I would like to acknowledge Kelpie Wilson of Wilson Biochar, the Utah Bureau of Land Management, the USDA Forest Service, the Utah Division of Forestry, Fire and State Lands, the Utah Public Lands Initiative Grant Program, the Utah State University Extension Grant Program, Brandon Barron of Burns, OR,&#160;ANR Fabrication of Logan, Utah, and the U.S. Biochar Initiative.

1.&#160;Transportation to site
<ol>
	<li>Load the BB12 onto a trailer for transport using either a mini-excavator (with thumb) and chains OR a 4 x 4 forklift, utilizing the fork pockets. 
	NOTE: All further references to the mini-excavator assume it has an operable thumb with the bucket.</li>
	<li>Secure the kiln to the trailer with ratchet straps with a breaking strength of over 1,361 kilograms (3,000 lbs.) each. See Figure 4 for an example.</li>
	<li>Unload the kiln using the mini-excavator or forklift or&#160;tie it to a tree/vehicle and drive away, allowing it to fall on the ground. Once at the site, drag the kiln on the ground on its skids using a pickup truck and straps.</li>
</ol>
2.&#160;On-site preparation
<ol>
	<li>Place the BB12 on a relatively flat surface (slopes &#60; 10%) fairly close to the pile of wood, ensuring the dog-door draining panel is on the downhill side, if on a slope. 
	NOTE: Keep the dog-door draining panel closed during operation.</li>
	<li>Select feedstocks, which can be any woody species. Use small, dry material to get the kiln going, especially when working with wet feedstock.</li>
	<li>Gather and pile an adequate amount of feedstock to feed the kiln. Cut all the feedstock to a length that is shorter than the maximum dimension of the kiln so it fits in easily.</li>
</ol>
3.&#160;Hazard reduction
<ol>
	<li>Before ignition, check with local fire and air quality authorities to ensure that the fire danger is not too high for ignition and that the air quality regulations do not impact burning plans. Be sure to alert local fire authorities of plans before ignition.</li>
	<li>Before ignition, build a fire line (1 foot-wide trail scraped down to mineral soil) entirely around the kiln to prevent a fire from creeping away. Ensure there is a charged hose connected to an adequate water supply, dependent on the current fire hazard, for fire control before igniting the kiln.</li>
</ol>
4.&#160;Loading and lighting kiln
<ol>
	<li>Load the kiln randomly with feedstock using the mini-excavator; the size/order of adding the feedstocks does not matter. See Figure 5 for an image of a loaded&#160;kiln.</li>
	<li>Pile a layer of smaller-diameter fuels (feedstock) on top of the kiln to aid in quick ignition.</li>
	<li>Once loaded, light the kiln with a drip torch, propane torch, or other ignition device by lighting the top of the kiln first. See Figure 6 for images of lighting the kiln and kiln operation.</li>
</ol>
5.&#160;Tending kiln
<ol>
	<li>Allow the kiln to burn down and add fuels as the fire allows; expect a short period of smoke production after materials are added to the kiln. Observe that the flame cap will soon reform and continue to consume the combustibles as they rise through the heat/smoke column. Tend the kiln similar to a campfire; adding too much fuel at once will smother the fire, but too little will cause the fire to go out. See Figure 7 for an image of a flame cap.</li>
	<li>Monitor the kiln and surrounding vegetation for escaped sparks or embers that can cause unwanted ignition.</li>
	<li>Continue tending/loading the kiln this way until the kiln is full/ the feedstock is depleted/the end of the shift.</li>
</ol>
6.&#160;Quenching
<ol>
	<li>When the kiln is full or nearly full of coals and the flaming combustion gives way to smoldering or glowing combustion, quench to halt combustion and preserve the coals. Figure 8 shows a kiln ready to quench.</li>
	<li>Use a 3.8 cm (1.5 inch) diameter hose and water pump (often referred to as volume pump or trash pump at rental stores) from a water source or water truck to quench the kiln using approximately 300 gallons of water. Drown the coals; continue adding water until the coals are in standing water and stir using the mini-excavator to eliminate dry hot spots and continued combustion. Figure 9 demonstrates quenching with a hose.</li>
	<li>Once the kiln is fully quenched, open the dog door to drain out the water, which will make the kiln lighter for tipping out the biochar.</li>
</ol>
7.&#160;Tipping
<ol>
	<li>Once the kiln is completely quenched and drained, dump the contents onto the ground adjacent to the kiln.</li>
	<li>Prepare the area first by using the machine to construct a fire line around the space where the coals will be, and scrape away any fuels, including grass, sticks, brush, and logs from this space.</li>
	<li>Use the machine and chains/straps to pull the kiln over towards the machine. Figure 10 shows the kiln being tipped by the mini-excavator.</li>
	<li>Monitor the boom/hydraulic hoses/fittings of the mini-excavator for heat exposure; occasionally place the back of a hand first near and eventually on these parts to ensure they are not too hot to touch; if so, allow them to cool immediately.</li>
</ol>
8.&#160;Cold trailing
<ol>
	<li>Before leaving the site, ensure the fire is out by moving hands through the entire amount of biochar that was produced. Make sure that it is completely cool to the touch to call the fire out. 
	NOTE: The BB12 comes with lids that can be used for safety purposes; if an emergency were to occur, such as an escaped fire or injury, the lids can be placed on the kiln to prevent sparks/embers from exiting the kiln and the area can be evacuated safely. The procedure can be stalled at any time. In high-humidity locations where fire danger is generally low, lids may not be necessary.</li>
</ol>

