Effects of a supplement containing hyaluronan with TBG‐136™ schizophyllan β‐glucan on gastric ulcer scores in horses

BackgroundEquine gastric ulcer syndrome (EGUS) is a frequent disease in horses that comprises two different entities: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). This disease considerably reduces the quality of life of affected horses and can negatively affect performance. Saliva contains biomarkers, such as oxytocin, that have been used as a welfare indicator and can develop a function as a protective factor against stress‐induced changes in gastric function due to its gastric antisecretory and antiulcer effects. The objective of this work was to evaluate changes in salivary oxytocin concentrations in healthy and EGUS horses. For this purpose, an immunoassay based on AlphaLISA technology was validated for the quantification of salivary oxytocin and applied in a total of 102 horses divided into 5 groups: 25 with both EGUS, 23 with only EGGD, 21 with only ESGD, 19 horses with other diseases, and 14 healthy horses.ResultsThe analytical validation of the method showed good precision and linearity under dilution. Salivary oxytocin concentrations in healthy horses were higher compared to horses with both ESGD and EGGD and only EGGD. Salivary oxytocin concentrations in horses with only ESGD were higher compared to horses with both ESGD and EGGD and horses with only EGGD. In addition, salivary oxytocin concentrations in horses with other diseases different from ESGD were significantly increased compared to horses with both ESGD and EGGD and horses with only EGGD.ConclusionsThis report validates a new assay that can measure oxytocin in saliva in horses in a precise and accurate way. The lower oxytocin values in horses with EGGD and both EGGD and ESGD than in horses with ESGD, horses with other diseases, and healthy horses could indicate a possible relation of oxytocin with this disease.

BackgroundTo date, risk factors for equine glandular gastric disease (EGGD) have not been described in Thoroughbred racehorses.ObjectivesTo determine management factors associated with EGGD, identify clinical signs in affected horses, and compare these to equine squamous gastric disease (ESGD).AnimalsThe study was carried out on 109 Thoroughbred racehorses from 8 training yards (3 in the United Kingdom and 5 in Australia).MethodsGastroscopic examination alongside a questionnaire regarding management, feeding, exercise, and health.ResultsManagement factors and clinical signs were different for EGGD versus ESGD. Exercising ≥5 days per week was associated with a 10.4 times (95% CI [confidence interval]: 1.34‐26.9) increased risk of EGGD. Horses racing below expectation were 3.7 times (95% CI: 1.1‐16.7) more likely to have EGGD. Trainer was also identified as a risk factor for EGGD. Time in work ≤6 weeks was associated with a decreased risk of ESGD (odds ratio [OR] 0.3; 95% CI: 0.1‐0.99). Horses aggressive to humans were less likely to have ESGD (OR 0.12; 95% CI: 0.03‐0.54). Horses with stereotypies were more likely to have ESGD (OR 5.0; 95% CI: 1.6‐15.9).Conclusions and Clinical ImportanceThe findings of our study further support the notion that EGGD should be considered as a distinct disease entity to ESGD. Exercising ≤4 days per week could reduce the risk of EGGD. Horses with EGGD are more likely to perform below expectation and, as such, EGGD might be performance limiting in some affected individuals. Stress minimization could reduce the risk of EGGD.

