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There is a growing trend of animal-assisted intervention (AAI) programs on college campuses to improve mental health among college students through human–animal interactions. The purpose of this pr...

Prison-equine programs (PEPs) appear to play a role in rehabilitating incarcerated persons (IPs) by teaching equine and farrier skills, as well as promoting therapeutic effects through the human-an...

OBJECTIVE To evaluate the lipidomic profile of surfactant obtained from horses with asthma at various clinical stages and to compare results with findings for healthy horses exposed to the same conditions. SAMPLE Surfactant samples obtained from 6 horses with severe asthma and 7 healthy horses. PROCEDURES Clinical evaluation of horses and surfactant analysis were performed. Samples obtained from horses with severe asthma and healthy horses before (baseline), during, and after exposure to hay were analyzed. Crude surfactant pellets were dried prior to dissolution in a solution of isopropanol:methanol:chloroform (4:2:1) containing 7.5mM ammonium acetate. Shotgun lipidomics were performed by use of high-resolution data acquisition on an ion-trap mass spectrometer. Findings were analyzed by use of an ANOVA with a Tukey-Kramer post hoc test. RESULTS Results of lipidomic analysis were evaluated to detect significant differences between groups of horses and among exposure statuses within groups of horses. Significantly increased amounts of cyclic phosphatidic acid (cPA) and diacylglycerol (DAG) were detected in surfactant from severely asthmatic horses during exposure to hay, compared with baseline and postexposure concentrations. Concentrations of cPA and DAG did not change significantly in healthy horses regardless of exposure status. CONCLUSIONS AND CLINICAL RELEVANCE cPA 16:0 and DAG 36:2 were 2 novel lipid mediators identified in surfactant obtained from asthmatic horses with clinical disease. These molecules were likely biomarkers of sustained inflammation. Further studies are needed to evaluate a possible correlation with disease severity and potential alterations in the plasma lipidomic profile of horses with asthma.

BACKGROUND Equine protozoal myeloencephalitis (EPM) is a common and devastating neurologic disease of horses in the United States. Because some EPM-affected horses have decreased immune responses, immunomodulators such as levamisole have been proposed as supplemental treatments. However, little is known about levamisole's effects or its mechanism of action in horses. OBJECTIVE Levamisole in combination with another mitogen will stimulate a macrophage 1 (M1), dendritic cell 1 (DC1), T-helper 1 (CD4 Th1), and T-cytotoxic (CD8) immune response in equine peripheral blood mononuclear cells (PBMCs) in vitro as compared to mitogen alone. ANIMALS Ten neurologically normal adult horses serologically negative for Sarcocystis neurona. METHODS Prospective study. Optimal conditions for levamisole were determined based on cellular proliferation. Peripheral blood mononuclear cells were then cultured using optimal conditions of mitogen and levamisole to identify the immune phenotype, based on subset-specific activation markers, intracellular cytokine production, and cytokine concentrations in cell supernatants. Subset-specific proliferation was determined using a vital stain. RESULTS Concanavalin A (conA) with levamisole, but not levamisole alone, resulted in a significant decrease (P

Guided by a grounded theory methodology, the authors propose a theory of rehabilitation for incarcerated persons within a prison equine program (PEP). Interviews with ten incarcerated men yielded a grounded theory of rehabilitative change centered around the importance of relationships with program staff and horses, and the uniqueness of the barn-equine environment in promoting safe attachments and positive views of self. Special emphasis is placed on the development of secure reparative attachments between men and their horses grounded in acceptance and mutual empathy, as well as the development of redemptive identities whereby participants viewed themselves as having purpose and value as human beings. Implications for policy and practice for fostering effective rehabilitation in incarceration settings are discussed.

Neuroendocrine tumors are rarely diagnosed in horses. This report describes a case of a neuroendocrine tumor with strong similarities to descriptions of posterior mediastinal paragangliomas in humans and dogs. A 12-year-old Quarter Horse gelding was presented initially for management of hemothorax of unknown origin that responded to medical management. Nineteen months later, the horse was presented again with acute-onset hindlimb ataxia, at which time a thoracic mass adjacent to the vertebral bodies was detected on radiography. The gelding was euthanized after failing to respond to anti-inflammatory therapy, and on necropsy, the mass was identified as a paraganglioma invading the spinal canal. Despite its locally invasive behavior, the tumor showed no evidence of metastasis, and its apparent slow progression was in sharp contrast to more common thoracic neoplasms such as hemangiosarcoma. Combined with the reported success of surgical excision in human mediastinal paragangliomas, this suggests that early diagnosis of such tumors could provide the opportunity for successful treatment.

