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

Nos gustaría agradecer a Anthony Weinhold y los numerosos estudiantes que han trabajado con nosotros en los años de soporte técnico así como sugerencias para mejorar nuestro insecto procedimientos de crianza.

1. huevo colección (figura 1)
<ol><li>Preparar aplaudiéndote, coloque un extremo del paño en ~ 500 mL vaso de precipitados lleno de agua caliente (~ 40 ° C). Sobreponga los lados de la Copa y se fijan con una banda elástica. Doblar el extremo suelto de la tela sobre la parte superior de la Copa.</li>
	<li>Lugar Copa egging en jaula de moscas de 8 – 10 días para la hembras grávidas ~ 2 h. estable vuela se ovipositan en el paño.</li>
	<li>Retire la taza compañeros de jaula y enjuague los huevos fuera de la tela egging en una olla pequeña con botella de lavado del apretón.</li>
</ol>2. preparar Pan de cría Larval
<ol><li>
Preparar medio (cantidades para 1 bandeja de cría)
	<ol>
<li>Combinar el salvado de trigo (500 g), virutas de madera (200 g) y harina de pescado (115 g) de pan del plato de plástico de 10 L y mezclar bien.</li>
		<li>Añadir bicarbonato de amonio (50 g) agua (~ 25 ° C, 1600 mL) y mezcle hasta que se disuelva.</li>
		<li>Agregue una solución de agua a los ingredientes secos y mezclar para no unwetted material. Nivel medio en pan pero no comprimir.</li>
	</ol>
</li>
	<li>
Agregar los huevos de mosca estables
	<ol>
<li>Hacer un surco poco profundo en medio de la longitud de la bandeja.</li>
		<li>Depositar 1 mL de huevos de mosca estables (~ 8.000 huevos) en surco con pipeta de huevo (figura 2). Nota: Pipeta de huevo hecho cortando una pipeta graduada de plástico en el punto deseado, colocar 100 malla pantalla y pega pipeta volver junto.</li>
		<li>Cubra los huevos con una capa delgada del medio para evitar la desecación.</li>
		<li>Cubrir la cacerola con funda de almohada, cerrar con una banda de caucho y la etiqueta. Coloque bandejas en sala de larvas (23 ± 2 ° C, 30-50% HR y 12:12 [l:] h fotoperiodo). Nota: Desarrollo Larval toma 10-14 días desde el momento de la oviposición.</li>
	</ol>
</li>
</ol>3. procesamiento pupa, larvas hacia el borde del medio 7-9 días después Oviposition y Pupariate por días 13 o 14
<ol><li>Valores de Scoop, situada principalmente alrededor de los bordes de la bandeja debajo de la costra que forma sobre la superficie del medio, hacia fuera y coloca en una cacerola limpia.</li>
	<li>Llene la bandeja, con valores, ½ para ¾ de agua. Romper grumos de moscas y el medio. Componentes de salvado y proteína de trigo del medio se hunden mientras que valores > 1 día de viejo flotador19.</li>
	<li>Quitar materiales flotantes, moscas y algunos componentes del medio con un colador y lavar a través de una serie de tamices (#5, #7, #12, #20) para quitar el restante medio. El tamiz #12 recoge los valores y el #20 mantiene residuos entren en el desagüe. Valores entre tamices con un rociador Fregadero de lavado.</li>
	<li>Aclarar valores, en el tamiz #12, en un recipiente limpio. Llene la olla a ½ lleno de agua. Vierta moscas flotantes en un colador y deje que el agua sobrante al desagüe.</li>
	<li>Transferencia de escurrir moscas a pantalla de secado con un ventilador y dejar hasta que se seque (figura 3).</li>
</ol>4. estante pupa, un método alternativo para la recolección de moscas es con una pupa del estante18
Nota: El estante se hace de un trozo de plástico corte desde el extremo de una bandeja de plato 10 L (10,2 cm x 10,2 cm de ancho x 31,9 cm largo alto).
<ol><li>Preparar el medio como se describe en 2.1. Forma cónica medio pan de ~2.5 cm profundo en un extremo ~7.5 cm profundidad en el otro.</li>
	<li>
Coloque el estante en el medio en el extremo poco profundo del pan cría y cinta para evitar que las larvas arrastrándose junto a la plataforma (figura 4).
	<ol>
<li>Saturar la esponja (~14.5 x 9.0 x 4.5 cm3, celulosa regenerada) y envolverlo en un paño empapado de agua (30,5 x 42 cm2, algodón) y un lugar en el estante cerca de media pulgada desde el medio.</li>
	</ol>
</li>
	<li>
Agregue los huevos como se describe en 2.2 y proceder como antes.
	<ol>
<li>Asegurarse de cacerolas de todos los días que la esponja permanezca húmeda.</li>
	</ol>
</li>
	<li>Valores de enjuague de la plataforma, esponja y trapo en un plato limpio pan y vierta en el colador 14 días después de la oviposición.</li>
</ol>5. Control de calidad
<ol><li>Pesar todos los valores producidos en bandeja (peso total).</li>
	<li>Aislar 100 moscas, pesar y colocar en una placa de Petri de 9 cm.</li>
	<li>Cuenta y sexo adultos ~ 5-8 días más adelante para determinar la velocidad de emergencia y la sex ratio. Tienda platos de Petri con adultos emergidos en un congelador para contar en una fecha posterior si es necesario. Moscas son sexadas por la forma de sus ojos y el ancho de la placa fronto-orbital20 o genitales bajo magnificación baja.</li>
	<li>Registro de peso total, peso de 100 moscas y el número de emergidos machos y hembras.</li>
</ol>6. preparación de sangre
<ol><li>Recoger sangre bovina fresca de un matadero local en cubetas de 19 L que contiene 70 g de citrato de sodio dihidrato tribásico en 500 mL de agua. Remover vigorosamente por 5 a 10 min de la sangre y regresó al laboratorio.</li>
	<li>Sangre de tensión a través de un colador para quitar coágulos y repartido en los envases de 2 L, etiqueta con la fecha de colección y tienda en un congelador (-20 ° C).</li>
	<li>Retire el recipiente de lugar en refrigerador y congelador 2 días antes de la necesaria. Sangre puede utilizarse durante 2 semanas una vez descongeladas y almacenadas en el congelador hasta por 1 año.</li>
</ol>7. adulto mantenimiento
<ol><li>Preparación de jaulas, parte inferior de jaula de línea con papel de estraza para facilitar la limpieza.</li>
	<li>Colocar 50 g de pupas de mosca estables seco (~ 3.500) en cada jaula. Adultos eclose unas horas a un par de días después de la transformación pupal.</li>
	<li>Lugar fresco de la sangre había empapada femenino servilleta encima de cada jaula para la alimentación. Iniciar la alimentación dentro de las 24 h de eclosión adulto y repetir diariamente hasta un día antes de la oviposición (9 – 10 días después de la emergencia).</li>
	<li>Después de la oviposición (sección 1), coloque la jaula en el congelador durante 4-8 h y a continuación limpie quitando el papel, aclara Patricio desde los lados y el piso con agua caliente y restregar todas las superficies con detergente y lejía. Enjuagar las jaulas con agua caliente y deje que se seque.</li>
</ol>

