Questa ricerca è stata sostenuta da Grant 105AS-13.2.3-BQ-B1 dal Bureau of animali e piante di ispezione sanitaria e la quarantena, il Consiglio per l&#39;Agricoltura, Executive Yuan e Grant 103-2313-B-197-002-MY3 dal Ministero della Scienza e della Tecnologia (MOST).

1. preparativi <ol><li> Fare 1 L di 50% sciroppo di zucchero. Sciogliere 1 kg saccarosio in 1 L DDH 2 O. </li><li> Preparare la soluzione piriprossifen (PPN) in BLD. Rendere 1,1 L di 10.000 ppm PPN soluzione madre e diluire 100 ml di soluzione PPN in 1 L sterilizzato DDH 2 O. Conservare a 4 ° C. </li><li> Diluire la soluzione madre PPN a concentrazioni finali di 0,1, 1, 10 e 100 mg / kg (ppm) nella BLD per il seguente esperimento. </li><li> Fare PPN-sciroppo (per il livello di colonia). Diluire il brodo PPN a concentrazioni finali di 10 e 100 ppm nel 50% sciroppo di zucchero per il seguente esperimento. </li><li> Il miele delle api allevamento. Nota: Qui, la posizione sperimentale è l&#39;apiario nazionale Ilan University (NIU), Yi-Lan City, Taiwan; (Coordinate GPS: N24.747278, E121.746200). <ol><li> Controllare miele ape (Apis mellifera L.) colonie settimanale per quantità di alimenti e mangimi con 1 litro di sciroppo di zucchero 50%, se necessario (l&#39;area di memorizzazione miele è vuoto). Definire un sanocolonia 9 fotogrammi di pettini api in ogni colonia con una regina che le uova normalmente. </li></ol></li><li> Preparare 100 ml di dieta larvale di base (BLD). Sciogliere 6% D-glucosio, fruttosio 6% e 1% di estratto di lievito in acqua deionizzata distillata sterilizzata (DDH 2 O) e integra con il 50% gelatina reale. Conservare a 4 ° C, ma non per più di 3 giorni. Nota: Pre-caldo a 35 ° C prima dell&#39;esperimento e consumare entro 3 giorni. </li></ol> 2. In Vivo metodo di alimentazione Nota: In vivo metodo di alimentazione è stato modificato da Hanley et al. 20 <ol><li> Il miele delle api selezione larve e l&#39;etichettatura. <ol><li> Inserire un escludi regina dividere 9 fotogrammi di una colonia sana in 4 frame e 5 fotogrammi limitando la regina alla sezione 4 telaio. Lasciare almeno un fotogramma vuoto nella sezione 4 telaio per la posa delle uova. </li><li> Dopo la regina deporre le uova per 1 giorno, Controllare i telai per la comparsa di uova e mantenere le uova all&#39;interno delle arnie per 72 ore (il 4 ° giorno) fino al 1 ° giorno di vita schiusa delle larve. Prendete una delle cornici contenenti 1-giorni di età le larve dei lavoratori (tratteggiata entro 24 ore) fuori dall&#39;alveare di prova a mano e togliere i lavoratori api dal telaio con una spazzola ape. </li><li> Coprire il telaio con un vetrino trasparente (Formato = lunghezza x larghezza x spessore = 29,7 millimetri x 21 mm x 0,1 mm) e inchiodare il vetrino trasparente sul bordo dei telai con puntine (Figura 1A). </li><li> Per ogni trattamento, selezionate casualmente 50 una larva di un giorno (il 4 ° giorno) e contrassegnare ciascuna cella covata utilizzando pennarelli permanenti sul vetrino trasparente (Figure 1A e 1B). Nota: scrivere l&#39;informazione di ogni trattamento sul telaio e scorrevole trasparente nonché utilizzando pennarelli permanenti per evitare la confusione tra diversi trattamenti. Rimuovere i vetrini segnalati e conservare per l&#39;in vivo &lt;/ Em&gt; alimentazione e