Simple SummaryEquine gastric ulcer syndrome (EGUS) is a frequent disease that considerably reduces the quality of life of horses, while also affecting their physical performance, even when the disease is subclinical. In this study, we hypothesized that two analytes, calprotectin (CALP) and aldolase, could be measured in saliva with commercially available assays and be potential biomarkers of EGUS in horses. We investigated the changes in the salivary CALP and aldolase in 131 horses divided into 5 different groups: controls (healthy horses), with equine squamous gastric disease (ESGD), with equine glandular gastric disease (EGGD), with combined ESGD and EGGD, and horses with other intestinal pathologies but clinical signs similar to EGUS. The two assays were precise and accurate, and, in both cases, they showed differences between horses with EGUS and healthy horses, but they did not show significant differences between horses with EGUS and horses with other diseases. Our data showed that CALP has a high ability to differentiate between healthy horses and horses with EGUS. Therefore, it could have potential use as a biomarker since a value of CALP in the range of healthy individuals could indicate that the animal is not likely to have EGUS at gastroscopy.Equine gastric ulcer syndrome (EGUS) is a highly prevalent disease that affects horses worldwide. Within EGUS, two different forms have been described: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). The associated clinical signs cause detrimental activity performance, reducing the quality of life of animals. Saliva can contain biomarkers for EGUS that could be potentially used as a complementary tool for diagnosis. The objective of this work was to evaluate the measurements of calprotectin (CALP) and aldolase in the saliva of horses as potential biomarkers of EGUS. For this purpose, automated assays for the quantification of these two proteins were analytically validated and applied for detecting EGUS in a total of 131 horses divided into 5 groups: healthy horses, ESGD, EGGD, combined ESGD and EGGD, and horses with other intestinal pathologies. The assays showed good precision and accuracy in analytical validation, and they were able to discriminate between horses with EGUS and healthy horses, especially in the case of CALP, although they did not show significant differences between horses with EGUS and horses with other diseases. In conclusion, salivary CALP and aldolase can be determined in the saliva of horses and further studies are warranted to elucidate the potential of these analytes as biomarkers in EGUS.

The aims of this study were to better characterize the adrenal response to i.v. adrenocorticotropic hormone (ACTH) in horses with and without gastric disease and to validate and simplify the ACTH stimulation test by determining the diagnostic accuracy of six consecutive sampling time points after ACTH administration for equine glandular gastric disease (EGGD) and equine squamous gastric disease (ESGD). Twenty-six endurance and eventing horses without clinical disease [Sport Horse Population (SHP)] and an independent population of 62 horses [General Population (GP)] were grouped by gastroscopic findings (no/mild vs. moderate/severe EGGD, grade 0-1 vs. 2-4 ESGD, respectively) and underwent an ACTH stimulation test. Salivary cortisol (ng/mL) was analyzed before and 30, 60, 90, 120, and 150 min after i.v. injection of 1 µg/kg BW synthetic ACTH1-24. The association between having moderate or severe EGGD or ESGD and the amount of salivary cortisol was analyzed by means of receiver-operating characteristic (ROC) analysis. The following explanatory variables were considered: cortisol values for every time point, the area under the curve (AUC)-including all time points and corrected for the baseline-and the partial areas under the curve AUC0-90 and AUC90-150. Sampling after 60 min had highest association with moderate/severe EGGD. The diagnostic potential of the ACTH test was higher for the SHP [sensitivity 100% (95% CI 54% to 100%), specificity 75% (95% CI 51% to 91%), ROC-AUC 91% (95% CI 69% to 98%), 1-sided P-value < 0.001] than for the GP [sensitivity 75% (95% CI 48% to 93%), specificity 52% (95% CI 37% to 67%), ROC-AUC 68% (95% CI 51% to 79%), 1-sided P-value = 0.0064]. There were, however, no significant associations with ESGD. The superiority of sampling after 60 min suggests that the initial release of cortisol rather than its peak or the AUC are relevant regarding EGGD. Even though the wide confidence intervals and thus the lack of diagnostic accuracy do not presently support clinical use, characterization of the adrenal response to an ACTH stimulus improves the understanding of EGGD pathophysiology and its relation to stress.

The objective of this study was to evaluate the possible use of spectrophotometric assays for the measurement of trace elements, including Zinc (Zn), Copper (Cu), Magnesium (Mg), and iron (Fe) in the saliva of horses and study their possible changes in equine gastric ulcer syndrome (EGUS). EGUS is a highly prevalent disease, with a current high incidence due to the increase in intensive management conditions. There are two EGUS diseases: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD), which can appear individually or together. For this purpose, automated spectrophotometric assays for measuring these analytes in horse saliva were analytically validated. Then, these analytes were measured in the saliva of horses with only ESGD, only EGGD, both ESGD and EGGD and a group of healthy horses. The methods used to measure the analytes were precise and accurate. Horses diagnosed with EGGD presented significantly lower levels of Zn and Mg. Fe concentrations were significantly lower in the saliva of horses with ESGD and EGGD. Overall, these results indicate that there are changes in trace elements in saliva in EGUS that could reflect the physiopathological mechanisms involved in this process and open the possibility of using trace elements as biomarkers of this syndrome.

