The authors have no acknowledgements.

Ethics statement: All procedures were approved by the Animal Ethics Committee of the University of Sydney and conformed to National Health and Medical Research Council of Australia guidelines. Procedures were performed on C57/BL6J mice which were reared at the University of Sydney Bosch Rodent Facility. All mice were housed in a single adequately-ventilated room in 21 &#176;C ambient temperature on a 12 hr light-dark cycle with lights on at 0600 hr in individually ventilated cages with ad libitum access to dry food and water. Late-pregnancy females were randomly assigned to standard or environmentally enriched housing conditions.
1. Housing (Enrichment Levels):
<ol>
	<li>Obtain 4 late-pregnancy adult female mice. Randomly assign 2 to the standard condition and place each of them into a clean standard mouse cage (overall dimensions 391 x 199 x 160 mm) containing one red mouse igloo. For the enriched condition, place the remaining 2 mice into a single clean rat-sized cage (overall dimensions 462 x 403 x 404 mm).</li>
	<li>Into the enrichment cage, place a variety of objects designed to increase sensory and motor stimulation (e.g. running wheels, visual stimuli, scented cotton balls, Velcro strips).</li>
	<li>Every 2 to 3 days move these objects about within the cage; refresh any that have been destroyed.</li>
	<li>Upon weaning at 21 days postnatal, sex animals and place into male-female segregated housing consistent with the environmental condition they were raised. For the enriched condition ensure that there are between 3 and 10 mice per cage with 2 and 5 mice for standard. Commence behavioral testing once animals reach adulthood (12&#8211;14 weeks of age).</li>
</ol>
2. Construction of the Puzzle Box
<ol>
	<li>Obtain 6 pieces of white acrylic (or other non-porous material): one 750 x 280 mm, two 280 x 250 mm, two 750 x 250 mm, and one 150 x 280 mm (see Figure 1).</li>
	<li>Obtain one piece of black acrylic 280 x 250 mm, with a 40 x 40 mm square opening cut into one side of this piece (see Figure 1).</li>
	<li>Assemble the Puzzle Box as follows: use the 750 x 280 mm piece as the bottom of the box, use 280 x 250 mm pieces as the ends of the box, and the 750 x 250 mm pieces as the sides of the box.</li>
	<li>Measure 150 mm into the box from one end, and place the black piece of acrylic across the box so that it splits it into two compartments (one large, one small) with the opening flush with the bottom of the box.</li>
	<li>Take the 150 x 280 mm piece of white acrylic and place it atop the smaller compartment of the box making sure it covers this area completely, providing a dark &#8220;goal-box&#8221; chamber. Affix this piece of acrylic to the body of the main box by hinges, or leave free to be completely removed during behavioral testing.</li>
	<li>Take 3 pieces of acrylic (three 4 x 120 mm) and join to make a &#8220;u-shaped&#8221; channel.</li>
</ol>
<img alt="Figure 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225fig1highres.jpg" width="500" /> 
Figure 1: Schematic diagram of the Puzzle Box task. The Puzzle Box is an acrylic box consisting of an open-field area (600 x 280 mm) and a sheltered goal-box area (150 x 280 mm), measuring 750 x 280 mm in all. A 40 x 40 mm opening within the partition (grey) separating the two areas allows animals to access to the covered goal-box area from the open field. This opening is blocked with obstructions that are increasingly difficult to remove as testing progresses. Animals undergo a five-day protocol, consisting of four obstruction conditions with three trials for each condition.
