Does a magnetic blanket induce changes in muscular blood flow, skin temperature and muscular tension in horses?

REASONS FOR PERFORMING STUDY: Static magnetic blankets are often claimed to induce increases in blood flow, reduce muscle tension and tenderness, and be beneficial in both prevention and treatment of musculoskeletal injuries in horses. However, there are no studies that confirm alleged beneficial effects of magnets on muscles of the back in healthy horses. OBJECTIVES: To investigate whether static magnets sewn into a blanket affect back muscle blood flow, skin temperature, mechanical nociceptive threshold (MNT) and behaviour in healthy horses. STUDY DESIGN: Prospective, randomised, blinded, placebo-controlled crossover study. METHODS: The following outcome measurements of the back of 10 healthy horses were performed; blood flow by photoplethysmography, skin temperature by use of thermistors in conjunction with digital infrared thermography, and MNTs by algometry. The horses' behaviour was filmed during the procedure and scored on an ethogram. Measurements were performed repeatedly for a 30 min baseline period. Thereafter a blanket with active, static magnets (900 gauss) or placebo magnets was placed on the horse and measurements were performed for a 60 min treatment period and a 30 min post treatment period. The study procedure was repeated on the consecutive day, when the horse received the alternative treatment. RESULTS: Blood flow in muscle, skin temperatures, MNTs and behavioural traits did not differ between active and placebo magnetic blankets. Skin temperature increased similarly during both active and placebo blanket treatment. CONCLUSIONS: In healthy horses, magnetic blankets did not induce additional significant effects on muscle blood flow, skin temperature, MNTs and behaviour when compared with nonmagnetic blankets.

Genetic analysis of insect bite hypersensitivity (summer eczema) in Icelandic horses.

There is a lack of knowledge about the genetic background of eczema due to insect bite hypersensitivity, also called summer eczema, in horses. The condition is known in several horse breeds and countries and it causes reduced welfare of the horse and economic losses to the owner. The aim of this study was to estimate genetic parameters for summer eczema in Swedish-born Icelandic horses. A questionnaire was sent to owners of horses sired by stallions with more than 50 offspring born in Sweden between 1991 and 2001. Variance components of summer eczema classified as healthy, mild, moderate or severe were estimated using the threshold methodology with sire models. In addition, summer eczema was analysed as a binary trait (healthy v. affected). The analyses included 1250 horses sired by 33 stallions. The prevalence of summer eczema was 8%, with a range of 0% to 30% in different paternal half-sib groups. Offspring of dams suffering from eczema had a higher risk of developing eczema. The heritability for severity of summer eczema was estimated at 0.3 (s.d. < 0.2) with a threshold sire model. In contrast to the age of the horse, different geographic areas and gender were significantly associated with severity of the eczema. We conclude that genetic selection could decrease the prevalence of summer eczema among Swedish-born Icelandic horses. The amount and quality of data are, however, crucial for the possibility to introduce a genetic evaluation of summer eczema. The symptoms should be classified in several classes according to severity, and this classification could be made by the horse owner.

Clinical signs and etiology of adverse reactions to procaine benzylpenicillin and sodium/potassium benzylpenicillin in horses.

Case reports of 59 horses reacting adversely to procaine benzylpenicillin or to sodium or potassium benzylpenicillin in Sweden in 2003-2005 were obtained through contacts with horse-owners. For the assessment of the reports, various parameters were evaluated, such as the times to the reactions, information on previous penicillin treatment, the clinical signs and the actions taken in the reacting horses. Among the reports, two horses had received sodium or potassium benzylpenicillin intravenously, whereas the remaining 57 horses had been treated with procaine benzylpenicillin intramuscularly. Allergy may underlie the adverse reactions in the horses given sodium and potassium benzylpenicillin, and in a few of the horses given procaine benzylpenicillin. However, in most horses in the latter group, the clinical signs may be due to the toxic effects of procaine. In these horses, the dominating clinical signs were locomotor and behavioral changes. Some risk factors may enhance the probability that horses react to procaine. One is repeated injections, which increase the likelihood of intravascular administration and also may increase the sensitivity to procaine due to neuronal sensitization (kindling). Procaine is rapidly hydrolyzed by plasma esterases to nontoxic metabolites. When high amounts of procaine enter the circulation, the hydrolyzing capacity may be exceeded and toxicity occurs. Analyses of plasma esterases from reacting horses showed lower activity than in nonreacting control horses. Low esterase activity may increase the possibility of procaine toxicity and constitute another risk factor.

