Un merci spécial à la Division St. Regis Mohawk Environmental, Welaka National Fish Hatchery, US Fish and Wildlife Service Research Unit, Southern Illinois University, US Fish and Northeast Fisheries Center de la faune, Sterling Caviar, Université de Californie-Davis, Garrison Dam National Fish écloserie, Bears Bluff national Fish Hatchery, et US Fish and Wildlife service Panama office Resource Ville des pêches pour leur aide à fournir l&#39; esturgeon pour ce projet. cet article est Contribution 2013 du Centre des sciences de l&#39; USGS des Grands Lacs.

Remarque importante: En raison de l&#39;état de protection de toutes les espèces d&#39;esturgeons aux États-Unis, permet d&#39;acquérir et de gérer les poissons ont été obtenus avant la procédure de laboratoire. Tous les spécimens ont été acquis à partir de sources d&#39;écloserie post mortem dues à des causes naturelles et éliminés en suivant la procédure standard. 1. Préparation des échantillons <ol><li> Avant toute incisions pondent carcasse de l&#39; esturgeon sur le côté et à l&#39; aide d&#39; un ruban à mesurer pour mesurer la longueur totale du museau à l&#39;extrémité de la queue à la 1 mm le plus proche. (Figure 1A) </li><li> Peser la carcasse de l&#39;esturgeon sur une échelle taré à 1 g près. Évaluer la carcasse pour toutes les balises ou des marques extérieures et enregistrement. </li></ol> 2. Otolith Extraction <ol><li> Faire une incision verticale avec un grand couteau de filet entre la base du crâne et la première scute dorsale à travers le corps pour séparer la tête du corps (figure 1B). Après avoir enlevé la tête, turn la tête de sorte que la bouche est orientée vers le haut (figure 1C). </li><li> Retirer les tissus mous de la bouche et les branchies en utilisant le couteau filet pour couper le long de la face ventrale du crâne (figure 1D). Faire une incision avec une scie à os électrique à partir de la pointe du museau à la base du crâne à bisect complètement la tête (figure 1E). Séparer les deux moitiés manuellement afin d&#39; exposer la cavité du cerveau (figure 2A). </li><li> Retirez et la matière cérébrale (encadré rouge à gauche) avec une pince pour voir canaux semi - circulaires (boîte rouge à droite) dans la cavité du cerveau (figure 2B) jeter. </li><li> Enlever le cartilage du canal semi - circulaire de la cavité du cerveau avec une pince (figure 2C). Percer soigneusement cartilage canal semi - circulaire avec la pointe de la pince et enlever otolithes (figure 2D). Retirer les otolithes (structure osseuse dur à l&#39; intérieur du cartilage) avec une pince du cartilage canal semi - circulaire (figure 2E) </strong&gt;. </li><li> Répétez les étapes 02.03 à 02.04 pour enlever otolithes de l&#39;autre moitié du crâne. </li><li> Tremper les otolithes dans 92% d&#39;éthanol pendant 5 secondes pour éliminer tout cartilage. La place otolithes dans un 25 ml flacon à scintillation ouvert à sécher pendant 24 heures. </li></ol>

