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Biomedical subjects

S L Raidal

Publications and source records attributed to S L Raidal.

At least 19 recordsLinked to original sources

Detection of respiratory herpesviruses in foals and adult horses determined by nested multiplex PCR.

A nested multiplex PCR was developed as a rapid (<12h), sensitive test for the simultaneous identification of equine herpesviruses (EHV1, EHV4, EHV2 and EHV5) in clinical samples from horses. Peripheral blood and nasal swab (NS) samples from 205 weanling Thoroughbred foals on 6 different studs over 3 consecutive seasons and from 92 adult horses without clinical signs of respiratory disease were examined using direct multiplex PCR of clinical samples (direct PCR) and conventional cell culture with differentiation of EHV in cell cultures by multiplex PCR. Multiplex PCR proved a sensitive and specific technique for the detection of EHV in cell culture and clinical samples. The technique described appeared equally sensitive as one using a single set of primers for individual EHV but reduced labour and reagent costs. Cell cultures showing cytopathic effect (CPE) were always positive for EHV on PCR. EHV were also detected by multiplex PCR in 11 samples which failed to show CPE. By a combination of multiplex PCR and cell culture or direct multiplex PCR, the presence of up to three EHV in the same sample was detected. Overall, EHV5 was detected by direct multiplex PCR of peripheral blood mononuclear cells (PBMC) and/or NS samples from 78% of foals and 47% of adult horses. Repeated sampling or cell culture in combination with multiplex PCR and with the incorporation of IL-2 in culture medium increased the sensitivity for detection of EHV in PBMC and demonstrated that EHV5 DNA could be identified in PBMC from 89% of foals and 100% of adult horses. EHV2 was identified from approximately 30% of foals, but was more frequently identified in samples from 17 foals with mild respiratory disease and was isolated infrequently from adult horses. EHV1 and EHV4 were identified uncommonly in any population in the current study.

Animals↗

Inoculation of mares and very young foals with EHV-1 glycoproteins D and B reduces virus shedding following respiratory challenge with EHV-1.

We have previously demonstrated that intramuscular inoculation of EHV-1 glycoprotein D (gD) and glycoprotein B (gB) produced by a recombinant baculovirus and formulated with the adjuvant Iscomatrix elicited virus-neutralizing antibody and gD- and gB-specific ELISA antibody in adult horses. In this study, 14 mares and their very young foals were inoculated with a combination of baculovirus-expressed EHV-1 gD and EHV-1 gB (EHV-1 gDBr) and challenged with a respiratory strain of EHV-1. Following experimental challenge, inoculated mares and foals shed virus in nasal secretions on significantly fewer occasions compared to uninoculated mares and foals. Uninoculated foals born from inoculated mares were no more protected against experimental challenge than uninoculated foals born from uninoculated mares. The results suggest that it is indeed possible to induce partial protection in very young foals through vaccination, and while the inoculation did not prevent infection, it did reduce the frequency of viral shedding with the potential to thereby reduce the risk and prevalence of infection in a herd situation.

Animals↗

Pharmacokinetics of potassium bromide in adult horses.

OBJECTIVE: To determine the pharmacokinetics of potassium bromide (KBr) in horses after single and multiple oral doses. ANIMALS: Twelve adult Standardbred and Thoroughbred mares. PROCEDURE: Horses were randomly assigned to two treatment groups. Group 1 horses were given a single oral dose of 120 mg/kg potassium bromide. Part 2 of the study evaluated a loading dose of 120 mg/kg KBr daily by stomach tube for 5 days, followed by 40 mg/kg daily in feed for 7 days. Serum concentrations of KBr were measured to construct concentration versus time curves and to calculate pharmacokinetic parameters. Treated horses were monitored twice daily by clinical examination. Serum concentrations of sodium, potassium and chloride ions and partial pressures of venous blood gases were determined. RESULTS: Maximum mean serum concentration following a single dose of KBr (120 mg/kg) was 423 +/- 22 microg/mL and the mean elimination half-life was 75 +/- 14 h. Repeated administration of a loading dose of KBr (120 mg/kg once daily for 5 d) gave a maximum serum concentration 1639 +/- 156 microg/mL. The administration of lower, maintenance doses (40 mg/kg once daily) was associated with decreased serum bromide concentrations, which plateaued at approximately 1000 microg/mL. Administration of KBr was associated with significant but transient changes in serum potassium and sodium concentrations, and possible changes in base excess and plasma bicarbonate concentrations. High serum concentrations of bromide were associated with an apparent increase in serum chloride concentrations, when measured on an ion specific electrode. CONCLUSIONS: and clinical relevance Loading doses of 120 mg/kg daily over 5 d and maintenance doses of approximately 90 mg/kg of KBr administered once daily resulted in serum bromide concentrations consistent with therapeutic efficacy for the management of seizures in other species. The clinical efficacy of this agent as an anticonvulsant medication and/or calmative in horses warrants further investigation.