Cetirizine in horses: pharmacokinetics and effect of ivermectin pretreatment.

The pharmacokinetics of the histamine H(1)-antagonist cetirizine and the effects of pretreatment with the antiparasitic macrocyclic lactone ivermectin on the pharmacokinetics of cetirizine were studied in horses. After oral administration of cetirizine at 0.2 mg/kg bw, the mean terminal half-life was 3.4 h (range 2.9-3.7 h) and the maximal plasma concentration 132 ng/mL (101-196 ng/mL). The time to reach maximal plasma concentration was 0.7 h (0.5-0.8 h). Ivermectin (0.2 mg/kg bw) given orally 1.5 h before cetirizine did not affect its pharmacokinetics. However, ivermectin pretreatment 12 h before cetirizine increased the area under the plasma concentration-time curve by 60%. The maximal plasma concentration, terminal half-life and mean residence time also increased significantly following the 12 h pretreatment. Ivermectin is an inhibitor of P-glycoprotein, which is a major drug efflux transporter in cellular membranes at various sites. The elevated plasma levels of cetirizine following the pretreatment with ivermectin may mainly be due to decreased renal secretion, related to inhibition of the P-glycoprotein in the proximal tubular cells of the kidney. The pharmacokinetic properties of cetirizine have characteristics which are suitable for an antihistamine, and this substance may be a useful drug in horses.

Fexofenadine in horses: pharmacokinetics, pharmacodynamics and effect of ivermectin pretreatment.

The pharmacokinetics and the effects on inhibition of histamine-induced cutaneous wheal formation of the histamine H1-antagonist fexofenadine were studied in horse. The effect of ivermectin pretreatment on the pharmacokinetics of fexofenadine was also examined. After intravenous infusion of fexofenadine at 0.7 mg/kg bw the mean terminal half-life was 2.4 h (range: 2.0-2.7 h), the apparent volume of distribution 0.8 L/kg (0.5-0.9 L/kg), and the total body clearance 0.8 L/h/kg (0.6-1.2 L/h/kg). After oral administration of fexofenadine at 10 mg/kg bw bioavailability was 2.6% (1.9-2.9%). Ivermectin pretreatment (0.2 mg/kg, p.o.) 12 h before oral fexofenadine decreased the bioavailability to 1.5% (1.4-2.1%). In addition, the area under the plasma concentration-time curve decreased 27%. Ivermectin did not affect the pharmacokinetics of i.v. administered fexofenadine. Ivermectin may influence fexofenadine absorption by interfering in intestinal efflux and influx pumps, such as P-glycoprotein and the organic anion transport polypeptide family. Oral and i.v. fexofenadine significantly decreased histamine-induced wheal formation, with a maximal duration of 6 h. A pharmacokinetic/pharmacodynamic link model indicated that fexofenadine in horse has antihistaminic effects at low plasma concentrations (EC50 = 16 ng/mL). However, oral treatments of horses with fexofenadine may not be suitable due to the low bioavailability.

Pharmacokinetics and pharmacodynamics of clemastine in healthy horses.

