Intradermal challenge of Icelandic horses with extracts of four species of the genus Culicoides

Twenty-three Icelandic horses were challenged with extracts of four species of biting midges: Culicoides pulicaris, C chiopterus, C obsoletus and C impunctatus. Fourteen of the tested horses were affected with summer eczema. The horses were challenged intradermally with 0.1 ml of whole-body extracts of midges at a concentration of 0.01 or 0.005 per cent weight/volume. The skin reactions were measured after 30 minutes, 60 or 180 minutes and four, 24 and 48 hours after injection. Antigen titration showed that the reaction was dependent on the antigen concentration. Eight of nine unaffected horses failed to respond to any of the four antigens; the remaining animal responding to two of the four antigens. Ten of the 14 affected horses responded to at least three of the four antigens, while two of the animals in this group failed to respond to any. The mean responses to C chiopterus, C obsoletus and C impunctatus, read after 30 minutes, 60 minutes and four hours were significantly higher in the affected horses than in the unaffected horses. A significant difference was also found in the mean response to C chiopterus and C impunctatus, read after 24 hours.     

Intradermal skin tests in equine dermatology: a study of 83 horses

Allergic diseases are often diagnosed clinically in the horse without performing diagnostic tests. The purpose of this work was to contribute to the validation of intradermal skin tests in the horse. Eighty-three horses, 14 showing skin or respiratory signs of supposed allergic origin, were subjected to an intradermal skin test using 6 different allergens, positive and negative controls. The tests were read for all animals after 20 min, and for 29 horses after 1 and 4 h. Additionally, 19 horses were tested a few months apart. The comparison after 20 min of the cutaneous reactions to allergens and to positive and negative controls allowed us to propose a threshold of positivity; skin reactions with a diameter above or equal to 15 mm, or with at least a 13 mm diameter and a skin thickness similar to the positive control. There was a marked difference between the healthy group and the allergic group for Culex pipiens and Dermatophagoides farinae, although positive reactions were not rare in the healthy group. Tabanus sp. gave positive reactions in both healthy and allergic animals. There was no significant variation in the reactions observed after 20 min and after 60 min. After 4 h, the progression of the reactions was highly variable and negative controls showed a certain number of positive reactions, which negated their reliability at the point in time, and made it difficult to interpret the other allergens. Finally, the repeatability of intradermal skin tests reactions after 20 min was poor, and probably related to the influence of seasonality for some of the allergens. Further studies are required, notably with others allergens, to interpret intradermal skin tests' responses in clinical practice.     

Intradermal skin testing in Hispaniolan parrots (Amazona ventralis)

The purpose of this study was to develop and standardize a protocol for intradermal skin testing in birds. Forty clinically normal Hispaniolan Amazon parrots were anaesthetized and tested by intradermal injection with 0.02 mL of phosphate-buffered saline, histamine phosphate, compound 48/80, codeine phosphate, deionized water, antiavian IgG and rabbit serum. Injection sites were evaluated at 5, 10 and 15 min, 4–6, 24 and 48 h following injection using callipers to measure the diameter of the wheals. A second intradermal skin test was repeated in 20 birds with 0.03 mL of saline, compound 48/80 and codeine phosphate. This study provides the basis for an appropriate protocol for intradermal skin testing in parrots, including recommended site (proventer region), volume of injection (0.02 mL), negative control (saline), positive control (codeine phosphate 1 : 100 000 w/v) and optimum reading time (5 min). Further study to establish appropriate dosages for test antigen will be required.     

Intradermal testing of horses with chronic obstructive pulmonary disease and recurrent urticaria.

Six horses with chronic obstructive pulmonary disease (COPD) and 8 horses with recurrent urticaria were skin tested with 67 extracts from 58 allergens, including pollens, epidermals, cultivated farm plants, dusts, molds, and insects. Reactions were evaluated 3 times over a 24-hour period immediately after the injections. Results were compared with those obtained from 11 clinically normal horses. All horses had positive skin test reactions. Significant differences was evident between horses with COPD and clinically normal horses for only 3.0% of the possible extract reactions, and between horses with urticaria and clinically normal horses for only 4.5% of the possible extract reactions. Horses with COPD or urticaria had greater total percentage of allergen extract reactions than did clinically normal horses. Positive reactions were observed at all 3 evaluation periods, and late-onset reactions were not always preceded by positive reaction at earlier periods. All horses with COPD or urticaria had at least 1 skin test reaction that exceeded the mean +/- 2 SD, as calculated for each of the 67 extracts for the group of clinically normal horses.     