Operating a Large Biochar Kiln and Implementing Best Practices

<ol>
	<li>Hardy, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wildland+fire+hazard+and+risk:+Problems,+definitions,+and+context.">Wildland fire hazard and risk: Problems, definitions, and context.</a> Forest Ecology and Management. 211 (1-2), 73-82 (2005).</li><li>Wollstein, K., O'Connor, C., Gear, J., Hoagland, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimize+the+bad+days:+Wildland+fire+response+and+suppression+success.">Minimize the bad days: Wildland fire response and suppression success.</a> Rangelands. 44 (3), 187-193 (2022).</li><li>Hessburg, P., Reynolds, K., Keane, R., James, K., Salter, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+wildland+fire+danger+and+prioritizing+vegetation+and+fuels+treatments.">Evaluating wildland fire danger and prioritizing vegetation and fuels treatments.</a> Forest Ecology and Management. 247 (1-3), 1-17 (2007).</li><li>Busse, M. D., Shestak, C. J., Hubbert, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soil+heating+during+burning+of+forest+slash+piles+and+wood+piles.">Soil heating during burning of forest slash piles and wood piles.</a> International Journal of Wildland Fire. 22 (6), 786-796 (2013).</li><li>Galinato, S. P., Yoder, J. K., Granatstein, 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+economic+value+of+biochar+in+crop+production+and+carbon+sequestration.">The economic value of biochar in crop production and carbon sequestration.</a> Energy Policy. 39 (10), 6344-6350 (2011).</li><li>Spokas, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+the+stability+of+biochar+in+soils:+predictability+of+O:C+molar+ratios.">Review of the stability of biochar in soils: predictability of O:C molar ratios.</a> Carbon Management. 1 (2), 289 (2010).</li><li>Duku, M. H., Gu, S., Hagan, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochar+production+potential+in+Ghana-A+review.">Biochar production potential in Ghana-A review.</a> Renewable and Sustainable Energy Reviews. 15 (8), 3539-3551 (2011).</li><li>Inoue, Y., Mogi, K., Yoshizawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+cinders+from+red+pine,+black+locust,+and+henon+bamboo.">Properties of cinders from red pine, black locust, and henon bamboo.</a> Asia Pacific Biochar Conference, APBC Kyoto. , (2011).</li><li>Cornelissen, G., 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=Emissions+and+char+quality+of+flame-curtain+&amp;#34;Kon-Tiki&amp;#34;+kilns+for+farmer-scale+charcoal/biochar+production.">Emissions and char quality of flame-curtain &amp;#34;Kon-Tiki&amp;#34; kilns for farmer-scale charcoal/biochar production.</a> PLoS ONE. 11 (5), e0154617 (2016).</li><li>Puettmann, M., Kamalakanta, S., Wilson, K., Oneil, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+cycle+assessments+of+biochar+produced+from+forest+residues+using+portable+systems.">Life cycle assessments of biochar produced from forest residues using portable systems.</a> Journal of Cleaner Production. 250 (2020), 119564 (2020).</li><li>. Utah Biomass Resources Website Available from: <a target="_blank" href="https://www.usu.edu/ubrg/biochar/simple-kiln-technology">https://www.usu.edu/ubrg/biochar/simple-kiln-technology</a> (2017)</li><li>Adam, 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=Improved+and+more+environmentally+friendly+charcoal+production+system+using+a+low-cost+retort-kiln+(Eco-charcoal).">Improved and more environmentally friendly charcoal production system using a low-cost retort-kiln (Eco-charcoal).</a> Renewable Energy. 34 (8), 1923-1925 (2009).</li><li>Biomass to biochar: Maximizing the carbon value. Pullman, WA: Washington State University, Center for Sustaining Agriculture and Natural Resources Available from: <a target="_blank" href="https://csanr.wsu.edu/biomass2biochar/">https://csanr.wsu.edu/biomass2biochar/</a> (2021)</li></ol>

The author has no conflict of interest to declare.

