Name: methane hydrate crystallization on sessile water droplets
Uploaded: 2021-05-26T15:00:00
Description: Watch this Scientific Journal Video about Methane Hydrate Crystallization on Sessile Water Droplets at JoVE.com

A nasa Exobiology Grant 80NSSC19K0477 financiou esta pesquisa. Agradecemos a William Waite e Nicolas Espinoza por discussões valiosas.

1. Projete, valide e máquina a célula de pressão.
<ol><li>Projete a célula para permitir a visualização direta da formação de hidrato a partir de uma gotícula de água. Certifique-se de que a célula tenha uma câmara principal com janela de safira e quatro portas para entrada de fluido/gás, saída, luz e fios(Figura 1). Crie o design final em software de design de engenharia(Figura Suplementar S1).</li>
	<li>Para verificar se a célula de pressão é segura sob ...

<ol>
	<li>Bohrmann, G., Torres, M. E., Schulz, H. D., Zabel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gas+hydrates+in+marine+sediments.">Gas hydrates in marine sediments.</a> Marine Geochemistry. , 481-512 (2006).</li><li>Ruppel, C. D., Kessler, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+interaction+of+climate+change+and+methane+hydrates.">The interaction of climate change and methane hydrates.</a> Reviews of Geophysics. 55 (1), 126-168 (2017).</li><li>Hammerschmidt, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+gas+hydrates+in+natural+gas+transmission+lines.">Formation of gas hydrates in natural gas transmission lines.</a> Industrial and Engineering Chemistry. 26, 851-855 (1934).</li><li>Ke, W., Kelland, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetic+hydrate+inhibitor+studies+for+gas+hydrate+systems:+a+review+of+experimental+equipment+and+test+methods.">Kinetic hydrate inhibitor studies for gas hydrate systems: a review of experimental equipment and test methods.</a> Energy &amp; Fuels. 30 (12), 10015-10028 (2016).</li><li>Kelland, M. A. <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+of+kinetic+hydrate+inhibitors+from+an+environmental+perspective.">A review of kinetic hydrate inhibitors from an environmental perspective.</a> Energy &amp; Fuels. 32 (12), 12001-12012 (2018).</li><li>Walker, V. K., 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=Antifreeze+proteins+as+gas+hydrate+inhibitors.">Antifreeze proteins as gas hydrate inhibitors.</a> Canadian Journal of Chemistry. 93 (8), 839-849 (2015).</li><li>Bruusgaard, H., Lessard, L. D., Servio, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphology+study+of+structure+I+methane+hydrate+formation+and+decomposition+of+water+droplets+in+the+presence+of+biological+and+polymeric+kinetic+inhibitors.">Morphology study of structure I methane hydrate formation and decomposition of water droplets in the presence of biological and polymeric kinetic inhibitors.</a> Crystal Growth &amp; Design. 9 (7), 3014-3023 (2009).</li><li>Jung, J. W., Espinoza, D. N., Santamarina, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+and+phenomena+relevant+to+CH4-CO2+replacement+in+hydrate-bearing+sediments.">Properties and phenomena relevant to CH4-CO2 replacement in hydrate-bearing sediments.</a> Journal of Geophysical Research. 115 (10102), 1-16 (2010).</li><li>Chen, X., Espinoza, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ostwald+ripening+changes+the+pore+habit+and+spatial+variability+of+clathrate+hydrate.">Ostwald ripening changes the pore habit and spatial variability of clathrate hydrate.</a> Fuel. 214, 614-622 (2018).</li><li>DuQuesnay, J. R., Diaz Posada, M. C., Beltran, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+gas+hydrate+reactor+design:+3-in-1+assessment+of+phase+equilibria,+morphology+and+kinetics.">Novel gas hydrate reactor design: 3-in-1 assessment of phase equilibria, morphology and kinetics.</a> Fluid Phase Equilibria. 413, 148-157 (2016).</li><li>Udegbunam, L. U., DuQuesnay, J. R., Osorio, L., Walker, V. K., Beltran, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+equilibria,+kinetics+and+morphology+of+methane+hydrate+inhibited+by+antifreeze+proteins:+application+of+a+novel+3-in-1+method.">Phase equilibria, kinetics and morphology of methane hydrate inhibited by antifreeze proteins: application of a novel 3-in-1 method.</a> The Journal of Chemical Thermodynamics. 117, 155-163 (2018).</li><li>Espinoza, D. N., Santamarina, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water-CO2-mineral+systems:+Interfacial+tension,+contact+angle,+and+diffusion+-+Implications+to+CO2+geological+storage.">Water-CO2-mineral systems: Interfacial tension, contact angle, and diffusion - Implications to CO2 geological storage.</a> Water Resources Research. 46 (7537), 1-10 (2010).</li><li>Sloan, E. D., Koh, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clathrate Hydrates of Natural Gases. 3rd edn. , (2007).</li><li>Makogon, I. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Hydrates of natural gas. , 125 (1981).</li><li>Johnson, A. M., 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=Mainly+on+the+plane:+deep+subsurface+bacterial+proteins+bind+and+alter+clathrate+structure.">Mainly on the plane: deep subsurface bacterial proteins bind and alter clathrate structure.</a> Crystal Growth &amp; Design. 20 (10), 6290-6295 (2020).</li></ol>

Desenvolvemos um método para formar conchas de hidrato de metano em gotículas de água sessile com segurança e compartilhar este método para máquina e montar uma célula de pressão avaliada em 10 MPa de pressão de trabalho, bem como os sistemas de pressurização e resfriamento. A célula de pressão é equipada com um estágio para a gotícula contendo termopares incorporados, uma janela de safira para visualizar a gotícula e um transdutor de pressão fixado no topo da célula. O sistema de resfriamento inclui e...