Introduction: Previous studies have shown that apparently healthy animals participating in Animal-Assisted Interventions (AAI) have the potential to asymptomatically carry and even transmit zoonotic pathogens to people, which is of particular concern for therapy animal teams visiting healthcare settings. This two-part study was designed to investigate the risk of zoonotic pathogen transmission within a university-based AAI program as a combination of the prevalence of these pathogens in the animal population as well as the handlers’ understanding of the risks of zoonoses in AAI and their adherence to infection control practices. Methods: In part one of the study, AAI program records were retrospectively reviewed and infectious disease screening test results were compiled from 22 dogs and 2 cats. Screening tests for dogs and cats included a zinc sulfate fecal float, fecal culture, and nasal and perianal skin swabs for methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudointermedius (MRSP). Additional tests for cats were blood cultures for Bartonella henselae and Toxoplasmosis IgG and IgM antibody titers. In part two, a survey was conducted of 40 registered therapy animal handlers to assess knowledge, attitudes, and perceptions regarding risk of infectious disease transmission in AAI settings, including risk to the animal, the handler, and those being visited. Results: In part one, there were 17 total positive results of the 118 infectious disease screenings performed, 14 of which were potentially zoonotic organisms. In part two of the study, a majority (70%) of respondents expressed they had no concerns regarding infectious disease transmission in AAI settings. Despite handler education and guidelines, adherence to infection control practices was lacking. Discussion: The results of this study support prior findings that animals participating in AAI can be asymptomatic carriers of zoonotic organisms. Compliance with infection control practices and hand hygiene are paramount to mitigate risk of zoonotic disease transmission, but was inconsistent among this group of handlers. Given the popularity of AAI programs in the U.S., similar studies should be performed on a larger scale to determine the level of adherence to currently recommended practices and potential need for improvement in infectious disease control education and/or policies. No funding was received to conduct the studies. Open access publication fees will be paid by the Small Animal Department at the Virginia-Maryland College of Veterinary Medicine.

Background Equine neuroborreliosis (NB), Lyme disease, is difficult to diagnose and has limited description in the literature. Objective Provide a detailed description of clinical signs, diagnostic, and pathologic findings of horses with NB. Animals Sixteen horses with histologically confirmed NB. Methods Retrospective review of medical records at the University of Pennsylvania and via an ACVIM listserv query with inclusion criteria requiring possible exposure to Borrelia burgdorferi and histologic findings consistent with previous reports of NB without evidence of other disease. Results Sixteen horses were identified, 12 of which had additional evidence of NB. Clinical signs were variable including muscle atrophy or weight loss (12), cranial nerve deficits (11), ataxia (10), changes in behavior (9), dysphagia (7), fasciculations (6), neck stiffness (6), episodic respiratory distress (5), uveitis (5), fever (2), joint effusion (2), and cardiac arrhythmias (1). Serologic analysis was positive for B. burgdorferi infection in 6/13 cases tested. CSF abnormalities were present in 8/13 cases tested, including xanthochromia (4/13), increased total protein (5/13; median: 91 mg/dL, range: 25–219 mg/dL), and a neutrophilic (6/13) or lymphocytic (2/13) pleocytosis (median: 25 nucleated cells/μL, range: 0–922 nucleated cells/μL). PCR on CSF for B. burgdorferi was negative in the 7 cases that were tested. Conclusion and Clinical Importance Diagnosis of equine NB is challenging due to variable clinical presentation and lack of sensitive and specific diagnostic tests. Negative serology and normal CSF analysis do not exclude the diagnosis of NB.

The innate immune response to lipopolysaccharide contributes substantially to the morbidity and mortality of gram-negative sepsis. Horses and humans share an exquisite sensitivity to lipopolysaccharide and thus the horse may provide valuable comparative insights into this aspect of the inflammatory response. MicroRNAs, small non-coding RNA molecules acting as post-transcriptional regulators of gene expression, have key roles in toll-like receptor signaling regulation but have not been studied in this context in horses. The central hypothesis of this study was that lipopolysaccharide induces differential microRNA expression in equine peripheral blood mononuclear cells in a manner comparable to humans. Illumina Next Generation Sequencing was used to characterize the basal microRNA transcriptome in isolated peripheral blood mononuclear cells from healthy adult horses, and to evaluate LPS-induced changes in microRNA expression in cells cultured for up to four hours. Selected expression changes were validated using quantitative reverse-transcriptase PCR. Only miR-155 was significantly upregulated by LPS, changing in parallel with supernatant tumor necrosis factor-α concentration. Eight additional microRNAs, including miR-146a and miR-146b, showed significant expression change with time in culture without a clear LPS effect. Target predictions indicated a number of potential immunity-associated targets for miR-155 in the horse, including SOCS1, TAB2 and elements of the PI3K signaling pathway, suggesting that it is likely to influence the acute inflammatory response to LPS. Gene alignment showed extensive conservation of the miR-155 precursor gene and associated promoter regions between horses and humans. The basal and LPS-stimulated microRNA expression pattern characterized here were similar to those described in human leukocytes. As well as providing a resource for further research into the roles of microRNAs in immune responses in horses, this will facilitate inter-species comparative study of the role of microRNAs in the inflammatory cascade during endotoxemia and sepsis. Published version

Summary Debate surrounding the nature of equine melanoma has resulted in an underestimation of its life-threatening potential. Contrary to popular dogma, the variable, often slow, rate of growth commonly associated with equine melanoma does not warrant benign classification. Equine melanoma is a malignant neoplasm with the capacity for local invasion and metastasis. A classification scheme was proposed in 1995, but this does not address the progressive nature of equine malignant melanoma (EMM). Additionally, frustration with conflicting therapeutic recommendations has led many practitioners to inappropriately advocate benign neglect. This article addresses the need for a clinically applicable, standardised classification system, provides a review of current therapies and recommendations for equine practitioners, and comments on the future directions of EMM research.