<ol>
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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+for+collecting+clean+stable+fly+(Diptera:Muscidae)+pupae+of+known+age.">A new method for collecting clean stable fly (Diptera:Muscidae) pupae of known age.</a> Southwest. Entomol. 34 (4), 469-476 (2009).</li><li>Champlain, R. A., Fisk, F. W., Dowdy, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Some+improvements+in+rearing+stable+flies.">Some improvements in rearing stable flies.</a> J. Econ. Entomol. 47 (5), 940-941 (1954).</li><li>Zumpt, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The stomoxyine biting flies of the world. , (1973).</li><li>Wienhold, B. J., Taylor, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Substrate+properties+of+stable+fly+(Diptera:+Muscidae)+developmental+sites+associated+with+round+bale+hay+feeding+sites+in+eastern+Nebraska.">Substrate properties of stable fly (Diptera: Muscidae) developmental sites associated with round bale hay feeding sites in eastern Nebraska.</a> Environ. Entomol. 41 (2), 213-221 (2012).</li><li>Friesen, K., Berkebile, D. R., Wienhold, B. J., Durso, L., Zhu, J., Taylor, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+parameters+associated+with+stable+fly+(Diptera:+Muscidae)+development+at+hay+feeding+sites.">Environmental parameters associated with stable fly (Diptera: Muscidae) development at hay feeding sites.</a> Environ. Entomol. 45 (3), 570-576 (2016).</li><li>Albuquerque, T. A., Zurek, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+changes+in+the+bacterial+community+of+animal+feces+and+their+correlation+with+stable+fly+oviposition,+larval+development,+and+adult+fitness.">Temporal changes in the bacterial community of animal feces and their correlation with stable fly oviposition, larval development, and adult fitness.</a> Front. Microbiol. 5 (590), 1-9 (2014).</li><li>Bailey, D. L., Whitfield, T. L., LaBrecque, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+biology+and+techniques+for+mass+producing+the+stable+fly,+Stomoxys+calcitrans+(L.)+(Diptera:+Muscidae).">Laboratory biology and techniques for mass producing the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae).</a> J. Med. Entomol. 12 (2), 189-193 (1975).</li><li>Smith, J. 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Los autores no tienen nada que revelar.