Hidratos de gás são gaiolas de moléculas de água ligadas a hidrogênio que capturam moléculas de gás convidado através de interações van der Waals. Os hidratos de metano formam-se sob condições de alta pressão e baixa temperatura, que ocorrem na natureza nos sedimentos subsuperficial ao longo das margens continentais, sob o permafrost ártico, e em outros corpos planetários no sistema solar1. Hidratos a gás armazenam vários milhares de gigatoneladas de carbono, com implicações importantes para o clima e a energia2. Hidratos de gás também podem ser perigosos na indústria de gás natural porque condições favoráveis para hidratantes ocorrem em gasodutos, que podem entupir os tubos levando a explosões fatais e derramamentos de óleo3.
Devido à dificuldade de estudar hidratos de gás in situ,experimentos laboratoriais são frequentemente empregados para caracterizar propriedades hidratadas e a influência de inibidores e substratos4. Esses experimentos de laboratório são realizados pelo cultivo de hidrato de gás a uma pressão elevada em células de várias formas e tamanhos. Os esforços para prevenir a formação de hidratos de gás em gasodutos levaram à descoberta de vários inibidores químicos e biológicos de hidrato de gás, incluindo proteínas anticongelantes (AFPs), surfactantes, aminoácidos e polivinilibrirrolidone (PVP)5,6. Para determinar os efeitos desses compostos nas propriedades de hidrato a gás, esses experimentos têm utilizado diversos projetos de embarcações, incluindo autoclaves, cristalizadores, reatores agitados e células de balanço, que suportam volumes de 0,2 a 106 centímetros cúbicos4.
O método de gotícula sessile utilizado aqui e em estudos anteriores7,8,9,10,11,12 envolve a formação de uma película hidrato a gás em uma gotícula sessile de água dentro de uma célula de pressão. Estes vasos são feitos de aço inoxidável e safira para acomodar pressões de até 10-20 MPa. A célula está conectada a um cilindro de gás metano. Dois desses estudos utilizaram o método de gotícula para testar os AFPs como inibidores de hidrato de gás em comparação com os inibidores de hidrato cinético comercial (KHIs), como PVP7,11. Bruusgard et al.7 se concentraram na influência morfológica dos inibidores e descobriram que as gotículas contendo AFPs tipo I têm uma superfície mais lisa e visada do que a superfície de gotícula dendrítica sem inibidores em altas forças motrizes.
Udegbunam et al.11 utilizaram um método desenvolvido para avaliar os KHIs em um estudo anterior10, que permite a análise de mecanismos de morfologia/crescimento, temperatura/pressão de vapor hidratado-líquido e cinética em função da temperatura. Jung et al. estudaram a substituição ch4-CO2 inundando a célula com CO2 depois de formar uma concha de hidratação CH4 8. Chen et al. observaram o amadurecimento de Ostwald como a casca de hidratação forma9. Espinoza et al. estudaram conchas hidratadas de CO2 em vários substratos minerais12. O método gotícula é um método relativamente simples e barato para determinar o efeito morfológico de vários compostos e substratos em hidratos a gás e requer pequenas quantidades de aditivos devido ao pequeno volume. Este artigo descreve um método para formar tais conchas hidratadas em uma gota de água usando uma célula de aço inoxidável com uma janela de safira para visualização, avaliada em até 10 MPa de pressão de trabalho.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CAMERA AND LAPTOP</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Camera Body</td>
			<td>Nikon</td>
			<td>D7200</td>
			<td>Name in Protocol: camera</td>
		</tr>
		<tr>
			<td>Camera Control Pro 2 Software</td>
			<td>Nikon</td>
			<td></td>
			<td>Name in Protocol: camera software</td>
		</tr>
		<tr>
			<td>Laptop</td>
			<td>HP Pavilion</td>
			<td>hp-pavilion-laptop-14-ce0068st</td>
			<td>Needs to be PC with plenty of storage (~ 1 Tb) 
			Name in Protocol: laptop</td>
		</tr>
		<tr>
			<td>Macrophotography Lens</td>
			<td>Nikon</td>
			<td>AF-S MICRO 105mm f/2.8G IF-ED Lens</td>
			<td>Name in Protocol: lens</td>
		</tr>
		<tr>
			<td>CONSUMABLES</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Deionized water</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: DI water</td>
		</tr>
		<tr>
			<td>Dry Ice</td>
			<td>VWR or grocery store</td>
			<td></td>
			<td>Buy just before nucleation 
			Name in Protocol: dry ice</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: ethanol</td>
		</tr>
		<tr>
			<td>Ethylene Glycol</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: ethylene glycol</td>
		</tr>
		<tr>
			<td>COOLING SYSTEM</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1/2 in. O.D. x 3/8 in. I.D. x 25 ft. Polyethylene Tubing</td>
			<td>Everbilt</td>
			<td>Model # 301844</td>
			<td>For circulating coolant from chiller to copper coils in aquarium 
			Name in Protocol: 3/8&#8221; (inner diameter) plastic tubing</td>
		</tr>
		<tr>
			<td>Circulating chiller</td>
			<td>Polyscience</td>
			<td></td>
			<td>Name in Protocol: chiller</td>
		</tr>
		<tr>
			<td>Economical Flexible Polyethylene Foam Pipe Insulation</td>
			<td>McMaster-Carr</td>
			<td>4530K162</td>
			<td>3/4&#34; thick wall; 1/2&#34; inner diameter; R Value 3; 6&#39; long 
			Name in Protocol: foam pipe insulation</td>
		</tr>
		<tr>
			<td>Plastic tubing</td>
			<td></td>
			<td></td>
			<td>use any tubing that fits the airline connection in the lab and long enough to travel from the airline connection to the front of the aquarium</td>
		</tr>
		<tr>
			<td>DATALOGGER</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Armature Multiplexer Module for 34970A/ 
			34972A, 20-Channel</td>
			<td>Keysight Technologies</td>
			<td>34901A</td>
			<td>Name in Protocol: datalogger multichannel</td>
		</tr>
		<tr>
