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.

Todos os experimentos foram realizados com a aprovação do Comitê de Cuidado e Uso Institucional Animal da Ohio Wesleyan University. 1. Gravação UV refletância dos peixes usando um espectrofotômetro <ol><li> Calibrar um espectrofotômetro e fonte de luz com um padrão branco conhecido em toda a gama de comprimentos de onda a ser medido de acordo com as recomendações do instrumento ou software. </li><li> Anestesiar os peixes, colocando em uma solução a 0,5% de acetato de 3 aminobenzoate-metano sal de ácido sulfônico (MS-222) tamponada com igual quantidade de bicarbonato de sódio até que o peixe deixa de responder. Uma vez que não responde, imediatamente colocar o peixe em um fundo preto, não-reflexivo. Nota: MS-222 é potencialmente tóxico, e devem ser usadas luvas em todos os momentos. </li><li> Fazer todas as medições em uma sala escura para minimizar o excesso de luz além do que a fonte de luz espectrômetro. </li><li> reflectância medida usando uma sonda de fibra óptica mantida contra o corpo do organismo. FaçoCertifique-se para formar um ângulo de 45 ° consistente entre as fibras ópticas e do corpo, como é construído em alguns modelos de sonda. Nota: Para manter a consistência de medição, pode ser desejável a média em vários scans (2 - 5) por região do corpo. </li><li> Gravar dados de reflectância com software de análise de reflectância apropriada. Nota: O software utilizado aqui foi definido como: integração em tempo = 2,000 seg, média scans = 2, e de pixel suavização = 5. Os dados foram copiados e colados em uma planilha. </li><li> Repetir em várias áreas do corpo para determinar a presença ou ausência de reflectância de UV e gama de variação fenotípica entre organismos. Nota: exames preliminares podem ser necessários para determinar qual, se houver, áreas do corpo mostram consistentemente marcações UV. </li><li> Devolver o peixe para um aquário de exploração bem oxigenado até que esteja totalmente recuperado, como evidenciado por batidas opérculo normais e comportamentos de natação. Nota: um protector comercial para restaurar o revestimento mucoso para o corpo podem ser adicionados para ajudar na recuperação da pele. </li><li> Repcomer em vários indivíduos, incluindo ambos os sexos, se desejar. </li><li> Na planilha, onde todos os dados foram copiados, os dados do gráfico usando um gráfico de dispersão, com reflectância no eixo Y e comprimento de onda no eixo-X. picos de amplitude na gama dos UV (300-400 nm) indicar a presença de reflectância de UV sobre o peixe. </li></ol> 2. Os ensaios de acasalamento dicotômicas Nota: Execute observações de controle, em que os peixes focal, filtros UV e filtros opacos estão no local. Nestes ensaios, quando os filtros são removidos opacas, peixes focal será exposto a partes do tanque indicam a presença e ausência de luz UV, mas que não possuem parceiros de acasalamento potenciais. Para mais detalhes sobre a validade dos testes de escolha dicotômicas neste experimento ver Palmer e Hankison (2015) 10, embora note que estes testes também têm desvantagens 11. <ol><li> Use indivíduos sexualmente receptivas como organismos focais. Use feminina P. latipinna &lt;48 horas pós-parto 5 aND machos isolado para, pelo menos, 24 horas antes dos testes para melhorar a probabilidade de motivação sexual. </li><li> Coloque um único indivíduo focal em uma área neutra adjacente a dois compartimentos de escolha. Para P. latipinna, usar um aquário teste 75,7 L (30,5 cm x 30,5 cm x 75 cm; Figura 1), dividido em três secções por meio de divisórias de Plexiglas transparente, formando uma área central para o indivíduo focal flanqueado por dois compartimentos de igual tamanho de escolha. Certifique-se de que nenhuma água ou olfativas pistas são compartilhadas entre as seções de tanques individuais. Repita o procedimento para cada indivíduo a ser observado. </li></ol><img alt="figura 1" src="/files/ftp_upload/54453/54453fig1.jpg" /> Figura 1. Experimental Aquarium Setup Um aquário retangular dividido em três seções:. Uma área central que realizou o indivíduo focal e dois compartimentos finais que realizou pares de objetos (a figura mostra uma mulher no meio com machos de objeto nas extremidades). fil intercambiáveisters que ou reflectância UV bloqueado ou permitido poderia ser colocada sobre o divisórias de vidro que dividem as seções masculina e feminina do aquário. Esta figura é modificada a partir Palmer e Hankison (2015) 10. <a href="http://ecsource.jove.com/files/ftp_upload/54453/54453fig1large.jpg" target="_blank">Por favor clique aqui para ver uma versão maior desta figura.</a> <ol start="3"><li> Sob condições de iluminação de espectro total (iluminação cujo espectro se estende para o UV-A gama), isolar um parceiro de acasalamento potencial em cada compartimento final (compartimento escolha) do aquário teste. Garantir que os potenciais parceiros de acasalamento se parecem entre si, tanto quanto possível, em tamanho, coloração e exibir características, de modo que a principal diferença entre os dois parceiros de acasalamento isoladas é se eles aparecem atrás de um filtro que bloqueia ou transmite a luz UV. NOTA: Pode ser preferível para gravar reflectância no espectro visível para permitir a melhor correspondência de cores possível, alembora aqui correspondente foi feito a olho nu. </li><li> Aclimatar indivíduos sob isolamento visuais (filtros opacos) por 15 min. Assegure-se que + e UV - filtros estão no lugar neste momento. Na sequência de aclimatação, remova os filtros opacos. Para assegurar que os desvios laterais não estão presentes (preferências inerente de um dos lados do tanque, independentemente do objecto), atribua aleatoriamente a + UV e UV - lados do tanque para cada cada ensaio. </li><li> Grave a quantidade de tempo que o indivíduo focal passa dentro de zonas preferenciais específicas (áreas próximas aos compartimentos de escolha, neste caso cerca de dois corpos padrão comprimentos) usando cronômetros. No caso do peixe fêmea, isto é a quantidade de tempo que passa perto de cada um dos dois machos, e vice-versa para peixes focal do sexo masculino. NOTA: O software gravador de eventos também pode ser utilizado para gravação. Quinze minutos ensaios foram utilizados neste experimento e os peixes que não visitar cada zona de preferência pelo menos uma vez foram excluídos dos resultados como sendo sem resposta. </li><li> Registar e analisar o tempo que os peixes passam quase vazio UV + e UV - compartimentos para garantir que não há um viés inerente para um ambiente de iluminação especial que podem influenciar a escolha de acasalamento dos peixes. Use testes t pareados para determinar se os indivíduos preferem uma condição UV particular. </li></ol>