osservazione. </li></ol></li><li> Nell&#39;alimentazione vivo. <ol><li> Aggiungere diverse concentrazioni di PPN-BLD (0,1, 1, 10 e 100 ppm) a ciascuna cella covata marcato pipettando al giorno 1, 2 e 3 con 10 microlitri, 10 microlitri e 20 uL, rispettivamente, secondo la quantità di aspirazione del età larve. Aggiungere la stessa quantità di BLD (senza PPN) ad un gruppo di controllo. In tal modo, la dose totale di PPN-BLD in ogni cella covata etichettati accumula 4, 40, 400, e 4000 pg. NOTA: Riconoscere le celle di covata etichettati dai vetrini trasparenti segnalati e rimuovere i vetrini trasparenti selezionati dopo l&#39;alimentazione. Utilizzare fresco BLD per l&#39;alimentazione per prevenire la pulizia delle celle di covata dalle api operaie. </li><li> Riportare i telai PPN-trattati nelle colonie originali per ulteriori osservazioni. NOTA: Ogni trattamento ha quattro ripete biologici in quattro colonie. </li></ol></li><li> L&#39;osservazione delle larve trattate al giorno 7. <ol><li> Per osservare °larve e PPN-trattati, restituire le diapositive trasparenti etichettati ai telai per registrare la mortalità e tassi di tappatura nelle celle di covata etichettati. </li></ol></li><li> L&#39;osservazione dei tassi di pupe e Eclosion trattati a giorno 13. <ol><li> Rimuovere la cera d&#39;api di celle di covata innevate. </li><li> Mettere le pinzette morbida capovolti nella cella di covata e serrare le pupe leggermente poi prendere pupe con delicatezza. </li><li> Trasferire il pupe in piastre da 24 pozzetti di coltura tissutale con un doppio strato di tessuto di laboratorio sotto ogni pozzetto. Registrare il danno e la mortalità durante il trasferimento di pupa. </li><li> Mantenere le piastre da 24 pozzetti di coltura tissutale in un incubatore a 34 ° C e 70% di umidità relativa fino emergere (circa 8 giorni). </li><li> Osservare e registrare le pupe e api mellifere emerse. </li></ol></li><li> statistica <ol><li> Calcolare i dati registrati e presente come media ± SD </li><li> Analizzare i dati utilizzando l&#39;analisi della varianza (ANOVA) da SAS e utilizzare il test minima differenza significativa (LSD) per analizzare le differenze tra due mezzi diversi trattamenti. Definire statisticamente significativo come P -value &lt;0.05. Lettere diverse nella stessa colonna della tabella mostrato un effetto significativo dall&#39;analisi statistica. </li></ol></li></ol> 3. Tossicità dei PPN a Colony Livello nella Beehive <ol><li> Impostare i gruppi di api. <ol><li> Inserire un escludi regina verticalmente dividere 9 fotogrammi di una colonia sana in 4 fotogrammi (parte A) e 5 fotogrammi (parte B), limitando la regina alla sezione 4 telaio. Lasciare almeno un fotogramma vuoto nella sezione 4-struttura per la deposizione delle uova. Nota: In altre escludi-regina del sulla parte superiore della parte della regina per evitare che la regina di muoversi tra le parti. </li><li> Dopo la regina deporre le uova per 1 giorno, controllare i telai per la comparsa di uova. Prendere i fotogrammi appropriati contenenti uova fuori dalla parte A a mano e togliere i lavoratori apidal telaio con una spazzola ape. </li><li> Coprire il telaio con scivolo trasparente e inchiodare il vetrino trasparente al bordo del telaio con puntine. </li><li> Selezionare casualmente 100 celle di covata contenenti uova e contrassegnare