Multiple hypersensitivities including recurrent airway obstruction, insect bite hypersensitivity, and urticaria in 2 warmblood horse populations.

BACKGROUND: Multiple hypersensitivities (MHS) have been described in humans, cats, and dogs, but not horses. HYPOTHESES: Horses suffering from recurrent airway obstruction (RAO), insect bite hypersensitivity (IBH), or urticaria (URT) will have an increased risk of also being affected by another one of these hypersensitivities. This predisposition for MHS also will be associated with decreased shedding of strongylid eggs in feces and with a single nucleotide polymorphism (SNP BIEC2-224511), previously shown to be associated with RAO. ANIMALS: The first population (P1) included 119 randomly sampled horses representative of the Swiss sporthorse population; the replication population (P2) included 210 RAO-affected Warmblood horses and 264 RAO-unaffected controls. All horses were Warmbloods, 14 years or older. METHODS: Associations between disease phenotypes (RAO, IBH, URT, MHS) fecal egg counts, the SNP BIEC2-224511 as well as management and environmental factors were investigated. RESULTS: In P1, RAO-affected horses had a 13.1 times higher odds ratio (OR) of also suffering from IBH (P = .004). In P2, the respective OR was 7.4 (P = .002) and IBH-affected horses also showed a 7.1 times increased OR of concomitantly suffering from URT (P < .001). IBH, URT, and MHS phenotypes were significantly associated with the absence of nematode eggs in the feces. CONCLUSIONS AND CLINICAL IMPORTANCE: This is the first report of MHS in horses. Specifically, an increased risk for IBH should be expected in RAO-affected horses.

Venereal shedding of equid herpesvirus-1 (EHV-1) in naturally infected stallions.

BACKGROUND: Equid herpesvirus 1 (EHV-1) is a highly prevalent pathogen in horse populations worldwide. Oronasal infection represents the classic route of disease transmission. Venereal shedding of EHV-1 is not regarded relevant in terms of virus spreading, which is in contrast to the close relatives of EHV-1, bovine and suid alphaherpesvirus, for which artificial insemination is a well-documented and accepted means of virus spread. OBJECTIVES: Documentation of venereal EHV-1 shedding in 3 naturally infected stallions. ANIMALS: Three stallions were infected during an acute outbreak by an EHV-1 strain with the G(2254) /D(752) Pol genotype. METHODS: In this observational study, 12 semen samples from these 3 stallions were tested for EHV-1 to determine venereal shedding. EHV-1 was diagnosed by conventional PCR and paired serum neutralization tests in 42 horses. Semen samples were separated into sperm and seminal plasma fractions and tested for EHV-1 by conventional and quantitative PCR as well as virus isolation by cell culture. RESULTS: Acute EHV-1 infection was diagnosed on the premise. Five semen samples collected from 2 of the 3 stallions tested positive for EHV-1 by (q)PCR. On days 18 and 20 after onset of fever, the last positive samples were retrieved. All samples were positive in seminal plasma, only three in sperm fraction. Virus isolation attempts were unsuccessful. CONCLUSIONS AND CLINICAL IMPORTANCE: The data presented here document shedding of EHV-1 in semen of naturally infected stallions for close to 3 weeks, which seems not to be directly associated with spermatozoa.

[Select changes in the upper airways of the horse - an overview]. / Ausgewählte Veränderungen in den oberen Atemwegen des Pferdes - ein Uberblick.

Horses are obligate nasal breathers and depend on patency of their nasal passages. Several dynamic obstructive diseases in the pharyngeal and laryngeal area can be differentiated by high speed treadmill endoscopy and may be responsible for impaired exercise tolerance in the equine athlete. The anatomical specialty of guttural pouches predisposes the horse to species-specific diseases.

Second-generation microstimulator.

The first-generation injectable microstimulator was glass encased with an external tantalum capacitor electrode. This second-generation device uses a hermetically sealed ceramic case with platinum electrodes. Zener diodes protect the electronics from defibrillation shocks and from electrostatic discharge. The capacitor is sealed inside the case so that it cannot be inadvertently damaged by surgical instruments. This microstimulator, referred to as BION, is the main component of a 255-channel wireless stimulating system. BION devices have been implanted in rats for periods of up to 5 months. Results show benign tissue reactions resulting in identical encapsulation around BION and controls. Stimulation threshold levels did not change significantly over time and ranged between 0.81 to 1.35 mA for all the animals at a 60 micros pulse width. All of the tests performed to date indicate that the BION is safe and effective for long-term human implant. We have elected to develop BION applications by seeking collaboration with the research community through our BION Technology Partnership.

