Biomarkers of endothelial activation and inflammation in dogs with organ dysfunction secondary to sepsis.

Introduction: Alteration in endothelial function during sepsis is thought to play a key role in the progression of organ failure. We herein compared plasma concentrations of endothelial activation biomarkers vascular endothelial growth factor (VEGF), hyaluronan (HA), plasminogen activator inhibitor-1 (PAI-1) and von Willebrand factor (vWF), as well as inflammatory mediator concentrations (IL-6, IL-8, IL-10, C-reactive protein and monocyte chemoattractant protein-1) in dogs with sepsis to healthy dogs. Methods: This study was a multicenter observational clinical trial conducted at two university teaching hospitals from February 2016 until July 2017. The study included 18 client-owned dogs hospitalized with sepsis and at least one distant organ dysfunction, as well as 20 healthy dogs. Plasma biomarker concentrations were measured using ELISA. Severity of illness in dogs with sepsis was calculated using the 5-variable acute physiologic and laboratory evaluation (APPLEFAST) score. Biomarker concentrations were compared between septic and healthy dogs using linear models. Results: Septic peritonitis was the most frequent source of sepsis (11/18; 61%), followed by pneumonia (4/18; 22%). Ten dogs (56%) had only 1 organ dysfunction, whereas 3 dogs (17%) had 2, 3 (17%) had 3, 1 (6%) had 4 and 1 (6%) had 5 organ dysfunctions. The median APPLEFAST score in the septic dogs was 28.5 (Q1-Q3, 24-31). Mean plasma concentrations of all endothelial and inflammatory biomarkers, except vWF, were higher in the sepsis cohort than in controls. The mean endothelial biomarker concentrations in the septic cohort ranged from ~2.7-fold higher for HA (difference in means; 118.2 ng/mL, 95% credible limit; 44.5-221.7) to ~150-fold for VEGF (difference in means; 76.6 pg./mL, 95% credible limit; 33.0-143.4), compared to the healthy cohort. Fifteen dogs with sepsis (83%) died; 7 (46%) were euthanized and 8 (53%) died during hospitalization. Conclusion: Dogs with naturally occurring sepsis and organ dysfunction had higher mean concentrations of biomarkers of endothelial activation and inflammation compared to healthy dogs, broadening our understanding of the pathophysiology of sepsis secondary to endothelial dysfunction.

The Effects of Resuscitative Fluid Therapy on the Endothelial Surface Layer.

The goal of resuscitative fluid therapy is to rapidly expand circulating blood volume in order to restore tissue perfusion. Although this therapy often serves to improve macrohemodynamic parameters, it can be associated with adverse effects on the microcirculation and endothelium. The endothelial surface layer (ESL) provides a protective barrier over the endothelium and is important for regulating transvascular fluid movement, vasomotor tone, coagulation, and inflammation. Shedding or thinning of the ESL can promote interstitial edema and inflammation and may cause microcirculatory dysfunction. The pathophysiologic perturbations of critical illness and rapid, large-volume fluid therapy both cause shedding or thinning of the ESL. Research suggests that restricting the volume of crystalloid, or "clear" fluid, may preserve some ESL integrity and improve outcome based on animal experimental models and preliminary clinical trials in people. This narrative review critically evaluates the evidence for the detrimental effects of resuscitative fluid therapy on the ESL and provides suggestions for future research directions in this field.

Coagulation factor activity patterns of venom-induced consumption coagulopathy in naturally occurring tiger snake (Notechis scutatus) envenomed dogs treated with antivenom.

