Immunofluorescent evaluation of the lower respiratory tract of healthy horses and of horses with chronic bronchiolitis

Pulmonary parenchymal tissues from 6 healthy horses and from 9 horses with chronic bronchiolitis were evaluated by use of an indirect immunofluorescent technique. In horses of both groups, the diffuse interstitial immunofluorescence was most intense for immunoglobulin (Ig)G, moderate for IgA, and minimal for IgM. Intensity of fluorescence was increased markedly in diseased lungs exposed to anti-IgA and anti-IgG. Around small bronchi and large bronchioles of healthy horses, IgA-containing cells generally were more numerous than were IgG-Fc fragment (Fc)-containing cells; in small bronchioles, however, IgA-containing cells and IgG (Fc)-containing cells were identified with approximately equal frequency. Immunoglobulin-containing cell types usually were located in the lamina propria and submucosa of airways, with occasional Ig-containing cells seen interepithelially and intraluminally; IgG (T)- and IgM-containing plasma cells were observed infrequently in the same regions. The number of Ig-containing cells became fewer as the diameter of the airway decreased; therefore, at the level of the terminal bronchiole, Ig-containing cells only were observed occasionally. This pattern of Ig-containing cell distribution was similar in horses with chronic bronchiolitis; however, horses with chronic bronchiolitis had more IgA-containing cells forming perivascular and peribronchiolar cuffs than did healthy horses. The number of IgG (Fc)-containing lymphocytes and non-Ig-stainable lymphocytes also were higher in horses with chronic bronchiolitis than in healthy horses, but the number did not approach the magnitude of that observed for IgA-containing cells. One horse had deposits of complement C3 and IgG (Fc) within alveolar septa.     

Concentrations of elastinolytic metalloproteinases in respiratory tract secretions of healthy horses and horses with chronic obstructive pulmonary disease

OBJECTIVES: To determine whether samples of tracheal epithelial lining fluid (TELF) obtained from horses have elastinolytic activity characteristic of metalloproteinases, to compare elastinolytic activity in TELF obtained from healthy horses and horses with chronic obstructive pulmonary disease (COPD), and to determine whether chemically modified tetracycline-3 (CMT-3) inhibits elastinolytic activity in TELF ANIMALS: 10 horses with COPD and 10 healthy control horses. PROCEDURE: Zymography and fluorometry were used to measure elastinolytic activity, and EDTA was used to inhibit elastinolytic activity and verify that the activity was attributable to metalloproteinases. Possible inhibition of elastinolytic activity with CMT-3 was studied in vitro. RESULTS: Elastinolytic activity was found in TELF obtained from all horses, and this activity was significantly higher in TELF obtained from horses with COPD than in TELF obtained from healthy horses. For all samples, EDTA and CMT-3 inhibited elastinolytic activity. CONCLUSIONS AND CLINICAL RELEVANCE: Elastinolytic activity is detectable in TELF obtained from horses and seems to be attributable to metalloproteinases. Elastinolytic activity in TELF is significantly inhibited by CMT-3. Elastinolytic activity in TELF can be detected by means of zymography or fluorometry. Increased elastinolytic activity may reflect destruction of pulmonary tissue in horses with COPD. Chemically modified tetracyclines such as CMT-3 may provide an additional treatment possibility for horses with COPD.     

Isolation of Streptococcus pneumoniae from the respiratory tract of horses

Streptococcus pneumoniae was isolated from nasopharyngeal swabs and tracheal washings taken from Thoroughbred horses in training at three of four separate stables that were sampled during investigations into respiratory disease. The growth of Strep pneumoniae in culture was enhanced by an environment enriched with carbon dioxide. In one stable, five of 15 horses that were sampled repeatedly were found to carry the organism for at least four months. There was an apparent association between lower respiratory tract inflammatory disease and heavy growths (10(6) to 10(8) colony forming units/ml) predominantly of Strep pneumoniae or of that organism together with large numbers of Strep zooepidemicus obtained from tracheal washings. Twelve strains of Strep pneumoniae isolated from three stables were all of capsule Type 3. Only one strain, which was of capsule Type 9, was isolated from nose and throat swabs taken from 32 staff working in one of the stables and suggested an absence of cross infection between horses and their handlers in this instance.     

