Name: laboratory rearing of stable flies and other muscoid diptera
Uploaded: 2018-08-03T12:30:00.000+00:00
Duration: 7 min
Description: Watch this Scientific Journal Video about Laboratory Rearing of Stable Flies and Other Muscoid Diptera at JoVE.com

Gostaríamos de agradecer a Anthony Weinhold e os inúmeros estudantes que trabalharam connosco ao longo dos anos para suporte técnico, bem como sugestões para melhorar nosso procedimentos de criação de inseto.

1. ovo coleção (Figura 1)
<ol><li>Preparar a incitar a taça, coloque uma das extremidades do pano ~ 500 mL copo cheio de água morna (~ 40 ° C). Sobrepõem-se os lados do copo e fixe com um elástico. Dobre a ponta solta do pano volta por cima da taça.</li>
	<li>Lugar Copa a incitar na gaiola de moscas de 8 – 10 dias de idade para as mulheres Gravid de ~ 2 h. estável voa será ovipositar no pano.</li>
	<li>Remova a incitar a taça de ovos gaiola e enxaguar fora o pano egging em uma panela pequena com lavagem de garrafa.</li>
</ol>2. prepare Pan criação Larval
<ol><li>
Preparar o meio (quantidades para 1 pan de criação)
	<ol>
<li>Combinar o farelo de trigo (500 g), aparas de madeira (200 g) e farinha de peixe (115 g) em panela de prato plástico 10L e misture bem.</li>
		<li>Adicionar o bicarbonato de amónio (50 g) de água (~ 25 ° C, 1600 mL) e misture até dissolver.</li>
		<li>Adicione a solução de água de ingredientes secos e misture para não garantir nenhum material unwetted. Nível médio em panela mas não comprimir.</li>
	</ol>
</li>
	<li>
Adicione os ovos de mosca estáveis
	<ol>
<li>Faça um sulco raso em média o comprimento da bandeja.</li>
		<li>Depósito de 1 mL de ovos de mosca estáveis (~ 8.000 ovos) no sulco com pipeta de ovo (Figura 2). Nota: Pipeta de ovo feita por um corte de uma pipeta graduada plástica no ponto desejado, inserir 100 mesh tela e colagem pipeta junta novamente.</li>
		<li>Cubra os ovos com uma fina camada do meio para evitar a dessecação.</li>
		<li>Cubra a panela com fronha, fechar com uma faixa de borracha e etiqueta. Colocar panelas no quarto larval (23 ± 2 ° C, 30 – 50% RH e 12:12 [L:D] h fotoperíodo). Nota: Desenvolvimento Larval leva de 10 a 14 dias do tempo de oviposição.</li>
	</ol>
</li>
</ol>3. processamento pupa, as larvas se mover para a borda do meio 7 – 9 dias após a oviposição e Pupariate por dias 13 ou 14
<ol><li>Casulos de colher, localizados principalmente em torno das bordas da panela sob a crosta que se forma na superfície do meio, para fora e colocado em uma panela limpa.</li>
	<li>Encha a panela, com casulos, ½ a ¾ cheio de água. Separar grupos de casulos e médio. Componentes de farelo e proteína de trigo do meio de afundam-se enquanto pupários > 1 dia de idade flutuador19.</li>
	<li>Remova materiais flutuantes, casulos e alguns componentes médios com uma peneira e lavagem através de uma série de peneiras (5 #, 7 #, #12, #20) para remover o restante médio. A peneira #12 recolhe o casulo e a #20 evita que resíduos sólidos entrando pelo ralo. Casulos de lavagem entre peneiras com um pulverizador de pia.</li>
	<li>Casulos de enxágue, em peneira #12, em uma panela limpa. Encha a panela com ½ cheio de água. Despeje pupários flutuante em um coador e deixe escorrer o excesso de água.</li>
	<li>Transferência drenado pupários para tela de secagem com o ventilador e deixar até secar (Figura 3).</li>