Typically, a portion of the biochar produced on the site is collected by workshop participants in buckets or bags and applied to people&#39;s gardens or agricultural projects. Biochar is friable and can be broken into small pieces to more readily incorporate into the soil by driving over it with a vehicle, stepping on it with a hard surface beneath, or mashing it with the bucket of the mini-excavator. This material can also be referred to as charcoal and has been collected for outdoor charcoal cooking, potentially providing a locally sourced material to add to the culinary features of a meal.
Comparing Big Box flame cap biochar kilns to other biochar production methods12, mobile carbonizers can process 63,502 kg per day (70 tons), compared with 12,500 kg per day with a Big Box kiln. The cost of mobile carbonizers is much higher than a Big Box kiln, starting at $500,000 to purchase, as opposed to less than $10,000 to have a Big Box kiln manufactured. Although a single Big Box kiln can process only 20% of the material that a mobile carbonizer can, it will cost only 2% of the purchase price of a mobile carbonizer.
Heated auger kilns can process up to 5,443 kg of biomass per day, as another example, which is much lower than the 12,500 kg per day capacity of Big Box kilns. Additionally, the cost of preprocessing (chipping) the material can be more than actually pyrolyzing the material. Furthermore, refined machines such as the heated auger will not tolerate soiled feedstock that is common in forestry operations; a shovelful of soil can shut down an auger kiln while a Big Box kiln can tolerate several shovelfuls of soil without significantly impacting the operation. Finally, the cost of an auger kiln can easily be 10 times that of a Big Box kiln.
The first Big Box kiln constructed is referred to as the BB16 as it measures 4.9 m (16 ft) long by 2.4 m (8 ft) wide and is a single-walled construction. It was originally 1.8m (6 ft) high and weighed close to 1,360 kg (3,000 lbs.), which required a larger excavator, a qualified operator, and a low-boy trailer, which led to scheduling challenges. This approach was oversized to deal with the typical Utah fuel loads, and at 1.8 m (6 ft) tall, it was very difficult to light or see what was going on down inside the kiln. To address these issues, to better scale this approach to Utah fuel loads, and to make it more accessible to the average forest manager, the height was reduced to 1.2 m (4 ft) tall. This makes it easier to see into and to ignite. It also got it down to 1,043 kg (2,300 lbs.), which made it manageable to transport with a more available pickup truck and trailer, and to move and operate with a mini-excavator that requires no previous experience and can be rented from most equipment rental shops.
The second kiln built by the UBRG is a double-walled construction, which allows for better heat protection to the operators and equipment near the kiln and allows for more even heating inside the kiln13. Part of this modification was to go from 12-guage steel to 14-gauge steel, which is thinner and lighter. The UBRG has done dozens of burns in these kilns and while they do get a little bent up in spots, they are not yet showing any obvious signs of heat-related metal fatigue. Certainly, additional learning is likely to occur and there is ample room for continued innovation.
The double-walled BB12 is the design that has garnered the most attention and is perhaps the most accessible/practical for fuels in the Intermountain West. Larger kilns will be more appropriate with more/larger fuels, such as the Northwest US. This method has been proven up to a 4.9 m (16-ft-long kiln). To date, Big Box kilns have been built by other parties in Utah, Colorado, Montana, Texas, and New York.
The kilns can be operated to maximize biochar production or maximize hazardous fuels reduction, or somewhere between. If hazardous fuel reduction is the primary goal, then the kilns can be loaded randomly and quenched only when the kiln is full of coals. If surrounding fire danger is low, such as when the ground is covered in several centimeters of snow, the kilns can be stacked high with fuels in the evening before the end of the shift and left to burn all night; thus, consuming fuels in a controlled space. If biochar production is the primary goal, the feedstock can be sorted into like sizes and the kilns loaded with similar size class material and quenched frequently to preserve the coals. Typically, it is a mixture of these opposing goals and the kilns are operated between these two extremes. The species of the feedstock is less important unless a biochar with specific properties is the goal.
A limited amount of smoke comes out of these kilns; the idea is that the flame cap consumes the combustibles as they rise through the column of heat. In 2019 and 2020, the Utah Smoke Management System Coordinator Paul Corrigan brought his emissions testing equipment to Big Box biochar kiln demonstrations near Logan, in northern Utah, and Moab, in southern Utah. In both cases, the equipment registered no increase in emissions from the kilns because the flame-cap consumes the combustibles as they rise through the heat column. In April of 2023, the USDA Forest Service Fire Lab&#39;s emissions testing team conduct emissions testing on the kilns in Tooele, Utah; these results are not yet available.
The workers tending the kiln will need firefighting hand tools such as shovels, rakes, Pulaskis, and chainsaws. Best practices include having all people present wear safety gear such as leather gloves, eye protection, fire-resistant clothing or at least natural fiber clothing; synthetic clothing must be avoided. Hard hats and leather boots, long sleeves, and pants help to protect operators.
Emergency communication; contingency planning: The location (often remote) of the operation must be considered in addition to the possibility of an emergency and the communication needs around that. It is crucial to know where local cell phone reception might work best; a satellite phone or emergency locator beacon such as a Garmin InReach would be highly recommended. It is important not to work alone.
In case of an escaped ember/spot fire, the lid must be placed on the kiln to prevent further sparks from exiting the kiln. The machinery must be used to quickly dig a fire line around the spot fire, and the burning fuels must be separated from the unburned fuels. The water source must be used to extinguish the fire. If extinguishment cannot be obtained immediately, call 911.
Biochar from Big Box kilns has been characterized by Control Laboratories in Watsonville, CA using the International Biochar Initiative (IBI) Laboratory Tests for Certification Program and the results show 85% Organic Carbon and 8% ash; these are the characteristics of a moderately high-quality biochar. Collaborators are experimenting with adding roll-off coasters on the bottom, similar to large garbage dumpsters, as well as a door that is one of the end-walls to aid in removing the finished biochar. It remains to be seen if these features remain operable after extreme heat exposure.

