Influence of age on neutrophil function in foals.

Functional activities (phagocytosis and killing) of neutrophil leucocytes (NL) and immunoglobulin G concentrations were evaluated in six healthy foals from birth to 6 months of age. Peripheral blood NL were reacted with Streptococcus equisimilis in 20 per cent pooled equine serum for 30, 60 and 90 mins and functional activities of NL were determined using a fluorochrome microassay. Values for foal NL function were compared with those of healthy adult horses (n = 28). Foal neutrophil function was influenced by age. Killing capacity of NL decreased, whereas phagocytic capacity increased, until 113 days of age, after which a reversal in trends became apparent. Immunoglobulin G concentrations changed significantly over time and were lowest at 29 to 56 days of age. All foal values for NL function fell within the range of normal values established for healthy adult horses.     

Serum free cortisol fraction in healthy and septic neonatal foals

Background: Relative cortisol insufficiency occurs in septic foals and impacts survival. Serum free (biologically available) cortisol concentration might be a better indicator of physiologic cortisol status than serum total cortisol concentration in foals. Hypotheses: In septic foals, (1) low free cortisol concentration correlates with disease severity and survival and (2) predicts disease severity and outcome better than total cortisol concentration. Animals: Fifty‐one septic foals; 11 healthy foals; 6 healthy horses. Methods: In this prospective clinical study, foals meeting criteria for sepsis at admission were enrolled. University‐owned animals served as healthy controls. Basal and cosyntropin‐stimulated total cortisol concentration and percent free cortisol (% free cortisol) were determined by chemiluminescent immunoassay and ultrafiltration/ligand‐binding methods, respectively. Group data were compared by ANOVA, Mann‐Whitney U‐tests, and receiver operator characteristic curves. Significance was set at P < .05. Results: Basal % free cortisol was highest in healthy foals at birth (58±8% mean±SD), and was higher (P≤.004) in healthy foals of all ages (33±6 to 58±8%) than in adult horses (7±3%). Cosyntropin‐stimulated total and free cortisol concentrations were lower (P≤.03) in foals with shock (total = 6.2±8.1 μg/dL; free = 3.5±4.8 μg/dL versus total = 10.8±6.0 μg/dL; free = 6.9±3.3 μg/dL in foals without shock) and in nonsurvivors (total = 3.8±6.9 μg/dL; free = 1.9±3.9 μg/dL versus total = 9.1±7.7 μg/dL; free = 5.5±4.4 μg/dL in survivors). Free cortisol was no better than total cortisol at predicting disease severity or outcome in septic foals. Conclusions and Clinical Importance: Serum free cortisol is impacted by age and illness in the horse. There is no advantage to measuring free over total cortisol in septic foals.     

Thromboelastography in healthy, sick non-septic and septic neonatal foals

Objectives To evaluate citrated recalcified thromboelastography (TEG) in healthy newborn foals, and to determine intra‐assay, inter‐individual and intra‐individual (at 12 h, 24 h and 7 days after birth) variations. Additionally, to compare TEG variables, haematological values and conventional coagulation profiles from healthy, sick non‐septic, and septic foals. Design Prospective study. Methods The study group comprised 18 healthy, 15 sick non‐septic and 17 septic foals. Two citrated (3.2%; 1 : 9 anticoagulant : blood ratio) blood samples were submitted for haemostatic evaluation using a TEG analyser and conventional coagulation profile. TEG values (R time (R), K time (K), angle (α), maximum amplitude (MA) and G value (G)), complete blood count (CBC) and conventional coagulation profile (prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen concentration (Fib) and antithrombin (AT)) were evaluated. Signalment, presenting complaint, sepsis scores, blood culture results and outcome were taken from the medical records of the sick foals. Results Mean values ± SD for TEG variables in healthy neonatal foals were: R = 11.82 ± 5.35 min, K = 3.06 ± 1.34 min, α= 51.19 ± 12.66 degrees, MA = 55.06 ± 6.67 mm and G = 6361 ± 1700 dyn/cm². Mean coefficients of variation for intra‐assay/inter‐individual/intra‐individual in healthy foals were: R = 3.5/45.2/43.1%; K = 5.3/58.7/28.7%; α= 1.5/24.7/11.9%; MA = 0.3/12.1/6.1%; G = 1.6/26.7/14.7%. Septic foals had significantly greater α, MA and G values than sick non‐septic foals, and significantly greater MA and G than healthy foals, changes that are consistent with hypercoagulability. Weak correlations were detected between TEG variables and haematological or haemostatic values. Conclusions TEG could be used to provide additional information about the haemostatic system in equine neonates.     

