A multidisciplinary approach to reduce and refine antimicrobial drugs use for diarrhoea in dairy calves.

This study evaluated the impact of a multidisciplinary intervention to reduce and refine antimicrobial drug use for treatment of diarrhoea in dairy calves. The intervention consisted of modification of management practices for disease prevention, health evaluation training of farm staff, implementation of an algorithm directed at antimicrobial therapy for diarrhoeic calves, and monthly farm visits. A combination of retrospective (before intervention period [BP], 1 year) and prospective (immediately after intervention period [AP], 1 year) cohorts were used. Health outcomes measured included incidence of diarrhoea and overall mortality. Antimicrobial treatment rates for diarrhoea and total mass of antimicrobial drugs used at the calf and farm level were also evaluated. Outcomes were assessed using the χ2 or Fisher's exact test, and odds ratios and 95% confidence intervals (95% CI) were calculated. Records of 2049 and 2251 calves from 10 farms were available in the BP and AP, respectively. Reduced antimicrobial treatment rates occurred on seven farms (P < 0.01), but not on three farms in the AP. A total of 85% (1303/1573) and 18% (310/1698) of diarrhoeic calves were treated with antimicrobial drugs in the BP and AP (P < 0.001), respectively. There were no differences in the incidence of diarrhoea or mortality between periods. In the AP, the use sulfamethazine and trimethoprim decreased on eight farms, one farm discontinued use of lincomycin and spectinomycin, while two farms discontinued use and one reduced use of cephalosporins. This multidisciplinary approach was effective in reducing antimicrobial drug use for calf diarrhoea on dairy farms without negative impacts on calf health.

Detection of Bovine Coronavirus in Healthy and Diarrheic Dairy Calves.

BACKGROUND: BCoV is identified in both healthy and diarrheic calves, complicating its assessment as a primary pathogen. OBJECTIVES: To investigate the detection rates of bovine coronavirus (BCoV) in feces of healthy and diarrheic calves and to describe the usefulness of a pancoronavirus reverse transcriptase (RT) PCR (PanCoV-RT-PCR) assay to identify BCoV in samples of diarrheic calves. ANIMALS: Two hundred and eighty-six calves <21 days. Calves with liquid or semiliquid feces, temperature >39.5°C, and inappetence were considered as cases, and those that had pasty or firm feces and normal physical examination were designated as controls. METHODS: Prospective case-control study. A specific BCoV-RT-PCR assay was used to detect BCoV in fecal samples. Association between BCoV and health status was evaluated by exact and random effect logistic regression. Fecal (n = 28) and nasal (n = 8) samples from diarrheic calves were tested for the presence of BCoV by both the PanCoV-RT-PCR and a specific BCoV-RT-PCR assays. A Kappa coefficient test was used to assess the level of agreement of both assays. RESULTS: BCoV was detected in 55% (157/286) of calves; 46% (66/143), and 64% (91/143) of healthy and diarrheic calves, respectively. Diarrheic calves had higher odds of BCoV presence than healthy calves (OR: 2.16, 95% CI: 1.26 to 3.83, P = 0.004). A good agreement between PanCoV-RT-PCR and BCoV-RT-PCR to detect BCoV was identified (κ = 0.68, 95% CI: 0.392 to 0.967; P < 0.001). CONCLUSIONS AND CLINICAL IMPORTANCE: BCoV was more likely to be detected in diarrheic than healthy calves. The PanCoV-RT-PCR assay can be a useful tool to detect CoV samples from diarrheic calves.

Characterization of the Fecal Bacterial Microbiota of Healthy and Diarrheic Dairy Calves.