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The authors have nothing to disclose.

Espectrofotometria foi bem sucedida na identificação de marcas UV em P. latipinna. Ambos os sexos de P. latipinna possuem marcações UV ao longo de seus lados. Além disso, alguns machos tinha marcas UV em suas barbatanas dorsais, traços encontrados anteriormente a ser importante nas preferências de acasalamento femininos 7. Recomendamos a utilização de espectrofotometria de UV como um mecanismo para detectar a presença de marcas UV. Mais testes poderia d...

Compreender os sinais e sinais utilizados nas interações sociais animais nos permite compreender a variação fenotípica dentro e entre as espécies. Esta variação desempenha um papel importante nos processos evolutivos, tais como a divergência populacional, a selecção sexual, e especiação. Muitas vezes, porém, os pesquisadores estão limitados a explorar as pistas mais óbvias para sistemas sensoriais humanos, especialmente aqueles dentro dos domínios visuais ou auditivas. Uso de espectrofotometria, no entanto, nos permite expandir nossas investigações para além do espectro visível humana e em comprimentos de onda que podem ser importantes em interações sociais em outras espécies. Em particular, a comunicação de curto alcance proporcionada por luz ultravioleta (UV; 300-400 nm) sensibilidade tem o potencial de ser altamente vantajoso durante a escolha do parceiro 1. Muitos predadores visualmente caça de aves e peixes, por exemplo, não são capazes de detectar a radiação UV. Em sistemas onde os machos mostrar elaboradamentepara fêmeas, esses machos reduziria o risco de predação, mantendo a sua capacidade de atrair parceiros, explorando o espectro UV em vez de desenvolver sinais detectáveis ​​no espectro visível 2,3 .. Se alguém deixa de considerar a possibilidade de que os organismos estão se comunicando uns com os outros usam esses &quot;canais de comunicação privados&quot;, motoristas significativas de comportamento e evolução pode ser desperdiçada. Este protocolo descreve uma investigação sobre o uso de pistas de UV para a escolha do companheiro na sailfin molly, Poecilia latipinna, um peixe polígamos que não conheceu anteriormente capacidade de detectar UV ou utilizar marcações UV. Esta espécie de peixe tem uma proximidade filogenética perto de outros peixes Vivípara sensíveis aos raios UV 4 e há evidências de que microspectroscopic P. latipinna, juntamente com outras espécies tais como P. Molly mexicana e P. formosa, possuem uma classe de cones (células fotorreceptoras responsáveis ​​pela covisão lor), que são mais sensíveis para comprimentos de onda UV 5. Nesta espécie dimorfismo sexual, escolha feminina tem desempenhado um papel importante na evolução da dos machos barbatanas coloridas e ampliadas brilhantemente 6-9. Esta metodologia permite-nos explorar se UV é um meio adicional pelo qual as fêmeas avaliar a qualidade do sexo masculino. A detecção e medição de marcações UV sobre P. latipinna usando um espectrofotómetro com uma sonda de fibra óptica é detalhado aqui. Além disso, se mollies fêmeas receptivas diferencialmente associar com machos vistos através de um filtro óptico de transmissão de luz de espectro total, incluindo UV-A ([UV +]; 320-700 nm) e os machos vistos através de um filtro anti-UV ([UV]; 400 - 700 nm) é discutida. Este método tem amplas aplicações para descobrir padrões de sensibilidade UV e cor em peixes e outros organismos, permitindo a investigação sobre uma variedade de questões que envolvem UV e seu papel no 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