Evaluation of granulocyte transfusion in healthy neonatal pony foals

Granulocyte transfusions (GT), 0.98 X 10(9) neutrophils/kg of body weight, were performed on 7 healthy pony foals between 2 and 7 days old. The mean neutrophil count of the foals was significantly (P less than 0.05) greater than base line (4,830 +/- 1,260/microliter) 1 hour after GT (8,870 +/- 3,350/microliter) and was similar to base line by 15 to 18 hours after GT (6,550 +/- 2,310/microliter). Leukocyte concentrates (LC) used for GT were harvested from clinically normal adult horses by continuous-flow centrifugation leukapheresis (CL), 3 to 6 hours after hydrocortisone sodium succinate was administered to increase the blood neutrophil count. The mean neutrophil count of the LC used for GT was 68,050 +/- 13,990/microliter, and the mean LC volume was 377.4 +/- 79.2 ml (14.82 +/- 3.54 ml/kg). The mean time required to collect the LC used for GT was 232.1 +/- 73.4 minutes. Neutrophils from LC had significantly reduced in vitro stimulated migration to zymosan-activated serum, when compared with peripheral blood neutrophils of the donors (P less than 0.05). Neutrophil phagocytosis and bactericidal capacity were not significantly changed in LC. Mean neutrophil migration indices were not significantly different in foals after GT. Mild depression and transient diarrhea was noticed in 1 foal 30 minutes after the start of the GT. The donor of LC for this foal and 1 other donor experienced depression, piloerection, and muscle tremors during CL, indicating that complement had been activated. Problems were eliminated by the use of new disposable plastic materials for blood processing in each CL procedure.     

Effect of age on the concentrations of amino acids in the plasma of healthy foals

The concentrations of 23 amino acids in the plasma of 13 healthy foals were determined before suckling, when foals were 1 to 2 days old, 5 to 7 days old, 12 to 14 days old, and 26 to 28 days old. The ratio of the branched chain amino acids to the aromatic amino acids was also calculated at the 5 time points. Analysis of the concentrations at the 5 ages revealed a significant temporal relationship for each amino acid ranging from a polynomial order of 1 to 4 inclusively. There were significant differences between several concentrations of amino acids in plasma at specific sample times; however, no consistent patterns were revealed. The concentrations of amino acids in healthy foals were markedly different from previously determined values in adult horses. The significant differences in the concentrations of amino acids in plasma of healthy foals at the 5 ages may represent developmental aspects of amino acid metabolism or nutrition.     

Influence of age and plasma treatment on neutrophil phagocytosis and CD18 expression in foals