Clemastine is an H1 antagonist used in certain allergic disorders in humans and tentatively also in horses, although the pharmacology of the drug in this species has not yet been investigated. In the present study we determined basic pharmacokinetic parameters and compared the effect of the drug measured as inhibition of histamine-induced cutaneous wheal formation in six horses. The most prominent feature of drug disposition after intravenous dose of 50 microg/kg bw was a very rapid initial decline in plasma concentration, followed by a terminal phase with a half-life of 5.4 h. The volume of distribution was large, Vss = 3.8 L/kg, and the total body clearance 0.79 L/h kg. Notably, oral bioavailability was only 3.4%. There was a strong relationship between plasma concentrations and effect. The effect maximum (measured as reduction in histamine-induced cutaneous wheal formation) was 65% (compared with controls where saline was injected) and the effect duration after i.v. dose was approximately 5 h. The effect after oral dose of 200 microg/kg was minor. The results indicate that clemastine is not appropriate for oral administration to horses because of low bioavailability. When using repeated i.v. administration, the drug has to be administered at least three to four times daily to maintain therapeutic plasma concentrations because of the short half-life. However, if sufficient plasma concentrations are maintained the drug is efficacious in reducing histamine-induced wheal formations.

Prevalence and risk factors of bone spavin in Icelandic horses in Sweden: a radiographic field study.

The present field study was performed to estimate the prevalence of radiographic signs of bone spavin (RSBS) and to evaluate some possible risk factors for RSBS in the Icelandic horse in Sweden. The survey included horses from 11 farms involving 379 horses, comprising 238 geldings, 125 mares and 16 stallions. Age ranged from 0 to 19 years with a mean age of 8.1 years. Horses were radiographed with a dorsolateral-plantaromedial-oblique projection of both tarsi. Information about age, gender, origin, working intensity, number of gaits and age when saddle broken were obtained by interviewing the owners. Features of each horse's conformation were obtained by measurement or inspection. Radiographic signs of bone spavin were found in 23% (n = 88) of the horses. Horses younger than 5 years did not show any RSBS and the prevalence increased from zero to 33% between 4 and 8 years of age. Using a multivariate logistic regression model increasing age and sickle hock conformation were found to be significantly associated with increased risk of RSBS. Age when saddle broken in combination with height at the withers also influenced the multivariate logistic regression model significantly.

Relationship between hind limb lameness and radiographic signs of bone spavin in Icelandic horses in Sweden.

A field study was designed to estimate the prevalence of hind limb lameness in a population of Icelandic horses in Sweden. All available Icelandic horses at 11 different farms within 150 km from Uppsala were examined once during a study period of 13 months. The relationship between hind limb lameness and radiographic signs of bone spavin (RSBS) was investigated. The rate of agreement between the outcome after palpation, motion evaluation and flexion test and radiographic examination was established. A total of 379 horses including 238 geldings, 125 mares and 16 stallions, with mean age 8.1 years (range 1-19 years) were examined by palpation of the medial aspect of the tarsal joints, motion evaluation and flexion test of the hind limbs. One dorsolateral-plantaromedial oblique radiographic view was recorded from each tarsus. Palpable abnormalities were found in 118 horses (31%). Forty-three horses (12%) were lame at presentation, 94 horses (25%) were positive on flexion test and 88 horses (23%) had RSBS. There was a significant relationship between palpable abnormalities, lameness, positive flexion test and RSBS. The proportion of joints with RSBS increased with increasing severity of physical findings. The proportion of lame limbs increased with increasing radiographic changes. The presence of palpable findings or positive flexion test was not influenced by the grading of radiographic changes. Using palpation, motion evaluation, and flexion test 75% of the horses with RSBS were identified. The rate of agreement between the clinical examination and the radiographic examination was 73%.

Generation of in vitro natural cytotoxicity of horse lymphocytes against sarcoid-derived tumor cells not expressing major histocompatibility complex antigens.

