Salivary cortisol is a biomarker used to assess stress and physiological responses in horses. It is a glucocorticoid hormone produced by the adrenal cortex and is released into the bloodstream in response to stressors, with measurable levels detectable in saliva. The non-invasive collection of saliva makes it a practical method for evaluating cortisol levels in equine studies. Salivary cortisol is utilized in research to explore stress-related conditions, welfare assessments, and the effects of training or environmental changes on horses. This page compiles peer-reviewed research studies and scholarly articles that investigate the measurement, variability, and implications of salivary cortisol in equine health and behavior.
Reijerkerk EP, Visser EK, van Reenen CG, van der Kolk JH.To compare the effects of IV administration of various doses of ovine corticotrophin-releasing hormone (oCRH) on plasma and saliva cortisol concentrations in healthy horses and determine whether an oCRH challenge test protocol is valid for use in adult horses. Methods: 24 healthy Warmblood horses. PROCEDURES-Each horse received oCRH in saline (0.9% NaCl) via IV administration at a dose of 0 (control treatment), 0.01, 0.1, or 1.0 Mg/kg (6 horses/group). Jugular blood and saliva samples were collected simultaneously 15 minutes before and immediately prior to injection (baseline); data from these...
Brøkner C, Nørgaard P, Hansen HH.The ingestive and post-digestion effect of a blend of special essential oil compounds (EO) on eating, chewing and faecal parameters were measured in horses. Ingestive effects appear after no adaptation. Post-digestion effects appear after adaptation. Six Icelandic horses were assigned to two groups in a Latin Square subplot design with EO treatments to four different roughage types and four different concentrates. The horses were fed four different roughage meals and two different concentrate meals on each of the four sampling days. Eating time and saliva were observed during meals. Jaw moveme...
Carregaro AB, Mataqueiro MI, Soares OA, Queiroz-Neto A.The study of caffeine in racing horses has been of growing concern in veterinary sports medicine since the Association of Racing Commissioners International (ARCI) stated that it has no valid therapeutic use in racehorses. We examined the kinetic alterations in the urinary excretion and salivary secretion of caffeine in seven horses subjected to urinary acidification using ascorbic acid because this procedure can simulate the acidosis that follows anaerobic exercise. They participated in two treatment groups: the control group (SG) received 500 ml of saline and then 2.0 mg kg(-1) caffeine i.v....
Shanahan S.Resistance in the horse to trailer loading is a common source of stress and injury to horses and their handlers. The objective of this study was to determine whether nonaversive training based on the Tellington-Touch Equine Awareness Method (TTEAM; Tellington-Jones &Bruns, 1988) would decrease loading time and reduce stress during loading for horses with a history of reluctance to load. Ten horses described by their owners as "problem loaders" were subjected to pretraining and posttraining assessments of loading. Each assessment involved two 7-min loading attempts during which heart rate and s...
Nicol C.It is frequently asserted that equine stereotypies, such as crib-biting, wind-sucking and weaving, are caused by boredom. However, this explanation is too general to be of practical use in discerning the causes of each stereotypy or in devising management practices to prevent their occurrence. The majority of equine stereotypes start within one month of weaning when both the nutritional and social environment of the foal are substantially altered. Epidemiological research has revealed that the provision of low quantities of forage and minimal opportunities for social contact are associated wit...
Elsaesser F, Klobasa F, Ellendorff F.Previous work (Marc et al., 2000) suggested that plasma cortisol responses to treadmill exercise or ACTH injection are a reliable marker for performance evaluation in warmblood horses. For practical purposes blood sample collections and treadmill exercise tests are somewhat troublesome and time consuming. The goal of this study was thus to evaluate the use of saliva for cortisol determination (by direct EIA) as a marker for performance and to investigate the reliability and repeatability of plasma cortisol responses to a single i.v. injection of ACTH (50 micrograms or 250 micrograms). Furtherm...
