Respiratory-induced variability of pulmonary arterial pressure measurements in cattle

The purpose of this study was to investigate the variability of the peak systolic (PAPs) and the end diastolic (PAPd) pulmonary arterial pressures induced by intrapleural pressure changes in cattle.The pleural pressure (Ppl), the electrocardiogram and the pulmonary arterial pressure (PAP) were simultaneously recorded in five healthy calves under three different conditions, i.e. normoxia (N), normoxia with an added airflow resistance (R) and hypoxia (H). PAPs, PAPd and their corresponding transmural pressures were measured and averaged over 10 successive regular cardiac cycles. The maximum Ppl changes (maxPpl) were measured on the same tracings. The variance and coefficients of variation were calculated for each set of vascular measurements.MaxPpl was significantly increased with regard to N values during R and H conditions. This increase in maxPpl induced a simultaneous rise in the variability of PAP measurements, while in each condition, this variability was greatly lowered by use of the corresponding transmural pressure.It was concluded that, in calves with high maxPpl, the influence of respiration on PAP becomes considerable. In such cases, the use of transmural pressures rather than luminal pressures can greatly reduce the variability of these pulmonary pressure measurements and therefore increase their sensitivity.     

Synergistic action of E. coli endotoxin and Pasteurella multocida type A for the induction of bronchopneumonia in pigs

This study aimed to investigate whether Escherichia coli endotoxin (LPS) may predispose the lung to an infection with Pasteurella multocida type A (Pma) and to determine the LPS concentration needed to reproduce clinical signs of bronchopneumonia. Twenty-four hours before inoculating Pma or sterile growth medium, piglets were tracheally instilled with 10, 100 or 400 microg/kg LPS. Cough, body temperature, daily weight gain (DWG) bronchoalveolar lavage fluid (BALF) cells and volume of pneumonic lung were measured. Changes in breathing pattern (Penh) were assessed by whole body barometric plethysmography. No significant changes were observed in Pma-treated or in control animals. Each LPS doses induced DWG reduction while the higher generated a severe subacute interstitial pneumonia causing hyperthermia and an increase in Penh. The combination of the lower LPS doses with Pma produced an asymptomatic bronchopneumonia leading to DWG reduction, rise in Penh and an increase in BALF macrophages and neutrophils. With 400 microg/kg LPS, Pma worsened the inflammatory process as illustrated by cough, hyperthermia, major DWG reduction and by a greater Penh response. Lung lesions consisted of severe exudative bronchopneumonia. We concluded that LPS may negatively influence growth, predispose to persisting lung inflammatory process and promote Pma infection depending on the dose previously administered.     

Pulmonary and systemic effects of inhaled endotoxin in control and heaves horses

To investigate whether inhaled endotoxin contributes to airway inflammation and dysfunction in stabled horses, control (n = 6) and asymptomatic heaves (previously termed chronic obstructive pulmonary disease)-susceptible (n = 7) horses were given inhalation challenges with 20, 200 and 2,000 microg of soluble Salmonella typhimurium Ra60 lipopolysaccharide (LPS). LPS inhalation induced a dose-dependent neutrophilic airway inflammatory response in both groups. Inhalation with 2,000 microg of LPS also induced detectable lung dysfunction in the heaves group. LPS inhalation did not alter clinical score, tracheal secretion volume or airway reactivity in either group. The no-response thresholds were lower for the heaves group (<20 microg for airway inflammation; 200 to 2,000 microg for lung dysfunction) than for the control group (20 to 200 microg for airway inflammation; >2,000 microg for lung dysfunction). To enable comparison of these threshold levels with airborne endotoxin concentrations in stables, horses also received a 5 h duration hay/straw challenge, during which the total and respirable airborne endotoxin concentrations were determined. Comparison of the effects of acute LPS inhalation and hay/straw challenges suggest that inhaled endotoxin is not the sole cause of heaves. However, it is likely that it contributes to airway inflammation, both in heaves horses in concert with other inhalants, and in normal horses when they are exposed to high levels in poor stable environments.     