Objective—To measure the ascorbic acid (AA) concentration in bronchoalveolar lavage fluid (BALF) and cellular glutathione peroxidase (cGPx) activity in RBCs and WBCs from peripherally obtained blood and in cells from BALF to determine whether differences existed between the 2 major redox systems in recurrent airway obstruction (RAO)-affected and -nonaffected (control) horses and between systemic and local pulmonary responses in the glutathione redox system. Animals—16 adult horses in pairs: 8 healthy (control) and 8 RAO-affected horses. Procedures—Physical examination data and biological samples were collected from horses before (remission), during, and after (recovery) environmental challenge with dusty straw and hay. At each stage, BALF cell AA concentration and RBC, WBC, and BALF cell cGPx activity were measured. Results—Compared with control horses, RAO-affected horses had significantly higher cGPx activity in RBCs at all points and in WBCs during remission and challenge. The BALF cell cGPx activity was higher in RAO-affected horses during recovery than during remission The BALF cell AA concentration did not differ significantly in control horses at any point, but total and free AA concentrations were significantly lower in RAO-affected horses during the challenge period than during remission and recovery periods. Conclusions and Clinical Relevance—High cGPx activity suggested this redox system was upregulated during exposure to dusty straw and hay to combat oxidative stress, as AA was depleted in RAO-affected horses. The relative delay and lack of comparative increase in cGPx activity within the local environment (represented by BALF cells), compared with that in RBCs and WBCs, might contribute to disease in RAO-affected horses.

Objective—To evaluate the phospholipid composition and function of surfactant in horses with recurrent airway obstruction (RAO) at various clinical stages and compare these properties with findings in horses without RAO. Animals—7 horses with confirmed RAO and 7 without RAO (non-RAO horses). Procedures—Pairs of RAO-affected and non-RAO horses were evaluated before, during, and after exposure to hay. Evaluations included clinical scoring, lung function testing, airway endoscopy, and bronchoalveolar lavage fluid (BALF) absolute and differential cell counts. Cell-free BALF was separated into crude surfactant pellet and supernatant by ultracentrifugation, and phospholipid and protein concentrations were determined. Phospholipid composition of crude surfactant pellets and surface tension were evaluated with high-performance liquid chromatography and a pulsating bubble surfactometer, respectively. Findings were compared statistically via mixed-effects, repeated-measures ANOVA. Results—Total phospholipid concentration in BALF was lower in RAO-affected versus non-RAO horses at all sample collection times. In the RAO-affected group, total phospholipid concentration was lower during exposure to hay than before or after exposure. There were no significant differences in BALF protein concentration, percentages of phospholipid classes, or surface tension between or within groups of horses. Conclusions and Clinical Relevance—All clinical stages of RAO-affected horses were characterized by low surfactant concentration in BALF. Exacerbation of RAO led to an additional decrease in surfactant concentration. Causes for low surfactant concentration in RAO-affected horses remain to be determined. Low phospholipid concentration may render RAO-affected horses more susceptible than unaffected horses to surfactant alterations and contribute to clinical disease status and progression.

Porcine circovirus type 2 (PCV2) belongs to the viral family Circoviridae and to the genus Circovirus. Circoviruses are small, single-stranded nonenveloped DNA viruses that have an unsegmented circular genome. PCV2 is the primary causative agent of several syndromes collectively known as porcine circovirus-associated disease (PCVAD). Many of the syndromes associated with PCVAD are a result of coinfection with PCV2 virus and other agents such as Mycoplasma and porcine reproductive and respiratory syndrome virus. PCV2 infection is present in every major swine-producing country in the world, and the number of identified cases of PCVAD is rapidly increasing. In the United States, the disease has cost producers an average of 3-4 dollars per pig with peak losses ranging up to 20 dollars per pig. The importance of this disease has stimulated investigations aimed at identifying risk factors associated with infection and minimizing these risks through modified management practices and development of vaccination strategies. This paper provides an overview of current knowledge relating to PCV2 and PCVAD with an emphasis on information relevant to the swine veterinarian.

