Beatrice Tscherning Olesen, Karoline Jakobsen, Tine Bj&#248;rnholt Gustafsen, Helena C&#233;line Kj&#230;r Byriel and Kira Sonnichsen Graahede are acknowledged for their great efforts, feedback and assistance in designing the checklist and using it in their daily work. Medical illustrator Ken Kragsfeldt is acknowledged for his professional help illustrating figure 3.

The pigs used to validate the protocol were part of studies approved by The Animal Experimental Council - a national governmental institution evaluating and approving all animal experiments in Denmark. The Council does the ethical review similar to an IACUC.
1. Arrival at the research facility
<ol>
	<li>First, wash the pigs in a designated area to allow a thorough visual inspection. Use an exclusion list to exclude pigs with wounds, ear bites, abscesses, lameness etc., so only animals that a...

<ol>
	<li>Burch, R. L., Russell, W. M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The principles of humane experimental technique. , (1959).</li><li>The 3Rs. National Centre for the Replacement Available from: <a target="_blank" href="https://www.nc3rs.org.uk/the-3rs">https://www.nc3rs.org.uk/the-3rs</a> (2021)</li><li>Gouveia, K., Hurst, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimising+reliability+of+mouse+performance+in+behavioural+testing:+the+major+role+of+non-aversive+handling.">Optimising reliability of mouse performance in behavioural testing: the major role of non-aversive handling.</a> Scientific Reports. 7, (2017).</li><li>Laule, G. E., Thurston, R. H., Alford, P. L., Bloomsmith, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Training+to+reliably+obtain+blood+and+urine+samples+from+a+diabetic+chimpanzee+(Pan+troglodytes).">Training to reliably obtain blood and urine samples from a diabetic chimpanzee (Pan troglodytes).</a> Zoo Biology. 15 (6), 587-591 (1996).</li><li>Mills, D., Ledger, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+oral+selegiline+hydrochloride+on+learning+and+training+in+the+dog:+A+psychobiological+interpretation.">The effects of oral selegiline hydrochloride on learning and training in the dog: A psychobiological interpretation.</a> Progress in Neuro-Psychopharmacology &amp; Biological Psychiatry. 25 (8), 1597-1613 (2001).</li><li>Arluke, A., Sanders, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Regarding Animals. , (1996).</li><li>Leidinger, C., Herrmann, F., Thone-Reineke, C., Baumgart, N., Baumgart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introducing+Clicker+Training+as+a+Cognitive+Enrichment+for+Laboratory+Mice.">Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice.</a> Journal of Visualized Experiments. (121), e55415 (2017).</li><li>Leidinger, C. S., Kaiser, N., Baumgart, N., Baumgart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+Clicker+Training+and+Social+Observation+to+Teach+Rats+to+Voluntarily+Change+Cages.">Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages.</a> Journal of Visualized Experiments. (140), e58511 (2018).</li><li>Sorensen, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Never+wrestle+with+a+pig.">Never wrestle with a pig.</a> Lab Animal. 44 (2), 159-161 (2010).</li><li>. 2019 report on the statistics on the use of animals for scientific purposes in the Member States of the European Union in 2015-2017 Available from: <a target="_blank" href="https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:52020DC0016&amp;from=EN">https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:52020DC0016&amp;from=EN</a> (2020)</li><li>Louise, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target+training+pigs+within+an+isolation+unit+(a+pilot+study.">Target training pigs within an isolation unit (a pilot study.</a> Animal Technology and Welfare. 17 (1), 46-47 (2018).</li><li>Ryden, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nursing+and+training+of+pigs+used+in+renal+transplantation+studies.">Nursing and training of pigs used in renal transplantation studies.</a> Laboratory Animals. , (2019).</li><li>Smith, A. C., Swindle, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+swine+for+the+laboratory.">Preparation of swine for the laboratory.</a> ILAR Journal. 47 (4), 358-363 (2006).