Esta pesquisa foi apoiada por Grant 105AS-13.2.3-BQ-B1 do Bureau of Animal e Inspeção de Saúde de Plantas e Quarentena, o Conselho de Agricultura, Yuan Executivo e Grant 103-2313-B-197-002-MY3 do Ministério da Ciência e Tecnologia (MOST).

1. Preparações <ol><li> Adicione 1 L de 50% de xarope de açúcar. Dissolve-se 1 kg de sacarose em 1 L de ddH 2 O. </li><li> Preparar a soluo de piriproxifeno (PPN) em BLD. Adicione 1,1 L de solução de estoque de 10000 ppm PPN e diluir 100 mL de solução de PPN em 1 L esterilizados ddH 2 O. Armazenar a 4 ° C. </li><li> Dilui-se a solução estoque PPN para concentrações finais de 0,1, 1, 10 e 100 mg / kg (ppm) no BLD para a experiência seguinte. </li><li> Faça PPN-xarope (para o nível de colônia). Dilui-se a estoque PPN para concentrações finais de 10 e 100 ppm em 50% de xarope de açúcar para a experiência seguinte. </li><li> Mel criação de abelha. Nota: Aqui, a localização experimental é o apiário Universidade Nacional Ilan (NIU), Yi-Lan City, Taiwan; (Coordenadas GPS: N24.747278, E121.746200). <ol><li> Verificar mel de abelha (Apis mellifera L.) colónias semanal para a quantidade de alimentos e alimentos para animais com 1 litro de 50% de xarope de açúcar, se necessário (a área de armazenamento de mel está vazio). Definir uma saudávelcolónia como 9 quadros de favos de mel de abelha, em cada colónia com uma rainha postura dos ovos normalmente. </li></ol></li><li> Preparar 100 mL de dieta da larva de base (BLD). Dissolve-se 6% de D-glucose, 6% de frutose, e 1% de extracto de levedura em água desionizada destilada esterilizada (ddH2O) e completar com 50% a geleia real. Armazenar a 4 ° C, mas não mais de 3 dias. Nota: Pré-quente a 35 ° C antes da experiência e utilizar no prazo de 3 dias. </li></ol> 2. In Vivo método de alimentação Nota: In vivo método de alimentação foi modificado a partir de Hanley et al. 20 <ol><li> Mel de abelha selecção larvas e rotulagem. <ol><li> Inserir um excluidora para dividir 9 quadros de uma colónia saudável para 4 quadros e quadros 5, limitando a rainha para a secção de armação 4. Deixe pelo menos uma moldura vazia na seção 4 moldura para colocar ovos. </li><li> Após a rainha de pôr ovos por 1 dia, Veja os quadros para o aparecimento de ovos e manter os ovos no interior das colmeias de 72 h (a 4 ° dia) até 1 dia de idade larvas eclodem. Tomar um dos quadros contendo larvas trabalhador com 1 dia de idade (tracejado dentro de 24 h) para fora a partir do ramo de teste pela mão e remover os trabalhadores da abelha do mel a partir da estrutura com uma escova de abelha. </li><li> Cobrir o quadro com uma lâmina transparente (tamanho = comprimento x largura x espessura = 29,7 milímetros x 21 mm x 0.1 mm) e pregar a lâmina transparente na extremidade da armação com percevejos (Figura 1A). </li><li> Para cada tratamento, seleccionar aleatoriamente 50 um dia de idade, as larvas (a 4 ° dia) e marcar cada célula ninhada utilizando canetas permanentes na transparente corrediça (Figuras 1A e 1B). Nota: Adicione a informação de cada tratamento sobre a armação e corrediça transparente assim usando canetas permanentes para evitar a confusão entre os diferentes tratamentos. Remova os slides marcados e manter para o in vivo &lt;/ Em&gt; alimentação