We are grateful to anonymous reviewers for comments and suggestions that greatly improved this manuscript. We thank R. Bowes, R. Carreno, and T. Panhuis for assistance in collecting the fish. We also thank M. Lee for assistance with male preference trials. We are grateful to the Ohio Wesleyan University Department of Zoology for helpful advice and suggestions throughout this study and Arizona State University (McGraw Lab) for software advice.

Tutti gli esperimenti sono stati condotti con l&#39;approvazione del Comitato di Cura e uso istituzionale degli animali Ohio Wesleyan University. 1. Registrazione UV riflettanza di pesce utilizzando uno spettrofotometro <ol><li> Calibrare uno spettrofotometro e fonte di luce con uno standard bianco nota nella gamma di lunghezze d&#39;onda da misurare secondo le raccomandazioni dello strumento o software. </li><li> Anesthetize pesci mettendo in una soluzione allo 0,5% di etil 3-amminobenzoato metano sale di acido solfonico (MS-222) tamponato con un&#39;eguale quantità di bicarbonato di sodio fino pesci non risponde. Una volta non risponde, subito mettere il pesce su un nero, sfondo non riflettente. Nota: MS-222 è potenzialmente tossici, e guanti devono essere indossati in ogni momento. </li><li> Prendere tutte le misurazioni in una stanza buia per minimizzare luce in eccesso oltre quello della sorgente luminosa spettrometro. </li><li> riflettanza misura utilizzando una sonda a fibre ottiche tenuta contro il corpo dell&#39;organismo. Renderesicuri per formare un angolo di 45 ° coerente tra le fibre ottiche e il corpo, come è incorporato in alcuni disegni sonda. Nota: Per coerenza misura, può essere desiderabile media su più scansioni (2 - 5) per regione di body. </li><li> registrare i dati di riflettanza con adeguato software di analisi di riflettanza. Nota: il software utilizzato qui è stato impostato su: all&#39;integrazione tempo = 2.000 sec, scansioni media = 2, e pixel levigante = 5. I dati sono stati copiato e incollato ad un foglio di calcolo. </li><li> Ripetere su più zone del corpo per determinare la presenza o assenza di riflettanza UV e la gamma di variazione fenotipica di tutti gli organismi. Nota: possono essere necessarie scansioni preliminari per determinare quale, se del caso, le zone del corpo dimostrano costantemente le marcature UV. </li><li> Riportare il pesce per un acquario partecipazione ben ossigenata fino alla completa guarigione, come evidenziato da normali battiti opercolo e comportamenti di nuoto. Nota: Un protectant commerciale per ripristinare il rivestimento mucoso al corpo può essere aggiunto per aiutare nel recupero pelle. </li><li> rappresentantemangiare su più individui, tra cui entrambi i sessi, se desiderato. </li><li> Sul foglio di calcolo in cui tutti i dati sono stati copiati, i dati del diagramma utilizzando un grafico a dispersione, con riflessione sulla Y e lunghezza d&#39;onda sulla X-asse. picchi di ampiezza della gamma UV (300-400 nm) indicano la presenza di riflettanza UV sul pesce. </li></ol> 2. Prove di accoppiamento dicotomiche NOTA: effettuare osservazioni di controllo, in cui i pesci focale, filtri UV e filtri opachi sono in atto. In questi studi, quando vengono rimossi i filtri opachi, pesce focale sarà esposto a parti del serbatoio visualizzazione presenza e in assenza di luce UV, ma privi potenziali partner di accoppiamento. Per i dettagli sulla validità dei test a scelta dicotomici in questo esperimento vedere Palmer e Hankison (2015) 10, anche se nota che questi test hanno anche svantaggi 11. <ol><li> Utilizzare gli individui sessualmente ricettive come organismi focali. Utilizzare