</li><li>Swindle, M. M., Smith, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Swine in the Laboratory: Surgery, Anesthesia, Imaging, and Experimental Techniques, Third Edition. , (2016).</li><li>Obernier, J. A., Baldwin, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishing+an+appropriate+period+of+acclimatization+following+transportation+of+laboratory+animals.">Establishing an appropriate period of acclimatization following transportation of laboratory animals.</a> ILAR Journal. 47 (4), 364-369 (2006).</li><li>Kaiser, A. R., Johnson, A. K., Ross, J. W., Selsby, J. T., Stalder, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Independent+Study+490A:+Positive+Reinforcement+Training+Piglets+to+Stand+in+a+Container+and+Follow+a+Human.">Independent Study 490A: Positive Reinforcement Training Piglets to Stand in a Container and Follow a Human.</a> Iowa State University Animal Industry Report. 11 (1), (2014).</li><li>S&#248;rensen, D. B., Dragsted, N., Glerup, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positive+reinforcement+training+in+large+experimental+animals.">Positive reinforcement training in large experimental animals.</a> Altex. 28, 250 (2011).</li><li>Lawrence, A. B., Vigors, B., Sandoe, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+Is+so+Positive+about+Positive+Animal+Welfare?-A+Critical+Review+of+the+Literature.">What Is so Positive about Positive Animal Welfare?-A Critical Review of the Literature.</a> Animals. 9 (10), (2019).</li><li>Jirkof, P., Rudeck, J., Lewejohann, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+Affective+State+in+Laboratory+Rodents+to+Promote+Animal+Welfare-What+Is+the+Progress+in+Applied+Refinement+Research.">Assessing Affective State in Laboratory Rodents to Promote Animal Welfare-What Is the Progress in Applied Refinement Research.</a> Animals. 9 (12), (2019).</li><li>Herskin, M. S., Bundgaard, C. J., Ottesen, J. L., S&#248;rensen, D. B., Marchant-Forde, J. N., S&#248;rensen, D. B., Cloutier, S., Gaskill, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Animal-centric care and management. , 173-186 (2020).</li><li>What is a Culture of Care. Available from: <a target="_blank" href="https://norecopa.no/more-resources/culture-of-care">https://norecopa.no/more-resources/culture-of-care</a> (2020)</li><li>Bertelsen, T. H., S&#248;rensen, D. B., Cloutier, S., Gaskill, B. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=in+.+eds+Dorte+Bratbo+S&#248;rensen.">in . eds Dorte Bratbo S&#248;rensen.</a> Animal-centric care and management. , 15-29 (2020).</li><li>Weichbrod, R. H., Thompson, G. A., Norton, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Management of Animal Care and Use Programs in Research, Education, and Testing. 2 edn. , (2017).</li><li>SPF-DANARK. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SPF+status+designations.">SPF status designations.</a> SPF-DANARK. , (2021).</li><li>Berset Convenor, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Federation+of+European+Laboratory+Animal+Science+Associations+recommendations+of+best+practices+for+the+health+management+of+ruminants+and+pigs+used+for+scientific+and+educational+purposes.">Federation of European Laboratory Animal Science Associations recommendations of best practices for the health management of ruminants and pigs used for scientific and educational purposes.</a> Lab Animal. , (2020).</li><li>Rehbinder, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FELASA+recommendations+for+the+health+monitoring+of+breeding+colonies+and+experimental+units+of+cats,+dogs+and+pigs.+Report+of+the+Federation+of+European+Laboratory+Animal+Science+Associations+(FELASA)+Working+Group+on+Animal+Health.">FELASA recommendations for the health monitoring of breeding colonies and experimental units of cats, dogs and pigs. 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The implementation of a protocol that includes part one, a transition phase checklist and part two, a consecutive clicker training program is a confirmed success. Introduction and use of a detailed checklist have provided a way to refine the introduction of pigs to the research facility. The structured transition phase furthermore creates a better foundation to train the pigs prior to inclusion in research projects. It has been a positive surprise to notice that the caretakers&#39; commitment to the work increases and th...