OBJECTIVE: To analyze in vitro lymphocyte-mediated immune responses of horses with sarcoids against allogeneic sarcoid cells containing endogenous retrovirus but not expressing major histocompatibility complex antigens. DESIGN: Lymphocyte-mediated immune reactions were assessed by means of proliferative responses in mixed lymphocyte tumor cell culture (MLTC) assay and lymphocyte-mediated cytotoxicity against various equine target cells. ANIMALS: 12 horses with sarcoid tumors and 15 control horses. PROCEDURE: Blood lymphocytes were cocultured in MLTC with allogeneic sarcoid cells (Mc-1, BayMc-1), equine testis cells, or normal equine dermal fibroblasts. Lymphocytes were assayed for proliferative responses by [3H]thymidine uptake and for cytotoxicity against the same targets by 51Cr release assay. The lymphocyte populations were analyzed for some common surface markers. RESULTS: Lymphocytes from horses with sarcoids exerted an anamnestic proliferative response in MLTC against Mc-1 cells, but this procedure never generated cytotoxic lymphocytes. However, lymphocytes from all horses cultured in medium with 10% allogeneic serum only had selective. natural cytotoxicity against Mc-1 that was generated without DNA synthesis. Approximately 80% of the lymphocytes disappeared during culture; however the remaining population of small, viable lymphocytes indicated a decrease of CD4+ T lymphocytes, but numbers of T cells with receptors for Helix pomatia A hemagglutinin were unaffected. Few lymphocytes had Fc-receptors for IgG, were complement-reactive positive cells or were B cells expressing surface immunoglobulin. CONCLUSIONS: Results may indicate a natural defense system, which preferentially recognizes and lyses tumor cells that are deficient in surface expression of major histocompatibility complex antigens, without intervention of conventional T-cell receptors or antibodies.

Equine sarcoids. A clinical and epidemiological study in relation to equine leucocyte antigens (ELA).

Associations between clinical parameters of sarcoids and the equine leucocyte antigen system (ELA) were analysed for 120 Swedish horses. Median age of affected horses was 5.2 years, and the majority presented with solitary tumors between 2 and 5 cm in diameter and ventral abdomen was a predilection site. Clinical signs first appeared at a median age of 3.5 years, and sarcoids at different locations first appeared at different ages. Lesions at different sites differed in size, and multiple tumors, early onset, long duration, and older age all had an association with large size. Clinical manifestations of sarcoids and the association between certain ELA-specificities and early onset (A5) and increased recurrence rates after surgery (W13), in addition to increased prevalence (A3W13), strengthen further that some horses are inherently predisposed to sarcoid growth. Unassociated with any clinical parameters, one third of the untreated horses became free of sarcoids due to "spontaneous" regression, perhaps as a result of immune responses against the tumors. Seventy percent of the horses were treated (mostly by excision), and large size was the main parameter promoting treatment. Excision had no significant effect on possibly remaining sarcoids. Recurrence rate after first treatment was about 35%, with the majority of tumors recurring within 4 months. Early onset, long duration, large size, and localization to distal limbs all appeared to increase risk of recurrence. Early treatment, performed under general anesthesia in recumbency which permits wide excision and measures to avoid autoinoculation, significantly reduced recurrence rates.

Association between equine leucocyte antigens (ELA) and equine sarcoid tumors in the population of Swedish halfbreds and some of their families.

The distribution of equine leucocyte antigens (ELA) in Swedish Halfbreds affected by sarcoid tumors was determined and compared with that of control horses of the same breed. ELA-haplotype A3W13 appeared more frequently in affected horses, resulting in a chi 2 value of 4.45 (P = 0.034) for A3 and 9.05 (P = 0.0026) for W13, respectively. The relative risk factor (RR) could be estimated to 2.13 and 3.00 for A3 and W13, respectively. The etiology fraction (EF) was calculated to 28% and 37% for A3 and W13, respectively. Thus, in the population of Swedish Halfbreds approximately 40% (at least) of the disease appeared to be associated with the genetic background of the affected horse. Family studies established that ELA are codominantly expressed and inherited as simple Mendelian traits and that sarcoids among offspring are significantly associated with one of the parental haplotypes (P = 0.00942). This parental haplotype does not always include A3W13. These results confirm and extend previous results from other breeds and strongly suggest the existence of a predisposition for sarcoids among horses, that is due to an autosomal, dominant, ELA-linked gene with incomplete penetrance. In extension, this indicates a multifactorial etiology of equine sarcoids (additional non-MHC gene(s) and/or environmental factors).

