The authors have no acknowledgements.

Dichiarazione etica: Tutte le procedure sono state approvate dal comitato della University of Sydney Etica Animale ed adeguato alle Sanitario Nazionale e medicina Research Council di linee guida in Australia. Le procedure sono state effettuate su topi C57 / BL6J che sono stati allevati presso l&#39;Università di Sydney Bosch roditore strumento. Tutti i topi sono stati alloggiati in un unico locale sufficientemente ventilato in 21 ° C di temperatura ambiente in un ciclo luce-buio 12 ore con le luci alle 0600 ore in gabbie ventilazione individuale con ad libitum l&#39;accesso al cibo secco e acqua. Femmine tardo-gravidanza sono stati assegnati in modo casuale a condizioni di alloggio standard o arricchito l&#39;ambiente. 1. Housing (livelli di arricchimento): <ol><li> Ottenere 4 tarda gravidanza adulti topi femmina. Casualmente assegna 2 per la condizione standard e porre ciascuno di essi in una gabbia mouse standard pulito (ingombro 391 x 199 x 160 mm), contenente un mouse igloo rosso. Per la condizione arricchito, posizionare il remavorazio 2 topi in una singola gabbia topo-sized clean (ingombro 462 x 403 x 404 mm). </li><li> Nella gabbia di arricchimento, posizionare una varietà di oggetti progettati per aumentare la stimolazione sensoriale e motoria (ad esempio ruote corsa, stimoli visivi, batuffoli di cotone profumate, velcro). </li><li> Ogni 2 o 3 giorni si muovono questi oggetti circa all&#39;interno della gabbia; aggiornare qualsiasi che sono stati distrutti. </li><li> Dopo lo svezzamento a 21 giorni dopo la nascita, gli animali di sesso e il luogo in alloggi separati maschio-femmina in linea con le condizioni ambientali che sono state sollevate. Per la condizione arricchito in modo che ci sono tra i 3 ei 10 topi per gabbia con 2 e 5 topi per lo standard. Iniziare i test comportamentali una volta raggiunta l&#39;età adulta animali (12-14 settimane di età). </li></ol> 2. La costruzione del puzzle Box <ol><li> Ottenere 6 parti di acrilico bianco (o altro materiale non poroso): uno 750 x 280 mm, due 280 x 250 mm, due 750 x 250 mm, ed uno 150 x 280 mm (vedi figura 1). </li><li> Ottenere un pezzo di acrilico nero 280 x 250 mm, con una apertura quadrata di 40 x 40 mm tagliato in un lato di questo pezzo (vedi Figura 1). </li><li> Assemblare il puzzle Box come segue: usare il pezzo mm 750 x 280 come il fondo della scatola, utilizzare 280 x 250 mm pezzi come le estremità della scatola, ed i pezzi da 750 x 250 mm come lati della scatola. </li><li> Misurare 150 millimetri nella casella da un&#39;estremità, e posizionare il pezzo nero di acrilico attraverso la scatola in modo che si divide in due scomparti (uno grande, uno piccolo) con il filo apertura con il fondo della scatola. </li><li> Prendete il pezzo di mm acrilico bianco 150 x 280 e posizionarlo in cima al vano più piccolo della scatola assicurandosi che copre l&#39;area completamente, fornendo una camera oscura &quot;goal-box&quot;. Applicare questo pezzo di acrilico al corpo scatola principale cerniere, o lasciare libero di essere completamente rimosso durante la prova comportamentale. </li><li> Prendere 3 pezzi di acrilico (tre 4 x 120 mm) e unisciti a fare una &quot;u-shapchannel ed &quot;. </li></ol><img alt="Figura 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225fig1highres.jpg" width="500" /> Figura 1: Schema del compito Puzzle Box. Il Puzzle Box è una scatola acrilica costituito da uno spazio aperto-campo (600 x 280 mm) e una zona riparata goal-box (150 x 280 mm), che misura 750 x 280 mm di tutti. Una apertura 40 x 40 mm all&#39;interno della partizione (grigio) che separa le due aree permette animali di accedere alla zona obiettivo-scatola coperta dal campo aperto. Questa apertura è bloccata con ostacoli sempre più difficili da rimuovere come test progredisce. Gli animali sono sottoposti ad un protocollo di cinque giorni, che consiste di quattro condizioni ostruzione con tre prove per ogni condizione. 