e observação. </li></ol></li><li> Na alimentação in vivo. <ol><li> Adicionar diferentes concentrações de PPN-BLD (0,1, 1, 10 e 100 ppm) para cada célula ninhada marcado por pipetagem no dia 1, 2 e 3 com 10 mL, 10 mL e 20 mL, respectivamente, de acordo com a quantidade de admissão do idade larvas. Adicionar a mesma quantidade de BLD (sem PPN) a um grupo de controlo. Desse modo, a dose total de PPN-BLD em cada célula de cria marcado acumula-se 4, 40, 400, e 4000 pg. NOTA: Reconhecer as células de cria rotulados pelas lâminas transparentes marcados e remover as lâminas transparentes marcados após a alimentação. Use fresco BLD para alimentar para prevenir a limpeza das células de cria pelas abelhas obreiras. </li><li> Devolver os quadros-tratados PPN para as colónias originais para observações adicionais. NOTA: Cada tratamento tem quatro repetições biológicas em quatro colônias. </li></ol></li><li> Observação de larvas tratadas no dia 7. <ol><li> Para observar the PPN-tratada larvas, retornar os slides transparentes etiquetados para os quadros para gravar a mortalidade e taxa máxima nas células de cria rotulados. </li></ol></li><li> Observação de taxas de eclosão e de pupa tratados no dia 13. <ol><li> Remova a cera de abelha de células criação selada. </li><li> Coloque as pinças macio derrubado na célula de cria e prender o pupas muito ligeiramente em seguida, tomar pupas suavemente. </li><li> Transferir as pupas em placas de 24 cavidades de cultura de tecido com uma camada dupla de tecidos de laboratório sob cada poço. Grave o dano e mortalidade durante a transferência de pupa. </li><li> Manter as placas de 24 cavidades de cultura de tecidos em uma incubadora a 34 ° C e 70% de humidade relativa até emergência (cerca de 8 dias). </li><li> Observar e registar a pupas e abelhas surgiram. </li></ol></li><li> Estatisticas <ol><li> Calculam-se os dados gravados e presente como uma média ± SD </li><li> Analisar os dados utilizando análise de variância (ANOVA), SAS e usar o teste de diferença mínima significativa (LSD) para analisar as diferenças entre os dois meios de diferentes tratamentos. Definir estatisticamente significativa como P -valor &lt;0,05. Letras diferentes na mesma coluna da tabela mostraram um efeito significativo, a análise estatística. </li></ol></li></ol> 3. A toxicidade de PPN na Colony Nível em colmeia <ol><li> Configure os grupos de abelhas. <ol><li> Inserir um excluidora verticalmente para dividir 9 quadros de uma colónia saudável para 4 quadros (Parte A) e 5 quadros (Parte B), enquanto que limitam a rainha para a secção de armação 4. Deixar, pelo menos, um quadro vazio na secção 4-quadro para a postura dos ovos. Nota: Dito de outra rainha Excluder o em cima da parte rainha para evitar a rainha de se mover entre as partes. </li><li> Após a rainha de pôr ovos por 1 dia, verifique os quadros para o aparecimento de ovos. Pegue os quadros apropriados contendo ovos para fora da parte A com a mão e retirar os trabalhadores da abelha do mela partir da moldura com uma escova de abelha. </li><li> Cobrir o quadro com corrediça transparente e pregar a lâmina transparente para a borda do quadro com tachas. </li><li> Aleatoriamente selecionar 100 células de cria contendo