Name: providing meaningful environmental enrichment and measuring saliva cortisol in pigs housed on slatted flooring
Uploaded: 2019-09-30T14:00:00
Description: Watch this Scientific Journal Video about Providing Meaningful Environmental Enrichment and Measuring Saliva Cortisol in Pigs Housed on Slatted Flooring at JoVE.com

We thank Lahav CRO for conducting the research in their farm and to the farm&#39;s team for providing valuable technical help throughout the study. The study was funded by the Israel Chief Scientist, Ministry of Agriculture and Rural Development.

The study protocol was ethically approved by the Hebrew University&#39;s Institutional Animal Care and Use Committee (MD-16-14754-2). The study was conducted during 2017, at Lahav Animal Research Institute and the Hebrew University of Jerusalem, Israel.
1. Environmental enrichment protocol during farrowing and lactation period
NOTE: In this study, sows were housed in farrowing/lactation pens, under restraint for a limited time, from 3-5 days prior to the expected farr...

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Herein, we describe protocols detailing how to provide simple and practical environmental enrichment for pigs which are housed on slatted floor&#160;during different stages of their life, and how to non-invasively collect saliva samples for the measurement of cortisol concentrations, as a biomarker for acute stress. Straw as a bedding is considered as one of the most suitable environmental enrichment for pigs, but may be impractical on slatted floors. However, relatively inexpensive alternatives such as jute, cotton rope...

Pork is the most consumed meat worldwide, with over 1.3 billion pigs being raised and slaughtered annually1,2. In recent years, the welfare of animals in the swine industry has increasingly become a major public concern, which imposes substantial pressure, coming from customers, legislators and other stakeholders, to make management changes to improve the well-being of these animals. Several studies demonstrated that providing environmental enrichment to pigs is associated with reduced stress and improved production and welfare, as it allows the pigs to express their natural behavior, such as rooting and exploring, as well as nesting prior to farrowing3,4,5,6,7.
Pigs are considered to be intelligent animals, and have a highly inquisitive nature; thus, if a suitable environment is not provided, pigs will likely demonstrate stereotypic behavior and direct manipulative social behavior towards pen mates, which may lead to tail biting, as well as other injuries and stress8,9. Therefore, providing a meaningful environmental enrichment is advised by professionals, and in some countries even imposed by regulations and legislations, such as the European Union Council Directive 2008/120/EC5.
Providing meaningful environmental enrichment can be challenging; it should fulfill the natural behavioral needs of the pigs in each stage of their lives, as well as take into consideration practical and technical limitations. Prior to farrowing, providing nesting material is associated with shorter farrowing duration as well as higher survival rate of the newborn piglets during farrowing and throughout the lactation period. Moreover, in free pens with environmental enrichment, maternal behavior is improved, as well as the cognitive performance and weight gain of the piglets3,10,11,12,13. After weaning, grouping pigs (weaners, growing or finishers) from different litters or pens can be stressful and cause aggressiveness towards other pen mates, which may lead to injuries14,15. Therefore, when mixing unfamiliar pigs, providing meaningful environmental enrichment can potentially reduce the occurrence of undesired behavior resulting from frustration and aggressiveness or redirection of rooting behavior.
According to the European Union Council Directive 2008/120/EC (established in 2001/93/EC), it is required that pigs have permanent access to a sufficient quantity of material, to enable proper investigation and manipulation activities, such as straw, hay, wood, sawdust, mushroom compost, peat or a mixture of these5,6. However, the use of these materials may be unmanageable on many farms, as it may cause blockage of the drainage and sewage systems, particularly on farms with slatted flooring. Still, according to the EU Council Directive, farmers can provide alternative enrichment materials, as long as it allows the pigs to perform proper investigation and manipulation activities.
Lack of environmental enrichment may potentially lead to frustration and stress3, which may activate the hypothalamic pituitary adrenal (HPA) axis. In pigs, as well as in humans and other animals, stress typically leads to the secretion of adrenocorticotropic hormone (ACTH) from the pituitary gland. ACTH binds to its receptors on the adrenal cortex and stimulates adrenal release of the glucocorticoid cortisol, which is considered as a major biomarker of stress and can be measured in blood, urine, saliva and hair. Saliva cortisol is a marker for acute stress, since it reflects its biologically active concentration in the blood4. It has the advantage of a non-invasive sample collection, without handling of the animals, by means of cotton ropes provided as environmental enrichment16,17. Hair cortisol is being used as a marker for chronic stress, since cortisol accumulates in the hair over time and can be extracted and measured18,19; however, it requires animal handling, the number of repeated sampling is limited by the hair growth rate, and the analysis is more cumbersome as it require long extraction process. Still, both saliva and hair cortisol may provide complementary information to asses animal welfare.
The objectives of this paper are (1) to give methodologic information on how to provide relatively simple and practical meaningful environmental enrichment for pigs which are housed on slatted floor&#160;during different stages of their lives (farrowing and lactation, weaners to finishers), and (2) to demonstrate how to non-invasively collect saliva samples for the measurement of cortisol concentrations, as a biomarker for acute stress.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Bite-Rite</td>
			<td>Ikadan System USA Inc.</td>
			<td></td>
			<td>Consider ordering additional replaceable silicone sticks</td>
		</tr>
		<tr>
			<td>ELISA; Saliva Cortisol Kit</td>
			<td>DRG International Inc. NJ, USA</td>
			<td>Slv2930</td>
			<td></td>
		</tr>
		<tr>
			<td>HALM 60/80 CM</td>
			<td>W. Domino A/S, DK</td>
			<td>49084/ 85</td>
			<td></td>
		</tr>
		<tr>
			<td>TEGO Swine Oral Fluids Kit</td>
			<td>ITL BioMedical, USA</td>
			<td>A100930</td>
			<td>Including everything needed for the saliva sampling protcol</td>
		</tr>
	</tbody>
</table>

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.