<ol>
	<li>Auer, N. A., LeBreton, G. T. O., Beamish, F. W. H., McKinley, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conservation.">Conservation.</a> Sturgeons and Paddlefish of North America. 27, 252-276 (2004).</li><li>Campana, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy,+precision+and+quality+control+in+age+determination,+including+a+review+of+the+use+and+abuse+of+age+validation+methods).">Accuracy, precision and quality control in age determination, including a review of the use and abuse of age validation methods).</a> Journal of Fish Biology. 59, 197-242 (2001).</li><li>Bruch, R. M., Campana, S. E., Davis-Foust, S. L., Hansen, M. J., Janssen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lake+Sturgeon+age+validation+using+bomb+radiocarbon+and+known-age+fish.">Lake Sturgeon age validation using bomb radiocarbon and known-age fish.</a> Transactions of the American Fisheries Society. 138, 361-372 (2009).</li><li>Baremore, I. E., Rosati, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+validated,+minimally+deleterious+method+for+ageing+sturgeon.">A validated, minimally deleterious method for ageing sturgeon.</a> Fishery Bulletin. 112, 274-282 (2014).</li><li>Stevensen, J. T., Secor, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age+determination+and+growth+of+Hudson+River+Atlantic+sturgeon,+Acipenser+oxyrhynchus.">Age determination and growth of Hudson River Atlantic sturgeon, Acipenser oxyrhynchus.</a> Fishery Bulletin. 97, 153-166 (1999).</li><li>Brennan, J. S., Cailliet, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+age-determination+techniques+for+white+sturgeon+in+California.">Comparative age-determination techniques for white sturgeon in California.</a> Transactions of the American Fishery Society. 118, 296-310 (1989).</li><li>Rossiter, A., Noakes, D. L. G., Beamish, F. W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+age+estimation+for+the+lake+sturgeon.">Validation of age estimation for the lake sturgeon.</a> Transactions of the American Fisheries Society. 124, 777-781 (1995).</li><li>Chalupnicki, M. A., Dittman, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=North+American+Sturgeon+Otolith+Morphology.">North American Sturgeon Otolith Morphology.</a> Copeia. , 260-266 (2016).</li><li>Secor, D. H., Dean, J. M., Laban, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Manual for otolith removal and preparation for microstructural examination. , (1991).</li></ol>

Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the U.S. Government.

La vue d&#39;ensemble d&#39;une méthode alternative d&#39;extraction otolithes des carcasses d&#39;esturgeons a été détaillée. Il est important de noter qu&#39;une attention particulière est nécessaire pour la mise en place d&#39;incision latérale sur la ligne médiane du crâne afin de bisect le crâne uniformément pour assurer aucun mal aux otolithes lors de la dissection. Si l&#39;incision est pas assez profonde pour bisect complètement le crâne, il sera très difficile d&#39;exposer la cavité du cerveau ...

Sturgeon (Acipenseridae) populations à travers le monde sont en baisse depuis plus d&#39; un siècle en raison des impacts , y compris la perte d&#39;habitat, la fragmentation de la population, et la surpêche telle que de nombreuses populations sont protégés par les lois fédérales et d&#39;état 1,2. Les organismes de gestion à travers les Etats - Unis et le monde ont identifié l&#39; esturgeon comme une espèce cible pour la restauration et la récupération 1. Caractéristiques de la population (croissance, recrutement, régime alimentaire) de la plupart des populations d&#39;esturgeons ont été étudiés afin de mieux comprendre la biologie et l&#39; histoire de vie caractéristiques de base 1. En raison du statut protégé de la plupart des populations d&#39;esturgeons, l&#39;évaluation de l&#39;âge est difficile dans les petites populations menacées où aucun sacrifice ou de nuire à des individus est justifiée. L&#39;extraction des structures osseuses dans le but d&#39;estimation de l&#39; âge et de l&#39; analyse de la croissance a été un .Utilisation de sections de rayons ailettes technique courante 3 pour déterminer l&#39; âge de Sturgesur est devenu une méthode non - létaux pratique, mais la vérification de la technique a été limitée et l&#39; âge peuvent être très difficiles à déchiffrer chez les personnes âgées (&gt; 15 ans) 4. Les chercheurs évaluant l&#39;utilisation de plusieurs structures osseuses de l&#39;esturgeon pour leur utilisation dans la validation de l&#39;âge de Gulf Sturgeon (Acipenser oxyrinchus desoti) 5, l&#39; esturgeon noir (A. o. Oxyrinchus) 6, l&#39; esturgeon blanc (A. transmontanus) 7, et le lac Sturgeon (A. fluvescens) 8 a conclu que le premier pectoral marginal ray fin fourni l&#39; âge le plus précis , mais a conclu cette procédure invasive pour être néfaste pour les individus plus grands. Actuellement, il est le développement de l&#39; utilisation de mesures de forme otolithes pour distinguer entre les espèces nord - américaines de l&#39; esturgeon avec un taux très élevé de succès 9. Les détails sur le processus d&#39;élimination des otolithes avec une incision transversale dorsale à la base du crâne est un otolithes commun supplémentaireméthode de ction trouvé dans la littérature et clairement détaillé par Secor et al. (1991) 10. Cependant, cette méthode est difficile à mettre en œuvre sur l&#39;esturgeon qui ont très épais avec des crânes otolithes qui sont petits et délicats et auraient tendance à se briser. Un nouvel examen de la littérature montre aucune autre méthodologie détaillée du processus d&#39;élimination des otolithes pour l&#39;esturgeon. L&#39;objectif de cet article était de détail une méthode alternative d&#39;extraction otolithes de l&#39;esturgeon en entrant dans la cavité du cerveau de la face ventrale.