ovos e marcar cada célula de cria usando marcador permanente sobre a lâmina transparente. Atribuir estes 100 células de cria como Grupo 1. Adicione a informação de cada tratamento sobre a armação e corrediça transparente usando o marcador permanente para evitar a confusão entre os diferentes tratamentos. </li><li> Devolver o quadro rotulado a parte A, durante 3 dias e depois transferir a rainha a parte B para colocar ovos. </li><li> Após 1 dia, veja os quadros de parte B, escolher uma estrutura apropriada e rotular 100 células de cria contendo ovos, tal como descrito no passo 3.1.4. Atribuir esses 100 células de cria como Grupo 2. </li><li> Devolver o quadro rotulado com a parte B por 3 dias e, em seguida, transferir a rainha à parte A para pôr ovos. </li><li> Repetir a troca rainha entre a parte A e parte B mais 6 vezes e atribuir grupos numericamente,respectivamente (Figura 2). Deve haver total de 9 grupos. </li></ol></li><li> T mel reat colónia de abelhas com PPN xarope de açúcar no dia 13 (Figura 2). <ol><li> Adicionar 1 L 50% PPN xarope de açúcar contendo 10 ou 100 ppm em uma caixa de plástico alimentador de abelha (W x L x H = 20 cm x 30 cm x 3,5 cm) e colocar a caixa no topo dos quadros nas colónias experimentais. Nota: O Grupo 1 não receber o PPN em 13 dias, como as células de cria foram selados. </li><li> Alimentar o grupo de controlo com 1 L de 50% de xarope de açúcar (não PPN) como descrito na etapa 2.2. </li></ol></li><li> Contagem de larvas de um dia de idade e ficha como a taxa de eclosão de ovos de 100 células de cria marcadas para cada grupo no dia 5 (figura 2). Mel ovos de abelhas costumam levar 3 dias para eclodir em 0 larvas dia; portanto, verificar e rotular 100 ovos células contendo ninhada no dia 1 e contar o número de larvas de um dia de idade para cada grupo no dia 5 para se obter a percentagem de taxa de eclosão. </li><li> Count as células criação selada e ficha como a taxa de nivelamento larvar de 100 células de cria marcadas para cada grupo no dia 11 (Figura 2). 6 a 7 de células de cria larval dia de idade serão cobertas com cera de abelha por trabalhadores de abelhas para pupating larvas. </li><li> Observar pupas maturação e registar a taxa de eclosão de 100 células de cria marcadas para cada grupo no dia 17 (Figura 2). <ol><li> Remova a cera de abelha de células criação selada e tomar pupas com uma pinça com ponta macia suavemente. Transferir as pupas para uma placas de cultura de tecidos de 24 cavidades com uma camada dupla de tecidos de laboratório sob cada poço. </li><li> Manter as placas de 24 cavidades de cultura de tecidos em uma incubadora a 34 ° C e 70% de humidade relativa até emergência. </li><li> Observar e registar a pupas e emergiu abelhas para cada grupo até Grupo 9 (49 dias experimentais). Nota: Cada tratamento tem quatro repetições biológicas. </li></ol></li><li> Estatisticas <ol><li> Calculam-se os dados gravados e presente como a mia± SD. </li><li> Analisar as diferenças significativas entre os pares de tratamentos (por exemplo, 0 ppm / 10 ppm, 10 ppm / 100 ppm e 0 ppm / 100 ppm) em cada grupo através da utilização de duas caudas teste t de Student. Definir como estatisticamente significativa se P -valor &lt;0,05. </li></ol></li></ol>