[Percutaneous occlusion of arterial vessels using permanent embolization for the treatment of an air sac hemorrhage in horses. A case report]. / Perkutaner Verschluss arterieller Gefässe durch permanente Embolisate zur Therapie einer Luftsackblutung beim Pferd. Ein Fallbericht.

This article reports a case of guttural pouch bleeding which was managed successfully by using intravascular embolisation systems to occlude the damaged vessels. Percutaneous catheterisation of the common carotid artery allowed angiographic visualisation of the main head arteries: A. carotis externa, A. carotis interna and A. occipitalis, which showed no abnormalities angiographically. Originating from the A. occipitalis, one artery sent smaller, extensively branching and tortuous vessels to the guttural pouch area. This branching was interpreted as a sign of inflammatory hypervascularization. The artery was occluded by positioning of a detachable balloon distally of the origin of the two small vessels. Insertion of two coils in the A. occipitalis proximally of the origin of this artery completed the embolisation. Occlusion of the distal part was necessary to avoid blood supply to the lesion from the contralateral arterial system via the circle of Willis. The technique used allowed occlusion of a selected head artery without direct surgical intervention in this area. There is no need to remove the implants. After catheterisation, no more episodes of epistaxis occurred. The causing diphtheroid inflammation in the guttural pouch was treated by local infusions of iodine-solutions and resolved completely within 24 days.

[Equine Cushing syndrome (ECS). Case report, review of its diagnosis and therapy and substantial differences from Cushing syndrome in dogs]. / Zum equinen Cushing-Syndrom (ECS). Fallbericht, Literaturauswertung zu Diagnostik und Therapie sowie wesentliche Unterschiede zum Cushing-Syndrome des Hundes.

Equine and canine Cushing's syndrome, both of which are the result of elevated cortisol levels, show some different pathogenetical and clinical features and require different therapeutical approaches. In older horses the equine Cushing's syndrome (ECS) is not uncommon. Nearly all cases result from excessive hormone production in cells of the pars intermedia of the pituitary. Besides elevated levels of adrenocorticotrope hormone (ACTH), high peripheral levels of pro-opiomelanocortin, beta-endorphines and alpha-melanocyte-stimulating hormone can be measured. In middle-aged and geriatric dogs, Cushing's syndrome is the most frequently diagnosed endocrinologic abnormality. 80-85% of cases are pituitary-dependent and 15-20% are caused by cortisol producing tumors of the adrenals. 90% of pituitary lesions can be identified as adenomas, which are localised in most cases in the pars distalis of the gland, but may occur rarely in the pars intermedia, too. Clinical symptoms in both species are characterised by wasting despite good appetite or polyphagia, reduction of muscle mass with altered fat deposition and lethargy. Whereas polydipsia/polyuria is a very common feature in dogs with Cushing's syndrome, in horses it is almost invariably a sign of concurrent secondary diabetes mellitus. A typical symptom in ECS is a continuously growing haircoat (hirsutism), whereas in canine Cushing's syndrome generalised alopecia may bring the owner to consult a veterinarian. The symptoms and diagnostic procedures in a 33-year-old mare are described. Useful diagnostic tests are reviewed with special attention to species differences in reacting to them. The therapeutic approach with dopamine-agonists such as bromocriptine and pergolide as well as cyproheptadine to ECS is reviewed.

[Demonstration of activity of two potentiated sulfonamides in feces of horses after oral or intravenous administration]. / Aktivitätsnachweis zweier potenzierter Sulfonamide in Kotproben von Pferden nach oraler oder intravenöser Verabreichung.

Both, the oral and intravenous application of two trimethoprim-potentiated sulfonamides induced measurable antibacterial activities in the feces of horses. With regard to the risk of antibiotic-induced alterations of the gastrointestinal flora, the route of application of potentiated sulfonamides seems to be of minor importance. The antibiotics used were Sulfadimethoxine/Trimethoprim (Trafigal 30% ad us. vet.) for oral and Sulfadoxine/Trimethoprim (Borgal 24% ad us. vet., both Hoechst AG, Frankfurt) for intravenous application. As recommended, both drugs were given in a dose of 20 mg per kg bodyweight. The detection method is based on a procedure layed down in German laws for sulfonamide residues in meat-samples and has undergone some modifications for the examination of feces.