ciascuna cella covata utilizzando pennarello indelebile sul vetrino trasparente. Assegnare queste celle di covata 100 come gruppo 1. scrivere le informazioni di ciascun trattamento sul telaio e scorrevole trasparente con pennarello indelebile per evitare la confusione tra diversi trattamenti. </li><li> Riportare la cornice etichettati per parte A per 3 giorni e poi trasferire la regina parte B per deporre le uova. </li><li> Dopo 1 giorno, controllare i fotogrammi di parte B, scegliere una cornice adeguata ed etichetta 100 celle di covata contenenti uova come descritto al punto 3.1.4. Assegnare queste celle di covata 100 come gruppo 2. </li><li> Riportare la cornice etichettati per parte B per 3 giorni e poi trasferire la regina alla parte A per deporre le uova. </li><li> Ripetere lo scambio regina tra le parti A e B 6 volte di più e assegnare gruppi numericamente,rispettivamente (Figura 2). Ci dovrebbe essere totale di 9 gruppi. </li></ol></li><li> T miele reat colonia di api con PPN sciroppo di zucchero al giorno 13 (Figura 2). <ol><li> Aggiungere 1 L 50% PPN sciroppo di zucchero contenenti 10 o 100 ppm in una cassa di alimentazione ape plastica (L x L x H = 20 cm x 30 cm x 3,5 cm) e mettere la scatola in cima delle cornici nelle colonie sperimentali. Nota: Gruppo 1 non riceve il PPN a 13 giorni, come le celle di covata sono stati sigillati. </li><li> Alimentare il gruppo di controllo con 1 L 50% di sciroppo di zucchero (senza PPN) come descritto al punto 2.2. </li></ol></li><li> Conta 1 giorno di età larve e record come il tasso di schiusa dell&#39;uovo di 100 cellule marcate covata per ciascun gruppo a giorno 5 (Figura 2). Miele api uova di solito prendere 3 giorni per schiudersi in 0 giorni larve; quindi, controllare e etichettare 100 uova-contenenti celle di covata al giorno 1 e contare il numero di 1 giorno di età larve per ogni gruppo di giorno 5 per ottenere la percentuale di schiusa tasso. </li><li> CMONTAGGIO DEL celle di covata ricoperti e registrare come il tasso massimo larvale di 100 cellule marcate covata per ciascun gruppo a giorno 11 (Figura 2). 6 a 7 celle di covata larvale di un giorno saranno ricoperti con cera d&#39;api da parte dei lavoratori ape per le larve pupating. </li><li> Osservare pupe maturazione e registrare il tasso sfarfallamento di 100 cellule marcate covata per ciascun gruppo a giorno 17 (Figura 2). <ol><li> Rimuovere la cera d&#39;api di celle di covata capped e prendere pupe con morbidi pinzetta a punta delicatamente. Trasferire il pupe in un piastre di coltura tissutale a 24 pozzetti con un doppio strato di tessuto di laboratorio sotto ogni pozzetto. </li><li> Mantenere le piastre da 24 pozzetti di coltura tissutale in un incubatore a 34 ° C e 70% di umidità relativa fino emersione. </li><li> Osservare e registrare le pupe ed emerse le api per ogni gruppo fino Gruppo 9 (49 giorni sperimentali). Nota: Ogni trattamento ha quattro ripete biologici. </li></ol></li><li> statistica <ol><li> Calcolare i dati registrati e presenti come mediaSD ±. </li><li> Analizzare le differenze significative tra coppie di trattamenti (per esempio 0 ppm / 10 ppm, 10 ppm / 100 ppm e 0 ppm / 100 ppm) in ogni gruppo utilizzando t-test a due code di Student. Definire come statisticamente significativo se P -value &lt;0.05. </li></ol></li></ol>