</ol>4. pupa prateleira, um método alternativo para a coleta de pupários é com uma prateleira Pupal18
Nota: A prateleira é feita de um pedaço de plástico corte da extremidade de uma panela de prato de 10 L (10,2 cm de altura x 10,2 cm de largura x 31,9 cm de comprimento).
<ol><li>Prepare o suporte conforme descrito no ponto 2.1. Cone médio em panela de ~2.5 cm no fundo uma ponta de ~7.5 cm profundo no outro.</li>
	<li>
Coloque a prateleira no meio na parte rasa da criação pan e fita para evitar larvas se rastejando ao lado da prateleira (Figura 4).
	<ol>
<li>Saturar a esponja (~14.5 x 9.0 x 4.5 cm3, celulose regenerada) e cobrir com um pano embebido de água (30.5 x 42 cm2, algodão) e lugar na prateleira de cerca de meia polegada do meio.</li>
	</ol>
</li>
	<li>
Adicione os ovos, conforme descrito no ponto 2.2 e proceder como anteriormente.
	<ol>
<li>Verifique panelas diariamente para garantir que a esponja permaneça úmida.</li>
	</ol>
</li>
	<li>Casulos de enxaguamento da plataforma, esponja e pano em um prato limpo panela e despeje no coador, 14 dias após a oviposição.</li>
</ol>5. controle de qualidade
<ol><li>Pese todos os casulos produzidos no pan (peso total)..</li>
	<li>Isolar 100 casulos, pesar e colocado em uma placa de Petri de 9 cm.</li>
	<li>Contagem e sexo adultos ~ 5 a 8 dias mais tarde para determinar a taxa de surgimento e proporção sexual. Loja de placas de Petri com adultos surgiu em um freezer para a contagem em uma data posterior, se necessário. Moscas são sexadas pela forma de seus olhos e a largura da placa fronto-orbital20 ou genitália em pequeno aumento.</li>
	<li>Peso total de registro, peso de 100 casulos e número de surgiu adultos machos e fêmeas.</li>
</ol>6. preparação de sangue
<ol><li>Colete o sangue fresco de bovino de um matadouro local em baldes de 19 L contendo 70 g de citrato de sódio tribásico dihidrato em 500 mL de água. Mexa vigorosamente por 5-10 min de sangue e voltou para o laboratório.</li>
	<li>Sangue de tensão através de uma peneira para remover os coágulos e particionado em recipientes de 2 L, etiqueta com a data de colheita e armazenamento em um freezer (-20 ° C).</li>
	<li>Precisava de 2 dias antes do recipiente retire do congelador e coloque na geladeira. Sangue pode ser usado por ~ 2 semanas uma vez descongelados e armazenados no freezer por até 1 ano.</li>
</ol>7. adulta manutenção
<ol><li>Prepare as gaiolas, fundo de gaiola linha com papel pardo para facilitar a limpeza.</li>
	<li>Coloque 50 g de pupas de mosca secas estáveis (~ 3.500) em cada gaiola. Eclose adultos algumas horas a alguns dias após o processamento pupal.</li>
	<li>Lugar um sangue fresco embebido guardanapo feminino em cima de cada gaiola para alimentação. Começam a alimentar-se dentro de 24 h de adulto eclosão e repetir diariamente, até um dia antes da oviposição (9-10 dias após a emergência).</li>
	<li>Após a oviposição (secção 1), coloque a gaiola no congelador para 4 – 8 h e depois limpar, removendo o papel, enxaguando Ivo dos lados e andar com água quente e esfregar todas as superfícies com detergente e cloro. Lave as gaiolas com água quente e deixe para secar.</li>
</ol>

<ol>
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Os autores não têm nada para divulgar.

