Relationship between the Cycle Threshold Value (Ct) of a Salmonella spp. qPCR Performed on Feces and Clinical Signs and Outcome in Horses.

The objective of this retrospective study was to evaluate the clinical significance of fecal quantitative real-time polymerase chain reaction (qPCR) Salmonella results when taking the cycle threshold values (Ct) into account. The study included 120 Salmonella qPCR-positive fecal samples obtained from 88 hospitalized horses over a 2-year period. The mean Ct of the qPCR test was evaluated in regard to (1) clinical outcome and (2) systemic inflammatory response syndrome (SIRS) status (no SIRS, moderate SIRS, or severe SIRS) of the sampled horses. An ROC analysis was performed to establish the optimal cut-off Ct values associated with severe SIRS. The mean ± SD Ct value was significantly lower in samples (1) from horses with a fatal issue (27.87 ± 5.15 cycles) than in surviving horses (31.75 ± 3.60 cycles), and (2) from horses with severe SIRS (27.87 ± 2.78 cycles) than from horses with no (32.51 ± 3.59 cycles) or moderate (31.54 ± 3.02 cycles) SIRS. In the ROC analysis, the optimal cut-off value of Ct associated with a severe SIRS was 30.40 cycles, with an AUC value of 0.84 [95% confidence interval 0.76-0.91] and an OR of 0.64 [0.51-0.79]. Results suggest that including the Ct value in the interpretation of fecal qPCR results could improve the diagnostic value of this test for clinical salmonellosis in horses.

Evaluation of Bacterial Composition and Viability of Equine Feces after Processing for Transplantation.

Fecal microbiota transplantation (FMT) has been used empirically for decades in equine medicine to treat intestinal dysbiosis but evidence-based information is scarce. This in vitro study aimed at assessing the effect of a commonly used pre-FMT processing method on the bacterial composition and viability of the fecal filtrate. Three samples of fresh equine manure (T0) were processed identically: the initial manure was mixed with 1 L of lukewarm water and chopped using an immersion blender to obtain a mixture (T1), which was left uncovered during 30 min (T2) and percolated through a sieve to obtain a fecal filtrate (T3). Samples were taken throughout the procedure (Tn) and immediately stored at 4 °C until processing. The 16S rDNA amplicon profiling associated with propidium monoazide treatment was performed on each sample to select live bacteria. Analyses of α and ß diversity and main bacterial populations and quantitative (qPCR) analysis were performed and statistically compared (significance p < 0.05) between time points (T0-T3). No significant differences in ecological indices or mean estimated total living bacteria were found in the final fecal filtrate (T3) in regard to the original manure (T0); however, relative abundances of some minor genera (Fibrobacter, WCHB1-41_ge and Akkermansia) were significantly different in the final filtrate. In conclusion, the results support the viability of the major bacterial populations in equine feces when using the described pre-FMT protocol.

Analytical Performance Evaluation of the New GEM® Premier™ 5000 in Comparison to the Epoc® Blood Gas Analyzer in Horses.

Different blood gas analyzers are used in equine practice. Every machine needs to be validated, as they have not been designed for use in horses. The aim of this study was to compare the newly marketed GEM5000 machine to the formerly validated epoc machine for blood gas analysis in horses. In this prospective, non-blinded, comparative laboratory analyzer study, 43 equine blood samples were analyzed on both analyzers and values were compared between the two machines via Lin's concordance analysis, Passing-Bablok regression analysis and Bland-Altman plots. Duplicate measurements were conducted on the GEM5000 machine to evaluate precision. The GEM5000 failed to achieve the required precision for tHb, Hct and iCa2+, but achieved acceptable precision for all other parameters. Concordance correlation analysis revealed poor correlation for Na+, Cl-, iCa2+, K+, Hct and tHb, while there was an at least moderate agreement for all other parameters. Passing-Bablok regression revealed significant constant bias for pCO2, pO2, Cl-, and iCa2+ and significant proportional bias for pCO2, iCa2+ and SO2. Bland-Altman analysis revealed significant systematic bias for Na+, Cl-, iCa2+, K+, Hct, tHb and SO2. This study shows that while precision of the GEM5000 is good, values should not be used interchangeably with data obtained from other blood gas analyzers.