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Measuring Board</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Metric meter stick is preferred</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Weighing scale</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Filleting Knife</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Forceps</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sample vials</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>25 ml scintillation vials work well</td>
		</tr>
	</tbody>
</table>

alternative method of removing otoliths from sturgeon

Rapatrier les otolithes (os de l&#39;oreille) de poissons qui ont des crânes très épais peuvent être temps difficile et très coûteuse. La pratique courante de faire une incision verticale transversale sur le sommet du crâne avec une main ou une scie électrique peut endommager le otolithes si pas effectué correctement. Esturgeons (Acipenseridae) sont une famille en particulier qui ont un crâne très grand et épais. Une nouvelle méthode de laboratoire entrant dans la cavité du cerveau de la face ventrale du poisson pour exposer les otolithes était plus facile que d&#39;autres méthodes d&#39;extraction des otolithes trouvés dans la littérature. Méthodes examinées dans la littérature sont conçus pour le terrain et sont plus efficaces pour le traitement de grandes quantités de poissons rapidement. Cependant, cette nouvelle technique a été conçu pour être plus adapté à un environnement de laboratoire lorsque le temps est pas urgent et extraction réussie de chaque échantillon est critique. Le succès de trouver et supprimer les otolithes en utilisant cette technique est très élevé et ne compromet pas les structure d&#39;une manière quelconque. Cette technique alternative est applicable à d&#39;autres espèces de poissons similaires pour extraire les otolithes.

La taille adulte de l&#39;esturgeon dans le monde varie considérablement, mais l&#39;emplacement des otolithes est cohérente. L&#39;utilisation d&#39;un couteau filet pointu a été trouvé pour éliminer les tissus mous de la bouche facilement illustré à la figure 1D. Une scie à os électrique est l&#39;outil privilégié pour l&#39;incision latérale pour exposer la cavité du cerveau dans la figure 1E. Cependant, l&#39;attention à la bissectrice...

Watch this Scientific Journal Video about Alternative Method of Removing Otoliths from Sturgeon at JoVE.com

Alternative Method of Removing Otoliths from Sturgeon

The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses.

Méthode alternative de Retrait otolithes de Sturgeon

Extracting the otoliths (ear bones) from fish that have very thick skulls can be difficult and very time consuming. The common practice of making a ...

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Environment

8.4K vues.  Great Lakes Science Center. The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses. 

