Un ringraziamento speciale a Divisione St. Regis Mohawk ambientale, Welaka Nazionale vivaio, US Fish and Wildlife Service Unità di Ricerca, Southern Illinois University, US Fish and Wildlife Service nord-est Centro per la pesca, Sterling caviale, University of California-Davis, Garrison Dam Pesce Nazionale Hatchery, Orsi Bluff National Fish Hatchery, e US Fish and Wildlife Service di Panama Office Resource Città della pesca per il loro aiuto la fornitura di storione per questo progetto. in questo articolo è il contributo 2013 del USGS Great Lakes Science center.

Nota importante: A causa della condizione protetta di tutte le specie di storione negli Stati Uniti, permette di acquisire e gestire pesci sono stati ottenuti prima procedura di laboratorio. Tutti i campioni sono stati acquisiti da fonti di incubazione post mortem dovute a cause naturali e smaltiti seguendo la procedura standard. 1. Preparazione del campione <ol><li> Prima eventuali incisioni giacevano storione carcassa su un lato e con un nastro di misurazione per misurare la lunghezza totale del muso alla punta della coda per l&#39;approssimazione di 1 mm. (Figura 1A) </li><li> Pesare storione carcassa su una scala tarato l&#39;approssimazione di 1 g. Valutare carcassa per eventuali variabili esterne o segni e record. </li></ol> 2. Estrazione Otolith <ol><li> Effettuare una incisione verticale con una lama del raccordo tra la base del cranio e il primo scute dorsali attraverso il corpo per separare la testa dal corpo (Figura 1B). Dopo aver rimosso la testa, turn la testa in modo che la bocca è rivolta verso l&#39;alto (Figura 1C). </li><li> Rimuovere tessuti molli della bocca e branchie utilizzando il coltello filetto per tagliare lungo la faccia ventrale del cranio (Figura 1D). Fare un&#39;incisione con una sega elettrica osso dalla punta del muso alla base del cranio alla bisettrice completamente la testa (Figura 1E). Separare le due metà manualmente per esporre la cavità cerebrale (Figura 2A). </li><li> Rimuovere e materia cerebrale (Red box a sinistra) con una pinza per visualizzare canali semicircolari (Red box a destra) all&#39;interno della cavità cerebrale (Figura 2B) scartare. </li><li> Rimuovere semicircolare cartilagine canale dalla cavità cerebrale con una pinza (Figura 2C). Forare con cautela semicircolare cartilagine canale con la punta della pinza e rimuovere otoliti (Figura 2D). Rimuovere il otoliti (struttura ossea duro dentro la cartilagine) con una pinza dalla cartilagine canale semicircolare (Figura 2E) </strong&gt;. </li><li> Ripetere i punti 2.3 - 2.4 a rimuovere otolite dall&#39;altra metà del cranio. </li><li> Immergere le otoliti in 92% di etanolo per 5 secondi per eliminare ogni cartilagine. Luogo otoliti in un 25 ml di scintillazione flacone aperto ad asciugare per 24 ore. </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 panoramica di un metodo alternativo di estrazione di otoliti dalle carcasse di storione è stato dettagliato. È importante notare che una particolare attenzione è necessaria per il posizionamento incisione laterale sulla linea mediana del cranio per bisettrice cranio uniformemente per garantire nessun danno agli otoliti durante la dissezione. Se l&#39;incisione non è abbastanza profonda per bisettrice completamente cranio sarà molto difficile da esporre la cavità cerebrale dove la posizione delle otoliti è chiaramente visibile. Con l&#39;esperienza, la tecnica generale è relativamente semplice e conduce ad una elevata percentuale di otoliti essere trovati intatti. Questo nuovo metodo consente ricerche un modo alternativo per estrarre otoliti per le specie diverse da storione che può essere difficile usando metodi di estrazione otoliti standard. Le limitazioni di questo metodo sono l&#39;applicabilità ad una impostazione non per l&#39;uso sul campo in laboratorio. Il tempo necessario per la preparazione della carcassa non è favorevole a campioni raccoltinel campo in modo tempestivo. Un&#39;altra preoccupazione è l&#39;uso sulle specie di storione che sono in genere protetti ai sensi statali o federali a causa di una diminuzione della popolazione. È molto difficile ottenere permessi di condurre questo tipo di valutazione a scapito di sacrificare individui che sono necessari per mantenere viva popolazioni. Tuttavia, quando esemplari morti sono disponibili per cause naturali questa tecnica fornisce un modo utile per raccogliere più informazioni di quanto sarebbe altrimenti disponibile. È importante notare il significato di questa tecnica per rimuovere con successo otoliti di storione utilizzano questo metodo alternativo. tecniche esistenti rimozione otoliti dalla regione dorsale della testa sono ancora utile e vantaggioso, ma non applicabile per storione. Utilizzando questa nuova tecnica per esporre la cavità del cervello dà anche l&#39;accesso e l&#39;applicabilità ad altre aree funzione neurale e la morfologia del cervello. Questa nuova tecnica può essere utile perutilizzare su altre specie di pesce con pareti cranio molto spesse o quando vi è una quantità limitata di pesce da utilizzare per l&#39;estrazione dell&#39;otolite.