Histamine inhalation challenge in normal horses and in horses with small airway disease

A histamine inhalation challenge (HIC) procedure was developed to assess hyperreactive states in horses. Following clinical evaluation, percutaneous lung biopsies were performed on nine light breed mares aged 6 to 15 years. Five horses, with normal small airways, were classified as group A and four subjects with small airway disease (SAD) lesions formed group B. Pulmonary mechanics parameters were monitored following an aerosol of 0.9% saline and every 5 min for up to 30 min after HIC with 0.5% w/v of histamine diphosphate, administered through a face mask for 2.5 min. Tidal volume (VT) and airflow (V) values were obtained with a pneumotachograph. Transpulmonary pressure (delta Ppl) was measured by the esophageal balloon catheter method. Dynamic compliance (Cdyn), total pulmonary resistance (RL), end expiratory work of breathing (EEW) and respiratory rate (f) were calculated by a pulmonary mechanics computer. Group A horses had increases in RL, and decreases in Cdyn whereas horses in group B were hyperreactive and showed greater changes in EEW, Cdyn, and delta Ppl but with a relatively lower variation of RL. One horse in clinical remission from SAD, but with a high biopsy score (group B), and one clinically normal horse belonging to group A showed marked hyperreactivity as shown by increases in EEW, maximum change in delta Ppl and RL and decreases in Cdyn. These results suggest that the HIC described can be used as a method to investigate airway hyperreactivity and SAD in horses.     

Airway responses to histamine aerosol in clinically normal foods.

OBJECTIVE: To describe the spectrum of nonspecific airway reactivity in a group of clinically normal foals. ANIMALS: 12 clinically normal mixed-breed foals, 48 to 92 days old, without history of clinical lung disease. PROCEDURE: Nonspecific airway reactivity was determined by measuring the extent of changes in dynamic compliance during nebulization of incrementally increasing concentrations of histamine aerosol. Degree of airway reactivity was expressed as the dose of histamine that evoked a decrease in dynamic compliance (Cdyn) to 65% of the after saline nebulization value (PC65Cdyn) or increase in pulmonary resistance (R(L)) to 135% of baseline (PC135R(L)). RESULTS: In all foals, it was possible to induce a decrease in Cdyn in dose-dependent manner to < or = 65% of baseline. Response of foals in terms of R(L) was more erratic, and, in 1 foal, R(L) decreased after histamine exposure. Mean+/-SD PC65Cdyn was 5.43+/-1.74 (range, 0.77 to 19.56) mg/ml, and mean PC135R(L) was 3.34+/-1.52 (range, -0.749 to 17.35) mg/ml. Body weight was not correlated to baseline Cdyn, R(L), PC65Cdyn, or PC135R(L). CONCLUSIONS AND CLINICAL RELEVANCE: Clinically normal foals had a wide range of airway reactivity, which may contribute to variation in clinical responses of foals to otherwise similar stimuli, such as infection, inflammation, and challenge exposure with environmental irritants.     

Influence of breathing pattern and lung inflation on impulse oscillometry measurements in horses

The objective of this paper was to determine if changes in ventilation patterns could influence the outcome of respiratory function measurements performed with our impulse oscillometry system (IOS) in horses. In a first study, IOS tests were performed in vitro on six isolated equine lungs. Lung inflation levels were controlled by modifying depressurisation inside an artificial thorax and different ventilation patterns were imposed. In a second in vivo study, transient variations in breathing pattern were evaluated both with the IOS and a current reference technique (CRT) in five healthy mature horses after an intravenous (i.v.) injection of lobeline hydrochloride. In both studies, respiratory rate (RR, range: 7-42 breaths/min.) and tidal volume (V(T), range: 0.4-25 L) had minor or no influence on IOS parameters. The influence of lung inflation, most marked for resistance at 5 Hz (R(5 Hz)), was limited for the considered physiological range. In vivo, statistical models indicated that maximal changes in pleural pressure (Max Delta Ppl) and peak flows were the main determinants of the variability of the resistance (R(rs)) and the reactance (X(rs)) of the respiratory system. The fourfold increase in baseline Max Delta Ppl and peak flows obtained during hyperpnoea caused a significant increase in R(rs) at 5 and 10 Hz and a decrease in X(rs) at all frequencies. We conclude that IOS parameters are not influenced by tachypnoea, but will reflect alterations in respiratory mechanics caused by hyperpnoeic breathing.     