Hazardous fuels are a major problem on wildlands throughout the West1. As fire managers can do little to control the weather, controlling fuels is their best option2. The goal of this method is to provide a new, scalable tool for reducing waste wood while producing biochar in an economically and practically accessible manner. Foresters traditionally pile and burn material from logging and fuel reduction projects, but air quality restrictions and longer fire seasons have made open pile burning much more difficult over the past decades3. Furthermore, open pile burning has been shown to cause potential long-term damage to soils from the excessive heat4. All these challenges provide the reason why the UBRG is developing this technique for biochar production. The UBRG set out to provide a low-cost, high-accessibility approach to hazardous fuel reduction that results in a valuable product5. This approach of turning fuels into feedstocks and attempting to make value from low-value wood is rife with challenges. This approach conserves a portion of that carbon, which is otherwise lost to burning or rotting, and processes it into a durable form, with a half-life approaching 1,000 years in the soil6; this is 10-1,000 times longer than residence times of most soil organic matter7.
The Big Box kiln design process began with a review of other derivatives of a technology that originated in Japan. In 2011, Inoue et al.8 reported on the carbonizing efficiency and quality of biochar produced in the &#34;smokeless charcoal kiln M50&#34; manufactured by the Moki company in Japan. Biochar was produced in these small, cone-shaped kilns with conversion efficiencies ranging between 13% and 19.5% on a dry mass basis. The authors found that the values of fixed carbon and carbon content of the char were equal to those of charcoal made in a retort method at the temperature of about 600 &#176;C.
The Big Box shape was first suggested by Kelpie Wilson in a feasibility study for the North Dakota Forest Service on carbonizing shelterbelt tree removals. Wilson suggested using a modified steel trash dumpster as a flame cap kiln for processing larger-sized material. The Big Box kiln design includes several improvements to the concept that help with durability, usability, and mobility, as described below. Wilson&#39;s figure includes suggestions for repurposing containers such as dumpsters and oil tanks for this purpose; however, repurposed material generally has been painted or galvanized and may expose workshop participants to harmful chemicals in the air.
Emissions from the Big Box Kiln have not yet been reported, but Cornelissen et al.9 conducted emissions testing on several different types of Kon-Tiki kilns (a deep cone kiln) and found that emissions were generally lower than those from open burning of biomass feedstocks. They also tested the biochars produced for polycyclic aromatic hydrocarbons (PAH) and found that PAH levels were well below the Norwegian maximum tolerable risk (MTR) level for soils. A life-cycle analysis of the Oregon Kiln (a shallow pyramid-shaped kiln) showed that the in-woods operation of a flame cap kiln was carbon-negative, resulting in a net sequestration of atmospheric carbon in soil10.
One limitation of the Big Box approach is wet feedstock. While two batches per day of large-diameter material in these kilns is a reasonable expectation in arid climates and dry feedstock, one batch a day is a more reasonable expectation in locations with higher humidity and fuel moisture. Dry feedstock is more productive; wet feedstock will limit the productivity of the kiln. Wet feedstock on a wet day does not work well. Wet feedstock less than 10 cm in diameter will pyrolyze more completely than larger-diameter wet feedstock. Dry material can easily be pyrolyzed in wet and/or snowy weather. Big Box kilns have successfully pyrolyzed kiln-length dry logs upwards of 0.76 m (30 inches) in diameter and branches down to less than 1 cm in diameter.
Kiln operation is treated like open pile burning by most air quality regulators and, in Utah, permission is only granted three&#160;days in advance, which makes planning difficult, especially in winter months when atmospheric inversions are common around our communities. The cost of conducting a biochar burn is much higher than simply burning the piles, which presents another limitation of this approach. This technique is the first published low-tech method for biochar production at a scale that forgoes expensive preprocessing of feedstocks such as grinding and chipping prior to pyrolyzing. This method is useful for most woody debris that has not been chipped or processed beyond cutting into manageable piece sizes. This method is not useful for small piece-size feedstocks or feedstocks that will form mats or globs of material such as grasses, corn stover, and rice hulls.
Kiln design 
The BB12 is a double-walled kiln that is 3.7 m (12 feet) long, 1.8 m (6 feet) wide, and 1.2 m (4 feet tall), made from 14-gauge steel. The size and shape can be varied. Plans are available on the UBRG Website11. No air is allowed into the kiln except at the top; this is critical to developing the flame cap that consumes most of the combustibles as they rise through the heat column. See Figure 1 for details of the inside corners of the kiln. The exception is a drainage port, referred to as the dog door, shown in Figure 2 because it is similar in size to a common dog door. It has a sliding piece of metal with a handle so it can be pushed down closed when operating the kiln and lifted (Caution: Hot) when ready to dump the kiln.
The two walls are separated to provide an air gap12 and are open on the top and not completely sealed on the bottom, except on the interior of the kiln. See Figure 3 for details of the air gap and the top of walls. Avoid sealed spaces to avoid problems during heat expansion and resultant contraction. Single-walled kilns are still effective at reducing hazardous fuels and producing biochar, but the double-walled kiln allows for equipment and operators to get close with less heat exposure. If biochar production is the most important goal, a double-walled kiln might be more effective. If hazardous fuels reduction is the primary goal and biochar is secondary, a single-walled kiln is likely adequate.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Big Box biochar kiln</td>
			<td>ANR Fabrication</td>
			<td>BB12</td>
			<td>Phone: 435-753-0310</td>
		</tr>
		<tr>
			<td>Chainsaw</td>
			<td>Ace Hardware</td>
			<td>#7000565</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Drip torch</td>
			<td>Forestry Suppliers</td>
			<td>7.58182E+11</td>
			<td>Fill with: 30% gasoline, 70% diesel (or propane torch)</td>
		</tr>
		<tr>
			<td>Garmin InReach</td>
			<td>Best Buy</td>
			<td>6499326</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Honda self priming water pump</td>
			<td>Northern Tool</td>
			<td>Item#&#160;109418</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>lighter / matches</td>
			<td>Amazon</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Log chains</td>
			<td>Tractor Supply Co.</td>
			<td>SKU: 358788199</td>
			<td>Or similar; for moving/lifting kiln</td>
		</tr>
		<tr>
			<td>Mini-excavator with thumb</td>
			<td>Local rental company</td>
			<td></td>
			<td>Must have a bucket with thumb</td>
		</tr>
		<tr>
			<td>Pulaski</td>
			<td>Grainger</td>
			<td>485C27</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Rachet straps</td>
			<td>US Cargo Control</td>
			<td></td>
			<td>3,000 lbs strength per</td>
		</tr>
		<tr>
			<td>Rags / cardboard</td>
			<td>Grainger</td>
			<td>9JZ92</td>
			<td>To protect straps from abrasion while transporting kiln</td>
		</tr>
		<tr>
			<td>Rake</td>
			<td>Grainger</td>
			<td>1WG30</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Shovel</td>
			<td>Grainger</td>
			<td>12N166</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Truck / trailer</td>
			<td>Own/rent locally</td>
			<td></td>
			<td>For transporting kiln</td>
		</tr>
		<tr>
			<td>Water discharge hoses</td>
			<td>Grainger</td>
			<td>45DT92</td>
			<td></td>
		</tr>
		<tr>
			<td>Water: 300 gallons per quench</td>
			<td></td>
			<td></td>
			<td>Plus more for fire control</td>
		</tr>
		<tr>
			<td>Personal Protective Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chainsaw chaps</td>
			<td>Global industrial</td>
			<td>T9FB2019133</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Ear protection</td>
			<td>Global industrial</td>
			<td>T9F708377</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Eye protection</td>
			<td>Global industrial</td>
			<td>T9F708119CLAF</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Fire pants</td>
			<td>Grainger</td>
			<td>12R487</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Fire shirt</td>
			<td>Grainger</td>
			<td>39EM96</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Hard hat</td>
			<td>Global industrial</td>
			<td>T9FB2278977</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Leather gloves</td>
			<td>Global industrial</td>
			<td>&#160;T9FB1145414</td>
			<td>Or similar</td>
		</tr>
		<tr>
			<td>Smokejumper fire boots</td>
			<td>Whites Boots</td>
			<td></td>
			<td>Or similar</td>
		</tr>
	</tbody>
</table>