Mead DG, Ramberg FB, Maré CJ.In previous experiments we have demonstrated that colonized and wild black flies are competent laboratory vectors of different Mexican and Western USA isolates of vesicular stomatitis virus, serotype New Jersey (VSV-NJ). We have recently demonstrated biological VSV-NJ transmission by black flies using animal models. In the study described here, we tested the vector competence of colonized and wild black flies for the vesicular stomatitis virus, serotype Indiana (VSV-IN). A 1998 equine isolate was used. After a 10 day incubation period, saliva from experimentally infected Simulium vittatum and ...
Sadler VM, Wisner ER, Robertson JT, Moses VS.Congenital anomalies of the equine salivary glands and their ductal systems are rare. In man, parotid duct atresia is thought to be due to a congenital malformation of the first branchial arch. One horse with unilateral parotid salivary duct atresia is described. Imaging modalities available for accurate diagnosis, and treatment options, are reviewed.
Scocco P, Menghi G, Ceccarelli P, Pedini V.This study was aimed at characterizing the glycoconjugates produced by the horse sublingual gland and, in particular, at discriminating between the sialoderivatives by means of differential oxidation and saponification combined with lectin histochemistry and enzymatic degradation. The results showed a predominance of sialoglycoconjugates with beta-galactose as acceptor sugar in the salivary mucins produced by the sublingual gland. Besides being the most represented terminal residue, sialic acid was also expressed in a great variety of derivatives distinguishable on the basis of acceptor sugars...
Newton SA, Knottenbelt DC, Daniel EA.The surgical repair of a traumatic injury to the parotid (Stenson's) duct of an adult horse is described and compared with previous reports. The diagnosis was confirmed by the analysis of a flow of saliva-like fluid observed while the horse was eating. The repair was attempted under general anaesthesia three days after the injury. Surgical anastomosis was considered to have improved the rate of healing. However, there was some evidence of leakage of saliva from the site for three days after the removal of the in-dwelling catheter, seven days postoperatively. A full recovery followed.
Gargiulo AM, Pedini V, Ceccarelli P.The glycoconjugate content of major horse salivary glands was investigated by means of horseradish peroxidase-conjugated lectins. Qualitative differences were observed in the terminal sugar residues of secretory glycoproteins and glycoconjugates linked to the apical surface of excretory duct epithelial cells. Mucous acinar cells in mandibular and sublingual glands contained oligosaccharides with D-galactose, alpha- and beta-N-acetylgalactosamine, N-acetylglucosamine and fucose residues, whereas mandibular, sublingual and parotid serous cells contained only oligosaccharides with terminal alpha-...
Dahlgren LA, DeBowes RM, Gift LJ, Veatch JK.Heterotopic salivary tissue was diagnosed in a 7-month-old Quarter Horse colt with a history of a draining wound in the left temporal region from time of birth. Surgical excision of the tract was performed for diagnosis and treatment. Histologically, tissues were compatible with a mixed-type salivary gland, most likely of parotid salivary gland origin. Complete resolution of the drainage was achieved after surgical removal of the ectopic tissue.
Asari M, Sasaki K, Kano Y, Nishita T.The immunolocalization of carbonic anhydrase isozymes in equine salivary glands was investigated for assessment of their biologic functions. In parotid glands, duct segments showed reactivity with CA-I and CA-III. CA-III was selectively located in duct segments, particularly in the basal cells of the interlobular duct. Serous acinar cells were positive for CA-I and CA-II. In submandibular glands, CA-I and CA-II were present in serous demilune and duct segments. CA-II was selectively located in the duct segments, as also noted in the parotid gland. In sublingual glands, CA-I and CA-II were loca...
