Name: a low-cost method of measuring the in situ primary productivity of periphyton communities of lentic waters
Uploaded: 2022-12-16T13:05:00
Description: Watch this Scientific Journal Video about A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters at JoVE.com

This study was supported by the Czech Science Foundation (GACR 19-05791S), RVO 67985939, and by the CAS within the program of the Strategy AV 21, Land save and recovery. Many thanks to Ond&#345;ej Sihelsk&#253; for taking the shots in the field - without him, the filming would have been complete hell. The project would not be possible without tight cooperation with companies, Palivov&#253; Kombin&#225;t &#218;st&#237; s.p. and Sokolovsk&#225; Uheln&#225;, who provided access to the studied localities.

NOTE: Before sampling, determine the degree of replications based on the overall project needs, statistical design, or expected amount of sample variability.Five replicate pairs of light and dark incubation bottles are suggested for precise statistical analysis and to account for potential sample loss or breakage. The described floating experimental barge is designed to carry five replicates plus one pair of blank controls; see Figure 1 for a technical drawing of the experimental barge.
...

<ol>
	<li>Blachart, J. L., 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=Potential+consequences+of+climate+change+for+primary+production+and+fish+production+in+large+marine+ecosystems.">Potential consequences of climate change for primary production and fish production in large marine ecosystems.</a> Philosophical Transactions of the Royal Society B: Biological Sciences. 367 (1605), 2979-2989 (2012).</li><li>Howarth, R. W., Michaels, A. F., Sala, O. E., Jackson, R. B., Mooney, H. A., Howarth, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Measurement+of+primary+production+in+aquatic+ecosystems.">The Measurement of primary production in aquatic ecosystems.</a> Methods in Ecosystem Science. , 72-85 (2000).</li><li>Vadenbecouer, Y. E. G., Peterson, M. J., Vander, Z., Kalff, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Benthic+algal+production+across+lake+size+gradients:+Interactions+among+morphometry,+nutrients,+and+light.">Benthic algal production across lake size gradients: Interactions among morphometry, nutrients, and light.</a> Ecology. 89 (9), 2542-2552 (2008).</li><li>Reimer, A., Landmann, G., Kempe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lake+Van,+eastern+Anatolia,+hydrochemistry+and+history.">Lake Van, eastern Anatolia, hydrochemistry and history.</a> Aquatic Geochemistry. 15 (1), 195-222 (2009).</li><li>Cantonati, M., Lowe, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lake+benthic+algae:+toward+an+understanding+of+their+ecology.">Lake benthic algae: toward an understanding of their ecology.</a> Freshwater Sciences. 33 (2), 475-486 (2014).</li><li>Gaarder, T., Gran, H. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+production+of+plankton+in+the+Oslo+Fjord.+Rapports+et+Proces-verbaux+des+R&#233;unions.">Investigation of the production of plankton in the Oslo Fjord. Rapports et Proces-verbaux des R&#233;unions.</a> Conseil International pour l'&#201;xploration de la Mer. 42, 1-48 (1927).</li><li>Hall, R. O., Thomas, S., Gaiser, E. E., Fahey, T. J., Knapp, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+Freshwater+Primary+Productivity+and+Respiration.">Measuring Freshwater Primary Productivity and Respiration.</a> Principles and Standards for Measuring Primary Productivity. , (2007).</li><li>Howart, R., Michaels, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+6+The+Measurement+of+Primary+Production+in+Aquatic+Ecosystems.">Chapter 6 The Measurement of Primary Production in Aquatic Ecosystems.</a> Springer Science and Business Media LLC. , (2000).</li><li>Kop&#225;&#269;ek, J., Hejzlar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Semi-micro+determination+of+total+phosphorus+in+soils,+sediments,+and+organic+materials:+a+simplified+perchloric+acid+digestion+procedure.">Semi-micro determination of total phosphorus in soils, sediments, and organic materials: a simplified perchloric acid digestion procedure.</a> Communications in Soil Science and Plant Analysis. 26 (11-12), 1935-1946 (1995).</li><li>Benson, B. B., Krause, 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+concentration+and+isotopic+fractionation+of+oxygen+dissolved+in+freshwater+and+seawater+in+equilibrium+with+the+atmosphere1.">The concentration and isotopic fractionation of oxygen dissolved in freshwater and seawater in equilibrium with the atmosphere1.