3. Esecuzione della Task Puzzle Box <ol><li> Pulire accuratamente la scatola di puzzle con il 70% di alcol. Ripetere questo passaggio tra ogni testati su animali. </li><li> Posizionare un pulito il mouse igloo rosso nel ar obiettivo-boxEA del Puzzle Box, e sostituire il coperchio sul gol-box. </li><li> Se vi è una condizione di ostruzione in fase di test, posizionare l&#39;ostruzione all&#39;interno della soglia della meta-box (vedi figura 2). </li><li> Posizionare il mouse in fase di test nella sezione in campo aperto del Puzzle Box, orientato verso la meta-box, e alla più lontana dalla meta-box. </li><li> Registrare il tempo impiegato per tutte e quattro le zampe dell&#39;animale per entrare nella sezione obiettivo-box del Puzzle Box. </li><li> Se un animale non entra in porta-box, interrompere il processo una volta raggiunto il limite di tempo (vedi tabella 1). </li><li> Una volta che una prova è terminato, rimuovere l&#39;animale dalla Puzzle Box e metterlo in una gabbia un&#39;azienda distinta fino all&#39;inizio della prova successiva. Mantenere un gap 60 a 180 sec tra processi per ciascun animale. </li><li> Per ciascun animale, effettuare tre prove al giorno per cinque giorni consecutivi di test, con quattro condizioni ostruzione e tre prove di ciascuna condizione. Il terzo triAl di una data condizione ostruzione deve sempre somministrato su un giorno successivo (vedi Tabella 1). </li></ol><img alt="Tabella 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225table1.jpg" width="500" /> Tabella 1: Schema del compito Puzzle Box. Il compito Puzzle Box viene eseguita per cinque giorni, e consiste di quattro condizioni ostruzione. Ci sono tre prove su ciascuno dei primi quattro giorni, e uno il quinto giorno. Ogni condizione di ostruzione ha tre prove; le prime due in un giorno, e il terzo giorno immediatamente successivo. La prima prova di una condizione di ostruzione mira a testare native capacità di risolvere problemi, il secondo processo esamina acquisizione compito e rinforzo, e la terza prova è usato come un test per la conservazione della soluzione e di richiamo. <img alt="Figura 2" fo:content-width="6in" src="/files/ftp_upload/52225/52225fig2highres.jpg" width="600" /> Figura 2: ostruirecondizioni di ioni all&#39;interno del compito Puzzle Box. Diagrammi di dell&#39;arena Puzzle Box e le condizioni di ostruzione utilizzati in questo studio. (A) Condizione 0 (C0), senza ostruzioni presenti all&#39;interno della porta tra il campo aperto e aree goal-box. (B) Condizione 1 (C1 ), con un canale presente &quot;U&quot; all&#39;interno della porta tra il campo aperto e aree obiettivo-box. (C) Condizione 2 (C2), in cui il canale è riempito con lettiera pulita. (D) Condizione 3 (C3 ), con un &quot;plug tissue&quot; presente nella porta tra il campo aperto e aree obiettivo-box. (E) Condizione 4 (C4) con un &quot;plug foam&quot; presente nella porta tra il campo aperto e meta-box aree. Dimensioni dell&#39;arena sono secondo elencato in Metodi e Figura 1. 4. Interpretazione dei dati <ol><li> Rappresentare i dati sia come tempo necessario per completare il processo (comprese le prove nulli in cui gli animali non è stata completata l&#39;operazione entro il termine previsto) o come il numero di prove nulli. </li><li> Utilizzare una misure ripetute ANOVA per valutare l&#39;effetto della condizione abitativa, all&#39;andamento nel Puzzle Box, con il tipo di ostruzione / condizioni (C) e il numero di task (T) entro soggetti fattori e il livello di arricchimento (standard contro arricchito) tra i soggetti-factor . </li></ol>