Airway responsiveness measured by barometric plethysmography in guinea pigs

Barometric plethysmography has become an increasingly used method to indirectly measure respiratory function in unrestrained freely-moving animals. This technique has been criticized because of physiological uncertainty of its major index, the enhanced pause (Penh). Moreover, a recent study raises concerns that during histamine challenges part of the Penh response could be produced by upper airways (nasal) responses. In this study we compared airway responsiveness measured by barometric plethysmography and total lung resistance (RL) in guinea pigs, and evaluated the role of upper airways during Penh measurement. Our results showed that intravenous acetylcholine or histamine caused a dose-dependent increase of the Penh values in non-anesthetized guinea pigs, which were correlated with RL values obtained in separate groups of anesthetized animals. In anesthetized but spontaneously breathing guinea pigs intravenous acetylcholine or histamine also produced a dose-dependent increment of Penh, which was similar regardless if guinea pigs breathed through the nose or through a tracheal tube. Our results suggest that, independently of the physiological meaning of Penh, this index seems to be a useful indirect measurement for evaluating airway responsiveness to intravenous agonists in guinea pigs, and that nasal passage seems not to be involved in this measurement.     

Tracheobronchial mucus viscoelasticity during environmental challenge in horses with recurrent airway obstruction

The goal of this study was to compare the rheological properties of mucus from horses with recurrent airway obstruction (RAO) to that from healthy controls during environmental challenge by stabling in stalls with straw as bedding and hay as feed. We determined viscoelasticity (log G* dyn/cm2, at 10 radian/s) and calculated mucociliary clearability index (MCI) and cough clearability index (CCI), which are derivative parameters of G* and the ratio of viscosity and elasticity measured at 1 and 100 radian/s, respectively. We also investigated the solids content of mucus, and cytology of bronchoalveolar lavage fluid (BALF). Samples were obtained before (0 h) and 6, 24 and 48 h after environmental challenge. The central findings were rheological changes in airway mucus, which occurred over time in RAO-affected animals, but not in controls. Mucus rheology was similar in both groups at 0 and 6 h. In RAO-affected horses, mucus viscoelasticity, as measured by log G*, increased from 2.49 +/- 0.18 dyn/cm2 (mean +/- s.e.) at 0 h to 3.05 +/- 0.13 dyn/cm2 at 24 h after environmental challenge, and was accompanied by significant decreases in MCI and CCI. Percent solids of mucus did not differ significantly between the 2 groups, nor over time. Rheological values did not correlate with BALF cytology. We conclude that viscoelastic properties of tracheal mucus samples from RAO horses in remission do not differ from those of normal horses. However, environmental challenge causes clinical signs of small airway disease and a concurrent increase in mucus viscoelasticity only in RAO horses. Therefore, we infer that unfavourable changes in mucus rheology may contribute to stasis and accumulation of mucus in RAO horses in exacerbation, but not in clinical remission.     

Pathophysiological changes occurring during Escherichia coli endotoxin and Pasteurella multocida challenge in piglets: relationship with cough and temperature and predicitive value for intensity of lesions

The aims of this study were (1) to correlate cough and body temperature (BT) with the severity of bronchopneumonia in pigs, (2) to determine whether these clinical signs can be used to early diagnose bronchopneumonia and (3) to assess the predictive values of cough and BT regarding lung lesions. Bronchopneumonia was induced by administering E. coli endotoxin (LPS) combined with Pasteurella multocida type A (PmA) in the trachea of 13 piglets. Saline-instilled negative controls (n = 8), PmA inoculated (n = 6) and LPS instilled (n = 5) groups were also constituted. Cough and BT were recorded daily while the bronchopneumonia severity was assessed using bronchoalveolar lavage fluid (BALF) cytology, cytokines and measurement of lung lesion volume. Changes in expiratory breathing pattern were also measured (Penh). The combination of LPS and PmA induced a subacute bronchopneumonia characterised by macrophage, neutrophil, and lymphocyte infiltration, changes in Penh and an increase in the mRNA level of IFN-gamma while IL8, IL-18 and TNF-alpha mRNA levels remained unchanged. The daily body weight gain of infected animals was significantly reduced. Cough and BT changes were proportional to the intensity of the lung inflammatory process, functional respiratory changes and to the extent of macroscopic lesions. When comparing the individual values of cough and BT to thresholds defined for both parameters, an early diagnosis of pneumonia was possible. Considering the pooled data of each group, it was possible to define thresholds allowing an early segregation between the groups of diseased and healthy piglets. The daily values of cough and BT were predictive for the volume of lung lesions recorded at the end of the trial. In conclusion, cough and BT appear as potential indicators for the intensity and the evolution of the respiratory disease. They also seem to be good predictors for the magnitude of lung lesions and weight gain recorded at the study endpoint.     

