Changes in Hemostatic Indices in Foals Naturally Infected With Strongylus vulgaris

Strongylus vulgaris has been found endemic in equine populations subject to parasite control by targeted selective anthelmintic therapy. This study investigated hemostasis in foals naturally infected with S. vulgaris and monitored this response over the course of progressing infection stages. The hemostatic indices D-dimer, antithrombin III (ATIII), fibrinogen, prothrombin time (PT), and activated partial thromboplastin time were evaluated in weekly blood samples for up to 50 weeks in 12 foals born into a herd with high prevalence of S. vulgaris. Results were compared with weekly S. vulgaris antibody enzyme-linked immunosorbent assay values in all foals using a linear mixed effects model with repeated measures and to total numbers of S. vulgaris larvae in nine foals at necropsy with Pearson linear correlation. In the first week of life, all evaluated indices of hemostasis were significantly different from those observed in the rest of the study weeks, corresponding to previously demonstrated aberrancies in neonates. Significant changes were seen for D-dimer in weeks 11–24, 26–27, 30, and 39 compared with week 2, for PT in weeks 12–13 compared with week 6, and for ATIII in week 15 compared with week 4. Strongylus vulgaris antibody levels were statistically associated with D-dimer (P = .0076) and fibrinogen (P = .0004) concentrations. Naturally acquired infection with S. vulgaris was associated with changes suggestive of mild activation of coagulation, fibrinolysis, and inflammation. The results of this study may help elucidate the pathogenesis of the endarteritis, thromboembolism, and nonstrangulating intestinal ischemia that is observed in horses with S. vulgaris infection.     

Calcium Regulating Hormones and Serum Calcium and Magnesium Concentrations in Septic and Critically Ill Foals and their Association with Survival

Background: Disorders of calcium regulation are frequently found in humans with critical illness, yet limited information exists in foals with similar conditions including septicemia. The purpose of this study was to determine whether disorders of calcium exist in septic foals, and to determine any association with survival.
Hypothesis: Blood concentrations of ionized calcium (Ca²⁺) and magnesium (Mg²⁺) will be lower in septic foals with concomitant increases in parathyroid hormone (PTH), calcitonin (CT), and parathyroid-related peptide (PTHrP) compared with healthy foals. The magnitude of these differences will be negatively associated with survival.
Animals: Eighty-two septic, 40 sick nonseptic, and 24 healthy foals of ≤7 days were included.
Methods: Prospective, observational study. Blood was collected at initial examination for analysis. Foals with positive blood culture or sepsis score ≥14 were considered septic. Foals with disease other than sepsis and healthy foals were used as controls. Hormone concentrations were measured with validated immunoassays.
Results: Septic foals had decreased Ca²⁺ (5.6 versus 6.1 mg/dL, P < .01) and increased serum PTH (16.2 versus 3.2 pmol/L, P < .05), and phosphorus concentrations (7.1 versus 6.3 mg/dL, P < .01). No differences in serum Mg²⁺, PTHrP, and CT concentrations were found. Nonsurviving septic foals (n = 42/82) had higher PTH concentrations (41.1 versus 10.7 pmol/L, P < .01) than survivors (n = 40/82).
Conclusions and Clinical Importance: Septic foals were more likely to have disorders of calcium regulation compared with healthy foals, where hyperparathyroidemia was associated with nonsurvival.     

Uveal Inflammation in Septic Newborn Foals

Background: Septicemia in humans is described as a leading cause of uveitis, which eventually can induce blindness. Hypothesis/Objectives: Uveal inflammatory findings could be related to sepsis severity in newborn foals and might be used as an indirect indicator for survival. Animals: Seventy‐four septic foals, 54 nonseptic foals, and 42 healthy foals. Methods: Prospective observational clinical study. A detailed blinded, ophthalmic examination was performed by boarded ophthalmologists on all admitted newborn foals. Foals were grouped as septic (when blood culture resulted positive or the sepsis score was ≥14), nonseptic, and controls. Based on blood culture results, the septic group was subdivided into bacteremic and nonbacteremic foals. Results: Blood culture was performed in 62/74 septic foals, from which 35 (56%) were bacteremic and 27 (44%) were nonbacteremic. Anterior uveitis was diagnosed in a significantly (P < .005) higher number of septic/bacteremic foals (14/35, 40%) than in septic/nonbacteremic foals (5/27, 19%), nonseptic foals (4/54, 7%), and control foals (0%). Anterior chamber fibrin was only observed in 4/14 (29%) septic/bacteremic foals with anterior uveitis. Anterior uveitis was also associated with posterior uveitis in 6/35 (19%) septic/bacteremic foals. The diagnosis of uveitis was related to nonsurvival (P= .001, odds ratio = 6.2, 95% confidence interval = 2.1–18.2). Conclusions and Clinical Importance: Anterior uveitis is highly prevalent in septic newborn foals, especially in those with a positive blood culture, and it should be considered as a survival prognostic factor.     