BACKGROUND: Neonatal diarrhea accounts for more than 50% of total deaths in dairy calves. Few population-based studies of cattle have investigated how the microbiota is impacted during diarrhea. OBJECTIVES: To characterize the fecal microbiota and predict the functional potential of the microbial communities in healthy and diarrheic calves. METHODS: Fifteen diarrheic calves between the ages of 1 and 30 days and 15 age-matched healthy control calves were enrolled from 2 dairy farms. The Illumina MiSeq sequencer was used for high-throughput sequencing of the V4 region of the 16S rRNA gene (Illumina, San Diego, CA). RESULTS: Significant differences in community membership and structure were identified among healthy calves from different farms. Differences in community membership and structure also were identified between healthy and diarrheic calves within each farm. Based on linear discriminant analysis effect size (LEfSe), the genera Bifidobacterium, Megamonas, and a genus of the family Bifidobacteriaceae were associated with health at farm 1, whereas Lachnospiraceae incertae sedis, Dietzia and an unclassified genus of the family Veillonellaceae were significantly associated with health at farm 2. The Phylogenetic Investigation of Communities Reconstruction of Unobserved States (PICRUSt) analysis indicated that diarrheic calves had decreased abundances of genes responsible for metabolism of various vitamins, amino acids, and carbohydrate. CLINICAL RELEVANCE: The fecal microbiota of healthy dairy calves appeared to be farm specific as were the changes observed during diarrhea. The differences in microbiota structure and membership between healthy and diarrheic calves suggest that dysbiosis can occur in diarrheic calves and it is associated with changes in predictive metagenomic function.

Comparison of manual and suction pump aspiration techniques for performing bronchoalveolar lavage in 18 dogs with respiratory tract disease.

BACKGROUND: Different aspiration techniques to retrieve bronchoalveolar lavage fluid (BALF) affect sample quality in healthy dogs. Studies evaluating these techniques in dogs with respiratory disease are lacking. OBJECTIVES: To compare sample quality of BALF acquired by manual aspiration (MA) and suction pump aspiration (SPA). ANIMALS: Eighteen client-owned dogs with respiratory disease. METHODS: Randomized, blinded prospective clinical trial. Manual aspiration was performed with a 35-mL syringe attached directly to the bronchoscope biopsy channel and SPA was performed with a maximum of 50 mmHg negative pressure applied to the bronchoscope suction valve using the suction trap connection. Both aspiration techniques were performed in each dog on contralateral lung lobes, utilizing 2 mL/kg lavage volumes per site. Samples of BALF were analyzed by percentage of retrieved infusate, total nucleated cell count (TNCC), differential cell count, semiquantitative assessment of slide quality, and diagnosis score. Data were compared by paired Student's t-test, Wilcoxon signed-rank test, chi-squared test, and ANOVA. Cohen's kappa coefficient was used to assess agreement. RESULTS: The percentage of retrieved BALF (P = .001) was significantly higher for SPA than MA. Substantial agreement was found between cytologic classification of BALF obtained with MA and SPA (kappa = 0.615). There was no significant difference in rate of definitive diagnosis achieved with cytologic assessment between techniques (P = .78). CONCLUSIONS AND CLINICAL IMPORTANCE: Suction pump aspiration, compared to MA, improved BALF retrieval, but did not significantly affect the rate of diagnostic success of bronchoalveolar lavage (BAL) in dogs with pulmonary disease.

The use of a tongue tie alters laryngohyoid position in the standing horse.

REASONS FOR PERFORMING STUDY: The use of tongue ties in racing is common, yet there are few data to support their efficacy. In order to make appropriate recommendations for clinical practice and policy on tongue ties, data documenting their effect on upper airway structure are necessary. OBJECTIVES: To determine the effect of a tongue tie on the resting laryngohyoid position of the standing horse. STUDY DESIGN: Experimental study. METHODS: Twelve normal Standardbred horses were subjected to ultrasonographic measures of laryngohyoid position during 3 phases of tack application: Phase I--halter and lead; Phase II--bit, bridle, harness and check applied; and Phase III--as in Phase II but with the tongue tie added. RESULTS: Compared to Phase I, during Phase III with the application of a tongue tie a significant difference between lingual process position was noted both rostrally and caudally (P<0.001 and P<0.001), such that the tongue tie resulted in an increase in lingual process depth. The tongue tie in Phase III resulted in a decrease in depth of the thyroid cartilage and basihyoid bone compared to the Phase I location (P = 0.007 and P = 0.0047). CONCLUSION: The use of a tongue tie has a significant effect on the basihyoid and thyroid cartilage positions in the standing horse. POTENTIAL RELEVANCE: This is the first report documenting a difference in laryngohyoid morphology following the application of a tongue tie, providing evidence that the use of a tongue tie has a measurable effect on upper airway structure. The functional implications of this finding are yet to be elucidated.