The authors declare they have no competing financial interests.

I dati presentati dimostrano che il Puzzle Box può essere utilizzato in modo efficace per valutare l&#39;impatto di EE. Topi allevati in ambienti arricchiti sempre preso molto meno tempo per risolvere i puzzle di ostruzione all&#39;interno di questa analisi comportamentale che ha animali allevati all&#39;interno di condizioni standard di laboratorio. Inoltre, questa differenza era più importante nella prima prova per tre delle quattro condizioni testate, suggerendo EE ha una maggiore influenza sulla native capacità d...

L&#39;arricchimento ambientale (EE) può essere definita come un ambiente che forniscono gli animali con maggiori opportunità di interazione sociale, attività motoria, e una maggiore stimolazione sensoriale del solito vissuto in un environment.EE standard di laboratorio ha dimostrato di influire in modo coerente il comportamento degli animali, determinando modifiche come la riduzione dello stress e l&#39;ansia di attività legate 8-10, il miglioramento delle prestazioni in compiti di apprendimento e memoria 8,11, insorgenza precoce di coordinazione motoria e l&#39;attività esplorativa 11, cambiamenti di cure materne 8 nonché resistenza alle sostanze stupefacenti 12- 15. Inoltre, EE è stato rivelato per migliorare gli effetti delle malattie neurodegenerative, ritardando l&#39;insorgenza e diminuire la gravità dei sintomi in modelli animali di Huntington di 1-4,16, Parkinson 17 e malattia di Alzheimer 18. Questi cambiamenti COrrelazione con le alterazioni anatomiche e molecolari EE è noto per indurre in tutto il cervello. Animali allevati in ambienti arricchiti dalle prime fasi di sviluppo mostrano una miriade di cambiamenti neurali, tra cui un aumento di peso del cervello e lo spessore corticale, dendritica ramificazione 9,2-22 e la densità sinaptica 23. EE può alterare sia il livello e la tempistica di espressione del fattore di crescita 9,24-30, che ha dimostrato di contribuire allo sviluppo accelerato sensoriale 25,26,28,29, mnemonico 30, nonché circuiti motori 31,32. Il lavoro precedente ha rivelato a risultati contraddittori momenti in cui si esamini l&#39;impatto di EE, senza tener conto dei diversi tipi di animali e ambienti utilizzati all&#39;interno dei singoli studi 9,24,27,30. Attualmente, non esiste compito comportamentale coerente e semplice che può essere utilizzato per misurare l&#39;efficacia di diversi paradigmi EE in ceppi diversi e specIES di animali. Il compito Puzzle Box è stato concepito come un semplice test per determinare problema nativa di un animale solving 7. Gli animali posti in zona aperta sono tenuti a rimuovere ostacolare materiali situati all&#39;interno di una piccola apertura per poter accedere a una coperta regione / rifugio. Ciascun soggetto è dato tre prove con lo stesso ostruzione per valutare tre diversi attributi cognitive. La prima prova si ottiene un indicazione di base di capacità di problem solving intrinseca o nativa. La seconda prova, eseguire lo stesso giorno, dà qualche indicazione della capacità dell&#39;animale di migliorare e rafforzare le strategie per rimuovere l&#39;ostruzione specifica in tal modo. La terza prova, condotta il giorno seguente, permette di comprendere meglio la capacità del soggetto di trattenere e richiamare la soluzione imparato al compito. La motivazione per risolvere questi &quot;enigmi ostruzione&quot; di animali può essere variata, potenzialmenteevocando un innato desiderio di evitare campi aperti e cercare riparo, così come un&#39;unità intrinseca di esplorare l&#39;ambiente circostante 6,7. La moltitudine di potenziali conducenti comportamentali alla base del desiderio di risolvere il puzzle Box suggerisce che diverse aree del cervello sono coinvolte nella mediazione prestazioni compito. Il lavoro precedente ha mostrato che in modelli murini di schizofrenia, corteccia prefrontale e l&#39;ippocampo sono coinvolti nell&#39;acquisizione di questo compito 5. Uno studio della lesione nei ratti ha rivelato anche un gran numero di regioni cerebrali coinvolte nella prestazione Puzzle Box, tra cui vari nuclei del talamo, l&#39;ipotalamo, il cervelletto, e le strutture limbiche. Insieme, questi risultati indicano che impegnarsi in questo problem solving compito comporta una serie di strutture neurali associate alla funzione cognitiva. Il Puzzle Box è stato utilizzato con successo per valutare la capacità di problem solving di topi, così come deficit cognitivi esposto da mmodelli di urina di schizofrenia 5-7. Prestazioni del compito ha dimostrato di essere altamente coerente, e correlare bene con risultati di altri test cognitivi comportamentali 6. L&#39;obiettivo di questo lavoro è stato quindi di adattare il compito Puzzle Box per diventare un mezzo semplice ed affidabile per determinare l&#39;efficacia di EE.