Sick Neonatal Foals Do Not Demonstrate Evidence of Oxidative Stress

Critical illness in humans is associated with alterations in oxidative stress and the concentration of antioxidant molecules; however, this association has not been examined in equine neonates. The purpose was to determine the concentration of various antioxidant molecules, as well as a marker of oxidative stress, in the serum and plasma of normal and sick neonatal foals and their dams. Results demonstrated that the concentration of selenium was less (61.71 vs. 77.93 ng/mL; P = .002) in sick versus healthy neonates, whereas the concentration of vitamin E was slightly higher in sick compared with healthy foals (4.36 vs. 3.17 mg/mL); however, this did not achieve statistical significance (P = .31). The vitamin E concentration was greater (5.37 vs. 3.43 mg/mL; P = .01) and serum selenium was less in sick mares (129.50 vs. 184.78 ng/mL; P = .0001) compared with healthy mares. In addition, the serum concentration of selenium is lower in neonates than in their dams in the perinatal period (70.10 vs. 173.34 ng/mL; P = .0001). Glutathione peroxidase activity was less in sick foals (7.04 nmol/min/mL) compared with healthy foals (9.13 nmol/min/mL) (P = .19), and 3-nitrotyrosine (3-NT) concentration/mg protein was less in sick foals versus healthy foals (geometric mean, 1.24 vs. 2.07 nmol 3-NT/mg protein). This difference did not achieve statistical significance (P = .09); however, when a subset of critically ill foals was examined, the assayed concentration of 3-NT/mg protein was even less (0.99 nmol 3-NT/mg protein) and did statistically differ from healthy foals (P = .03).     

Blood arginine vasopressin, adrenocorticotropin hormone, and cortisol concentrations at admission in septic and critically ill foals and their association with survival

BACKGROUND: Sepsis is an important cause for neonatal foal mortality. The hypothalamic-pituitary-adrenal axis (HPAA) responses to sepsis are well documented in critically ill humans, but limited data exist in foals. The purpose of this study was to evaluate the HPAA response to sepsis in foals, and to associate these endocrine changes with survival. HYPOTHESIS: Blood concentrations of arginine vasopressin (AVP), adrenocorticotropin hormone (ACTH), and cortisol will be higher in septic foals as compared with sick nonseptic and healthy foals. The magnitude of increase in hormone concentration will be negatively associated with survival. ANIMALS: Fifty-one septic, 29 sick nonseptic, and 31 healthy foals of < or =7 days of age were included. METHODS: Blood was collected at admission for analysis. Foals with positive blood culture or sepsis score > or =14 were considered septic. Foals admitted with disease other than sepsis and healthy foals were used as controls. AVP, ACTH, and cortisol concentrations were measured using validated immunoassays. RESULTS: AVP, ACTH, and cortisol concentrations were increased in septic foals. Septic nonsurvivor foals (n = 26/51) had higher plasma ACTH and AVP concentrations than did survivors (n = 25/51). Some septic foals had normal or low cortisol concentrations despite increased ACTH, suggesting relative adrenal insufficiency. AVP, ACTH, and cortisol concentrations were higher in sick nonseptic foals compared with healthy foals. CONCLUSIONS AND CLINICAL IMPORTANCE: Increased plasma AVP and ACTH concentrations in septic foals were associated with mortality. Several septic foals had increased AVP : ACTH and ACTH : cortisol ratios, which indicates relative adenohypophyseal and adrenal insufficiency.     