The aim of this study was to evaluate the influence of age and plasma treatment on neutrophil phagocytosis, CD18 expression and serum opsonic capacity in foals in field settings. Microbial infections constitute a large threat in young foals and neutrophil functions are crucial for the defense. Blood samples were obtained from 13 foals at seven time points between the ages of 2 and 56 days and once from 16 adult horses. Six of the foals were treated with adult plasma at the age of 1 week. Neutrophil phagocytosis of yeast after various opsonizations and the expression of complement adhesion receptor CD18 were analysed by flow cytometry. Autologous serum opsonization resulted in 52+/-6.1% phagocytic neutrophils in 2-day-old foals (n = 12), a significantly lower rate than in adult horses (mean 84+/-3.1%; n = 16). In foals, yeast ingestion per neutrophil was also lower than in adults. Opsonic capacity increased with age (p < 0.05), reaching adult levels at 3-4 weeks. An increase in serum opsonic capacity followed plasma treatment (p < 0.05). The phagocytic capacity of foal neutrophils at the time-points studied was equal to or higher than that in the adults, when pooled adult horse serum or anti-yeast IgG was used as opsonin. In foals, serum IgG concentration was negatively correlated to serum opsonic capacity. CD18 receptor expression was higher in neutrophils from foals (<21 days old) than in those from adult horses (p < 0.05). The results indicate that foals are transiently deficient in serum opsonic capacity, which negatively affects their capacity for neutrophil phagocytosis. These changes in serum opsonins, unrelated to IgG, may be important factors in susceptibility to infections in foals.     

Characterization of peripheral blood and pulmonary leukocyte function in healthy foals

Studies in infants and foals indicate an age-dependent maturation of peripheral lymphocyte subsets. The age-dependent relationship for maturation of cellular immune responses, such as phagocytosis and lymphocyte responses of the peripheral and pulmonary-derived leukocytes, has not been characterized in foals. Lymphocyte subpopulations, mitogen stimulation response of lymphocytes, lymphokine-activated killing cell activity, phagocytosis and oxidative burst activity, and serum immunoglobulin (Ig) classes G and M concentrations were determined in developing foals. This study illustrates age-dependent changes in immunoglobulin class concentrations, lymphocyte subsets, and EqMHC Class II expression in cells of the peripheral blood and lungs of developing neonatal-to-weanling foals. The increase in peripheral blood and BAL B-lymphocytes and serum immunoglobulins in developing foals suggests expansion of immune cell populations during a time in which environmental pathogen exposure is great. General immune function, mitogenic responses, LAK cell activity, opsonized phagocytosis, and oxidative burst activity of newborns was similar to the adult horse. Total immune-cell numbers, rather than function, seemed to be the limiting factor in the development of the equine neonatal immune system. There was an age-related percent increase in the appearance of pulmonary lymphocytes, but a percent decrease in macrophages. Although development of the respiratory immune system follows changes in the peripheral blood, cellular expansion, activation, and migration may occur at a slower pace, making the respiratory environment susceptible to pathogens prior to optimal immune system maturity.     

The effect on creatinine kinase activity of freezing plasma from healthy foals

AIM: To assess the stability of creatine kinase (CK) activity in plasma collected from healthy foals and frozen at -20 degrees C for up to 12 weeks. METHODS: Samples of venous blood drawn from 25 foals were analysed for CK activity soon after collection, and again after 1 and 12 weeks of freezing at -20 degrees C. RESULTS: CK activity decreased (p<0.001) between Week 0 and Week 1 and between Week 0 and Week 12. CONCLUSIONS AND CLINICAL RELEVANCE: Decreases in CK activity were statistically significant but clinically insignificant.     

The effect on creatinine kinase activity of freezing plasma from healthy foals

Abstract

AIM: To assess the stability of creatine kinase (CK) activity in plasma collected from healthy foals and frozen at ?20°C for up to 12 weeks.

METHODS: Samples of venous blood drawn from 25 foals were analysed for CK activity soon after collection, and again after 1 and 12 weeks of freezing at ?20°C.

RESULTS: CK activity decreased (p<0.001) between Week 0 and Week 1 and between Week 0 and Week 12.

CONCLUSIONS AND CLINICAL RELEVANCE: Decreases in CK activity were statistically significant but clinically insignificant.     