big box biochar kiln operation and best practices

Big Box biochar kilns are an alternative to open pile burning that allows for in-woods biochar production in a simple metal box with no moving parts. This approach is based on technology used by charcoal makers for centuries but with a modern, mechanized approach. A mini-excavator or other piece of machinery is used to load, tend, and empty the kilns. This article will outline Big Box biochar kiln best practices, including the design, transportation, placement, loading, lighting, quenching, and dumping procedures for beginners developing their own Big Box biochar kiln programs. 
Biochar production requires a low oxygen burning environment, and the Big Box kilns use a flame-cap (sometimes referred to as a flame-curtain) method to burn material with limited smoke production. These kilns have been designed to be easily transferred to the site using an adequately rated trailer. A mini-excavator or other piece of machinery is used to load, tend, and empty the kilns. The author is not aware of a more accessible means for people to sequester durable carbon on the farm, ranch, or in the backyard. This article outlines Big Box biochar kiln best practices, including the design, transportation, placement, loading, lighting, quenching, and dumping procedures for beginners developing their own Big Box biochar kiln programs.

From October to March, Big Box kilns pyrolyzed various types of feedstocks into biochar (Table 1). The drier and cleaner the fuels are, the more productive the kilns are. The diameter of the fuels is less important, the kilns have pyrolyzed full-length logs upwards of 76 cm diameter; however, if biochar production is the most important aspect of the project, it is necessary to note that filling the kilns with more consistently sized feedstock may yield the highest output of biochar. The kilns can be operated in such a way as to maximize biochar production, or they can be operated to maximize hazardous fuel consumption, or they can be operated to focus on any point along the continuum between these somewhat opposite goals.
The lids are heavy and sharp and must not be handled alone. Figure 11 shows the lid being moved by two people. It is best to have 2-3 people tending the kiln; one operating the machine, and the others keeping a lookout for escaped spot fires, cutting any material with a chainsaw that is too long to go into the kiln, picking up small pieces of burning material that may have fallen out of the kiln.
The skids on the bottom of the kiln allow it to be dragged short distances. The kilns can be dragged at least a quarter mile, on dirt roads and over the ground. Figure 12 shows the kiln being towed. When the last part of a feedstock pile is put in the kiln, instead of completing the batch and quenching a partially full kiln, it is more efficient to drag the kiln between piles while it is operating, using a pickup truck and straps. There are eyelets near the bottom of the kiln to attach straps or chains for dragging. Then, once at a fresh pile, operators can continue loading the kiln as per directions.
There will usually be logs that do not completely pyrolyze into biochar that can be added to the next kiln burn or can be spread in the forest as coarse woody debris, which offers ecological advantages or can be used for H&#252;gelkultur applications (wood is mounded and buried to create a raised garden bed). Figure 13 shows incompletely pyrolyzed logs, sometimes called bones.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65362/65362fig01.jpg" /> 
Figure 1: Inside empty kiln. Inside of an empty kiln; note the lack of air gaps. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/65362/65362fig02.jpg" /> 
Figure 2: Open dog door. The dog door is partially opened and water is flowing out of the kiln after quenching is complete. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig02large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/65362/65362fig03.jpg" /> 
Figure 3: Gap between walls. The gap between the walls of the kiln, demonstrating no sealed cavities. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig03large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/65362/65362fig04.jpg" /> 
Figure 4: Kiln secured for transport. Strapping and trailer are used to transport the kiln safely. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig04large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/65362/65362fig05.jpg" /> 
Figure 5: Loading the kiln. The kiln is being loaded with Russian olive wood, demonstrating the loading method and lack of organization within the kiln. An example of a kiln loaded with light feedstocks. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig05large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/65362/65362fig06.jpg" /> 
Figure 6: Lighting kiln. The operator is using drip torch to top-light a loaded kiln. An example of a kiln loaded with heavy materials. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig06large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/65362/65362fig07.jpg" /> 
Figure 7: Flame cap. Flame cap formed over the top of the kiln, very little visible smoke coming from kiln; clean view to the landscape in the background. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig07large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/65362/65362fig08.jpg" /> 
Figure 8: Ready to quench. A nearly full kiln shifting from flaming combustion to glowing combustion; the point at which the quench commences. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig08large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/65362/65362fig09.jpg" /> 
Figure 9: Quench. A fire hose is being used to put approximately 1,100 liters of water on the coals in the kiln to halt combustion and preserve the char. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig09large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 10" class="xfigimg" src="/files/ftp_upload/65362/65362fig10.jpg" /> 
Figure 10: Tipping. A mini-excavator is used to tip a BB16 Big Box biochar kiln to empty the char and start another batch. Note a second kiln operating in the background; one machine can operate several kilns simultaneously. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig10large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 11" class="xfigimg" src="/files/ftp_upload/65362/65362fig11.jpg" /> 
Figure 11: Lids. Two operators wearing heavy leather gloves work together to place a lid on the kiln. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig11large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 12" class="xfigimg" src="/files/ftp_upload/65362/65362fig12.jpg" /> 
Figure 12: Towing. Straps are used to attach the (still burning) kiln from the bottom eyelets to a pickup truck and drug over a dirt road to the next pile of feedstock. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig12large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 13" class="xfigimg" src="/files/ftp_upload/65362/65362fig13.jpg" /> 
Figure 13: Bones. Incompletely pyrolyzed material, called bones, from the previous batch is added to a kiln before a new batch is started. <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/65362/65362fig13large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Big Box kilns in&#160;</td>
			<td colspan="2">Pyrolysis of&#160;</td>
			<td>Biochar&#160;</td>
		</tr>
		<tr>
			<td>Logan Ranger District of Uinta-Wasatch-Cache National Forest in November</td>
			<td>22,000 kg</td>
			<td>juniper</td>
			<td>14 cubic meters&#160;</td>
		</tr>
		<tr>
			<td>Moab, Utah in January</td>
			<td>1,200 kg</td>
			<td>Russian Olive feedstock</td>
			<td>8 cubic meters&#160;</td>
		</tr>
		<tr>
			<td>Mill Hollow in Uinta-Wsatch-Cache National Forest in November</td>
			<td>25,000 kg</td>
			<td>large-diameter Engelmann Spruce and subalpine fir</td>
			<td>16 cubic meters</td>
		</tr>
		<tr>
			<td>Near Dillon, Montana, in October</td>
			<td>10,000 kg</td>
			<td>Douglas fir feedstock</td>
			<td>10 cubic meters&#160;</td>
		</tr>
		<tr>
			<td>Pine Valley Ranch, Utah, in March</td>
			<td>24,000 kg</td>
			<td>quaking aspen, subalpine fir, Engelmann Spruce feedstock</td>
			<td>14 cubic meters&#160;</td>
		</tr>
	</tbody>
</table>
Table 1: Pyrolysis of various feedstocks by Big Box kilns.