BACKGROUND: Venom-induced consumption coagulopathy (VICC) from tiger snake (Notechis scutatus) envenomation results in a dose-dependent coagulopathy that is detectable on coagulometry. However, individual coagulation factor activities in dogs with tiger snake envenomation have not been determined. This study aimed to characterise VICC and the time course of recovery in tiger snake envenomed dogs and to investigate an association between tiger snake venom (TSV) concentrations and factor activity. METHODS: This was a prospective, observational, cohort study. The study cohort was 11 dogs of any age, breed, sex, body weight >10 kg, confirmed serum TSV on ELISA and treated with antivenom. Blood was collected at enrolment before antivenom administration, then at 3, 12 and 24 h after antivenom administration. Tiger snake venom concentrations were detected with a sandwich ELISA. Fibrinogen was measured using a modified Clauss method, and coagulation factors (F) II, V, VII, VIII and X were measured with factor-deficient human plasma using a modified prothrombin (PT) and activated partial thromboplastin (aPTT) method. Linear mixed models, with multiple imputations of censored observations, were used to determine the effect of time and TSV concentration on the coagulation times and factor activity. This cohort was compared to 20 healthy controls. RESULTS: At enrolment, there were severe deficiencies in fibrinogen, FV and FVIII, with predicted recovery by 10.86, 11.75 and 13.14 h after antivenom, respectively. There were modest deficiencies in FX and FII, with predicted recovery by 20.57 and 32.49 h after antivenom, respectively. No changes were detected in FVII. Prothrombin time and aPTT were markedly prolonged with predicted recovery of aPTT by 12.58 h. Higher serum TSV concentrations were associated with greater deficiencies in FII, FV and FVIII, and greater prolongations in coagulation times. The median (range) serum TSV concentration was 57 (6-2295) ng/mL. CONCLUSIONS: In tiger snake envenomed dogs, we detected a profound, TSV-concentration-related consumption of select coagulation factors, that rapidly recovered toward normal. These findings allowed further insight into tiger snake VICC in dogs.

In vitro effects of lactated Ringer's solution, hypertonic saline, hydroxyethyl starch, hypertonic saline/hydroxyethyl starch, and mannitol on thromboelastographic variables of canine whole blood.

OBJECTIVE: To assess the in vitro effects of crystalloid and colloid IV fluids on the thromboelastographic (TEG) variables of canine whole blood. DESIGN: In vitro experimental study. SETTING: Veterinary teaching hospital. ANIMALS: Twenty-two healthy dogs. INTERVENTION: Citrated whole blood samples collected from healthy dogs were diluted with 3.4% hypertonic saline (HTS 3.4), 7% hypertonic saline (HTS 7), and 20% mannitol at 8% and 16% dilutions; hydroxyethyl starch 130/0.4 (HES 130/0.4) at 16% dilution; lactated Ringer's solution (LRS) at 16%, 33%, and 66% dilutions; and HTS 7-HES 130/0.4 at 25% and 50% dilutions. Kaolin-activated TEG analysis was concurrently performed on diluted and control (undiluted) samples. MEASUREMENTS AND MAIN RESULTS: Dilution of canine whole blood with LRS compared to control reduced α angle and MA at both 33% (P = 0.009 and P = 0.011, respectively) and 66% dilution (P < 0.001 and P < 0.001, respectively), and prolonged K time at 66% dilution (P = 0.003). At 16% dilution, HTS 3.4, prolonged R time (P = 0.007), while mannitol, a fluid iso osmolar to HTS 3.4, prolonged K time (P = 0.006), reduced α angle (P < 0.001), MA (P = 0.046), and LY60 (P = 0.015). At 8% dilution, HTS 7, a fluid of high osmolarity and tonicity, prolonged R time (P = 0.009) and reduced MA (P = 0.015), while all measured TEG variables were altered at the 16% dilution (P < 0.01 for all variables). HES 130/0.4 reduced α angle (P = 0.031) and MA (P = 0.001) and increased LY60 (P < 0.001) at 16% dilution. Comparing different fluid types, HES 130/0.4 and HTS 3.4 had no to minor, mannitol intermediate, and HTS 7 profound effects on TEG variables (P < 0.05) when compared to LRS at the same dilution. CONCLUSIONS: In vitro dilution of canine whole blood with commonly used IV fluids leads to thromboelastographic changes consistent with hypocoagulability in a dose dependent manner for all fluid types tested. Viscoelastic changes are also influenced by fluid characteristics, specifically tonicity and osmolarity.

The endothelial glycocalyx: Structure and function in health and critical illness.