Equine adenocarcinomas of the large intestine with osseous metaplasia.

Large intestinal adenocarcinoma with osseous metaplasia was diagnosed in two horses, a 15-year-old standard bred gelding and a 9-year-old Haflinger mare. Clinically, both animals had displayed weight loss and anaemia. A presumptive diagnosis of abdominal neoplasia was made and the horses were humanely killed. At necropsy, the gelding and the mare were found to have ulcerated tumours growing into the lumen of the caecum and colon, respectively. In the mare, the mass extended through the mesocolon and was evident in the left dorsal and ventral colon. Histopathologically, the tumours consisted of well-differentiated cords of single-layered columnar to cuboidal epithelial cells. Mitotic figures were very uncommon. In both lesions, well-formed bony spicules and osteoid were present in the fibrovascular stroma. The tumours were well-demarcated from surrounding mucosal tissue but had invaded the intestinal wall. Metastases were not observed.

[Susceptibility of bacterial isolates from the equine respiratory tract to trimethoprim, sulfadoxine, sulfadimethoxine and combinations of these compounds]. / Empfindlichkeit bakterieller Krankheitserreger aus dem Respirationstrakt von Pferden gegenüber Trimethoprim, Sulfadoxin, Sulfadimethoxin und Kombinationen dieser Wirkstoffe.

Using a broth microdilution technique, the in vitro susceptibility of bacterial isolates from the equine respiratory tract to trimethoprim, sulfadoxine, sulfadimethoxine, and combinations of these compounds was determined. The bacterial strains (n = 88) isolated recently from horses with respiratory symptoms belonged to the following species: Streptococcus equi subsp. zooepidemicus (n = 34), Streptococcus equi subsp. equi (n = 22), Staphylococcus aureus (n = 9), Klebsiella pneumoniae (n = 7), Rhodococcus equi (n = 4), Pseudomonas spp. (n = 3) and Escherichia coli (n = 3). In addition, two isolates of Enterobacter spp. and one isolate of Streptococcus equisimilis, Staphylococcus intermedius, Proteus mirabilis and Serratia marcescens were examined. For determination of susceptibility of an organism the following minimal inhibitory concentrations (MIC) were fixed as limiting values: Trimethoprim < or = 0.5 microgram/ml, sulfadoxine < or = 32 micrograms/ml, sulfadimethoxine < or = 32 micrograms/ml, trimethoprim/sulfadoxine < or = 0.5/32 micrograms/ml, trimethoprim/sulfadimethoxine < or = 0.5/32 micrograms/ml. As expected, Rhodococcus-equi-isolates were resistant to the antimicrobials tested. However, most of the clinically more common isolates showed a high degree of susceptibility to the combinations. The fractional inhibitory concentration (FIC) indices indicated synergism of the combination-partners in a wide range. According to these in vitro results, application of trimethoprim/sulfonamide combinations for the initial therapy of equine respiratory tract infections can be recommended.

[Demonstration of two trimethoprim/sulfonamide combinations in bronchoalveolar lavage fluid of horses and determination of blood levels]. / Nachweis zweier Trimethoprim/Sulfonamid-Kombinationen im Bronchialsekret von Pferden und Bestimmung der Blutspiegel.

Five healthy horses were given a sulfadoxine/trimethoprim combination (Borgal, Hoechst AG) i.v. on day 1. The next ten days the horses got once a day a sulfadimethoxine/trimethoprim combination orally (Trafigal, Hoechst AG). The doses were given as recommended. One horse received no medicaments for control. On each horse six bronchoalveolar lavages were performed. Blood samples were taken to calculate blood levels and elimination half lives. To determine the amount of substances in lavage fluid and plasma the high performance liquid chromatography (HPLC) was used. Regularly low quantities of sulfonamides and trimethoprim were detected in lavage-samples. The mean plasma concentration (n = 4) of sulfadoxine and trimethoprim 30 min after i.v. administration was 71.6 and 1.13 micrograms/g respectively. 24 h after injection the sulfadoxine blood level was 3.0 micrograms/g, while trimethoprim was no longer detectable. The average elimination half lives of sulfadoxine and trimethoprim were 7.94 h and 1.35 h respectively. 8 h after oral application (n = 5) the highest mean sulfadimethoxine blood levels of 53.8 micrograms/g were measured. The elimination half life of sulfadimethoxine was 9.77 h. Two hours after feeding the drug the first blood samples were taken. They already contained the highest mean trimethoprim concentration of 0.32 microgram/g plasma.