Bacterial Endocarditis in Two Spanish Foals After Neonatal Septicemia

Equine neonatal septicemia could lead to the release of thrombus and heart valvular endocarditis. A relationship between activation of the renin-angiotensin-aldosterone-vasopressin (RAAV) axis and heart failure has been described in several species. This article describes the echocardiographic, electrocardiographic, and laboratory findings, including RAAV axis, in two Spanish foals with endocarditis after septicemia in comparison with two control groups matched by age and gender. Two Spanish foals (F1, 60-day-old colt; F2, 76-day-old filly) were presented with poor growth rate a month after being hospitalized because of septicemia. Colt F1 had hypercortisolemia and increased left ventricular internal dimensions in systole and diastole, increased left ventricular free wall thickness in diastole, interventricular septum thickness in diastole, and mitral valve thickness. Colt F1 also presented lower fractional shortening, ejection fraction, fractional wall thickness, and fractional septum thickness. Filly F2 showed hyperfibrinogenemia and increased values for aspartate aminotransferase, lactate dehydrogenase, creatin kinase, alkaline phosphatase, interventricular septum at systole, aorta diameter at the level of valve leaflets and sinotubular junction, and mitral and tricuspid valve thickness. The foals did not present systemic signs compatible with heart failure, heart arrhythmias were not found, and RAAV values were within the expected confidence limits. In conclusion, endocarditis without clinical signs of heart failure can appear shortly after septicemia in equine neonates. The measurement of the thickness of the valves by two-dimensional echocardiography is diagnostic. The lack of activation of RAAV in both foals might indicate that the severity of the heart problem was mild.     

Plasma Adrenomedullin Concentrations in Critically Ill Neonatal Foals

Background

Bacterial sepsis remains a leading cause of morbidity and mortality in neonatal foals, but accurate diagnostic and prognostic markers are lacking. Adrenomedullin (AM) is a polypeptide with diverse biologic effects on the cardiovascular system that increases in septic humans and laboratory animals.

Hypotheses

Plasma AM concentration (p[AM]) is increased in septic neonatal foals compared to sick nonseptic and healthy control foals, and p[AM] is predictive of survival in septic neonatal foals.

Animals

Ninety critically ill (42 septic, 48 sick nonseptic) and 61 healthy foals <1 week of age.

Methods

A prospective observational clinical study was performed. Venous blood was collected from critically ill foals at admission and from healthy foals at 24 hours of age. Critically ill foals were categorized as septic or sick nonseptic based on blood culture results and sepsis score. Plasma [AM] was measured by using a commercially available ELISA for horses. Data were analyzed by using the Mann‐Whitney U‐test and P < .05 was considered significant.

Results

Plasma [AM] was not significantly different between septic and sick nonseptic foals (P = .71), but critically ill foals had significantly increased p[AM] compared to healthy controls (P < .0001). In critically ill foals, p[AM] was not predictive of survival (P = .051). A p[AM] cutoff concentration of 0.041 ng/mL provided a test sensitivity of 91% and specificity of 54% to predict illness.

Conclusions and Clinical Relevance

Plasma [AM] shows promise as a marker of health in neonatal foals, but p[AM] increases nonspecifically during perinatal illnesses and is not necessarily associated with sepsis.     

Prospective Evaluation of Coagulation in Critically Ill Neonatal Foals

Background: Coagulopathy is a potentially underrecognized complication of sepsis and septic shock in critically ill neonatal foals.
Hypothesis: Critically ill neonatal foals have abnormalities in coagulation that are associated with disease severity and outcome.
Animals: Foals <72 hours old admitted to a neonatal intensive care unit.
Methods: Prospective, observational study. Blood was collected at admission, 24, and 48 hours for platelet count, prothrombin time, activated partial thromboplastin time, antithrombin activity and concentrations of fibrin degradation products, and fibrinogen in plasma from all foals.
Results: Sixty-three foals were enrolled and classified as Septic Shock (12), Septic (28), and Other (23). At least 1 abnormal value was found in 18/28 (64%) samples from the Septic Shock group, 66/85 (78%) from the Septic group, and 30/59 (51%) from the Other group ( P = .01). Coagulopathy (3 or more abnormal values) was present in 7/28 (25%) samples in the Septic Shock group, 14/85 (16%) samples in the Septic group, and 3/59 (5%) samples in the Other group ( P = .0028). Clinically detectable bleeding occurred in 8/12 (67%) Septic Shock cases, 11/28 (39%) Septic cases, and 3/23 (13%) Other cases ( P = .009). Foals in Septic Shock were 12.7 times more likely to have clinical evidence of bleeding than those in the Other group (95% CI 2.3–70, P = .004). Treatment with fluids or plasma did not have a detectable effect on coagulation values.
Conclusions and Clinical Importance: Coagulopathy commonly occurs in critically ill neonatal foals, especially those with sepsis and septic shock.     