OBJECTIVE: To conduct a narrative review of the current literature in reference to the structure and function of the endothelial glycocalyx (EG) and its contribution to the pathophysiology of conditions relevant to the veterinary emergency and critical care clinician. Novel therapies for restoring or preserving the EG will also be discussed. DATA SOURCES: Online databases (PubMed, CAB abstracts, Scopus) were searched between January 1st 2017 and May 1st 2017 for English language articles without publication date restriction. Keywords included EG, endothelial surface layer, degradation, syndecan-1, heparan sulfate, critical illness, sepsis, trauma, and therapeutics. DATA SYNTHESIS: The EG is a complex and important structure located on the luminal surface of all blood vessels throughout the body. It plays an important role in normal vascular homeostasis including control of fluid exchange across the vascular barrier. Loss or degradation of the EG has an impact on inflammation, coagulation, and vascular permeability and tone. These changes are essential components in the pathophysiology of many conditions including sepsis and trauma. A substantial body of experimental animal and human clinical research over the last decade has demonstrated increased circulating concentrations of EG degradation products in these conditions. However, veterinary-specific research into the EG and critical illness is currently lacking. The utility of EG degradation products as diagnostic and prognostic tools continues to be investigated and new therapies to preserve or improve EG structure and function are under development. CONCLUSIONS: The recognition of the presence of the EG has changed our understanding of transvascular fluid flux and the pathophysiology of many conditions of critical illness. The EG is an exciting target for novel therapeutics to improve morbidity and mortality in conditions such as sepsis and trauma.

Clinical use of plasma lactate concentration. Part 1: Physiology, pathophysiology, and measurement.

OBJECTIVE: To review the current literature with respect to the physiology, pathophysiology, and measurement of lactate. DATA SOURCES: Data were sourced from veterinary and human clinical trials, retrospective studies, experimental studies, and review articles. Articles were retrieved without date restrictions and were sourced primarily via PubMed, Scopus, and CAB Abstracts as well as by manual selection. HUMAN AND VETERINARY DATA SYNTHESIS: Lactate is an important energy storage molecule, the production of which preserves cellular energy production and mitigates the acidosis from ATP hydrolysis. Although the most common cause of hyperlactatemia is inadequate tissue oxygen delivery, hyperlactatemia can, and does occur in the face of apparently adequate oxygen supply. At a cellular level, the pathogenesis of hyperlactatemia varies widely depending on the underlying cause. Microcirculatory dysfunction, mitochondrial dysfunction, and epinephrine-mediated stimulation of Na+ -K+ -ATPase pumps are likely important contributors to hyperlactatemia in critically ill patients. Ultimately, hyperlactatemia is a marker of altered cellular bioenergetics. CONCLUSION: The etiology of hyperlactatemia is complex and multifactorial. Understanding the relevant pathophysiology is helpful when characterizing hyperlactatemia in clinical patients.

Clinical use of plasma lactate concentration. Part 2: Prognostic and diagnostic utility and the clinical management of hyperlactatemia.

OBJECTIVE: To review the current literature pertaining to the use of lactate as a prognostic indicator and therapeutic guide, the utility of measuring lactate concentrations in body fluids other than blood or plasma, and the clinical management of hyperlactatemia in dogs, cats, and horses. DATA SOURCES: Articles were retrieved without date restrictions primarily via PubMed, Scopus, and CAB Abstracts as well as by manual selection. HUMAN AND VETERINARY DATA SYNTHESIS: Increased plasma lactate concentrations are associated with increased morbidity and mortality. In populations with high mortality, hyperlactatemia is moderately predictive in identifying nonsurvivors. Importantly, eulactatemia predicts survival better than hyperlactatemia predicts death. Consecutive lactate measurements and calculated relative measures appear to outperform single measurements. The use of lactate as a therapeutic guide has shown promising results in people but is relatively uninvestigated in veterinary species. Increased lactate concentrations in body fluids other than blood should raise the index of suspicion for septic or malignant processes. Management of hyperlactatemia should target the underlying cause. CONCLUSION: Lactate is a valuable triage and risk stratification tool that can be used to separate patients into higher and lower risk categories. The utility of lactate concentration as a therapeutic target and the measurement of lactate in body fluids shows promise but requires further research.

