Name: dendrochronological dating and provenancing of string instruments
Uploaded: 2022-10-06T14:50:00
Description: Watch this Scientific Journal Video about Dendrochronological Dating and Provenancing of String Instruments at JoVE.com

This study was supported by the Slovenian Research Agency (ARRS) Program P4-0015 (Wood and lignocellulosic composites) and the Young Researchers&#39; Program.

1. Inspection and description of a stringed musical instrument&#160;-&#160;violin
NOTE: Violins are the most frequently investigated stringed instruments. Therefore, we describe the procedure on a violin.
<ol>
	<li>Examine the instrument and all its parts. Take detailed photos of the front (top), back, side, scroll, and label along with the measuring scale for the final report and the archive (Figure 1).</li>
	<li>Examine the top plate of...

<ol>
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Geigen- und Lautenmacher des Barock. , 266-275 (2015).</li><li>Eckstein, D., Bauch, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beitrag+zur+Rationalisierung+eines+dendrochronologischen+Verfahrens+und+zur+Analyse+seiner+Aussagesicherheit.">Beitrag zur Rationalisierung eines dendrochronologischen Verfahrens und zur Analyse seiner Aussagesicherheit.</a> Forstwissenschaftliches Centralblatt. 88, 230-250 (1969).</li><li>Baillie, M. G. L., Pilcher, J. 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The presented protocol describes the procedure of the dendrochronological dating of a violin. The procedure includes several critical steps. The first one is to identify the tree rings in order to properly measure their width. This is critical because the tree rings are often very narrow or have unclear boundaries due to the small amount of latewood (step 1.3). The detection of tree rings can be complicated by aging of the wood, dark and opaque varnishes28, or damage, repairs, retouching, or dirt ...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64591">Dendrochronological Dating and Provenancing of String Instruments</a>. The Authors section was updated from:
Katarina &#268;ufar1 
Bla&#382; Dem&#353;ar2 
Micha Beuting3 
Angela Balzano1 
Nina &#352;krk1 
Luka Kr&#382;e1 
Maks Merela1 
1Department of Wood Science and Technology, Biotechnical Faculty, University of Ljubljana 
2University of Ljubljana 
3Universitat Hamburg
to:
Katarina &#268;ufar1 
Bla&#382; Dem&#353;ar2 
Micha Beuting2 
Angela Balzano1 
Nina &#352;krk1 
Luka Kr&#382;e1 
Maks Merela1 
1Department of Wood Science and Technology, Biotechnical Faculty, University of Ljubljana 
2Independent Scholar

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64591">Dendrochronological Dating and Provenancing of String Instruments</a>. The Authors section was updated.

