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>
	<li>Cook, D. F., Dadour, I. R., Keals, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+fly,+house+fly+(Diptera:+Muscidae),+and+other+nuisance+fly+development+in+poultry+litter+associated+with+horticultural+crop+production.">Stable fly, house fly (Diptera: Muscidae), and other nuisance fly development in poultry litter associated with horticultural crop production.</a> J. Econ. Entomol. 92 (6), 1352-1357 (1999).</li><li>Dominghetti, T. F., de Barros, A. T., Soares, C. O., Can&#231;ado, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stomoxys+calcitrans+(Diptera:+Muscidae)+outbreaks:+current+situation+and+future+outlook+with+emphasis+on+Brazil.">Stomoxys calcitrans (Diptera: Muscidae) outbreaks: current situation and future outlook with emphasis on Brazil.</a> Rev. Bras. Parasitol. Vet. 24 (4), 387-395 (2015).</li><li>Sol&#243;rzano, J. -. A., Guilles, J., Bravo, O., Vargas, C., Gomez-Bonilla, Y., Bingham, G., 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=Biology+and+trapping+of+stable+flies+(Diptera:+Muscidae)+developing+in+pineapple+residues+(Ananas+comosus)+in+Costa+Rica.">Biology and trapping of stable flies (Diptera: Muscidae) developing in pineapple residues (Ananas comosus) in Costa Rica.</a> J. Insect Sci. 15 (1), 145 (2015).</li><li>Berry, I. L., Stage, D. A., Campbell, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Populations+and+economic+impacts+of+stable+flies+on+cattle.">Populations and economic impacts of stable flies on cattle.</a> Trans. Am. Soc. Agric. Eng. 26, 873-877 (1983).</li><li>Taylor, D. B., Vreysen, J., Hendrichs, R., Cardoso Pereira, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Area-wide+management+of+stable+flies.">Area-wide management of stable flies.</a> Area-wide management of insect pests. , (2017).</li><li>Bishopp, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+stable+fly+(Stomoxys+calcitrans+L.),+an+important+live+stock+pest.">The stable fly (Stomoxys calcitrans L.), an important live stock pest.</a> J.Econ. Entomol. 6 (1), 112-126 (1913).</li><li>Hogsette, J. A., Ruff, J. P., Jones, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+fly+biology+and+control+in+northwest+Florida.">Stable fly biology and control in northwest Florida.</a> J. Agric. Entomol. 4 (1), 1-11 (1987).</li><li>Lysyk, T., Kalischuk-Tymensen, L., Selinger, L., Lancaster, R., Wever, L., Cheng, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearing+stable+fly+larvae+(Diptera:+Muscidae)+on+an+egg+yolk+medium.">Rearing stable fly larvae (Diptera: Muscidae) on an egg yolk medium.</a> J. Med. Entomol. 38, 382-388 (1999).</li><li>Romero, A., Broce, 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=Role+of+bacteria+in+the+oviposition+behaviour+and+larval+development+of+stable+flies.">Role of bacteria in the oviposition behaviour and larval development of stable flies.</a> Med. Vet. Entomol. 20, 115-121 (2006).</li><li>Glaser, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearing+flies+for+experimental+purposes+with+biological+notes.">Rearing flies for experimental purposes with biological notes.</a> J. Econ. Entomol. 17 (4), 486-496 (1924).</li><li>Melvin, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+studies+on+the+effect+of+flies+and+fly+sprays+on+cattle.">Physiological studies on the effect of flies and fly sprays on cattle.</a> J. Econ. Entomol. 25 (6), 1151-1164 (1932).</li><li>Doty, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Convenient+method+of+rearing+the+stable+fly.">Convenient method of rearing the stable fly.</a> J. Econ. Entomol. 30 (2), 367-369 (1937).</li><li>Bridges, A. C., Spates, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Larval+medium+for+the+stable+fly+Stomoxys+calcitrans+(L.).">Larval medium for the stable fly Stomoxys calcitrans (L.).</a> Southwest. Entomol. 8 (1), 6-10 (1983).</li><li>Hogsette, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+diets+for+production+of+house+flies+and+stable+flies+(Diptera:+Muscidae)+in+the+laboratory.">New diets for production of house flies and stable flies (Diptera: Muscidae) in the laboratory.</a> J. Econ. Entomol. 85 (6), 2291-2294 (1992).</li><li>McGregor, W. S., Dreiss, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearing+stable+flies+in+the+laboratory.">Rearing stable flies in the laboratory.</a> J. Econ. Entomol. 48 (3), 327-328 (1955).</li><li>Goodhue, L. D., Cantrel, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+vermiculite+in+medium+for+stable+fly+larvae.">The use of vermiculite in medium for stable fly larvae.</a> J. Econ. Entomol. 51 (2), 250 (1958).</li><li>Friesen, K., Berkebile, D. R., Zhu, J. 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=Augmenting+laboratory+rearing+of+stable+fly+(Diptera:+Muscidae)+larvae+with+ammoniacal+salts.">Augmenting laboratory rearing of stable fly (Diptera: Muscidae) larvae with ammoniacal salts.</a> J. Insect Sci. 17 (1), 1-6 (2017).</li><li>Berkebile, D. R., Weinhold, A. P., 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=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. P., Hall, R. D., Thomas, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Field+studies+on+mortality+of+the+immature+stages+of+the+stable+fly+(Diptera:+Muscidae).">Field studies on mortality of the immature stages of the stable fly (Diptera: Muscidae).</a> Environ. Entomol. 14 (6), 881-890 (1985).</li></ol>