3. Running of the Puzzle Box Task
<ol>
	<li>Thoroughly clean the Puzzle Box with 70% alcohol. Repeat this step between each animal tested.</li>
	<li>Place a clean red mouse igloo within the goal-box area of the Puzzle Box, and replace the lid on the goal-box.</li>
	<li>If there is an obstruction condition being tested, place the obstruction within the doorway of the goal-box (see Figure 2).</li>
	<li>Place the mouse being tested into the open-field section of the Puzzle Box, oriented towards the goal-box, and at the end furthest from the goal-box.</li>
	<li>Record the time taken for all four paws of the animal to enter the goal-box section of the Puzzle Box.</li>
	<li>If an animal does not enter the goal-box, terminate the trial once the set time limit is reached (see Table 1).</li>
	<li>Once a trial is finished, remove the animal from the Puzzle Box and place it into a separate holding cage until the next trial begins. Keep a 60 to 180 sec gap between trials for each animal.</li>
	<li>For each animal, perform three trials per day for five consecutive days of testing, with four obstruction conditions and three trials of each condition. The third trial of a given obstruction condition should&#160;always administered on a subsequent day (see Table 1).</li>
</ol>
<img alt="Table 1" fo:content-width="5in" src="/files/ftp_upload/52225/52225table1.jpg" width="500" /> 
Table 1: Scheme of the Puzzle Box task. The Puzzle Box task is run for five days, and consists of four obstruction conditions. There are three trials on each of the first four days, and one on the fifth day. Each obstruction condition has three trials; the first two on one day, and the third the day immediately following. The first trial of an obstruction condition aims to test native problem-solving ability, the second trial examines task acquisition and reinforcement, and the third trial is used as an assay for solution retention and recall.
<img alt="Figure 2" fo:content-width="6in" src="/files/ftp_upload/52225/52225fig2highres.jpg" width="600" /> 
Figure 2: Obstruction conditions within the Puzzle Box task. Schematic diagrams&#160;of the Puzzle Box arena and the obstruction conditions used within this study. (A) Condition 0 (C0), with no obstruction present within the doorway between the open-field and goal-box areas. (B) Condition 1 (C1), with a &#8220;u shaped&#8221; channel present within the doorway between the open-field and goal-box areas. (C) Condition 2 (C2), where the channel is filled with clean bedding material. (D) Condition 3 (C3), with a &#8220;tissue plug&#8221; present within the doorway between the open-field and goal-box areas. (E) Condition 4 (C4) with a &#8220;foam plug&#8221; present within the doorway between the open-field and goal-box areas. Dimensions of the arena are as per listed in Methods and Figure 1.
4. Interpretation of Data
<ol>
	<li>Represent data as either time taken to complete the trial (including null trials where animals did not complete the task within the specified time limit) or as the number of null trials.</li>
	<li>Use a repeated measures ANOVA to assess the effect of housing condition upon performance within the Puzzle Box, with obstruction type/condition (C) and task number (T) within-subjects factors and enrichment level (standard versus enriched) as between-subjects factor.</li>
</ol>