Effect of clenbuterol on histamine-induced airway obstruction in ponies

We determined the effect of IV administered beta 2-adrenergic receptor agonist clenbuterol on pulmonary function and on the response to histamine in 12 healthy ponies. Measurements were made at base line and after saline solution or clenbuterol was administered IV at a dosage of 0.2, 0.8, or 1.6 micrograms/kg. The dosage of clenbuterol used in each study was unknown to the investigators until all the data had been analyzed. Intravenous administration of saline solution or clenbuterol did not alter base-line pulmonary function significantly. Aerosol histamine administration significantly decreased arterial oxygen tension and dynamic compliance and increased pulmonary resistance, respiratory frequency, and minute ventilation, but had no effect on arterial carbon dioxide tension and tidal volume. The magnitude of change in these variables was unaffected by previous administration of clenbuterol at any of the dosages tested. We conclude that clenbuterol at the dosages tested is not a bronchodilator in healthy ponies and does not exert a significant protective effect against aerosol histamine-induced airway narrowing.     

Comparison of serum and urinary concentrations of clenbuterol with and without concomitant administration of furosemide in horses

Furosemide is frequently used to control or prevent exercise-induced pulmonary hemorrhage in performance horses. The bronchodilating agent clenbuterol is also commonly used as a treatment for inflammatory airway disease in performance horses. Use of both medications is regulated by many racing authorities. The effects of concomitant administration of furosemide and clenbuterol on the pharmacokinetics of clenbuterol have not been well characterized. A study was designed to evaluate the influence of furosemide on serum and urine concentrations of clenbuterol after oral administration of clenbuterol and intravenous administration of furosemide in horses. Results indicated that urinary concentrations of clenbuterol in horses treated concomitantly with furosemide and clenbuterol were increased, whereas serum concentrations of the drug were decreased. These effects persisted during the study period and varied among horses.     

Effects of obesity on lung function and airway reactivity in healthy dogs

The present study investigated the effects of bodyweight (BW) gain on respiratory function and airway responsiveness in healthy Beagles using barometric whole body plethysmography (BWBP). Six adult dogs were examined before and after a fattening diet. The high-energy diet induced a mean increase in BW of 41±6%. BWBP basal parameters were recorded prior to airway reactivity testing (using increasing concentrations of histamine nebulisations). An airway responsiveness index (H-Penh300) was calculated as the histamine concentration necessary to reach 300% of basal enhanced pause (Penh, bronchoconstriction index). The same dogs underwent a doxapram hydrochloride (Dxp) stimulation testing 2 weeks later. Basal measurements showed that obese dogs had tidal volume per kg (TV/BW) that was significantly decreased whilst respiratory rate (RR) increased significantly. H-Penh300 decreased significantly in obese Beagles, indicating increased bronchoreactivity. Dxp administration induced a significant increase in TV/BW, minute volume per kg (MV/BW), peak inspiratory and expiratory flows per kg (PIF/BW and PEF/BW) in both normal and obese dogs although the TV/BW increase was significantly less marked in the obese group. In conclusion, obesity induced changes in basal respiratory parameters, increased bronchoreactivity and a blunted response to Dxp-induced respiratory stimulation. This combination of basal respiratory parameters, bronchoreactivity testing and pharmacological stimulation testing using non-invasive BWBP can help characterize pulmonary function and airway responsiveness in obese dogs.     