femminile P. latipinna &lt;48 ore dopo il parto 5 unnd maschi isolati per almeno 24 ore prima del test per migliorare la probabilità di motivazione sessuale. </li><li> Posizionare un singolo individuo focale in una zona neutra adiacente a due scomparti di scelta. Per P. latipinna, utilizzare un test acquario 75,7 L (30,5 cm x 30,5 cm x 75 cm; Figura 1) diviso in tre sezioni da setti plexiglas trasparenti, formando una zona centrale per l&#39;individuo focale affiancato da due compartimenti scelta di dimensioni uguali. Assicurarsi che l&#39;acqua o olfattivi segnali sono condivisi tra le singole sezioni del serbatoio. Ripetere per ogni individuo di essere osservato. </li></ol><img alt="Figura 1" src="/files/ftp_upload/54453/54453fig1.jpg" /> Figura 1. sperimentale Acquario installazione di un acquario rettangolare divisa in tre sezioni:. Un&#39;area centrale che ha tenuto i due compartimenti finali che hanno tenuto le coppie singolo oggetto focale e (la figura mostra una donna a metà con i maschi degli oggetti alle estremità). fil intercambiabileTERS che o riflettanza UV bloccato o consentito potrebbe essere collocato sopra i divisori in vetro che dividono le sezioni maschili e femminili dell&#39;acquario. Questa cifra viene modificato da Palmer e Hankison (2015) 10. <a href="http://ecsource.jove.com/files/ftp_upload/54453/54453fig1large.jpg" target="_blank">Si prega di cliccare qui per vedere una versione più grande di questa figura.</a> <ol start="3"><li> In condizioni di luce a spettro completo (illuminazione il cui spettro si estende nella gamma UV-A), isolare un potenziale partner di accoppiamento in ciascun compartimento fine (vano scelta) dell&#39;acquario di prova. Assicurarsi che potenziali partner di accoppiamento assomigliano più strettamente possibile in dimensioni, colorazione e visualizzazione caratteristiche, in modo che la differenza principale tra le due parti di accoppiamento isolati è se appaiono dietro un filtro che blocca o trasmette la luce UV. NOTA: Può essere preferibile registrare riflettanza nello spettro visibile per consentire la migliore corrispondenza dei colori possibile, alanche se qui abbinamento è stato fatto da occhio. </li><li> Ambientarsi individui sotto isolamento visivo (filtri opachi) per 15 min. Assicurarsi che UV + e - filtri sono in atto in questo momento. A seguito di acclimatazione, rimuovere i filtri opachi. Per garantire che le polarizzazioni collaterali non sono presenti (preferenza latente in un lato della vasca indipendentemente oggetto), assegnare casualmente l&#39;+ UV e UV - lati del serbatoio per ogni ogni prova. </li><li> Registrare la quantità di tempo che l&#39;individuo focale spende all&#39;interno delle zone di preferenza specifici (zone vicino i comparti di scelta, in questo caso di circa due lunghezze corpo di serie) utilizzando cronometri. Per pesce femmina, questa è la quantità di tempo che trascorre vicino ciascuno dei due maschi, e viceversa per il pesce focale maschio. NOTA: software del registratore evento può anche essere usato per la registrazione. Un quarto d&#39;ora le prove sono stati utilizzati in questo esperimento e pesce che non ha visitato ciascuna zona preferenza almeno una volta sono stati esclusi dai risultati come non risponde. </li><li> Registrare e analizzare il tempo che i pesci passano vicino a vuoto UV + e UV - scomparti per assicurare che non vi è una tendenza intrinseca verso un particolare ambiente di illuminazione che possono influenzare la scelta accoppiamento del pesce. Utilizzare appaiati t-test per determinare se gli individui preferiscono una particolare condizione UV. </li></ol>