Erratum: Dendrochronological Dating and Provenancing of String Instruments

The goal of this study is to present a protocol for the dendrochronological analysis of tree rings on the top plate of a wooden musical stringed instrument. Dendrochronology is used as a method to determine the age of the wood of the instrument by determining the year in which the youngest tree ring on the plate was formed and after which the instrument was made (or before which the instrument could not have been made).
Dating a musical instrument (such as a violin) is an important step in its authentication1,2,3,4,5,6. It is a complex process that involves the year in which the instrument was made, as well as the maker or instrument-making school or geographical area. To do this, dendrochronology is often combined with other techniques, which include the study of the label on the instrument (which is often not reliable) and the inspection of the instrument and its parts such as the outlines, scroll, wood figure and aging, varnish, f-holes, and purfling (Figure 1). The authentication can only be done by experts5,6,7.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64591/64591fig01.jpg" /> 
Figure 1: The top of the violin and its parts. The wood of the top plate (also called the front plate, belly, or soundboard) made of Norway spruce (Picea abies) can be dated by dendrochronology. The characteristics and dimensions of other parts such as the scroll, f-holes, and purfling are studied by organologists and help to authenticate the instrument. Scale = 20 cm. <a href="https://www.jove.com/files/ftp_upload/64591/64591fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
Dendrochronology is the science of dating tree rings in the wood, also called annual rings, growth rings, or growth layers, which are formed every year in the trees of temperate zones. Dendrochronology clarifies in which calendar year a particular tree ring was formed. By dating the outermost and most recently formed tree ring just below the bark, the last year of a tree&#39;s life before it was cut down can be determined.
Dendrochronology is based on the principle that annual variations in tree ring width (and other characteristics) are largely influenced by the environment, especially the climate, in which the trees grow. When conditions are similar over an area, trees of the same species show similar tree ring variations from year to year8. This means that the tree ring series (i.e., the temporal sequence of tree ring widths over time) is similar for trees of the same species in the same area.
The dating of wooden instruments follows the principles used for dating historical objects. In most cases, it is based on measuring tree ring width, creating tree ring series of the same object, cross-dating (to determine their matching position), and averaging them into a floating chronology of an object showing the tree ring series in relative time3,4,6.
Absolute dating (determining the calendar year of tree ring formation) is accomplished by cross-dating with one or more reference chronologies established for a particular tree species and geographic area4,6.&#160;The reference chronologies must be based on the tree ring widths of a sufficient number of trees (replication) and should be long enough to cover the period of interest.
Dendrochronology is regularly applied to determine the age of stringed instruments such as violins, violas, and cellos1,9,10,11,12,13. For stringed instruments, the wood of the top plates (also the front plate, belly, or soundboard) can be dated. They are usually made of Norway spruce (Picea abies) or silver fir (Abies alba)4,6,13. The measurements must be made in a non-invasive way directly on the instrument or by using images. The measurements are usually made at different locations on the top plate to establish a sequence for the instrument that can be dated with reference chronologies.
Dating is the most critical step because a reference chronology must be available for the species, geographic area, and time period of the instrument being studied. Many chronologies are available at the International Tree Ring Data Bank (ITRDB)14,&#160;but only a few are of Norway spruce or silver fir from the region covering the period of interest6; therefore, dendrochronology laboratories put a lot of effort into constructing reference chronologies. The likelihood of dating increases if a network of chronologies is available, including those from exactly defined forest sites, dated instruments, and instrument collections from various manufacturers like the Stradivari, Guarneri, and Amati families from Italy5,6,15,16, Jacob Stainer from Austria, as well as Joachim Tielke and members of the Hoffmann family from Germany17,18,19. The fine historical instruments made by the manufacturers in the 16th century to 18th century are most prized by musicians and collectors, although the importance of many less-known makers is also growing3,4,6,12.
Dendrochronology provides the end date, which must be considered the terminus post quem &#8211;&#160;the year after which the instrument was made. Dendrochronology is also used for dendroprovenancing, which helps to determine the geographic origin of the wood and to assign instruments to specific violin makers or violin-making schools3,4,6.
The dendrochronological end date almost never coincides exactly with the year the instrument was made, and the latter must, thus, be estimated, which requires a lot of supporting information and cooperation between experts of different fields.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CDendro</td>
			<td>Cybis Elektronik &#38; Data AB</td>
			<td>https://www.cybis.se</td>
			<td>program CoDendro for dendro data management and crossdating</td>
		</tr>
		<tr>
			<td>CooRecorder</td>
			<td>Cybis Elektronik &#38; Data AB</td>
			<td>https://www.cybis.se</td>
			<td>program CooRecorder to measure tree ring widths on images</td>
		</tr>
		<tr>
			<td>TSAP-Win</td>
			<td>RINNTECH</td>
			<td>https://rinntech.info/products/tsap-win/</td>
			<td>Time series analysis software</td>
		</tr>
	</tbody>
</table>

dendrochronological dating and provenancing of string instruments

Dendrochronology, the science of dating tree rings in the wood, defines in which calendar year a particular tree ring was formed. The method can be used to determine the age and authentication of wooden musical instruments. We present a protocol describing how to perform a dendrochronological analysis on stringed instruments and how to interpret the dating. The protocol describes the basic steps in the analysis of top plates, which are usually made of Norway spruce (Picea abies) or, more rarely, silver fir (Abies alba). First, the top plate is carefully inspected, and then the tree ring widths are measured directly on the instrument using high-resolution images. After completing the measurements, a tree ring sequence of the instrument is created, and, in the next step, dating is performed with a number of reference chronologies of the tree species from different geographical areas and instruments. The specialists who date the instruments also invest work in creating reference chronologies. The dendrochronological report provides the dating of an instrument as a calendar year (end date), indicating the year in which the last (most recent) tree ring on the top plate was formed when the tree was still alive. The end date represents the terminus post quem, the year after which the instrument was made or before which it could not have been made. To estimate the year of manufacture, one must consider the time required for wood drying and storing and the number of tree rings removed during wood processing. This protocol is intended to help those commissioning such an analysis to better understand how the analysis is performed and how to interpret the dendrochronological reports in terms of the age, origin, maker, and authenticity of the instrument.