Effect of frusemide on transvascular fluid fluxes across the lung in exercising horses.

REASONS FOR PERFORMING STUDY: Frusemide (Fru) is widely prescribed for management of racehorses experiencing EIPH. The effect of Fru in the lung appears to be a reduction in transcapillary pressures and inhibition of the erythrocyte anion exchange, which may lead to attenuation of transpulmonary fluid fluxes during exercise. HYPOTHESIS: Treatment with Fru will attenuate transpulmonary fluid fluxes in horses during high intensity exercise. METHODS: In a crossover study, 6 race-fit Standardbred horses were treated with 250 mg of Fru i.v. (FruTr) or placebo (Con) 4 h before exercise on a high speed treadmill until fatigue. Arterial and central mixed venous blood, as well as CO(2) elimination and O(2) uptake, were sampled. Volume changes across the lung and transvascular fluid fluxes were calculated from changes in haemoglobin, packed cell volume, plasma protein and cardiac output (Q). RESULTS: During exercise, Q increased in both Con and FruTr, with Q being significantly lower in FruTr (mean ± s.e. 301.8 ± 8.5 l/min at fatigue) compared to Con (336.5 ± 15.6 l/min) (P<0.01). At rest frusemide had no effect on erythrocyte (J(ER)) and transvascular (J(V-A)) fluid fluxes across the lung. Exercise had a significant effect on J(ER) and J(V-A) (P ≤ 0.02). During exercise, J(ER) (at fatigue 14.6 ± 2.3 l/min and 11.6 ± 2.2 l/min in Con and FruTr, respectively) and J(V-A) (at fatigue 14.9 ± 2.3 l/min and 12.0 ± 2.2 l/min in Con and FruTr, respectively) were not significantly different between Con and FruTr (P = 0.6 and P = 0.8 for J(ER) and J(V-A), respectively). CONCLUSIONS AND CLINICAL IMPORTANCE: Fru does not have a measurable effect on J(ER) and J(V-A). Cardiac output was reduced in FruTr, suggesting that there were also smaller changes in the capillary recruitment and transvascular transmural hydrostatic pressures; however, this did not effect J(V-A). Therefore, Fru at the dose of 250 mg does not appear to be an effective treatment for regulating pulmonary transvascular forces during exercise in horses.

Effects of chronic acetazolamide administration on gas exchange and acid-base control in pulmonary circulation in exercising horses.

REASONS FOR PERFORMING STUDY: Carbonic anhydrase (CA) catalyses the hydration/dehydration reaction of CO(2) and increases the rate of Cl(-) and HCO(3)(-) exchange between the erythrocytes and plasma. Therefore, chronic inhibition of CA has a potential to attenuate CO(2) output and induce greater metabolic and respiratory acidosis in exercising horses. OBJECTIVES: To determine the effects of Carbonic anhydrase inhibition on CO(2) output and ionic exchange between erythrocytes and plasma and their influence on acid-base balance in the pulmonary circulation (across the lung) in exercising horses with and without CA inhibition. METHODS: Six horses were exercised to exhaustion on a treadmill without (Con) and with CA inhibition (AczTr). CA inhibition was achieved with administration of acetazolamide (10 mg/kg bwt t.i.d. for 3 days and 30 mg/kg bwt before exercise). Arterial, mixed venous blood and CO(2) output were sampled at rest and during exercise. An integrated physicochemical systems approach was used to describe acid base changes. RESULTS: AczTr decreased the duration of exercise by 45% (P < 0.0001). During the transition from rest to exercise CO(2) output was lower in AczTr (P < 0.0001). Arterial PCO(2) (P < 0.0001; mean ± s.e. 71 ± 2 mmHg AczTr, 46 ± 2 mmHg Con) was higher, whereas hydrogen ion (P = 0.01; 12.8 ± 0.6 nEq/l AczTr, 15.5 ± 0.6 nEq/l Con) and bicarbonate (P = 0.007; 5.5 ± 0.7 mEq/l AczTr, 10.1 ± 1.3 mEq/l Con) differences across the lung were lower in AczTr compared to Con. No difference was observed in weak electrolytes across the lung. Strong ion difference across the lung was lower in AczTr (P = 0.0003; 4.9 ± 0.8 mEq AczTr, 7.5 ± 1.2 mEq Con), which was affected by strong ion changes across the lung with exception of lactate. CONCLUSIONS: CO(2) and chloride changes in erythrocytes across the lung seem to be the major contributors to acid-base and ions balance in pulmonary circulation in exercising horses.