The authors declare they have no competing financial interests.

The data presented demonstrate that the Puzzle Box can be used effectively to assess the impact of EE. Mice raised in enriched environments consistently took significantly less time to solve obstruction puzzles within this behavioral assay than did animals raised within standard laboratory conditions. Moreover, this difference was most prominent in the first trial for three of the four conditions tested, suggesting EE has a greater influence on an animal&#8217;s native problem-solving ability, relative to their capacity ...

Environmental enrichment (EE) may be defined as surroundings that provide animals with increased opportunity for social interaction, motor activity, and greater sensory stimulation than usually experienced in a standard laboratory environment.EE has been shown to consistently affect the behavior of animals, bringing about changes such as reduction of stress and anxiety-related activity 8-10, improved performance in learning and memory tasks 8,11, early onset of motor coordination and exploratory activity 11, changes in maternal care 8 as well as resistance to addictive substances 12-15. Further, EE has been revealed to ameliorate the effects of neurodegenerative disorders, delaying the onset and decreasing the severity of symptoms in animal models of Huntington&#8217;s 1-4,16, Parkinson&#8217;s 17 and Alzheimer&#8217;s disease 18.
These changes correlate with the anatomical and molecular alterations EE is known to induce throughout the brain. Animals raised in enriched environments from early stages of development show a myriad of neural changes, including increased brain weight and cortical thickness, dendritic branching 9,2-22 and synaptic density 23. EE can alter both the level and timing of growth factor expression 9,24-30, which has been shown to contribute to accelerated development of sensory 25,26,28,29, mnemonic 30, as well as motor circuits 31,32.
Previous work has revealed at times contradictory findings when investigating the impact of EE, without taking into account the different types of animals and environments used within individual studies 9,24,27,30. Currently, there is no consistent and simple behavioral task that can be used to measure the effectiveness of various EE paradigms in different strains and species of animals.
The Puzzle Box task was designed as a simple test to determine an animal&#8217;s native problem solving ability 7. Animals placed in the open area are required to remove obstructing materials situated within a small opening in order to access a covered region/shelter. Each subject is given three trials with the same obstruction in order to assess three different cognitive attributes. The first trial yields a baseline indication of inherent or native problem solving ability. The second trial, run on the same day, gives some indication of the animal&#8217;s ability to improve upon and thus reinforce strategies for removing the specific obstruction. The third trial, conducted on the following day, provides insight into the ability of the subject to retain and recall the learned solution to the task.
The motivation for solving these &#8220;obstruction puzzles&#8221; by the animals can be varied, potentially evoking an innate desire to avoid open fields and seek shelter, as well as an inherent drive to explore their surroundings 6,7. The multitude of potential behavioral drivers underlying the desire to solve the Puzzle Box suggests that various areas of the brain are involved in mediating task performance. Previous work has shown that in murine models of schizophrenia, the prefrontal cortex as well as the hippocampus are involved in the acquisition of this task 5. A lesion study in rats has also revealed a large number of brain regions involved in Puzzle Box performance, including various thalamic nuclei, the hypothalamus, the cerebellum, and limbic structures. Together, these findings indicate that engaging in this problem solving task involves a host of neural structures associated with cognitive function.
The Puzzle Box has been used successfully to assess the problem solving ability of mice, as well as cognitive deficits exhibited by murine models of schizophrenia 5-7. Performance on the task has been shown to be highly consistent, and correlate well with outcomes of other cognitive behavioral tests 6. The goal of this work was thus to adapt the Puzzle Box task to become a simple and reliable means of determining the effectiveness of EE.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Black Acrylic Board</td>
			<td></td>
			<td></td>
			<td>750 x 280 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>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>280 x 250 mm with a 40 x 40 mm &#34;door&#34; cut into one side</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></td>
			<td></td>
			<td></td>
			<td>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&#160;</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&#160;</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.

The results described here are a representative sample, with data taken from several cohorts consisting of different litters. All behavioral testing was conducted between 0700 and 1100 hr, with randomized testing order of animals within a cohort. Animals raised in an enriched environment (n = 14, 7 female and 7 male) took significantly less time to solve the obstruction tasks within the Puzzle Box than those raised within a standard environment (n = 15, 7 female and 8 male) (see Figure 3) (Repeated measu...

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.

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

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Research

JoVE Journal

Behavior

11.5K 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 Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

Assessing Forelimb Function after Unilateral Cervical SCI using Novel Tasks: Limb Step-alternation, Postural Instability and Pasta Handling

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority of the spinal cord injuries in humans occur at the cervical levels. Therefore, developing cervical injury models and developing relevant and sensitive behavioral tests is of great importance. Here we describe the use of a newly developed forelimb step-alternation test after cervical spinal cord injury in rats. In addition, we describe two behavioral tests that have not been used after spinal cord injury: a postural instability test (PIT), and a pasta-handling test. All three behavioral tests are highly sensitive to injury and are easy to use. Therefore, we feel that these behavioral tests can be instrumental in investigating therapeutic strategies after cSCI.

Three new behavioral tests (forelimb step-alternation, postural instability test, pasta handling test) for evaluating forelimb function after cervical spinal cord injury in rodents are described.

Cervical spinal cord injury (cSCI) can cause devastating neurological deficits, including impairment or loss of upper limb and hand function. A majority ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

Assessing the Multiple Dimensions of Engagement to Characterize Learning: A Neurophysiological Perspective

In a recent theoretical synthesis on the concept of engagement, Fredricks, Blumenfeld and Paris1 defined engagement by its multiple dimensions: behavioral, emotional and cognitive. They observed that individual types of engagement had not been studied in conjunction, and little information was available about interactions or synergy between the dimensions; consequently, more studies would contribute to creating finely tuned teaching interventions. Benefiting from the recent technological advances in neurosciences, this paper presents a recently developed methodology to gather and synchronize data on multidimensional engagement during learning tasks. The technique involves the collection of (a) electroencephalography, (b) electrodermal, (c) eye-tracking, and (d) facial emotion recognition data on four different computers. This led to synchronization issues for data collected from multiple sources. Post synchronization in specialized integration software gives researchers a better understanding of the dynamics between the multiple dimensions of engagement. For curriculum developers, these data could provide informed guidelines for achieving better instruction/learning efficiency. This technique also opens up possibilities in the field of brain-computer interactions, where adaptive learning or assessment environments could be developed.