Administration, Oral↗

A comparison of foal and adult horse neutrophil function using flow cytometric techniques.

Flow cytometric assays were used to compare phagocytic and oxidative burst activity of neutrophils from healthy foals less than 7 days of age with the activity of cells from healthy adult horses. The phagocytosis of Staphylococcus aureus by foal neutrophils was less than that observed for adult neutrophils when autologous serum was used as the source of opsonins in the assay. The use of adult serum did not significantly improve the ability of foal neutrophils to attach bacteria. The oxidative burst activity of foal neutrophils was equivalent to that of adult cells. However, when serum or plasma was incorporated into the oxidative burst assay, foal neutrophils demonstrated greatly reduced autofluorescence and a suppressed response to phorbol myristate acetate (PMA), relative to that demonstrated by adult cells. These results suggest that peripheral blood neutrophils from foals have a reduced ability to phagocytose bacteria relative to that exhibited by adult horse neutrophils and that the oxidative burst activity of foal neutrophils is down-regulated in response to an unidentified serum factor(s). Such changes may contribute to the increased susceptibility of foals to septic disease.

Age Factors↗

Effects of training on resting peripheral blood and BAL-derived leucocyte function in horses.

In this study, the effects of prolonged, high intensity training on aspects of peripheral blood and bronchoalveolar lavage (BAL)-derived leucocyte function were evaluated in 8 horses. All horses undertook a 7 week endurance training programme, followed by 5 weeks of high intensity training (HIT). Thereafter, horses were divided into control (C) and overtraining (OT) groups. The frequency and intensity of training were increased more substantially for horses in the OT group. Training was terminated in week 32 when horses in the OT group demonstrated a significant performance reduction. Peripheral blood and BAL samples were collected from 4 horses in C and OT groups in training weeks 7, 11, 14, 18, 22, 28 and 32. Flow cytometric techniques were used to assess phagocytosis by peripheral blood neutrophils and pulmonary alveolar macrophages (PAM), and oxidative burst activity of neutrophils, PAM, peripheral blood and BAL-derived lymphocytes. Peripheral blood neutrophil phagocytosis (internalisation) increased during the initial HIT period and decreased from week 16 when the training workload was increased for both groups. The oxidative burst activity of peripheral blood neutrophils and lymphocytes similarly increased and then decreased in response to training. The oxidative burst activity of PAM was reduced towards the end of the overtraining phase of the programme. Pulmonary alveolar macrophage phagocytosis and oxidative burst activity of BAL-derived lymphocytes demonstrated no change throughout the course of the study. There was no difference in results obtained from C or OT group horses, suggesting that protracted HIT, rather than overtraining, was associated with impaired cell function. The detrimental effects observed in peripheral blood neitrophil and PAM function may indicate impaired nonspecific immunity which may adversely affect the health and performance of horses undergoing protracted periods of intense training.

Animals↗

Effect of single bouts of moderate and high intensity exercise and training on equine peripheral blood neutrophil function.