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 &#215; 45 &#215; 45 cm, 18 &#215; 16 mesh aluminum screen, stockinette access</td>
		</tr>
		<tr>
			<td>9 &#215; 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 liter plastic dish pans</td>
			<td>Rubbermaid, Saratoga Springs, NY</td>
			<td>FG2951ARWHT</td>
			<td>Larval pans</td>
		</tr>
		<tr>
			<td>Stockinette, Cotton, 12&#34; X 25 yd 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

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

Microbial Communities in Nature and Laboratory - Interview

Comunidades microbianas en la naturaleza y de laboratorio - Entrevista

Investigación

Biología

Protocol for Mosquito Rearing (A. gambiae)

This protocol describes mosquito rearing in the insectary. The insectary rooms are maintained at 28&deg;C and ~80% humidity, with a 12 hr. day/night cycle. For this procedure, you'll need mosquito cages, 10% sterile sucrose solution, paper towels, beaker, whatman filter paper, glass feeders, human blood and serum, water bath, parafilm, distilled water, clean plastic trays, mosquito food (described below), mosquito net to cover the trays, vacuum, and a collection chamber to collect adults.

Protocol for Mosquito Rearing A. gambiae

This video illustrates the general techniques used to rear Anopheles gambiae in the laboratory. The methods for caring for laboratory mosquitoes are demonstrated through all stages of the organism's life cycle from larvae to pupae to blood-feeding adults.

Protocolo para la cría de los mosquitos (A. gambiae)

This protocol describes mosquito rearing in the insectary. The insectary rooms are maintained at 28&deg;C and ~80% humidity, with a 12 hr. day/night ...

Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite isolated from each other, with largely independent histories and scientific communities. However, the interface between these usually disparate fields is the developmental programs underlying acquisition of functional electrical signaling properties and differentiation of functional chemical synapses during the final phases of neural circuit formation. This interface is a critically important area for investigation. In Drosophila, these phases of functional development occur during a period of &lt;8 hours (at 25&deg;C) during the last third of embryogenesis. This late developmental period was long considered intractable to investigation owing to the deposition of a tough, impermeable epidermal cuticle. A breakthrough advance was the application of water-polymerizing surgical glue that can be locally applied to the cuticle to enable controlled dissection of late-stage embryos. With a dorsal longitudinal incision, the embryo can be laid flat, exposing the ventral nerve cord and body wall musculature to experimental investigation. This system has been heavily used to isolate and characterize genetic mutants that impair embryonic synapse formation, and thus reveal the molecular mechanisms governing the specification and differentiation of synapse connections and functional synaptic signaling properties.

Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures

This technique exposes the Drosophila embryonic neuromusculature for immunohistochemistry or electrophysiological recording. It is useful for studying early events in neuromuscular development or performing electrophysiology in mutants that cannot hatch.