Hypothalamic-Pituitary-Adrenal Axis Dysfunction in Hospitalized Neonatal Foals

Background: Transient hypothalamic-pituitary-adrenal (HPA) axis dysfunction occurs frequently in critically ill humans and impacts survival. The prevalence and impact of HPA axis dysfunction in critically ill neonatal foals are not well characterized.
Hypotheses: (1) HPA axis dysfunction occurs in hospitalized neonatal foals, and is characterized by inappropriately low basal serum cortisol concentration or inadequate cortisol response to exogenous adrenocorticotropic hormone (ACTH); (2) hospitalized foals with HPA axis dysfunction have more severe disease and are less likely to survive than hospitalized foals with normal HPA axis function.
Animals: Seventy-two hospitalized foals and 23 healthy age-matched foals.
Methods: Basal ACTH and cortisol concentrations were measured and a paired low-dose (10 μg)/high-dose (100 μg) cosyntropin stimulation test was performed at admission in hospitalized foals. HPA axis dysfunction was defined as (1) an inappropriately low basal cortisol concentration or (2) an inadequate increase in cortisol concentration (delta cortisol) after administration of cosyntropin, with cut-off values for appropriate basal and delta cortisol concentrations determined from results obtained in healthy age-matched foals.
Results: Forty-six percent of hospitalized foals had an inappropriately low basal cortisol concentration and 52% had an inadequate delta cortisol concentration after administration of the 100 μg dose of cosyntropin. An inadequate delta cortisol response to the high (100 μg) dose of cosyntropin was significantly correlated with shock and multiple organ dysfunction syndrome in hospitalized foals, and with decreased survival in a subgroup of septic foals.
Conclusions and Clinical Importance: HPA axis dysfunction occurs frequently in hospitalized neonatal foals, and negatively impacts disease severity and survival.     

Vaccination Response of Young Foals to Keyhole Limpet Hemocyanin: Evidence of Effective Priming in the Presence of Maternal Antibodies

The ability to produce antibodies is essential for protection from infectious disease; however, in the neonate, maternal antibodies have been proposed to interfere with the foal's ability to respond to vaccination. In species other than the equid, keyhole limpet hemocyanin, a high-molecular weight protein, is used in vivo as an experimental vaccine component because of its high intrinsic immunogenicity. In this study, we show that young foals are able to produce a primary antibody response to vaccination at an early age. Thus, foals, like human infants, are capable of responding to antigenic exposure to a novel antigen (keyhole limpet hemocyanin) during the neonatal period. Although vaccinating foals in the presence of maternal antibodies failed to induce a primary serological response, priming occurred as comparable anamnestic responses were detected upon subsequent exposure to the antigen. There was no evidence of tolerance induction.     

Efficacy and Safety of a Commercial Fresh-Frozen Hyperimmune Plasma in Foals With Failure of Passive Transfer of Immunity

In foals more than 12 hours old, the only effective therapy for the treatment of failure of passive transfer (FPT) of immunity is transfusion of equine plasma. Use and efficacy of equine plasma for prophylaxis and treatment of sepsis, a condition primarily associated with FPT, are widely reported. However, plasma- and recipient-related factors associated with extent of IgG transfer and catabolism are not completely defined. Efficacy and safety of transfusion of a commercial fresh-frozen hyperimmune plasma were evaluated in hospitalized foals younger than 7 days of age with total or partial FPT. Sixty-two foals, classified as affected by FPT only, septic (infection plus systemic inflammatory response syndrome [SIRS]), and nonseptic sick, were included, and serum IgG concentration was measured at admission and 24 hours after plasma transfusion. In 25/62 foals, IgG level after 72 hours was also determined. The impact of different classification criteria for septic foals on IgG transfer was evaluated. Serum IgG measured 24 hours and 72 hours after plasma transfusion was significantly greater than at admission, but no significant difference was found in transfer efficacy (TE) between FPT, FPT septic, and FPT nonseptic foals and no significant difference was found in IgG concentration comparing foals with total and partial FPT or survivors and nonsurvivors. No significant difference was found comparing IgG concentration between bacteremic and nonbacteremic foals and foals with or without SIRS. No foal experienced adverse reactions to plasma transfusion. IgG TE and catabolism did not result significantly affected by the presence of sepsis or illness or by the outcome.     

Blood and Cerebrospinal Fluid α‐Tocopherol and Selenium Concentrations in Neonatal Foals with Neuroaxonal Dystrophy

Background

Equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (NAD/EDM) is a neurodegenerative disorder affecting genetically predisposed foals maintained on α‐tocopherol (α‐TP)‐deficient diet.

Objective

Intramuscular α‐TP and selenium (Se) administration at 4 days of age would have no significant effect on serum or cerebrospinal fluid (CSF) α‐TP in healthy foals. Serum and CSF α‐TP, but not Se, would be significantly decreased in NAD/EDM‐affected foals during first year of life.