Talley MR, Modransky PD, Welker FH, Smith MM, Dubbin ES.A 7-month-old Quarter Horse colt was examined because it had had a tortuous, distended vessel on the side of its head since birth. An abnormality of the parotid salivary duct was suspected on the basis of location and course of the vessel. Atresia of the duct near the parotid papilla was diagnosed by use of contrast sialography. Surgical transpositioning was attempted, but failed because of stricture formation. Chemical ablation of the salivary gland has been used to treat traumatic rupture of the duct. It proved to be an effective and practical method of resolving the problem in this case. Co...
Eckersall PD, Aitchison T, Colquhoun KM.Whole saliva was collected from six horses over a period of five weeks in sufficient volume for the analysis of 10 constituents. There was considerable variation in the concentration of the analytes both between horses and between different days in the same horse. The most variable constituent was sodium, and the least variable was glucose, but this was derived from the sweet used to stimulate salivation. The use of whole saliva as a fluid for investigation would depend on achieving the minimum variability possible.
Hagler WM, Behlow RF.An outbreak of salivary syndrome in horses in North Carolina was investigated. Rhizoctonia leguminicola was the predominant fungus isolated from toxic red clover hay. The fungus was less prevalent in the hay after 10 months of storage, and the hay had also decreased in biological activity after 10 months. Toxic hay caused extreme salivation, piloerection, respiratory distress, and increased frequency of defecation when fed to guinea pigs, and purified extracts of toxic hay and pure slaframine elicited these same responses when injected intraperitoneally into guinea pigs. The freshly acquired h...
Stick JA, Robinson NE, Krehbiel JD.Esophageal fistulas were made in 6 ponies to evaluate whole blood acid-base values and serum and salivary electrolyte alterations associated with salivary depletion. Acid-base and electrolyte values remained within normal ranges for 15 days in 3 control ponies fed a pelleted diet through nasogastric tubes. In 6 ponies with esophageal fistulas that were fed the same diet through esophagostomy tubes, hypochloremia and hyponatremia developed during the same period. Serum K concentrations were only marginally depleted, probably because of dietary replacement. Salivary depletion resulted in transie...
BAER H, KABAT EA, KNAUB V.Blood group substances have been isolated from the saliva of human beings of blood group B and from the linings of individual horse stomachs. The properties of the human B substances are similar to those of hog and human blood group substances previously isolated. The horse substances showed lower hexosamine and reducing sugar and higher total and non-hexosamine nitrogen than do the materials from the other species. Materials isolated from individual horse stomachs possess either A or B activity or both. Certain stomachs yielded products of identical analytical composition but with neither blo...
Hovey MR, Davis A, Chen S, Godwin P, Porr CAS.Stress can impact the health and well-being of animals negatively. Behavioral and physiological changes, particularly serum cortisol, offer objective and easy-to-use methods of evaluating stress in horses. However, limited studies support a positive relationship between changes in stress-related serum cortisol concentrations and stress-related behaviors in horses. This study assessed differences in stress-related behaviors and serum cortisol concentrations in horses used in a therapeutic riding program (TRH) or university riding program (UNI). Riders were grouped by disability type (TRH) or by...
Sadler VM, Wisner ER, Robertson JT, Moses VS.Congenital anomalies of the equine salivary glands and their ductal systems are rare. In man, parotid duct atresia is thought to be due to a congenital malformation of the first branchial arch. One horse with unilateral parotid salivary duct atresia is described. Imaging modalities available for accurate diagnosis, and treatment options, are reviewed.
Talley MR, Modransky PD, Welker FH, Smith MM, Dubbin ES.A 7-month-old Quarter Horse colt was examined because it had had a tortuous, distended vessel on the side of its head since birth. An abnormality of the parotid salivary duct was suspected on the basis of location and course of the vessel. Atresia of the duct near the parotid papilla was diagnosed by use of contrast sialography. Surgical transpositioning was attempted, but failed because of stricture formation. Chemical ablation of the salivary gland has been used to treat traumatic rupture of the duct. It proved to be an effective and practical method of resolving the problem in this case. Co...