Oral omeprazole is the accepted treatment for equine squamous gastric disease (ESGD); however, it is not universally effective. Esomeprazole results in more consistent and pronounced acid suppression in men and is more effective than omeprazole in the treatment of oesophageal and gastric disease. Pharmacodynamic and pilot clinical studies have indicated esomeprazole might also be more effective than omeprazole in horses. To compare the efficacy and safety of oral esomeprazole and omeprazole pastes in the treatment of ESGD and, where present, concurrent equine glandular gastric disease (EGGD). Randomised, single-blinded controlled trial. Horses presenting with grade ≥2 ESGD lesions were randomly allocated to receive 4 mg/kg of either a buffered esomeprazole or omeprazole paste orally once daily for 28 days before gastroscopy being repeated within a further 3 days. Videos and images were anonymised and subsequently graded blind by one researcher. The severity of ESGD (and EGGD) lesions before and after treatment, and thereby treatment responses, were compared using univariable logistic regression. A higher proportion of horses had ESGD healing in response to esomeprazole treatment (63/74, 85%) than with omeprazole treatment (43/73, 59%) (odds ratio [OR]: 4.00, 95% confidence interval [CI]: 1.81, 8.82, p = 0.001). In a subset of horses that had concurrent EGGD, a greater proportion of the horses treated with esomeprazole had lesions ≤grade 1 (esomeprazole 28/51, 55%; omeprazole 6/24, 25%; OR: 3.65, 95% CI: 1.25, 10.71, p = 0.02) Using grade 0 as the benchmark for EGGD healing, the difference remained significant (OR: 4.44, 95% CI: 1.33, 14.85, p = 0.02). It may not be possible to extrapolate these results to other populations with different signalment or management. Oral-buffered esomeprazole was a more effective treatment for ESGD (and concurrent EGGD) than oral-buffered omeprazole.

Prevalence of, and risk factors for, equine squamous gastric disease (ESGD) are well established. Limited data exists on risk factors for equine glandular gastric disease (EGGD). To identify management factors associated with EGGD in show jumping Warmbloods in training. A secondary objective was to identify management factors associated with ESGD. Cross-sectional. Gastroscopies were performed in horses following a 12-16 h fast. Management questionnaires were collected for each horse. Risk factors were determined using multivariable logistic regression modelling. Eighty-three horses were included in the final analysis. Exercising ≥6 days per week increased the odds of EGGD grade ≥1/4 (odds ratio [OR] = 3.5; 95% confidence interval [CI] 1.2-10.7) compared to less frequent exercise. Currently showing increased the risk of EGGD grade ≥2/4 (OR = 10.2; 95% CI, 1.04-100), while competing at the international level decreased the odds of EGGD grade ≥2/4 (OR = 0.11; 95% CI, 0.01-0.97). Exercise intensity increased the odds of grade ≥1/4 ESGD (OR = 2.8; 95% CI, 1.03-7.8) and feeding beet pulp decreased odds (OR = 0.22; 95% CI, 0.07-0.7). Exercise intensity (OR = 3.8; 95% CI, 1.1-12.8) increased the likelihood of grade ≥2/4 ESGD and feeding beet pulp decreased the odds of grade ≥2/4 ESGD (OR = 0.1; 0.02-0.64) respectively. This study used a convenience sample of horses within a relatively small (approximately 200 km) geographic radius. The sample size was relatively small, particularly within the international competition level group. Training and feeding strategies and competition level appear to influence the occurrence of EGGD and ESGD. Prospective studies evaluating the impact of training frequency, duration, and intensity on gastric physiology may clarify the role of exercise in gastric disease.