Immunohistochemical identification of collagen in the equine lung.

Recurrent airway obstruction (RAO) in the horse is a disease characterized by reversible bronchoconstriction and by mucus and neutrophil accumulation in the airways. It has been hypothesized that in horses with RAO, remodeling changes occur that are similar to those described in humans with asthma. Although collagen fibrils are present surrounding normal airways, they are a prominent feature of airway remodeling in human asthma with evidence of enhanced collagen III and I fibril deposition. An immunolabeling method was developed to identify collagen I and III in equine lung and to describe the collagen fiber type and distribution within the walls of the noncartilagenous bronchioles. The health status of 14 horses was characterized by clinical respiratory exam, bronchoalveolar lavage cytology, and pulmonary function tests. Following postmortem examination and histological assessment, horses were divided into RAO-affected (n = 4) and nonaffected (n = 10) groups. Eight sections per horse from all lung regions were evaluated histologically. Results of the study showed that collagens I and III were present in the lamina propria and adventitial area of the noncartilaginous bronchioles. There was clear staining differentiation between collagen I or III, airway smooth muscle, and the airway epithelium. Collagen I and III were present in the lamina propria and adventitial areas of the noncartilaginous bronchioles of horses, and there was no significant difference in the relative amount of collagen I and III between this group of RAO-affected and nonaffected horses.

Finite element analysis of wall stress in the equine pulmonary artery.

REASONS FOR PERFORMING STUDY: Arterial calcification is found frequently in the pulmonary artery of racehorses, but the aetiology is unknown. Calcification might be associated with increased wall stress due to arterial geometry (shape) and exercise-induced hypertension. HYPOTHESIS: High wall stress levels are found in the regions associated with calcified lesion formation, exacerbated as transluminal pressure increases to levels associated with exercise. METHODS: The pulmonary arteries of 5 horses, unaffected by calcification, were dissected and pressurised to resting and exercising physiological transluminal pressures and scanned with MRI. Arterial geometries were reconstructed to form 3D computer models and finite element analyses performed. Wall stress levels were measured in 4 regions of interest: the arterial trunk and bifurcation, the wall ipsilateral and contralateral to the bifurcation. Measurements were made for arterial transluminal pressures of 25, 50 and 100 mmHg. RESULTS: High wall stress levels were consistently found at the pulmonary artery bifurcation and wall ipsilateral to the bifurcation, where calcified lesions typically form. Lower wall stress levels were found along the trunk and the wall contralateral to the bifurcation where lesions are less frequently found. Wall stress levels increased 5-fold over a 4-fold increase in pressure. The wall stress levels ranged 10 kPa in the wall of the branch contralateral to the bifurcation at 25 mmHg to 400 kPa in the bifurcation at 100 mmHg. CONCLUSIONS: Wall stress from arterial geometry and increased pulmonary artery transluminal pressure are factors that may be associated with calcification of the equine pulmonary artery. POTENTIAL RELEVANCE: Arterial calcification may increase the risk of arterial wall failure in racing horses.

Clara cell secretory protein is reduced in equine recurrent airway obstruction.