<ol>
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Gli autori dichiarano di non avere interessi finanziari concorrenti.

Il metodo di deposizione delle uova queen-limitata e il metodo queen-scambio sono passaggi critici per la creazione di gruppi di api per le prove sul campo all&#39;interno di questo protocollo. Il metodo regina limitata ovodeposizione permette la sincronizzazione del ciclo di vita delle api. Di conseguenza, i ricercatori possono selezionare 1 giorno-vecchio larve della stessa età per il trattamento con diverse dosi di pesticidi. Per il metodo francese, lo scambio, la regina è stato scambiato tra la parte A (4 fotogram...

La presenza di pesticidi nell&#39;ambiente è uno dei problemi più gravi che l&#39;impatto della vita di un&#39;ape 1, 2, 3. Diversi studi hanno dimostrato la presenza di residui di pesticidi comuni in colonie di api mellifere e prodotti delle api. A Taiwan, l&#39;applicazione di pesticidi media era 11-12 kg / ha ogni anno (2005-2013). La quantità di pesticidi utilizzati a Taiwan è superiore a quella dei paesi dell&#39;UE, ei paesi dell&#39;America Latina 4, 5. In altre parole, l&#39;ambiente all&#39;apicoltura sta soffrendo gravi sollecitazioni di pesticidi, soprattutto a Taiwan e forse anche in altri paesi. Il miele ape mellifera è uno dei principali impollinatori nei sistemi agricoli 6 e produce anche prodotti pregiati come miele. Tuttavia, le api sono manifestazioniEd a vari pesticidi e questi pesticidi può essere riportato al alveari dopo foraggiamento sui fiori che sono stati spruzzati con pesticidi quando si raccolgono nettare e polline 7, 8. Essi inoltre possono essere esposti ai pesticidi dagli apicoltori stessi che mirano a controllare i problemi dei parassiti all&#39;interno delle arnie 9, 10, 11. Perché il miele delle api larve sono alimentati da api nutrici per il loro sviluppo, le larve, droni e perfino la regina può essere esposto a questi nettari pesticidi contaminati e polline 12. La tossicità dei vari pesticidi per le api deve essere affrontato 13. Molti sforzi sono stati fatti per valutare le emissioni di residui di pesticidi ambientali. Yang et al. testato l&#39;influenza del imidacloprid insetticidi neurotossici sullo sviluppo di miele delle api larve nelalveare e ha riferito che una dose sub-letali di imidacloprid ha provocato un comportamento associativo olfattiva delle api adulte 14. Inoltre, Urlacher et al. ha esaminato gli effetti sub-letali di una pesticidi organofosfati, clorpirifos, sulle prestazioni di apprendimento un miele di ape operaia in condizioni di laboratorio 15. Nel nostro studio precedente, abbiamo valutato l&#39;impatto di un regolatore di crescita degli insetti (IGR), piriprossifen (PPN), su api da miele larvali 16. In questo documento, presentiamo metodi per valutare gli effetti chimici sullo sviluppo delle api. Miele metodi di alimentazione ape sono stati descritti ed applicati sia singole api mellifere o ad una colonia. In un primo momento, abbiamo provato diverse concentrazioni di pesticidi contaminato dieta larvale di base (BLD) di larve nelle colonie per valutare l&#39;impatto del pesticida sulle singole api mellifere in vivo. Si è quindi proceduto a simulare la Condit naturaliioni del pesticida utilizzando lo sciroppo di pesticidi contaminati all&#39;interno di alveari. In questo metodo, PPN, che è ampiamente usato contro insetti parassiti 17 ed è dannoso per lo sviluppo di miele api larve e pupe 16, 18, 19, sarà un indicatore per rappresentare l&#39;effetto negativo del pesticida nel campo.

<table border="0" cellpadding="0" cellspacing="0" width="887">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Honey bee box</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">W1266</td>
			<td style="width:297px;">Honeybees rearing</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Queen excluder (between frames)</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1575</td>
			<td style="width:297px;">Queen limitation&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Queen excluder (on top )</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1566</td>
			<td style="width:297px;">Queen limitation on top&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Bee brush</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>W1414</td>
			<td style="width:297px;">clean the bees on frame gently</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:193px;">Bee feeder</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>P0219</td>
			<td style="width:297px;">feed sugar syrup to colony</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;">Transparent slide</td>
			<td style="width:243px;">Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40)</td>
			<td>1139</td>
			<td style="width:297px;">Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size= Length*Width*thick= 29.7mm*21mm*0.1mm)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">24 well tissu culture plate</td>
			<td style="width:243px;">Guangzhou Jet Bio-Filtration Co., Ltd</td>
			<td style="width:155px;">TCP011024</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Autoclave</td>
			<td style="width:243px;">Tomin medical equipmenco., LTD.</td>
			<td style="width:155px;">TM-321</td>
			<td style="width:297px;">Make sterilized distilled deionized water (ddH2O)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">P20 pipetman</td>
			<td style="width:243px;">Gilson</td>
			<td style="width:155px;">F123600</td>
			<td style="width:297px;">Add PPN into bee larval food pool</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:193px;">Incubator&#160;</td>
			<td style="width:243px;">Yihder Co., Ltd.</td>
			<td style="width:155px;">LE-550RD</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Kimwipes</td>
			<td style="width:243px;">COW LUNG INSTRUMENT CO., LTD</td>
			<td style="width:155px;">KCS34155</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Royal jelly</td>
			<td style="width:243px;">National Ilan University (NIU)</td>
			<td style="width:155px;">NIU</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(+)-Glucose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">G8270</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(-)-Fructose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">F0127</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Yeast extract</td>
			<td style="width:243px;">CONDA, pronadisa</td>
			<td style="width:155px;">1702</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">Sucrose</td>
			<td style="width:243px;">Taiwan sugar coporation</td>
			<td style="width:155px;">E01071010</td>
			<td style="width:297px;">Make sugar syrup for bee food</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Pyriproxyfen (11%)</td>
			<td style="width:243px;">LIH-NUNG CHEMICAL CO.. LTD.</td>
			<td style="width:155px;">Registration No. 1937</td>
			<td style="width:297px;">Insect growth regulator (IGR) used in the experiment</td>
		</tr>
	</tbody>
</table>