Estável de moscas encontram-se desenvolvendo em uma ampla variedade de substratos na natureza e podem ser criadas em muitos tipos de mídia no laboratório. Nós usamos aparas de madeira e vermiculita como agentes de volume. Vermiculita funcionou bem, mas fez separação pupários de criação meio difícil e era caro (~$0.60/pan). Possivelmente por causa do rigor adicionado de separar pupários do meio, eclosão também foi menor com vermiculita, 57% vs. 75% de aparas de madeira. Aparas de Cottonwood foram comparáveis...

Estável de moscas, Stomoxys calcitrans (L.), são hematófagos moscas cujas mordidas dolorosas interrompem o comportamento de pastoreio de gado, causam dor e sofrimento para animais de companhia e interromper atividades recreativas humanas em todo o mundo. Imaturas estáveis moscas desenvolvem-se em fermentação vegetativa passa, muitas vezes contaminada com resíduos animais. Culturas e práticas agronômicas mudanças produziram graves surtos de moscas estáveis em resíduos de culturas, legumes em Austrália1, cana-de-açúcar no Brasil2e abacaxi em Costa Rica3. Embora apenas 14 estáveis moscas por animal são consideradas o limite económico4, foram feitas observações de mais de 2.000 moscas por animal durante recentes surtos5. Esses níveis de infestação reduzem a produtividade de anfitrião para perto de zero e podem causar mortalidade6. Como resultado de surtos associados agronomicamente, moscas estáveis recebendo renovado interesse e demanda por colônias de laboratório tem aumentado dramaticamente.
Quanto a todos os insetos holometamorphic, moscas estáveis obter todos os nutrientes necessários para o crescimento durante o estágio larval ou imaturo. Portanto, um importante componente de um sistema de criação é a dieta larval ou substrato. Foram observadas larvas de mosca estáveis desenvolvimento em uma ampla gama de substratos no campo7 e eles são dependentes da comunidade microbiana do substrato8,9. Substratos larvas naturais são compostos principalmente de decomposição ou fermentação vegetativas materiais frequentemente contaminados com resíduos nitrogenados.
Para a criação de laboratório, substratos larvas moscas estáveis são geralmente compostos por um material vegetativo e uma fonte de nitrogênio adicionado. Vários materiais foram usados para dietas larvas moscas estáveis. As dietas larvas primeiras imitaram substratos naturais e incluíam da fermentação, palha de aveia e de cavalo ou vaca estrume10,11. Fontes de carboidratos incluem trigo farelo12,13,14, alfafa refeição12,13,14 e uma formulação comercial desenvolvido por especialidades químicas Fabricantes Association (CSMA, 33% de farelo de trigo, farinha de alfafa de 27%, levedura de cerveja 40% grânulos)13,14,15,16. Fontes de nitrogênio incluem levedura suspensão12, farinha de peixe e de bicarbonato de amónio17. Volume de materiais inertes são frequentemente incluídos nas dietas, incluindo aveia cascos12, bagaço13, vermiculita16, lascas de madeira13,18 e peletizadas hulls do amendoim14.
Um objetivo principal do laboratório de criação é de produzir um produto que é como fisiologicamente semelhante ao "tipo selvagem" quanto possível, a fim de que experimentos de laboratório produzirá resultados refletindo de populações do campo. Isto requer que na criação e seleção ser minimizados para manter genético diversidade e recursos nutricionais ser comparáveis no campo. Objetivos secundários são para minimizar o trabalho e despesas. Um componente importante de minimizar despesas é o uso de componentes de dieta disponíveis localmente. A mosca de estábulo criação sistema apresentado foi desenvolvida para atender a estes objectivos.