Intradermal testing in healthy horses and horses with chronic obstructive pulmonary disease, recurrent urticaria, or allergic dermatitis.

OBJECTIVE: To compare responses to a variety of intradermally injected allergens among healthy horses and horses with chronic obstructive pulmonary disease (COPD), recurrent urticaria (RU), and atopic dermatitis-insect hypersensitivity (allergic dermatitis [AD]). DESIGN: Case-control study. ANIMALS: 86 horses. PROCEDURE: Results of intradermal testing for horses with COPD, RU, or AD were compared with results for healthy horses. RESULTS: Compared with healthy horses, horses with COPD, RU, and AD were significantly more likely to have positive (> or = 3+) reactions to intradermal allergens (molds, weeds, trees, grasses-crops, and insects) 30 minutes (immediate reaction), 4 hours (late-phase reactions), and 24 hours (delayed-phase reactions) after exposure. In addition, diseased horses reacted to a significantly higher number of allergens in each allergen group than did healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE: Reactions to individual allergens should not be used to determine that horses have hypersensitivity. Overall patterns of reactivity to intradermal allergens may be helpful in management when used in conjunction with a compatible history and evidence of potential exposure to allergens in horses with conditions associated with hypersensitivity to environmental allergens.     

Application of the ASVCP guidelines for the establishment of haematologic and biochemical reference intervals in Icelandic horses in Austria

Background

Despite the increasing popularity of Icelandic horses, published reference intervals (RIs) in this breed are rare. Due to their isolation and their small gene pool, alterations in some variables are likely and some possible breed-specific peculiarities have been described. The purpose of the present study was the establishment of comprehensive RIs in Icelandic horses according to recently published guidelines.In a prospective observational study, blood samples were collected from the jugular vein of 142 Icelandic horses into EDTA and serum tubes. Reference intervals were established for haematologic and biochemical analytes on the Advia 2120i™ and the Dimension ExL™ by established methods. RIs were defined as central 95 % intervals bounded by the 2.5th and 97.5th percentiles with their 90 % confidence intervals, calculated according to recently published ASVCP guidelines. An inhouse-developed quality control system using observed total allowable error was used for the surveillance of the internal quality control preceding the measurements.

Results

The RIs were as follows: haematocrit: 0.29–0.39, RBC: 5.79–8.63 T/l, haemoglobin: 102.0–142.3 g/l, MCV: 42–51 fl, platelets: 146–263 G/l, WBC: 4.13–8.57 G/l, segs: 1.98–4.73 G/l, lymphocytes: 1.25–3.49 G/l, monocytes: 0.06–0.31 G/l, eosinophils: 0.04–0.50 G/l, glucose: 4.0–5.7 mmol/l, urea: 3.2–6.4 mmol/l, creatinine: 79.6–141.4 μmol/l, total protein: 54.4–72.9 g/l, albumin: 27.7–36.8 g/l, total bilirubin: 8.1–21.1 μmol/l, triglycerides: 0.03–0.44 mmol/l, cholesterol: 1.75–2.90 mmol/l, ALP: 1.35–3.55 μkat/l, AST: 4.52–8.80 μkat/l, GLDH: 0.0–0.18 μkat/l, GGT: 0.11–0.39 μkat/l, CK: 2.53–6.52 μkat/l, LDH: 3.32–7.95 μkat/l, iron: 16.4–39.9 μmol/l, calcium: 2.69–3.19 mmol/l, phosphate: 0.5–1.3 mmol/l, magnesium: 0.6–0.9 mmol/l, sodium: 134–141 mmol/l, potassium: 3.6–4.7 mmol/l, chloride: 100–105 mmol/l.

Conclusions

Reference intervals of several haematologic and biochemical analytes differed from the transferred historical reference intervals applied to equine samples in the authors’ laboratory. These might be of clinical importance in some analytes such as creatine kinase.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-015-0120-4) contains supplementary material, which is available to authorized users.     

Nutritional value of timothy haylage in Icelandic horses

Four mature Icelandic geldings were used in a balance trial to investigate the effect of stage of maturity of timothy (Pleum pratence L.) haylage fed at maintenance level on coefficients of total tract apparent digestibility (CTTAD), and nitrogen and energy metabolism in a 4 × 4 change-over arrangement of treatments. The content of fibre increased and crude protein content decreased with advancing stage of maturity. Very high CTTAD values were found for DM, OM, CP, CF, NDF and ADF in the first two cuts. Overall, there was a negative relationship between stage of maturity (p < 0.05) and the CTTAD of all measured dietary components. Nitrogen (N) intake, N losses in urine and N balance were affected (p < 0.05) by stage of maturity, while the N losses in the faeces were unaffected. There was a strong correlation (r² = 0.95) between N balance and water intake. Gross energy intake and losses in urine (as % of digestible energy intake) were unaffected by stage of maturity, while the energy balance was affected (p < 0.05). The digestible energy content of the timothy haylage studied ranged from 14.1 to 9.0 MJ/kg DM.