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Os autores declaram que não têm interesses financeiros concorrentes.

O método de postura de ovos limitados-rainha e método de permuta rainha são etapas críticas para a criação de grupos de abelhas para testes de campo dentro deste protocolo. O método de postura de ovos limitados-rainha permite a sincronização do ciclo de vida das abelhas de mel. Consequentemente, os pesquisadores podem selecionar um dia de idade larvas da mesma idade para o tratamento com diferentes doses de pesticidas. Para o método de permuta rainha, a rainha foi trocada entre a parte A (4 frames) e B (5 quad...

A presença de pesticidas no meio ambiente é um dos problemas mais graves que os impactos a vida de uma abelha do mel 1, 2, 3. Vários estudos têm demonstrado a presença de resíduos de pesticidas comum em colónias de abelhas e produtos das abelhas. Em Taiwan, a aplicação média de pesticidas foi 11-12 kg / ha a cada ano (de 2005 a 2013). A quantidade de pesticidas utilizados em Taiwan é maior do que a dos países da UE e os países da América Latina 4, 5. Em outras palavras, o ambiente apícola está sofrendo sério estresse de pesticidas, especialmente em Taiwan e possivelmente em outros países. O mel de abelha Apis mellifera é um dos principais polinizadores em sistemas agrícolas 6 e também produz produtos valiosos, como mel. No entanto, as abelhas são exposed para vários pesticidas e estes pesticidas podem ser trazidos de volta para colmeias depois das pastagens em flores que foram pulverizados com pesticidas ao recolher néctar e pólen 7, 8. Eles também podem ser expostos a pesticidas pelos apicultores-se com o objetivo de controlar os problemas de pragas dentro das colmeias 9, 10, 11. Porque as larvas de abelhas de mel são alimentados por abelhas enfermeira para o seu desenvolvimento, larvas, parasitas e mesmo a rainha podem ser expostos a estas néctares contaminados com pesticidas e pólen 12. A toxicidade de vários pesticidas para as abelhas precisa ser tratada 13. Muitos esforços têm sido feitos para avaliar as questões de resíduos de pesticidas ambientais. Yang et al. testado a influência do inseticida imidacloprid neurotóxicos no desenvolvimento de larvas de abelhas mel nocolmeia e relataram que uma dose sub-letal de imidacloprid resultou num comportamento associativo olfactiva das abelhas adultas 14. Além disso, Urlacher et al. examinaram os efeitos sub-letais de um dos pesticidas organofosforados, clorpirifos, sobre o desempenho de aprendizagem de um trabalhador da abelha do mel, em condições laboratoriais 15. No nosso estudo anterior, foi avaliado o impacto de um regulador de crescimento de insectos (IGR), piriproxifeno (PPN), em abelhas de mel larvais 16. Neste artigo, apresentamos métodos para avaliar os impactos químicos sobre o desenvolvimento das abelhas. métodos de alimentação de abelhas de mel foram descritos e aplicada quer abelhas produtoras de mel individuais ou a uma colónia. No primeiro, testaram-se diferentes concentrações de dieta da larva básica contaminada-pesticida (BLD) em larvas nas colónias para avaliar o impacto do pesticida em abelhas de mel individuais in vivo. Passamos então para simular o Condit naturaisiões do pesticida usando xarope contaminado-pesticida dentro de colmeias. Neste método, PPN, que é amplamente utilizado contra pragas de insectos 17 e é prejudicial para o desenvolvimento de larvas e pupas da abelha do mel 16, 18, 19, será um indicador para representar o efeito negativo do pesticida no 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.

Para o teste de campo da abelha do mel, uma rainha foi limitado à secção 4-quadro para a postura dos ovos. Este passo poderia aumentar a densidade de ninhada em um quadro e facilitar observações subsequentes. Cada tratamento foi rotulado, e desenvolvimento das abelhas foi claramente observado através de um slide transparente. Na alimentação in vivo de PPN-BLD para as larvas de abelhas na colmeia foi realizada para avaliar com precisão a influência de PPN sobre...

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

Apresentamos aqui um método para alimentar pesticida alimentos contaminados se tanto a um indivíduo abelha do mel e uma colónia colmeia. O procedimento avalia o efeito pesticida em abelhas de mel individuais por alimentação in vivo de dieta da larva de base e também na condição natural da colónia colmeia.

Avaliando o efeito de substâncias químicas ambientais em Honey Bee Desenvolvimento do indivíduo para Colony Nível

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

Research

JoVE Journal

Environment

9.0K visualizações.  National Ilan University.  Apresentamos aqui um método para alimentar pesticida alimentos contaminados se tanto a um indivíduo abelha do mel e uma colónia colmeia. O procedimento avalia o efeito pesticida em abelhas de mel individuais por alimentação in vivo de dieta da larva de base e também na condição natural da colónia colmeia. 

Vídeo: 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 seleção de tamanho para melhorar a qualidade das bibliotecas de sequenciação de próxima geração Preparado a partir de amostras de água do ambiente

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

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.

Explorando os efeitos de forçantes atmosféricas sobre evaporação: Integração Experimental da Camada Limite Atmosférica e Raso Subsurface

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.

A remoção do Trace Elements por Óxido de Cupric nanopartículas de urânio<em&gt; In Situ</em&gt; Recuperação sangro água e seu efeito na viabilidade celular

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.

Um Sistema de Exposição Fluxo através de sedimentação de Efeitos Avaliando suspensos sobre a vida aquática

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.

Extração de Pesticidas Organoclorados de Pelotização de Plástico e Análise de Tipo de Plástico

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.

Preparação de DMMTAV DMDTAV usando DMAV para aplicações ambientais: síntese, purificação e confirmação

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.

O efeito da aplicação do óleo essencial de tomilho na carga microbiana durante a secagem da 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.

Protocolo para aguda e crônica de ensaios toxicológicos do turquesa 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.

Medição de hipofaringe glândula Acinus tamanho em trabalhadores de abelhas (Apis mellifera)

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.

Composição e análise da distribuição de Bioaerossóis sob diferentes condições ambientais

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