The effects of single bouts of moderate (30 to 40 per cent VO(2)max) and high (115 per cent VO(2)max) intensity exercise on equine peripheral blood leucocyte function were evaluated by determining neutrophil phagocytosis and oxidative burst activity before and after treadmill exercise and training. Prior to all exercise tests, the possible effect of diurnal variation was evaluated in samples obtained from four resting horses. Subsequently eight horses underwent moderate and high intensity exercise protocols and then commenced a 17-week training period. High intensity exercise tests were repeated in week 10, after 7 weeks of endurance training, and in week 17, after a further 6 weeks of high intensity training. Time of sampling had a significant effect on neutrophil function for resting, untrained horses. Prior to training, moderate intensity exercise was associated with improved neutrophil phagocytosis and oxidative burst activity. High intensity exercise was associated with transient impairment of these responses. A similar reduction was not demonstrable following high intensity exercise in weeks 10 or 17 of training. Neutrophil function in week 17 was suppressed at all sampling times relative to results obtained in week 10, suggesting that high intensity training may have been associated with a general reduction in neutrophil function.

Animals↗

The effect of high intensity exercise on the functional capacity of equine pulmonary alveolar macrophages and BAL-derived lymphocytes.

The effect of strenuous exercise on the functional capacity of pulmonary alveolar macrophages (PAM) and bronchoalveolar lavage-derived lymphocytes was determined in eight horses prior to and after 7 weeks of training. Strenuous exercise had no effect on the total cell count or the percentage of live cells in bronchoalveolar lavage (BAL) samples prior to or following training. However, training was associated with a significant increase in the total cell count of pre-exercise BAL samples and a significant reduction in the percentage of live cells in post-exercise samples. Strenuous exercise was associated with impaired phagocytosis by PAM after 7 weeks of training but had no effect on similar samples obtained from untrained horses. The oxidative burst activity of PAM was significantly increased following strenuous exercise for both untrained and trained horses. BAL -derived lymphocyte oxidative burst was similarly affected following training. These results suggest that strenuous exercise and training may influence pulmonary immune cell function.

Animals↗

The flow cytometric evaluation of phagocytosis by equine peripheral blood neutrophils and pulmonary alveolar macrophages.

Flow cytometry was used to assess the phagocytosis of fluorescent-labelled bacteria by equine peripheral blood neutrophils and pulmonary alveolar macrophages. Cell populations were prepared from venous blood following ammonium chloride lysis and from washed bronchoalveolar lavage derived samples. Discrete clusters of cells, corresponding to different leucocyte groups, were readily identified on the basis of differing light scattering properties and could thus be discriminated, negating the need for prior cell separation. Cells able to associate with fluorescent-labelled bacteria (by attachment to the cell membrane or by internalization within the cell) acquired increased fluorescence and were readily differentiated from cells unable to interact with bacteria. The fluorescence of bacteria attached to the cell surface was quenched by the addition of trypan blue or counterstained by the addition of ethidium bromide to the assay, thus permitting identification of cells which were able to internalize bacteria.

Animals↗

Flow cytometric determination of oxidative burst activity of equine peripheral blood and bronchoalveolar lavage-derived leucocytes.

Flow cytometric techniques were developed for the evaluation of oxidative burst activity in equine peripheral blood neutrophils and lymphocytes, as well as bronchoalveolar lavage derived pulmonary alveolar macrophages and lymphocytes. The oxidation of dichlorofluorescin was measured by the increased fluorescence of cells stimulated with phorbol myristate acetate or a variety of other stimulants. Flow cytometry was a suitable method for the evaluation of the intracellular oxidation in all cell populations evaluated. Analysis was rapid and cell separation before analysis was not required. Heterogenous cell populations with differing responsiveness to phorbol myristate acetate stimulated oxidative burst were identified in peripheral blood neutrophil and alveolar macrophage populations. The current study characterizes flow cytometric techniques for the evaluation of oxidative burst activity in equine peripheral blood and bronchoalveolar lavage-derived leucocytes.

Animals↗

Antibiotic prophylaxis of lower respiratory tract contamination in horses confined with head elevation for 24 or 48 hours.