In conclusion, early-cut timothy haylage (stem elongation to flowering) has the energy content required to form the basis of diets for high-performing horses. Late-cut timothy haylage (seeding to late seeding), on the other hand, can be used to fulfil the energy and protein needs of maintenance-fed Icelandic horses.     

Nutritional value of mixed grass haylage in Icelandic horses

The influence of cutting time of mixed grass haylages on energy and nitrogen (N) metabolism, and the coefficients of total tract apparent digestibility (CTTAD) were studied in four mature Icelandic geldings by performing total collection of faeces and urine. The experiment was arranged as 4 × 4 balanced Latin square. The four haylages used were harvested at different times and were considered to be representative in botanical composition for mixed grass haylages in Iceland.     

Risk factors for faecal sand excretion in Icelandic horses

REASONS FOR PERFORMING STUDY: Sandy soil is often mentioned as a risk factor in the development of sand-related gastrointestinal disease (SGID) in the horse. There are other variables, but few studies confirm any of these. OBJECTIVE: To investigate soil type, pasture quality, feeding practice in the paddock, age, sex and body condition score as risk factors for sand intake in the horse. METHODS: Faeces were collected from 211 Icelandic horses on 19 different studs in Denmark together with soil samples and other potential risk factors. Sand content in faeces determined by a sand sedimentation test was interpreted as evidence of sand intake. Soil types were identified by soil analysis and significance of the data was tested using logistic analysis. RESULTS: Of horses included in the study, 56.4% showed sand in the faeces and 5.7% had more than 5 mm sand as quantified by the rectal sleeve sedimentation test. Soil type had no significant effect when tested as main effect, but there was interaction between soil type and pasture quality. Significant interactions were also found between paddock feeding practice and pasture quality. CONCLUSION: To evaluate the risk of sand intake it is important to consider 3 variables: soil type, pasture quality and feeding practice. Pasture quality was identified as a risk factor of both short and long grass in combination with sandy soil, while clay soil had the lowest risk in these combinations. Feeding practice in the paddock revealed feeding directly on the ground to be a risk factor when there was short (1-5 cm) or no grass. Also, no feeding outdoors increased the risk on pastures with short grass, while this had no effect in paddocks with no grass. More than 50% of all horses investigated in this study had sand in the faeces. POTENTIAL RELEVANCE: The identification of risk factors is an important step towards prevention of SGID. Further research is necessary to determine why some horses exhibit more than 5 mm sand in the sedimentation test and whether this is correlated with geophagic behaviour.     

Genetic analysis of competition data on Icelandic horses

In a study of the possibility of using competition data in the genetic evaluation of Icelandic horses, data from competitions held in Iceland between 1999 and 2004 and in Sweden between 1998 and 2004 were analyzed to estimate the genetic parameters of competition performance traits. The data-sets from both countries included 18 982 records of 3790 horses in 379 different events. Two types of competition were included: sport competitions and gæðinga competitions performed on oval tracks and on a straight track. Each type of competition involves several disciplines in which the horses are ridden in the various gaits. The traits analyzed were two different measures of four-gait, five-gait and tölt, and one pace trait. In both the four-gait and five-gait tests the gaits walk, trot, tölt and gallop are exhibited. In the five-gait test, pace is also exhibited. The traits tölt and pace are performances of these single gaits. Highly correlated and similar traits were combined, and three new traits relating to tölt, four-gait and five-gait were formed. No large differences in means or standard deviations of traits were found between countries. All traits were approximately normally distributed. Genetic parameters were estimated using linear animal models including the fixed effects of sex, age and event for all traits, and the level of discipline was included for some traits. Random permanent environmental effects were also included. Estimated heritabilities were moderate to high, ranging from 0.18 to 0.21 for sport-competition traits, from 0.33 to 0.35 for gæðinga-competition traits and from 0.19 to 0.22 for combined traits. Estimated genetic correlations between different sport-competition traits varied from 0.63 to 0.96, and between the two gæðinga-competition traits it was estimated at 0.43. Genetic correlations between sport- and gæðinga-competition traits ranged from − 0.42 to 1.00. It was concluded that competition traits are suitable to include in genetic evaluations.     