			<td>Benchvue or Benchlink software</td>
			<td>Benchvue or Benchlink</td>
			<td></td>
			<td>Name in Protocol: temperature transducer software</td>
		</tr>
		<tr>
			<td>Data Acquisition/Switch Unit. GPIB, RS232</td>
			<td>Keysight Technologies</td>
			<td>34970A</td>
			<td>Name in Protocol: datalogger</td>
		</tr>
		<tr>
			<td>USB/GPIB interface</td>
			<td>Keysight Technologies</td>
			<td>82357B</td>
			<td>Name in Protocol: datalogger USB</td>
		</tr>
		<tr>
			<td>datalogger multichannel</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Schott Fostec -Llc 20510 Ace Fiber Optic Light Source</td>
			<td>Schott Fostec</td>
			<td>A20500</td>
			<td>3115PS-12W-B20 115 V ~AC 50/60Hz 5/4.5 W 
			Name in Protocol: light source unit</td>
		</tr>
		<tr>
			<td>Schott Fostec light source guide - single bundle</td>
			<td>Schott Fostec</td>
			<td>A08031.40</td>
			<td>Name in Protocol: fiber optic light source cable</td>
		</tr>
		<tr>
			<td>METHANE GAS AND REGULATOR</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1/4 OD in. x 20 ft. Copper Soft Refrigeration Coil</td>
			<td>Everbilt</td>
			<td>Model # D 04020PS</td>
			<td>For pressurizing ISCO pressure pump. An additional pack is needed for coolant circulation, as listed below. 
			Name in Protocol: high pressure-rated 1/4&#8221; copper pipe</td>
		</tr>
		<tr>
			<td>Methane cylinder regulator</td>
			<td>Airgas</td>
			<td>Y11N114G350-AG</td>
			<td>Name in Protocol: methane cylinder regulator</td>
		</tr>
		<tr>
			<td>Methane gas cylinder</td>
			<td>Airgas</td>
			<td>ME UHP300</td>
			<td>Name in Protocol: methane gas cylinder</td>
		</tr>
		<tr>
			<td>PRESSURE PUMP</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1/4 in.&#160; flexible tubing, ~ 3 ft.</td>
			<td></td>
			<td></td>
			<td>Connect to pump inlet for leak test 
			Name in Protocol: 1/4&#34;&#160; flexible tubing</td>
		</tr>
		<tr>
			<td>260D Syringe Pump W/Controller</td>
			<td>Teledyne Instruments Inc.</td>
			<td>67-1240-520</td>
			<td>Name in Protocol: pressure pump</td>
		</tr>
		<tr>
			<td>Controller &#8722; Ethernet/USB</td>
			<td>Teledyne Instruments Inc.</td>
			<td>62-1240-114</td>
			<td>Purchase if you would like to install Labview onto computer and control pressure pump remotely. We did not do this.</td>
		</tr>
		<tr>
			<td>Smooth-Bore Seamless 316 Stainless Steel Tubing, 1/4&#34; OD, 0.035&#34; Wall Thickness, 1 Foot Long (x5)</td>
			<td>McMaster-Carr</td>
			<td>89785K824</td>
			<td>Name in Protocol: 1/4&#34; pipe</td>
		</tr>
		<tr>
			<td>Smooth-Bore Seamless 316 Stainless Steel Tubing, 1/8&#34; OD, 0.02&#34; Wall Thickness, 1 Foot Long (x4)</td>
			<td>McMaster-Carr</td>
			<td>89785K811</td>
			<td>Name in Protocol: 1/8&#34; pipe</td>
		</tr>
		<tr>
			<td>Stainless Steel Swagelok Tube Fitting, Reducing Union, 1/4 in. x 1/8 in. Tube OD (x4)</td>
			<td>Swagelok</td>
			<td>&#160;SS-400-6-2</td>
			<td>Name in Protocol: 1/8&#8221; to 1/4&#8221; adapter</td>
		</tr>
		<tr>
			<td>PRESSURE CELL</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>316 Stainless Steel Nut and Ferrule Set (1 Nut/1 Front Ferrule/1 Back Ferrule) for 1/4 in. Tube Fitting (20)</td>
			<td>Swagelok</td>
			<td>&#160;SS-400-NFSET</td>
			<td>Used for fitting connections where necessary 
			Name in Protocol: ferrule set</td>
		</tr>
		<tr>
			<td>316L Stainless Steel Convoluted (FM) Hose, 1/4 in., 316L Stainless Steel Braid, 1/4 in. Tube Adapters, 60 in. (1.5 m) Length</td>
			<td>Swagelok</td>
			<td>SS-FM4TA4TA4-60</td>
			<td>Connects pressure pump to pressure cell 
			Name in Protocol: 1/4&#34; braided stainless steel flexible pressure-rated hose</td>
		</tr>
		<tr>
			<td>ABAQUS</td>
			<td>ABAQUS FEA</td>
			<td></td>
			<td>Name in Protocol: simulation software</td>
		</tr>
		<tr>
			<td>Abrasion-Resistant Cushioning Washer for 7/8&#34; Screw Size, 0.875&#34; ID, 2.25&#34; OD, packs of 10 (x1)</td>
			<td>McMaster-Carr</td>
			<td>90131A107</td>
			<td>Name in Protocol: 2.25&#34; rubber washer</td>
		</tr>
		<tr>
			<td>Abrasion-Resistant Sealing Washer, Aramid Fabric/Buna-N Rubber, 3/8&#34; Screw Size, 0.625&#34; OD, packs of 10 (x1)</td>
			<td>McMaster-Carr</td>
			<td>93303A105</td>
			<td>Used for illumination port</td>
		</tr>
		<tr>
			<td>Acrylic Sheet | White 2447 / WRT31 
			Extruded Paper-Masked (Translucent 55% (0.118 x 12 x 12)</td>
			<td>Interstate Plastics</td>
			<td>ACRW7EPSH</td>
			<td>Machine a circle of acrylic to fit in the inner chamber of the pressure cell to serve as the background for imaging 
			Name in Protocol: acrylic disc</td>
		</tr>
		<tr>
			<td>AutoCAD</td>
			<td>AutoCAD</td>
			<td></td>
			<td>Name in Protocol: engineering design software</td>
		</tr>
		<tr>
			<td>Conax fitting</td>
			<td>Conax Technologies</td>
			<td>311401-011</td>
			<td>TG(PTM2/)-24-A6-T, OPTIONAL 1/4&#34; NPT 
			Name in Protocol: pressure seal connector</td>
		</tr>
		<tr>
			<td>High Accuracy Oil Filled Pressure 
			Transducers/Transmitters for General 
			industrial applications (x2)</td>
			<td>Omega Engineering, Inc.</td>
			<td>PX409-3.5KGUSBH</td>
			<td>Buy two so there is a backup. 
			Name in Protocol: pressure transducer</td>
		</tr>
		<tr>
			<td>HIGH PRESSURE CHAMBER&#160; PARTS</td>
			<td>Wither Tool, Die and Manufacturing Company</td>
			<td></td>
			<td>Machining for pressure cell parts as listed in CAD drawings (Figure S1) 
			Name in Protocol: Part B = stainless steel washer</td>
		</tr>
		<tr>
			<td>High-Strength 316 Stainless Steel Socket Head Screw, M5 x 0.80 mm Thread, 14 mm Long (x20)</td>
			<td>McMaster-Carr</td>
			<td>90037A119</td>
			<td>Used for illumination port</td>
		</tr>
		<tr>
			<td>High-Strength 316 Stainless Steel Socket Head Screw, M8 x 1.25 mm Thread, 25 mm Long (x20)</td>