This paper aims to describe the techniques involved in the collection and synchronization of the multiple dimensions (behavioral, affective and cognitive) of learners&#8217; engagement during a task.

In a recent theoretical synthesis on the concept of engagement, Fredricks, Blumenfeld and Paris1 defined engagement by its multiple dimensions: ...

Errors as a Means of Reducing Impulsive Food Choice

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ultimately, to reduced life expectancy. The capacity to refrain from automatic responses is usually high in situations in which making errors is highly likely. The protocol described here aims at reducing imprudent preference in women during hypothetical intertemporal choices about appetitive food by associating it with errors. First, participants undergo an error task where two different edible stimuli are associated with two different error likelihoods (high and low). Second, they make intertemporal choices about the two edible stimuli, separately. As a result, this method decreases the discount rate for future amounts of the edible reward that cued higher error likelihood, selectively. This effect is under the influence of the self-reported hunger level. The present protocol demonstrates that errors, well known as motivationally salient events, can induce the recruitment of cognitive control, thus being ultimately useful in reducing impatient choices for edible commodities.

Giving in to temptation of tasty food may result in long-term overweight problems. This protocol describes how to reduce imprudent preference for edible commodities during hypothetical intertemporal choices in women by associating them with errors.

Nowadays, the increasing incidence of eating disorders due to poor self-control has given rise to increased obesity and other chronic weight problems, and ...

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

The Knob Supination Task: A Semi-automated Method for Assessing Forelimb Function in Rats

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity largely use video analysis of reaching or food manipulation. While these tasks are easy to implement and are robust across disease models, they are subjective and laborious for the experimenter. Automating traditional tasks or creating new automated tasks can make the tasks more efficient, objective, and quantitative. Since rats are less dexterous than primates, central nervous system (CNS) injury produces more subtle deficits in dexterity, however, supination is highly affected in rodents and crucial to hand function in primates. Therefore, we designed a semi-automated task that measures forelimb supination in rats. Rats are trained to reach and grasp a knob-shaped manipulandum and turn the manipulandum in supination to receive a reward. Rats can acquire the skill within 20 &plusmn; 5 days. While the early part of training is highly supervised, much of the training is done without direct supervision. The task reliably and reproducibly captures subtle deficits after injury and shows functional recovery that accurately reflects clinical recovery curves. Analysis of data is performed by specialized software through a graphical user interface that is designed to be intuitive. We also give solutions to common problems encountered during training, and show that minor corrections to behavior early in training produce reliable acquisition of supination. Thus, the knob supination task provides efficient and quantitative evaluation of a critical movement for dexterity in rats.

This manuscript describes a semi-automated task that quantifies supination in rats. Rats reach, grasp, and supinate a spherical manipulandum. The rat is rewarded with a pellet if the turn angle exceeds a criterion set by the user. This task increases throughput, sensitivity to injury, and objectivity compared to traditional tasks.

Tasks that accurately measure dexterity in animal models are critical to understand hand function. Current rat behavioral tasks that measure dexterity ...

Methods of Pairing and Pair Maintenance of New Zealand White Rabbits (Oryctolagus Cuniculus) Via Behavioral Ethogram, Monitoring, and Interventions

New Zealand White (NZW) laboratory rabbits (Oryctolagus cuniculus), as well as their ancestors the European Rabbit, are a social species that exhibit numerous benefits to being housed accordingly. Although these rabbits are innately gregarious, certain behaviors can still arise when kept in captivity, which if left unchecked, can confound research results or lead to wounding, which in extreme cases can be severe. To prevent these issues, there must be a well-structured plan for the monitoring and maintenance of paired laboratory rabbits. The purpose of this protocol is to present effective procedures for establishing newly paired NZW rabbits as well as methods for successful maintenance. Multiple methods have been tested for the creation of newly paired female rabbits from the vendor, but the most efficacious technique emphasizes capitalizing on the stress bonding from transport, urine marking, pairing in a neutral cage with no forced sharing of resources and a system of monitoring and intervention. To determine the best method of housing paired rabbits in a standard caging environment, data were collected to generate a behavioral ethogram. Behaviors were then quantified as positive, neutral or negative and were tracked across the lifespan of the pair to determine which behaviors indicated pair success or failure. With the newfound knowledge of socially housed laboratory NZW rabbit behavior, enrichment intervention was applied to alleviate aggression and prevent wounding, thus resulting in a higher percentage of successful pairs. Through several years of trialing different pairing methods, the development of the ethogram and the resulting enrichment interventions, understanding of the highly complex social constructs that dominate pair housed rabbit behavior has dramatically increased and allowed for the provision of more species-specific care and increased standards of welfare.

