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

Vorremmo ringraziare Anthony Weinhold e i numerosi studenti che hanno lavorato con noi nel corso degli anni per il supporto tecnico, nonché suggerimenti per migliorare il nostro insetto procedure di allevamento.

1. uovo insieme (Figura 1)
<ol><li>Preparare tazza incita, inserire un'estremità del panno in Becher ~ 500 mL riempita con acqua calda (~ 40 ° C). Si sovrappongono i lati della tazza e fissare con un elastico. Piegare l'estremità libera del panno indietro sopra la parte superiore della Coppa.</li>
	<li>Posto Coppa incita in gabbia di mosche di 8 – 10 giorni vecchio per femmina Gravid h. ~ 2 stabile vola sarà oviposit sul panno.</li>
	<li>Allontana da uova in gabbia e risciacquare fuori il panno egging egging calice in un pentolino con bottiglia di lavaggio a pressione.</li>
</ol>2. preparare Pan allevamento larvale
<ol><li>
Preparare il supporto (quantità per 1 vaschetta di allevamento)
	<ol>
<li>Unire la crusca di frumento (500 g), trucioli di legno (200 g) e farina di pesce (115g) in vaschetta di plastica piatto di 10 L e mescolare bene.</li>
		<li>Aggiungere bicarbonato di ammonio (50 g) di acqua (~ 25 ° C, 1600 mL) e mescolare fino a completa dissoluzione.</li>
		<li>Aggiungere acqua soluzione gli ingredienti secchi e mescolare affinché nessun materiale unwetted. Livello medio in padella, ma non comprimere.</li>
	</ol>
</li>
	<li>
Aggiungere le uova volare stabile
	<ol>
<li>Fare un solco poco profondo in mezzo la lunghezza della padella.</li>
		<li>Depositare 1 mL di stabile volare uova (~ 8.000 uova) in solco con pipetta di uovo (Figura 2). Nota: Pipetta uovo fatta tagliando una pipetta graduata in plastica al punto desiderato, inserendo 100 mesh schermo e incollaggio pipetta tornare insieme.</li>
		<li>Coprire le uova con un sottile strato di mezzo per prevenire il disseccamento.</li>
		<li>Coprire la pentola con federa, chiudere con un elastico e l'etichetta. Mettere pentole in camera larvale (23 ± 2 ° C, 30 – 50% RH e 12:12 [L:D] h fotoperiodo). Nota: Lo sviluppo larvale dura 10 – 14 giorni dal momento della deposizione delle uova.</li>
	</ol>
</li>
</ol>3. pupal elaborazione, larve spostarsi sul margine del mezzo 7 – 9 giorni dopo la deposizione delle uova e Pupariate di giorni 13 o 14
<ol><li>Paletta puparia, principalmente situato intorno ai bordi della vaschetta sotto la crosta che si forma sulla superficie del mezzo, fuori e messo in una pentola pulita.</li>
	<li>Riempire la pentola, con puparia, ½ a ¾ piena d'acqua. Rompere con ciuffi di puparia e medie. Componenti di crusca e proteine di frumento del mezzo lavandino mentre puparia > 1 giorno vecchio galleggiante19.</li>
	<li>Rimuovere materiale galleggiante, puparia e alcuni componenti medi con un colino e lavare attraverso una serie di setacci (n. 5, #7, #12, #20) per rimuovere il mezzo rimanente. Il setaccio #12 raccoglie il puparia e il #20 mantiene rifiuti solidi di entrare lo scarico. Lavare puparia tra setacci con uno spruzzatore di lavandino.</li>
	<li>Risciacquare il setaccio #12, puparia, in una pentola pulita. Riempire la pentola a ½ pieno di acqua. Versare puparia galleggiante in un colino e lasciare drenare l'acqua in eccesso.</li>
	<li>Trasferimento drenati puparia a schermo di secchezza con ventilatore e lasciare fino a secco (Figura 3).</li>