Retrospective evaluation of cats with elapid snake envenomation associated neurotoxicity requiring mechanical ventilation: 12 cases (2005-2014).

OBJECTIVE: To retrospectively determine the population and outcome characteristics of a cohort of Australian elapid snake envenomed cats requiring mechanical ventilation (MV). DESIGN: Retrospective observational study (2005-2014). SETTING: Academic veterinary emergency and critical care service. ANIMALS: Twelve cats undergoing MV for elapid snake envenomation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The medical records were searched to identify cats requiring MV as part of treatment for elapid snake envenomation. Signalment, the indication for, duration of and complications associated with MV, duration of hospitalization, and survival to hospital discharge were recorded for each of the enrolled cases. Seven cats (58.3%) underwent MV because of presumed unsustainable respiratory effort and 5 cats (41.7%) for respiratory arrest. Eleven cats (91.7%) were successfully weaned from MV and survived to hospital discharge. No cats developed ventilator associated pneumonia or pneumothorax. The median duration of MV was 19.5 hours for the survivors (range 7.0-37.0 hours) and median duration of hospitalization was 3.5 days (range 2.4-14.9 days). CONCLUSIONS: Cats requiring MV for elapid snake envenomation have a favorable outcome and require a relatively short period of MV. Complications encountered are unlikely to influence outcome.

Update: Clinical Use of Plasma Lactate.

Lactate is an essential, versatile metabolic fuel in cellular bioenergetics. In human emergency and critical care, lactate is used as a biomarker and therapeutic endpoint and evidence is growing in veterinary medicine supporting its clinical utility. Lactate production is a protective response providing ongoing cellular energy during tissue hypoperfusion or hypoxia and mitigating acidosis. Hence, hyperlactatemia is closely associated with disease severity but it is an epiphenomenon as the body attempts to protect itself. This article reviews lactate biochemistry, kinetics, pathophysiology, some practical aspects of measuring lactate, as well as its use in diagnosis, prognosis, and monitoring.

Plasma lactate concentration as a prognostic biomarker in dogs with gastric dilation and volvulus.

Initial and serial plasma lactate concentrations can be used to guide decision making in individual dogs with GDV but care is necessary in phrasing conversations with owners. Published data suggests that survival is more likely and the chance of complications less in dogs with an initial plasma lactate of <4 mmol/L. An initial lactate >6 mmol/L makes gastric necrosis and greater expense more likely. However, because of the overlap between groups and the good overall survival rates, exploratory laparotomy should always be recommended irrespective of the plasma lactate concentration. Falls in plasma lactate of greater than ~40% after fluid resuscitation are likely to indicate better survival. If the initial plasma lactate concentration is moderately to severely increased (5->10 mmol/L) and a sustained increase in plasma lactate occurs after fluid resuscitation, the cause should be aggressively pursued. Many dogs with persistent hyperlactatemia over 24-48 hours do not survive.

Effect of body position on the arterial partial pressures of oxygen and carbon dioxide in spontaneously breathing, conscious dogs in an intensive care unit.