Allergic dermatitis (sweet itch) of Icelandic horses in Sweden: an epidemiological study.

A survey of allergic dermatitis (sweet itch) in Sweden contained information on 441 Icelandic horses. Results of a questionnaire indicated that approximately 15 per cent of the country's Icelandic horses suffered from the disease. The prevalence of allergic dermatitis was significantly higher among horses imported from Iceland (26.2 per cent) compared to that of Swedish-born animals (6.7 per cent). In addition, horses born in Iceland were significantly more severely affected than horses born in Sweden. The risk of allergic dermatitis in Sweden appeared to be more than six times higher for horses exported from Iceland to Sweden relative to that of horses originally born in Sweden. The prevalence of disease for horses of seven years or older was 30 per cent for Icelandic-born individuals as compared to 7.3 per cent for horses born in Sweden. Similarly, the risk of allergic dermatitis in Sweden for horses of seven years or older appeared to be nearly 10 times higher for horses imported from Iceland relative to that of horses born in Sweden. Allergic dermatitis usually appeared during the third grazing season for imported horses and during the fourth season for horses born in Sweden. Furthermore, the course of the disease tended to become worse with time. Analysis of the prevalence of allergic dermatitis relative to gender revealed no significant differences. Certain geographical variations in the prevalence of the disease was also found.

Natural cytotoxicity of human lymphocytes against equine target cells in vitro.

Human lymphocytes displayed a frequent natural cytotoxicity (NK) in vitro against normal equine dermal fibroblasts (ED) and against equine tumour cells of a virus-containing cell line (Mc-1). Similarly, human normal sera contained antibodies that induced antibody-dependent cellular cytotoxicity (ADCC) by normal human lymphocytes against the same target cells. Both NK and ADCC varied for different donors. For individual donors, however, cytotoxicity against the two target cells was significantly correlated both in NK and ADCC. For ED there was also a significant correlation between ADCC and NK activity. Both NK and ADCC showed some selectivity as assessed by cold target cell inhibition. Inhibition studies with Fab fragments of anti-human IgG established the involvement of immunoglobulins in the NK reaction. In this context, a marked and mainly immunoglobulin-dependent increase in both NK and ADCC activity against Mc-1 was observed in a laboratory worker frequently exposed to the target cells. The results indicate that variations of natural cytotoxicity in individual donors may sometimes be an indication of an ongoing spontaneous sensitization.

A new surface marker on equine peripheral blood lymphocytes. I. Subpopulations of lymphocytes with receptors for Helix pomatia A hemagglutinin (HP).

Untreated and neuraminidase-treated equine peripheral blood lymphocytes were analysed for binding of the A hemagglutinin of the snail Helix pomatia (HP). For optimal staining by direct immunofluorescence, the concentration of neuraminidase had to be increased as compared to that needed for other species. Moreover, higher concentrations of HP were required for optimal staining of equine lymphocytes as compared to lymphocytes from other species. Even so, the maximal number of equine lymphocytes exhibiting positive staining was only about 20%. No, or very few, HP-positive lymphocytes were seen when neuraminidase treatment was omitted. However, when the more sensitive method of indirect immunofluorescence was used, approximately 60% of the lymphocytes were HP positive without prior treatment with this enzyme. Neuraminidase treatment significantly increased this figure to about 75%. In all instances, HP binding was specific since it was inhibited by the competitive sugar hapten N-acetyl-D-galactosamine (D-GalNAc) while addition of D-glucose (D-Glc) gave no inhibition. HP binding to neuraminidase-treated lymphocytes was also investigated quantitatively by means of 125I-labeled HP. The number of HP molecules bound per HP-positive cell was approximately 3 X 10(5) and the apparent association constant for the binding of HP to its cellular receptors was approximately 8 X 10(7) 1/mol. No binding of HP to untreated lymphocytes could be recorded in these experiments.