Muitos organismos usam pistas e sinais para além sensibilidade humana durante as interações sociais. É importante levar em conta a forma como os organismos percebem seus mundos ao tentar compreender o seu comportamento e ecologia. Sensibilidade ao espectro de ultravioleta (UV; 300-400 nm) é encontrado através de vários géneros de aves, peixes, répteis, anfíbios e até mamíferos. Este protocolo descreve uma técnica pela análise de organismos para a presença de estruturas de reflexão de UV e um método para testar se estes sinais são utilizados como sinais de sociais no contexto da escolha de parceiros. Um espectrofotómetro é utilizado para detectar a presença de reflectância de UV e a variação na intensidade reflexiva entre os indivíduos e os sexos. Um exemplo desta técnica é apresentada em que um teste de escolha do parceiro dicotómica expõe indivíduos sexualmente receptiva a indivíduos do sexo oposto cuja aparência visual pode ser manipulado por meio de filtros que transmitem, quer do espectro de comprimentos de onda de UV ou bloco completo. estesistema permitiu a determinação que as mulheres, mas não do sexo masculino, mollies sailfin (Poecilia latipinna) estavam usando marcas de UV como parte de suas decisões de acasalamento. Estes tipos de estudos servem para ampliar o conhecimento da gama de organismos que utilizam UV e fornecer insights sobre como UV desempenha um papel em suas vidas.

<html> As Figuras 1 e 2 mostram a preferência do aquário de acasalamento configurar e locais de medição UV para nossos experimentos. Medição da reflectância de UV permitiu a determinação de que a P. latipinna possuem características de UV, em especial ao longo dos lados do seu corpo (Figura 3), para além da variação individual em esses traços. Um...

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

Este protocolo descreve o uso de espectrofotometria para detectar estruturas de reflexão de raios ultravioleta sobre os organismos (neste exemplo, o sailfin molly Poecilia latipinna) e descreve testes de escolha dicotômicas para peixes que permitem inferências a serem feitas sobre o papel das pistas ultravioleta durante a escolha do parceiro.

Ensaios para detectar estruturas de reflexão de UV e determinar a sua importância na preferência do companheiro usando o 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 visualizações.  Ohio Wesleyan University.  Este protocolo descreve o uso de espectrofotometria para detectar estruturas de reflexão de raios ultravioleta sobre os organismos (neste exemplo, o sailfin molly Poecilia latipinna) e descreve testes de escolha dicotômicas para peixes que permitem inferências a serem feitas sobre o papel das pistas ultravioleta durante a escolha do parceiro. 

Vídeo: 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.

Um método não invasivo para<em&gt; Em situ</em&gt; Determinação do sucesso de acasalamento em fêmeas lagostas americanas (<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 ...

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.

Multimodal ópticos métodos de microscopia Revelar Polyp Tissue Morfologia e Estrutura em Caribe Reef construção Corais

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.

A identificação das feromonas de lampreia do mar usando guiada por bioensaio fracionamento

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

Bioquímica

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.

Cultivo individual de copépodes Tigriopus e análise quantitativa de seu comportamento companheiro-guarda

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.

Usando o conjunto de ferramentas de animação de FishSim para investigar o comportamento dos peixes: um estudo de caso na escolha do companheiro copiando em 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.

Acompanhamento comportamental e Neuromast imagem de Cavefish mexicano

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.

Um Ensaio Comportamental simples para Teste de Função Visual em<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 ...

Neurociência

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.

Arrojo, agressão e Ensaios shoaling para Zebrafish Síndromes Comportamentais

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.

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.

Ensaios para detectar estruturas de reflexão de UV e determinar a sua importância na preferência do companheiro usando o Sailfin Molly

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Um método não invasivo para