To investigate equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD) in Icelandic horses moving from pasture into training. 81 horses (median age, 3 years; interquartile range, 1 year) from 10 farms representing 4 different Icelandic regions. Initial gastroscopy was undertaken within 2 weeks of moving from pasture into a training establishment. A total of 71 horses underwent endoscopic examination again 8 weeks later. Various management and behavioral factors were assessed through face-to-face questionnaires with the owners or trainers. Multivariable logistic regression was used to determine factors contributing to any change in ESGD and EGGD severity score during the 8-week training period. Incidence of EGGD and ESGD in this feral population was similar to that found in domesticated horses. ESGD incidence (severity score, ≥ 2; score range, 0 to 4) reduced from an initial 71.6% (58/81) to 25.4% (18/71). On multivariable analysis, sex (ie, being a stallion or a female vs gelding) increased the likelihood of ulcer grade reduction. Being fed preserved forage 3 or more times a day also improved the likelihood of ESGD reduction (odds ratio, 17.95; 95% CI, 1.67 to 193.40; P = .017). Overall, the farm explained 35% of the variance, confirming the importance of management factors. Incidence of EGGD (severity score, ≥ 1; score range, 0 to 2) reduced from 47% (38/81) to 40.8% (29/71) during the same period. No measured variables were associated significantly with EGGD incidence or reduction. Pasture provision (without supplementary feed or forage) does not result automatically in a low incidence of gastric ulcers. Regular provision of preserved forage is a key factor in reducing ESGD incidence.

To investigate the role of equine herpesvirus-2 (EHV-2) and equine herpesvirus-5 (EHV-5) in equine glandular gastric disease (EGGD) by visualizing and quantifying these gamma herpesviruses in EGGD-affected and normal glandular gastric mucosa of horses. A secondary objective was to describe the histopathological abnormalities in the equine gastric glandular mucosa in horses with EGGD. 29 horses (n = 21 postmortem and 8 gastroscopy) categorized as normal (11), EGGD (12), or both EGGD and equine squamous gastric disease (6). Glandular gastric mucosal samples were collected from horses by gastroscopy or postmortem. Histopathology and in situ hybridization targeting EHV-2 and EHV-5 were performed on grossly normal and abnormal glandular gastric mucosa. The number of in situ hybridization-positive cells per millimeter squared of tissue was calculated. Evaluators were blinded to groups. Glandular gastric tissues from horses without EGGD had higher viral loads in the mucosa than normal or abnormal tissues from EGGD horses. There was no difference in viral loads for EHV-2 or EHV-5 between grossly or endoscopically normal to abnormal gastric tissues within horses with EGGD. Lymphocytic plasmacytic gastritis was the most common histopathological abnormality, with only 3 horses having mucosal disruption (glandular ulcer or erosion). Equine gamma herpesviruses are unlikely to play a role in the pathophysiology of EGGD. EGGD is frequently inflammatory with occasional mucosal disruption (ulcer or erosion).

An extended-release injectable omeprazole formulation (ERIO) has become a popular treatment for equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD) where it is available; however, published data are limited and optimal treatment regimens have not been determined. To compare effects of treatment on ESGD and EGGD when an ERIO formulation is administered at either 5- or 7-day intervals. Retrospective clinical study. Case records and gastroscopy images of horses with ESGD or EGGD treated with ERIO were reviewed. Images were anonymised and graded by one researcher masked to treatment group. Treatment responses were compared between the two treatment schedules using univariable ordered logistic regression. Forty-three horses were treated with ERIO at 5-day intervals and 39 horses at 7-day intervals. Signalment and presenting signs did not differ between groups. The proportions of horses with EGGD healing (to grade 0 or 1) in association with ERIO used at 5-day intervals (93%) were higher than associated with treatment at 7-day intervals (69%; odds ratio [OR]: 2.41, 95% CI: 1.23-4.74, p = 0.01). For ESGD, there was no significant difference in the proportion of horses healing in association with treatment at 5-day intervals (97%) compared with 7-day intervals (82%; OR: 2.75, 95% CI: 0.91-8.31, p = 0.07). Four of 328 injections were associated with an injection-site reaction (1%). Retrospective study design, lack of randomisation and limited case numbers. The use of ERIO at 5-day intervals might be more appropriate than the 7-day interval that is used currently.