<table><tbody><tr><td>Black Acrylic Board</td><td></td><td></td><td>280 x 250 mm with a 40 x 40 mm &quot;door&quot; cut into one side</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>280 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>280 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>750 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>750 x 250 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td>150 x 280 mm</td></tr><tr><td>White Acrylic Board</td><td></td><td></td><td></td></tr><tr><td>Underpass</td><td></td><td></td><td>3 pieces of 40 x 120 mm plexiglass or acrylic</td></tr><tr><td colspan="4">Note: If unable to access acrylic board, plexiglass or similar non-porous material will suffice.</td></tr><tr><td>Webcam</td><td>Logitech</td><td>C210</td><td>Fix to roof with electrical tape. Alternatively, use a tripod.</td></tr><tr><td>VirtualDub v1.10.4</td><td>VirtualDub</td><td>N/A&amp;nbsp;</td><td>Software for recording behaviour videos. Input from webcam.</td></tr><tr><td>TopScan v 3.0</td><td>CleverSys Inc.</td><td>N/A&amp;nbsp;</td><td>Software for automated top-view tracking and analysis of mouse behaviour. Captured videos are analysed post-hoc.</td></tr></tbody></table>

the use of the puzzle box as a means of assessing the efficacy of environmental enrichment

Environmental enrichment can dramatically influence the development and function of neural circuits. Further, enrichment has been shown to successfully delay the onset of symptoms in models of Huntington&#8217;s disease 1-4, suggesting environmental factors can evoke a neuroprotective effect against the progressive, cellular level damage observed in neurodegenerative disorders. The ways in which an animal can be environmentally enriched, however, can vary considerably. Further, there is no straightforward manner in which the effects of environmental enrichment can be assessed: most methods require either fairly complicated behavioral paradigms and/or postmortem anatomical/physiological analyses. This protocol describes the use of a simple and inexpensive behavioral assay, the Puzzle Box 5-7 as a robust means of determining the efficacy of increased social, sensory and motor stimulation on mice compared to cohorts raised in standard laboratory conditions. This simple problem solving task takes advantage of a rodent&#8217;s innate desire to avoid open enclosures by seeking shelter. Cognitive ability is assessed by adding increasingly complex impediments to the shelter&#8217;s entrance. The time a given subject takes to successfully remove the obstructions and enter the shelter serves as the primary metric for task performance. This method could provide a reliable means of rapidly assessing the efficacy of different enrichment protocols on cognitive function, thus paving the way for systematically determining the role specific environmental factors play in delaying the onset of neurodevelopmental and neurodegenerative disease.

I risultati qui descritti sono un campione rappresentativo, con i dati provenienti dalle diverse coorti composte da diverse cucciolate. Tutti i test comportamentali è stato condotto tra il 0700 e 1100 ore, con ordine sperimentazione casuale di animali all&#39;interno di una coorte. Animali sollevate in un ambiente arricchito (n = 14, 7 femmine e 7 maschi) hanno significativamente meno tempo per risolvere i compiti ostruzione all&#39;interno della scatola Puzzle di quelle sollevate in un ambiente standard (n = 15, 7 fem...

Watch this Scientific Journal Video about The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment at JoVE.com

The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

Environmental enrichment provides a potential protective effect against neurodegenerative disorders. Currently, however, there is no easy way of determining the efficacy of enrichment procedures. This protocol describes a simple &#8220;Puzzle Box&#8221; method for assessing an animal&#8217;s cognitive function, in order to reveal the effectiveness of environmental enrichment.

L&#39;uso del Puzzle Box come un mezzo per valutare l&#39;efficacia di arricchimento ambientale

Environmental enrichment can dramatically influence the development and function of neural circuits.  Further, enrichment has been shown to successfully ...