The principles of the 3Rs (Replacement, Reduction and Refinement) discussed by Russell and Burch in 1959, form today&#39;s foundation for high ethical standards concerning the use of animals for experimental purposes. In biomedical research facilities engaged in preclinical laboratory animal studies, great importance must be adhered to the refinement principle. &#34;Refinement means any decrease in the incidence or severity of inhumane procedures applied to those animals which still have to be used&#34;1.
A contemporary definition additionally states that refinement is &#34;advancing research into animal welfare by exploiting the latest in vivo technologies and by improving understanding of the impact of welfare on scientific outcomes&#34;2. This implies that not only are today&#39;s refinement initiatives important for the welfare of the animal per se, but that they are also important to research quality as scientific outcomes will benefit from these initiatives 3.
One refinement initiative to consider is to socialize and train experimental animals. They may be trained to perform a certain task e.g., step onto a scale or behave calmly while blood samples are drawn. Some species may be more naturally disposed to training than others and this impacts the tradition for training laboratory animal species. Traditionally, training of dogs has been performed for centuries e.g., in order to use the dog in hunting. These historic traditions have most certainly made the training of dogs in the biomedical research setting more straightforward. Together with non-human primates, training of laboratory dogs has been discussed and undertaken for quite some time4,5.
Common to both of these species is that they rank high on the &#34;socio-zoological scale&#34;. This scale ranks animals according to their place and use in human society 6. Ranking high also carries with it a large public demand to treat the animals as humanely as possible and limit their stress and anxiety level. Rodents, in contrast to dogs and non-human primates, are placed in the lower end of the socio-zoological scale. They have not received equivalent public attention and care. Historically they have been treated less well - presumably also due to their ability to spread life-threatening disease. However, during the last few years the training of rodents has been implemented in preclinical research settings and hereof related scientific benefits have not gone unnoticed. Both rats and mice are interestingly quite easily trained and the training also limits their stress-response and thus strengthens scientific outcomes7,8.
Reporting on how to socialize and train pigs in a laboratory animal setting is to our knowledge scarce9. On the socio-zoological scale, pigs are placed in-between dogs and rodents. We recognize the species primarily as a food resource, but nevertheless quite a large number of pigs are also used in biomedical research. In the EU alone, approximately 75,000 pigs were used annually for scientific purposes in 2015-2017 10. This number encompasses both farm pigs and miniature pigs. There is an increasing focus on refining the lives of these animals by training as a measure to comply with the 3Rs. Both farm pigs and miniature pigs can indeed be trained 11,12, but their starting point in terms of socialization varies. Miniature pigs are bred for research and socialized during their early life span. They are bred to have a calm temper. Farm pigs are bred for consumption and often not socialized at all. Finisher pigs for meat production in Denmark are typically housed in high density groups of 15-25 pigs where they have to compete to earn a position in the herd&#39;s hierarchy. Their temper reflects that ability. Often, they are housed in stable facilities with a partially slated floor to limit the workload of mucking out. To optimize weight gain, they are fed ad libitum through automated food and water delivering systems. Hence, their contact with farm workers is limited and they will become stressed when handled. This poses certain challenges when the animals are transferred to a research facility. Here the animal welfare, the husbandry, and the 3Rs play a key role13,14. From a research perspective, each individual animal is important to obtain&#160;the best possible research results. The welfare of the animals is also important to ensure a low inter-animal variation in regard to stress, anxiety and fear. For these reasons it is important to pay attention to the transition from a &#34;herd animal&#34; to an &#34;individual&#34;. Upon arrival, the pigs enter a new environment with new scents, new food, day-to-day contact with animal caretakers and often also new pen mates. To a pig, the shift can be very stressful and the importance of an acclimatization period of several days is acknowledged in laboratory animal science15.