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The authors have nothing to disclose.

Spettrofotometria è riuscito a identificare le marcature UV su P. latipinna. Entrambi i sessi di P. latipinna possiedono marcature UV lungo i loro lati. Inoltre, alcuni maschi avevano marcature UV sulle loro pinne dorsali, tratti precedentemente trovato per essere importante nelle preferenze di accoppiamento femmina 7. Si consiglia di utilizzare spettrofotometria UV come meccanismo per rilevare la presenza di marcature UV. Ulteriori test in grado di determinare ...

Capire gli indizi e segnali utilizzati nelle interazioni sociali animali ci permette di comprendere la variazione fenotipica sia all&#39;interno che tra le specie. Questa variazione gioca un ruolo importante nei processi evolutivi come divergenza popolazione, selezione sessuale, e speciazione. Spesso, tuttavia, i ricercatori sono limitati a esplorare i segnali più evidenti di sistemi sensoriali umani, in particolare quelli nei regni visivi o uditivi. L&#39;uso della spettrofotometria, però, ci permette di espandere le nostre indagini al di là dello spettro visibile umana e in lunghezze d&#39;onda che possono essere importanti nelle interazioni sociali in altre specie. In particolare, la comunicazione a corto raggio offerta da raggi ultravioletti (UV; 300-400 nm) sensibilità ha il potenziale per essere molto vantaggiosa durante scelta del compagno 1. Molti predatori visivamente-caccia di uccelli e pesci, per esempio, sono in grado di rilevare le radiazioni UV. Nei sistemi in cui i maschi mostrano elaboratoalle femmine, questi maschi potrebbero ridurre il rischio di predazione, pur mantenendo la loro capacità di attrarre i compagni sfruttando lo spettro UV, piuttosto che sviluppare spunti rilevabile nello spettro visibile 2,3 .. Se uno non riesce a prendere in considerazione la possibilità che gli organismi comunicano con ogni altri che utilizzano questi &quot;canali di comunicazione privati&quot;, i conducenti significative di comportamento ed evoluzione possono perdere. Questo protocollo descrive un&#39;indagine l&#39;uso di segnali UV per la scelta del compagno nel sailfin Molly, Poecilia latipinna, un pesce che ha poligama non precedentemente nota capacità di rilevare UV o utilizzare le marcature UV. Questa specie di pesci ha una stretta vicinanza filogenetica ad altri pesci live-cuscinetto sensibili ai raggi UV-4 e vi sono prove che microspectroscopic P. latipinna, insieme ad altre specie molly come P. mexicana e P. formosa, in possesso di una classe di coni (fotorecettori responsabili della covisione Lor) che sono più sensibili per lunghezze d&#39;onda UV 5. In questa specie dimorfismo sessuale, la scelta femminile ha svolto un ruolo forte per l&#39;evoluzione della brillantemente pinne colorati e ingranditi dei maschi 6-9. Questa metodologia ci permette di esplorare se UV è un mezzo ulteriore per cui le femmine valutare la qualità maschile. La rilevazione e la misurazione delle marcature UV su P. latipinna utilizzando uno spettrofotometro con una sonda a fibre ottiche è descritto qui. Inoltre, se ricettive Molly femminili differenziale associano con i maschi visti attraverso un filtro ottico che trasmettono la luce a spettro completo, tra cui UV-A ([UV +], 320-700 nm) e maschi visti attraverso un filtro UV-bloccante ([UV]; 400 - 700 nm) è discusso. Questo metodo ha ampie applicazioni per scoprire i modelli di sensibilità ai raggi UV e colore in pesci e altri organismi, consentendo la ricerca in una serie di domande che coinvolgono UV e il suo ruolo nel comportamento.

<table><tbody><tr><td>Spectrophotometer, P1000</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>DT 1000 xenon UV light source</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>Ocean Optics Overture Software</td><td>Ocean Optics</td><td></td><td>newer software is available</td></tr><tr><td>R200-Angle-UV bifurcated fiber-optic probe (Guided Wave)</td><td>Ocean Optics</td><td></td><td>newer models are availabe</td></tr><tr><td>Certified reflectance standard, white</td><td>labsphere</td><td></td><td></td></tr><tr><td>75.7 L Aquarium, divided</td><td>Experimental Builder</td><td></td><td></td></tr><tr><td>Full Spectrum Bulb</td><td>Nature&#039;s Sunlight</td><td></td><td></td></tr><tr><td>UV blocking sheet</td><td>GAM UV Sheet</td><td></td><td></td></tr></tbody></table>

assays to detect uv-reflecting structures and determine their importance in mate preference using the sailfin molly poecilia latipinna