Vidéo: Alternative Method of Removing Otoliths from Sturgeon

Detection of Foodborne Bacterial Pathogens from Individual Filth Flies

There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a particular foodborne illness outbreak has been challenging. This is because insects are not being aseptically collected as part of an environmental sampling program during foodborne outbreak investigations and because there is not a standardized method to detect foodborne bacteria from individual insects. To take a step towards solving this problem, we adapted a protocol from a commercially available PCR-based system that detects foodborne pathogens from food and environmental samples, to detect foodborne pathogens from individual flies.Using this standardized protocol, we surveyed 100 wild-caught flies for the presence of Cronobacter spp., Salmonella enterica, and Listeria monocytogenes and demonstrated that it was possible to detect and further isolate these pathogens from the body surface and the alimentary canal of a single fly. Twenty-two percent of the alimentary canals and 8% of the body surfaces from collected wild flies were positive for at least one of the three foodborne pathogens. The prevalence of Cronobacter spp. on either body part of the flies was statistically higher (19%) than the prevalence of S. enterica (7%) and L.monocytogenes (4%). No false positives were observed when detecting S. enterica and L. monocytogenes using this PCR-based system because pure bacterial cultures were obtained from all PCR-positive results. However, pure Cronobacter colonies were not obtained from about 50% of PCR-positive samples, suggesting that the PCR-based detection system for this pathogen cross-reacts with other Enterobacteriaceae present among the highly complex microbiota carried by wild flies. The standardized protocol presented here will allow laboratories to detect bacterial foodborne pathogens from aseptically collected insects, thereby giving public health officials another line of evidence to find out how the food was contaminated when performing foodborne outbreak investigations.

A PCR-based protocol was adapted to detect Cronobacter spp., Salmonella enterica, and Listeria monocytogenes from body surfaces and alimentary canals of individual wild-caught flies. The goal of this protocol is to detect and isolate bacterial pathogens from individual insects collected as part of an environmental sampling program during foodborne outbreak investigations.

Détection de bactéries pathogènes d&#39;origine alimentaire de mouches individuelle Filth

There is unanimous consensus that insects are important vectors of foodborne pathogens. However, linking insects as vectors of the pathogen causing a ...

Recherche

Environnement

Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may be treated with simple systems designed to promote sorption of contaminants to vegetation and soils and promote infiltration. Two example systems are described: a bioswale treatment system for urban stormwater treatment, and a vegetated drainage ditch for treating agriculture irrigation runoff. Both have similar attributes that reduce contaminant loading in runoff: vegetation that results in sorption of the contaminants to the soil and plant surfaces, and water infiltration. These systems may also include the integration of granulated activated carbon as a polishing step to remove residual contaminants. Implementation of these systems in agriculture and urban watersheds requires system monitoring to verify treatment efficacy. This includes chemical monitoring for specific contaminants responsible for toxicity. The current paper emphasizes monitoring of current use pesticides since these are responsible for surface water toxicity to aquatic invertebrates.

This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

Systèmes de traitement végétal pour éliminer les contaminants associés à la toxicité des eaux de surface dans l&#39;agriculture et le ruissellement urbain

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

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).

Extraction et caractérisation des tensio-actifs de l&#39;atmosphère Aérosols

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

Metal Corrosion and the Efficiency of Corrosion Inhibitors in Less Conductive Media

Material corrosion can be a limiting factor for different materials in many applications. Thus, it is necessary to better understand corrosion processes, prevent them and minimize the damages associated with them. One of the most important characteristics of corrosion processes is the corrosion rate. The measurement of corrosion rates is often very difficult or even impossible especially in less conductive, non-aqueous environments such as biofuels. Here, we present five different methods for the determination of corrosion rates and the efficiency of anti-corrosion protection in biofuels: (i) a static test, (ii) a dynamic test, (iii) a static test with a reflux cooler and electrochemical measurements (iv) in a two-electrode arrangement and (v) in a three-electrode arrangement. The static test is advantageous due to its low demands on material and instrumental equipment. The dynamic test allows for the testing of corrosion rates of metallic materials at more severe conditions. The static test with a reflux cooler allows for the testing in environments with higher viscosity (e.g., engine oils) at higher temperatures in the presence of oxidation or an inert atmosphere. The electrochemical measurements provide a more comprehensive view on corrosion processes. The presented cell geometries and arrangements (the two-electrode and three-electrode systems) make it possible to perform measurements in biofuel environments without base electrolytes that could have a negative impact on the results and load them with measurement errors. The presented methods make it possible to study the corrosion aggressiveness of an environment, the corrosion resistance of metallic materials, and the efficiency of corrosion inhibitors with representative and reproducible results. The results obtained using these methods can help to understand corrosion processes in more detail to minimize the damages caused by corrosion.