By structuring the acclimatization period, we have created what we call a &#34;transition phase&#34; using a structured step-by-step checklist. The checklist ensures that each pig receives sufficient caretaker contact and learns to allow basic handling for examination and sample collection without being stressed. Continuous work can then be carried out to train the pigs using positive reinforcement training targeted to each specific research project16,17. A clicker is used to signal a food reward when the desired action is performed. Combining the steps from the transition phase with additional clicker training of the pigs gives the animals a smooth settling in at the research facility. They become calm and easy to handle and can, with little effort, be taught to stand still for e.g. blood sample collection. By the use of this type of handling protocol, old-fashioned techniques such as sedation of the animals or snares are replaced. To make sure all pigs are handled sufficiently a checklist is filled out.
It is becoming widely accepted that animals do indeed feel positive emotions and so focus when working with the animals should not solely be on the removal of negative emotions 18. The training session with the caretaker is an example of a positive emotional experience. It is quite enjoyable for the pig as it receives treats and attention. Besides the benefit of reducing handling-stress and fear related behaviors at the facility, training the animals provides an opportunity to give the animals positive emotional experiences. The adding of positive experiences may counteract some of the negative experiences that laboratory animals encounter in relation to experiments performed and will thus further increase animal welfare 19,20.
While the pigs evidently enjoy the training sessions, the sessions simultaneously offer the caretaker an opportunity to strengthen the bond to each individual pig. Being an animal caretaker, performing painful actions, such as injections of the animals, is part of the job. However, being able to train the animals and turn these actions into positive experiences is rewarding in itself. It increases the commitment of the caretakers and plays a key role in strengthening the department&#39;s culture of care21,22,23.
The pigs were housed according to the Institution&#39;s Animal Welfare Policy, which refers to national legislation on animal experimentation, housing and husbandry. The protocol can be applied to weaned pigs of any age. For the development of the transition checklist (Figure 1) a total of 40 female crossbred farm pigs (Danish Landrace-Yorkshire-Duroc) weighing 35-80 kg were used (Table 1). The pigs were allocated to a group dependent on their arrival time. The pigs were housed in the research stable facility in the period January - September 2020. All farm pigs are bred on a traditional sow farm and moved to a collaborating farm when weighing approximately 15 kg. At least two weeks before inclusion in survival studies, the pigs are transported to the university&#39;s research stable facility. During the first two weeks the transition from farm pig to research animal takes place. The protocol is composed of non-harmful and positive actions and hence may be initiated immediately as part of the acclimatization period.
The pigs were housed one by one in pens measuring 3.4 - 6.8 m2 on solid concrete floors and have access to water via an automatic water supply. There is a feeding trough for each pig and at least one section of the separation to the neighboring pens allows snout contact. When pigs are scheduled to receive surgical interventions or have equipment implanted, they are housed alone to avoid pen mates licking and biting in wounds and pulling implants out. The bedding material is straw and the animals are further enriched with hay and different activity toys e.g. rope, balls, buckets, chew sticks made from plastic (Figure 2).
The animals come from a herd included in the national health-monitoring program for farm pigs, which means they are screened for seven pathogens24 that can affect pigs in a production setting. Biannual random checks are performed for the research herd based on FELASA recommendations25,26, in addition to samples from pigs that warrant diagnostics due to unexpected research results. Some of the pigs used for this project were seropositive for porcine reproductive and respiratory syndrome; however, none of the pigs showed any clinical symptoms corresponding to infection. All samples evaluated using polymerase chain reaction were negative, corroborating that the pigs were in a healthy condition.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Wrap, e-power, bl&#229;, 7,5cm</td>
			<td>E-vet A/S, Ole R&#248;mers Vej 26 A, DK-6100 Haderslev</td>
			<td>203165B</td>
			<td>Adhesive wrap used to wrap around the end of a stick to make a target stick. It is an advantage that it can be changed as it will get dirty quite quickly.</td>
		</tr>
		<tr>
			<td>Apples</td>
			<td>Varius</td>
			<td>Uknown</td>
			<td>Used as treats</td>
		</tr>
		<tr>
			<td>Klikker (Clicker)</td>
			<td>E-vet A/S, Ole R&#248;mers Vej 26 A, DK-6100 Haderslev</td>
			<td>TX2289</td>
			<td>Clicker used in training</td>
		</tr>
	</tbody>
</table>