Molti organismi utilizzano spunti e segnali al di là di sensibilità umana durante le interazioni sociali. E &#39;importante prendere in considerazione come gli organismi percepiscono i loro mondi quando si cerca di capire il loro comportamento e l&#39;ecologia. La sensibilità alla spettro ultravioletto (UV; 300-400 nm) si trova in tutta generi multiplo di uccelli, pesci, rettili, anfibi, e persino mammiferi. Questo protocollo descrive una tecnica per l&#39;esame organismi per la presenza di strutture UV-riflettente ed un metodo per verificare se questi segnali sono usati come segnali sociali nel contesto della scelta del compagno. Uno spettrofotometro è utilizzato per rilevare la presenza di riflettanza UV e variazione di intensità riflettente tra individui e sessi. Un esempio di questa tecnica è presentato in cui un test a scelta dicotomica compagno espone gli individui sessualmente ricettive per gli individui di sesso opposto il cui aspetto visivo può essere manipolato dai filtri che o trasmettono a spettro completo o il blocco UV lunghezze d&#39;onda. Questosistema ha consentito per la determinazione che femminili, ma non di sesso maschile, Molly Sailfin (Poecilia latipinna) stavano usando le marcature UV come parte delle loro decisioni di accoppiamento. Questi tipi di studi servono a espandere la nostra conoscenza della gamma di organismi che utilizzano raggi UV e fornire informazioni su come UV gioca un ruolo nella loro vita.

<html> Le figure 1 e 2 mostrano l&#39;accoppiamento preferenza istituito acquario e siti di misura UV per i nostri esperimenti. Misurare riflettanza UV ha permesso per la determinazione che P. latipinna possiedono caratteristiche UV, in particolare lungo i lati dei loro corpi (Figura 3), in aggiunta alla variazione individuale in questi tratti. Una volta tratti UV sono...

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Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

Questo protocollo descrive l&#39;uso di spettrofotometria di rilevare strutture ultravioletti che riflette sugli organismi (in questo esempio, il sailfin Molly Poecilia latipinna) e descrive test a scelta dicotomiche per i pesci che permettono inferenze da apportare sul ruolo di spunti ultravioletti durante la selezione compagno.

Metodi per la determinazione strutture UV-riflettenti e determinare la loro importanza nella Mate Preferenze utilizzando il Sailfin Molly<em&gt; Poecilia latipinna</em

Many organisms use cues and signals beyond human sensitivity during social interactions. It is important to take into account how organisms perceive their ...

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Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

6.5K Views.  Ohio Wesleyan University.  Questo protocollo descrive l'uso di spettrofotometria di rilevare strutture ultravioletti che riflette sugli organismi (in questo esempio, il sailfin Molly Poecilia latipinna) e descrive test a scelta dicotomiche per i pesci che permettono inferenze da apportare sul ruolo di spunti ultravioletti durante la selezione compagno. 

Video: Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female&#39;s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George&#39;s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Fishery-induced changes to exploited crustacean fisheries, such as the American lobster fishery, could potentially influence their reproductive dynamics, leading to a reduction in mating success. This study&#39;s goal was to develop a noninvasive method for ascertaining the mating success of female lobsters that may be physiologically or functionally mature.

Un metodo non invasivo per<em&gt; In situ</em&gt; Determinazione del successo di accoppiamento in americano femminile Aragoste (<em&gt; Homarus americanus</em&gt;)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

Ricerca

Ambiente

Multimodal Optical Microscopy Methods Reveal Polyp Tissue Morphology and Structure in Caribbean Reef Building Corals

An integrated suite of imaging techniques has been applied to determine the three-dimensional (3D) morphology and cellular structure of polyp tissues comprising the Caribbean reef building corals Montastraeaannularis and M. faveolata. These approaches include fluorescence microscopy (FM), serial block face imaging (SBFI), and two-photon confocal laser scanning microscopy (TPLSM). SBFI provides deep tissue imaging after physical sectioning; it details the tissue surface texture and 3D visualization to tissue depths of more than 2 mm. Complementary FM and TPLSM yield ultra-high resolution images of tissue cellular structure. Results have: (1) identified previously unreported lobate tissue morphologies on the outer wall of individual coral polyps&#160;and (2) created the first surface maps of the 3D distribution and tissue density of chromatophores and algae-like dinoflagellate zooxanthellae endosymbionts. Spectral absorption peaks of 500 nm and 675 nm, respectively, suggest that M. annularis and M. faveolata contain similar types of chlorophyll and chromatophores. However, M. annularis and M. faveolata exhibit significant differences in the tissue density and 3D distribution of these key cellular components. This study focusing on imaging methods indicates that SBFI is extremely useful for analysis of large mm-scale samples of decalcified coral tissues. Complimentary FM and TPLSM reveal subtle submillimeter&#160;scale changes in cellular distribution and density in nondecalcified coral tissue samples. The TPLSM technique affords: (1) minimally invasive sample preparation,&#160;(2) superior optical sectioning ability,&#160;and (3) minimal light absorption and scattering, while still permitting deep tissue imaging.