Bacterial infection of the lower respiratory tract in 34 horses

OBJECTIVE: To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. PROCEDURE: Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. RESULTS: Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. CONCLUSION: There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were euthanased than from those that survived. There was an association between failure of horses to recover from pleuropneumonia and delay in diagnosis and initiation of treatment.     

Virologico-serologic studies in horses with respiratory tract diseases]

Of 1081 acute and chronically respiratory diseased as well as clinically normal horses 824 sera and 257 paired serum samples collected 1986 and 1987 were tested for antibodies against several different respiratory viruses such as influenza virus A/equi 1 and 2 (Influenza 1 a. 2), equine herpesvirus type 1/4 (EHV 1/4), mammalian reovirus type 1-3 (Reovirus 1-3), equine rhinovirus type 1 (ERV 1), equine adenovirus type 1 (EAdV 1), and equine arteritis virus (EAV). The investigations resulted in an antibody prevalence of 57.2% (Influenza 1), 59.5% (Influenza 2), 81.5% (EHV 1/4), 50.3% (Reovirus 1), 43.0% (Reovirus 2), 75.9% (Reovirus 3), 97.6% (EAdV 1), 82.5% (ERV 1) and 8.7% (EAV). With exception of EAV and EAdV 1 the ratios usually were higher in diseased animals than in clinically normal horses. Antibodies to EAV and EAdV 1 were present in all groups to almost the same amount. Of 257 horses with acute respiratory illness 3 showed a significant rise of the antibody titer against Influenza 1, 30 against Influenza 2, 54 against EHV 1/4, 1 against Reovirus 1 and 3, respectively, 11 against EAdV 1 and 26 against ERV 1.     

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. β-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 Enterobacteriaceaé 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 β-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.     

Mononuclear phagocytes of transport-stressed horses with viral respiratory tract infection

Twelve horses comprised 3 treatment groups; all horses in 2 of the groups had recently been transported and had clinical and laboratory evidence of respiratory tract infection, with equine influenza type 2 virus being the principal pathogen. Mononuclear phagocytes and other leukocytes from blood, lung, and peritoneal cavity were studied in phagocytosis and erythrocyte-antibody (EA) rosette assays. Total numbers of pulmonary alveolar macrophages were increased over control values in bronchoalveolar lavage (BAL) fluid of group 3 horses after recovery from influenza (P less than 0.02), whereas the increase in neutrophils in the fluid of those horses approached significance. Lymphocytes in BAL fluid of group 3 horses (after recovery from influenza) were in larger proportion than those in fluid of group 1 horses during acute influenza (P less than 0.05). Pulmonary alveolar macrophages of group 1 horses formed a lower percentage of EA rosettes than did those of controls (P less than 0.01) or group 3 horses (P less than 0.02). The differential counts of peritoneal macrophages and neutrophils in horses of groups 1 and 3 were virtually identical at the first collection, but differed from controls at the second collection 4 weeks later; peritoneal macrophages were reduced (P less than 0.01), whereas peritoneal neutrophils were increased (P less than 0.01). Peritoneal macrophages and peritoneal neutrophils of horses with acute influenza were phagocytic in larger proportion than were those in controls at both collection times (P less than 0.01 and P less than 0.01 for peritoneal macrophages, and P less than 0.01 and P less than 0.05 for peritoneal neutrophils, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of bacitracin on microbial functions in the gastrointestinal tract of horses

This study investigated the influence of zinc bacitracin on the intestinal flora of horses. The functionally active intestinal flora was examined in 6 horses during treatment with zinc bacitracin. Utilising gas chromatography, spectrophotometry, gel electrophoresis and paper chromatography, samples were analysed on biochemical markers reflecting the action of parts of the intestinal flora. The following 5 flora-related functions were studied in faecal samples and intestinal samples from different sections of the hindgut: conversion of cholesterol to coprostanol and of bilirubin to urobilinogens, degradation of mucin and of beta-aspartylglycine and inactivation of tryptic activity. Conversion to coprostanol, conversion to urobilinogens and degradation of mucin were affected by treatment of zinc bacitracin and conversion to coprostanol was most sensitive. All functions were normalised in a short time, in contrast to man and rats. Differences in environmental exposures are probably the reason for a more rapid normalisation of the intestinal flora functions in horses.     