			<td>McMaster-Carr</td>
			<td>90037A133</td>
			<td>Name in Protocol: M8 stainless steel screws</td>
		</tr>
		<tr>
			<td>Oil-Resistant Hard Buna-N O-Ring, 3/32 Fractional Width, Dash Number 120, packs of 50 (x1)</td>
			<td>McMaster-Carr</td>
			<td>5308T178</td>
			<td>Name in Protocol: 1&#34; o-ring</td>
		</tr>
		<tr>
			<td>Oil-Resistant Hard Buna-N O-Ring, 3/32 Fractional Width, Dash Number 128, packs of 50 (x1)</td>
			<td>McMaster-Carr</td>
			<td>5308T186</td>
			<td>Name in Protocol: 1.5&#34; o-ring</td>
		</tr>
		<tr>
			<td>Omega Inc. pressure transducer software</td>
			<td>Omega Engineering, Inc.</td>
			<td></td>
			<td>Name in Protocol: pressure transducer software</td>
		</tr>
		<tr>
			<td>Polycarbonate Disc</td>
			<td>McMaster-Carr</td>
			<td>8571K31</td>
			<td>Listed in CAD drawings for illumination port, Fig. S1 Part E</td>
		</tr>
		<tr>
			<td>Sapphire windows (x3)</td>
			<td>Guild Optical Associates, Inc.</td>
			<td>Optical Grade Sapphire Window, C-Plane 
			Diameter: 1.811&#8221; &#177;.005&#8221; 
			Thickness: .590&#8221; &#177;.005&#8221; 
			Surface Quality: 60/40 
			Edges ground and safety chamfered</td>
			<td>Buy three so there are two backups. 
			Name in Protocol: sapphire window</td>
		</tr>
		<tr>
			<td>Solid Thermocouple Wire FEP Insulation and Jacket, Type K, 24 Gauge, 50 ft. Length (x1)</td>
			<td>McMaster-Carr</td>
			<td>3870K32</td>
			<td>Name in Protocol: thermocouples</td>
		</tr>
		<tr>
			<td>Stainless Steel Integral Bonnet Needle Valve, 0.37 Cv, 1/4 in. Swagelok Tube Fitting, Regulating Stem (x4)</td>
			<td>Swagelok</td>
			<td>&#160;SS-1RS4</td>
			<td>Two will be used for the pressure pump as well. 
			Name in Protocol: 1/4&#34; needle valves</td>
		</tr>
		<tr>
			<td>Stainless Steel Pipe Fitting, Hex Nipple, 1/4 in. Male NPT (x2)</td>
			<td>Swagelok</td>
			<td>&#160;SS-4-HN</td>
			<td>Used for illumination port</td>
		</tr>
		<tr>
			<td>Stainless Steel Swagelok Tube Fitting, Female Branch Tee, 1/4 in. Tube OD x 1/4 in. Tube OD x 1/4 in. Female NPT (x2)</td>
			<td>Swagelok</td>
			<td>&#160;SS-400-3-4TTF</td>
			<td>Used with pressure transducer 
			Name in Protocol: branch tee fitting</td>
		</tr>
		<tr>
			<td>Stainless Steel Swagelok Tube Fitting, Male Connector, 1/4 in. Tube OD x 1/4 in. Male NPT (x4)</td>
			<td>Swagelok</td>
			<td>&#160;SS-400-1-4</td>
			<td>Used on top port and side port leading to needle valves 
			Name in Protocol: NPT screws</td>
		</tr>
		<tr>
			<td>Stainless Steel Swagelok Tube Fitting, Port Connector, 1/4 in. Tube OD (x8)</td>
			<td>Swagelok</td>
			<td>&#160;SS-401-PC</td>
			<td>Use as tube connections between NTP and valve connections 
			Name in Protocol: port connector fitting</td>
		</tr>
		<tr>
			<td>TANK</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1/4 OD in. x 20 ft. Copper Soft Refrigeration Coil</td>
			<td>Everbilt</td>
			<td>Model # D 04020PS</td>
			<td>For circulating coolant 
			Name in Protocol: 1/4&#34; copper pipe</td>
		</tr>
		<tr>
			<td>10 gallon aquarium</td>
			<td>Tetra</td>
			<td></td>
			<td>Name in Protocol: 10 gallon tank</td>
		</tr>
		<tr>
			<td>2 oz. Waterweld</td>
			<td>J-B Weld</td>
			<td>Model # 8277</td>
			<td>Name in Protocol: underwater sealant</td>
		</tr>
		<tr>
			<td>3 in. x 25 ft. Foil Backed Fiberglass Pipe Wrap Insulation</td>
			<td>Frost King</td>
			<td>Model # SP42X/16</td>
			<td>For wrapping around aquarium 
			Name in Protocol: foil-lined fiberglass</td>
		</tr>
		<tr>
			<td>3/8 7/8 in. Stainless Steel Hose Clamp (10 pack)</td>
			<td>Everbilt</td>
			<td>Model # 670655E</td>
			<td>Name in Protocol: worm drive hose clamps</td>
		</tr>
		<tr>
			<td>Styrofoam</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: insulating material</td>
		</tr>
		<tr>
			<td>TOOLS</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1-1/8 in. Ratcheting Tube Cutter</td>
			<td>Husky</td>
			<td>Model # 86-036-0111</td>
			<td></td>
		</tr>
		<tr>
			<td>1/2 in. to 1 in. Pipe Cutter</td>
			<td>Apollo</td>
			<td>Model # 69PTKC001</td>
			<td></td>
		</tr>
		<tr>
			<td>Adjustable wrench (x2)</td>
			<td>Steel Core</td>
			<td>Model # 31899</td>
			<td>Need two wrenches with jaw at least 1&#34;</td>
		</tr>
		<tr>
			<td>Allen wrench set</td>
			<td>Home Depot</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Duct tape</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: duct tape</td>
		</tr>
		<tr>
			<td>Flexible tubing, like an IV line, to fit on the end of grainger probe (canula)</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: IV tube</td>
		</tr>
		<tr>
			<td>Grainger 18 gauge probe</td>
			<td>Grainger</td>
			<td></td>
			<td>For inserting droplet 
			Name in Protocol: cannula</td>
		</tr>
		<tr>
			<td>High Vacuum Grease</td>
			<td>Dow corning</td>
			<td></td>
			<td>Apply to o-rings before inserting sapphire window 
			Name in Protocol: vacuum grease</td>
		</tr>
		<tr>
			<td>Klein Tools Professional 90 Degree 4-in-1 Tube Bender</td>
			<td>Klein Tools</td>
			<td>Model # 89030</td>
			<td>Name in Protocol: tube bender</td>
		</tr>
		<tr>
			<td>Snoop liquid leak detector</td>
			<td>Swagelok</td>
			<td>MS-SNOOP-8OZ</td>
			<td>To detect leaks when pressurized when methane 
			Name in Protocol: liquid leak detector</td>
		</tr>
		<tr>
			<td>Suction cup</td>
			<td>Home Depot</td>
			<td></td>
			<td>For removing tight fitting sapphire window 
			Name in Protocol: suction cup</td>
		</tr>
		<tr>
			<td>Teflon Tape</td>
			<td></td>
			<td></td>
			<td>Name in Protocol: plumber&#39;s tape</td>
		</tr>
		<tr>
			<td>Temflex 3/4 in. x 60 ft. 1700 Electrical Tape Black</td>
			<td>3M</td>
			<td>Model # 1700-1PK-BB40</td>
			<td>Name in Protocol: electrical tape</td>
		</tr>
	</tbody>
</table>