A typical case in which a dendrochronological study was requested is a violin allegedly made by Andrea Guarneri of Cremona belonging to the family/school that produced numerous valuable instruments16,24. The violin in question contained two labels. One stated that the instrument was made by Andrea Guarneri of Cremona in 1747, while the other stated only 1867. However, the organological examination of the violin (Figure 3) suggested t...

Watch this Scientific Journal Video about Dendrochronological Dating and Provenancing of String Instruments at JoVE.com

Dendrochronological Dating and Provenancing of String Instruments

The dendrochronological analysis of a stringed instrument requires inspecting the top plate, measuring the tree ring widths, establishing the chronology of the instrument, and dating through determining the end date-the year of the formation of the most recent tree ring.

Dendrochronology, the science of dating tree rings in the wood, defines in which calendar year a particular tree ring was formed. The method can be used ...

Dendrochronology is the science of tree rings in the wood. It can be used to determine the age and origin of wooden musical instruments, like a violin. Dendrochronology clarifies in which calendar year a particular tree ring was formed.We can estimate when the tree was felled, where it was growing, and when the violin was made. This protocol will help you to better understand how the dendrochronological analysis is performed, how to interpret the report, and how to determine whether, or not the violin is original. Dr.Angela Balzano, Nina Skrk Dolar, and Luka Krze, researchers from our laboratory, prepared the demonstration of the procedure.We are collaborating on this project with Blaz Demsar, a violin maker from Ljubljana, Slovenia, who has provided the violins for the demonstration of the protocol. Dr.Micha Beuting, a publicly accredited and sworn expert for dendrochronological investigations on musical instruments by the Chamber of Commerce in Hamburg, Germany, who has helped us with reference chronologies, and demonstrated how a report should look like. To begin, examine the instrument and all its parts.Take detailed photos of the top, back, side, scroll, and label, along with the measuring scale for reporting and archive. Examine the top plate of the instrument to understand its construction. Check whether the top plate is made of one, two, or more parts, and how they are joined together.On the radial board of the top plate, examine the structure and orientation of the tree rings. The rings are seen as bands of annual growth layers, consisting of light colored early wood and darker late wood, and delimited by the tree ring boundaries. Based on the location and transition of early wood to late wood, determine which side is the bark and which side is the pith.Usually, the most recently formed tree rings, which were closer to the bark, are located at the joint in the center of the top plate. First, inspect the widest parts on each piece of the top plate. Check for any damage, repairs, retouching, or dirt, and select the areas where the varnish is transparent enough to see the tree rings.If the instrument is opened and the top plate can be removed, measure the tree rings on the underside of the top plate, where no varnish is applied. Ensure that the selected area has the greatest number of tree rings, and that the boundaries between the rings can be recognized. Using a magnifying lens, or stereo microscope, try to clearly see the tree rings that were near the bark for an accurate identification.Mark the measuring line in the direction from the bark to the pith, and place a measuring scale on the board. For image analysis, place the top plate of the violin on the scanner, and scan the parts selected for the tree ring measurement using a resolution of 1, 200 DPI, or greater, and a high depth of field. For editing the images, use a graphics editing program.Check the quality of the image, and adjust the color, brightness, and contrast to best see the annual growth rings and the boundaries between them, and save the images. Start an image analysis system, preferably one designed for the automatic and manual detection of tree rings and the measurement of tree ring width. Open the image and set the calibration by measuring the distance of a known length on the measurement scale as part of the image.Start the measurement by clicking on the tree ring boundaries, and note whether the first tree ring measured is closer to the bark, or closer to the pith. To measure the distance between the tree rings, click the tree ring boundaries manually if the automatic detection of tree ring width is not possible, or requires too many corrections. Measure all the tree ring widths along the measurement line.Save the data as the tree ring series with each tree ring width recorded for a specific relative year. The output of this measurement is a table of tree ring widths versus time in relative years. For data processing, open the data file containing the tree ring widths in millimeters against years.For cross-dating, open the tree ring series to be cross-dated, and another tree ring series of the same instrument to serve as a reference. Run the cross-dating to move them into the synchronous position. Check for the main cross-dating parameters used in the study of violins.Consider the parameters significant. The Baillie and Pilcher T-value, or TVBP, is a minimum of four, and the Hollstein T-value, or TVH, is at least four, and if the concordance coefficient, or Glk, percent is at least 65%Consider the overlap, which indicates the overlapping period of two tree ring series in years. When the two tree ring series are on the same relative timescale, save the position of the series.Repeat this step until all the tree ring series of each board and instrument are cross-dated. Then to create the chronology of the violin, observe the graphs, and select those that have no measurement arrows. If the agreement between the series within the instrument is statistically significant, average all the tree ring series into one chronology of the violin.Cross-date the chronology of the violin with several reference chronologies. Perform the cross-dating with the reference chronologies. Then check the parameters of the agreement and visually assess the proposed dating proposition.For successful dating, ensure that the same end date is confirmed by multiple chronologies with statistically significant parameters, as well as by visual comparison of the sequences. As the final result of the dendrochronological analysis, report the end date indicating the year in which the last, or most recent tree ring on the top plate was formed when the tree was still alive. Use the information on the original label and other sources to estimate the number of years required for wood drying and storing, and the number of tree rings removed during wood processing.Add this value, which usually ranges between 50 to 20 years, to the end date to propose the potential date of manufacture of the instrument. To validate the authenticity of the violin, confirm the assumptions about the age, maker, and area of origin. Write a report that includes the dendrochronological end date and sufficient information about the investigation that may help in the interpretation of the dating.An historical violin was inscribed with two labels in the inside of the violin body, one stating that the instrument was made by Andrea Guarneri of Cremona in 1747, and the other stating only 1867. However, organological examination of the top, back, and the scroll of the violin suggested that it was probably about 300 years old. The top plate was made of two radial boards of Norway spruce.The tree rings were very narrow, averaging 0.69 millimeters, and the end date of the youngest tree ring on the plate was determined to be 1640. Dendrochronological cross-dating performed using over 110 reference chronologies of spruce covering the periods from 1137 to 2009 revealed an end date of 1640. Tree ring series of the violin showed statistically significant similarity, and the end date 1640 was confirmed with more than 70 reference chronologies from different forest sites and historic buildings, mainly in Austria and Germany, as well as individual instruments and instrument collections of violin makers, like Jacob Stainer.This result presents the dating with a widely used reference chronology of a high elevation alpine stand in Austria, constructed and published by Veronica Siebenlist Kerner in 1984. Thus, the instrument was built after 1640, and is much older than proposed by the inscriptions on the labels. In this procedure, one must clearly see the tree ring for correct measurements.Also, adequate reference chronologies are needed for dating. Dendrochronology is often combined with other techniques, like the study of the label, and the inspection of the instrument and its parts. Dendrochronology benefits from the improvement of imaging techniques and promotes the development of networks of reference chronologies, and also the improvement of dendroprovenancing methods.