IntroductionEquine Gastric Ulcer Syndrome (EGUS) is a highly prevalent disorder in horses, which can be classified, based on the localization of the lesions, as Equine Squamous Gastric Disease (ESGD) or Equine Glandular Gastric Disease (EGGD). Although EGUS is recognized as a common cause of poor performance in racehorses, objective investigations about its relation with athletic capacity are lacking. Therefore, the present retrospective study aims to evaluate the associations between EGUS severity and some fitness parameters measured during an incremental treadmill test in Standardbred racehorses in training.MethodsWith this aim, data from 87 Standard bred racehorses which underwent a complete diagnostic evaluation for poor performance was reviewed. During gastroscopic examination, a 0-4 score was assigned to ESGD, while EGGD was evaluated for absence/presence; a total EGUS score was obtained by adding 1 point to ESGD score in horses showing concomitant EGGD. Fitness parameters obtained during incremental treadmill test included speed at a heart rate of 200 bpm (V200), speed and heart rate at a blood lactate of 4 mmol/L (VLa4, HRLa4), peak lactate, lactate and heart rate at 30 minutes post-exercise, maximum speed, minimum pH and maximum hematocrit. The associations between fitness parameters and EGUS and ESGD scores were evaluated by Spearman correlation, while Mann-Whitney test was used to compare them between horses with or without EGGD. Statistical significance was set at p<0.05.ResultsEGUS grade was inversely correlated with V200 (p = 0.0025) and minimum pH (p = 0.0469); ESGD grade was inversely correlated with V200 (p = 0.0025) and VLa4 (p = 0.0363). Although a trend was observed, no significant differences in V200 were observed between horses with or without EGGD (p = 0.073); horses with EGGD reached a lower minimum pH (p = 0.0087).DiscussionThese results show a negative association between aerobic capacity and EGUS, in particular ESGD. Although different hypotheses have been proposed, including abdominal pain and decreased appetite due to lactate accumulation, the underlying mechanisms are still unknown, and it is not clear whether EGUS represents a cause or a consequence of an early lactate accumulation and post-exercise acidosis.

Increased adrenocortical response to adrenocorticotropic hormone (ACTH) in sport horses with equine glandular gastric disease (EGGD)

Simple SummaryEquine gastric ulcer syndrome (EGUS) is a highly prevalent disease with a major clinical importance due to its negative effects on the welfare and performance of horses. EGUS can be distinguished into two different diseases depending on which anatomical region is affected: equine glandular gastric disease (EGGD), in which there is a lesion in the glandular stomach, and equine squamous gastric disease (ESGD), in which the alteration appears in the non-glandular stomach. EGUS has nonspecific clinical signs, and its underlying mechanism has not been completely elicited. Therefore, it would be of interest to clarify the pathophysiology and identify potential biomarkers of this syndrome. This study detected differences in the salivary and serum proteome between horses with EGUS and healthy horses, and also between horses with EGGD and ESGD. The most upregulated proteins in EGGD were related to the immune activation whereas, in horses with ESGD, the proteins with the most significant changes were associated with the squamous cell regulation and growth. Compared to serum, saliva had a higher number of proteins showing significant changes and also showed a different pattern of changes, indicating that the proteins in both fluids show a different response to the disease and can provide complementary information.Changes in the salivary proteome in 12 horses with the two diseases included in equine gastric ulcer syndrome (EGUS), equine glandular gastric disease (EGGD) (n = 6) and equine squamous gastric disease (ESGD) (n = 6), were evaluated using a high-resolution LC-MS/MS analysis of TMT-labelled peptides and compared to 10 healthy control horses. Serum was also analysed for comparative purposes. The comparison between the horses with EGGD and controls showed significant changes in 10 salivary proteins, whereas 36 salivary proteins were differently abundant between ESGD and control groups. The most upregulated proteins in the case of EGGD were related to immune activation whereas, in horses with ESGD, the most significantly changed proteins were associated with squamous cell regulation and growth. Compared to serum, saliva showed a higher number of proteins with significant changes and a different pattern of changes. The proteins identified in our study, in addition to providing new information about the pathophysiological mechanisms in these diseases, could have the potential to be novel biomarkers for the diagnosis or monitoring of EGGD and ESGD.