La recolección y preparación de Drosophila Los embriones de registro electrofisiológico y otros procedimientos

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite ...

DNA Stable-Isotope Probing (DNA-SIP)

DNA stable-isotope probing (DNA-SIP) is a powerful technique for identifying active microorganisms that assimilate particular carbon substrates and nutrients into cellular biomass. As such, this cultivation-independent technique has been an important methodology for assigning metabolic function to the diverse communities inhabiting a wide range of terrestrial and aquatic environments. Following the incubation of an environmental sample with stable-isotope labelled compounds, extracted nucleic acid is subjected to density gradient ultracentrifugation and subsequent gradient fractionation to separate nucleic acids of differing densities. Purification of DNA from cesium chloride retrieves labelled and unlabelled DNA for subsequent molecular characterization (e.g. fingerprinting, microarrays, clone libraries, metagenomics). This JoVE video protocol provides visual step-by-step explanations of the protocol for density gradient ultracentrifugation, gradient fractionation and recovery of labelled DNA. The protocol also includes sample SIP data and highlights important tips and cautions that must be considered to ensure a successful DNA-SIP analysis.

DNA Stable-Isotope Probing DNA-SIP

DNA stable-isotope probing is a cultivation-independent method to identify and characterize active communities of microorganisms that are capable of utilizing specific substrates. Assimilation of substrate enriched in heavy isotope leads to incorporation of labelled atoms into microbial biomass. Density gradient ultracentrifugation retrieves labelled DNA for downstream molecular analyses.

ADN de isótopos estables de palpado (ADN-SIP)

DNA stable-isotope probing (DNA-SIP) is a powerful technique for identifying active microorganisms that assimilate particular carbon substrates and ...

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 ...

Aseptic Laboratory Techniques: Plating Methods

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: 
 
&#9679; Perform plating procedures without contaminating media. 
&#9679; Isolate single bacterial colonies by the streak-plating method. 
&#9679; Use pour-plating and spread-plating methods to determine the concentration of bacteria. 
&#9679; Perform soft agar overlays when working with phage. 
&#9679; Transfer bacterial cells from one plate to another using the replica-plating procedure. 
&#9679; Given an experimental task, select the appropriate plating method.

When working with media and reagents used to culture microorganisms, aseptic technique must be practiced to ensure contamination is minimized. A variety of plating methods are routinely used to isolate, propagate, or enumerate bacteria and phage, all of which incorporate procedures that maintain the sterility of experimental materials.

Técnicas de laboratorio asépticas: Métodos de Revestimiento

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies ...

Simple Homemade Tools to Handle Fruit Flies&#8212;Drosophila melanogaster

The fruit fly, Drosophila melanogaster, is widely used both in biological research and biology education. Handling adult flies is common but difficult in practice, as adult flies fly. Demonstrated here is how to make some simple and cost-effective tools to address difficult issues in the handling of Drosophila. Holes in foam stoppers are made and pipette tips or funnels are inserted into the holes. Flies then move only in one direction into the pipette tip/funnel assemblage, allowing efficient control of the transfer of adult Drosophila into or out of a vial. Existing protocols have been modified for cool-anesthetizing flies by chilling in crushed ice and transferring them onto a cold, hard icepack surface. The icepack is covered with a piece of medical gauze that keeps immobilized flies from the condensed water when examined under a stereomicroscope. The flies are finally euthanized for counting and sorting or discarded by microwaving. A bottle-shaped cage has also been developed for collecting eggs, as well as a labor-saving device and accompanying protocol for cleaning glass culture vials.

Described here is the use of several homemade tools to transfer, chill, and kill adult Drosophila, as well as to clean glass culture vials and collect eggs. These tools are easy to make and are rather efficient in handling Drosophila.

Herramientas caseras simples para manejar moscas de la fruta—Drosophila melanogaster

The fruit fly, Drosophila melanogaster, is widely used both in biological research and biology education. Handling adult flies is common but difficult in ...