Objective—To evaluate the precision of intradermal testing (IDT) in horses. Animals—12 healthy adult horses. Procedure—IDT was performed on the neck of each horse by use of 2 positive control substances (histamine and phytohemagglutinin [PHA]) and a negative control substance. An equal volume (0.1 mL) for each injection was prepared to yield a total of 20 syringes ([4 concentrations of each positive control substance plus 1 negative control substance] times 2 positive control substances times 2 duplicative tests) for each side of the neck. Both sides of the neck were used for IDT; therefore, 40 syringes were prepared for each horse. Hair was clipped on both sides of the neck, and ID injections were performed. Diameter of the skin wheals was recorded 0.5, 4, and 24 hours after ID injection. Results—Intra- and interhorse skin reactions to ID injection of histamine and PHA resulted in wheals of uniform size at 0.5 and 4 hours, respectively. Significant intra- and interhorse variation was detected in wheals caused by PHA at 24 hours. Conclusions and Clinical Relevance—ID injection of histamine and PHA caused repeatable and precise results at 0.5 and 4 hours, respectively. Concentrations of 0.005 mg of histamine/mL and 0.1 mg of PHA/mL are recommended for use as positive control substances for IDT in horses. This information suggests that consistent wheal size is evident for ID injection of control substances, and variation in wheals in response to ID injection of test antigens results from a horse's immune response to specific antigens. (Am J Vet Res 2005;66:1341–1347)

Virulent strains of the bovine opportunistic pathogen Haemophilus somnus ( Histophilus somni ) cause multi-systemic diseases in cattle. One of the reported virulence factors that H. somnus may use to persist in the host is resistance to intracellular killing. We report here that H. somnus significantly ( P 2 − ) by bovine mammary and alveolar macrophages as well as by polymorphonuclear leukocytes. Inhibition of O 2 − was time- and dose-dependent and did not occur after incubation with Escherichia coli , H. influenzae , or Brucella abortus . Non-viable H. somnus , purified lipooligosaccharide, or cell-free supernatant from mid-log phase cultures did not inhibit O 2 − production, indicating that O 2 − inhibition required contact with live H. somnus . Furthermore, preincubation of phagocytic cells with cytochalasin B to prevent phagocytosis did not decrease the ability of H. somnus to inhibit O 2 − production. Some H. somnus isolates from the prepuce of healthy bulls were less capable or incapable of inhibiting macrophage O 2 − production compared to isolates tested from disease sites. Our results suggest that inhibition of O 2 − may be an important virulence factor exploited by pathogenic strains of H. somnus to resist killing by professional phagocytic cells.

Abstract The anorexic or dysphagic adult horse often requires nutritional support. Providing nutrients by the enteral route is the safest and most economic choice, but the dietary options available for use in horses are somewhat limited. The objective of this study was to compare the physiologic response of normal horses with a low-residue liquid or normal diet over a 10-day feeding period. Two groups of 6 normal adult horses were maintained on 1 of 2 diets for a 10-day period. Diets were formulated to meet the caloric needs of a horse maintained in a stall. The control group was fed 70% timothy hay and 30% textured concentrate for the test period, and the experimental group received the low-residue liquid diet, similar to liquid nutritional formulas designed for human use. Clinical parameters, body weight, packed cell volume, total plasma solids, blood glucose, and serum electrolytes were recorded daily for each horse during the dietary trial period. On days 1, 5, and 10 of the study, a complete blood count, serum biochemical profile, and urinalysis were performed. Horses' serum total bilirubin concentration and pattern of weight loss differed between groups. All other physical parameters, blood chemistry, complete blood count, and urinalysis results remained within the normal reference interval for the horses regardless of diet, although some statistical differences were observed. Horses returned to pasture and free-choice grass diet without complications at the end of the dietary trial period. These results demonstrate that few differences of biologic significance were observed between horses being fed low-residue diet and horses receiving a normal diet of hay and grain over a 10-day period. (Equine Vet J 2003;23:310–317)

Objective—To determine whether antibodies against Sarcocystis neurona could be detected in CSF from clinically normal neonatal (2 to 7 days old) and young (2 to 3 months old) foals. Design—Prospective study. Animals—15 clinically normal neonatal Thoroughbred foals. Procedure—Serum and CSF samples were obtained from foals at 2 to 7 days of age and tested for antibodies against S neurona by means of western blotting. Serum samples from the mares were also tested for antibodies against S neurona. Additional CSF and blood samples were obtained from 5 foals between 13 and 41 days after birth and between 62 and 90 days after birth. Results—Antibodies against S neurona were detected in serum from 13 mares and their foals; antibodies against S neurona were detected in CSF from 12 of these 13 foals. Degree of immunoreactivity in serum and CSF decreased over time, and antibodies against S neurona were no longer detected in CSF from 2 foals 83 and 84 days after birth. However, antibodies could still be detected in CSF from the other 3 foals between 62 and 90 days after birth. Conclusions and Clinical Relevance—Results indicate that antibodies against S neurona can be detected in CSF from clinically normal neonatal (2 to 7 days old) foals born to seropositive mares. This suggests that western blotting of CSF cannot be reliably used to diagnose equine protozoal myeloencephalitis in foals < 3 months of age born to seropositive mares. (J Am Vet Med Assoc 2002;220:208–211)

Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM), affecting 0.5–1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.