Isolation of mycoplasmas from the respiratory tract of horses in Australia.

Mycoplasmas were isolated from two of 43 nasal swabs taken from live horses, and from one of 28 tracheal swabs taken from slaughtered horses. The slaughtered horse that yielded mycoplasmas had no gross pathological changes in the respiratory tract, but the nasal isolations were made from horses with rhinitis. The three mycoplasmas could be distinguished by cultural characteristics, and probably they represent three different species.     

Cytology of the lower respiratory tract in horses: An updated review

The diagnosis of respiratory diseases in horses can be difficult due to the nonspecific nature of the clinical signs, which overlap between the different diseases. Cytological examination of the respiratory secretions is a helpful diagnostic tool that, when combined with clinical features and other diagnostic techniques, has the potential to improve diagnostic accuracy significantly. This review article should help readers to become familiar with the cytology of the normal respiratory tract and with the cytopathological abnormalities associated with common respiratory disorders in horses.     

Morphine administration in horses anaesthetized for upper respiratory tract surgery

OBJECTIVE: To determine the effect of morphine administration on commonly monitored cardio-respiratory variables and recovery quality in horses undergoing anaesthesia and surgery. STUDY DESIGN: Prospective, randomized clinical study. ANIMALS: Thirty-eight thoroughbred horses, 32 geldings and six mares, 3-13 years old, weighing 411-600 kg. MATERIALS AND METHODS: A standard anaesthetic technique was used. Twenty minutes after induction of anaesthesia horses received 0.1 mg kg(-1) (0.1 m) or 0.2 mg kg(-1) (0.2 m) morphine by intravenous injection. A control group did not receive morphine. Heart rate, respiratory rate (fr), mean arterial pressure (MAP) and blood gases were measured before morphine administration and every 10 minutes thereafter. Horses were positioned for 35 minutes in right lateral recumbency for tension palatoplasty by cautery and were then moved into dorsal recumbency for additional intraluminal surgery comprising one or more of aryepiglottic fold resection, sub-epiglottal mucosal resection, ventriculectomy and cordectomy. A subjective recovery score from 0 (worst) to 5 (best) was assigned by a single observer who was unaware of treatment group. Two-way repeated measures anova, one-way anova, Kruskal-Wallis test, Mann-Whitney test, Pearson and Spearman correlation coefficients, and chi-squared tests were used to analyse the data where appropriate. RESULTS: Arterial partial pressure of oxygen (PaO(2)) decreased significantly over time and was significantly lower in horses that received morphine. One horse in the control group and two horses in each of the morphine groups had a PaO(2) <13 kPa. No other significant cardiopulmonary effects were detected. Recovery scores [median (range)] were higher in morphine recipients: 4 (2-5) in 0.1 m, 4 (3-5) in 0.2 m compared with 3 (2-4) in the control group. CONCLUSIONS AND CLINICAL RELEVANCE: The lower PaO(2) in morphine recipients did not appear to be of clinical significance in healthy horses because the number of horses with a low PaO(2) was similar between groups. The quality of recovery was significantly better in morphine recipients. These results indicate that morphine may be considered for use in clinical cases although further work is required to assess the analgesic properties of the drug in this species.     

Influence of systemic fluoroquinolone administration on the presence of Pasteurella multocida in the upper respiratory tract of clinically healthy calves

The influence of enrofloxacin administration (5 mg/kg) for five consecutive days on the occurrence of Pasteurella multocida in the upper respiratory tract of two healthy calves was monitored over a 10-day period. From nasal swabs of two additional healthy control calves, which received a placebo saline administration, P. multocida was isolated throughout the study period. In the enrofloxacin treated calves, P. multocida was not demonstrated in the nasopharynx from 48 h after the first injection until two days after the last administration, when P. multocida reappeared and proved to be clonal in nature to the original isolates. During the experiment, no change in minimal inhibitory concentration for enrofloxacin of the P. multocida isolates was detected (MIC ≤ 0.015 μg/mL). Enrofloxacin concentrations were determined in the plasma by a high-performance liquid chromatography method with fluorescence detection. The PK/PD indices AUC/MIC and C_max/MIC ratio were calculated and found to be 1157.7 and 129.8, respectively. Remarkably, the respiratory pathogen Arcanobacterium pyogenes became the predominant recovered organism in the nasopharynx of one animal following enrofloxacin therapy throughout the remaining of the experiment.     