An integrated suite of imaging techniques has been applied to determine polyp morphology and tissue structure in the Caribbean corals Montastraeaannularis and M. faveolata. Fluorescence, serial block face,&#160;and two-photon confocal laser scanning microscopy have identified lobate structure, polyp walls, and estimated chromatophore and zooxanthellae densities and distributions.

Multimodali ottici Metodi Microscopia Reveal Polyp Tissue morfologia e struttura in Caribbean Reef costruzione Coralli

An integrated suite of imaging techniques has been applied to determine the three-dimensional (3D) morphology and cellular structure of polyp tissues ...

The Identification of Sea Lamprey Pheromones Using Bioassay-Guided Fractionation

Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and identification of an active pheromone compound. This method has resulted in the successful characterization of the chemical signals that function as pheromones in a wide range of animal species. Sea lampreys rely on olfaction to detect pheromones that mediate behavioral or physiological responses. We use this knowledge of fish biology to posit functions of putative pheromones and to guide the isolation and identification of active pheromone components. Chromatography is used to extract, concentrate, and separate compounds from the conditioned water. Electro-olfactogram (EOG) recordings are conducted to determine which fractions elicit olfactory responses. Two-choice maze behavioral assays are then used to determine if any of the odorous fractions are also behaviorally active and induce a preference. Spectrometric and spectroscopic methods provide the molecular weight and structural information to assist with the structure elucidation. The bioactivity of the pure compounds is confirmed with EOG and behavioral assays. The behavioral responses observed in the maze should ultimately be validated in a field setting to confirm their function in a natural stream setting. These bioassays play a dual role to 1) guide the fractionation process and 2) confirm and further define the bioactivity of isolated components. Here, we report the representative results of a sea lamprey pheromone identification that exemplify the utility of the bioassay-guided fractionation approach. The identification of sea lamprey pheromones is particularly important because a modulation of its pheromone communication system is among the options considered to control the invasive sea lamprey in the Laurentian Great Lakes. This method can be readily adapted to characterize the chemical communication in a broad array of taxa and shed light on waterborne chemical ecology.

Here, we present a protocol to isolate and characterize the structure, olfactory potency, and behavioral response of putative pheromone compounds of sea lampreys.

L'identificazione di feromoni Petromyzon marinus mediante frazionamento analisi biologica-Guida

Bioassay-guided fractionation is an iterative approach that uses the results of physiological and behavioral bioassays to guide the isolation and ...

Biochimica

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a potential mate to form a pair. While this phenomenon has attracted interest of researchers, methods for its analysis have not been well described. Here we describe procedures for: 1) individual culturing and staging of Tigriopus juveniles and adults, and 2) video-based analysis of their mate-guarding behavior. The culturing method enables experimental control of paring experience of animals as well as the ability to track their development before behavioral tests. The analysis method allows quantitative evaluation of several aspects of the mate-guarding behavior, including capturing attempts by males and swimming trajectory of mate-guarding pairs. Although these methods were originally established for ethological studies on Tigriopus, with proper modifications they can also be applied to studies of other zooplankton in different research fields, such as physiology, toxicology, and ecological genetics.

Individual Culturing of Tigriopus Copepods and Quantitative Analysis of Their Mate-guarding Behavior

Mate-guarding behavior plays an important role in reproduction of intertidal copepods of the genus Tigriopus. However, methods for studying this behavior have not been well described. Here we describe methods for: 1) individual culture of virgin Tigriopus animals, and 2) quantitative analysis of their mate-guarding behavior.