<table><tbody><tr><td>Diamalt</td><td>Premier Malt Products, Inc., Saddle Brook, NJ</td><td>2540</td><td></td></tr><tr><td>CSMA Fly media</td><td>Purina Animal Nutrition, Arden Hills, MN</td><td>5S6Z</td><td></td></tr><tr><td>Thin Maxi Pad</td><td>The Tranzonic Co., Cleveland, OH, USA</td><td>5001M</td><td></td></tr><tr><td>Calf Manna</td><td>MannaPro, Chesterfield, MO, USA</td><td>Manna Pro</td><td></td></tr><tr><td>Ammonium Bicarbonate</td><td>Spectrum Chemical Manufacturing Corp, Gardena, CA</td><td>A1125</td><td></td></tr><tr><td>Wheat bran, Coarse</td><td>Siemer Milling Company, Teutopolis, IL</td><td></td><td></td></tr><tr><td>Wood shavings</td><td>Tractor Supply Company, Brentwood, TN</td><td>502770699</td><td></td></tr><tr><td>Fishmeal</td><td>Consumer Supply Distributing, North Sioux City, SD</td><td>F1550</td><td></td></tr><tr><td>Adult cages</td><td>All Aluminum Window Company, Lincoln, Ne</td><td>Custom</td><td>45 &amp;times; 45 &amp;times; 45 cm, 18 &amp;times; 16 mesh aluminum screen, stockinette access</td></tr><tr><td>9 &amp;times; 28 cm black cotton cloth</td><td>Robert Kaufman Fabrics, Los Angeles, CA</td><td>K040-114</td><td>Egging cloth</td></tr><tr><td>10 L plastic dish pans</td><td>Rubbermaid, Saratoga Springs, NY</td><td>FG2951ARWHT</td><td>Larval pans</td></tr><tr><td>Stockinette, Cotton, 12 inch x 25 yard roll</td><td>Tex-Care Medical Company, Burlington, NC</td><td>91311-225</td><td></td></tr></tbody></table>

laboratory rearing of stable flies and other muscoid diptera

Estável de moscas, Stomoxys calcitrans, são pragas sérias de gado, seres humanos, animais de companhia e animais selvagens em todo o mundo. Durante os últimos 20 + anos, mudanças nas práticas agronômicas resultaram em graves surtos de moscas estáveis em vários países. Estes focos interrompeu a produção animal e recreação humana resultando em demandas públicas para aumentar os esforços de pesquisa e de gestão para esta praga. É apresentado um procedimento simples e de baixo custo para a criação de moscas estáveis para estudos de laboratório. O procedimento utiliza suprimentos, equipamentos e componentes da dieta disponíveis localmente. O procedimento pode ser adaptado para a criação de outras moscas de muscoid incluindo a mosca doméstica (Musca domestica), mosca do chifre (Haematobia irritans) e mosca do rosto (Musca autumnalis). O procedimento produz pupários mosca estável, com média de 12,5 mg e ~ 35% ovo para a sobrevivência de adulto. Cerca de 3000 moscas são produzidas em cada panela.

As larvas pupariate 10 a 14 dias e adultos emergem 14 – 16 dias após a oviposição. Tempo de geração, ovo a ovo, é ~ 24 dias. Dados de criação para maio de 2013 para de 2017 de Janeiro com três diferentes agentes de volume e duas colónias são apresentados na Figura 5. Cottonwood deu o melhor rendimento, 3867 ±1442 (<img alt="Equation" src="/files/ftp_upload/57341/57341eq1.jpg"> </img> ± desvio padrão) pupas pesando 12,5 mg ±1.6 com 74 ±19% ec...

Watch this Scientific Journal Video about Laboratory Rearing of Stable Flies and Other Muscoid Diptera at JoVE.com

Laboratory Rearing of Stable Flies and Other Muscoid Diptera

Um procedimento para a criação de moscas estáveis (Stomoxys calcitrans) é apresentado. O procedimento usa materiais localmente disponíveis para suprimentos, equipamentos e componentes de dieta.