Watch this Scientific Journal Video about Big Box Biochar Kiln Operation and Best Practices at JoVE.com

Installation of the Big Box Biochar Kiln for Biochar Production

The Utah Biomass Resources Group (UBRG) has scaled-up simple biochar kilns in an innovative approach to hazardous fuel reduction and biochar production using metal boxes, called Big Box kilns, that allow for in-woods biochar production. This article outlines Big Box biochar kiln operation and best practices.

Big Box Biochar Kiln Operation and Best Practices

Big Box biochar kilns are an alternative to open pile burning that allows for in-woods biochar production in a simple metal box with no moving parts. This ...

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2.3K Views.  Utah State University. The Utah Biomass Resources Group (UBRG) has scaled-up simple biochar kilns in an innovative approach to hazardous fuel reduction and biochar production using metal boxes, called Big Box kilns, that allow for in-woods biochar production. This article outlines Big Box biochar kiln operation and best practices.

Video: Installation of the Big Box Biochar Kiln for Biochar Production

Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling

Tracing rare stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2 fluxes and soil organic matter formation to small-scale stable-isotope biomarker probing. Coupling multiple stable isotopes such as 13C with 15N, 18O or&#160;2H has the potential to reveal even more information about complex stoichiometric relationships during biogeochemical transformations. Isotope labeled plant material has been used in various studies of litter decomposition and soil organic matter formation1-4. From these and other studies, however, it has become apparent that structural components of plant material behave differently than metabolic components (i.e. leachable low molecular weight compounds) in terms of microbial utilization and long-term carbon storage5-7. The ability to study structural and metabolic components separately provides a powerful new tool for advancing the forefront of ecosystem biogeochemical studies. Here we describe a method for producing 13C and 15N labeled plant material that is either uniformly labeled throughout the plant or differentially labeled in structural and metabolic plant components.
Here, we present the construction and operation of a continuous 13C and 15N labeling chamber that can be modified to meet various research needs. Uniformly labeled plant material is produced by continuous labeling from seedling to harvest, while differential labeling is achieved by removing the growing plants from the chamber weeks prior to harvest. Representative results from growing Andropogon gerardii Kaw demonstrate the system&#39;s ability to efficiently label plant material at the targeted levels. Through this method we have produced plant material with a 4.4 atom%13C and 6.7 atom%15N uniform plant label, or material that is differentially labeled by up to 1.29 atom%13C and 0.56 atom%15N in its metabolic and structural components (hot water extractable and hot water residual components, respectively). Challenges lie in maintaining proper temperature, humidity, CO2 concentration,&#160;and light levels in an airtight 13C-CO2 atmosphere for successful plant production. This chamber description represents a useful research tool to effectively produce uniformly or differentially multi-isotope labeled plant material for use in experiments on ecosystem biogeochemical cycling.

Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling

This method explains how to build and operate a continuous 13C and 15N isotope labeling chamber for uniform or differential plant tissue labeling. Representative results from metabolic and structural labeling of Andropogon gerardii are discussed.

Tracing rare stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2 fluxes ...

Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals is often not well understood. Here we demonstrate an integrated field lysimetry and porewater sampling method for evaluating the mobility of chemicals applied to soils and established vegetation. Lysimeters, open columns made of metal or plastic, are driven into bareground or vegetated soils. Porewater samplers, which are commercially available and use vacuum to collect percolating soil water, are installed at predetermined depths within the lysimeters. At prearranged times following chemical application to experimental plots, porewater is collected, and lysimeters, containing soil and vegetation, are exhumed. By analyzing chemical concentrations in the lysimeter soil, vegetation, and porewater, downward leaching rates, soil retention capacities, and plant uptake for the chemical of interest may be quantified. Because field lysimetry and porewater sampling are conducted under natural environmental conditions and with minimal soil disturbance, derived results project real-case scenarios and provide valuable information for chemical management. As chemicals are increasingly applied to land worldwide, the described techniques may be utilized to determine whether applied chemicals pose adverse effects to human health or the environment.

Field lysimetry and porewater sampling allow researchers to evaluate the fate of chemicals applied to soils and established vegetation. The goal of this protocol is to demonstrate how to install required instrumentation and collect samples for chemical analysis during integrated field lysimetry and porewater sampling experiments.

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals ...

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination control. Developments of some techniques aiming to reduce trace metal contamination in the last couple of decades have resulted in concentrations reported now being orders of magnitude lower than those in the past. These low concentrations often necessitate preconcentration of water samples prior to instrumental analysis of samples. Since contamination can appear in all phases of trace metal analyses, including sample collection (and during preparation of sampling containers), storage and handling, pretreatments, and instrumental analysis, specific care needs to be taken in order to reduce contamination levels at all steps. The effort to develop and utilize "clean techniques" in trace metal studies allows scientists to investigate trace metal distributions and chemical and biological behavior in greater details. This advancement also provides the required accuracy and precision of trace metal data allowing for environmental conditions to be related to trace metal concentrations in aquatic environments.
This protocol that is presented here details needed materials for sample preparation, sample collection, sample pretreatment including preconcentration, and instrumental analysis. By reducing contamination throughout all phases mentioned above for trace metal analysis, much lower detection limits and thus accuracy can be achieved. The effectiveness of "clean techniques" is further demonstrated using low field blanks and good recoveries for standard reference material. The data quality that can be obtained thus enables the assessment of trace metal distributions and their relationships to environmental parameters.