Neonatal alloimmune thrombocytopenia is recognized as a spontaneous disease of human infants, piglets, and possibly mules, but it has not been previously reported in horses. A 1-day-old Quarter Horse foal presented to Michigan State University Large Animal Clinic with severe thrombocytopenia of unknown origin. Immunoglobulins that bound to the foal's platelets were identified in the mare's plasma, serum, and milk by indirect assays. The immunoglobulins were further shown to recognize platelets from the foal's full brother, born 1 year earlier. These findings, coupled with the clinical course of the foal during its period of hospitalization, strongly suggest that neonatal alloimmune thrombocytopenia can spontaneously occur in neonatal horses. This diagnosis should be considered for foals with severe thrombocytopenia when other causes can be excluded, and platelet antibody assays should be used to support this diagnosis.

Objective—To quantify peripheral blood neutrophil apoptosis in equine patients with acute abdominal disease (ie, colic) caused by strangulating or nonstrangulating intestinal lesions and compare these values with values for horses undergoing elective arthroscopic surgery. Animals—20 client-owned adult horses. Procedures—Peripheral blood was collected from horses immediately prior to and 24 hours after surgery for treatment of colic (n = 10) or elective arthroscopic surgery (10), and neutrophils were counted. Following isolation by means of a bilayer colloidal silica particle gradient and culture for 24 hours, the proportion of neutrophils in apoptosis was detected by flow cytometric evaluation of cells stained with annexin V and 7-aminoactinomycin D. Values were compared between the colic and arthroscopy groups; among horses with colic, values were further compared between horses with and without strangulating intestinal lesions. Results—Percentage recovery of neutrophils was significantly smaller in preoperative samples (median, 32.5%) and in all samples combined (35.5%) for the colic group, compared with the arthroscopy group (median, 66.5% and 58.0%, respectively). No significant differences in the percentages of apoptotic neutrophils were detected between these groups. Among horses with colic, those with strangulating intestinal lesions had a significantly lower proportion of circulating apoptotic neutrophils in postoperative samples (median, 18.0%) than did those with nonstrangulating lesions (66.3%). Conclusions and Clinical Relevance—The smaller proportion of apoptotic neutrophils in horses with intestinal strangulation suggested that the inflammatory response could be greater or prolonged, compared with that of horses with nonstrangulating intestinal lesions. Further investigations are needed to better understand the relationship between neutrophil apoptosis and inflammation during intestinal injury.

Summary Transtracheal wash and bronchoalveolar lavage are diagnostic techniques that have been adopted from human medicine for monitoring inflammatory changes in the airway of the horse. Transendoscopic biopsy has also proven to be a valuable tool for obtaining samples of the airway mucosa in human patients. A transendoscopic technique was developed in this study for obtaining a respiratory mucosal biopsy from standing, sedated horses. Six normal adult horses were sampled at eight-week intervals for a total of three sample periods. Horses were monitored for adverse effects of the technique and none were noted. Sample sites were completely healed after eight weeks with no gross or histologic abnormalities. Biopsy samples were 3 to 4 millimeters in diameter, and 17 of 18 samples provided interpretable histological sections. Methods for handling, staining and evaluating tissue were also developed. The results of this study demonstrated that airway mucosal biopsy is a safe, repeatable technique that can be performed in the sedated, standing horse.

Recurrent airway obstruction (RAO, also known as equine heaves) is an inflammatory condition similar to human asthma caused by exposure of susceptible horses to poorly ventilated stable environments. The disease is characterized by neutrophilic airway inflammation, mucus hypersecretion and reversible bronchoconstriction. This inflammatory process is mediated by several factors, including antibodies, cytokines, resident cells of the airway and inflammatory cellular components that arrive in the respiratory tract. An increasing body of evidence has lent support to the concept that a dysregulation of T cell apoptosis may play a central role in the development of airway inflammation and the associated asthma. Therefore, the aim of this study was to investigate early and late apoptosis of CD4 and CD8 T cell subpopulations obtained from the airways of acute RAO-positive animals after exposure to hay/straw. The percentages of CD4 and CD8 T cells and their associated frequencies of apoptosis were quantified using flow cytometry. Hay/straw exposure induced clinical airway obstruction, airway neutrophilia and increased airway mucus production in RAO-positive horses. In addition, allergen exposure increased the percentage of CD4 T cells in RAO-positive horses as well as the frequency of early and late apoptosis in both CD4 and CD8 lymphocyte subpopulations. These results suggest that the higher frequency of lymphocyte apoptosis may play a role in disease progression of horses afflicted with RAO and may partially explain the characteristic remission of this pathological condition once the allergen source is removed. However, further studies are needed to clarify the role of T cell apoptosis in RAO-affected horses.