evaluating the effect of environmental chemicals on honey bee development from the individual to colony level

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be brought back to the beehive after the bees have foraged on flowers that have been sprayed with pesticides. Pesticide contaminated food can be exchanged between workers which then feed larvae and therefore can potentially affect the development of honey bees. Thus, residual pesticides in the environment can become a chronic damaging factor to honey bee populations and gradually lead to colony collapse. In the presented protocol, honey bee feeding methods are described and applied to either an individual honey bee or to a colony. Here, the insect growth regulator (IGR) pyriproxyfen (PPN), which is widely used to control pest insects and is harmful to the development of honey bee larvae and pupae, is used as the pesticide. The presenting procedure can be applied to other potentially harmful chemicals or honeybee pathogens for further studies.

Per la prova sul campo miele ape, una regina era limitato alla sezione 4-struttura per la deposizione delle uova. Questa fase potrebbe aumentare la densità covata in un frame e facilitare osservazioni successive. Ogni trattamento è stato etichettato, e lo sviluppo delle api stato chiaramente osservato attraverso un vetrino trasparente. In vivo alimentazione di PPN-BLD al miele api larve dell&#39;alveare è stata eseguita per valutare con precisione l&#39;influenza della PPN su...

Watch this Scientific Journal Video about Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level at JoVE.com

Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

Qui vi presentiamo un metodo per alimentare antiparassitario alimenti contaminati sia un individuo ape e una colonia alveare. La procedura valuta l&#39;effetto dei pesticidi sulle singole api mellifere in vivo l&#39;alimentazione della dieta larvale di base e anche la condizione naturale della colonia dell&#39;alveare.

Valutare l&#39;effetto delle sostanze chimiche ambientali su Honey Bee sviluppo dall&#39;individuo alla colonia Livello

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be ...

evaluating-effect-environmental-chemicals-on-honey-bee-development

Universidad San Sebastian

Research

JoVE Journal

Environment

9.0K Views.  National Ilan University.  Qui vi presentiamo un metodo per alimentare antiparassitario alimenti contaminati sia un individuo ape e una colonia alveare. La procedura valuta l'effetto dei pesticidi sulle singole api mellifere in vivo l'alimentazione della dieta larvale di base e anche la condizione naturale della colonia dell'alveare. 

Video: Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. 
In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

Automated Gel Selection Size per migliorare la qualità delle Biblioteche di sequenziamento di nuova generazione Preparato da campioni di acqua ambientale

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

Ricerca

Ambiente

Exploring the Effects of Atmospheric Forcings on Evaporation: Experimental Integration of the Atmospheric Boundary Layer and Shallow Subsurface

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study evaporation under various surface boundary conditions to improve our current understanding and characterization of this multiphase phenomenon as well as to validate numerical heat and mass transfer theories that couple Navier-Stokes flow in the atmosphere and Darcian flow in the porous media. Experimental data were collected using a unique soil tank apparatus interfaced with a small climate controlled wind tunnel. The experimental apparatus was instrumented with a suite of state of the art sensor technologies for the continuous and autonomous collection of soil moisture, soil thermal properties, soil and air temperature, relative humidity, and wind speed. This experimental apparatus can be used to generate data under well controlled boundary conditions, allowing for better control and gathering of accurate data at scales of interest not feasible in the field. Induced airflow at several distinct wind speeds over the soil surface resulted in unique behavior of heat and mass transfer during the different evaporative stages.

A protocol for the design and construction of a soil tank interfaced to a small climate controlled wind tunnel to study the effects of atmospheric forcings on evaporation is presented. Both the soil tank and wind tunnel are instrumented with sensor technologies for the continuous in situ measurement of environmental conditions.