OBJECTIVE: To evaluate the administration of procaine penicillin prior to or during confinement with head elevation as a means of reducing the associated accumulation of inflammatory lower respiratory tract secretions and increased numbers of bacteria within the lower respiratory tract of confined horses. DESIGN AND PROCEDURE: Two experiments were conducted to evaluate the efficacy of different dose rates and dosing frequencies. In experiment A a single low dose (15,000 IU/kg) of procaine penicillin was administered to four horses immediately prior to confinement with head elevation for 48 hours. The systemic leucocyte response, gross and cytologic characteristics of transtracheal aspirate and bacterial numbers in lower respiratory tract samples were compared with corresponding samples from two horses confined with heads elevated but not given penicillin. The efficacy of higher dose rates (20,000 IU/kg and 40,000 IU/kg) given before and during confinement with heads elevated for 24 hours was evaluated in experiment B. RESULTS: Treatment with procaine penicillin had no effect on the systemic leucocyte response or on the accumulation of inflammatory lower respiratory tract secretions at any of the dosing schedules evaluated. The number of bacteria isolated from trans-tracheal samples was reduced at 12 hours for treated horses in experiment A and at 24 hours for experiment B. beta-haemolytic Streptococcus spp were not isolated from treated horses in either experiment. Bacterial species isolated from treated horses were predominantly Pasteurella and/or Actinobacillus spp, however, members of the family Enterobacteriaceae and a Staphylococcus sp were isolated from treated horses. One treated horse in experiment A developed clinically apparent pulmonary disease. CONCLUSIONS: The prophylactic administration of penicillin before or during confinement did not reliably reduce bacterial numbers or prevent the accumulation of purulent lower respiratory tract secretions in horses confined with their heads elevated. Numbers of beta-haemolytic Streptococcus spp were reduced following treatment, suggesting that the repeated administration of procaine penicillin may have some merit as part of a strategy to prevent transport-associated respiratory disease. However, methods directed at minimising the duration of confinement with head elevation, augmentation of the clearance of accumulated secretions and prompt identification of animals in which airway inflammation has extended to the pulmonary parenchyma remain the best ways of minimising transport-associated respiratory disease.

Actinobacillus↗

Effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function.

OBJECTIVE: To evaluate the effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function in horses and to compare results from transported horses with those obtained in earlier experiments from horses confined with heads elevated. DESIGN: A prospective study. PROCEDURE: Six horses were transported by road for 12 h. Clinical and haematological examination, transtracheal aspiration and cell function studies were conducted before and after transportation. Results obtained after transportation were compared to pre-transportation values. RESULTS: After transportation, peripheral blood leucocyte and neutrophil numbers were increased and rectal temperatures were evaluated. Transtracheal aspirates showed an accumulation of purulent respiratory tract secretions with increased numbers of bacteria, particularly beta-haemolytic Streptococcus spp and members of the Pasteurellaceae family. Three horses also had increased numbers of bacteria from the Enterobacteriaceae family relative to corresponding samples from earlier studies. Phagocytosis by peripheral blood neutrophils was significantly reduced, while the oxidative burst activity of peripheral blood leucocytes was either unchanged or enhanced. CLINICAL IMPLICATIONS: Bacterial contamination of the lower respiratory tract occurs as a routine consequence of transportation of horses and is likely to be an important determinant in the development of transport-associated respiratory disease. Inflammatory airway secretions and increased numbers of bacteria were rapidly cleared, without clinical evidence of significant pulmonary disease and without additional treatment, in normal horses that were allowed to lower their heads after transportation. Peripheral blood neutrophilia and a reduction in neutrophil phagocytic function were evident for at least 36 h after transportation, suggesting that horses may require a number of days to recover from the stress of transportation. As the potential role of bacteria from the Enterobacteriaceae family in the development of transport-associated respiratory disease has not been elucidated, horses which develop clinical disease following transportation should undergo thorough bacteriological investigation to ensure appropriate treatment.

Animals↗

Effects of posture and accumulated airway secretions on tracheal mucociliary transport in the horse.