Sturgeon (Acipenseridae) le popolazioni di tutto il mondo sono in calo da oltre un secolo a causa di impatti, tra cui la perdita di habitat, la frammentazione della popolazione, e la pesca eccessiva in modo tale che molte popolazioni sono protetti da leggi statali e federali 1,2. Agenzie di gestione in tutti gli USA e il mondo hanno identificato lo storione come specie bersaglio per il restauro e il recupero 1. Caratteristiche della popolazione (la crescita, il reclutamento, la dieta) della maggior parte delle popolazioni di storione sono stati studiati per comprendere meglio le caratteristiche di biologia e storia di vita di base 1. A causa della condizione protetta della maggior parte delle popolazioni di storione, la valutazione di età è difficile in piccole popolazioni minacciate Se concede nessun sacrificio o danni alle persone. L&#39;estrazione delle strutture ossee ai fini della stima dell&#39;età e analisi crescita è stata una tecnica comune 3 uso .Il delle sezioni raggi pinna per determinare l&#39;età di Sturgesu è diventato un metodo pratico non letale, ma la verifica della tecnica è stata limitata e l&#39;età può essere molto difficile da decifrare nei soggetti anziani (&gt; 15 anni) 4. I ricercatori hanno valutato l&#39;uso di più strutture ossee in storione per il loro uso nel convalidare l&#39;età di Golfo storione (Acipenser oxyrinchus desoti) 5, Atlantic Sturgeon (A. o. Oxyrinchus) 6, Storione Bianco (A. transmontanus) 7, e il Lago di Sturgeon (A. fluvescens) 8 ha concluso che il primo raggio della pinna pettorale marginale fornito l&#39;età più accurata, ma ha concluso la procedura invasiva per essere deleterio per gli individui più grandi. Attualmente, vi è lo sviluppo di utilizzare metriche forma otoliti di distinguere tra le specie di storione del Nord America, con un altissimo tasso di successo 9. Dettagli sulla processo di rimozione del otoliti con una incisione trasversale dorsale alla base del cranio è un comune otolite supplementareMetodo ction trovato tutta la letteratura e chiaramente dettagliato per Secor et al. (1991) 10. Tuttavia, questo metodo è difficile da implementare sulla storione che hanno crani molto spesse con otoliti che sono piccole e delicate e tenderebbero a rompersi. Un ulteriore revisione della letteratura mostra nessuna altra metodologia dettagliata del processo di rimozione otoliti per storione. L&#39;obiettivo di questo lavoro è stato quello di dettaglio un metodo alternativo di estrazione di otoliti da Sturgeon inserendo nella cavità del cervello dal lato 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

Estrazione dei otoliti (ossa dell&#39;orecchio) da pesci che hanno i crani molto spesse può essere difficile e richiede molto tempo. La pratica comune di fare una incisione trasversale verticale sulla parte superiore del cranio con una mano o sega elettrica possono danneggiare la dell&#39;otolite se non eseguite correttamente. Storioni (Acipenseridae) sono una famiglia, in particolare, che hanno un grande e grosso cranio. Un nuovo metodo di laboratorio che entra nella cavità cervello dal lato ventrale del pesce per esporre otoliti era più facile che altri metodi di estrazione otoliti in letteratura. Metodi recensiti in letteratura sono progettati per il campo e sono più efficienti a trattare grandi quantità di pesce rapidamente. Tuttavia, questa nuova tecnica è stato progettato per essere più adatto per un ambiente di laboratorio, quando il tempo non è pressante ed estrazione di successo di ciascun campione è critica. Il successo di trovare e rimuovere otoliti utilizzando questa tecnica è molto elevato e non compromette le struttura in qualsiasi modo. Questa tecnica alternativa è applicabile ad altre specie di pesci simili per l&#39;estrazione dei otoliti.