Esplorare gli effetti dell&#39;inquinamento atmosferico forcing per evaporazione: Integrazione sperimentale dello strato atmosferica Boundary e Subsurface Shallow

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study ...

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted through ion exchange. The resultant production bleed water (PBW) contains contaminants such as arsenic and other heavy metals. Samples of PBW from an active ISR uranium facility were treated with cupric oxide nanoparticles (CuO-NPs). CuO-NP treatment of PBW reduced priority contaminants, including arsenic, selenium, uranium, and vanadium. Untreated and CuO-NP treated PBW was used as the liquid component of the cell growth media and changes in viability were determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay in human embryonic kidney (HEK 293) and human hepatocellular carcinoma (Hep G2) cells. CuO-NP treatment was associated with improved HEK and HEP cell viability. Limitations of this method include dilution of the PBW by growth media components and during osmolality adjustment as well as necessary pH adjustment. This method is limited in its wider context due to dilution effects and changes in the pH of the PBW which is traditionally slightly acidic&#160;however; this method could have a broader use assessing CuO-NP treatment in more neutral waters.

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

Rimozione di Oligoelementi per Cupric nanoparticelle di ossido di uranio da<em&gt; In Situ</em&gt; Recupero Bleed Acqua e il suo effetto sulla vitalità cellulare

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

Un sistema di esposizione di flusso-through per valutare sospeso Sedimenti Effetti sulla vita acquatica

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

Estrazione di pesticidi di organochlorato da pellicole di plastica e analisi di tipo plastico

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

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.

Preparazione di MAURIZIOBANDIERIV e DMDTAV mediante DMAV per applicazioni ambientali: sintesi, purificazione e conferma

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

The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food safety and to extend the shelf life of meat and meat products, the use of either synthetic or natural preservatives have been applied to control and eliminate foodborne bacteria. A growing interest in the application of natural food additives for meat has increased. Microorganisms, such as Escherichia coli, contaminate meat and meat products, causing foodborne illnesses. Therefore, it is necessary to improve the meat conservation process. However, the use of essential oils when the meat is being dried has not been deeply studied. In this regard, there is an opportunity to increase the value of dried meat and reduce the risk of foodborne illnesses by applying essential oils during the drying process. In this protocol, we present a novel method of applying thyme essential oil (TEO) during meat drying, specifically in vapor form directly in a drying chamber. For evaluation, we use Minimal Inhibitory Concentration (MIC) to detect the number of harmful bacteria in the treated samples compared to raw samples. The preliminary results show that this method is a viable and alternative option to synthetic preservatives and that it significantly reduces microbial load in dried meat.

Microorganisms such as Escherichia coli that contaminate meat products cause foodborne illnesses. The use of essential oils in the meat drying process has not been deeply studied. Here, we present a novel method of applying thyme essential oil to meat during drying to reduce the microbial load in dried meat.

L'effetto dell'applicazione dell'olio essenziale di timo sulla carica microbica durante l'essiccazione della carne

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food ...

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species.
This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

In this work, we describe an acute, chronic and multigenerational bioassay to study the effects of single and combined stressors on the Turquoise killifish Nothobranchius furzeri. This protocol is designed to study life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Protocollo per acuto e prove di ecotossicità cronica del turchese Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity ...

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee (Apis mellifera) Workers

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired glands that are located in the head of the bee are highly sensitive to the quantity and quality of pollen and pollen substitutes that the nurse bee consumes. The glands get smaller when nurses are fed deficient diets and are large when they are fed complete diets. Because nurse hypopharyngeal gland size is a robust indicator of nurse nutrition, it is essential that those studying honey bee nutrition know how to measure these glands. Here, we provide detailed methods for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal glands. We present comparisons of unstained and stained tissue and data that were used to study the impact of pollen on gland size. This method has been used to test how diet impacts hypopharyngeal gland size but has further use for understanding the role of these glands in hive health.

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee Apis mellifera Workers

Hypopharyngeal gland acinus size is a robust measure of nurse honey bee nutrition. Here, we provide a detailed protocol for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal gland acini.

Formato di Acinus ghiandola Hypopharyngeal misurazione in lavoratori ape del miele (mellifera di Apis)

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

Composizione e analisi della distribuzione di Bioaerosols in diverse condizioni ambientali

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...