Glucose metabolism in five septic neonatal foals

Objective: Glucose metabolism is often deranged in septic animals. Bacteremia and sepsis are common in foals and clinical experience suggests that glucose metabolism is abnormal in some of these animals. The purpose of this study was to provide initial estimates of rates of glucose appearance, disappearance, and metabolic clearance rate in septic foals.
Series Summary: Rates of glucose entry, and exit from blood were determined by use of infusion of isotopically labeled glucose in 5 foals with confirmed sepsis. Serum concentrations of glucose, insulin, glucagon, and cortisol were measured concurrent with measurement of rates of glucose turnover. Median glucose turnover rate was 24 μmol/kg/min (range 17–53 μmol/kg/min), and median glucose metabolic clearance rate was 3.2 mL/kg/min (range 1.7–6.7 mL/kg/min). Median concentration of serum immunoreactive insulin was 55 pmol/L (range 36–190 pmol/L), median serum immunoreactive glucagon was 65 pmol/L (range 19–120 pmol/L), and median serum cortisol was 207 nmol/L (range 100–333 nmol/L).
New or unique information provided: These data, although limited in scope and by the lack of data in healthy foals, demonstrate the magnitude and variation in glucose appearance, disappearance, and metabolic clearance rate in septic foals, provide an estimate of rates of glucose utilization in sick foals, and will be useful in guiding future studies of energy metabolism in healthy and ill foals.     

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.     

Debridement of septic physeal lesions in 3 foals

Radiographically, osteolysis of the physis consistent with a septic physitis was observed in 3 foals. The foals were treated with surgical debridement and antimicrobials. Two of the horses were sound for use as adults; the third was euthanatized due to concurrent infectious arthritis and septicemia.     

In vivo effect of ascorbic acid on neutrophil function in healthy and dexamethasone-treated cattle

Ascorbic acid (20 mg/kg of body weight) administered subcutaneously to otherwise nontreated cattle resulted in enhancement of neutrophil oxidative metabolism and capability of neutrophils to mediate antibody-dependent cell-mediated cytotoxicity (ADCC). Random migration, bacterial ingestion, and iodination by neutrophils was unaffected. Three dosage levels of ascorbic acid (10 mg/kg, 20 mg/kg, and 40 mg/kg) were examined for their effects on neutrophil function in cattle treated with dexamethasone (0.04 mg/kg). Dexamethasone administration caused an enhancement of neutrophil random migration and a suppression of neutrophil oxidative metabolism, iodination, and ADCC. None of the dosage levels of ascorbic acid had an effect on the alterations in the WBC count induced by dexamethasone. The ascorbic acid did tend to reverse the effects of dexamethasone on neutrophil random migration, oxidative metabolism, and ADCC in a dose-dependent manner, with the lowest dose having no discernible effect. Ascorbic acid administration also tended to enhance Staphylococcus aureus ingestion by bovine neutrophils. These results indicate that ascorbic acid should be further investigated for its potential to reduce the susceptibility of stressed or glucocorticoid-treated cattle to infective processes.     

Effect of Cysticercus cellulosae on neutrophil function and death

Neutrophils, eosinophils and macrophages interact with invading parasites and naive hosts. The initial reaction of leukocytes is the generation of reactive oxygen species (ROS). The cytotoxic effects of extracts derived from intact Cysticercus cellulosae and from the scolex or membrane fractions on neutrophils were examined. DNA fragmentation of neutrophils was observed when cells were incubated with an extract from the intact metacestode; however, the addition of antioxidant enzymes to the incubation medium had a protective effect. The scolex and membrane extracts did not affect DNA fragmentation of neutrophils. Hydrogen peroxide production of neutrophils incubated with metacestode fractions from C. cellulosae increased by 190% (total extract), 120% (scolex) or 44% (membrane). An increase in antioxidant catalase activity (28%) concomitant with the increased production of ROS was observed in neutrophils incubated with metacestode fractions, which could be an attempt at self-protection. ROS production by neutrophils in the presence of the intact cysticerci extract did not alter phagocytosis. In contrast, the scolex and membrane fractions increased the phagocytic capacity of neutrophils by 44 and 28%, respectively. The results showed that the extract from intact C. cellulosae was toxic for neutrophils via ROS production, leading to DNA fragmentation and inhibition of phagocytic capacity, but neutrophils are able to protect themselves against oxidative stress by via catalase activity.