Plasma concentrations of endothelin-like immunoreactivity in healthy horses and horses with naturally acquired gastrointestinal tract disorders

OBJECTIVE: To compare plasma endothelin (ET)- like immunoreactivity between healthy horses and those with naturally acquired gastrointestinal tract disorders. ANIMALS: 29 healthy horses and 142 horses with gastrointestinal tract disorders. PROCEDURE: Blood samples were collected from healthy horses and from horses with gastrointestinal tract disorders prior to treatment. Magnitude and duration of abnormal clinical signs were recorded, and clinical variables were assessed via thorough physical examinations. Plasma concentrations of ET-like immunoreactivity were measured by use of a radioimmunoassay for human endothelin-1, and CBC and plasma biochemical analyses were performed. RESULTS: Plasma ET-like immunoreactivity concentration was significantly increased in horses with gastrointestinal tract disorders, compared with healthy horses. Median plasma concentration of ET-like immunoreactivity was 1.80 pg/ml (range, 1.09 to 3.2 pg/ml) in healthy horses. Plasma ET-like immunoreactivity was greatest in horses with strangulating large-intestinal obstruction (median, 10.02 pg/ml; range, 3.8 to 22.62 pg/ml), peritonitis (9.19 pg/ml; 789 to 25.83 pg/ml), and enterocolitis (8.89 pg/mI; 6.30 to 18.36 pg/ml). Concentration of ET-like immunoreactivity was significantly associated with survival, PCV, and duration of signs of pain. However, correlations for associations with PCV and duration of pain were low. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with gastrointestinal tract disorders have increased plasma concentrations of ET-like immunoreactivity, compared with healthy horses. The greatest values were detected in horses with large-intestinal strangulating obstructions, peritonitis, and enterocolitis. This suggests a potential involvement of ET in the pathogenesis of certain gastrointestinal tract disorders in horses.     

Influence of clenbuterol on mucociliary transport in healthy horses and horses with chronic obstructive pulmonary disease

The mucociliary tracheal transport rate and clearance index were measured in 16 healthy horses and 16 horses suffering from chronic obstructive pulmonary disease. A marker, Indian ink in syrup, was placed in the mucus layer of the interior wall of the trachea, just in front of the carina, and its displacement was measured after 30 and 60 minutes. The same procedure was repeated 30 minutes after the intravenous administration of 0.8 microgram clenbuterol/kg bodyweight. In the diseased horses the mucociliary transport rate was significantly lower than in the healthy horses. After clenbuterol administration there was a significant increase in the transport rate and a similar increase in the clearance index. Sixty and 90 minutes after clenbuterol there was a significant increase of mucus secretion in both the healthy and diseased horses. Measurements in five healthy horses and five diseased horses on two different days gave similar results, showing that the technique provided reproducible data.     

The effect of respiratory activity on the QRS complex in the ECG in clinically healthy horses and horses with chronic lung diseases

Interrelations between respiration and changes in amplitudes of the QRS-complex of the ECG of horses were investigated. The value of the R-vector and both its components Rx and Ry decrease during inspiration and increase in the expiration phase. It means, that the R-waves in leads from cranial to caudal (y, II, aVF) and from right to left (I) become smaller with inspiration and larger with expiration. The difference of the amplitudes may be important, especially in horses with respiratory problems, so that it has to be noted when evaluating electrocardiograms. For that purpose it is not necessary to draw respiratory curves. It is proposed to evaluate the heart-cycle with the largest R-wave (expiration) only when there are differences in the R-amplitudes. Pulmonary air content is discussed as a possible cause.     

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

SUMMARY: 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 10⁹ 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.