Cluster formation in a symmetrical network: a dynamical system for the description of the suppression among non-immune T lymphocytes and its application to the effects of immunization.

A mathematical model has been developed for the description of the suppressive regulation between polyclonally activated normal and immune T cells. The model assumes reversible cell-cell interactions to interpret results from limiting dilution experiments performed to determine the frequencies of precursor cells for antigen-specific T effector lymphocytes and to analyse mechanisms regulating the maturation of precursor into effector T cells. In particular, the model deals with the changes induced in the T lymphocytes population following immunization with antigens. In these limiting dilution experiments, T cells are placed in cultures at varying cell numbers with all other essential culture constituents kept in excess. After polyclonal activation of the T cells in culture they are supplied with growth and maturation factors so that they form daughter clones of functionally active T effector cells. The typical result observed was that effector T cells develop in cultures at low cell input but that this development is totally suppressed at high cell numbers. This result suggested that, at high cell numbers, the effector T cells are exposed to a sufficient number of other T cells of appropriate specificity to permit suppressive interactions. Whereas this is the case for non-immune T cells, T cells after immunization develop into effector cells both at high as well as at low cell concentrations, though with efficiencies less than proportional to their number of precursors. Our mathematical model is made up of a set of first order autonomous ordinary differential equations in many variables permitting the calculations of numbers of free cells and of cells engaged in cellular clusters of varying sizes. Free cells can develop into effector cells whereas cells engaged in clusters cannot. We calculate the consequences of several reasonable hypotheses concerning the effects of immunization. We consider the possibility that immunization modifies the growth behavior of the antigen-specific cells to permit an increased or accelerated clonal expansion in culture. Alternatively, we consider the possibility that immunization changes the interaction strength between cells specific for the immunizing antigen and other cells. Thirdly, we have connected both behaviors by calculating the case of an inverse relationship between growth rates and intensities of interaction between cells. Our model has been inspired by the symmetrical network model and can be interpreted in this framework. It proposes that immune regulation is a consequence of idiotype-anti-idiotype interactions.(ABSTRACT TRUNCATED AT 400 WORDS)

Quantitative studies on T cell diversity. IV. Mathematical analysis of multiple limiting populations of effector and suppressor T cells.

Limiting dilution (LD) analyses of polyclonally activated T cells yielded results suggesting the existence of multiple paired populations of effector and suppressor precursors for a number of different T cell functions and specificities analyzed. These populations occur at graded frequencies and suppression occurs within a pair but not between pairs. In this paper, we establish the mathematical basis for the interpretation of these multi-component limiting dilution results. First, we derive equations for a number of mathematical models and identify one model that both makes biological sense and can be used to reproduce experimental data. Second, within this model, we identify parameters such as the frequency of suppressive cells and the number of suppressive cells required for suppression. The results suggest that within each paired population, suppressor precursors are 20 times more frequent that effector precursors. Furthermore, a similar but variable excess of suppressor cells is required for suppression to become effective. Together with the high frequency (1/50-1/500) of most effector T cell precursors previously reported, the results suggest that up to 40% of the T cells can become involved in suppression of an antigen-specific effector T cell population. These studies may provide exact estimates for predictions to be tested in experiments on immune regulation.

Quantitative studies on T cell diversity. III. Limiting dilution analysis of precursor cells for T helper cells reactive to xenogeneic erythrocytes.