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JoVE Journal

Behavior

11.7K Views.  University of Sydney. Environmental enrichment provides a potential protective effect against neurodegenerative disorders. Currently, however, there is no easy way of determining the efficacy of enrichment procedures. This protocol describes a simple “Puzzle Box” method for assessing an animal’s cognitive function, in order to reveal the effectiveness of environmental enrichment.

Video: The Use of the Puzzle Box as a Means of Assessing the Efficacy of Environmental Enrichment

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

The Hole Consiglio Modified - Behavior, Cognition e l&#39;interazione sociale in topi e ratti di misura

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

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Comportamento

A Protocol for Housing Mice in an Enriched Environment

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent studies demonstrate that environmental enrichment decreases adiposity, increases energy expenditure, resists diet induced obesity, and causes cancer remission and inhibition in mice. EE typically consists of larger living space, a variety of &#8216;toys&#8217; to interact with, running wheels, and can include a number of other novel environmental changes. All of this fosters a more complex social engagement, cognitive and physical stimulations. Importantly, the toy location and type of toy is changed regularly, which encourages the mice to adapt to a frequently changing and novel environment. Many variables can be manipulated in EE to promote health effects in mice. Thus these approaches are difficult to control and must be properly managed to successfully replicate the associated phenotypes. Therefore, the goal of this video is to demonstrate how EE is properly set up and maintained to assure a complex, challenging, and controlled environment so that other researchers can easily reproduce the protective effects of EE against obesity and cancer.

Environmental enrichment for mice requires a complex and challenging setup, as well as comprehensive husbandry and handling techniques to assure robust metabolic and anti-cancer effects in the mice. This protocol provides detailed procedures to reproduce the above mentioned effects in mice.

Un protocollo per Mice Housing in un ambiente arricchito

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent ...

An Operant Intra-/Extra-dimensional Set-shift Task for Mice

Alterations in executive control and cognitive flexibility, such as attentional set-shifting abilities, are core features of several neuropsychiatric diseases. The most widely used neuropsychological tests for the evaluation of attentional set-shifting in human subjects are the Wisconsin Card Sorting Test (WCST) and the CANTAB Intra-/Extra-dimensional set shift task (ID/ED). These tasks have proven clinical relevance and have been modified and successfully adapted for research in animal models. However, currently available tasks for rodents present several limitations, mainly due to their manual-based testing procedures, which are hampering translational advances in psychiatric medicine. To overcome these limitations and to better mimic the original version in primates, we present the development of a novel operant-based two-chamber ID/ED "Operon" task for rodents. We demonstrated the effectiveness of this novel task to measure different facets of cognitive flexibility in mice including attentional set formation and shifting, and reversal learning. Moreover, we show the high flexibility of this task in which three different perceptual dimensions can be manipulated with a high number of stimuli cues for each dimension. This novel ID/ED Operon task can be an effective preclinical tool for drug testing and/or large genetic screening relevant to the study of executive dysfunction and cognitive symptoms found in psychiatric disorders.

Attentional set-shifting cognitive abnormalities are a core feature of many psychiatric diseases. Here, we present the development of a novel automatic system for the effective study of attentional set-shifting abilities, executive functions and other cognitive abilities in mice with high translational validity to human studies.

Un operante intra / extra-dimensionale Set-shift attività per Mice

Alterations in executive control and cognitive flexibility, such as attentional set-shifting abilities, are core features of several neuropsychiatric ...

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

Esplorazione oggetto romanzo come un test Potenziale di ordine superiore ripetitivi comportamenti nei topi

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially.

The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice.

L&#39;introduzione di Clicker Training come un arricchimento conoscitivo per topi di laboratorio

Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. Previous ...

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

Analisi apprendimento spaziale e comportamento prosociale nei topi utilizzando il labirinto di Barnes e paradigmi Damsel in Distress

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...

A Standardized Protocol for Preference Testing to Assess Fish Welfare

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the addition of physical objects or conspecifics in the housing environment) is often a way to give captive animals the opportunity to choose who or what they interact with and how they spend their time. A fundamental component of the aquatic environment that is often overlooked in captivity, however, is the ability for the animal to choose to engage in physical exercise. For many animals, including fish, exercise is an important aspect of their life history, and is known to have many health benefits, including positive changes in the brain and behavior. Here we present a method for assessing habitat preferences in captive animals. The protocol could easily be adapted to look at a variety of environmental factors (e.g., gravel versus sand as a substrate, plastic plants versus live plants, low flow versus high flow of water) in different aquatic species, or for use with terrestrial species. Statistical assessment of preference is carried out using Jacob&#39;s preference index, which ranks the habitats from -1 (avoidance) to +1 (most preferred). With this information, it can be determined what the animal wants from a welfare perspective, including their preferred location.