The testing of processes associated with material corrosion can often be difficult especially in non-aqueous environments. Here, we present different methods for short-term and long-term testing of corrosion behavior of non-aqueous environments such as biofuels, especially those containing bioethanol.

La Corrosion des métaux et l’efficacité des inhibiteurs de Corrosion dans les milieux moins conductrice

Material corrosion can be a limiting factor for different materials in many applications. Thus, it is necessary to better understand corrosion processes, ...

Detection of Viruses from Bioaerosols Using Anion Exchange Resin

This protocol demonstrates a customized bioaerosol sampling method for viruses. In this system, anion exchange resin is coupled with liquid impingement-based air sampling devices for efficacious concentration of negatively-charged viruses from bioaerosols. Thus, the resin serves as an additional concentration step in the bioaerosol sampling workflow. Nucleic acid extraction of the viral particles is then performed directly from the anion exchange resin, with the resulting sample suitable for molecular analyses. Further, this protocol describes a custom-built bioaerosol chamber capable of generating virus-laden bioaerosols under a variety of environmental conditions and allowing for continuous monitoring of environmental variables such as temperature, humidity, wind speed, and aerosol mass concentration. The main advantage of using this protocol is increased sensitivity of viral detection, as assessed via direct comparison to an unmodified conventional liquid impinger. Other advantages include the potential to concentrate diverse negatively-charged viruses, the low cost of anion exchange resin (~$0.14 per sample), and ease of use. Disadvantages include the inability of this protocol to assess infectivity of resin-adsorbed viral particles, and potentially the need for the optimization of the liquid sampling buffer used within the impinger.

An anion exchange resin-based method, adapted to liquid impingement-based bioaerosol sampling of viruses is demonstrated. When coupled with downstream molecular detection, the method allows for facile and sensitive detection of viruses from bioaerosols.

Détection des virus de bioaérosols à l’aide de résine échangeuse d’anions

This protocol demonstrates a customized bioaerosol sampling method for viruses. In this system, anion exchange resin is coupled with liquid ...

A Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci.

A Loop-mediated Isothermal Amplification LAMP Assay for Rapid Identification of Bemisia tabaci

This paper reports the protocol for a rapid identification assay for Bemisia tabaci based on loop-mediated isothermal amplification (LAMP) technology. The protocol requires minimal laboratory training and can, therefore, be implemented on-site at points of entry for plant imports such as seaports and airports.

Un test d’Amplification isotherme (lampe) médiée par la boucle pour une Identification rapide de Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many ...

Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of the woodboring beetle, Chlorophorus caragana Xie &#38; Wang (2012), for both types of electron microscopy. The scanning electron microscopy (SEM) sample preparation protocol was based on sample chemical fixation, dehydration in a series of ethanol baths, drying, and sputter-coating. By adding Tween 20 (Polyoxyethylene sorbitan laurate) to the fixative and the wash solution, the insect body surface of woodboring beetle was washed more cleanly in SEM. This study&#39;s transmission electron microscopy (TEM) sample preparation involved a series of steps including fixation, ethanol dehydration, embedding in resin, positioning using fluorescence microscopy, sectioning, and staining. Fixative with Tween 20 enabled penetrate the insect body wall of woodboring beetle more easily than it would had been without Tween 20, and subsequently better fixed tissues and organs in the body, thus yielded clear transmission electron microscope observations of insect sensilla ultrastructures. The next step of this preparation was determining the positions of insect sensilla in the sample embedded in the resin block by using fluorescence microscopy to increase the precision of target sensilla positioning. This improved slicing accuracy.

In order to observe ultrastructure of insect sensilla, scanning and transmission electron microscopy (SEM and TEM, respectively) sample preparation protocol were presented in the study. Tween 20 was added into the fixative to avoid sample deformation in SEM. Fluorescence microscopy was helpful for improving slicing accuracy in TEM.