Comparison of Fecal Microbiota of Horses Suffering from Atypical Myopathy and Healthy Co-Grazers.

Equine atypical myopathy (AM) is caused by hypoglycin A (HGA) and methylenecyclopropylglycine (MCPG) intoxication resulting from the ingestion of seeds or seedlings of some Acer tree species. Interestingly, not all horses pasturing in the same toxic environment develop signs of the disease. In other species, it has been shown that the intestinal microbiota has an impact on digestion, metabolism, immune stimulation and protection from disease. The objective of this study was to characterize and compare fecal microbiota of horses suffering from AM and healthy co-grazers. Furthermore, potential differences in fecal microbiota regarding the outcome of diseased animals were assessed. This prospective observational study included 59 horses with AM (29 survivors and 30 non-survivors) referred to three Belgian equine hospitals and 26 clinically healthy co-grazers simultaneously sharing contaminated pastures during spring and autumn outbreak periods. Fresh fecal samples (rectal or within 30 min of defecation) were obtained from all horses and bacterial taxonomy profiling obtained by 16S amplicon sequencing was used to identify differentially distributed bacterial taxa between AM-affected horses and healthy co-grazers. Fecal microbial diversity and evenness were significantly (p < 0.001) higher in AM-affected horses as compared with their non-affected co-grazers. The relative abundance of families Ruminococcaceae, Christensenellaceae and Akkermansiaceae were higher (p ≤ 0.001) whereas those of the Lachnospiraceae (p = 0.0053), Bacteroidales (p < 0.0001) and Clostridiales (p = 0.0402) were lower in horses with AM, especially in those with a poor prognosis. While significant shifts were observed, it is still unclear whether they result from the disease or might be involved in the onset of disease pathogenesis.

High Seroprevalence Against Lawsonia intracellularis Among Adult Horses in Belgium.

Lawsonia intracellularis (LI) is an obligate intracellular gram-negative rod causing equine proliferative enteropathy (EPE). Occasional cases of EPE have been reported in foals living in Belgium, but the seroprevalence of equine LI in this country is unknown. The target population included clinically healthy adult horses, whose blood samples were collected and analyzed for specific IgG antibodies against LI using a blocking enzyme-linked immunosorbent assay test. The results were expressed as percentage of inhibition (PI). Samples that had a PI <20% were judged as negative, those between 20 and 30% as inconclusive, and those >30% were considered positive. A total of 356 blood samples were analyzed with 352 horses (98.8%) testing positive, 2 horses (0.6%) testing negative, and 2 horses (0.6%) showing inconclusive results. The large percentage of seropositive samples obtained in this study confirms a widespread exposure of Belgian horses to LI.

Effect of oral administration of omeprazole on the microbiota of the gastric glandular mucosa and feces of healthy horses.

BACKGROUND: Omeprazole administration is associated with changes in gastric and fecal microbiota and increased incidence of Clostridioides difficile enterocolitis in humans and dogs. HYPOTHESIS/OBJECTIVES: Study purpose was to assess the effect of omeprazole on gastric glandular and fecal microbiota in healthy adult horses. ANIMALS: Eight healthy horses stabled on straw and fed 100% haylage. METHODS: Prospective controlled study. Transendoscopic gastric glandular biopsies, gastric fluid, and fecal samples were obtained from each horse twice at a 7-day interval before the administration of omeprazole. Samples were taken on the same horses before and after a 7-day administration of omeprazole (4 mg/kg PO q24h). pH was assessed on fresh gastric fluid and other samples were kept at -20°C until analysis. Bacterial taxonomy profiling was obtained by V1V3 16S amplicon sequencing from feces and gastric glandular biopsies. Analysis of alpha, beta diversity, and comparison between time points were performed with MOTHUR and results were considered significant when P < .05. RESULTS: Gastric pH increased significantly after 7 days of omeprazole administration (P = .006). Omeprazole did not induce significant major changes in composition of fecal or gastric glandular microbiota, however, after administration, certain microbial genera became more predominant in the gastric glandular mucosa (lower Simpson's evenness, P = .05). Only the genus Clostridium sensu strictu_1 had a significant shift in the glandular gastric mucosa after omeprazole administration (P = .002). No population shifts were observed in feces. CONCLUSIONS AND CLINICAL IMPORTANCE: Oral administration of omeprazole could have fewer effects in gastrointestinal microbiota in the horse compared to other species.