Horses are prone to recurrent airway obstruction (RAO), an inflammatory lung disease induced by repeated exposure to environmental mold, dust, and bacterial components. Active disease manifests with mucus hyperproduction, neutrophilic inflammation, bronchoconstriction, and coughing. Chronically affected animals have lung remodeling characterized by smooth muscle hyperplasia, collagen deposition, lymphoid hyperplasia, and impaired aerobic performance. Clara cell secretory protein (CCSP) counters inflammation in the lung, hence we hypothesized that CCSP depletion is a key feature of RAO in horses. Recombinant equine CCSP and specific antiserum were produced, and percutaneous lung biopsies were obtained from 3 healthy horses and from 3 RAO-affected horses before and after induction of RAO. CCSP relative gene expression in tissue, as well as protein concentration in lung lavage fluid, was determined. Immunocytochemical analysis, using both light and immunogold ultrastructural methods, demonstrated reduced CCSP staining in lung tissue of animals with RAO. Immunogold label in Clara cell granules was less in animals with chronic RAO than in normal animals, and absent in animals that had active disease. Median lung lavage CCSP concentration was 132 and 129 ng/ml in healthy horses, and 62 and 24 ng/ml in RAO horses before and after challenge, respectively. CCSP lung gene expression was significantly higher in healthy animals than in animals with chronic RAO. Together, these preliminary findings suggest that reduced production of CCSP and subcellular changes in Clara cells are features of chronic environmentally induced lung inflammation in horses.

Histamine bronchoprovocation does not affect bronchoalveolar lavage fluid cytology, gene expression and protein concentrations of IL-4, IL-8 and IFN-gamma.

In diagnosing inflammatory airway disease (IAD) in performance horses, a histamine bronchoprovocation (HBP) test is often performed. In previously published studies, HBP is usually undertaken prior to cytological examination of the bronchoalveolar lavage (BAL) cells. The purpose of this study was to determine if HBP alters (1) the total nucleated cell numbers and distribution in BAL fluid (BALF) and (2) the mRNA and protein concentrations of selected cytokines in BAL cells and BALF, respectively. BALF was initially collected endoscopically from the right middle or diaphragmatic lung lobe in eight healthy young Standardbred horses. Five to six days later, HBP was performed by aerosolization of histamine (8mg) over a 2min period. BALF was again collected within 2-4h of the HBP from the left middle or diaphragmatic lung lobe. In both samples, total and differential WBC counts were obtained. The gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR. The cytokine protein concentrations were measured in the BALF using ELISA. HBP was not associated with either a change in the total BAL cell number or in the distribution of the BAL cells. BAL cell expression of IL-4, IL-8 and IFN-gamma, detected in all samples with the exception of IL-4 in one horse (post-HBP), was not altered as a result of HBP. HBP was not associated with a significant change in IL-8 or IFN-gamma concentrations in the BALF. IL-4 protein was undetectable in BALF either prior to or following HBP. We conclude that HBP can precede BALF collection performed within 2-4h of the former without affecting selected parameters analysed in the BAL cells or BALF.

Arterial calcification in race horses.

Calcification of large arteries has been sporadically reported in horses. The pathogenesis is still unknown, but recent studies in humans suggest that this is a regulated biomineralizing process. This study surveyed the prevalence, distribution, and severity of vascular calcification in Thoroughbred and Standardbred racehorses. Histopathologic, ultrastructural imaging, and energy dispersive X-ray elemental analyses were used to examine the lesions. Calcification of the tunica media, predominantly the pulmonary artery, was found in 82% of horses (83/101). Young adult horses (mean [SD] age in years, 4.44 +/- 2.17) of both breeds and sexes were similarly affected. Lesions appeared as white-to-yellowish, hard, and gritty plaques of variable size. On microscopic examination, elastic fibers within the tunica media were thinned, fragmented, and calcified, and surrounded by dense collagen matrix. Elemental analysis showed distinct peaks for calcium and phosphorus, consistent with hydroxyapatite mineral. The frequent occurrence of calcification in the tunica media of large pulmonary arteries of young racing horses indicates the need to investigate its pathogenesis and potential clinical implications.

Effects of chronic acetazolamide administration on fluid flux from the pulmonary vasculature at rest and during exercise in horses.