Tracheal mucociliary clearance was determined in horses by measuring the rostrad transport of the radiopharmaceutical 99mtechnetium-sulphur colloid following deposition on the tracheal epithelium by intratracheal injection. The effects of head position (head elevated to normal standing position vs head lowered) and of accumulated purulent secretions on tracheal mucociliary clearance were evaluated for the first time in the horse. In normal horses tracheal mucociliary clearance was greatly accelerated by lowering the head so that the cranial trachea was lower than the caudal trachea. Horses confined with their heads elevated for 24 hours developed an accumulation of purulent airway secretions (and associated increased numbers of bacteria) in the lower respiratory tract and showed a decrease in tracheal mucociliary clearance when compared with their previously measured rate when the lower airway contained only normal secretions. These findings have implications for management practices where horses are prevented from lowering their heads, such as transportation and cross-tying, which may therefore contribute to lower respiratory tract disease in horses.

Animals↗

The incidence and consequences of failure of passive transfer of immunity on a thoroughbred breeding farm.

Circulating IgG concentration was determined between 12 and 24 hours after birth for 323 foals born on a Thoroughbred breeding farm over 3 consecutive years. The incidence of failure of passive transfer (FPT) of maternal immunoglobulins (foal circulating IgG concentration < 8 g/L) was found to be 9.6%. Foals born late in the season (October to December) were found to be at increased risk for the development of FPT. The degree of assistance required at parturition and the presence of a periparturient problem in the mare or foal also significantly influenced the subsequent incidence of FPT. Passive immune status significantly influenced the likelihood of foals developing septic illness (joint ill, septicaemia, pneumonia) in the first month of life, but had no significant effect on the development of diarrhoea or Rhodococcus equi pneumonia. The results of the current study support the value of routine monitoring of passive immune status and the early speculative treatment of foals considered to be at risk for the development of FPT.

Animals↗

A retrospective evaluation of the surgical management of equine carpal injury.

Records of 220 thoroughbred horses presented to the Randwick Equine Centre or the University of Queensland Veterinary Teaching Hospital for surgical management of carpal injury were reviewed. Details of racing performance were obtained, enabling evaluation of racing success following surgery. Age and sex matched control horses not known to have suffered carpal injury were selected from the Australian Stud Book and the Australian Racehorse Register. Control and treated populations were compared in terms of overall career racing success. Radiographs or xeroradiographs from 198 horses were available for evaluation. The dorsomedial aspects of the distal articular surface of the radial carpal bone and the proximal articular surface of the third carpal bone were the most commonly observed locations of osteochondral fracture. Bilateral carpal injury was found to be sufficiently common to warrant routine radiographic examination of both limbs. The dorsolareral-palmaromedial oblique, flexed lateral and dorsoproximal-dorsodistal oblique (skyline) projections were the most useful views for evaluation of carpal degenerative joint disease. Surgical removal of osteochondral fractures by arthrotomy or arthroscopy was found to be a suitable method of treatment. Arthroscopy was associated with a significantly shorter convalescent period than arthrotomy. Seventy-six percent of the treated population returned to racing following surgery. Average earnings following surgery were $20,000. However, the median value was only $1400 and 48% of the treated population failed to earn more than $1000. Male horses in both the treated and control groups performed significantly better than females. Treated horses performed significantly better than the randomly selected control population.

Animals↗

Inflammation and increased numbers of bacteria in the lower respiratory tract of horses within 6 to 12 hours of confinement with the head elevated.

Confinement of horses with their heads elevated for periods up to 24 hours was used to evaluate the extent and the effects of bacterial contamination of the equine lower respiratory tract. Significant (P < 0.05) increases in bacterial numbers (up to 10(9) colony forming units/mL in transtracheal aspirate derived samples) occurred within 6 or 12 hours in most horses. Pasteurella/Actinobacillus spp and Streptococcus spp were most commonly isolated. Lowering of the head for 30 minutes every 6 hours to facilitate postural drainage did not prevent multiplication of organisms to levels equivalent to those achieved by horses where the head was elevated for 24 hours. When horses were released from confinement and heads were no longer maintained in an elevated position, clearance of accumulated secretions and bacteria occurred within 8 to 12 hours. Thus, confinement with the head elevated resulted in significant bacterial contamination and multiplication within the lower respiratory tract during a period often encountered in routine management procedures, such as transportation. The clearance of accumulated secretions occurred over a prolonged period after release from such confinement.

Animals↗