dendrochronological-dating-and-provenancing-of-string-instruments

Research

JoVE Journal

Environment

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

Characterization and Application of Passive Samplers for Monitoring of Pesticides in Water

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study provides a protocol for the passive sampler preparation, calibration, extraction method and instrumental analysis. Sampling rates (RS) and passive sampler-water partition coefficients (KPW) were calculated for silicone rubber, polar organic chemical integrative sampler POCIS-A, POCIS-B, SDB-RPS and C18 disk. The uptake of the selected compounds depended on their physicochemical properties, i.e., silicone rubber showed a better uptake for more hydrophobic compounds (log octanol-water partition coefficient (KOW) &#62; 5.3), whereas POCIS-A, POCIS-B and SDB-RPS disk were more suitable for hydrophilic compounds (log KOW &#60; 0.70).

A protocol about the characterization and application of five different passive sampling devices is presented.

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study ...

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 ...

Extraction and Characterization of Surfactants from Atmospheric Aerosols

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in the Earth&#39;s atmosphere, a central process in meteorology, hydrology, and for the climate system. But because specific extraction and characterization of these compounds have been lacking for decades, very little is known on their identity, properties, mode of action and origins, thus preventing the full understanding of cloud formation and its potential links with the Earth&#39;s ecosystems.
In this paper we present recently developed methods for 1) the targeted extraction of all the surfactants from atmospheric aerosol samples and for the determination of 2) their absolute concentrations in the aerosol phase and 3) their static surface tension curves in water, including their Critical Micelle Concentration (CMC). These methods have been validated with 9 references surfactants, including anionic, cationic and non-ionic ones. Examples of results are presented for surfactants found in fine aerosol particles (diameter &#60;1 &#956;m) collected at a coastal site in Croatia and suggestions for future improvements and other characterizations than those presented are discussed.