Evaluation of an oral direct factor Xa inhibitor anticoagulant in healthy adult horses.

OBJECTIVE: To assess whether an oral direct factor Xa inhibitor (DiXaI) anticoagulant drug used at the low end of the recommended dose in people achieves presumed prophylactic plasma concentrations and does not induce bleeding in horses. DESIGN: Experimental study. SETTING: Field study. ANIMALS: Ten healthy adult horses. INTERVENTIONS: A DiXaI was administered at a dose of 0.125 mg/kg every 24 h orally for 4 days. Following a wash-out period of 2 weeks, 8 of 10 horses received daily subcutaneous doses of a low molecular weight heparin (dalteparin) for 4 consecutive days at 50 IU/kg. In both trials, antifactor Xa activity was measured at baseline time and 3 hours after each dose administration. Activated partial thromboplastin time, prothrombin time, hematocrit, erythrocyte agglutination, and platelet aggregation were monitored throughout the study. In addition, an in vitro spiking experiment was performed to demonstrate anticoagulant activity of this DiXaI in horse plasma. MAIN RESULTS: When treated with the DiXaI, this group of horses did not achieve the suggested thromboprophylactic plasma range of antifactor Xa activity (0.1-0.2 IU/mL), except for 1 horse after the first administration of the drug. In contrast, median values of plasma antifactor Xa activity 3 hours after receiving dalteparin were within the prophylactic range (0.16 IU/mL). No hemorrhagic events or erythrocyte agglutination were observed. In vitro addition of this DiXaI caused a concentration-dependent effect in antifactor Xa activity. CONCLUSIONS: At the low end of the recommended dose in people this oral formulation of DiXaI did not reach prophylactic plasma antifactor Xa activity in this group of healthy adult horses. Further studies are warranted in order to establish the prophylactic dose for horses.

Progression of plasma D-dimer concentration and coagulopathies during hospitalization in horses with colic.

OBJECTIVE: To assess the progression of plasma D-dimer concentrations and coagulation status in horses with different types of colic. DESIGN: Prospective clinical observational study performed between March 2004 and September 2008. SETTING: Veterinary university teaching hospital. ANIMALS: Horses admitted and treated for colic and hospitalized for >48 hours were considered. Animals were classified by diagnosis into medical obstructive conditions (MO), surgical obstructive conditions (SO), inflammatory conditions, and ischemic lesions (IS). INTERVENTIONS: Three blood samples were obtained from each horse (admission, at 24-48 h [or after surgery] and upon discharge). For each sample, plasma D-dimer concentration, prothrombin time, activated partial thromboplastin time, antithrombin activity, and the presence of subclinical disseminated intravascular coagulation were determined. MEASUREMENTS AND MAIN RESULTS: When median plasma D-dimer concentration values at admission and after 24-48 hours were compared, they were different but stable in horses with MO (1.29-1.95 nmol/L) and inflammatory conditions (5.70-6.69 nmol/L). However, 10-fold and 5-fold increases were observed, respectively, in SO (2.08 to 16.38 nmol/L) and IS (3.08 to 15.91 nmol/L) in the postoperative period. By 24-48 hours, the percentage of horses with coagulopathy increased in most groups (MO, 43 to 58%; SO, 50 to 96%, IS, 53 to 90%). By the time of discharge, 87% of horses with SO problems and 89% of horses with IS still had some form of coagulopathy documented. CONCLUSIONS: Throughout hospitalization, horses with MO problems had less severe coagulopathy and lower plasmatic D-dimer concentrations compared to other groups of horses. On admission, most horses with inflammatory conditions presented with coagulopathy. At 24-48 hours of hospitalization and following surgery, the hemostatic profile can differ markedly when compared to admission values.