REASONS FOR PERFORMING STUDY: Horses develop high pulmonary pressures during exercise, which force fluid out of pulmonary capillaries. Specific airway diseases in horses, especially those associated with hypoxaemia, hypercapnoea and acidosis may influence pulmonary haemodynamics and pulmonary interstitial fluid equilibrium. OBJECTIVES: This study was designed to determine fluid flux (J(V-A) l/min) across the lung in exercising horses treated chronically with acetazolamide. METHODS: Six horses were exercised on a treadmill until fatigue without (Con) and with chronic carbonic anhydrase (CA) inhibition (AczTr) and associated hypercapnoea and acidosis. Carbonic anhydrase inhibition was achieved with administration of acetazolamide (Acz). Arterial and mixed venous blood were sampled, and VCO2 and VO2 measured. Blood volume changes across the lung (deltaBV%) were calculated from changes in plasma protein, haemoglobin and packed cell volume (PCV). Cardiac output (Q) was calculated using Fick principle. J(V-A) across the alveolar-capillary barrier was then quantified based on Q and deltaBV. Variables were analysed using 2-way repeated-measures ANOVA (P<0.05). A significant F ratio was further analysed using Tukey post hoc analysis. RESULTS: Treatment had a significant effect on J(V-A) (P = 0.002). At rest there was no J(V-A) in Con (0.63 +/- 0.6 l/min) and AczTr (0.84 +/- 0.3 l/min). During exercise Con fluid moved from the pulmonary circulation into the pulmonary interstitium (mean +/- s.e. J(V-A) 9.4 +/- 2.4 l/min). This was different from AczTr (mean +/- s.e. J(V-A) 1.8 +/- 1.9 l/min), where no transvascular fluxes from pulmonary circulation were present during exercise (P = 0.008). CONCLUSIONS: Chronic Acz treatment with associated hypercapnoea and acidosis affects J(V-A) in lungs of exercising horses. Lung fluid dynamics adapted to hypercapnoea and acidosis with reduction of fluid flow from the pulmonary circulation. POTENTIAL RELEVANCE: The current data provide comprehensive evidence of in vivo fluid homeostasis in lungs of exercising horses without and with CA inhibition.

Effects of pulmonary abscesses on racing performance of horses treated at referral veterinary medical teaching hospitals: 45 cases (1985-1997).

OBJECTIVE: To determine whether results of physical or radiographic examination or biochemical analyses in adult racehorses with primary lung abscesses were associated with ability to race following treatment. DESIGN: Multiple-center retrospective study. ANIMALS: 25 Standardbreds and 20 Thoroughbreds. PROCEDURE: Medical records of horses with a primary lung abscess that were admitted to any of 4 veterinary teaching hospitals were reviewed. Results of physical examination, laboratory testing, and thoracic radiography were reviewed. Racing performance after treatment was compared with performance before illness and with performance of the general population of racehorses of similar age, sex, and breed. RESULTS: 23 of 25 Standardbreds and 13 of 20 Thoroughbreds raced after diagnosis and treatment of a lung abscess. Most horses had a solitary abscess in the dorsal to caudodorsal lung fields. Results of initial physical examination, biochemical analyses, and culture and identification of the microbial isolate were not associated with whether a horse returned to racing. For horses that had raced prior to the illness, race performance after treatment of the lung abscess was not significantly different from performance before the illness. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of racing performance in those horses that resumed racing after treatment, long-term residual lung damage did not develop in horses with primary lung abscesses that were treated appropriately. It is not known whether horses that recovered would be more likely to bleed from the site of a prior infection when resuming strenuous exercise and whether lung abscesses contributed to a failure to resume racing.

In vitro allergen-induced degranulation of pulmonary mast cells from horses with recurrent airway obstruction (heaves).