transition of farm pigs to research pigs using a designated checklist followed by initiation of clicker training - a refinement initiative

Implementation of 3R initiatives is important. To refine the life of laboratory pigs, we focused on the time period prior to inclusion of animals in experiments. We improved the checklist during the development. By using the checklist each individual pig&#39;s progress was documented. This aids the caretakers so they remain focused on the actions necessary to socialize the animals and limit the animal&#39;s stress and anxiety level. During this phase, the pigs become calm and ready to commence additional clicker training to further limit stress during the experimental period. The pigs will readily allow basic outcome measures to be collected without sedation or fixation.
The protocol is divided in two. First the transition phase that begins as the pigs arrive at the research facility. Introducing treats and gradually closer contact to the caretakers are the first steps. Then, the caretaker introduces actions to be able to touch the head and hind, to mimic e.g. manipulating the ear to take a blood sample or the hind to obtain a rectal temperature. A strong foundation is established in the animal-caretaker relationship and this swiftly minimizes the stress the pigs express after arrival. Secondly, the positive reinforcement training commences to teach the animal how to carry out more complex tasks without becoming stressed. A clicker is introduced as a secondary reinforcer.
Time, structure, education and thorough communication seem to be the most important criteria to succeed in the application of this protocol. In addition to having calm and compliant pigs, the caretakers report that they enjoy the focus they are allowed to give each individual pig, adding to their overall job satisfaction. Having caretakers with a strong commitment, and a focus on the transition progress and welfare of each pig, additionally strengthens the department&#39;s Culture of Care.

Applying the transition phase as described in the protocol&#39;s part one allows for a smooth introduction of farm pigs to the research facility. Having calm pigs to work with eases daily chores and enables minor procedures and sample taking to be performed without anesthesia or fixation.
To follow the progress of pigs during the transition phase a checklist (Checklist 1) was developed to mark when individual pigs reach defined steps in their transition. The pigs are scheduled to enter researc...

Watch this Scientific Journal Video about Transition of Farm Pigs to Research Pigs using a Designated Checklist followed by Initiation of Clicker Training - a Refinement Initiative at JoVE.com

Transition of Farm Pigs to Research Pigs using a Designated Checklist followed by Initiation of Clicker Training - a Refinement Initiative

Refinement of porcine studies is achieved by introducing a standardized checklist and positive reinforcement training using a clicker. This work supports the collection of samples and the conduct of daily chores related to the animals.

Implementation of 3R initiatives is important. To refine the life of laboratory pigs, we focused on the time period prior to inclusion of animals in ...

