The usability of peritoneal lactate concentration as a prognostic marker in horses with severe colic admitted to a veterinary teaching hospital

Colic can be a life‐threatening condition in horses and there is a need for parameters that can help determine the prognosis and need for surgery. The aim of the study was to investigate whether peritoneal fluid (PF) lactate concentration is useful for this purpose in horses with severe colic presented to a veterinary hospital. During a 16 month period, the peritoneal fluid (PF) lactate concentration was measured in 74 of 760 colic horses admitted to the Utrecht University equine clinic using a portable analyser. When comparing survivors and nonsurvivors, heart rate, PF and blood lactate concentrations and blood glucose concentration were significantly higher in horses that did not survive. No horse with a PF lactate concentration >9.4 mmol/l survived. The presence of a strangulating lesion was also significantly associated with nonsurvival, as was PF colour: no horse with red PF survived in the present series. In horses with yellow PF, the blood glucose concentration was correlated with the presence of a strangulating intestinal lesion. Peritoneal lactate concentrations can be easily and rapidly measured using a portable analyser and may be useful in assessing the prognosis and/or need for surgery in equine colic cases.     

Use of the Accusport semi-automated analyser to determine blood lactate as an aid in the clinical assessment of horses with colic

The most useful diagnostic methods in the initial evaluation of horses with colic assess the morphological and functional status of the gastrointestinal tract and cardiovascular status. This evaluation is best achieved using a combination of clinical and laboratory data. Blood lactate concentration (BL) is one of these variables. BL rises mainly due to poor tissue perfusion and anaerobic glycolysis associated with shock, providing an indicator of both the severity of disease and its prognosis. A hand-held lactate meter, Accusport, provides a rapid (60 seconds), inexpensive dry-chemical-based determination of BL. This trial evaluated the Accusport's ability to provide BL data as an adjunct to the initial clinical evaluation of horses with colic. The accuracy of the Accusport was tested by evaluation of its interchangeability with the benchmark enzymatic kit evaluation of BL in a trial using data collected firstly from 10 clinically normal control horses and subsequently from 48 horses presented with signs of colic. The BL values were recorded together with the clinical variables of heart rate (HR), capillary refill time (CRT), haematocrit (Hct), and pain character and severity on the initial assessment of the colic horses. Information regarding choice of therapeutic management (medical or surgical) and eventual case outcome (full recovery or died/euthanased) was recorded. The Accusport was found to be interchangeable with the enzymatic kit for recording BL values in colic horses with BL < 10 mmol/l, which is within the BL range associated with survival. The interchangeability of an additional, laboratory-based wet chemical assay for BL, the Stat 7 was simultaneously evaluated for the colic and control horses. The Stat 7 was found to be interchangeable with the enzymatic kit for BL determination of colic horses. No linear associations between BL values with HR, CRT, Hct or pain assessment were observed. No relationship with either selection of therapeutic method or eventual case outcome was observed. All horses with BL >8 mmol/l died or were euthanased.     

Determination of the source of increased serotonin (5-HT) concentrations in blood and peritoneal fluid of colic horses with compromised bowel

REASONS FOR PERFORMING STUDY: Increased plasma (5-HT) concentrations are reported in horses predisposed to develop laminitis and after i.v. infusion of endotoxins. In the equine jejunum contractile 5-HT1A-like receptors show tachyphylaxia upon prolonged activation with 5-HT. Therefore, increased systemic 5-HT release in colic horses could play a possible role in the pathophysiology of ileus. OBJECTIVE: To investigate possible increased systemic release of 5-HT in colic horses with compromised bowel and to identify the source of 5-HT overload. METHODS: Concentrations of 5-HT were determined in plasma and peritoneal fluid (PF) of healthy horses (n = 10), strangulating small intestinal colic horses (n = 18), nonsurgical colic horses (n = 10) and cryptorchid stallions (n = 6). It was attempted to identify the source of 5-HT overload by comparing the blood and PF 5-HT concentrations within horses and by assessing the in vivo platelet activation through determination of the beta-thromboglobulin (beta-TG)/platelet factor 4 (PF4) ratio. RESULTS: All horses in the strangulating small intestinal colic group had plasma (P = 0.006) and PF (P = 0.01) 5-HT concentrations above those found in the control group. Plasma beta-TG/PF4 ratio in these horses exceeded 2 in all cases, indicating in vivo platelet activation. Concentrations of 5-HT in PF of colic horses with compromised bowel were significantly lower than the corresponding plasma concentrations (P = 0.005). Potential relevance: In horses with compromised bowel, significant amounts of 5-HT can be released into the systemic circulation, through massive release of platelet-stored 5-HT. 5-HT is a very potent proinflammatory, vasoconstrictive and immunomodulatory agent. In view of the rapid and prolonged tachyphylaxia, shown for the jejunal 5-HT1A-like receptors, this increased systemic 5-HT release could play a role in the pathophysiology of ileus in horses.     