Coltura individuali di copepodi Tigriopus e analisi quantitativa del loro comportamento Mate-guardia

Copepods of the genus Tigriopus, which are common zooplankton in rocky tide pools, show precopulatory mate-guarding behavior where a male clasps a ...

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the appearance and behavior of visual stimuli, compared to manipulating live animals. Here, we present the FishSim Animation Toolchain, a software framework developed to provide researchers with an easy-to-use method for implementing 3D computer animations in behavioral experiments with fish. The toolchain offers templates to create virtual 3D stimuli of five different fish species. Stimuli are customizable in both appearance and size, based on photographs taken of live fish. Multiple stimuli can be animated by recording swimming paths in a virtual environment using a video game controller. To increase standardization of the simulated behavior, the prerecorded swimming path may be replayed with different stimuli. Multiple animations can later be organized into playlists and presented on monitors during experiments with live fish.
In a case study with sailfin mollies (Poecilia latipinna), we provide a protocol on how to conduct a mate-choice copying experiment with FishSim. We utilized this method to create and animate virtual males and virtual model females, and then presented these to live focal females in a binary choice experiment. Our results demonstrate that computer animation may be used to simulate virtual fish in a mate-choice copying experiment to investigate the role of female gravid spots as an indication of quality for a model female in mate-choice copying.
Applying this method is not limited to mate-choice copying experiments but can be used in various experimental designs. Still, its usability depends on the visual capabilities of the study species and first needs validation. Overall, computer animations offer a high degree of control and standardization in experiments and bear the potential to &#39;reduce&#39; and &#39;replace&#39; live stimulus animals as well as to &#39;refine&#39; experimental procedures.

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Using the novel FishSim Animation Toolchain, we present a protocol for non-invasive visual manipulation of public information in the context of mate-choice copying in sailfin mollies. FishSim Animation Toolchain provides an easy-to-use framework for the design, animation and presentation of computer-animated fish stimuli for behavioral experiments with live test fish.

Utilizzando la Toolchain di animazione FishSim per indagare il comportamento dei pesci: un caso di studio su Mate-scelta copia In Mollies Sailfin

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the ...

Comportamento

Behavioral Tracking and Neuromast Imaging of Mexican Cavefish

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among these traits, foraging behavior is one of the useful windows into functional advantages of behavioral trait evolution. Presented herein are updated methods for analyzing vibration attraction behavior (VAB: an adaptive foraging behavior) and imaging of associated mechanosensors of cave-adapted tetra, Astyanax mexicanus. In addition, methods are presented for high-throughput tracking of a series of additional cavefish behaviors including hyperactivity and sleep-loss. Cavefish also show asociality, repetitive behavior and higher anxiety. Therefore, cavefish serve as an animal model for evolved behaviors. These methods use free-software and custom-made scripts that can be applied to other types of behavior. These methods provide practical and cost-effective alternatives to commercially available tracking software.

Here, we present methods for high-throughput study of a series of the Mexican cavefish behaviors and vital staining of a mechanosensory system. These methods use free-software and custom-made scripts, providing a practical and cost-effective method for the studies of behaviors.

Rilevamento comportamentale e formazione immagine di Neuromast della caverna messicano

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among ...

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response is a vision-based, reflexive behavior that has been observed in all vertebrates tested. Tadpole eyes are small so the tail flip response was used as alternative measure, which requires a trained technician to record the subtle response. We developed an alternative behavior assay based on the fact that tadpoles prefer to swim on the white side of a tank when placed in a tank with both black and white sides. The assay presented here is an inexpensive, simple alternative that creates a response that is easily measured. The setup consists of a tripod, webcam and nested testing tanks, readily available in most Xenopus laboratories. This article includes a movie showing the behavior of tadpoles, before and after severing the optic nerve. In order to test the function of one eye, we also include representative results of a tadpole in which each eye underwent retinal axotomy on consecutive days. Future studies could develop an automated version of this assay for testing the vision of many tadpoles at once.

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis

Xenopus laevis tadpoles prefer swimming on the white side of a black/white tank. This behavior is guided by their vision. Based on this behavior, we present a simple assay to test the visual function of tadpoles.

