td>
			<td>C3060AR</td>
			<td></td>
		</tr>
		<tr>
			<td>DO sensor</td>
			<td>Applisens</td>
			<td>Z10023525</td>
			<td></td>
		</tr>
		<tr>
			<td>Dodecahydrated trisodium phosphate&#160;</td>
			<td>Quimica PIMA</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Dodecahydrated trisodium phosphate&#160;</td>
			<td>Fermont</td>
			<td>35963</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Durapore membrane (45 &#181;m)</td>
			<td>MerckMillipore</td>
			<td>HVLP04700&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric motor 1.5 HP</td>
			<td>Weg</td>
			<td>00158ET3ERS56C</td>
			<td></td>
		</tr>
		<tr>
			<td>Ferrous sulfate heptahydrate</td>
			<td>Agroquimica Samet</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Ferrous sulfate heptahydrate</td>
			<td>Fermont</td>
			<td>63593</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Geomembrane</td>
			<td>GEOSINCERE</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium sulfate heptahydrate</td>
			<td>Tepeyac</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Magnesium sulfate heptahydrate</td>
			<td>Fermont</td>
			<td>63623</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Paddle wheel</td>
			<td>GSI</td>
			<td>Made under previous design</td>
			<td></td>
		</tr>
		<tr>
			<td>pH sensor</td>
			<td>Van London pHoenix</td>
			<td>715-772-0041</td>
			<td></td>
		</tr>
		<tr>
			<td>Portable screen</td>
			<td>Rasspberry</td>
			<td>Pi 3 B+</td>
			<td></td>
		</tr>
		<tr>
			<td>Recirculation centrifugal pump (1.5 HP)</td>
			<td>Aquapak&#160;</td>
			<td>ALY 15</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Industria del alcali</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Fermont</td>
			<td>12903</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>Sal Colima</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>Fermont</td>
			<td>24912</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Sodium nitrate</td>
			<td>Vitraquim</td>
			<td>N/A</td>
			<td>Fertilizer grade (greenhouse and experior use)</td>
		</tr>
		<tr>
			<td>Sodium nitrate</td>
			<td>Fermont</td>
			<td>41903</td>
			<td>Analytical grade (Used in cultures inside the laboratory)</td>
		</tr>
		<tr>
			<td>Storing program (pH, DO)</td>
			<td>&#160;Python Software Foundation&#160;</td>
			<td>Python IDLE 2.7</td>
			<td></td>
		</tr>
		<tr>
			<td>Tedlar bags</td>
			<td>SKC Inc.</td>
			<td>232-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature recorder</td>
			<td>T&#38;D</td>
			<td>TR-52i</td>
			<td></td>
		</tr>
		<tr>
			<td>UV-Vis Spectrophotometer</td>
			<td>ThermoFisher Scientific instrument</td>
			<td>GENESYS 10S&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>EVAR</td>
			<td>EV-40</td>
			<td></td>
		</tr>
	</tbody>
</table>

biogas purification through the use of a microalgae-bacterial system in semi-industrial high rate algal ponds

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of polluting gases such as carbon dioxide and hydrogen sulfide to increase the content of methane. In this study, we used a microalgal cultivation technology to treat and purify biogas produced from organic waste from the swine industry to obtain ready-to-use biomethane. For cultivation and purification, two 22.2 m3 open-pond photobioreactors coupled with an absorption-desorption column system were set up in San Juan de los Lagos, Mexico. Several recirculation liquid/biogas ratios (L/G) were tested to obtain the highest removal efficiencies; other parameters, such as pH, dissolved oxygen (DO), temperature, and biomass growth, were measured. The most efficient L/Gs were 1.6 and 2.5, resulting in a treated biogas effluent with a composition of 6.8%vol and 6.6%vol in CO2, respectively, and removal efficiencies for H2S up to 98.9%, as well as maintaining O2 contamination values of less than 2%vol. We found that pH greatly determines CO2 removal, more so than L/G, during cultivation because of its participation in the photosynthetic process of microalgae and its ability to vary pH when solubilized due to its acidic nature. DO, and temperature oscillated as expected from the light-dark natural cycles of photosynthesis and the time of day, respectively. Biomass growth varied with CO2 and nutrient feeding as well as reactor harvesting; however, the trend remained primed for growth.

Biogas Purification through the use of a Microalgae-Bacterial System in Semi-Industrial High Rate Algal Ponds

Following the protocol, the system was built, tested, and inoculated. The conditions were measured and stored, and the samples were taken and analyzed. The protocol was performed a year, starting in October 2019 and lasting until October 2020. It is important to mention that from here onwards, the HRAPs will be referred to as RT3 and RT4.
Biomethane productivity 
In order to determine the conditions that promote the highest H2S and CO2 removal and, c...

Watch this Scientific Journal Video about Scaling Microalgal Biotechnology for Enhanced Biomethane Production at JoVE.com

Scaling Microalgal Biotechnology for Enhanced Biomethane Production (Video) | JoVE

Air pollution impacts the quality of life of all organisms. Here, we describe the use of microalgae biotechnology for the treatment of biogas (simultaneous removal of carbon dioxide and hydrogen sulfide) and the production of biomethane through semi-industrial open high-rate algal ponds and subsequent analysis of treatment efficiency, pH, dissolved oxygen, and microalgae growth.

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of ...