Laboratório de criação de moscas estáveis e outros Muscoid Diptera

Stable flies, Stomoxys calcitrans, are serious pests of livestock, humans, companion animals and wildlife worldwide. During the last 20+ years, changes in ...

laboratory-rearing-of-stable-flies-and-other-muscoid-diptera

Research

JoVE Journal

Biology

8.5K visualizações.  ARS-USDA. Um procedimento para a criação de moscas estáveis (Stomoxys calcitrans) é apresentado. O procedimento usa materiais localmente disponíveis para suprimentos, equipamentos e componentes de dieta.

Vídeo: Laboratory Rearing of Stable Flies and Other Muscoid Diptera

Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions

The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is a model for understanding microbial interactions with animal hosts, facilitated by approaches to rear large sample sizes of Drosophila under microorganism-free (axenic) conditions, or with defined microbial communities (gnotobiotic). This work outlines a method for collection of Drosophila embryos, hypochlorite dechorionation and sterilization, and transfer to sterile diet. Sterilized embryos are transferred to sterile diet in 50 ml centrifuge tubes, and developing larvae and adults remain free of any exogenous microbes until the vials are opened. Alternatively, flies with a defined microbiota can be reared by inoculating sterile diet and embryos with microbial species of interest. We describe the introduction of 4 bacterial species to establish a representative gnotobiotic microbiota in Drosophila. Finally, we describe approaches for confirming bacterial community composition, including testing if axenic Drosophila remain bacteria-free into adulthood.

Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions

A method for rearing Drosophila melanogaster under axenic and gnotobiotic conditions is presented. Fly embryos are dechorionated in sodium hypochlorite, transferred aseptically to sterile diet, and reared in closed containers. Inoculating diet and embryos with bacteria leads to gnotobiotic associations, and bacterial presence is confirmed by plating whole-body Drosophila homogenates.

Elevando a mosca da fruta<em&gt; Drosophila melanogaster</em&gt; Sob axênicas e gnotobióticos Condições

The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is ...

Biologia do Desenvolvimento

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are also a useful scientific model to study motion vision and flight dynamics in a controlled laboratory setting. As the larvae develop in organically polluted water, they are useful models for investigating investment in microbial immunity. While large scale commercial breeding for agriculture already occurs, there are no standardized protocols for maintaining captive populations for scientific studies. This is important as commercial captive breeding programs focusing on mass output during peak pollination periods may fail to provide a population that is consistent, stable and robust throughout the year, as is often needed for other research purposes. Therefore, a method to establish, maintain and refresh a captive research population is required. Here, we describe the utilization of an artificial hibernation cycle, in addition to the nutritional and housing requirements, for long term maintenance of Eristalis tenax. Using these methods, we have significantly increased the health and longevity of captive populations of E. tenax compared to previous reports. We furthermore discuss small scale rearing methods and options for optimizing yields and manipulating population demographics.

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

The overall goal of these procedures is to establish, maintain and refresh a captive population of Eristalis tenax in a research setting.

Criação e manutenção de longo prazo de Eristalis tenax Hoverflies para estudos de investigação

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are ...

Ambiente

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

Um método para testar o efeito de sugestões ambientais no comportamento de acoplamento em<em&gt; Drosophila melanogaster</em

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

Comportamento

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are delivered using a non-quantitative method. Accurate delivery of known quantities of nucleic acids or other chemicals is critical to measure the efficacy of the compound being tested and to allow reliable comparison between compounds.
Here we provide a reproducible, quantitative microinjection protocol that ensures accurate delivery of specific doses of dsRNA, reducing the mortality typically induced by injection injury. These modifications include the addition of Rhodamine B, a graduated injection needle, and an improved recovery method borrowed from Isoe and Collins. This method allows calculation of dose responses and facilitates comparisons between compounds. Versions of this method have been successfully used on three genera of mosquitoes as well as house flies to assess the reduction in fecundity resulting from gene silencing of ribosomal RNA transcripts.
This protocol provides strategies to reduce several challenges of small insect microinjection. Together, mechanical delivery of dsRNAs accompanied by visual verification, identification of effective locations for delivery, and inclusion of a post-injection recovery period ensure accurate dosing and low injury mortality. This protocol also describes an oviposition bioassay for uniform determination of effects on fecundity.