Moscas de los establos se encuentran en una amplia variedad de substratos en la naturaleza y pueden criarse en muchos tipos de medios de comunicación en el laboratorio. Se han utilizado virutas de madera y vermiculita como agentes de carga. La vermiculita funcionó bien, pero separar valores de cría medio difícil y caro (~$0.60/pan). Posiblemente por el mayor rigor de separar los valores del medio, eclosión también fue menor con vermiculita, 57% vs. 75% de virutas de madera. Virutas de álamo eran comparables a la v...

Moscas de los establos, Stomoxys calcitrans (L.), son moscas hematófagas cuyas dolorosas picaduras alteran el comportamiento de pastoreo de ganado, causan dolor y sufrimiento a los animales del compañero e interrumpir actividades recreativas humanas en todo el mundo. Moscas de los establos se convierten en la fermentación de materia vegetal, a menudo contaminada con desechos animales. Cultivos y prácticas agronómicas cambiantes han producido graves brotes de mosca del establo en residuos de cosecha, hortalizas en Australia1, caña de azúcar en Brasil2y la piña en Costa Rica3. Aunque solo 14 estables moscas por animal se consideran el umbral económico4, se hicieron observaciones de más de 2.000 moscas por animal durante brotes recientes5. Esos niveles de infestación reducen la productividad de host para cerca de cero y pueden causar mortalidad6. Como resultado de agronómicamente asociados brotes, mosca del establo está recibiendo renovado interés y demanda de las colonias de laboratorio ha aumentado dramáticamente.
En cuanto a todos los holometamorphic insectos, moscas de los establos obtención todos los nutrientes requeridos para el crecimiento durante la etapa larval o inmaduro. Por lo tanto, un componente importante de un sistema de cría es la dieta larvaria o sustrato. Se han observado larvas de mosca estables en una amplia gama de substratos en el campo7 y son dependientes de la comunidad microbiana del sustrato8,9. Sustratos naturales de larvas se componen principalmente de descomposición o fermentación de materiales vegetativos a menudo contaminados con desechos nitrogenados.
Para la cría de laboratorio, sustratos larvas mosca estables se componen generalmente de un material vegetativo y una fuente de nitrógeno agregado. Numerosos materiales se han utilizado para las dietas de larvas mosca estables. La primera dieta larval mímico substratos naturales e incluye fermentación de paja de la avena y de10,de estiércol de caballo o vaca11. Fuentes de carbohidratos incluyen trigo salvado12,13,14, alfalfa comida12,13,14 y una formulación comercial desarrollado por especialidades químicas Fabricantes Asociación (CSMA, salvado de trigo 33%, 27% harina de alfalfa, levadura de cerveza de 40% gránulos)13,14,15,16. Fuentes de nitrógeno incluyen levaduras suspensión12, harina y bicarbonato de amonio17. Materias de carga inertes se incluyen a menudo en dietas incluyendo avena cascos12, bagazo13, vermiculita16, virutas de madera13,18 y cáscaras de maní pildoradas14.
Un objetivo primario de la cría de laboratorio es un producto que es como fisiológicamente similar a "wild type" como sea posible a fin de que experimentos de laboratorio producen resultados que reflejan los de poblaciones de campo. Esto requiere que en la cría y selección se reducen al mínimo para mantener la genética diversidad y recursos nutricionales ser comparables a ésos en el campo. Los objetivos secundarios son minimizar gastos y mano de obra. Un componente importante de minimizar los gastos es el uso de componentes de la dieta disponibles localmente. La mosca de los establos presentada sistema de crianza fue desarrollada para cumplir con estos objetivos.

<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

Moscas de los establos, Stomoxys calcitrans, son parásitos serios de la ganadería, los seres humanos, animales y vida silvestre en todo el mundo. Durante los últimos 20 años, cambios en las prácticas agronómicas dio lugar a graves brotes de mosca del establo en varios países. Estos brotes interrumpen la producción pecuaria y recreación humana, dando por resultado demandas del público para aumentar los esfuerzos de investigación y manejo de esta plaga. Se presenta un procedimiento simple y barato para la cría de moscas de los establos para los estudios de laboratorio. El procedimiento utiliza insumos, equipos y componentes de la dieta disponibles localmente. El procedimiento puede ser adaptado para la cría de otras moscas afines como mosca de la cara (Musca autumnalis), mosca de los cuernos (Haematobia irritans), mosca doméstica (Musca domestica). El procedimiento produce Moscas mosca estable con un promedio de 12,5 mg y ~ 35% de huevo para la supervivencia de adultos. Aproximadamente 3000 moscas se producen en cada pan.