Dissection of the Endolymphatic Sac from Mice

The study of mutant mouse models of human hearing and balance disorders has unraveled many structural and functional changes which may contribute to the human phenotypes. Although important progress has been done in the understanding of the development and function of the neurosensory epithelia of the cochlea and vestibula, limited knowledge is available regarding the development, cellular composition, molecular pathways and functional characteristics of the endolymphatic sac. This is, in large part, due to the difficulty of visualizing and microdissecting this tissue, which is an epithelium comprised of only one cell layer. The study presented here describes an approach to access and microdissect the endolymphatic sac from the wild-type mouse inner ear at different ages. The result of a similar dissection is shown in a pendrin-deficient mouse model of enlargement of the vestibular aqueduct. A transgenic mouse with a fluorescent endolymphatic sac is presented. This reporter mouse can be used to readily visualize the endolymphatic sac with limited dissection and determine its size. It can also be used as an educational tool to teach how to dissect the endolymphatic sac. These dissection procedures should facilitate further characterization of this understudied part of the inner ear.

<meta charset="utf8"></head><body>Disección del saco endolinfático de ratones

This study describes how to dissect the endolymphatic sac from the inner ear of the mouse at different ages. The result of a similar dissection is shown in an Slc26a4-null mouse model of enlargement of the endolymphatic sac. A transgenic mouse with a fluorescent reporter expressed in the endolymphatic sac is presented as a model to readily visualize the endolymphatic sac, and educational tool.

Disección del saco endolinfático de ratones

The study of mutant mouse models of human hearing and balance disorders has unraveled many structural and functional changes which may contribute to the ...

Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Diverse bacterial species secrete ~20-300 nm extracellular vesicles (EVs), comprised of lipids, proteins, nucleic acids, glycans, and other molecules derived from the parental cells. EVs function as intra- and inter-species communication vectors while also contributing to the interaction between bacteria and host organisms in the context of infection and colonization. Given the multitude of functions attributed to EVs in health and disease, there is a growing interest in isolating EVs for in vitro and in vivo studies. It was hypothesized that the separation of EVs based on physical properties, namely size, would facilitate the isolation of vesicles from diverse bacterial cultures. 
The isolation workflow consists of centrifugation, filtration, ultrafiltration, and size-exclusion chromatography (SEC) for the isolation of EVs from bacterial cultures. A pump-driven tangential flow filtration (TFF) step was incorporated to enhance scalability, enabling the isolation of material from liters of starting cell culture. Escherichia coli was used as a model system expressing EV-associated nanoluciferase and non-EV-associated mCherry as reporter proteins. The nanoluciferase was targeted to the EVs by fusing its N-terminus with cytolysin A. Early chromatography fractions containing 20-100 nm EVs with associated cytolysin A - nanoLuc were distinct from the later fractions containing the free proteins. The presence of EV-associated nanoluciferase was confirmed by immunogold labeling and transmission electron microscopy. This EV isolation workflow is applicable to other human gut-associated gram-negative and gram-positive bacterial species. In conclusion, combining centrifugation, filtration, ultrafiltration/TFF, and SEC enables scalable isolation of EVs from diverse bacterial species. Employing a standardized isolation workflow will facilitate comparative studies of microbial EVs across species.

Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Bacteria secrete nanometer-sized extracellular vesicles (EVs) carrying bioactive biological molecules. EV research focuses on understanding their biogenesis, role in microbe-microbe and host-microbe interactions and disease, as well as their potential therapeutic applications. A workflow for scalable isolation of EVs from various bacteria is presented to facilitate standardization of EV research.

Aislamiento escalable y purificación de vesículas extracelulares de Escherichia coli y otras bacterias

Diverse bacterial species secrete ~20-300 nm extracellular vesicles (EVs), comprised of lipids, proteins, nucleic acids, glycans, and other molecules ...

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

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Comunidades microbianas en la naturaleza y de laboratorio - Entrevista

Protocolo para la cría de los mosquitos (A. gambiae)

La recolección y preparación de Drosophila Los embriones de registro electrofisiológico y otros procedimientos

ADN de isótopos estables de palpado (ADN-SIP)

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

Técnicas de laboratorio asépticas: Métodos de Revestimiento

Herramientas caseras simples para manejar moscas de la fruta—Drosophila melanogaster

Disección del saco endolinfático de ratones

Disección del saco endolinfático de ratones

Aislamiento escalable y purificación de vesículas extracelulares de Escherichia coli y otras bacterias