OBJECTIVE: To determine the capacity of pulmonary mast cells (PMC) to degranulate in response to various potential allergens and other secretagogues in horses with recurrent airway obstruction (heaves) and clinically normal horses before and after exposure to moldy hay. ANIMALS: 5 horses with heaves and 5 clinically normal horses. PROCEDURES: Heaves was characterized as an increased clinical respiratory score and maximum change in transpulmonary pressure of > 20 cm H2O after exposure. Bronchoalveolar lavage was performed during each period. Washed and resuspended cells were exposed for 20 minutes at 37 C with whole reconstituted freeze-dried preparations of Aspergillus fumigatus, Alternaria tenuis, and Ambrosia elatior, fungal extracts of Aspergillus fumigatus, Alternaria tenuis, and Micropolyspora faeni; A23187; and compound 48/80. Histamine release (HR) was used as a marker of degranulation. RESULTS: Compared with clinically normal horses, HR was significantly greater from PMC from horses with heaves during remission and exacerbation in response to whole preparations and extracts of Aspergillus fumigatus and whole preparations of Alternaria tenuis. Extracts of Alternaria tenuis caused significantly greater HR from PMC from horses with heaves during exacerbation. Histamine was also released from PMC in response to A23187 and to changes in osmolality of the medium, but only as a result of cell lysis by compound 48/80. CONCLUSIONS: Increased degranulation of PMC after antigenic challenge may contribute to the pathogenesis of heaves in horses. CLINICAL RELEVANCE: Strategies for prevention and treatment that attenuate degranulation of PMC may assist in the clinical management of horses with heaves.

Bovine herpesvirus 1 (BHV1) seroprevalence in the breeding cattle population of the Veneto region: prospects for the implementation of a control programme.

The results of a serological survey for bovine herpes virus (BHV1) antibodies in the breeding cattle population of the Veneto region are presented. The data do not support the hypothesis of an high prevalence of BHV1; on farms where vaccination was not carried out most animals were seronegative, and seropositive animals were generally older. Therefore, when drawing up the guidelines for a control programme, systematic immunization (with glycoprotein E-deleted vaccines) should be restricted only to farms with a high prevalence of BHV1 antibodies and/or with a high risk of BHV1 occurrence; in most unvaccinated farms a 'test and removal' policy appears to be more appropriate in order to rapidly eradicate BHV1 from the entire stock.

Associations between physical examination, laboratory, and radiographic findings and outcome and subsequent racing performance of foals with Rhodococcus equi infection: 115 cases (1984-1992).

OBJECTIVE: To determine whether physical examination, laboratory, or radiographic abnormalities in foals with Rhodococcus equi infection were associated with survival, ability to race at least once after recovery, or, for foals that survived and went on to race, subsequent racing performance. DESIGN: Retrospective study. ANIMALS: 49 Thoroughbreds and 66 Standardbreds admitted to 1 of 6 veterinary teaching hospitals between 1984 and 1992 in which R equi infection was positively diagnosed. PROCEDURE: Results of physical examination, laboratory testing, and thoracic radiography were reviewed. Indices of racing performance were obtained for foals that recovered and eventually raced and compared with values for the US racing population. RESULTS: 83 (72%) foals survived. Foals that did not survive were more likely to have extreme tachycardia (heart rate > 100 beats/min), be in respiratory distress, and have severe radiographic abnormalities on thoracic radiographs at the time of initial examination than were foals that survived. Clinicopathologic abnormalities were not associated with whether foals did or did not survive. Forty-five of the 83 surviving foals (54%) eventually raced at least once, but none of the factors examined was associated with whether foals went on to race. Racing performance of foals that raced as adults was not significantly different from that of the US racing population. CLINICAL IMPLICATIONS: R equi infection in foals is associated with a decreased chance of racing as an adult; however, foals that eventually go on to race perform comparably to the US racing population.

Pulmonary eosinophilia associated with increased airway responsiveness in young racing horses.

