Simultaneous detection and quantification of six equine cytokines in plasma using a fluorescent microsphere immunoassay (FMIA)

Cytokines are cell signalling proteins that mediate a number of different physiological responses. They are also biomarkers for inflammatory conditions and potential diagnostic references for diseases. Until recently, simultaneous quantification of cytokine profiles had not been possible. Now however, fluorescent microsphere immunoassays (FMIA) are able to measure multiple cytokines in a single sample. The following pro-inflammatory and anti-inflammatory cytokines were quantified in equine plasma and serum samples: interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α. • The objective of this study was to quantify six equine cytokines simultaneously using the BioPlex® 200 system in equine EDTA-plasma and serum. • It demonstrates an increased number of detectable cytokines over published studies. • This technology has the advantage of reduced sample volume and assay time compared to traditional sandwich ELISAs

Using the Horse Grimace Scale (HGS) to Assess Pain Associated with Acute Laminitis in Horses (Equus caballus)

Acute laminitis is a common equine disease characterized by intense foot pain, both acutely and chronically. The Obel grading system is the most widely accepted method for describing the severity of laminitis by equine practitioners, however this method requires movement (walk and trot) of the horse, causing further intense pain. The recently developed Horse Grimace Scale (HGS), a facial-expression-based pain coding system, may offer a more effective means of assessing the pain associated with acute laminitis. The aims of this study were: to investigate whether HGS can be usefully applied to assess pain associated with acute laminitis in horses at rest, and to examine if scoring HGS using videos produced similar results as those obtained from still images. Ten horses, referred as acute laminitis cases with no prior treatment, were included in the study. Each horse was assessed using the Obel and HGS (from images and videos) scales: at the admission (before any treatment) and at seven days after the initial evaluation and treatment. The results of this study suggest that HGS is a potentially effective method to assess pain associated with acute laminitis in horses at rest, as horses showing high HGS scores also exhibited higher Obel scores and veterinarians classified them in a more severe painful state. Furthermore, the inter-observer reliability of the HGS total score was good for both still images and video evaluation. There was no significant difference in HGS total scores between the still images and videos, suggesting that there is a possibility of applying the HGS in clinical practice, by observing the horse for a short time. However, further validation studies are needed prior to applying the HGS in a clinical setting

Circulating β-endorphin, adrenocorticotrophic hormone and cortisol levels of stallions before and after short road transport: stress effect of different distances

<p>Abstract</p> <p>Background</p> <p>Since transport evokes physiological adjustments that include endocrine responses, the objective of this study was to examine the responses of circulating β-endorphin, adrenocorticotrophic hormone (ACTH) and cortisol levels to transport stress in stallions.</p> <p>Methods</p> <p>Forty-two healthy Thoroughbred and crossbred stallions were studied before and after road transport over distances of 100, 200 and 300 km. Blood samples were collected from the jugular vein: first in a single box immediately before loading (pre-samples), then immediately after transport and unloading on arrival at the breeding stations (post-samples).</p> <p>Results</p> <p>An increase in circulating β-endorphin levels after transport of 100 km (<it>P </it>< 0.01), compared to basal values was observed. Circulating ACTH levels showed significant increases after transport of 100 km (<it>P </it>< 0.001) and 200 km (<it>P </it>< 0.001). Circulating cortisol levels showed significant increases after road transport over distances of 100, 200 and 300 km (<it>P </it>< 0.001). An effect of transport on β-endorphin, ACTH and cortisol variations was therefore evident for the different distances studied. No significant differences (<it>P </it>> 0.05) between horses of different ages and different breeds were observed for β-endorphin, ACTH and cortisol levels.</p> <p>Conclusion</p> <p>The results obtained for short term transportation of stallions showed a very strong reaction of the adrenocortical system. The lack of response of β-endorphin after transport of 200–300 km and of ACTH after transport of 300 km seems to suggest a soothing effect of negative feedback of ACTH and cortisol levels.</p