methane hydrate crystallization on sessile water droplets

Este artigo descreve um método para formar conchas de hidrato de metano em gotículas de água. Além disso, fornece plantas para uma célula de pressão avaliada em 10 MPa de pressão de trabalho, contendo um estágio para gotículas sessile, uma janela de safira para visualização e transdutores de temperatura e pressão. Uma bomba de pressão conectada a um cilindro de gás metano é usada para pressurizar a célula a 5 MPa. O sistema de resfriamento é um tanque de 37,85 L contendo uma solução de 50% de etanol resfriada através de glicol de etileno através de bobinas de cobre. Esta configuração permite a observação da mudança de temperatura associada à formação e dissociação do hidrato durante o resfriamento e despressurização, respectivamente, bem como a visualização e fotografia das alterações morfológicas da gotícula. Com este método, observou-se formação rápida de conchas hidratantes a ~-6 °C a -9 °C. Durante a despressurização, observou-se uma queda de temperatura de 0,2 °C a 0,5 °C na curva de estabilidade de pressão/temperatura (P/T) devido à dissociação do hidratado exotérmico, confirmada pela observação visual do derretimento no início da queda da temperatura. O "efeito memória" foi observado após repressurização para 5 MPa de 2 MPa. Este projeto experimental permite o monitoramento da pressão, temperatura e morfologia da gotícula ao longo do tempo, tornando este um método adequado para testar vários aditivos e substratos na morfologia hidratada.

Com este método, uma casca de hidrato a gás em uma gotícula pode ser monitorada visualmente através de uma janela de safira da célula de pressão e através de transdutores de temperatura e pressão. Para nuclear a casca de hidratação depois de pressurizar para 5 MPa, o gelo seco pode ser adicionado ao topo da célula de pressão para induzir um choque térmico para desencadear a cristalização rápida do hidrato. Há uma clara diferença morfológica na formação de conchas de hidrato forçadas ao gelo seco. A ...

Watch this Scientific Journal Video about Methane Hydrate Crystallization on Sessile Water Droplets at JoVE.com

Methane Hydrate Crystallization on Sessile Water Droplets

Descrevemos um método para formar hidrato de gás em gotículas de água sessile para estudar os efeitos de vários inibidores, promotores e substratos na morfologia cristalina hidrato.

Cristalização do hidrato de metano em gotículas de água sessile

This paper describes a method to form methane hydrate shells on water droplets. In addition, it provides blueprints for a pressure cell rated to 10 MPa ...