OBJECTIVE: To evaluate the effect of body position on the arterial partial pressures of oxygen and carbon dioxide (PaO(2), PaCO(2)), and the efficiency of pulmonary oxygen uptake as estimated by alveolar-arterial oxygen difference (A-a difference). DESIGN: Prospective, randomized, crossover study. SETTING: University teaching hospital, intensive care unit. ANIMALS: Twenty-one spontaneously breathing, conscious, canine patients with arterial catheters placed as part of their management strategy. INTERVENTIONS: Patients were placed randomly into lateral or sternal recumbency. PaO(2) and PaCO(2) were measured after 15 minutes in this position. Patients were then repositioned into the opposite position and after 15 minutes the parameters were remeasured. MEASUREMENTS AND MAIN RESULTS: Results presented as median (interquartile range). PaO(2) was significantly higher (P=0.001) when patients were positioned in sternal, 91.2 mm Hg (86.0-96.1 mm Hg), compared with lateral recumbency, 86.4 mm Hg (73.9-90.9 mm Hg). The median change was 5.4 mm Hg (1.1-17.9 mm Hg). All 7 dogs with a PaO(2)<80 mm Hg in lateral recumbency had improved arterial oxygenation in sternal recumbency, median increase 17.4 mm Hg with a range of 3.8-29.7 mm Hg. PaCO(2) levels when patients were in sternal recumbency, 30.5 mm Hg (27.3-32.7 mm Hg) were not significantly different from those in lateral recumbency, 32.2 mm Hg (28.3-36.0 mm Hg) (P=0.07). The median change was -1.9 mm Hg (-3.6-0.77 mm Hg). A-a differences were significantly lower (P=0.005) when patients were positioned in sternal recumbency, 21.7 mm Hg (17.3-27.7 mm Hg), compared with lateral recumbency, 24.6 mm Hg (20.4-36.3 mm Hg). The median change was -3.1 mm Hg (-14.6-0.9 mm Hg). CONCLUSIONS: PaO(2) was significantly higher when animals were positioned in sternal recumbency compared with lateral recumbency, predominantly due to improved pulmonary oxygen uptake (decreased A-a difference) rather than increased alveolar ventilation (decreased PaCO(2)). Patients with hypoxemia (defined as PaO(2)<80 mm Hg) in lateral recumbency may benefit from being placed in sternal recumbency. Sternal recumbency is recommended to improve oxygenation in hypoxemic patients.

Partial right ventriculectomy using the incised patch technique to treat double chambered right ventricle and chylothorax in a cat.

OBJECTIVE: To report successful surgical treatment of double chambered right ventricle (DCRV) in a cat. STUDY DESIGN: Clinical report. ANIMALS: Cat with DCRV. METHODS: DCRV was diagnosed in a 5-month-old male Maine Coon cat using echocardiography and angiocardiography. At 2 years, despite medical therapy, chylothorax developed. A section of the right ventricular free wall that spanned the fibromuscular obstruction was excised under total venous inflow occlusion (TVIO) using the incised pericardial patch technique. RESULTS: Short-term recovery was complicated by return of chylothorax but this resolved after thoracocentesis and diuretic therapy. Three years after surgery, the cat is free of clinical signs and no longer on any medical therapy. CONCLUSIONS: Partial right ventriculectomy can be performed under TVIO through an incised pericardial patch. CLINICAL RELEVANCE: DCRV in cat can be successfully treated by partial right ventriculectomy performed under TVIO through an incised pericardial patch.

Clinical investigation of a point-of-care blood ammonia analyzer.

BACKGROUND: Hyperammonemia has frequently been implicated in the pathogenesis of hepatic encephalopathy. Blood ammonia determination requires minimal delay between sampling and analysis for accurate results. OBJECTIVES: The aim of this study was to investigate the PocketChem BA, a new point-of-care (POC) blood ammonia analyzer for clinical use by determining machine precision, linearity, repeatability, and accuracy. METHODS: Coefficients of variation were determined by repeated measurement of 2 control solutions. Linearity was investigated by testing serial dilutions of a stock solution. For accuracy, samples from clinical cases were used to compare the results on the PocketChem BA with those obtained using an enzymatic reference method for canine plasma. Canine and feline patients were consecutively enrolled if blood ammonia was assayed and samples could be analyzed shortly after collection. Classification of results (as normal or high, using 100 micromol/L as a cutoff value), Bland-Altman and Deming regression plots, and intraclass correlation coefficients were used to compare the methods. Stability of samples and test strips also was assessed over time. RESULTS: Coefficients of variation were 10.6% and 4.8% for low and high controls, respectively. Concentrations of ammonia in diluted stock solutions correlated positively with mean measured concentrations (Pearson coefficient 0.988, P<.001). Of the 54 samples obtained from 38 dogs and 4 cats, 41 had ammonia concentrations within the readable range. Results from the POC analyzer and the reference method were correlated positively (intraclass coefficient 0.800, 95% confidence interval 0.655-0.888), with the POC analyzer having negative constant and proportional biases. The methods agreed in the classification of 45/54 (83.3%) samples, with 7 false negative results on the POC analyzer. Results of repeated sample and strip analyses at 1 and 24 hours were significantly different (P<.05) from those at 0 hour. CONCLUSIONS: The PocketChem BA has acceptable precision, adequate linearity, and satisfactory agreement with a reference method, but negative constant and proportional biases. The POC analyzer may be suitable for clinical use in patients suspected of having hepatic encephalopathy, using a lower reference limit of 60 mumol/L to decrease false negative results.