This protocol describes a microinjection methodology that we have standardized and used for several years to deliver specific quantities of nucleic acids directly to the hemolymph of mosquitoes and house flies. This protocol results in minimal injection mortality and allows dose correlated measurements of fecundity.

Reprodutível dsRNA Microinjection e Bioassay de oviposição em mosquitos e moscas domésticas

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are ...

Flying Insect Detection and Classification with Inexpensive Sensors

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow for the development of many useful applications in vector and pest control for both medical and agricultural entomology. Given this, the last sixty years have seen many research efforts devoted to this task. To date, however, none of this research has had a lasting impact. In this work, we show that pseudo-acoustic optical sensors can produce superior data; that additional features, both intrinsic and extrinsic to the insect&#8217;s flight behavior, can be exploited to improve insect classification; that a Bayesian classification approach allows to efficiently learn classification models that are very robust to over-fitting, and a general classification framework allows to easily incorporate arbitrary number of features. We demonstrate the findings with large-scale experiments that dwarf all previous works combined, as measured by the number of insects and the number of species considered.

We proposed a system that uses inexpensive, noninvasive pseudo-acoustic optical sensors to automatically and accurately detect, count, and classify flying insects based on their flying sound.

Vôo Detecção e Classificação Insect com sensores baratos

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow ...

Bioengenharia

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

Fopius arisanus (Sonan) is an important parasitoid of Tephritid fruit flies for at least two reasons. First, it is the one of only three opiine parasitoids known to infect the host during the egg stage1. Second, it has a wide range of potential fruit fly hosts. Perhaps due to its life history, F. arisanus has been a successfully used for biological control of fruit flies in multiple tropical regions2-4. One impediment to the wide use of F. arisanus for fruit fly control is that it is difficult to establish a stable laboratory colony5-9. Despite this difficulty, in the 1990s USDA researchers developed a reliable method to maintain laboratory populations of F. arisanus10-12. There is significant interest in F. arisanus biology13,14, especially regarding its ability to colonize a wide variety of Tephritid hosts14-17; interest is especially driven by the alarming spread of Bactrocera fruit fly pests to new continents in the last decade18. Further research on F. arisanus and additional deployments of this species as a biological control agent will benefit from optimizations and improvements of rearing methods. In this protocol and associated video article we describe an optimized method for rearing F. arisanus based on a previously described approach12. The method we describe here allows rearing of F. arisanus in a small scale without the use of fruit, using materials available in tropical regions around the world and with relatively low manual labor requirements.

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

Fopius arisanus is an egg-larval parasitoid of Tephritid fruit flies that is successfully used in biological control of these important tropical pests. We describe here an optimized protocol for rearing F. arisanus on a small scale using readily available materials.

Um protocolo Optimizado para Criação Fopius arisanus, Um parasitóide de Frutas Tephritid Flies

Fopius arisanus (Sonan) is an important parasitoid of Tephritid fruit flies for at least two reasons. First, it is the one of only three opiine ...

Biologia

Maintenance of a Drosophila melanogaster Population Cage

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Maintenance of a Drosophila melanogaster Population Cage

This manuscript reports a detailed protocol for culturing, on a regular basis, a population of Drosophila melanogaster using a fly population cage.

Manutenção de um<em&gt; Drosophila melanogaster</em&gt; Gaiola População

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life ...

Circadian Entrainment of Drosophila Melanogaster

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this rhythm and to understand the resulting outputs, entrainment of model organisms to defined circadian time-points is required. Here we detail a procedure to entrain many Drosophila to a defined circadian rhythm. Furthermore, we detail post-entrainment steps to prepare samples for immunofluorescence, nucleic acid, or protein extraction-based analysis.

Here, we detail how to synchronize Drosophila to a circadian day. This is the first, and most important step necessary for studying biological rhythms and chronobiology.