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination ...

Operation of a 25 KWth Calcium Looping Pilot-plant with High Oxygen Concentrations in the Calciner

Calcium looping (CaL) is a post-combustion CO2 capture technology that is suitable for retrofitting existing power plants. The CaL process uses limestone as a cheap and readily available CO2 sorbent. While the technology has been widely studied, there are a few available options that could be applied to make it more economically viable. One of these is to increase the oxygen concentration in the calciner to reduce or eliminate the amount of recycled gas (CO2, H2O and impurities); therefore, decreasing or removing the energy necessary to heat the recycled gas stream. Moreover, there is a resulting increase in the energy input due to the change in the combustion intensity; this energy is used to enable the endothermic calcination reaction to occur in the absence of recycled flue gases. This paper presents the operation and first results of a CaL pilot plant with 100% oxygen combustion of natural gas in the calciner. The gas coming into the carbonator was a simulated flue gas from a coal-fired power plant or cement industry. Several limestone particle size distributions are also tested to further explore the effect of this parameter on the overall performance of this operating mode. The configuration of the reactor system, the operating procedures, and the results are described in detail in this paper. The reactor showed good hydrodynamic stability and stable CO2 capture, with capture efficiencies of up to 70% with a gas mixture simulating the flue gas of a coal-fired power plant.

Operation of a 25 KWth Calcium Looping Pilot-plant with High Oxygen Concentrations in the Calciner

This manuscript describes a procedure for operating a calcium looping pilot-plant for post-combustion carbon capture with high oxygen concentrations in the calciner in order to reduce or eliminate the flue gas recycle.

Calcium looping (CaL) is a post-combustion CO2 capture technology that is suitable for retrofitting existing power plants. The CaL process uses limestone ...

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop cultivars against environmental stresses. An experimental protocol is presented for RGB and hyperspectral imaging of root systems. The approach uses rhizoboxes where plants grow in natural soil over a longer time to observe fully developed root systems. Experimental settings are exemplified for assessing rhizobox plants under water stress and studying the role of roots. An RGB imaging setup is described for cheap and quick quantification of root development over time. Hyperspectral imaging improves root segmentation from the soil background compared to RGB color based thresholding. The particular strength of hyperspectral imaging is the acquisition of chemometric information on the root-soil system for functional understanding. This is demonstrated with high resolution water content mapping. Spectral imaging however is more complex in image acquisition, processing and analysis compared to the RGB approach. A combination of both methods can optimize a comprehensive assessment of the root system. Application examples integrating root and aboveground traits are given for the context of plant phenotyping and plant physiological research. Further improvement of root imaging can be obtained by optimizing RGB image quality with better illumination using different light sources and by extension of image analysis methods to infer on root zone properties from spectral data.

An experimental protocol is presented for assessment of soil grown plant root systems with RGB and hyperspectral imaging. Combination of RGB image time series with chemometric information from hyperspectral scans optimizes insights into plant root dynamics.

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

Isolation and Analysis of Microbial Communities in Soil, Rhizosphere, and Roots in Perennial Grass Experiments

Plant and soil microbiome studies are becoming increasingly important for understanding the roles microorganisms play in agricultural productivity. The purpose of this manuscript is to provide detail on how to rapidly sample soil, rhizosphere, and endosphere of replicated field trials and analyze changes that may occur in the microbial communities due to sample type, treatment, and plant genotype. The experiment used to demonstrate these methods consists of replicated field plots containing two, pure, warm-season grasses (Panicum virgatum and Andropogon gerardii) and a low-diversity grass mixture (A. gerardii, Sorghastrum nutans, and Bouteloua curtipendula). Briefly, plants are excavated, a variety of roots are cut and placed in phosphate buffer, and then shaken to collect the rhizosphere. Roots are brought to the laboratory on ice and surface sterilized with bleach and ethanol (EtOH). The rhizosphere is filtered and concentrated by centrifugation. Excavated soil from around the root ball is placed into plastic bags and brought to the lab where a small amount of soil is taken for DNA extractions. DNA is extracted from roots, soil, and rhizosphere and then amplified with primers for the V4 region of the 16S rRNA gene. Amplicons are sequenced, then analyzed with open access bioinformatics tools. These methods allow researchers to test how the microbial community diversity and composition varies due to sample type, treatment, and plant genotype. Using these methods along with statistical models, the representative results demonstrate there are significant differences in microbial communities of roots, rhizosphere, and soil. Methods presented here provide a complete set of steps for how to collect field samples, isolate, extract, quantify, amplify, and sequence DNA, and analyze microbial community diversity and composition in replicated field trials.

Excavation of plant roots from the field as well as processing of samples into endosphere, rhizosphere, and soil are described in detail, including DNA extraction and data analysis methods. This paper is designed to enable other laboratories to use these techniques for the study of soil, endosphere, and rhizosphere microbiomes.

Plant and soil microbiome studies are becoming increasingly important for understanding the roles microorganisms play in agricultural productivity. The ...

Generating Homo- and Heterografts Between Watermelon and Bottle Gourd for the Study of Cold-responsive MicroRNAs

MicroRNAs (miRNAs) are endogenous small non-coding RNAs of about 20 - 24 nt, known to play important roles in plant development and adaptation. There is an accumulating evidence showing that the expressions of certain miRNAs are altered when grafting, an agricultural practice commonly used by farmers to improve crop tolerance to biotic and abiotic stresses. Bottle gourd is an inherently climate-resilient crop compared to many other major cucurbits, including watermelon, rendering it one of the most widely used rootstocks for the latter. The recent advancement of high-throughput sequencing technologies has provided great opportunities to investigate cold-responsive miRNAs and their contributions to heterograft advantages; yet, adequate experimental procedures are a prerequisite for this purpose. Here, we present a detailed protocol for efficiently generating homo- and heterografts between the cold-susceptible watermelon and the cold-tolerant bottle gourd, in addition to methods of tissue sampling, data generation, and data analysis. The presented methods are also useful for other plant-grafting systems, to interrogate miRNA regulations under various environmental stresses, such as heat, drought, and salinity.