Methods are presented for the targeted extraction of surfactants present in atmospheric aerosols and the determination of their absolute concentrations and surface tension curves in water, including their Critical Micelle Concentration (CMC).

Surface-active compounds, or surfactants, present in atmospheric aerosols are expected to play important roles in the formation of liquid water clouds in ...

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic pathway of dimethylarsinic acid (DMAV) thiolation, have been recently found in the environment as well as human organs. DMMTAV and DMDTAV can be quantified to determine the ecological effects of dimethylated thioarsenicals and their stability in environmental media. The synthesis method for these compounds is unstandardized, making replicating previous studies challenging. Furthermore, there is a lack of information about storage techniques, including storage of compounds without species transformation. Moreover, because only limited information about synthesis methods is available, there may be experimental difficulties in synthesizing standard chemicals and performing quantitative analysis. The protocol presented herein provides a practically modified synthesis method for the dimethylated thioarsenicals, DMMTAV and DMDTAV, and will help in the quantification of species separation analysis using high performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The experimental steps of this procedure were modified by focusing on the preparation of chemical reagents, filtration methods, and storage.

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

This article presents modified experimental protocols for dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV) synthesis, inducing dimethylarsinic acid (DMAV) thiolation through mixing of DMAV, Na2S, and H2SO4. The modified protocol provides an experimental guideline, thereby overcoming limitations of the synthesis steps that could have caused experimental failures in quantitative analysis.

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical data that may be used in predictive modelling to forecast evolutionary responses of natural ecosystems to future environmental changes. However, excluding a few exceptional cases, long-term studies are scarce because of logistic difficulties associated with accessing temporal samples. Temporal dynamics are frequently studied in the laboratory or in controlled mesocosm experiments with exceptional studies that reconstruct the evolution of natural populations in the wild.
Here, a standard operating procedure (SOP) is provided to revive or resurrect dormant Daphnia magna, a widespread zooplankton keystone species in aquatic ecosystems, to dramatically advance the state-of-the-art longitudinal data collection in natural systems. The field of Resurrection Ecology was defined in 1999 by Kerfoot and co-workers, even though the first attempts at hatching diapausing zooplankton eggs date back to the late 1980s. Since Kerfoot&#39;s seminal paper, the methodology of resurrecting zooplankton species has been increasingly frequently applied, though propagated among laboratories only via direct knowledge transfer. Here, an SOP is described that provides a step-by-step protocol on the practice of resurrecting dormant Daphnia magna eggs.
Two key studies are provided in which the fitness response of resurrected Daphnia magna populations to warming is measured, capitalizing on the ability to study historical and modern populations in the same settings. Finally, the application of next generation sequencing technologies to revived or still dormant stages is discussed. These technologies provide unprecedented power in dissecting the processes and mechanisms of evolution if applied to populations that have experienced changes in selection pressure over time.

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical ...

Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several limitations that have resulted in the exclusion of many species. Advances in the field over the last two decades have led occupational health researchers to turn to molecular-based approaches for identifying fungal hazards. These methods have resulted in the detection of many species within indoor and occupational environments that have not been detected using traditional methods. This protocol details an approach for determining fungal diversity within air samples through genomic DNA extraction, amplification, sequencing, and taxonomic identification of fungal internal transcribed spacer (ITS) regions. ITS sequencing results in the detection of many fungal species that are either not detected or difficult to identify to species level using culture or microscopy. While these methods do not provide quantitative measures of fungal burden, they offer a new approach to hazard identification and can be used to determine overall species richness and diversity within an occupational environment.

Determining the fungal diversity within an environment is a method utilized in occupational health studies to identify health hazards. This protocol describes DNA extraction from occupational air samples for amplification and sequencing of fungal ITS regions. This approach detects many fungal species that can be overlooked by traditional assessment methods.

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several ...

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those of suspended/sinking particles. Here, we present an all plastic (Polypropylene and Polycarbonate), multi-layer filtration system for collection of suspended particulate matter (SPM) at sea. This ultra-clean sampling device has been designed and developed specifically for trace element studies. Meticulous selection of all non-metallic materials and utilization of an in-line flow-through procedure minimizes any possible metal contamination during sampling. This system has been successfully tested and tweaked for determining trace metals (e.g., Fe, Al, Mn, Cd, Cu, Ni) on particles of varying size in coastal and open ocean waters. Results from the South China Sea at the South East Asia Time-Series (SEATS) station indicate that diurnal variations and spatial distribution of plankton in the euphotic zone can be easily resolved and recognized. Chemical analysis of size-fractionated particles in surface waters of the Taiwan Strait suggests that the larger particles (&#62;153 &#181;m) were mostly biologically derived, while the smaller particles (10 - 63 &#181;m) were mostly composed of inorganic matter. Apart from Cd, the concentrations of metals (Fe, Al, Mn, Cu, Ni) decreased with increasing size.