</a> Limnology and Oceanography. 29 (3), 620-632 (1984).</li><li>Dodds, W. K., Biggs, B. J., Lowe, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photosynthesis-irradiance+patterns+in+benthic+microalgae:+variations+as+a+function+of+assemblage+thickness+and+community+structure.">Photosynthesis-irradiance patterns in benthic microalgae: variations as a function of assemblage thickness and community structure.</a> Journal of Phycology. 35 (1), 42-53 (1999).</li><li>Bott, T. L., 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=An+evaluation+of+techniques+for+measuring+periphyton+metabolism+in+chambers.">An evaluation of techniques for measuring periphyton metabolism in chambers.</a> Canadian Journal of Fisheries and Aquatic Sciences. 54 (3), 715-725 (1997).</li><li>Blankenship, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+and+functional+dynamics+of+photosynthetic+antenna+complexes.">Structural and functional dynamics of photosynthetic antenna complexes.</a> Proceedings of the National Academy of Sciences. 112 (45), 13751-13752 (2015).</li><li>Hawes, I., Schwartz, A. -. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photosynthesis+in+an+extreme+shade+environment,+benthic+microbial+mats+from+Lake+Hoare,+a+permanently+ice-covered+Antarctic+lake.">Photosynthesis in an extreme shade environment, benthic microbial mats from Lake Hoare, a permanently ice-covered Antarctic lake.</a> Journal of Phycology. 35 (3), 448-459 (1999).</li><li>Aristegui, J., 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=Planktonic+primary+production+and+microbial+respiration+measured+by+14C+assimilation+and+dissolved+oxygen+changes+in+coastal+waters+of+the+Antarctic+peninsula+during+austral+summer:+Implications+for+carbon+flux+studies.">Planktonic primary production and microbial respiration measured by 14C assimilation and dissolved oxygen changes in coastal waters of the Antarctic peninsula during austral summer: Implications for carbon flux studies.</a> Marine Ecology-Progress Series. 132, 191-201 (1996).</li><li>Steemann-Nielsen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+radioactive+carbon+(14C)+for+measuring+organic+production+in+the+sea.">The use of radioactive carbon (14C) for measuring organic production in the sea.</a> Journal of Experimental Marine Biology and Ecology. 18 (2), 117-140 (1952).</li><li>Sanz-Mart&#237;n, 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=Relationship+between+carbon-and+oxygen-based+primary+productivity+in+the+Arctic+Ocean,+svalbard+archipelago.">Relationship between carbon-and oxygen-based primary productivity in the Arctic Ocean, svalbard archipelago.</a> Frontiers in Marine Science. 6, 468 (2019).</li><li>Nielsen, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+the+production+of+organic+matter+in+the+sea+by+means+of+carbon-14.">Measurement of the production of organic matter in the sea by means of carbon-14.</a> Nature. 167 (4252), 684-685 (1951).</li><li>J&#246;nsson, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+14C-incubation+technique+for+measuring+microphytobenthic+primary+productivity+in+intact+sediment+cores.">A 14C-incubation technique for measuring microphytobenthic primary productivity in intact sediment cores.</a> Limnology and Oceanography. 36 (7), 1485-1492 (1991).</li><li>Bender, M. L., 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=A+comparison+of+four+methods+for+determining+planktonic+community+production.">A comparison of four methods for determining planktonic community production.</a> Limnology and Oceanography. 32 (5), 1085-1098 (1987).</li><li>&#352;imek, 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=Spatio-temporal+patterns+of+bacterioplankton+productivity+and+community+composition+related+to+phytoplankton+composition+and+protistan+bacterivory+in+a+dam+reservoir.">Spatio-temporal patterns of bacterioplankton productivity and community composition related to phytoplankton composition and protistan bacterivory in a dam reservoir.</a> Aquatic Microbial Ecology. 51 (3), 249-262 (2008).</li></ol>

The methodology described in this paper is based on the principle of the light and dark bottle oxygen technique in combination with the noninvasive technique of measuring O2 concentration using optical oxygen sensors. This system allows the parallel measurement of different incubation settings as the optical fiber for measuring O2 can be moved quickly from bottle to bottle. The benthic communities from various depths can differ in taxonomic composition and productivity; simultaneously measuring them...