A novel odor stimulation system in freely moving,behaving animals

Precise and reliable presentation of odorants to animals is crucial for olfactory studies. Although odor stimulation systems in anesthetized or awake, head-fixed animals are well established, temporally precise odor presentation in awake, freely moving animals remains a challenge. Here, we describe a new odor stimulation system which presents odors directly to the nostrils of freely moving mice. The system comprises 3 modules: an odor-delivery module, an odor-generation module, and a control module. The new system is precise and temporally reliable, and odor stimulation can be triggered by specific sniffing phases or other events. Moreover, the system can be combined with neural recordings, such as electrophysiology, and olfactory behavioral tests to investigate how neurons in the brain represent odor information during individual olfactory behaviors. This innovative odor stimulation system may replace traditional stimulation systems: It will enable precise odor presentation in a wide range of olfactory studies in freely moving animals.     

Evaluation of nebulised hay dust suspensions (HDS) for the diagnosis and investigation of heaves. 3: Effect of fractionation of HDS

To investigate the relative importance of inhaled particulates and soluble components in the response to inhaled hay dust suspension (HDS), heaves (previously termed chronic obstructive pulmonary disease; n = 7) and control (n = 6) horses were given inhalation challenges with whole and fractionated HDS. Fractionation was achieved by centrifugation to yield supernatant (SUP) and particulate debris. The particulates were then washed repeatedly in saline to produce a washed particulate (WP) fraction which comprised mainly fungal spores, and a wash fraction (WF) which comprised saline and soluble components washed from the surface of the particulates. Inhalation of HDS and SUP induced a significant airway neutrophilia in both groups, with the magnitude of the response being significantly greater in heaves horses. SUP induced significantly less airway neutrophilia than HDS in both groups, despite the endotoxin and protease content of HDS and SUP being comparable. WP and WF induced only a slight airway neutrophilia in heaves horses. However, a combined challenge with SUP and WP induced a neutrophilic response approaching the magnitude of that following HDS challenge, indicating that dust particulates contribute to the pulmonary recruitment of neutrophils in heaves. Consequently, inhalation challenge with HDS, which contains both particulates and soluble dust components, may be a more useful tool for the diagnosis and investigation of heaves than aqueous dust extracts, which contain only soluble components.     

Effects of aerosolized histamine and carbachol in the conscious horse.

Pulmonary function tests were performed in seven conscious, standing horses. Changes in pulmonary mechanics and ventilation volumes were measured after inhalation challenge with saline (baseline), histamine (1% w/v solution for 5 min) and carbachol (0.5% w/v solution for 3 min). Comparisons between baseline and posthistamine values revealed a significant (P less than 0.05) increase in nonelastic work of breathing (Wb), maximum change in transpulmonary pressure (max delta Ppl), and pulmonary resistance (RL), while dynamic compliance (Cdyn) decreased (P less than 0.05). Tripelennamine completely abolished these histamine induced changes suggesting the involvement of H1 receptors. A nonsignificant increase occurred in functional residual capacity. However, the amount of nitrogen retained in the lung at the end of a nitrogen washout test was significantly (P less than 0.05) greater after histamine when compared to baseline values. The effect of carbachol was qualitatively similar to that of histamine, Wb and max delta Ppl increased while Cdyn decreased (P less than 0.05). The increase in lower RL reached statistical significance (P less than 0.05) only at the beginning of expiration (/ 25% VT). The present investigation demonstrates that the physiological measurements of lung function could be carried out in conscious, unsedated horses and that the pulmonary function test methods could be used as a tool for study of drug induced changes in pulmonary mechanics.     