Larvas pupariate 10 – 14 días y los adultos emergen de 14 a 16 días después de la oviposición. Tiempo de generación, huevo a huevo, es ~ 24 días. Datos de cría para mayo de 2013 a de 2017 de enero con tres diferentes agentes aumentadores de volumen y dos colonias se presentan en la figura 5. Cottonwood dio el mejor rendimiento, 3867 ±1442 (<img alt="Equation" src="/files/ftp_upload/57341/57341eq1.jpg"> </img> ± desviación estándar) pesa 12,5 mg 1...

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

Se presenta un procedimiento para la cría de moscas de los establos (Stomoxys calcitrans). El procedimiento utiliza materiales localmente disponibles para equipos, insumos y componentes de la dieta.

Laboratorio de cría de moscas de los establos y otros dípteros afines

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

8.5K vistas.  ARS-USDA. Se presenta un procedimiento para la cría de moscas de los establos (Stomoxys calcitrans). El procedimiento utiliza materiales localmente disponibles para equipos, insumos y componentes de la dieta.

Video: Laboratory Rearing of Stable Flies and Other Muscoid Diptera

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.

Cría y el mantenimiento a largo plazo de Eristalis tenax Tamaki para estudios de investigación

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

Investigación

Medio ambiente

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.

DsRNA reproducible microinyección y bioensayo de oviposición de mosquitos y moscas de la casa

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.

Flying detección de insectos y Clasificación con Sensores Barato

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

Bioingeniería

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.

Un protocolo optimizado para cría Fopius arisanus, Un parasitoide de moscas de fruta tefrítidas

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 ...

Biología

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.

El mantenimiento de una<em&gt; Drosophila melanogaster</em&gt; Jaula Población

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.

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

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human food. Outside its native range, culturing G. bimaculatus is feasible due to its dietary plasticity, rapid reproductive cycle, lack of diapause requirement, tolerance for high-density rearing, and robustness against pathogens. Thus, G. bimaculatus can be&#160;a versatile model for studies of insect physiology, behavior, embryology, or genetics.
Cultural parameters, such as stocking density, within-cage refugia, photoperiod, temperature, relative humidity, and diet, all impact cricket growth, behavior, and gene expression and should be standardized. In the burgeoning literature on farming insects for human consumption, these crickets are frequently employed to evaluate candidate feed admixtures derived from crop residues, food-processing byproducts, and other low-cost waste streams.
To support ongoing experiments evaluating G. bimaculatus growth performance and nutritional quality in response to variable feed substrates, a comprehensive set of standard protocols for breeding, upkeep, handling, measurement, and euthanasia in the laboratory was developed and is presented here. An industry-standard cricket feed has proven nutritionally adequate and functionally appropriate for the long-term maintenance of cricket breeding stocks, as well as for use as an experimental control feed. Rearing these crickets at a density of 0.005 crickets/cm3 in screen-topped 29.3 L polyethylene cages at an average temperature of 27 &#176;C on a 12 light (L)/12 dark (D) photoperiod, with moistened coconut coir serving both as hydration source and oviposition medium has successfully sustained healthy crickets over a 2-year span. Following these methods, crickets in a controlled experiment yielded an average mass of 0.724 g 0.190 g at harvest, with 89% survivorship and 68.2% sexual maturation between stocking (22 days) and harvest (65 days).

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

This paper outlines basic methods to standardize important factors such as density, feed availability, hydration source, and environmental controls for the long-term rearing of laboratory cultures of the edible cricket, Gryllus bimaculatus.

Mantenimiento de cultivos de laboratorio de Gryllus bimaculatus, un modelo versátil de ortópteros para la agricultura de insectos y la fisiología de invertebrados

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human ...

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.

Bioensayo de aplicación tópica para cuantificar la toxicidad de insecticidas para mosquitos y moscas de la fruta

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.

Mantenimiento de laboratorio de la mosca díptera inferior Bradysia (Sciara) coprophila: un nuevo/viejo 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. ...

Drosophila Maintenance

Drosophila melanogaster, commonly known as fruit flies, are a frequently used model organism for life science research. Although starting a collection of these critters may seem as easy as leaving a banana on your kitchen counter for too long, a productive fly colony in the lab requires careful husbandry and maintenance.
This video demonstrates the necessary steps for maintaining a healthy fly stock. The overview begins with the preparation and storage of the yeast and sugar-containing media on which flies feed. Next, the vessels most commonly used for housing Drosophila are shown, as well as how and when to move flies between these containers. Finally, the presentation also includes examples of the ways in which housing and feeding conditions are manipulated for biological experiments.

Maintenance and Husbandry of Drosophila melanogaster

Mantenimiento de Drosophila

Drosophila melanogaster, commonly known as fruit flies, are a frequently used model organism for life science research. Although starting a collection of ...