Background: Surfactant alterations are described in horses after exercise, anesthesia, and prolonged transport, in horses with recurrent airway obstruction, and in neonatal foals. The effect of horse age or bronchoalveolar lavage fluid (BALF) sample characteristics on surfactant is unknown. Objectives: To evaluate surfactant phospholipid composition and function in healthy horses, and to investigate the influence of age and BALF sample characteristics on surfactant. Animals: Seventeen healthy horses 6–25 years of age maintained on pasture year-round. Methods: BALF was collected by standard procedures and was assessed for recovery volume, nucleated cell count (NCC), and cytology. Cell-free BALF was separated into crude surfactant pellet (CSP) and surfactant supernatant (Supe) by ultracentrifugation. Phospholipid and protein content were determined from both fractions. CSP phospholipid composition was analyzed by high-performance liquid chromatography with an evaporative light scatter detector. Surface tension of CSP was evaluated with a pulsating bubble surfactometer. Regression analysis was used to evaluate associations between age, BALF sample characteristics, and surfactant variables. Results: Results and conclusions were derived from 15 horses. Increasing age was associated with decreased phospholipid content in CSP but not Supe. Age did not affect protein content of CSP or Supe, or surfactant phospholipid composition or function. Age-related surfactant changes were unaffected by BALF recovery percentage, NCC, and cytological profile. Conclusions and Clinical Importance: Older horses have decreased surfactant phospholipid content, which might be because of age-related pulmonary changes. Surfactant composition is unaffected by BALF sample characteristics at a BALF recovery percentage of at least 50%.

Lung surfactant is produced by type II alveolar cells as a mixture of phospholipids, surfactant proteins, and neutral lipids. Surfactant lowers alveolar surface tension and is crucial for the prevention of alveolar collapse. In addition, surfactant contributes to smaller airway patency and improves mucociliary clearance. Surfactant-specific proteins are part of the innate immune defense mechanisms of the lung. Lung surfactant alterations have been described in a number of respiratory diseases. Surfactant deficiency (quantitative deficit of surfactant) in premature animals causes neonatal respiratory distress syndrome. Surfactant dysfunction (qualitative changes in surfactant) has been implicated in the pathophysiology of acute respiratory distress syndrome and asthma. Analysis of surfactant from amniotic fluid allows assessment of fetal lung maturity (FLM) in the human fetus and exogenous surfactant replacement therapy is part of the standard care in premature human infants. In contrast to human medicine, use and success of FLM testing or surfactant replacement therapy remain limited in veterinary medicine. Lung surfactant has been studied in large animal models of human disease. However, only a few reports exist on lung surfactant alterations in naturally occurring respiratory disease in large animals. This article gives a general review on the role of lung surfactant in respiratory disease followed by an overview of our current knowledge on surfactant in large animal veterinary medicine.

The purpose of this consensus statement is to provide a review of current knowledge and opinions concerning inflammatory airway disease (IAD) and to help practitioners differentiate IAD from heaves (or recurrent airway obstruction; RAO) and other inflammatory respiratory diseases of horses.

Objective—To evaluate differences in response to ID injection of histamine, phytohemagglutinin (PHA), and Aspergillus organisms between clinically normal horses and horses with recurrent airway obstruction (RAO). Animals—5 healthy adult horses and 5 adult horses with RAO. Procedure—Intradermal testing (IDT) was performed on the neck with 2 positive control substances (histamine and PHA) and a mixture comprising 5 Aspergillus species. Four concentrations of each test substance plus a negative control substance were used. Equal volumes (0.1 mL) of each test substance were prepared to yield 15 syringes ([4 concentrations of each test substance plus 1 negative control substance] times 3 test substances) for each side of each horse (ie, 30 syringes/horse). Intradermal injections were administered; diameter of wheals was recorded 0.5, 4, and 24 hours after injection. Results—Hypersensitive responses to ID injection of histamine were detected 0.5 hours after injection, and a delay in wheal formation after ID injection of Aspergillus mixture 24 hours after injection was detected in RAO-affected horses but was not observed in clinically normal horses. No differences were detected between the 2 groups after ID injection of PHA. Conclusions and Clinical Relevance—RAO-affected horses are hypersensitive to histamine, suggesting that RAO is associated with a heightened vascular response to histamine. Higher concentrations of Aspergillus mixture may be needed to detect horses that are sensitive to this group of antigens. Wheal reactions to Aspergillus may be a delayed response, suggesting that IDT results should be evaluated 0.5, 4, and 24 hours after ID injection. (Am J Vet Res2005;66:1348–1355)

In recent years, equine neonatal medicine has made significant advances. The importance of nutritional support for the sick neonatal foal has been recognized, and methods of providing that sup-port have been developed. Today, the clinician has many options when designing a nutritional plan for the neonatal foal. When the foal's gut permits, enteral diets are an inexpensive source of nutrients. Under conditions where the gut requires rest, methods for delivering nutrients by the parenteral route have also been developed. In this article, the nutrition of the normal and sick foal is described. Guidelines for designing a nutritional plan are also reviewed.