O efeito de vários aditivos na morfologia do hidrato a gás e na estabilidade da temperatura de pressão pode ser testado. Mas o principal interesse está em descobrir como as biomoléculas podem interagir e afetar os hidratos gasosos in situ. A principal vantagem deste protocolo é que se pode formar uma casca hidratante em uma gotícula sessile com segurança.Nivelar a etapa de gotícula é um desafio e precisa de prática à medida que a gota desliza em um palco desigual. Além disso, familiarize-se com as conexões Swagelok e a segurança em relação a gases inflamáveis de alta pressão. Comece conectando o regulador do cilindro de metano à bomba com um tubo de cobre de uma por quatro polegadas usando um novo conjunto de porcas e ferrule.Cole uma ponta flexível de tubos iv, corte um ângulo, até o final da cânula para ajudar a direcionar a gota em direção à janela de safira. Coloque uma seringa de um mililitro na cânula e puxe o volume desejado de água deionizada. Sem a válvula da agulha ou a janela de safira anexada, insira a extremidade da cânula na placa superior e pratique a expulsão da gota para o centro do palco.Recoloque a janela de safira e as arruelas com parafusos M8 e conecte a válvula de célula de pressão superior. Conecte a mangueira de aço inoxidável trançada da bomba de pressão à célula de pressão. E verifique se todas as conexões do cilindro de gás para a célula de pressão estão apertadas.Abra a válvula de entrada da célula de pressão e coloque a célula de pressão no aquário. Em seguida, insira um cabo de fonte de luz de fibra óptica na porta de iluminação da célula de pressão. Em um aquário cheio de uma relação 50/50 de etanol e água, adicione mais etanol à medida que o nível da solução cai nas semanas seguintes, até que esteja nivelado com a parte superior da célula de pressão, logo abaixo da conexão de fonte de luz.Coloque o refrigerador à temperatura que alcançará aproximadamente zero a três graus Celsius dentro da célula e comece a circular através de bobinas. Ligue o fluxo de ar para a frente do aquário para evitar a condensação na superfície do aquário. Inicie um registro de temperatura no software de data logger.Defina o intervalo de varredura para 30 segundos e espere até que a temperatura dentro da célula de pressão esteja estável a dois graus Celsius. Depois que a câmera for colocada, ligue a fonte de luz para aproximadamente 80% e abra o software da câmera. No Live View, concentre a lente da câmera na câmara interna da célula e ajuste a fonte de luz para melhor imagem.Inicie um novo registro de temperatura com um intervalo de varredura de um segundo. Se estiver preso, desprende a válvula da agulha de saída na porta superior da célula de pressão. Coloque uma seringa de um mililitro na cânula e puxe o volume desejado de água deionizada.Insira a cânula através da placa superior até que a ponta esteja visível no software da câmera no modo Live View. Expulse a gota de fluido da seringa sobre o termopar central. Em seguida, recoloque a válvula da agulha.Concentre a câmera na gota na célula de pressão. Comece a imagem de lapso de tempo a cada 60 segundos. Abra o software do transdutor de pressão no laptop.Comece a coletar dados no gráfico e no registro de dados no intervalo de digitalização de um segundo, e espere até que a temperatura da gotícula esteja estável entre zero a três graus Celsius. Ligue a bomba e o controlador. Feche a válvula de entrada da bomba de pressão.Abra a válvula de saída da bomba e as válvulas da célula de pressão. Aque a pressão da bomba pressionando zero no controlador da bomba de pressão. Selecione a bomba A no controlador da bomba de pressão para monitorar a pressão.Certifique-se de que a bomba de pressão está vazia se um fluido diferente do gás metano estivesse presente na bomba. Faça isso definindo o fluxo máximo e o fluxo constante para 100 mililitros por minuto e pressionando Run. Deixe-o funcionando até que a bomba esteja vazia.Feche a válvula de saída da bomba e abra a válvula de entrada da bomba. Abra o cilindro de gás e coloque o regulador do cilindro de gás em 1.000 quilopascals. Pressione a recarga no controlador da bomba de pressão.Quando a bomba estiver cheia e perto de 1.000 quilopascals, feche a válvula de entrada da bomba e o cilindro de gás. Abra ligeiramente a válvula de saída da bomba para a célula. Monitore a pressão da célula de pressão no software do transdutor de pressão, pois pode diminuir devido à temperatura relativamente menor na célula de pressão.Defina o fluxo máximo para 10 mililitros por minuto, pressão máxima para 5.000 quilopascals e pressão constante para 1.000 quilopascals no controlador da bomba de pressão, conforme descrito no manuscrito do texto. Pressione Run. Quando 1.000 quilopascals for atingido, pressione Stop no controlador da bomba e feche a válvula de saída da bomba.Monitore a pressão na célula de pressão para garantir que não haja vazamentos. Se a pressão cair, use o detector de vazamento de líquido para encontrar o vazamento nas conexões e aperte cuidadosamente os componentes de vazamento. Se a célula estiver estável, abra a saída da bomba e ajuste a pressão constante gradualmente para 2.000, 3.000, 4.000 e 5.000 quilopascals, monitorando a estabilidade da célula pressionando Stop após cada ajuste.Se a pressão estiver estável, feche a saída da bomba e espere cerca de 12 a 24 horas para que o gás permeie a gota. Mude o lapso de tempo para tirar imagens a cada dois ou cinco segundos. Adicione gelo seco ao topo da célula até que a casca de hidratação seja vista em lapso de tempo.Se o gelo seco deslizar, afixe fita ao redor da parte superior da célula. Observe o progresso da formação de hidrato de metano através de fotos de lapso de tempo por duas a seis horas. Despressurize a célula para 2.000 quilopascals abrindo a saída da bomba e definindo a pressão constante para 2.000 quilopascals, observando quando ocorre o derretimento.Após 30 minutos, repressurize a célula de pressão para 5.000 quilopascals para observar o efeito da memória. Observe quando uma casca hidratante começar a se reformar e deixe a casca se formar por 30 minutos a duas horas. Despressurize a célula abrindo a saída da bomba e colocando a pressão constante em 100 quilopascals.Se houver pressão residual na célula de pressão, abra ligeiramente a válvula superior da célula de pressão em 1/16 polegadas. Salve os dados de pressão e temperatura como arquivos CSV. Remova a gotícula removendo a válvula da célula de pressão superior e extraindo a gotícula com a seringa, cânula ou tubo IV.Siga as instruções do manuscrito do texto se houver preocupação com a contaminação entre os ensaios. Há uma clara diferença morfológica sobre a formação de conchas de hidrato forçado ao gelo seco, onde a gota d'água passou de uma superfície reflexiva lisa para uma concha hidratante opaca com uma leve superfície dendrítica. A adição de 100 microgramas por mililitro nas proteínas anticongelante tipo um alterou a morfologia do hidrato induzindo bordas cristais ao longo da gotícula e saliências do topo da gotícula.Depois que a casca de hidratação se desenvolveu por uma hora, a célula foi despressurizada para dois megapascals, levando a uma queda de 2 a 5 graus Celsius na temperatura perto da curva de estabilidade P/T. Devido à dissociação do hidratado exoérmico, a dissociação do hidratado foi confirmada pelo derretimento visual através de imagens de lapso de tempo no início da diminuição da temperatura. Controles negativos sem gotícula e com gotícula que não formaram uma concha hidratante não apresentaram queda na temperatura durante a despressurização.Como o ápice de cada grau de temperatura estava acima da curva de estabilidade P/T previamente estabelecida, uma curva de regressão foi calculada com base no ápice P/T desses ensaios. Duas coisas são importantes para ter em mente ao executar este protocolo. Primeiro, verifique se todas as conexões estão seguras antes de pressurizar, e duas, pratique a expulsão da gota antes de anexar a janela de safira.Seguindo este protocolo, pode-se calcular o consumo de gás, bem como analisar as imagens para calcular a espessura da casca hidratante, para determinar o volume de hidratado formado.