Horses are known to acquire small airway disease (SAD), an allergen-induced naturally occurring syndrome of reversible obstructive lung disease accompanied by airway hyperresponsiveness and increased inflammatory cell numbers on bronchoalveolar lavage (BAL). This disorder has received scant attention in young racehorses. The purpose of the present report was to examine the effect of BAL eosinophilia in young racehorses on clinical examination, BAL, hematology, airway responsiveness, and on pulmonary function at rest and after a standardized exercise challenge. Five (3 males, 2 females; age 2.6 +/- 0.9 years) with a history of respiratory compromise and BAL eosinophil differential count > 5% and 6 controls (4 males, 2 females; age 3.5 +/- 1.0 years) training and performing to expectation with normal BAL cell differential (eosinophils < 1%) were studied. Respiratory system clinical examination was performed and expressed as a clinical score. Arterial blood gas measurements, CBC, and pulmonary function testing were performed at rest. Pulmonary mechanics measurements were repeated 1 hour and 20 hours after a standardized treadmill exercise challenge. Incremental histamine inhalation challenge was performed and the concentration of histamine effecting a 35% decrease in dynamic compliance (PC35CDyn) was determined. Significant differences were noted between and controls with regard to clinical score (P = .01), blood eosinophils (P = .04), BAL cell count (P = .04), BAL macrophage differential (P = .04), PC35CDyn (P = .008), and tidal volume and respiratory rate at 20 hours following exercise challenge (P = .05). We conclude that pulmonary eosinophilia and airway hyperresponsiveness are manifest in some young horses without overt airway obstruction at rest. We speculate that these may be early events in the natural progression of heaves.

Evaluation of an in vitro degranulation challenge procedure for equine pulmonary mast cells.

Pulmonary mast cells (PMC) are important components of the inflammatory process in equine allergic lung diseases such as heaves. Very little, however, is known of the degranulation kinetics of these cells and thus, their pathophysiologic role remains largely speculative. The purpose of this study was to develop a repeatable protocol for in vitro equine PMC degranulation. Five mature horses (sex: 2 M, 3 F; age: 8.8 +/- 6.5 y), historically free of pulmonary disease and normal on clinical respiratory examination, arterial blood gas analysis, pulmonary mechanics testing and histamine inhalation challenge, were studied. Bronchoalveolar lavage was performed on 4 separate occasions, at least 2 d apart, in a different lung lobe on each occasion. The lavage fluid was concentrated by centrifugation. Cells were resuspended in modified HEPES/Tyrode, assessed for viability by Trypan blue exclusion, and PMC concentration determined. Cell inocula containing 30,000 PMC were incubated with 10(-8) to 6 x 10(-5) M A23187. Cells were then separated by centrifugation and histamine release (HR) was determined by fluorometric assay. The procedure was readily performed and yielded sufficient PMC for 30 to 60 inocula per lavage. Maximal HR (34.4 +/- 16.1%) was obtained with 10(-5) M A23187. The degranulation process was largely complete by 20 min but cell lysis was negligible. The challenge was repeatable within horse and produced a mean coefficient of variability of 23.0% following 20 min incubation with 10(-5) M A23187. We conclude that equine PMC degranulation can be repeatably performed in vitro and speculate that this protocol may be useful in further studies on the pathophysiology and treatment of equine allergic lung diseases.

Pathogenesis and clinical signs of equine herpesvirus-1 in experimentally infected ponies in vivo.

Equine herpesvirus-1 (EHV-1) causes respiratory disease, neonatal death, abortion and neurologic disease. The main purpose of this study was to identify viral antigen in respiratory tract samples by immunoperoxidase staining. Six pony foals were selected on the basis of demonstrating seronegativity to EHV-1 by virus neutralization and housed in isolation. They were infected experimentally by administering EHV-1 nebulized ultrasonically through a face mask. Successful infection was clinically apparent as each of the foals had febrile responses, nasal discharge, and enlarged submandibular lymph nodes. Sporadic coughing was also heard. EHV-1 was isolated from nasopharyngeal swabs of 4/6 ponies and seroconversion was demonstrated in all foals. Bronchoscopic examination of the large airways revealed hyperemia. The incidence of recovery of Actinobacillus suis from nasopharyngeal swabs increased initially, with recovery of Streptococcus zooepidemicus isolates predominating at 3 wk post-infection. Cytology brushes were used to sequentially sample the respiratory tract of the infected ponies at the nasopharynx, mid-trachea and the mainstem bronchus. Bronchoalveolar lavage provided lung cells. Immunocytochemistry techniques were applied to both types of samples to locate EHV-1 antigen. Indirect immunoperoxidase staining of samples utilizing monoclonal antibodies specific for EHV-1 demonstrated viral antigen associated with cellular debris, primarily in the nasopharyngeal samples on days 3-9 post-infection.