Parallel testing of plasma iron and fibrinogen concentrations to detect systemic inflammation in hospitalized horses.

OBJECTIVES: To determine if plasma iron concentration is different between horses with and without systemic inflammation (SI) and to assess the accuracy for the detection of SI by assaying plasma iron and fibrinogen concentrations, individually or combined. To assess the prognostic value of plasma iron concentration and to describe the progression of plasma iron and fibrinogen concentrations during hospital follow-up, and its relation to SI and survival. DESIGN: Prospective observational study evaluating plasma iron and fibrinogen. SETTING: University veterinary teaching hospital. ANIMALS: Equine patients greater than 30 days of age. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Plasma iron and fibrinogen concentration was prospectively determined in hospitalized horses. Horses were classified into 2 groups: SI and non-SI. Horses were also classified according to clinical outcome. A group of control healthy horses was also included. A total of 135 horses were included in the study. Plasma iron concentration was significantly lower and fibrinogen concentration was higher in the SI group. Nonsurvivors had a mean plasma fibrinogen concentration significantly higher than survivors. The combination of plasma iron and fibrinogen has a high degree of specificity, sensitivity, and accuracy for the detection of SI in horses. Follow-up measurements were obtained in 48 horses. Surviving horses normalized plasma iron concentration during follow-up examination whereas nonsurviving horses had persistently low plasma iron concentrations. CONCLUSIONS: Plasma iron concentration alone is an accurate marker of SI in hospitalized horses. Alteration of both plasma iron and fibrinogen concentrations improves the specificity and positive predictive value for diagnosis of SI. Alteration of either one of both increases sensitivity and negative predictive value. Surviving horses normalized plasma iron concentrations during follow-up period. The combination of plasma iron and fibrinogen concentrations may help in the detection of SI. Follow-up of plasma iron concentrations may provide useful prognostic information.

Palatal sclerotherapy for the treatment of intermittent dorsal displacement of the soft palate in 51 standardbred racehorses.

This retrospective study evaluated the efficacy and side effects of palatal sclerotherapy in standardbred racehorses suspected to have intermittent dorsal displacement of the soft palate (IDDSP). Fifty-one horses were treated with multiple endoscopically guided injections of 3% sodium tetradecyl sulfate in the soft palate. Two groups were identified: those that had respiratory noises during exercise (n = 27) and those that did not (n = 24). Treatment was well-tolerated. Furthermore, horses significantly reduced their racing times for the last 400 m compared with their times before treatment and even when their times were compared to the mean times for horses in the same race. In conclusion, palatal sclerotherapy appears to be a suitable alternative therapeutic option for horses suspected to have IDDSP.

Costochondral junction osteomyelitis in 3 septic foals.

The costochondral junction constitutes a potential site of infection in septic foals and it could be favored by thoracic trauma. Standard radiographs and ultrasonography are useful tools for diagnosis of this condition and ultrasound-guided needle aspiration could permit the definitive confirmation of infection.

Coagulopathies in horses with colic.

The most common coagulopathy in horses with colic is a hypercoagulable state associated with disseminated intravascular coagulation. The intensity of this coagulopathy depends on the severity and duration of the gastrointestinal lesion, with the ischemic and inflammatory problems and peritonitis being the most frequently affected by coagulopathies. Early initiation of prophylactic therapy significantly reduces the severe hypercoagulable state in horses with intestinal conditions which are recognized to be at high risk for disseminated intravascular coagulation. In addition to the systemic coagulopathy observed in horses with colic, a peritoneal coagulopathy independent from that occurring in blood has been observed, and its recognition and assessment may have clinical usefulness in the diagnosis of the gastrointestinal diseases and outcome.