Plankton and suspended particles play a major role in the biogeochemical cycles in the ocean. Here, we provide an ultra-clean, low stress method for the collection of various sizes of particles and plankton at sea with the capability of handling large volumes of seawater.

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those ...

Quantification of Endogenous Auxin and Cytokinin During Internode Culture of Ipecac

Adventitious shoot formation is an important technique for the propagation of economically important crops and for the regeneration of transgenic plants. Phytohormone treatment is required for the induction of adventitious shoots in most species. Whether adventitious shoots can be induced is determined by the balance between auxin and cytokinin (CK) levels. Much effort goes into determining optimum concentrations and combinations of phytohormones in each tissue used as explants and in each plant species. In ipecac, however, adventitious shoots can be induced on internodal segments in culture medium without phytohormone treatment. This allows the inherent plasticity of ipecac for cell differentiation to be evaluated. To induce adventitious shoots in ipecac, we cultured internodal segments at 24 &#176;C under 15 &#181;mol m&#8722;2 s&#8722;1 of light in a 14-h light/10-h dark cycle on phytohormone-free B5 medium solidified with 0.2% gellan gum for 5 weeks. To investigate phytohormone dynamics during adventitious shoot formation, we measured endogenous indole-3-acetic acid and CKs in the segments by liquid chromatography-tandem mass spectrometry LC-MS/MS. This method allows analysis of endogenous indole-3-acetic acid and CKs levels in a simple manner. It can be applied to investigate the dynamics of endogenous auxin and CK during organogenesis in other plant species.

Adventitious shoots can be induced on internodal segments of ipecac without phytohormone treatment. To evaluate phytohormone dynamics during adventitious shoot formation, we measured endogenous auxin and cytokinin in internodal segments by LC-MS/MS.

Adventitious shoot formation is an important technique for the propagation of economically important crops and for the regeneration of transgenic plants. ...

Laser-Induced Fluorescence Emission (L.I.F.E.) as Novel Non-Invasive Tool for In-Situ Measurements of Biomarkers in Cryospheric Habitats

Global warming affects microbial communities in a variety of ecosystems, especially cryospheric habitats. However, little is known about microbial-mediated carbon fluxes in extreme environments. Hence, the methodology of sample acquisition described in the very few studies available implies two major problems: A) high resolution data require a large number of samples, which is difficult to obtain in remote areas; B) unavoidable sample manipulation such as cutting, sawing, and melting of ice cores that leads to a misunderstanding of in situ conditions. In this study, a prototype device that requires neither sample preparation nor sample destruction is presented. The device can be used for in situ measurements with a high spectral and spatial resolution in terrestrial and ice ecosystems and is based on the Laser-Induced Fluorescence Emission (L.I.F.E.) technique. Photoautotrophic supraglacial communities can be identified by the detection of L.I.F.E. signatures in photopigments. The L.I.F.E. instrument calibration for the porphyrin derivates chlorophylla (chla) (405 nm laser excitation) and B-phycoerythrin (B-PE) (532 nm laser excitation) is demonstrated. For the validation of this methodology, L.I.F.E. data were ratified by a conventional method for chla quantification that involved pigment extraction and subsequent absorption spectroscopy. The prototype applicability in the field was proven in extreme polar environments. Further testing on terrestrial habitats took place during Mars analog simulations in the Moroccan dessert and on an Austrian rock glacier. The L.I.F.E. instrument enables high resolution scans of large areas with acceptable operation logistics and contributes to a better understanding of the ecological potential of supraglacial communities in the context of global change.

Laser-Induced Fluorescence Emission L.I.F.E. as Novel Non-Invasive Tool for In-Situ Measurements of Biomarkers in Cryospheric Habitats

Carbon fluxes in the cryosphere are hardly assessed yet but are crucial regarding climate change. Here we show a novel prototype device that captures the phototrophic potential in supraglacial environments based on laser-induced fluorescence emission (L.I.F.E.) technology offering high spectral and spatial resolution data under in situ conditions.