Awin welfare assessment protocols

What are the AWIN welfare assessment protocol? It is a science-based welfare assessment method, built on animal-based indicators, including pain. What are they for? It is intended to assess welfare of turkeys, sheep, goats, horses and donkeys on farm. Why use a two level approach strategy? The AWIN welfare assessment protocols offer, as a first level, a quick screening, consisting of a selection of robust and feasible animal-based indicators. Depending on the outcome, a second level, consisting of a more comprehensive and in-depth assessment, may be recommended

Development of a facial expressions of pain scale in horses undergoing routine castration

The objective of this study was to develop and validate a standardized pain scale based on facial expressions of horses (Horse Grimace Scale HGS). Forty stallions of different breeds aged 1\uf75 years underwent routine surgical castration with closed technique under general anaesthesia. They were assigned to one of two treatment groups balanced for breed. A (n= 19) received one single injection of Flunixin-Meglumine immediately before anesthesia. B (n= 21) received Flunixin-Meglumine before anesthesia and six hours after the surgery. As a control group (C) horses (N=6) needing noninvasive indolent diagnostic procedures under general anesthesia were recruited. None of the subjects had had pre-existing painful conditions. All horses were hospitalized in an clinic for 5 days in an observation box with two HD video-cameras positioned on the top opposite sides of the box. Thirty-minute videos were recorded on the day prior to surgery (baseline) and at 8 hours after anesthesia. From each video high quality images of the horses\u2019 head were extracted. These images were compared to identify changes in facial expressions (see Langford et al, 2010) by a treatment-blind observer experienced in assessing facial expressions in other species. Six facial action units were defined (stiffly backwards ears, orbital tightening, tension above the eye area, strained chewing muscles, mouth strained and pronounced chin, strained nostrils and flattening of the profile). 60 pre and 60 post treatment images (randomly selected) were then scored in a random sequence using the action units by five treatment-blind participants. The mean HGS was analysed using a GLM for repeated measures with time (pre-vs-post treatment) as the within-subjects factor and treatment as the between subjects factor. 73,3% of images were correctly assessed as true post-surgery or true basal. Inter-class correlation coefficient among observers was 0,91 showing high inter-observer reliability. A significant effect of time, treatment and time*treatment were found (P<0,001). A and B significantly differed from C (P<0,01). LP: A new approach to assess pain based on a standardized scale of facial expressions was developed. It has the advantages of being not time consuming, easily trainable and applicable

Measuring validity and reliability of facial expressions of pain in horses as an innovative welfare indicator

This study aimed to develop and validate a standardized pain scale based on facial expressions of horses (Horse Grimace Pain Scale, HGPS). Forty stallions of different breeds aged 1\uf75 years were assigned to one of two groups. A (n= 19) underwent routine surgery castration in general anaesthesia and received one injection of Flunixin-Meglumine immediately before anaesthesia. B (n= 21) underwent the same surgical procedure but received Flunixin-Meglumine before anesthesia and six hours after the surgery. Six horses with no pain but needing noninvasive, indolent diagnostic procedures received the same treatment than A but not surgery and were used as controls (C). The horses were hospitalized at the Havelland Clinic (see abstract \u201cPain assessment in horses after routine castration\u201d) for 5 days. Two digital video-cameras were positioned on the top opposite sides of the box. Thirty-minutes videos were recorded on the day prior to surgery (baseline) and at 8 hours following castration. From each video images of the horses\u2019 faces were extracted, enabling generation of a number of clear and high quality pictures. Images were compared to identify changes in facial expressions associated with castration pain by a treatment-blind observer experienced in assessing facial expressions in other species. Six facial action units were defined. Baseline and 8-hours post procedure images of each horse were selected at random. 126 images were then scored in a randomized sequence using the action units by five treatment-blind participants. In order to explore the effect of time and treatment, the horse grimace pain scale (HGPS) for each image was determined by summarizing the score provided for each action unit. 73,3% images were correctly assessed as true post-surgery or true basal. Inter-class correlation coefficient among observers was 0,91 showing high inter-observer reliability. GLM for repeated measures showed a significant effect of time and treatment (P<0,001), A and B significantly differed from C (P<0,01). A promising welfare indicator based on a standardized scale of facial expressions showed validity and reliability. It has the advantages of being not time consuming, easily trainable and applicable on farm