A fundamental aspect of assessing the welfare of animals in captivity is to ask whether the animals have what they want. Here, we present a protocol to determine housing preference in the zebrafish (Danio rerio) with respect to the presence/absence of environmental enrichment and access to flowing of water.

Un protocollo standardizzato per i test delle preferenze per valutare il benessere dei pesci

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the ...

The Innovation Arena: A Method for Comparing Innovative Problem-Solving Across Groups

Problem-solving tasks are commonly used to investigate technical, innovative behavior but a comparison of this ability across a broad range of species is a challenging undertaking. Specific predispositions, such as the morphological toolkit of a species or exploration techniques, can substantially influence performance in such tasks, which makes direct comparisons difficult. The method presented here was developed to be more robust with regard to such species-specific differences: the Innovation Arena presents 20 different problem-solving tasks. All tasks are presented simultaneously. Subjects are confronted with the apparatus repeatedly, which allows a measurement of the emergence of innovations over time - an important next step for investigating how animals can adapt to changing environmental conditions through innovative behavior.
Each individual was tested with the apparatus until it ceased to discover solutions. After testing was concluded, we analyzed the video recordings and coded successful retrieval of rewards and multiple apparatus-directed behaviors. The latter were analyzed using a Principal Component Analysis and the resulting components were then included in a Generalized Linear Mixed Model together with session number and the group comparison of interest to predict the probability of success.
We used this approach in a first study to target the question of whether long-term captivity influences the problem-solving ability of a parrot species known for its innovative behavior: the Goffin&#180;s cockatoo. We found an effect in degree of motivation but no difference in the problem-solving ability between short- and long-term captive groups.

The Innovation Arena is a novel comparative method for studying technical innovation rate per time unit in animals. It is composed of 20 different problem-solving tasks, which are presented simultaneously. Innovations can be carried out freely and the setup is robust with regard to predispositions on an individual, population or species level.

L'arena dell'innovazione: un metodo per confrontare la risoluzione di problemi innovativi tra i gruppi

Problem-solving tasks are commonly used to investigate technical, innovative behavior but a comparison of this ability across a broad range of species is ...

A Behavioral Task Modeling 'Everyday Memory' in an Event Arena to Foster Allocentric Representations for Rodents

The event arena provides an optimal platform to investigate learning and memory. The appetitive everyday memory task described in this paper provides a robust protocol for the investigation of episodic and spatial memory in rodents, which specifically fosters allocentric memory representation. Rats are trained to find and dig for food during the encoding phase and, after a time delay, rats are given a choice to find the reward food pellet in the correct location. There are two key elements that promote the use of an allocentric strategy in this protocol: 1) rats start from different start locations within and between sessions, 2) a stable home-base is deployed where rats have to carry their food to eat. By means of these modifications, we effectively encourage the rodents to use allocentric spatial representations to perform the task. In addition, the task provides a good paradigm for within-subject experimental design and allows experimenters to manipulate different conditions to reduce variability. Used in conjunction with behavioral and physiological techniques, the resulting rodent model provides an effective test-bed for future research into memory formation and retention.

<meta charset="utf8"></head><body>Un compito comportamentale che modella la "memoria quotidiana" in un'arena di eventi per promuovere rappresentazioni allocentriche per i roditori

The goal of this optimized 'everyday memory' protocol in an event arena was to employ a stable home-base that encourages the use of allocentric spatial representations. This animal model provides an effective test-bed for future research into the formation and retention of event memories using behavioral and physiological techniques.

Un compito comportamentale che modella la "memoria quotidiana" in un'arena di eventi per promuovere rappresentazioni allocentriche per i roditori

The event arena provides an optimal platform to investigate learning and memory. The appetitive everyday memory task described in this paper provides a ...

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Un compito comportamentale che modella la "memoria quotidiana" in un'arena di eventi per promuovere rappresentazioni allocentriche per i roditori

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