In the current study, 16 litters (170 piglets) were allocated randomly into two treatment groups; in one group, environmental enrichment was provided to eight litters (&#34;Enriched Group&#34;, as described in protocol sections 1-3), while it was not provided to the other eight litters (&#34;Non-enriched Group&#34;). After weaning, each two litters were grouped into one group of about 20 pigs. Saliva samples were collected and analyzed for cortisol concentrations every two weeks in each p...

Watch this Scientific Journal Video about Providing Meaningful Environmental Enrichment and Measuring Saliva Cortisol in Pigs Housed on Slatted Flooring at JoVE.com

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

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 ...

In this video we will demonstrate how to provide practical meaningful environmental enrichment for pigs which are housed on slatted floor during the different stages of their lives and how to collect saliva samples. The environmental enrichment protocols can be implanted easily in almost any swine farm and potentially can improve animal welfare. The saliva sampling is done noninvasively and can be used for cortisol measurements to assess stress or for the measurement of other biomarkers.To begin this procedure hang the enrichment materials in the farrowing pen before the sow is moved into it. For each farrowing pen prepare two pieces of jute and two pieces of 100 percent natural uncolored cotton ropes each approximately one meter long. Tie the jute and cotton ropes to the pen's bars from it's middle allowing the two free ends of each piece to be hung towards the floor.The height of the jute and cotton rope's knots should be just lower than the sow's shoulder while the free ends are just above the floor. After piglets are born hang the jute and cotton ropes so that they are reachable for both the sow and it's piglets in a safe place. Typically, when the materials are reachable for both piglets imitate their dams from a very early age.Shredded paper can also be provided as additional environmental enrichment for the farrowing sows and piglets by spreading it on the slatted floor. For each pen prepare pieces of 100 percent natural uncolored cotton rope that are one meter long making sure to prepare at least one piece for every ten pigs. Tie the cotton ropes to a chain hung from the ceiling or to a hanging hold.Position the ropes at the center of the active area of the pen close to the feeder and drinkers and far from the pen's wall to allow 360 degree access. Hang the ropes at the height which allows the pigs to easily reach it with their mouths. Typically, the knot should be lower than the pig's shoulder while the free ends of the ropes should be just above the floor without touching it.In addition to the cotton ropes provide chewable silicone sticks. The device used contains four chewable sticks which allows at least four pigs to interact with it simultaneously. Hang the chewable device with a chain from the ceiling to allow swinging movement.Position the device at the center of the active area of the pen making sure that the end of the silicone sticks are at a height 10 to 20 centimeters above the floor. First, provide each pen with a straw placed in a rack. Hang the straw racks at a height which is reachable for all pigs.Usually just below the shoulder level and attach them to the wall with the easiest access for as many pigs as possible. Make sure the racks are completely filled with straw at the beginning. A small amount of fresh straw can be added every one to three days.Alternatively, refill the rack when it is empty which typically occurs every seven to ten days. As an alternative to the racks straw can be provided in baskets. Plan saliva sampling for the same time of the day to avoid bias as cortisol secretion changes along the day due to physiologic circadian rhythm or due to feeding.Sampling is typically easier before feeding preferably before the second feeding of the day. In ad libitum feeding plan for the same time of the day. Prepare one piece of uncolored 100 percent cotton rope for each 12 to 15 pigs.Tie the rope close to the center of the pen far from the pen's walls. Make sure that the cord used for hanging is not reachable for chewing by the pigs and that the free ends of the cotton ropes do not touch the floor. Then unravel the free end of the rope to increase the available chewable surface.Leave the rope hanging for 15 to 30 minutes. To extract the saliva from the rope put the free part of the rope into a sterile plastic bag and squeeze it by applying pressure from the outside of the bag from top to bottom until enough liquid accumulates at the bottom of the bag. After this, remove the rope from the bag.Cut the bottom corner of the bag above a 50 milliliter sterile plastic tube and allow the saliva to drain into the tube. Then seal the tube and mark it with either a permanent marker or a pre prepared sticker. Store the saliva samples in an insulated container with ice packs until delivered to the lab.In this study piglets were allocated randomly into two treatment groups. One which received environmental enrichment and one which did not. Saliva cortisol samples were successfully collected every two weeks just before the second feeding of the day.The pigs in the enriched group showed high interest in the saliva collecting cotton ropes from the beginning. However, the non-enriched group saliva volumes are typically lower in the initial collection but seem to be similar between the groups from the second or third collection sessions. Typically, a volume larger than 30 to 40 milliliters is easily extracted from each row.Cortisol concentrations, as measured by Elisa were overall significantly lower in the enriched pigs as compared to the non-enriched pigs. Pigs can be environmentally enriched at all life stages even on slatted floor using jute, cotton ropes, straw in racks and bite-rite devices. The type of enrichment and the way it is located within the pen are important for it's effectiveness.Saliva collected noninvasively by cotton ropes as demonstrated in this video can be used for cortisol measurement as well as other biomarkers or even for the screening of pathogens.

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