Animals

Fourteen Quarter horse foals; 10 healthy foals supplemented with 0.02 mL/kg injectable α‐TP and Se (n = 5) or saline (n = 5) at 4 days of age and 4 unsupplemented NAD/EDM‐affected foals.

Methods

Complete neurologic examinations were performed, blood and CSF were collected before (4 days of age) and after supplementation at 10, 30, 60, 120, 180, 240, and 360 days of age. Additional blood collections occurred at 90, 150, 210, and 300 days. At 540 days, NAD/EDM‐affected foals and 1 unsupplemented healthy foal were euthanized and necropsies performed.

Results

Significant decreases in blood, CSF α‐TP and Se found in the first year of life in all foals, with most significant changes in serum α‐TP from 4–150 days. Dam α‐TP and Se significantly influenced blood concentrations in foals. Injection of α‐TP and Se did not significantly increase CSF Se, blood or CSF α‐TP in healthy foals. NAD/EDM‐affected foals had significantly lower CSF α‐TP through 120 days.

Conclusions and Clinical Importance

Injection of α‐TP and Se at 4 days of age does not significantly increase blood or CSF α‐TP. Despite all 14 foals remaining deficient in α‐TP, only the 4 genetically predisposed foals developed NAD/EDM.     

Plasma C‐Reactive Protein and Haptoglobin Concentrations in Critically Ill Neonatal Foals

Background

Accurate diagnostic markers for sepsis in neonatal foals are needed. Plasma C‐reactive protein concentration (p[CRP]) and haptoglobin concentration (p[Hp]) are well‐established biomarkers of infection in humans, but studies are lacking in foals.

Hypotheses

p[CRP]) and p[Hp] are increased in septic foals compared to sick nonseptic and healthy control foals, and are predictive of survival.

Animals

Eighty critically ill foals (40 septic, 40 sick nonseptic) and 39 healthy control foals <1 week of age.

Methods

Multicenter, prospective observational clinical study. Venous blood was collected at admission from septic and sick nonseptic foals and from clinically healthy foals at 24 h of age. A diagnosis of sepsis was made based on positive blood culture or a sepsis score >11, and p[CRP] and p[Hp] were measured by using ELISA tests. Data were analyzed by using the Mann‐Whitney U‐test and forward stepwise multivariable linear regression. P < .05 was considered significant.

Results

Plasma [CRP] was positively associated with age, serum globulin, adrenomedullin, and bilirubin concentrations, aspartate aminotransferase activity, glutamyl‐transferase activity, band neutrophil count, and rectal temperature, and was increased in foals with toxic neutrophils, enterocolitis, colic, rib fractures and septic arthritis. Surprisingly, p[Hp] was lower in septic foals than in sick nonseptic foals. Neither p[CRP] or p[Hp] was predictive of survival in critically ill foals.

Conclusions and Clinical Importance

Plasma [CRP] increases with inflammation in neonatal foals but is not indicative of sepsis. Single time point, admission sampling of p[CRP] and p[Hp] do not appear to be useful biomarkers for sepsis in foals.     

Peripheral Blood Lymphocyte Subpopulations and Immunoglobulin Concentrations in Healthy Foals and Foals with Rhodococcus equi Pneumonia

Infectious diseases are common in foals aged 1-5 months. The objectives of this investigation were to evaluate immunologic parameters in foals from birth to weaning to establish reference values for the proportion of circulating lymphocytes that were helper (CD4+) or cytotoxic (CD8+) T cells, or B cells; to measure serum immunoglobulin (IgM and IgG) concentrations; and to compare these immunologic parameters to values in foals with naturally occurring Rhodococcus equi pneumonia and in adult horses. Peripheral blood lymphocyte subpopulations were determined by flow cytometric analysis, and serum IgG and IgM concentrations were determined by radial immunodiffusion. Flow cytometric analysis of lymphocyte subpopulations suggested age-related changes in the cell-mediated immune system in horses. Absolute circulating CD4+ and CD8+ T lymphocytes and B cells increased linearly up to 3 months of age. Circulating B cell concentrations from birth to 6 months of age were greater than values in adult horses and the lymphocyte differences among the age groups are mainly due to variation in B lymphocytes. Both absolute and proportional B cell concentrations were greater in foals with R equi pneumonia than in healthy foals at the same age. The increase in absolute cell counts of each subpopulation was dependent on the increase of absolute peripheral blood lymphocyte count. Serum IgG concentration increased linearly from 1 to 3 months of age, and serum IgM concentrations increased from 1 to 6 months of age. These data suggest age-dependent cell-mediated and humoral development in young foals.