This video will introduce the protocol used to transition pigs from primarily conventional farming to research pigs, using the combination of treats, socialization, and positive reinforcement training. As the pigs arrive they are washed and sorted into groups according to the projects they are to be enrolled in. On the arrival day the pigs are left to settle, and the staff limits moving about in the staples.On the day following arrival the protocol begins. The pig is in its home pen during the transition phase as this is what the pig is familiar with, and therefore gives the best basis of progression when the goal is to make the pig calm in the presence of the caretaker. The first step is introducing treats.Farmed pigs have never had any other food types than their pig feed so they need to be taught what a treat is. We use apple pieces as a standard type of treat. The first step is to simply toss pieces of apple into the pen and let the pig find it.In the beginning the pigs may pay no notice to the apples, but usually within a day or two they understand that a piece of apple is nice, and something to look forward to. In the protocol a plus is given when the pig eats pieces of apple within two minutes, and a minus is given if the pig has not found the apples, or won't eat it during those two minutes. The caretaker is positioned just outside the pen to watch the pig, and also to get the pig used to the close presence of the caretaker.From day three the caretaker enters the pen during feeding. The idea behind this is to take advantage of the pig strong motivation to eat, and combine that with the close presence of the caretaker. The caretaker intercept pen, pours the feed into the trough, and then sits down next to the pig.If the pig comes to eat the feed the caretaker stays. If the pig refuses to eat while the caretaker sits next to the trough, the caretaker stays for two minutes, and then leaves the pen. When the pig accepts the close presence of the caretaker during feeding the caretaker slowly extends an arm to touch the pigs back.If the pig backs away from the touch a minus is recorded in the protocol. If the pig accepts the touch a plus is recorded. Once touching is accepted it is slowly increased.Both in the area touched, and the speed of the movements. Touching of the head, and ears, and hind, and tail, is useful for most projects, so they are scored in the protocol. From the sitting position the hand is moved from the back to the head of the pig, and then moved around to also touch the ears.When this is tolerated more manipulation of the ears is the next step. When the pig accepts being touched on the head, and having its ears touched and manipulated, a plus is noted in the protocol until then a minus is recorded. Touching the hind and tail is performed by moving the hand backwards on the pig.In this step the caretaker also rises slowly to be able to move around the pig for better positioning. Standing up and bending towards the pig is more frightening to the pig, so all the movements should be performed slowly. When the pig accepts being touched on the hind, and having the tail touched, and manipulated during a two minute session, a plus is recorded in the protocol.If during two minutes the pig does not accept this a minus is recorded. In addition to entering the pen twice daily during feeding, the caretaker also enters the pen during the day to further accelerate the bonding process between caretaker and pig. During these sessions the pigs are offered treats by hand.The caretaker sits just inside the pen, and offers a piece of apple to the pig with an outstretched arm. If the pig eats the apple during the two minute timeframe a plus is recorded in the checklist. If it does not eat the apple from the hand a minus is recorded.The final step in the transition phase is for the pig to accept being touched by one caretaker while a second caretaker gives the pig treats. This is harder for the pig to accept as it is more aware of its surroundings, and two people are now in close proximity. On completion of the transition phase the pigs will allow collection of basic outcome measures like taking a temperature, manipulating the ears to collect a blood sample, or attaching a bag to collect a urine sample.Following the completion of the transition phase the pigs are ready to start positive reinforcement training. A clicker is used to signal the coming reward. The use of a clicker compared to a voice signal enables different caretakers to train the same animals.When the clicker training is initiated the pigs are taken outside their home pen as more space is needed to have the pig move freely about. The free space between rows of pens, or designated room can be used. Shortly after the introduction of the clicker a target stick is introduced.A target stick is simply a stick with a mark area at one end. We have made one ourselves using a bamboo stick, and adhesive tape. That makes it easy to change the tape as it becomes dirty from contact with the snouts of the pigs.The pig is allowed to roam about but eventually it will make contact with the caretaker, and the caretaker clicks, and gives the pig a reward in one motion. Multiple clicks are in quick succession. Every click is followed by a reward.The pig needs to understand that when it hears a click a reward will follow immediately. A few sessions should be used to make sure that the pig understands that a click means a treat, as it is important to be able to give the signal to the pig quickly to be able to reward a specific behavior. When the link between the click and the reward is well understood by the pig, the target stick can be introduced.The target end is held in front of the pig, and at some point the pig will touch it out of curiosity. At the very second the snout touches the target click and reward. The target is positioned again, and the task is repeated a number of times.Then gradually the target is moved farther away from the pig to make the pig move to touch it. Within a few sessions the pig will have learned to move around following the target stick. In the beginning the pig will break contact from the target if distracted by new objects, other pigs, or sounds.But with consistent training the pig will be able to follow the target stick closely even when disturbances occur. When the pig can follow the target stick the stick can be used to move the pig around the facility. At our department it's used to guide the pigs into transport crates, or to move the pigs to our floor scale, and back to their home pen.In conclusion, the implementation of a structured transition phase has highlighted how fast and smoothly the pigs can adjust to the research facility, and that they are notably less stressed when they enter experiments. Reduction of the pig stress level is a key part of our 3R strategy to improve animal welfare and science. The dedication of our caretakers has increased as well, and this contributes to the department's culture of care, and the collaboration between scientists and caretakers.

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JoVE Journal

Behavior

4.9K visualizações.  Aarhus University. Este vídeo introduzirá o protocolo usado para a transição de suínos da agricultura convencional para a pesquisa de suínos, usando a combinação de guloseimas, socialização e treinamento de reforço positivo. À medida que os porcos chegam, eles são lavados e classificados em grupos de acordo com os projetos em que devem ser inscritos. No dia da chegada, os porcos são deixados para se estabelecer, e os limites dos funcionários se movem nos grampos.No dia seguinte à chegada ...