methane-hydrate-crystallization-on-sessile-water-droplets

Research

JoVE Journal

Environment

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) loading from lake sediments can account for a substantial portion of the total P load in eutrophic, and some mesotrophic, lakes. Laboratory determination of P release rates from sediment cores is one approach for determining the role of internal P loading and guiding management decisions. Two principal alternatives to experimental determination of sediment P release exist for estimating internal load: in situ measurements of changes in hypolimnetic P over time and P mass balance. The experimental approach using laboratory-based sediment incubations to quantify internal P load is a direct method, making it a valuable tool for lake management and restoration.
Laboratory incubations of sediment cores can help determine the relative importance of internal vs. external P loads, as well as be used to answer a variety of lake management and research questions. We illustrate the use of sediment core incubations to assess the effectiveness of an aluminum sulfate (alum) treatment for reducing sediment P release. Other research questions that can be investigated using this approach include the effects of sediment resuspension and bioturbation on P release.
The approach also has limitations. Assumptions must be made with respect to: extrapolating results from sediment cores to the entire lake; deciding over what time periods to measure nutrient release; and addressing possible core tube artifacts. A comprehensive dissolved oxygen monitoring strategy to assess temporal and spatial redox status in the lake provides greater confidence in annual P loads estimated from sediment core incubations.

Lake eutrophication is a water quality issue worldwide, making the need to identify and control nutrient sources critical. Laboratory determination of phosphorus release rates from sediment cores is a valuable approach for determining the role of internal phosphorus loading and guiding management decisions.

Determinados pelo Laboratório Fósforo Flux do Lago Sedimentos como medida de Fósforo Interno Carregando

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

Ambiente

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

EPA Método 1615. Medição de Enterovirus e Norovirus Ocorrência na água pela cultura e RT-qPCR. I. Recolha de amostras de vírus

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a standardized method for use in multiple analytical laboratories during monitoring period 3 of the Unregulated Contaminant Monitoring Rule. Herein we present the protocol for extraction of viral ribonucleic acid (RNA) from water sample concentrates and for quantitatively measuring enterovirus and norovirus concentrations using reverse transcription-quantitative PCR (RT-qPCR). Virus concentrations for the molecular assay are calculated in terms of genomic copies of viral RNA per liter based upon a standard curve. The method uses a number of quality controls to increase data quality and to reduce interlaboratory and intralaboratory variation. The method has been evaluated by examining virus recovery from ground and reagent grade waters seeded with poliovirus type 3 and murine norovirus as a surrogate for human noroviruses. Mean poliovirus recoveries were 20% in groundwaters and 44% in reagent grade water. Mean murine norovirus recoveries with the RT-qPCR assay were 30% in groundwaters and 4% in reagent grade water.

Here we present a procedure to quantify enterovirus and norovirus in environmental and drinking waters using reverse transcription-quantitative PCR. Mean virus recovery from groundwater with this standardized procedure from EPA Method 1615 was 20% for poliovirus and 30% for murine norovirus.

EPA Método 1615. Medição de Enterovirus e Norovirus Ocorrência na água por cultura e RT-qPCR. Parte III. Detecção de vírus por RT-qPCR

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

A CO<sub&gt; 2</sub&gt; Facility Concentração Gradiente para Teste CO<sub&gt; 2</sub&gt; Enriquecimento do solo e efeitos sobre a função da pastagem de Ecossistemas

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

Protocol for Measuring the Thermal Properties of a Supercooled Synthetic Sand-water-gas-methane Hydrate Sample

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future energy resources and energy storage media. To develop a method for gas production from natural MH-bearing sediments and hydrate-based technologies, it is imperative to understand the thermal properties of gas hydrates.
The thermal properties' measurements of samples comprising sand, water, methane, and MH are difficult because the melting heat of MH may affect the measurements. To solve this problem, we performed thermal properties' measurements at supercooled conditions during MH formation. The measurement protocol, calculation method of the saturation change, and tips for thermal constants' analysis of the sample using transient plane source techniques are described here.
The effect of the formation heat of MH on measurement is very small because the gas hydrate formation rate is very slow. This measurement method can be applied to the thermal properties of the gas hydrate-water-guest gas system, which contains hydrogen, CO2, and ozone hydrates, because the characteristic low formation rate of gas hydrate is not unique to MH. The key point of this method is the low rate of phase transition of the target material. Hence, this method may be applied to other materials having low phase-transition rates.