La dimensione adulta di storioni in tutto il mondo varia notevolmente ma la posizione degli otoliti è coerente. L&#39;uso di un coltello filetto tagliente è stato trovato per rimuovere il tessuto molle della bocca facilmente mostrato nella Figura 1D. Una sega elettrica osso è lo strumento preferito per l&#39;incisione laterale per esporre la cavità cerebrale in figura 1E. Tuttavia, l&#39;attenzione per bisettrice la linea mediana del cranio, serve una altrimenti i otoliti può essere schiacciato durante questo processo. Se questo processo viene eseguito correttamente, cavità cerebrale sarà esposto uniformemente senza danneggiare otoliti come mostrato nella Figura 2A. La materia cerebrale viene facilmente rimosso con l&#39;uso di pinze come in Figura 2B e trovare la cartilagine canale semicircolare che contiene il dell&#39;otolite è facilmente si trova alla base della cavità cerebrale come mostrato nella Figura 2C. La cartilagine trovato nel canale semicircolare è easglia rimosso dal otoliti con un paio di pinze con riferimento alla figura 2D. Questa procedura mostra correttamente la rimozione del otoliti storione e è un passo preliminare in un&#39;analisi più quantitativa della storia della vita del pesce. Esito successo può essere misurato dalla rimozione del otoliti senza rompere la struttura. tassi di determinazione dell&#39;età e crescita sono due metriche che possono essere valutati con l&#39;uso degli otoliti rimossi. Analisi Forma per la differenziazione delle specie è un altro parametro misurabile che può essere sviluppato dagli otoliti. <img alt="Figura 1" src="/files/ftp_upload/54316/54316fig1.jpg" /> Figura 1. Posizionamento di Sturgeon Carcassa per la rimozione Otolith. (A) Sturgeon Carcassa Vista Laterale per Misura e di pesatura, (B) verticale incisione per Separare Sturgeon testa da Bo<html> dy, (C) flip testa a Visualizza Bocca e ventrale lato della testa, (D) dei tessuti molli Rimosso per esporre ventrale laterale del cranio, (E) incisione laterale dalla punta del muso alla base del cranio <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54316/54316fig1large.jpg" target="_blank">Cliccate qui per vedere una versione più grande di questa figura.</a> <img alt="figura 2" src="/files/ftp_upload/54316/54316fig2.jpg" /> Figura 2. esame interno del cervello cavità per rimuovere otoliti. (A) del cervello Cavità esposti dopo testa è separata, (B) materia cerebrale Rimosso per esporre canali semicircolari, (C) Canale semicircolare Cartilagine rimossi, (D) Otolith Visto e rimosso da Canale semicircolare Cartilagine, (E) Shortnose Sturgeon Otolith54316 / 54316fig2large.jpg &quot;target =&quot; _ blank &quot;&gt; Clicca qui per vedere una versione più grande di questa figura.

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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.

Metodo alternativo di rimozione otoliti da 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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8.4K Views.  Great Lakes Science Center. The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses. 

Video: 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.

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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.

Sistemi di trattamento vegetale per la rimozione dei contaminanti associati alla tossicità dell&#39;acqua superficiale nell&#39;agricoltura e nella rottura urbana

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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.

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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.

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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.

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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.

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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.

Esempio di metodo di preparazione del microscopio elettronico di scansione e trasmissione per le appendici di Woodboring Beetle

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.

Estrazione di Cortisol da Sturgeon Fin e Matrici Jawbone

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.

Un metodo per quantificare gli artropodi 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.

Metodo elettrostatico per rimuovere il particolato organico dal suolo

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