Un test comportamentale semplice for Testing Funzione visiva in<em&gt; Xenopus laevis</em

Measurement of the visual function in the tadpoles of the frog, Xenopus laevis, allows screening for blindness in live animals. The optokinetic response ...

Neuroscienze

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

L&#39;audacia, l&#39;aggressività e saggi shoaling per Zebrafish comportamentali Sindromi

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Medicina

Analysis of Skeletal Muscle Defects in Larval Zebrafish by Birefringence and Touch-evoke Escape Response Assays

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular dystrophies1-3, congenital myopathies4,5, and disruptions in sarcomeric assembly6,7, due to high genomic and structural conservation with mammals8. Muscular disorganization and locomotive impairment can be quickly assessed in the zebrafish over the first few days post-fertilization. Two assays to help characterize skeletal muscle defects in zebrafish are birefringence (structural) and touch-evoked escape response (behavioral).
Birefringence is a physical property in which light is rotated as it passes through ordered matter, such as the pseudo-crystalline array of muscle sarcomeres9. It is a simple, noninvasive approach to assess muscle integrity in translucent zebrafish larvae early in development. Wild-type zebrafish with highly organized skeletal muscle appear very bright amidst a dark background when visualized between two polarized light filters, whereas muscle mutants have birefringence patterns specific to the primary muscular disorder they model. Zebrafish modeling muscular dystrophies, diseases characterized by myofiber degeneration followed by repeated rounds of regeneration, exhibit degenerative dark patches in skeletal muscle under polarized light. Nondystrophic myopathies are not associated with necrosis or regenerative changes, but result in disorganized myofibers and skeletal muscle weakness. Myopathic zebrafish typically show an overall reduction in birefringence, reflecting the disorganization of sarcomeres.
The touch-evoked escape assay involves observing an embryo&#39;s swimming behavior in response to tactile stimulation10-12. In comparison to wild-type larvae, mutant larvae frequently display a weak escape contraction, followed by slow swimming or other type of impaired motion that fails to propel the larvae more than a short distance12. The advantage of these assays is that disease progression in the same fish type can be monitored in vivo for several days, and that large numbers of fish can be analyzed in a short time relative to higher vertebrates.

The zebrafish is now an established and powerful tool for modeling muscular dystrophies, congenital myopathies, and related neuromuscular diseases. Birefringence and touch-evoked escape behavior are two common noninvasive assays used to determine the degree of muscular disorganization and locomotive impairment of zebrafish embryos during early development.

Analisi dei difetti muscolo scheletrico larvale Zebrafish da Birifrangenza e Touch-evocare Fuga risposta Saggi

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular ...

Biologia

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump&#39;s bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

This protocol describes the use of sump swabs and sludge analysis of zebrafish systems, which leads to increased detection compared to the sole use of sentinels to detect pathogens such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa. A system to monitor P. tomentosa eggs in quarantine is also proposed.

Metodi per la determinazione strutture UV-riflettenti e determinare la loro importanza nella Mate Preferenze utilizzando il Sailfin Molly

Riepilogo

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Capitoli in questo video

Un metodo non invasivo per

Multimodali ottici Metodi Microscopia Reveal Polyp Tissue morfologia e struttura in Caribbean Reef costruzione Coralli

L'identificazione di feromoni Petromyzon marinus mediante frazionamento analisi biologica-Guida

Coltura individuali di copepodi Tigriopus e analisi quantitativa del loro comportamento Mate-guardia

Utilizzando la Toolchain di animazione FishSim per indagare il comportamento dei pesci: un caso di studio su Mate-scelta copia In Mollies Sailfin

Rilevamento comportamentale e formazione immagine di Neuromast della caverna messicano

Un test comportamentale semplice for Testing Funzione visiva in

L'audacia, l'aggressività e saggi shoaling per Zebrafish comportamentali Sindromi

Analisi dei difetti muscolo scheletrico larvale Zebrafish da Birifrangenza e Touch-evocare Fuga risposta Saggi

Selezione ambientale di hydrophila dell'aeromonas, Mycobacterium spp. e Pseudocapillaria tomentosa in Zebrafish sistemi