Evaluation of plasma catecholamine and serum cortisol concentrations in horses with colic

OBJECTIVE: To evaluate plasma epinephrine and norepinephrine concentrations and serum cortisol concentration in horses with colic and assess the relationship of these variables with clinical signs, routinely measured clinicopathologic variables, and outcome in affected horses. DESIGN: Prospective observational study. ANIMALS: 35 horses with colic. PROCEDURE: Blood samples were collected within 30 minutes of arrival at the veterinary hospital from horses referred because of colic. Plasma and serum samples were analyzed for cortisol, epinephrine, norepinephrine, lactate, and electrolyte concentrations and acid-base variables. Heart rate at admission and outcome (survival or nonsurvival) were recorded. Univariate logistic regression was used to calculate crude (unadjusted) odds ratios and 95% confidence intervals. RESULTS: Of the 35 horses with colic, 26 survived. Higher plasma epinephrine, plasma lactate, and serum cortisol concentrations were significantly associated with increased risk of nonsurvival, but plasma norepinephrine concentration was not associated with outcome. Plasma epinephrine concentration was significantly correlated with heart rate (r = 0.68), plasma lactate concentration (r = 0.87), blood pH (r = -0.83), anion gap (r = 0.74), and base excess (r = -0.81). CONCLUSIONS AND CLINICAL RELEVANCE: The risk of death appears to be greater in colic-affected horses with high circulating concentrations of epinephrine and cortisol. The correlation of epinephrine with other biochemical markers of illness severity and with heart rate indicates that the degree of sympathetic activation in horses with colic can be inferred from routinely measured variables.     

Comparative biochemical analyses of venous blood and peritoneal fluid from horses with colic using a portable analyser and an in-house analyser

Fifty-six horses with colic were examined over a period of three months. The concentrations of glucose, lactate, sodium, potassium and chloride, and the pH of samples of blood and peritoneal fluid, were determined with a portable clinical analyser and with an in-house analyser and the results were compared. Compared with the in-house analyser, the portable analyser gave higher pH values for blood and peritoneal fluid with greater variability in the alkaline range, and lower pH values in the acidic range, lower concentrations of glucose in the range below 8.3 mmol/l, and lower concentrations of lactate in venous blood in the range below 5 mmol/l and in peritoneal fluid in the range below 2 mmol/l, with less variability. On average, the portable analyser underestimated the concentrations of lactate and glucose in peritoneal fluid in comparison with the in-house analyser. Its measurements of the concentrations of sodium and chloride in peritoneal fluid had a higher bias and were more variable than the measurements in venous blood, and its measurements of potassium in venous blood and peritoneal fluid had a smaller bias and less variability than the measurements made with the in-house analyser.     