Objective—To determine whether immune function can be accurately assessed in blood samples obtained from horses and refrigerated overnight and whether a nonradioactive lymphocyte proliferation assay can be used to evaluate samples obtained from horses. Sample Population—224 blood samples from 28 clinically normal adult horses. Procedure—Heparinized blood samples were collected. Each sample was divided into 2 equal aliquots. One aliquot was refrigerated overnight to simulate overnight shipping of blood samples, and the other aliquot was evaluated on the day of blood collection. Lymphocytes were isolated and enumerated by use of a modified single-gradient procedure. Cell viability and function were assessed by use of cytologic examination, flow cytometry, and mitogen-induced proliferation assays. Lymphocyte proliferation in response to T- and B-cell mitogens was measured by use of [3H]-thymidine incorporation and a nonradioactive lymphocyte proliferation assay. Results—Lymphocytes refrigerated for up to 24 hours continued to be acceptable for use in immunologic analysis on the basis that they maintained viability and did not have significant alterations in lymphocyte subsets, except for CD8, when compared with freshly isolated lymphocytes. Furthermore, results for mitogeninduced lymphocyte proliferation assays were also comparable between fresh and refrigerated aliquots. Conclusions and Clinical Relevance—The nonradioactive lymphocyte proliferation assay is a reliable alternative to [3H]-thymidine assay for assessing proliferation of equine lymphocytes. Collectively, our results imply that blood samples refrigerated and shipped overnight to a laboratory can be used to perform cellular-immune assays; results of those assays would enhance a clinician's diagnostic abilities to monitor the efficacy of treatment. (Am J Vet Res 2003;64:1003–1009)

Objective—To identify apoptosis in equine intestines and determine whether apoptosis is associated with gastrointestinal tract disease or a specific tissue layer of intestine.Animals—38 horses that underwent surgery or were euthanatized for small or large intestine obstruction, strangulation, or distension and 9 control horses euthanatized for reasons other than gastrointestinal tract disease or systemic disease.Procedure—Specimens were collected at surgery from intestine involved in the primary lesion and distant to the primary lesion site or at necropsy from several sites including the primary lesion site. Histologic tissue sections were stained with H&E, and apoptosis was detected by use of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling technique. The number of apoptotic cells per hpf was counted in the mucosa, circular muscle, longitudinal muscle, and serosa.Results—Apoptotic nuclei were seen in all layers of intestine. An increased number of apoptotic cells was found in the circular muscle of the intestine from horses with simple obstruction, compared with strangulating obstruction or healthy intestine. Intestine distant from a primary strangulating lesion had higher numbers of apoptotic cells than did intestine distant from a simple obstructive lesion or intestine taken at the site of a strangulating or simple obstructive lesion.Conclusions and Clinical Relevance—Intestine from horses with obstructing or strangulating lesions in the small intestine and large colon had high numbers of apoptotic cells possibly because of ischemic cell injury and subsequent inflammation. Whether substantial apoptosis affects intestinal function is not yet known. (Am J Vet Res2003;64:982–988)

Horses that are exposed to Sarcocystis neurona, a causative agent of equine protozoal myeloencephalitis, produce antibodies that are detectable in serum by western blot (WB). A positive test is indicative of exposure to the organism. Positive tests in young horses can be complicated by the presence of maternal antibodies. Passive transfer of maternal antibodies to S. neurona from seropositive mares to their foals was evaluated. Foals were sampled at birth (presuckle), at 24h of age (postsuckle), and at monthly intervals. All foals sampled before suckling were seronegative. Thirty-three foals from 33 seropositive mares became seropositive with colostrum ingestion at 24h of age, confirming that passive transfer of S. neurona maternal antibodies occurs. Thirty-one of the 33 foals became seronegative by 9 months of age, with a mean seronegative conversion time of 4.2 months. These results indicate that evaluation of exposure to S. neurona by WB analysis of serum may be misleading in young horses.

Horses affected with Recurrent Airway Obstruction (RAO) exhibit inflammation and oxidative damage to lung cells. The main objective of this study was to develop a novel antioxidant delivery system (ADS) capable of targeting activated macrophages within the lower airways in order to reduce cellular oxidative damage. Specifically, this ADS consisted of folic acid and glutathione (GSH) linked to bisamine poly(ethylene oxide) (PEO). Folic acid conjugates exploit the over-expression of folate receptors on activated macrophages, allowing for the internalization of the ADS through receptor-mediated endocytosis. Initial results from in vitro studies demonstrated active targeting and uptake through folate receptor mediated endocytosis. GSH concentration in glutathione deficient RAW 264.7 cells (an immortalized murine macrophage cell line) was increased by more than 300% after incubation with the ADS for four hours effectively restoring 50% of the normal GSH concentration. F2-Isoprostanes, biomarkers for lipid oxidation, were reduced in RAW 264.7 cells by more than 67% after incubation with the ADS and exposure to zymosan, a free radical initiator. Evaluation of ADS as a treatment for equine RAO is underway and the results will be included in this presentation.