Newton SA, Knottenbelt DC, Daniel EA.The surgical repair of a traumatic injury to the parotid (Stenson's) duct of an adult horse is described and compared with previous reports. The diagnosis was confirmed by the analysis of a flow of saliva-like fluid observed while the horse was eating. The repair was attempted under general anaesthesia three days after the injury. Surgical anastomosis was considered to have improved the rate of healing. However, there was some evidence of leakage of saliva from the site for three days after the removal of the in-dwelling catheter, seven days postoperatively. A full recovery followed.
Campbell MLH, Hampshire D, Hamstead LE, Rose BV, Smith KC, de Mestre AM.Intrauterine infusion of peanut oil at Day 10 post-ovulation has been reported to prolong dioestrus in mares. However, the effects of peanut oil treatment on the endometrium and whether the technique is painful have not been assessed. The objectives of this study were, (i) to determine the effect of intrauterine infusion of peanut oil on endometrial health, (ii) to determine whether use of intrauterine peanut oil is painful and (iii) to confirm that peanut oil causes prolonged dioestrus. Six mares aged 3-12 years old were used in a cross-over design with each mare administered both 1 ml of in...
Bannai H, Kambayashi Y, Ohta M, Nemoto M, Tsujimura K.To assess the prevalence of equine coronavirus infection in riding horses, virus-neutralizing tests were performed on serum and saliva samples collected at four facilities in Japan. Seropositivity rates ranged from 79.2% to 94.6%, suggesting widespread circulation of the virus in these populations. Antibody prevalence in saliva samples from two facilities that had experienced outbreaks in the previous year (67.6% and 71.4%) was significantly higher than at the other facilities without reported outbreaks (41.7% and 45.2%, P<0.05). The presence of salivary antibodies in a high proportion of h...
Melchert M, Aurich C, Aurich J, Gautier C, Nagel C.Stress at foaling has been demonstrated to delay birth. In this study, we followed the hypothesis that even a short delay of foaling increases catecholamine and cortisol release in foals, induces acidosis and impairs neonatal adaptation. Foaling was prolonged for 5 min by transferring mares to an unfamiliar environment at rupture of the allantochorion (group delay, n = 6) while control mares (n = 5) were left undisturbed. In their foals, times from birth to first standing and first suckling, heart rate, heart rate variability (HRV) and salivary cortisol concentration were analysed. B...
Clothier J, Small A, Hinch G, Brown WY.The persistent endocrinological effects of perinatal stress due to gestational immaturity in horses are unknown, although effects have been reported in other livestock species. This pilot study tested the hypothesis that persistent adrenocortical dysregulation is present in horses that were gestationally immature at birth by assessing the salivary cortisol response to exogenous ACTH. Case horses (n = 10) were recruited with histories of gestation length < 315 d or dysmaturity observable through neonatal signs. Positive controls (n = 7) and negative controls (n = 5) were recruited wher...
Ekici S, Ozmen O.The active roles of microribonucleic acids (miRNAs) in gene regulation have made miRNAs a key point for the scientific world in the study of physiological processes. Although saliva includes the largest number of miRNAs, there is no miRNA study in saliva on horses has been found. Our study is the first study on miRNAs isolation from saliva in horses. In the present study, saliva was studied in Arabian racehorses to better understand the molecular mechanisms of expression levels that are effective in lipid metabolism of miRNAs and their target genes during the race. Identification of lipid meta...
Kimura R, Borankulova S, Maratbek SZ.This study aimed to evaluate the effect of differences in trainer skills on horse training during the early stages of riding habituation by measuring the levels of stress and changes in stress levels. Among nine untrained horses employed, five in Group A were trained by two low-skilled trainers, whereas the remaining four in Group B were trained by two high-skilled trainers using the traditional Kazakh method. Salivary α-amylase concentration was measured as a biomarker of stress immediately before and after each riding session during the training period. In the duration of riding and mooring...