Equine Gastric Ulcers Syndrome (EGUS) is a worldwide disease present in equids of different breeds, activity levels, and age groups. It is divided into two different illnesses: Equine Squamous Gastric Disease (ESGD) affecting the squamous mucosa and Equine Glandular Gastric Disease (EGGD) affecting the glandular mucosa. The historical horserace is a traditional competition that is common in Italy. They can be divided into two different types: speed races and jousting tournaments. Anglo-Arabians and Thoroughbreds are used for the two competitions with training and management systems similar to those used in races performed on a racetrack. The aim of this study was to determine the prevalence of ESGD and EGGD in horses used for historical horseracing and evaluate the differences in the presence of the diseases in animals of the two different breeds used for the two types of competition. A cohort of 73 horses was enrolled in this study, which were stabled in 10 training facilities and performed two jousting tournaments and one speed race. An ESGD at least of grade 2 was found in 88% of horses with all degrees of severity seen; EGGD was diagnosed in 45% of animals. In this cohort of horses, the presence and severity of ESGD and EGGD are similar to that in reports in racehorses performing on racetracks. Anglo-Arabians used for speed races are more affected by ESGD and EGGD, which is probably because they are involved in a higher number of races and travel more during the year compared to Thoroughbreds used for jousting competitions.

SummaryThe study was performed on 39 live donkeys that underwent gastroscopic examination. The lesions were recorded in accordance with the European College of Equine Internal Medicine Consensus Statement guidelines. The presence of Gasterophilus sp. larvae was also recorded. Larvae were collected and identified to species level. Fisher's exact test was used to compare different prevalence values for sex, age, and anatomical distribution of lesions. Gastric lesions were present in 20/39 (51.3% [35.6–67%]) donkeys; 19/39 (48.7% [95% confidence interval = 33–64.4%]) were affected only by equine squamous gastric disease (ESGD), while 1/39 (2.6% [0–7.5%]) showed both ESGD and equine glandular gastric disease (EGGD), thus 95% of positive donkeys showed lesions located in the nonglandular mucosa. The ESGD grade was 0/4 (48.7% [33–64.4%]) in 19/39, 1/4 (12.8% [2.3–23.3%]) in 5/39, 2/4 (25.6% [11.9–39.5%]) in 10/39, 3/4 in 4/39 (10.3% [0.7–19.8%]) and 4/4 in 1/39 (2.6% [0–7.5%]) donkeys, respectively. The EGGD lesion was a mild depression in the ventral glandular fundus. ESGD was primary in all the donkeys included and lesions were located around the cardia and along the lesser curvature. Gasterophilus sp. larvae were present in all animals and were identified as third‐stage larvae of Gasterophilus intestinalis. No animals showed clinical signs of equine gastric ulcer syndrome (EGUS). No significant differences relating to sex, age or breed were found in the prevalence of EGUS in this study, while the proportion of donkeys affected by ESGD was statistically higher than those affected by EGGD. To the best of our knowledge, this is the first report on the gastroscopic evaluation of EGUS in live donkeys. Our results show a higher prevalence of EGUS in live donkeys than values previously reported by other authors in donkeys that were dead or had been subjected to euthanasia. The detection of third‐stage G. intestinalis larvae was not unexpected since these can be found in the stomach of equids throughout the year, and G. intestinalis has been reported as the most common Gasterophilus sp. in Italy.