</ol>4. pupal scaffale, un metodo alternativo per la raccolta Puparia è con un ripiano Pupal18
Nota: La mensola fatta da un pezzo di plastica tagliato dalla fine di una padella piatto tipico di 10 L (10,2 cm di altezza x 10,2 cm di larghezza x 31,9 cm di lunghezza).
<ol><li>Preparare il supporto come descritto al punto 2.1. Cono medio in padella da ~2.5 cm profondo su un'estremità a ~7.5 cm profondo su altro.</li>
	<li>
Inserire ripiano su supporto nella parte bassa di allevamento padella e nastro adesivo per evitare le larve dallo strisciare lungo la mensola (Figura 4).
	<ol>
<li>Saturare la spugna (~14.5 x 9.0 x 4,5 cm3, cellulosa rigenerata) e avvolgerlo in un panno imbevuto di acqua (30,5 x 42 cm2, cotone) e posto sullo scaffale circa mezzo pollice dal mezzo.</li>
	</ol>
</li>
	<li>
Aggiungere le uova, come descritto al punto 2.2 e procedere come prima.
	<ol>
<li>Verifica padelle ogni giorno per assicurare che la spugna rimanga umida.</li>
	</ol>
</li>
	<li>Risciacquo puparia dallo scaffale, spugna e panno in un piatto pulito padella e versare nel colino 14 giorni dopo la deposizione delle uova.</li>
</ol>5. controllo di qualità
<ol><li>Pesare tutti i puparia prodotta in padella (Totale wt.).</li>
	<li>Isolare 100 puparia, pesare e inseriti in una capsula di Petri di 9 cm.</li>
	<li>Conteggio e sesso adulti ~ 5 – 8 giorni più tardi per determinare il tasso di emersione e rapporto del sesso. Conservare capsule di Petri con adulti è emersi in un congelatore per il conteggio in un secondo momento, se necessario. Mosche sono sessuati dalla forma dei loro occhi e larghezza della placca fronto-orbitale20 o genitali sotto ingrandimento basso.</li>
	<li>Peso totale record, peso di 100 puparia e numero di emersi adulti maschi e femmine.</li>
</ol>6. preparazione di sangue
<ol><li>Raccogliere sangue fresco di bovini da un macello locale a secchi 19 L contenenti 70 g di sodio citrato tribasico diidrato in 500 mL di acqua. Mescolare vigorosamente per 5 – 10 min di sangue e restituito al laboratorio.</li>
	<li>Sangue di ceppo attraverso un colino per rimuovere la formazione di coaguli e partizionato in contenitori 2L, etichetta con la data di raccolta e conservare in congelatore (-20 ° C).</li>
	<li>Rimuovere il contenitore dal freezer e posto in frigorifero 2 giorni prima del necessario. Sangue può essere utilizzato per ~ 2 settimane una volta scongelati e conservati nel congelatore per fino a 1 anno.</li>
</ol>7. adult Maintenance
<ol><li>Preparare gabbie, fondo gabbia linea con carta da macellaio per facilitare la pulizia.</li>
	<li>Mettere 50 g di secco pupe volare stabile (~ 3.500) in ogni gabbia. Adulti eclose poche ore a un paio di giorni dopo l'elaborazione pupal.</li>
	<li>Posto un sangue fresco imbevuto femminile tovagliolo sulla cima di ogni gabbia per l'alimentazione. Avviare l'alimentazione entro 24 h dalla eclosion adulto e ripetere ogni giorno fino al giorno prima della deposizione delle uova (9 – 10 giorni dopo la nascita).</li>
	<li>Dopo la deposizione delle uova (sezione 1), posizionare la gabbia in freezer per 4 – 8 h e poi pulire rimuovendo la carta, risciacquo frass dai lati e piano con acqua calda e strofinare tutte le superfici con detersivo e candeggina. Sciacquare le gabbie con acqua calda e lasciare per asciugare.</li>
</ol>