Methods of Pairing and Pair Maintenance of New Zealand White Rabbits Oryctolagus Cuniculus Via Behavioral Ethogram, Monitoring, and Interventions

Though European rabbits are a social species, socially housing them can be challenging. Therefore, there must be a thorough understanding of behaviors and social structures of pair-housed laboratory rabbits. Here we present a protocol to identify pairing methods, species-typical hierarchy establishment behaviors and behaviors that warrant appropriate intervention.

New Zealand White (NZW) laboratory rabbits (Oryctolagus cuniculus), as well as their ancestors the European Rabbit, are a social species that exhibit ...

Assessing the Coherence of Parents' Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with parents. This paper focuses on such a method, namely Five-Minute Speech Sample-Coherence (FMSS-Coherence). In this method, a parent is asked to speak for five uninterrupted minutes about her/his child and their relationship. The resulted narrative is coded for coherence, namely the extent to which the parent provides&#160;in the narrative&#160;a clear, consistent, multidimensional and well-supported portrayal of the child. FMSS-Coherence is based on attachment research that shows that the coherence of parents&#8217; narratives is indicative of the quality of the parent-child relationship and child adjustment. It overcomes the limitations of attachment narrative measures which are typically labor intensive. FMSS-Coherence is less nuanced than extant attachment narrative measures. Yet, studies of families from different cultural backgrounds, across different child ages and in the context of typically developing children as well as children with special needs suggest that coherence can be reliably evaluated using the FMSS procedure. Furthermore, parents&#8217; FMSS-Coherence is associated with parenting quality and children&#8217;s socio-emotional adjustment. Thus, it holds promise for researchers and practitioners who seek a relatively time- and cost-effective method for assessing the coherence of parents&#8217; narratives regarding their child.

Assessing the Coherence of Parents&#039; Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

This paper introduces a method for assessing the coherence of parents' short narratives regarding their child and their relationship using the five-minute speech sample procedure.

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with ...

Providing Meaningful Environmental Enrichment and Measuring Saliva Cortisol in Pigs Housed on Slatted Flooring

As pork is the most consumed meat worldwide, the welfare of animals in the swine industry has increasingly become a major public concern, which imposes a substantial pressure coming from customers, legislators and other stakeholders, to make management changes to improve the well-being of these animals. Several studies have demonstrated that providing environmental enrichment to pigs allows them to express their natural behavior, such as rooting and exploring, as well as nesting prior to farrowing, and is associated with reduced stress and improved production and welfare. However, many considerations should be taken into account when providing environmental enrichment, such as the type of floor, drainage and sewage systems, the pigs&#39; stage in life, the material, as well as its hanging method, height and location within the pen. The objectives of this paper are (1) to give methodologic information on how to provide a relatively simple and practical meaningful environmental enrichment for pigs which are housed on slatted floors during the different stages of their life, and (2) to demonstrate how to collect saliva samples for the measurement of cortisol concentrations, as a biomarker for acute stress. Protocols include information regarding the use of jute, cotton ropes, straw in racks, as well as chewable silicone sticks devices as environmental enrichment in pens of farrowing and lactation, weaners and finishers. In addition, the use of cotton rope for a non-invasive saliva samples collection for cortisol concentrations analysis is detailed. The protocols provided are relevant for professionals aiming to improve and monitor animal welfare, in both research and industrial swine farming.

This protocol demonstrates how to provide practical meaningful environmental enrichment for pigs which are housed on slatted flooring during the different stages of their lives, and how to collect saliva samples in a non-invasive manner for the measurement of cortisol concentrations, as a biomarker for acute stress.

As pork is the most consumed meat worldwide, the welfare of animals in the swine industry has increasingly become a major public concern, which imposes a ...