A new surface marker on equine peripheral blood lymphocytes. II. Characterization and separation of purified blood lymphocytes with receptors for Helix pomatia A hemagglutinin (HP).

In a preceding report we have shown that the lectin Helix pomatia A hemagglutinin (HP) binds to two subpopulations of neuraminidase-treated equine peripheral blood lymphocytes (PBL), constituting about 20% and 75% of PBL, respectively. The aim of the present study was to further characterize these HP+ cells in regard to other surface markers such as receptors for guinea pig erythrocytes (GPR+ cells), membrane-bound immunoglobulins (sIg+ cells), receptors for activated complement (C3R+ cells) and receptors for IgG (Fc alpha R+ cells). This was done by double marker analysis and by lymphocyte fractionation either on columns charged with HP coupled to Sepharose beads or by rosetting with guinea pig erythrocytes. The fractions were also analysed for their proliferative response in the mixed lymphocyte tumor cell interaction (MLTC) assay and to the mitogens leucoagglutinin (La) and concanavalin A (Con A). The results revealed that the majority of GPR+ cells also expressed high avidity receptors for HP, as defined by means of direct immunofluorescence. These cells constituted a subpopulation of GPR+/HP+ cells T cells comprising approximately 20% of PBL. In contrast, about 75% of the HP+ cells in indirect immunofluorescence were GPR-. The fractionation experiments showed that HP+ and GPR+ cells were probably not B cells since they were sIg-. The C3R+ and Fc alpha R+ lymphocytes were heterogeneous in regard to HP receptors but the majority of these cells was also found in the fractions depleted of HP+ and GPR+ lymphocytes. The fractions eluted from HP columns gave a strong proliferative response in MLTC, whereas fractions depleted of HP+ cells responded poorly. However, in contrast to the GPR+-depleted fractions, those enriched in GPR+ lymphocytes responded poorly to the T cell mitogens La and Con A. The mitogenic response of the HP-column fractions to La and to Con A was variable. The results are discussed in relation to HP being a surface marker for a heterogeneous population of equine T cells.

Cell-mediated immunity in horses with sarcoid tumors against sarcoid cells in vitro.

Cell-mediated immunity in horses with sarcoid tumor against sarcoid antigens was studied in vitro by means of mixed lymphocyte tumor cell culture assay and lymphocyte-mediated cytotoxicity of 52Cr-labeled target cells. When Mc-1 sarcoid cells were used as stimulatory cells for peripheral blood lymphocytes in the mixed lymphocyte tumor cell assay, a clear difference in the kinetics of the generated lymphocytic proliferative response could be detected between sarcoid and control horses. With sarcoid horses, their proliferative maximum was reached 3 days earlier than that of the control horses, and at this time their proliferative activity was significantly increased over that of control horses. When normal allogeneic fibroblasts were used as stimulatory cells, no such difference between sarcoid and control horses could be seen. The cellular cytotoxicity of peripheral blood lymphocytes from sarcoid and control horses against Mc-1 cells or normal allogeneic fibroblast targets was very low. However, the mean cytotoxicity against Mc-1 was slightly increased for sarcoid horses as compared with that of control horses. In contrast, the cytotoxicity against allogeneic fibroblasts was slightly lower for sarcoid than for control horses. In contrast, the cytotoxicity against allogeneic fibroblasts was slightly lower for sarcoid than for control horses. Furthermore, it was shown that sarcoid horses, but not control horses, had a slightly but consistently increased cytotoxicity against Mc-1 cells as compared with that against normal allogeneic fibroblasts.

A simple Beckman DU accessory for location of proteins separated by isoelectric focusing in granulated gel layers: isolation of human serum very low density apolipoproteins C-II and C-III-1.

A simple, low-cost gel layer scanner has been developed for the Beckman DU spectrophotometer. The gel scanner makes it possible to localize zones precisely after preparative isoelectric focusing of proteins in 2-mm thick Sephadex G-200 layers. Using the scanner after isoelectric separation of a mixture of human serum apolipoproteins C-III-1 and C-II, pure proteis could easily be obtained.