<ol>
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Mosche stabili possono trovare lo sviluppo in un'ampia varietà di substrati in natura e possono essere allevati in molti tipi di media in laboratorio. Abbiamo usato dei trucioli di legno e vermiculite come agenti volumizzanti. La vermiculite ha funzionato bene, ma fatto puparia separazione da allevamento medio difficile ed era costoso (~$0.60/pan). Forse a causa il rigore aggiunto di separare puparia dal mezzo, eclosion era inoltre più basso con vermiculite, 57% vs 75% per trucioli di legno. Trucioli di Cottonwood eran...

Mosche stabili, Stomoxys calcitrans (L.), sono ematofagi mosche cui punture dolorose può interferire il comportamento al pascolo del bestiame, causano dolore e sofferenza per animali da compagnia e interferire con attività ricreative umane in tutto il mondo. Mosche stabili immaturi sviluppano nella fermentazione materia vegetativa, spesso contaminato da rifiuti di origine animale. Colture e mutevoli pratiche agronomiche hanno prodotto gravi episodi di mosche stabili in presenza di residui colturali, verdure in Australia1, canna da zucchero in Brasile2e l'ananas in Costa Rica3. Anche se solo 14 mosche stabili per animale sono considerati la soglia economica4, sono state fatte osservazioni di più di 2.000 mosche per animale durante recenti focolai5. Tali livelli di infestazione riducono la produttività di host per vicino allo zero e possono causare mortalità6. A seguito di focolai agronomico associati, mosche stabili stanno ricevendo rinnovato interesse e domanda per colonie di laboratorio è aumentato drammaticamente.
Per quanto riguarda tutti gli insetti holometamorphic, mosche stabili ottenere tutti i nutrienti necessari per la crescita durante la fase larvale o immatura. Pertanto, una componente importante di un sistema di allevamento è la dieta larvale o substrato. Le larve del Mosca stabile sono state osservate lo sviluppo in un'ampia gamma di substrati in campo7 e sono dipendente della comunità microbica del substrato8,9. Substrati naturali larvali sono principalmente composte di decomposizione o vegetativi materiali spesso contaminati con rifiuti azotati di fermentazione.
Per l'allevamento di laboratorio, substrati larvale volare stabile sono di solito composte da un materiale vegetativo e una fonte di azoto aggiunto. Numerosi materiali sono stati usati per le diete larvale volare stabile. Le diete larvale prime ha imitato substrati naturali e incluso di fermentazione paglia di avena e letame di cavallo o mucca10,11. Fonti di carboidrati includono frumento crusca12,13,14, erba medica pasto12,13,14 e una formulazione commerciale sviluppato da specialità chimiche Produttori Association (CSMA, crusca di grano 33%, 27% di farina di erba medica, lievito di birra 40% granuli)13,14,15,16. Fonti di azoto includono lievito sospensione12, farina di pesce e di bicarbonato di ammonio17. Materiali inerti volumizzanti sono spesso inclusi nelle diete tra cui avena scafi12, bagassa13, vermiculite16, trucioli di legno13,18 e gusci di arachidi pellettato14.
Un obiettivo primario di laboratorio allevamento è quello di produrre un prodotto che è come fisiologicamente simile a "wild type" possibile in modo che gli esperimenti del laboratorio produrrà risultati riflettono quelle delle popolazioni di campo. Ciò richiede che essere minimizzato in allevamento e selezione per mantenere genetica diversità e risorse nutritive essere paragonabili a quelle nel campo. Gli obiettivi secondari sono per ridurre al minimo il lavoro e le spese. Una componente importante di minimizzare le spese è l'utilizzo di componenti di alimentazione disponibile localmente. Al volo stabile del sistema presentato di allevamento è stato sviluppato per raggiungere questi obiettivi.

<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

Mosche stabili, Stomoxys calcitrans, sono parassiti gravi di bestiame, gli esseri umani, animali da compagnia e animali selvatici in tutto il mondo. Durante gli ultimi 20 + anni, cambiamenti nelle pratiche agronomiche ha provocato gravi epidemie di mosche stabili in diversi paesi. Questi focolai perturbato produzione zootecnica e ricreazione umana conseguente richieste pubbliche per aumentare gli sforzi di ricerca e gestione per questo parassita. Una procedura semplice e poco costoso per l'allevamento di mosche stabili per le ricerche di laboratorio è presentata. La procedura utilizza componenti localmente disponibili dieta, attrezzature e forniture. La procedura può essere adattata per l'allevamento di altre mosche muscoidi inclusi viso volare (Musca autumnalis), corno fly (da schistosoma haematobium irritans) e Mosca domestica (Musca domestica). La procedura produce puparia Vola stabile con una media di 12,5 mg e ~ 35% uovo alla sopravvivenza adulto. Circa 3000 mosche sono prodotte in ogni vaschetta.

Pupariate 10 – 14 giorni di larve e adulti emergono 14 – 16 giorni dopo la deposizione delle uova. Tempo di generazione, uovo, è ~ 24 giorni. Allevamento dei dati per maggio 2013 al gennaio 2017 con tre diversi agenti volumizzanti e due colonie sono presentati nella Figura 5. Cottonwood ha dato il miglior rendimento, 3867 ±1442 (<img alt="Equation" src="/files/ftp_upload/57341/57341eq1.jpg"> </img> ± deviazione standard) pupe peso 12,5 ± 1,6 mg con 74...

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

Una procedura per l'allevamento di mosche stabili (Stomoxys calcitrans) è presentata. La procedura utilizza materiali localmente disponibili per dieta componenti, attrezzature e forniture.

Laboratorio di allevamento di mosche stabili e altri ditteri muscoidi

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 Views.  ARS-USDA. Una procedura per l'allevamento di mosche stabili (Stomoxys calcitrans) è presentata. La procedura utilizza materiali localmente disponibili per dieta componenti, attrezzature e forniture.

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.

Allevamento e manutenzione a lungo termine di sirfidi Eristalis tenax per studi di ricerca

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

Ricerca

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.

Riproducibile dsRNA Microinjection e analisi biologica di deposizione delle uova di zanzare e mosche domestiche

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.

Volare Insetto rilevamento e classificazione con sensori Economico

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

Bioingegneria

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 protocollo ottimizzato per la cura dei Fopius arisanus, Un parassitoide di frutta Tephritid mosche

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.

Mantenimento di un<em&gt; Drosophila melanogaster</em&gt; Cage Popolazione

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.

Ingresso circadiano della Drosophila Melanogaster

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.

Mantenimento di colture di laboratorio di Gryllus bimaculatus, un modello ortottero versatile per l'agricoltura degli insetti e la fisiologia degli invertebrati

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.

Biotest di applicazione topica per quantificare la tossicità degli insetticidi per zanzare e moscerini della frutta

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.

Manutenzione in laboratorio della mosca dei ditteri inferiori Bradysia (Sciara) coprophila: un nuovo/vecchio organismo modello 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

Drosophila Manutenzione

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