Becker-Birck M, Schmidt A, Wulf M, Aurich J, von der Wense A, Möstl E, Berz R, Aurich C.Bringing the head and neck of ridden horses into a position of hyperflexion is widely used in equestrian sports. In our study, the hypothesis was tested that hyperflexion is an acute stressor for horses. Salivary cortisol concentrations, heart rate, heart rate variability (HRV) and superficial body temperature were determined in horses (n = 16) lunged on two subsequent days. The head and neck of the horse was fixed with side reins in a position allowing forward extension on day A and fixed in hyperflexion on day B. The order of treatments alternated between horses. In response to lunging, ...
Massányi M, Halo M, Mlyneková E, Kováčiková E, Tokárová K, Greń A, Massányi P, Halo M.The performance of sport horses is conditioned not only by the quality of its gene pool, but also by a large number of external factors. The most dominant being nutrition, quality of breeding, level of zootechnical care and the quality of the sports rider and coach. Important factor is the process of individuals' adaptation to the training load occurring during the training itself. This study was focused on the analysis of salivary cortisol levels as well as hematological and biochemical blood parameters in relation to load to which the tested horses were subjected. In the study 14 horses of s...
Hensel B, Jakop U, Schmicke M, Schröter F, Jung M, Schulze M.Artificial insemination (AI) is commonly used in the equine industry to enhance the genetic value in breeding programs and to effectively utilize ejaculates. Many stallions are used as breeding stallions as well as in high-level sports competitions to improve their market value. The goal of the present study was to investigate whether this dual use of stallions influences the animals´ stress levels and/or the quality of their ejaculates. For this purpose, 18 stallions were grouped into two categories: breeding stallions with (BSC = breeding stallion competition), and breeding stallions with...
Schönbom H, Kassens A, Hopster-Iversen C, Klewitz J, Piechotta M, Martinsson G, Kißler A, Burger D, Sieme H.Pregnancy diagnostics in equine reproduction are routinely performed using transrectal ultrasonography, although it is also possible to visualize the fetus by transabdominal ultrasound examinations from the 90th day of gestation onward. We hypothesized that ultrasound examinations may stress the mare and that the gestational stage status and lactation may influence the mare's stress reaction. To investigate the stress reaction, 25 thoroughbred mares of different age, pregnancy and lactational status underwent a transrectal examination. In pregnant mares, an additional transabdominal examinatio...
Mead DG, Ramberg FB, Maré CJ.In previous experiments we have demonstrated that colonized and wild black flies are competent laboratory vectors of different Mexican and Western USA isolates of vesicular stomatitis virus, serotype New Jersey (VSV-NJ). We have recently demonstrated biological VSV-NJ transmission by black flies using animal models. In the study described here, we tested the vector competence of colonized and wild black flies for the vesicular stomatitis virus, serotype Indiana (VSV-IN). A 1998 equine isolate was used. After a 10 day incubation period, saliva from experimentally infected Simulium vittatum and ...
Keating DL, Lehman JS, Burk SV.Identification of factors associated with parasite infestation in horses could reduce frequency of anthelmintic administration and slow development of anthelmintic resistance. The study aim was to evaluate management factors, equine characteristics, and equine salivary cortisol concentrations for association with strongyle-type egg shedding levels. As immunocompromised horses appear to be more susceptible to parasite infestation, it was hypothesized that salivary cortisol concentration could be associated with parasite egg shedding. Saliva and fecal samples were collected from 200 horses acros...
Jacquay ET, Harris PA, Adams AA.Transportation of horses on short journeys can lead to an increase in stress. There are known age-associated changes in immune and metabolic responses in horses; however, no research exists evaluating how age may influence these responses to transportation stress. Eleven mares within two age groups, aged (n = 5, 22 ± 1 year) or young (n = 6, 2 ± 1 year), were transported 1 hour and 20 minutes. Peripheral blood and saliva were collected before and after transportation at baseline (2 to 3 weeks prior to transportation), 24 hours pre-transport, 1 hour before loading, 15 minutes, 30 minutes,...