Primary productivity is the only entry of autochthonous carbon into the aquatic systems that form the entire system food web1. Hence, the accurate estimation of primary productivity is an essential step toward understanding the functioning of aquatic ecosystems. Littoral zones are areas of high primary productivity and biodiversity. In addition to phytoplankton, periphyton (hereafter referred to as microbial mats) and macroalgae are assumed to significantly contribute to primary productivity in littoral zones2. Due to their sessile lifestyle and significant spatial heterogeneity, quantification of primary productivity is not trivial. 
Primary productivity is driven mainly by light intensity, temperature, availability of nutrients, and distribution of the ionic species of the carbonate system in the respective depths of euphotic zones3,4. The depth markedly influences the spatial distribution of microbial mats. Microbial communities must cope with the adverse effects of high irradiation and pronounced seasonal temperature variations in shallow depths and with lower light intensity at greater depths. In addition to the depth gradient, dynamic trophic interactions generate multiple and complex spatial patterns at different scales5. This complex system is complicated to simulate in the laboratory. The most accurate way to infer the metabolic activity of individual primary producers from littoral zones is to set up in situ experiments. 
The methodology introduced in this paper is based on the traditional chamber method2,6,7, together with a transportable and easy-to-build low-cost floating barge. This allows the measurement of primary productivity at different depths under the natural light spectrum, temperature, and different distribution of the ionic species of the carbonate system with the depth. The method is based on the principle of light versus dark bottle oxygen, which was first employed to measure phytoplankton photosynthesis6 and is still commonly used6,7. It compares the rate of change in oxygen in bottles kept in the light (which includes the effects of primary productivity and respiration) with those held in the dark (respiration only)8. The method uses oxygen evolution (photosynthesis) as a proxy for primary productivity. The measured variables are net ecosystem productivity (NEP, as a change in O2 concentration over time in light conditions) and ecosystem respiration (RE, as a change in O2 concentration over time in the dark). Gross ecosystem productivity (GEP) is the calculation of the difference between the two (Table 1). The term "ecosystem" is used here to denote that the periphyton is composed of autotrophic and heterotrophic organisms. The most significant improvement of this traditional chamber method is using noninvasive oxygen optical sensors and optimization of this primarily planktonic method for measuring periphytic primary productivity.
The technique is described in the example of measuring microbial mats in the littoral zone of newly emerged post-mining lakes in the Czech Republic-Milada, Most, and Medar. The metabolic activity of microbial mats is determined using direct in situ measurement of O2 fluxes performed directly at specific depths, where the studied communities naturally occur. Heterotrophic and phototrophic activity is measured in closed glass bottles equipped with noninvasive optical oxygen sensors. These sensors detect the partial pressure of oxygen using the fluorescence of light-sensitive dyes. The bottles with microbial mats are suspended and incubated on a floating device at the appropriate depths. The oxygen concentration inside the bottles was continuously measured during the daylight period from the small boat. 
Samples of intact microbial mats are collected and placed in gas-tight incubation bottles at designated depths by scuba divers. Each bottle is equipped with a noninvasive optical oxygen microsensor, which monitors the O2 productivity/consumption over time. All measurements are done in five replicate dark/light pairs in each depth. The temperature and photosynthetically active radiation (PHAR) intensities are measured at respective depths throughout the incubation. After 6 h of in situ incubation (daylight hours), the microbial mats are harvested from the bottles and dried. O2 fluxes are normalized to microbial biomass. As a control, fluxes are corrected for changes in O2 concentration in separate light and dark gas-tight bottles (blank controls) containing lake water without microbial mat biomass. Below are detailed instructions for building the floating barge and performing the whole experiment step-by-step. This paper also presents representative results from the measurements of microbial mats at two depths (1 m and 2 m), with five replicates at each depth. Actual temperature and light intensity were measured during the whole experiment using dataloggers.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Aluminum angle L profile 40 x 40 mm x 3 mm, length 2,000 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Aluminum flat bar 40 x 3 x 350 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bucket 15 L with concrete infill&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Carabine hook with screw lock 50 x 5 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>electric tape black</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Extruded polystyrene (XPS) material 500 x 200 x 150 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fibox 3 LCD trace</td>
			<td>PreSens Precision Sensing GmbH</td>
			<td></td>
			<td>stand-alone fiber optic oxygen meter</td>
		</tr>
		<tr>
			<td>Hondex PS-7 Portable Depth Sounder</td>
			<td>Hondex&#160; - Honda Electronics</td>
			<td></td>
			<td>to measures distances through water - to bottom depth measurement; https://www.honda-el.net/industry/ps-7e</td>
		</tr>
		<tr>
			<td>KORKEN - glass tight-seal jar 0.5 L</td>
			<td>IKEA</td>
			<td></td>
			<td>incubation bottles; https://www.ikea.com/cz/en/p/korken-jar-with-lid-clear-glass-70213545/</td>
		</tr>
		<tr>
			<td>metal hook&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen Sensor Spot SP-PSt3-NAU-D5</td>
			<td>PreSens Precision Sensing GmbH</td>
			<td></td>
			<td>non-invasive optical oxygen sensor for measurements under Real Conditions</td>
		</tr>
		<tr>
			<td>SCOUT infantable canoe</td>
			<td>GUMOTEX</td>
			<td></td>
			<td>https://www.gumotexboats.com/en/scout-standard#0000-044667-021-13/11C</td>
		</tr>
		<tr>
			<td>Screw 10 x 170 mm with hexagonal nuts</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Screw 4 x 15 mm with hexagonal nuts</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Screw 4 x 15 mm with wing nuts</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Snap hooks 50 x 5 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Steel Carabine hook 50 x 5 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Steel chain with wire diameter 3 mm, inside link 5.5 x 26 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Steel chain, 5 m</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>toothbrush</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>tweezer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Washer 10 x 50 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Washer 4 x 10 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Washer 4 x 10 mm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a low-cost method of measuring the in situ primary productivity of periphyton communities of lentic waters