Effects of aerobic and anaerobic fluid collection on biochemical analysis of peritoneal fluid in healthy horses and horses with colic

Objective: To determine whether in healthy horses and those with colic, exposure of peritoneal fluid to room air affects values obtained on biochemical analysis. Study Design: Prospective study. Animals: Adult horses with a primary complaint of acute abdominal pain (n=29) and 12 healthy horses. Methods: Peritoneal fluid was aseptically collected under aerobic and anaerobic conditions. After collection, pH, PCO₂, PO₂, HCO₃^?, Na⁺, ionized Ca²⁺, K⁺, lactate, and glucose were immediately measured using a commercial blood gas analyzer. Biochemical variables were compared between aerobically and anaerobically obtained samples using a paired t‐test. Results: In healthy horses, peritoneal fluid samples collected under anaerobic conditions had higher PCO₂ and ionized Ca²⁺ and lower PO₂, HCO₃^?, and pH compared with samples exposed to air. No differences were observed for K⁺, Na⁺, glucose, and lactate. In horses with colic, samples collected anaerobically had higher PCO₂, ionized Ca²⁺, Na⁺, and glucose and lower PO₂, HCO₃^?, and pH value compared with samples exposed to air. No differences were observed for K⁺ and lactate. Conclusion: Exposure of peritoneal fluid to room air had a significant effect on pH, PCO₂, PO₂, and variables associated or dependent on changes in pH such as HCO₃^? and ionized Ca²⁺. Interpretation of biochemical analysis of peritoneal fluid may be influenced by sample collection method.     

Association of admission plasma D-dimer concentration with diagnosis and outcome in horses with colic

Background: Coagulopathies detected in horses with gastrointestinal problems seem to be associated with poor outcome. Plasma D‐Dimer concentration is a sensitive test for assessing coagulopathies. Hypothesis: Plasma D‐Dimer concentration tested on admission is related to diagnosis and outcome in horses with colic. Animals: Four hundred and ninety three horses referred for evaluation of abdominal pain. Methods: Prospective observational clinical study. Horses were grouped according to diagnosis (medical and surgical intestinal obstructions, ischemic disorders with and without intestinal resection, enteritis, peritonitis), outcome (survivors, nonsurvivors), and number of coagulopathies (normal profile, 1 or 2 coagulopathies, subclinical disseminated intravascular coagulation [DIC]). Blood samples were collected on admission and plasma D‐Dimer concentration, clotting times (PT and aPTT), and antithrombin activity were determined. Positive likelihood ratios (LR+) were calculated for evaluation of D‐Dimer cut‐off values, which were later tested in a logistic regression model. Results: Horses with enteritis or peritonitis had significantly (P < .001) higher plasma D‐Dimer concentrations and more severe coagulopathies on admission than horses with other diagnoses. Nonsurvivors also had significantly (P < .001) higher plasma D‐Dimer concentrations at presentation than did survivors, and those horses with subclinical DIC on presentation had an odds ratio (OR) 8.6 (95% confidence interval [CI], 3.3–22.5, P < .001) for nonsurvival. Finally, D‐Dimer concentrations >4,000 ng/mL had a LR+ of 5.9 and an OR 8.8 (95% CI, 4.5–17.1, P < .001) for nonsurvival. Conclusion and Clinical Importance: Plasma D‐Dimer concentration measured on admission can be used to facilitate diagnosis and outcome prediction in horses with colic. A potential cut‐off value for nonsurvival was found at approximately 4,000 ng/mL.     

Peritoneal concentrations of transforming growth factor beta in horses with colic

