Serum amyloid A (SAA) as an aid in the management of infectious disease in the foal: comparison with total leucocyte count, neutrophil count and fibrinogen.

Differentiation between infectious and noninfectious disease and rapid initiation of accurate treatment are essential in managing diseases in the neonatal and young foal. Identification of useful inflammatory markers for these purposes is, therefore, of great importance. The aim of this study was to compare the responses of the acute phase protein serum amyloid A (SAA) with the responses of fibrinogen and total leucocyte and neutrophil counts in infectious diseases encountered in the young foal, and to assess whether SAA measurements give additional information useful in the management of these diseases. In a prospective study, foals (n = 25) showing clinical signs indicative of infectious disease were blood sampled on admission and then daily or every second day during hospitalisation. The main presenting signs were neonatal weakness (n = 9), pneumonia (n = 6) and diarrhoea (n = 10). SAA and fibrinogen concentrations on admission were higher in foals with bacterial infections (n = 8) than in foals with nonbacterial or uncertain diagnoses (n = 17). On admission, weak foals with negative blood cultures (n = 3) had normal SAA and fibrinogen concentrations and varying total leucocyte and neutrophil counts. Foals with positive blood cultures (n = 2) had markedly increased SAA, decreased or increased fibrinogen concentration and leuco- and neutropenia. Those with ambiguous blood cultures (n = 3) had moderate to markedly increased SAA concentrations and normal fibrinogen concentration, leucocyte and neutrophil counts on admission. All foals with negative or ambiguous blood cultures recovered and had normal or decreasing SAA concentration on discharge. Both foals with a positive blood culture were subjected to euthanasia. One foal born with equine herpesvirus-1 infection had moderately increased SAA and normal fibrinogen concentration and leuco- and neutropenia. Foals with Rhodococcus equi pneumonia had increased concentrations of all parameters on admission. On discharge, recovered foals had normal SAA concentrations, whereas fibrinogen and total white blood cell count and neutrophil counts were still increased. There were no consistent inflammatory changes in the parameters measured in diarrhoeic foals and there was no statistical difference between rotavirus-positive (n = 4) and -negative (n = 6) foals in this respect. The results of this investigation suggest that SAA might be an aid in the differential diagnostic procedure of neonatally weak foals and in foals with diarrhoea as the main presenting clinical sign and that SAA measurements could add information in the monitoring of treatment in Rhodococcus equi pneumonia by responding more rapidly than the markers used to date.

Gas exchange during intense exercise in Standardbreds with earlier Rhodococcus equi pneumonia.

It is not known if pulmonary function and gas exchange during exercise are altered after pyogranulomatous pneumonia caused by Rhodococcus equi infection in the foal. The aim was to evaluate whether pulmonary gas exchange during high intensity exercise was altered in mature Standardbreds with a history of R. equi pneumonia as foals. In 7 foals, R. equi pneumonia was confirmed and treated. At age 3 years, when these horses were subjected to professional training, an inclined treadmill exercise test including 4 speeds was performed. Samples were collected when a steady state in VO2 was obtained. Red cell volume, heart rate, respiratory rate, and systemic and pulmonary mean arterial pressures were measured and cardiac output calculated. Oxygen and carbon dioxide tensions in arterial and mixed venous blood were analysed. The alveolar ventilation and the alveolar-arterial oxygen tension difference were determined. Pulmonary gas exchange was assessed and the ventilation-perfusion distribution, VA/Q, was estimated by the multiple inert gas elimination technique. Ventilation-perfusion mismatch and shunt were determined and diffusion limitation calculated. The gas exchange in Standardbred trotters previously infected with R. equi and successfully treated was not compromised during intense treadmill exercise compared with reference values for healthy, fit Standardbreds. We conclude that adult Standardbreds trotters with diagnosed R. equi pneumonia as foals, can achieve an adequate gas exchange at a workload close to VO2peak.

Neutrophil functions and serum IgG in growing foals.

The aim of this study was to investigate the phagocytic and killing capacities as well as expression of CD18 of neutrophils obtained from healthy foals from birth to age 8 months. Blood was taken from 6 Standardbred foals at 7 time-points between ages 2-56 days and thereafter once a month. For comparison, cells from 16 mature horses were evaluated. Neutrophil phagocytosis of yeast cells was assessed by flow cytometry after opsonisation with mature pooled serum, autologous serum or anti-yeast IgG. The killing capacity of the neutrophils, as indicated by the oxidative burst, was monitored by chemiluminescence. Serum IgG concentration was measured by radial immunodiffusion. In addition to clinical examination, the amount of serum amyloid A and the total leucocyte count were used as markers for infection. The phagocytic ability was impaired until age 3 weeks, when autologous serum was used as opsonin. Killing capacity was also low initially but, from 3 months onwards, chemiluminescence values were equal to or higher than in mature horses. Serum IgG decreased from 10 g/l at 2 days to 5 g/l at 2 months and then increased gradually to 10 g/l at the end of the study. These findings may in part explain the increased susceptibility to bacterial infections in young horses.

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.

Flow-cytometric studies of the phagocytic capacities of equine neutrophils.

Methodological aspects of flow-cytometric evaluation of the phagocytic properties of equine neutrophils were elucidated. The kinetics of attachment and ingestion were studied, and the phagocytic process was more rapidly completed when serum-opsonized yeast cells were used than with use of IgG-opsonized yeast cells. Trypan blue was successfully used to quench fluorescence of non-ingested yeast cells. There were only minor differences in the kinetics of phagocytosis between quenched and unquenched samples, indicating that attachment is rapidly followed by ingestion. Trypan blue quenching caused loss of cells with light scattering properties of granulocytes, although this did not affect the determined frequencies of truly phagocytic neutrophils. Aggregation of yeast cells proved to be a disturbance but not an obstacle to the determination of frequencies of actively phagocytic cells. Flow cytometry is well suited for studies of phagocytosis of yeast cells by equine neutrophils, and the trypan blue quenching provides a means of eliminating false-positive events due to aggregation of yeast cells. The main advantage of the flow-cytometric method is the possibility of rapid processing of a large number of samples, making the method useful for studies of herds.