We present a protocol for measuring the thermal properties of synthetic hydrate-bearing sediment samples comprising sand, water, methane, and methane hydrate.

Protocolo para medir as propriedades térmicas de um sintético areia-água-gás-hidrato de metano Amostra Supercooled

Methane hydrates (MHs) are present in large amounts in the ocean floor and permafrost regions. Methane and hydrogen hydrates are being studied as future ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled Earth-system processes. Such knowledge is imperative in improving predictions of hydro-biogeochemical cycles, especially under climate change scenarios. We present an experimental method, designed to capture sub-surface heterogeneity of an initially homogeneous soil system. This method is based on destructive sampling of a soil lysimeter designed to simulate a small-scale hillslope. A weighing lysimeter of one cubic meter capacity was divided into sections (voxels) and was excavated layer-by-layer, with sub samples being collected from each voxel. The excavation procedure was aimed at detecting the incipient heterogeneity of the system by focusing on the spatial assessment of hydrological, geochemical, and microbiological properties of the soil. Representative results of a few physicochemical variables tested show the development of heterogeneity. Additional work to test interactions between hydrological, geochemical, and microbiological signatures is planned to interpret the observed patterns. Our study also demonstrates the possibility of carrying out similar excavations in order to observe and quantify different aspects of soil-development under varying environmental conditions and scale.

This study presents an excavation method for investigating subsurface hydrological, geochemical, and microbiological heterogeneity of a soil lysimeter. The lysimeter simulates an artificial hillslope which was initially under homogeneous condition and had been subjected to approximately 5,000 mm of water over eight cycles of irrigation in an 18-month period.

Monólito escavação para Coupled Hidrológicos, geoquímicos e microbiológicos Investigações

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

Protocolo para Microplásticos Amostragem na superfície do mar e análise de amostras

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

A Simple Planting Technique for Re-establishing Trees Where Frequent Inundation Occurs

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with herbaceous plants. This protocol describes a simple, cost-effective tree planting technique needed for restoring degraded Everglade tree islands. The design is patterned after a natural Everglades process that creates floating peat islands, which allows tree survival and growth in flooded conditions and often leads to the development of tree islands. Commercially available peat bags were used as the medium for the growth and establishment of potted native tree saplings. The pop-up configuration floated initially and provided additional elevation to minimize inundation, with a single native tree species sapling and a single tree fertilizer spike. During a 3 year study involving 105 pop-ups, most plants survived (80%) and many thrived. Determining whether this technique can establish trees on a degraded tree island will require longer studies and extensive field tests.

This protocol describes a simple, cost-effective tree planting technique for restoring degraded Everglades tree islands that experience inundation. The design creates an island (pop-up) which floats initially and adds elevation to promote tree survival and growth under flooded conditions.

Uma simples técnica de plantação para restabelecer as árvores onde ocorre a inundação frequente

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with ...

Experimental Study of the Relationship Between Particle Size and Methane Sorption Capacity in Shale

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard for evaluating the mining value of shale gas. Currently, studies on the correlation between particle size and methane adsorption are controversial. In this study, an isothermal adsorption apparatus, the gravimetric sorption analyzer, is used to test the adsorption capacity of different particle sizes in shale to determine the relationship between the particle size and the adsorption capacity of shale. Thegravimetric method requires fewer parameters and produces better results in terms of accuracy and consistency than methods like the volumetric method. Gravimetric measurements are performed in four steps: a blank measurement, preprocessing, a buoyancy measurement, and adsorption and desorption measurements. Gravimetric measurement is presently considered to be a more scientific and accurate method of measuring the amount of adsorption; however, it is time-consuming and requires a strict measuring technique. A Magnetic Suspension Balance (MSB) is the key to verify the accuracy and consistency of this method. Our results show that adsorption capacity and particle size are correlated, but not a linear correlation, and the adsorptions in particles sieved into 40 - 60 and 60 - 80 meshes tend to be larger. We propose that the maximum adsorption corresponding to the particle size is approximately 250 &#181;m (60 mesh) in the shale gas fracturing.

We use an isothermal adsorption apparatus, the gravimetric sorption analyzer, to test the adsorption capacity of different particle sizes of shale, in order to find out the relationship between particle size and the adsorption capacity of shale.

Estudo experimental da relação entre o tamanho de partícula e capacidade de adsorção de metano em xisto

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard ...

Construction of a Low-cost Mobile Incubator for Field and Laboratory Use

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water quality. However, commercially available incubators are often costly, difficult to transport, not flexible in terms of volume, and/or poorly adapted to local field conditions where access to electricity is unreliable. The purpose of this study was to develop an adaptable, low-cost and transportable incubator that can be constructed using readily available components. The electronic core of the incubator was first developed. These components were then tested under a range of ambient temperature conditions (3.5 &#176;C - 39 &#176;C) using three types of incubator shells (polystyrene foam box, hard cooler box, and cardboard box covered with a survival blanket). The electronic core showed comparable performance to a standard laboratory incubator in terms of the time required to reach the set temperature, inner temperature stability and spatial dispersion, power consumption, and microbial growth. The incubator set-ups were also effective at moderate and low ambient temperatures (between 3.5 &#176;C and 27 &#176;C), and at high temperatures (39 &#176;C) when the incubator set temperature was higher. This incubator prototype is low-cost (&lt; 300 USD) and adaptable to a variety of materials and volumes. Its demountable structure makes it easy to transport. It can be used in both established laboratories with grid power or in remote settings powered by solar energy or a car battery. It is particularly useful as an equipment option for field laboratories in areas with limited access to resources for water quality monitoring.

This paper describes a method for building an adaptable, low-cost and transportable incubator for microbial testing of drinking water. Our design is based on widely available materials and can operate under a range of field conditions, while still offering the advantages of higher-end laboratory-based models.

Construção de uma incubadora móvel de baixo custo para o campo e uso em laboratório

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water ...