Reasons for performing the study: In man, peritoneal transforming growth factor beta (TGF‐β) is associated with peritoneal diseases and subsequent adhesion formation. No studies on plasma and peritoneal TGF‐β concentrations in horses with colic are available. Objectives: 1) To determine both plasma and peritoneal TGF‐β₁ and TGF‐β₃ concentrations in horses with different types of colic (not previously subjected to abdominal surgery); 2) to compare these concentrations according to the type of peritoneal fluid (transudate, modified transudate and exudate); and 3) to compare and correlate plasma and peritoneal concentrations of TGF‐β₁ and TGF‐β₃ and the types of peritoneal fluid according to the colic group and outcome. Methods: Peritoneal fluid and plasma samples from 78 horses with colic and 8 healthy horses were obtained. Patients were classified according to diagnosis (obstructions, enteritis, ischaemic disorders and peritonitis), peritoneal fluid analysis (transudate, modified transudate and exudate), and outcome (survivors and nonsurvivors). Plasma and peritoneal TGF‐β₁ and TGF‐β₃ concentrations were determined by ELISA. Data were analysed by parametric and nonparametric tests. P≤0.05 was considered as statistically significant. Results: Concentrations of peritoneal fluid TGF‐β₁ were significantly (P = 0.01) higher in horses with peritonitis in comparison with all other colic groups and controls. Horses with ischaemic lesions had significantly (P = 0.01) higher concentrations of peritoneal TGF‐β₁ in comparison with controls and the group of horses with obstructions. Peritoneal TGF‐β₁ concentration also was significantly (P = 0.01) higher in exudates in comparison with transudates. Peritoneal TGF‐β₁ and TGF‐β₃ concentrations and plasma TGF‐β₁ concentration were significantly increased in nonsurvivors compared to survivors (P = 0.001, P = 0.004 and P = 0.05, respectively). Conclusions: Peritoneal TGF‐β₁ concentration was higher in horses with severe gastrointestinal diseases (ischaemic intestinal lesions and peritonitis), in horses with an altered peritoneal fluid (exudate), and in nonsurvivors. Potential relevance: Peritoneal TGF‐β concentration increases in horses with severe gastrointestinal disease as an anti‐inflammatory response.     

Myeloperoxidase assay in plasma and peritoneal fluid of horses with gastrointestinal disease

Gastrointestinal disorders, especially strangulating intestinal obstructions, are still a major cause of illness and death in the horse. Circulating lipopolysaccharides may activate both neutrophils and monocytes. The activated neutrophils release myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to evaluate MPO concentrations in the plasma and peritoneal fluid (PF) of horses with colic and to check the hypothesis that these concentrations would be higher in a case of strangulating obstruction than in cases of nonstrangulating disease. By using a specific enzyme-linked immunosorbent assay for equine MPO, we determined the MPO concentrations in horses admitted to a clinic for colic. Horses with nonstrangulating or strangulating obstruction of the large intestine (NSLI or SLI), strangulating obstruction of the small intestine (SSI), or inflammatory bowel disease (IBD) were compared with healthy horses. The horses with SLI, SSI, or IBD had significantly higher MPO levels in plasma and PF than did those in the other 2 groups. The mean plasma level was significantly higher in the horses with NSLI than in the healthy horses. High MPO values in PF indicated necrotic bowel. These results show that neutrophil activation occurs during nonstrangulating and strangulating intestinal obstruction in horses and that the plasma and PF MPO concentrations may be a marker of the severity of the disease.     

Determination of grapiprant plasma and urine concentrations in horses

ObjectiveNon-steroidal anti-inflammatory drugs are inhibitors of cyclooxygenase (COX) in tissues and used as therapeutic agents in different species. Grapiprant, a member of the piprant class of compounds, antagonizes prostaglandin receptors. It is a highly selective EP4 prostaglandin E₂ receptor inhibitor, thereby limiting the potential for adverse effects caused by wider COX inhibition. The objectives of this study were to determine if the approved canine dose would result in measurable concentrations in horses, and to validate a chromatographic method of analysis for grapiprant in urine and plasma.Study designExperimental study.AnimalsA total of six healthy, adult mixed-breed mares weighing 502 ± 66 (397–600) kg and aged 14.8 ± 5.3 (6–21) years.MethodsMares were administered one dose of 2 mg kg^–1 grapiprant via nasogastric tube. Blood and urine samples were collected prior to and up to 48 hours after drug administration. Drug concentrations were measured using high-performance liquid chromatography.ResultsGrapiprant plasma concentrations ranged from 71 to 149 ng mL^–1 with the mean peak concentration (106 ng mL^–1) occurring at 30 minutes. Concentrations were below the lower limit of quantification (50 ng mL^–1) in four of six horses at 1 hour and in all six horses by 2 hours after drug administration. Grapiprant urine concentrations ranged from 40 to 4077 ng mL^–1 and were still detectable at 48 hours after administration.Conclusions and clinical relevanceCurrently, there are no published studies looking at the pharmacodynamics of grapiprant in horses. The effective concentration needed to control pain in dogs ranges 114–164 ng mL^–1. Oral administration of grapiprant (2 mg kg^–1) in horses did not achieve those concentrations. The dose was well tolerated; therefore, studies with larger doses could be conducted.     