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers (mainly phosphorus concentration and light intensity) and species composition on primary productivity. Primary productivity is mainly driven by light intensity, temperature, availability of nutrients, and distribution of the ionic species of the carbonate system in the respective depths of the euphotic zone. It is a complex system that is very difficult to simulate in the laboratory. This cheap, transportable, and easy-to-build floating barge allows measuring the primary productivity accurately-directly under the actual natural conditions. The methodology is based on measuring the primary productivity in real time using noninvasive oxygen sensors integrated into tightly sealed glass jars, enabling online oxygen flux monitoring and providing new insights into metabolic activities. Detailed seasonal in situ measurements of gross primary productivity of microbial mats (or other benthic organisms) can improve current knowledge of the processes controlling primary productivity dynamics in lentic waters.

<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/64078/64078fig05.jpg" /> 
Figure 5: Net and gross ecosystem productivity of microbial mats during daylight. (A) Light bottle-net ecosystem productivity: time course data of net oxygen productivity of microbial mats from the light bottles. The oxygen concentration change in incubation bottles was measured after 1 h during daylight. Grey circles: bottles with samples of m...

Watch this Scientific Journal Video about A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters at JoVE.com

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Presented here is a cost-effective and transportable method/facility for measuring the primary productivity of microbial mats under actual in situ environmental temperature and light conditions. The experimental setup is based on widely available materials and can be used under various conditions while offering the advantages of laboratory-based models.

A Low-Cost Method of Measuring the In Situ Primary Productivity of Periphyton Communities of Lentic Waters

Measuring the in situ primary productivity of periphyton during the growing season gradient can elucidate the quantitative effect of environmental drivers ...

Research

JoVE Journal

Environment

Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays

Much of the nutrient cycling and carbon  processing in natural environments occurs through the activity of extracellular  enzymes released by ...

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

Quantification of Heavy Metals and Other Inorganic Contaminants on the Productivity of Microalgae

Increasing demand for renewable fuels has researchers investigating the feasibility of alternative feedstocks, such as microalgae. Inherent advantages ...

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 ...

Pretreatment of Lignocellulosic Biomass with Low-cost Ionic Liquids

A number of ionic liquids (ILs) with economically attractive production costs have recently received growing interest as media for the delignification of ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus (Murayama), Using a Low-Cost, Small, and Easily Constructed Flight Mill

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). ...

A Flexible Low Cost Hydroponic System for Assessing Plant Responses to Small Molecules in Sterile Conditions

A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for ...

Construction of a Compact Low-Cost Radiation Shield for Air-Temperature Sensors in Ecological Field Studies

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although ...

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 ...