Méthode de préparation d'échantillons de microscope électronique de balayage et de transmission pour les appendices du bec de bois

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of ...

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) and quantify any differences in fin cortisol levels among three main sturgeon species. Fins were harvested from 19 sacrificed sturgeons including seven beluga (Huso huso), seven Siberian (Acipenser baerii), and five sevruga (A. stellatus). The sturgeons were raised in Iranian farms for 2 years (2017-2018), and cortisol extraction analysis was conducted in South Korea (January-February 2019). Jawbones from five H. huso were also used for cortisol extraction. Data were analyzed using the general linear model (GLM) procedure in the SAS environment. The intra- and inter-assay coefficients of variation were 14.15 and 7.70, respectively. Briefly, the cortisol extraction technique involved washing the samples (300 &#177; 10 mg) with 3 mL of solvent (ultrapure water and isopropanol) twice, rotation at 80&#160;rpm for 2.5 min, air-drying the washed samples at room temperature (22-28 &#176;C) for 7 days, further drying the samples using a bead beater at 50 Hz for 32 min and grinding them into powder, applying 1.5 mL methanol to the dried powder (75 &#177; 5 mg), and slow rotation (40 rpm) for 18 h at room temperature with continuous mixing. Following extraction, samples were centrifuged (9,500 x g for 10 min), and 1 mL supernatant was transferred into a new microcentrifuge tube (1.5 mL), incubated at 38 &#176;C to evaporate the methanol, and analyzed via enzyme-linked immunosorbent assay (ELISA). No differences were observed in fin cortisol levels among species or in fin and jawbone cortisol levels between washing solvents. The results of this study demonstrate that the sturgeon jawbone matrix is a promising alternative stress indicator to solid matrices.

In this study, we present a protocol for cortisol extraction from the fin and jawbone of sturgeon species. Fin and jawbone cortisol levels were further examined by comparing two washing solvents followed by ELISA assays. This study piloted the feasibility of jawbone cortisol as a novel stress indicator.

Extraction de cortisol à partir de matrices de nageoires d'esturgeon et de mâchoires

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) ...

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

Une méthode pour quantifier les arthropodes foliage-dwelling

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

Electrostatic Method to Remove Particulate Organic Matter from Soil

Estimations of soil organic carbon are dependent on soil processing methods including removal of undecomposed plant material. Inadequate separation of roots and plant material from soil can result in highly variable carbon measurements. Methods to remove the plant material are often limited to the largest, most visible plant materials. In this manuscript we describe how electrostatic attraction can be used to remove plant material from a soil sample. An electrostatically charged surface passed close to dry soil naturally attracts both undecomposed and partially decomposed plant particles, along with a small quantity of mineral and aggregated soil. The soil sample is spread in a thin layer on a flat surface or a soil sieve. A plastic or glass Petri dish is electrostatically charged by rubbing with polystyrene foam or nylon or cotton cloth. The charged dish is passed repeatedly over the soil. The dish is then brushed clean and recharged. Re-spreading the soil and repeating the procedure eventually results in a diminishing yield of particulates. The process removes about 1 to 5% of the soil sample, and about 2 to 3 times that proportion in organic carbon. Like other particulate removal methods, the endpoint is arbitrary and not all free particulates are removed. The process takes approximately 5 min and does not require a chemical process as do density flotation methods. Electrostatic attraction consistently removes material with higher than average C concentration and C:N ratio, and much of the material can be visually identified as plant or faunal material under a microscope.

Removing recently deposited and incompletely decomposed plant material from soil samples reduces the influence of temporary seasonal inputs on soil organic carbon measurements. Attraction to an electrostatically charged surface can be used to quickly remove a substantial amount of particulate organic matter.

Procédé électrostatique pour éliminer les matières organiques particulaires du sol

Estimations of soil organic carbon are dependent on soil processing methods including removal of undecomposed plant material. Inadequate separation of ...

Méthode alternative de Retrait otolithes de Sturgeon

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