Circadiano De Drosophila Melanogaster

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this ...

Topical Application Bioassay to Quantify Insecticide Toxicity for Mosquitoes and Fruit Flies

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. Insecticide resistance surveillance of mosquito populations is typically done through Centers for Disease Control and Prevention (CDC) bottle bioassays or World Health Organization (WHO) tube tests. However, these methods can result in a high degree of variability in mortality data due to variable insecticide contact with&#160;the insect, the relatively small numbers of organisms tested, extensive variation in mass between populations, and constantly changing environmental conditions, leading to variable outcomes. This paper presents the topical application bioassay, adapted as a high-throughput phenotypic bioassay for both mosquitoes and fruit flies, to test large numbers of insects along&#160;a range of insecticide concentrations.
This assay 1) ensures consistent treatment and insecticide contact with every organism, 2) produces highly specific dose-response curves that account for differences in average mass between strains and sexes (which is particularly important for field-collected organisms), and 3) allows for the calculation of statistically rigorous median lethal doses (LD50), which are necessary for resistance ratio comparisons-an alternative surveillance approach from diagnostic dose mortality, which is also used for larvicide resistance surveillance. This assay will be a complementary tool for accurately phenotyping mosquito populations and, as illustrated using fruit flies, is easily adaptable for use with other insects. We argue that this assay will help fill the gap between genotypic and phenotypic insecticide resistance in multiple insect species.

We describe the methodology and importance of the topical application bioassay to measure insecticide susceptibility in mosquitoes and fruit flies. The presented assay is high-throughput, utilizes insect mass-thus allowing for calculating a mass-relativized lethal dose instead of concentration-and likely has lower variability than other similar methods.

Bioensaio de aplicação tópico para quantificar toxicidade de inseticida para mosquitos e moscas de frutas

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. ...

Bradysia coprophila Lab Protocols and Chromosomal Research

Laboratory Maintenance of the Lower Dipteran Fly Bradysia (Sciara) coprophila: A New/Old Emerging Model Organism

Laboratory stocks of the lower dipteran fly, Bradysia (Sciara) coprophila, have been maintained for over a century. Protocols for laboratory upkeep of B. coprophila are presented here. These protocols will be useful for the rapidly increasing number of laboratories studying B. coprophila to take advantage of its unique biological features, which include (1) a monopolar spindle in male meiosis I; (2) non-disjunction of the X dyad in male meiosis II; (3) chromosome imprinting to distinguish maternal from paternal homologs; (4) germ line-limited (L) chromosomes; (5) chromosome elimination (paternal chromosomes in male meiosis I; one to two X chromosomes in early embryos; L chromosomes from the soma in early embryos); (6) sex determination by the mother (there is no Y chromosome); and (7) developmentally regulated DNA amplification at the DNA puff loci in larval salivary gland polytene chromosomes. 
It is now possible to explore these many unique features of chromosome mechanics by using the recent advances in sequencing and assembly of the B. coprophila genome and the development of transformation methodology for genomic engineering. The growing scientific community that uses B. coprophila for research will benefit from the protocols described here for mating the flies (phenotypic markers for mothers that will have only sons or only daughters; details of mass mating for biochemical experiments), checking embryo hatch, feeding larvae, and other comments on its rearing.

Author Spotlight: Exploring Bradysia coprophila's Unique Biology &#8211; A Guide to Laboratory Maintenance

This paper outlines the laboratory maintenance (including mating and feeding) of the lower dipteran fly Bradysia (Sciara) coprophila.

Manutenção laboratorial da Bradisia (Sciara) coprophila da mosca dos dípteros inferiores: um novo / antigo organismo modelo emergente

Laboratory stocks of the lower dipteran fly, Bradysia (Sciara) coprophila, have been maintained for over a century. Protocols for laboratory upkeep of B. ...

Laboratório de criação de moscas estáveis e outros Muscoid Diptera

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