Splenic T cells exposed to concanavalin A (Con A), and subsequently to factors produced by rat spleen cells in response to Con A (Con A sup), acquire the ability to function as helper T (TH) cells in response to xenogeneic erythrocytes (RBC). Help is measured as the reconstitution of the plaque-forming cell response of a spleen cell population depleted of T cells by treatment with anti-Thy-1 serum and complement. We propose that precursor TH cells differentiate during the in vitro treatment into mature TH cells. As differentiation occurs under limiting dilution conditions, an estimation of the precursor frequency should in principle be possible. However, a single-hit Poisson distribution does not fit our data. Instead, we observe, dependent on the T cell concentration, three separate "peaks" of response. In many experiments, using sheep, horse, and chicken RBC as antigens, we reproducibly find these "peaks" at 40-190, 600-3,000, and 20,000-100,000 T cells, placed into limiting dilution cultures, respectively. By various experiments we can show that the helper activity is not due to passively transferred rat factors, but to the titrated cells themselves. The active cell is a T cell that appears to function in an antigen-specific way and to require direct cell contact to do so. It thus resembles the classical helper T cell. As we find precursor TH cells already at very low concentrations of T cells, we titrated the range between 0 and 100 T cells/well carefully. The bent shape of the titration curves does not always allow a statistically satisfying regression analysis, and we therefore cannot estimate precise precursor frequencies from every experiment. However, a common sense argument can be made that these frequencies must be on the order of 1/10-1/100 T cells. We propose that the limiting dilution curves obtained in this system most likely reflect fundamentally important cellular interactions that regulate immunological effector functions. We favor a concept of independently interacting sets of helper and suppressor T cells of various frequencies, but other models are possible.

Reducing postischemic damage by temporary modification of reperfusate calcium, potassium, pH, and osmolarity.

This study was designed to determine if ischemic damage could be reduced by modifying blood composition upon reperfusion. After control data had been obtained in seven dogs on prolonged cardiopulmonary bypass, 71 dogs underwent 1 hour of ischemic arrest with topical hypothermia (left ventricular temperature 16 degrees C). We measured left ventricular performance (isovolumetric function curves), compliance (intraventricular balloon), blood flow (microspheres), metabolism (oxygen consumption), and water content (wet/dry weights) before and 30 minutes after ischemia. The initial reperfusate was 500 cc of oxygenated blood given over a period of 5 minutes. Without temporary reperfusate modification, postischemic left ventricular performance was depressed 40% +/- 3%, compliance fell 50% +/- 12%, water content rose 2.5% +/- 0.1%, and left ventricular blood flow and oxygen uptake increased only minimally when cardiac work was increased (function curve). These deleterious changes were reduced significantly, but not prevented, by the following isolated reperfusate modifications: (1) lowering amount of ionic calcium available for cell entry, (2) raising pH to 7.8 to counteract acidosis, (3) raising potassium level to maintain arrest and reduce metabolic demands, and (4) increasing osmolarity (mannitol, 360 mOsm) to counteract edema. In contrast, by combining these modifications to achieve a hypocalcemic, hyperkalemic, alkalotic, and hyperosmolar blood perfusate, it was possible to attain 104% +/- 1% recovery of myocardial performance, 80% +/- 1% restoration of compliance, 60% less postischemic edema, and near-normal augmentation of left ventricular flow and oxygen uptake to meet increasing needs.

Superiority of blood cardioplegia over asanguinous cardioplegia--an experimental and clinical study.

This study compares experimental and clinical results using both an asanguinous and sanguinous vehicle for delivering the cardioplegic solution. Animals receiving blood cardioplegia had better left ventricular function after unclamping, significantly less post-ischemic myocardial edema, and a better ability to augment flow and oxygen consumption. Patients receiving blood cardioplegia had also better myocardial performance with high cardiac outputs and lower left atrial pressure, and showed less evidence of myocardial damage. We conclude that oxygenation of the cardioplegic solution provides superior myocardial protection to that seen when the same solution is used in an asanguinous vehicle.

Regional myocardial blood flow after hypothermic arrest and cardioplegia.

Malperfusion due to increased coronary vascular resistance is presumably one of the factors responsible for incomplete functional recovery of the heart after aortic cross-clamping. Myocaridal blood flow (MBF, radioactive microspheres) was measured before and after 60 min of hypothermic ischemia in 16 dogs on cardiopulmonary bypass. After ischemia the hearts were reperfused for 30 min. MBF was measured in the empty beating heart and in the isovolumetrically contracting ventricle loaded with enddiastolic volumes (EDV) of 10, 20 and 30 ml (intraventricular latex balloon).