Evaluation of cerebrospinal fluid lactate and plasma lactate concentrations in anesthetized dogs with and without intracranial disease

The objectives of this study were to establish a reference interval for canine cerebrospinal fluid lactate (CSFL) and to compare CSFL and plasma lactate (PL) concentrations in anesthetized dogs with and without intracranial disease. Using a prospective study, canine blood and cerebrospinal fluid were collected for lactate analysis in 11 dogs with intracranial disease after undergoing magnetic resonance imaging (MRI) (Group ID-MRI), in 10 healthy dogs post-MRI (Group H-MRI), and in 39 healthy dogs after induction of anesthesia (Group H-Sx). Dogs were anesthetized for the procedures using different anesthetic protocols. Neurological scores (NS) and sedation scores (SS) were assessed pre-anesthesia in ID-MRI dogs. The CSFL reference interval [90% confidence interval (CI) for lower and upper limits] was 1.1 (1.0 to 1.2) to 2.0 (2.0 to 2.1) mmol/L. Mean ± SD CSFL concentrations were: ID-MRI, 2.1 ± 0.8; H-MRI, 1.6 ± 0.4; and H-Sx, 1.6 ± 0.2 mmol/L. There was a tendency for higher CSFL in dogs in the ID-MRI group than in those in the H-MRI or H-Sx groups (P = 0.12). There was agreement between CSFL and PL in ID-MRI dogs (P = 0.007), but not in dogs in H-MRI (P = 0.5) or H-Sx (P = 0.2). Of the ID-MRI dogs, those with worse NS had higher CSFL (r² = 0.44). The correlation between CSFL and PL in dogs with intracranial disease and between worse NS and higher CSFL warrants further investigation into the use of CSFL and PL for diagnostic and prognostic purposes.     

The diagnostic and prognostic value of alkaline phosphatase activity in serum and peritoneal fluid from horses with acute colic

Alkaline phosphatase (ALP) is an enzyme present in intestinal mucosa, bile, bone, and renal tubule cells. We sought to assess the diagnostic and prognostic relationships of total ALP (ALPt) activity and that of intestine-derived ALP (ALPi) in serum and peritoneal fluid of 126 horses with colic. ALPt and ALPi activities were measured in both serum and peritoneal fluid by using both standard and L-phenylalanine-based buffers, respectively. Neither ALPt nor ALPi activity were useful in classifying type or severity of intestinal damage. ALPt and ALPi activities in peritoneal fluid were lowest in horses suffering from simple medical colic (39 international units [U]/L [19-60 U/L]; versus 31 U/L [16-44 U/L], median [interquartile range], P < .001) and nonstrangulated surgical lesions (45 U/L [30-62 U/L] versus 36 U/L [23-54 U/L], P < .001), and highest in surgical cases with suspected ulceration (109 U/L [60-1,113 U/L] versus 83 U/L [52-970 U/L], P < .001), strangulation (114 U/L [69-240 U/L] versus 94 U/L [56-191 U/L], P < .001), peritonitis (313 U/L [110-2,227 U/L] versus 283 U/L [91-1,800 U/L], P < .001) or intestinal rupture (687 U/L [205-852 U/L] versus 564 U/L [166-732 U/L], P < .001). Higher ALPt and ALPi activities in peritoneal fluid were associated with greater intestinal damage, increased probability of surgery, and a worse prognosis. The use of L-phenylalanine buffer in both serum and peritoneal fluid did not improve the sensitivity of the test. Based on these results, total ALP activity in peritoneal fluid may help in identifying ischemic or inflammatory bowel lesions in horses with acute colic.