Here we present a detailed protocol for efficiently making homo- and heterografts between watermelon and bottle gourd, in addition to methods of tissue sampling, data generation, and data analysis, for the investigation of cold-responsive microRNAs.

MicroRNAs (miRNAs) are endogenous small non-coding RNAs of about 20 - 24 nt, known to play important roles in plant development and adaptation. There is ...

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are delivered using a non-quantitative method. Accurate delivery of known quantities of nucleic acids or other chemicals is critical to measure the efficacy of the compound being tested and to allow reliable comparison between compounds.
Here we provide a reproducible, quantitative microinjection protocol that ensures accurate delivery of specific doses of dsRNA, reducing the mortality typically induced by injection injury. These modifications include the addition of Rhodamine B, a graduated injection needle, and an improved recovery method borrowed from Isoe and Collins. This method allows calculation of dose responses and facilitates comparisons between compounds. Versions of this method have been successfully used on three genera of mosquitoes as well as house flies to assess the reduction in fecundity resulting from gene silencing of ribosomal RNA transcripts.
This protocol provides strategies to reduce several challenges of small insect microinjection. Together, mechanical delivery of dsRNAs accompanied by visual verification, identification of effective locations for delivery, and inclusion of a post-injection recovery period ensure accurate dosing and low injury mortality. This protocol also describes an oviposition bioassay for uniform determination of effects on fecundity.

This protocol describes a microinjection methodology that we have standardized and used for several years to deliver specific quantities of nucleic acids directly to the hemolymph of mosquitoes and house flies. This protocol results in minimal injection mortality and allows dose correlated measurements of fecundity.

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are ...

Colletotrichum fioriniae Development in Water and Chloroform-based Blueberry and Cranberry Floral Extracts

To accurately monitor the phenology of the bloom period and the temporal dynamics of floral chemical cues on fungal fruit rotting pathogens, floral extraction methods and coverslip bioassays were developed utilizing Colletotrichum fioriniae. In blueberry and cranberry, this pathogen is optimally controlled by applying fungicides during the bloom period because of the role flowers play in the initial stages of infection. The protocol detailed here describes how floral extracts (FE) were obtained using water-, chloroform-, and field rainwater-based methods for later use in corresponding glass coverslip bioassays. Each FE served to provide a different set of information: response of C. fioriniae to mobilized floral chemical cues in water (water-based), pathogen response to flower and fruit surface waxes (chloroform-based), and field-based monitoring of collected floral rainwater, moving in vitro observations to an agricultural setting. The FE is broadly described as either water- or chloroform-based, with an appropriate bioassay described to compensate for the inherent differences between these two materials. Rainwater that had run off flowers was collected in unique devices for each crop, alluding to the flexibility and application of this approach for other crop systems. The bioassays are quick, inexpensive, simple, and provide the ability to generate spatiotemporal and site-specific information about the presence of stimulatory floral compounds from various sources. This information will ultimately better inform disease management strategies, as FE decrease the time needed for infection to occur, thus providing insight into changing risks for pathogen infection over the growing season.

Colletotrichum fioriniae Development in Water and Chloroform-based Blueberry and Cranberry Floral Extracts

Here, bioassays designed to monitor the development of a fungal pathogen, Colletotrichum fioriniae, in the presence of blueberry or cranberry floral extracts on glass coverslips are described. Water-, chloroform-, and field rainwater- based floral extraction techniques are detailed as well as insight into how this information can be applied.

To accurately monitor the phenology of the bloom period and the temporal dynamics of floral chemical cues on fungal fruit rotting pathogens, floral ...

Manufacturing Simple and Inexpensive Soil Surface Temperature and Gravimetric Water Content Sensors

Quantifying temperature and moisture at the soil surface is essential for understanding how soil surface biota respond to changes in the environment. However, at the soil surface these variables are highly dynamic and standard sensors do not explicitly measure temperature or moisture in the upper few millimeters of the soil profile. This paper describes methods for manufacturing simple, inexpensive sensors that simultaneously measure the temperature and moisture of the upper 5 mm of the soil surface. In addition to sensor construction, steps for quality control, as well as for calibration for various substrates, are explained. The sensors incorporate a Type E thermocouple to measure temperature and assess soil moisture by measuring the resistance between two gold-plated metal probes at the end of the sensor at a depth of 5 mm. The methods presented here can be altered to customize probes for different depths or substrates. These sensors have been effective in a variety of environments and have endured months of heavy rains in tropical forests as well as intense solar radiation in deserts of the southwestern U.S. Results demonstrate the effectiveness of these sensors for evaluating warming, drying, and freezing of the soil surface in a global change experiment.

Accurately measuring temperature and water content of the upper 5 mm of the soil surface can improve our understanding of environmental controls on biological, chemical, and physical processes. Here we describe a protocol for manufacturing, calibrating, and conducting measurements with soil surface temperature and moisture sensors.

Quantifying temperature and moisture at the soil surface is essential for understanding how soil surface biota respond to changes in the environment. ...

Big Box Biochar Kiln Operation and Best Practices

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Chapters in this video

Design and Operation of a Continuous ¹³C and ¹⁵N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling

Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Operation of a 25 KW_th Calcium Looping Pilot-plant with High Oxygen Concentrations in the Calciner

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Isolation and Analysis of Microbial Communities in Soil, Rhizosphere, and Roots in Perennial Grass Experiments

Generating Homo- and Heterografts Between Watermelon and Bottle Gourd for the Study of Cold-responsive MicroRNAs

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Colletotrichum fioriniae Development in Water and Chloroform-based Blueberry and Cranberry Floral Extracts

Manufacturing Simple and Inexpensive Soil Surface Temperature and Gravimetric Water Content Sensors