Pulmonary immune responses are suited to determine mechanisms of lymphocyte elimination, as lung inflammation must be regulated tightly to preserve gas exchange. The self-terminating response of primed C57BL/6 mice to intratracheal challenge with the T cell-dependent Ag sheep erythrocytes (SRBC) was used to test the importance of lung lymphocyte apoptosis in pulmonary immunoregulation. Apoptosis of alveolar and interstitial lymphocytes was demonstrated morphologically, by three independent methods to detect DNA fragmentation, and by surface expression of phosphatidylserine. Apoptotic lymphocytes were exclusively CD4-, CD8-, B220-, but many were CD3+ and Thy 1+. Inhibiting apoptosis by in vivo cyclosporine treatment prolonged lung lymphocyte accumulation following SRBC challenge. Experiments using mice homozygous for the lpr or gld mutations showed that pulmonary lymphocyte apoptosis depended on expression of Fas (CD95) and its ligand (Fas-L). Pulmonary inflammation increased on repeated intratracheal SRBC challenge of lpr/lpr mice, in contrast to the waning response in normal mice. These results confirm that in situ lymphocyte apoptosis contributes to termination of immune responses in nonlymphoid organs, probably because of activation-induced cell death, and may be important in inducing tolerance to repeated antigen exposure.

Leukemia is a neoplastic disease of one or more of the cell types of the hemopoietic system and is rarely diagnosed in the horse. This report describes a case of subleukemic acute myelomonocytic leukemia in an 11 -year-old gelding. Preliminary cytological diagnosis was supported by two types of laboratory investigations. Cytochemical characterization of blood and bone marrow neoplastic cells was consistent with a myelomonocytic origin. Neoplastic blast cells in peripheral blood were labeled by monoclonal antibodies specific for cell surface molecules of horse granulocytes, but they were not labeled by antibodies to T- or B-lymphocytes or macrophages. Treatment was attempted but was unsuccessful. At necropsy, intravascular leukostasis was present in all tissues examined. Fungal hyphae were also found in lung interstitium and colonic submucosa, suggesting the presence of a systemic mycosis. Nucleated cells were isolated from peripheral blood and cultured in vitro; they survived for up to 2 weeks and had evidence of cell division that was not sustained. Frozenthawed cells stored in liquid nitrogen were also successfully cultured in vitro, but no permanent cell lines could be established.

A vaccine against Sarcocystis neurona, which induces equine protozoal myeloencephalitis (EPM), has received conditional licensure in the United States. A major concern is whether the immunoglobulin G (IgG) response elicited by the vaccine will compromise the use of Western blotting (WB) as a diagnostic tool in vaccinated horses with neurologic disease. Our goals were to determine if vaccination (1) causes seroconversion: (2) causes at least a transient increase in S neurona-specific IgG in the cerebrospinal fluid (CSF); and (3) induces an IgG response that can be differentiated from that induced by natural exposure. Horses included in the study (n = 29) were older than 6 months with no evidence of neurologic disease. The presence or absence of anti-S neurona antibodies in the serum of each horse was determined by WB analysis. Seropositive horses had CSF collected and submitted for cytology, CSF index, and WB analysis. The vaccine was administered to all the horses and boostered 3-4 weeks later. On day 14 after the 2nd administration, serum and CSF were collected and analyzed. Eighty-nine percent (8 of 9) of the initial seronegative horses seroconverted after vaccination, of which 57% (4 of 7) had anti-S neurona IgG in their CSE Eighty percent (16 of 20) of the seropositive horses had an increase in serum S neurona IgG after vaccination. Of the 6 of 20 horses that were initially seropositive/CSF negative, 2 were borderline positive for anti-S neurona IgG in the CSF, 2 tested positive, and 2 were excluded because the CSF sample had been contaminated by blood. There were no WB banding patterns that distinguished samples from horses that seroconverted due to vaccination versus natural exposure. Caution must be used in interpreting WB analysis from neurologic horses that have been recently vaccinated for EPM.

a b s t r a c t The effect of animal-assisted activities (AAA) on the animal participants has been mini- mally investigated, and the welfare of these animals has been questioned. To enhance our understanding of these animals' welfare, we measured cortisol collected from serial saliva samples of 15 healthy adult dogs registered with an AAA organization. We collected saliva every 30 min before, during, and after a standardized 60-min session across three sett- ings: an AAA session (activity) for college students in the communal area of a residence hall, a novel session located in a novel room without interaction with a stranger, and a home session inside each handler's own home. Each session was videotaped, and specific behaviors during 5-min petting interactions were coded. Salivary cortisol levels were sig- nificantly higher in the novel setting (0.397 g/dL) compared to activity (0.257 g/dL) and home (0.213 g/dL) settings at time 30 min (P = 0.01 and P = 0.03, respectively). Dogs exhib- ited significantly more standing (59% vs 0%, P = 0.008) and ambulating (5.6% vs 0%, P = 0.001) behavior in the activity setting compared to the home at time 30 min, as well. Salivary corti- sol level was negatively correlated with panting (P = 0.02) and standing (P = 0.02) at specific time points in the novel and activity settings, respectively. During the 60-min AAA session, salivary cortisol concentration and stress-associated behavior were not statistically differ- ent compared to when dogs spent the same amount of time in the home setting, suggesting that they were not distressed when participating in the AAA sessions. The predictability of the environment may be an important consideration when evaluating the effect of AAA on