Fluid therapy in patients with pulmonary disease.

Fluid therapy in patients with pulmonary disease is challenging. Although a single set of rules cannot be applied to every patient, the following guidelines can be used when managing patients with pulmonary disease. Euvolemic patients with adequate tissue perfusion should be given sufficient isotonic fluid to balance insensible losses. If severe pulmonary compromise is present, cessation of all fluid therapy may be considered if the patient is able to match its losses by voluntary intake. In hypovolemic or hypotensive patients, small boluses of isotonic crystalloids or colloids should be given to restore perfusion, avoiding rates of more than 30 mL/kg an hour for isotonic crystalloids. If perfusion is not restored by adequate volume resuscitation, vasopressors or positive inotropes should be administered to prevent fluid overload and deterioration in pulmonary function.

Acid-base and electrolyte abnormalities in dogs with gastrointestinal foreign bodies.

Gastrointestinal foreign bodies occur commonly in dogs. The objective of the study was to describe the acid-base and electrolyte abnormalities identified in dogs with gastrointestinal foreign bodies and determine if these abnormalities are related to the site or type of foreign body present. Medical records of 138 dogs were reviewed, and information on signalment, initial venous electrolyte and acid-base values, surgical findings, relevant historical information, imaging modalities used, cost of hospital visit, intra- or postoperative complications, and survival was obtained. The site of the foreign body was recorded in 94.9% of cases and the most common site was the stomach (50%), followed by the jejunum (27.5%). The foreign bodies were linear in 36.2% of cases. The most common electrolyte and acid-base abnormalities regardless of the site or type of foreign body were hypochloremia (51.2%), metabolic alkalosis (45.2%), hypokalemia (25%), and hyponatremia (20.5%). No significant association was found between electrolyte or acid-base abnormalities and the site of foreign body. Linear, as opposed to discrete, foreign bodies were more likely to be associated with a low serum sodium concentration (odds ratio, 0.85; 95% confidence interval, 0.75-0.95). Hyperlactatemia (> 2.4 mmol/L) was seen in 40.5% of dogs. A wide variety of electrolyte and acid-base derangements are found in dogs with gastrointestinal foreign bodies. Hypochloremia and metabolic alkalosis are common in these dogs. Hypochloremic, hypokalemic metabolic alkalosis is seen with both proximal and distal gastrointestinal foreign bodies.

Assessment and treatment of perfusion abnormalities in the emergency patient.

Many patients presented to the emergency veterinarian are suffering from global or local tissue hypoperfusion. Global or systemic hypoperfusion can occur secondary to a reduction in the effective circulating intravascular volume (hypovolemic shock) or reduced ability of the heart to pump blood around the body secondary to reduced cardiac function (cardiogenic shock),obstruction to blood flow (obstructive shock), or maldistribution of the circulating intravascular volume (distributive shock). Initial assessment involving physical examination supplemented by measurement of hemodynamic and metabolic parameters allows the clinician to recognize and treat patients with severe global hypoperfusion. Use of techniques like sublingual capnometry and measurement of central venous oxygen saturation may aid recognition and evaluation of early hypoperfusion. Treatment decisions are made based on an assessment of the severity of the hypoperfusion and its probable underlying cause. Early effective treatment of hypoperfusion is likely to lead to a better outcome for the patient.

Veterinary critical care.

Veterinary species experience similar perturbations of their health to those of human patients. When the long-term prognosis is good and providing suffering can be minimized, animals stand to benefit greatly from recent advances in the field of emergency and critical care. Outcomes in many conditions in small and large animals have improved markedly in the last 15 years, as management has improved, making the financial and emotional investment in critical care worthwhile for many owners.