Elsaesser F, Klobasa F, Ellendorff F.Previous work (Marc et al., 2000) suggested that plasma cortisol responses to treadmill exercise or ACTH injection are a reliable marker for performance evaluation in warmblood horses. For practical purposes blood sample collections and treadmill exercise tests are somewhat troublesome and time consuming. The goal of this study was thus to evaluate the use of saliva for cortisol determination (by direct EIA) as a marker for performance and to investigate the reliability and repeatability of plasma cortisol responses to a single i.v. injection of ACTH (50 micrograms or 250 micrograms). Furtherm...
Muñoz-Prieto A, Cerón JJ, Rubio CP, Contreras-Aguilar MD, Pardo-Marín L, Ayala-de la Peña I, Martín-Cuervo M, Holm Henriksen IM....In this report, the measurement of salivary biomarkers as an aid for diagnosis of equine gastric ulcer syndrome (EGUS) was studied. A comprehensive panel of 23 salivary analytes was measured in the saliva of horses affected by EGUS and compared to healthy animals and horses with other diseases clinically similar to EGUS but with a negative diagnosis at gastroscopic examination. A total of 147 horses were included in the study and divided into heathy population (n = 12), the EGUS group (n = 110), and the group of horses with other diseases (n = 25). From the 23 analytes studied, 17 showed incre...
Scheidegger MD, Gerber V, Bruckmaier RM, van der Kolk JH, Burger D, Ramseyer A.This study tested the hypothesis that adrenocortical function would be altered in horses with equine gastric ulcer syndrome (EGUS). Twenty-six sport horses competing at national or international levels in eventing (n=15) or endurance (n=11) were subjected to a gastroscopic examination and an adrenocorticotropic hormone (ACTH) stimulation test. Salivary cortisol concentrations were measured before (baseline) and after (30, 60, 90, 120 and 150min) IV ACTH injection (1μg/kg bodyweight). Within EGUS, two distinct diseases, equine squamous gastric disease (ESGD) and equine glandular gastric diseas...
Brøkner C, Nørgaard P, Hansen HH.The ingestive and post-digestion effect of a blend of special essential oil compounds (EO) on eating, chewing and faecal parameters were measured in horses. Ingestive effects appear after no adaptation. Post-digestion effects appear after adaptation. Six Icelandic horses were assigned to two groups in a Latin Square subplot design with EO treatments to four different roughage types and four different concentrates. The horses were fed four different roughage meals and two different concentrate meals on each of the four sampling days. Eating time and saliva were observed during meals. Jaw moveme...
Wood MK, Mason RM, Burk SV, Birmingham SSW.The goal of this study was to determine whether oral administration of α-casozepine (Zylkene) supplementation decreases the risk of disease in foals, as measured through salivary cortisol levels, temperature, clinical signs of disease, and body weight before and after weaning. Methods: 20 Thoroughbred foals were observed over 10 weeks, beginning 14 days prior to the time of weaning. Methods: Experimental foals were randomly selected and given α-casozepine daily for 10 weeks, while the controls received a placebo with the same frequency as the experimental group. The average daily height and ...
Villalba-Orero M, Contreras-Aguilar MD, Cerón JJ, Fuentes-Romero B, Valero-González M, Martín-Cuervo M.Stress leukogram includes eosinopenia as one of its main markers (neutrophilia, eosinopenia, lymphopenia, and mild monocytosis). Cortisol is the main stress biomarker, which is also strongly correlated with the severity of gastrointestinal diseases. This study aimed to determine the relationship between salivary cortisol and the eosinophil cell count (EC) in equids with abdominal pain. To do this, 39 horses with abdominal pain referred to an emergency service were included. All samples were taken on admission, and several parameters and clinical data were included. Equids were classified accor...