Airway hyperresponsiveness to adenosine 5’-monophosphate in feline chronic inflammatory lower airway disease

Airway hyperresponsiveness is a key feature of human asthma and chronic bronchitis and response to the indirectly acting agonist adenosine 5′-monophosphate (AMP) is thought to reflect underlying airway inflammation. To examine whether airway responsiveness testing (ART) with AMP may be used to differentiate healthy cats from those with asthma (FA) and chronic bronchitis (CB), 24 cats (9 FA, 6 CB, 9 controls) underwent ART with AMP at concentrations of 0.1, 1, 10, 100 and 500 mg/mL using barometric whole body plethysmography.The defined endpoint of ART, an increase in enhanced pause (Penh) exceeding 300% of the post-saline value (baseline), was reached in 9/15 patients (7 FA, 2 CB), but in none of the controls. Mean Penh (±SD) at baseline (BL) was 0.49 ± 0.16 for cases, and 0.54 ± 0.16 for controls, and was significantly increased after AMP challenge in clinical cases (2.62 ± 2.20), but not in controls (0.63 ± 0.30, P < 0.05). After separating responder (R) and non-responder (NR) cases, a more pronounced difference after challenge was found (R: 3.96 ± 1.84, NR: 0.6 ± 0.21, P < 0.001). The provocative concentration of the agonist that increased Penh to 300% of BL (PC Penh 300) in R cases was 52.98 ± 48.04 mg/mL AMP. Age had no influence on the responder status or PC Penh 300. It was concluded that AMP challenge may offer a new method for the identification of cats with lower inflammatory airway disease, and possibly for monitoring disease progression or response to therapy.     

The effects of clenbuterol hydrochloride on the pulmonary function of calves with subacute and chronic bronchopneumonia

The influence of clenbuterol on some pulmonary function tests in 11 calves with subacute and chronic bronchopneumonia has been investigated. Treatment with clenbuterol caused changes only in calves with subacute bronchopneumonia; in these animals it decreased the total pulmonary resistance, the viscous work per litre and the peak to peak changes in transpulmonary pressure.     

Comparison of bronchodilators in alleviating clinical signs in horses with chronic obstructive pulmonary disease

A representative of each of 3 types of bronchodilators was given iv to horses with clinical signs of chronic obstructive pulmonary disease. We used atropine (0.01 mg/kg of body weight) as an anticholinergic, isoproterenol HCl as a beta-adrenergic, and aminophylline, the soluble salt of theophylline, as a methylxanthine. Response to these drugs was evaluated by measuring maximal change in intrathoracic pressures (delta Ppl) and observing clinical signs. Atropine caused a significant (P less than 0.01) reduction in delta Ppl, compared with that caused by aminophylline. Atropine caused the delta Ppl to decrease an average of 83 +/- 15% toward normal. Clinical improvement was noticed in 87% of the cases. Maximal changes in pleural pressures were reduced by an average of 21 +/- 14 mm of Hg, with 27% returning completely to normal. Isoproterenol helped 77% of horses clinically, and caused delta Ppl to decrease an average of 68 +/- 31% toward normal. The average reduction in delta Ppl was 16 +/- 10 mm of Hg. Aminophylline helped alleviate clinical signs of 50% of the horses and caused delta Ppl to decrease an average of 41 +/- 36% toward normal. The average reduction in delta Ppl was 10 +/- 11 mm of Hg.     

Cardiopulmonary effects of clenbuterol in the horse

Clenbuterol, a bronchospasmolytic agent (β₂ agonist) was studied in terms of its hemodynamic and airflow response in eight, healthy horses. Four animals were instrumented to record intrapleural pressure and air flow, these were used to compute pulmonary resistance, peak flow rates, and tidal volumes. Four animals were instrumented to record pulmonary arterial pressure, carotid arterial pressure, cardiac output, and arterial gas tensions. After control values were recorded, clenbuterol (0.8 μg/kg) was intravenously administered to each horse in each experiment group.
Following clenbuterol administration, non-elastic resistance of the lung or pulmonary resistance significantly decreased, 33.6% reduction at 10 min post-clenbuterol. Pulmonary resistance remained lowered during the entire procedure and showed no tendency of returning toward control values by 3 h post-clenbuterol. Within 30 sec following clenbuterol injection carotid arterial pressure decreased (mean pressure decrease 28.2%). Accompanying the change in arterial pressure, the heart rate drastically increased, 99.0%. Both changes were transient and returned to control ranges within 2 min.
Clenbuterol appears to be effective in reducing non-elastic resistance of the lung, however intravenous administration to an animal with pre-existing cardiovascular or cardiopulmonary disease should be avoided.