Risk factors of colic in horses in Austria

A hospital‐based case study was conducted at the University of Veterinary Medicine Vienna for a 1‐year period. The purpose of this study was to determine possible alterable and non‐alterable risk factors of equine colic in Austria. The investigated parameters were obtained from hospital medical records (individual factors, duty of the horse, deworming, change in diet and water intake), from questionnaires (feed intake, watering types, housing and pasture practices) and from http://www.orf.at (weather‐related factors). Moreover, 221 collected feed samples were investigated through hygienic quality sensory evaluation and mould, yeast and bacteria presumptive samples were confirmed by microbiological investigation. Variables that were observed from the hospital medical records and found to be significantly associated with colic in a univariate analysis were included in multivariate analysis and the only remaining risk factor was decreased water consumption (p < 0.001, OR = 5.025). Consequently, a total of 2743 horses and 366 cases of colic were observed during the study period. The risk factors for colic in Austria were identified for the first time in this study and these increased risk factors were decreased water consumption, high amount of concentrate intake (p = 0.037), low hygienic quality of hay (p = 0.027) and high temperature on the arrival date (p = 0.003). Results suggest that the occurence of colic may not stop, but may decrease with better feed management practices in Austria.     

Genetic analysis of 'breeding field test status' in Icelandic horses

Genetic evaluation of Icelandic horses is currently based on results from breeding field tests where riding ability and conformation of the horses are evaluated over the course of 1-2 days. Only a small part of registered horses attend these field tests, and it can be assumed that these are not a random sample of the population. In this study, the trait test status was introduced, describing whether a horse was assessed in a breeding field test. This trait was analysed to find out whether it has a genetic variation and how it correlates genetically to other traits in the breeding goal. Breeding field test data included 39,443 mares born in Iceland in 1990-2001, of which 7431 were assessed in the period 1994-2007. The trait was defined in relation to age, gender and stud of horses. Variance and covariance components were estimated using the Markov Chain Monte Carlo method by applying the Gibbs sampler procedure in the DMU program. Three multivariate analyses were performed where the test status trait was analysed with breeding field test traits. Animal models and sire models were applied. Based on estimated heritabilities (0.51-0.67) and genetic correlations (0.00-0.87), the test status trait showed significant genetic variation and was strongly correlated to some traits. The test status trait reflects preselection in the breeding field test traits and should be included in the genetic evaluation to enhance the procedure, reduce selection bias and increase accuracy of the estimation.     

Distribution of leucocyte antigens in Icelandic horses affected with summer eczema compared to non-affected horses

Three hundred and three horses, exported from Iceland to Norway, Sweden, Denmark, Switzerland or Germany were tested for their distribution of leucocyte antigens. One hundred and thirty-six horses were affected with summer eczema. The panel of sera recognised the internationally accepted ELA-specificities A 1 to A10, and the nine work shop specificities W 11 to W 15 and W 18 to W 21. Also, some local specificities, characterised in Switzerland (Be I, Be III, Be 8, Be 25, Be 26, Be 27), and two non major histocompatibility complex (MHC)-linked antigens (Ely 1:1, Ely 2) were included. Only one antigen, Be 8, gave a statistically significant difference in distribution between the two populations: Relative risk = 2.5, x2 = 10.11, corrected P less than 0.01.     

Culling rate of Icelandic horses due to bone spavin

A survival analysis was used to compare the culling rate of Icelandic horses due to the presence of radiographic and clinical signs of bone spavin. A follow-up study of 508 horses from a survey five years earlier was performed. In the original survey 46% of the horses had radiographic signs of bone spavin (RS) and/or lameness after flexion test of the tarsus. The horse owners were interviewed by telephone. The owners were asked if the horses were still used for riding and if not, they were regarded as culled. The owners were then asked when and why the horses were culled. During the 5 years, 98 horses had been culled, 151 had been withdrawn (sold or selected for breeding) and 259 were still used for riding. Hind limb lameness (HLL) was the most common reason for culling (n = 42). The rate of culling was low up to the age of II years, when it rose to 0.05 for horses with RS. The risk ratio for culling was twice as high for horses with RS compared with horses without RS and 5.5 times higher for culling because of HLL. The risk of culling (prognostic value) was highest for the combination of RS with lameness after flexion test, next highest for RS and lowest for lameness after flexion test as the only finding. It was concluded that bone spavin affects the duration of use of Icelandic horses and is the most common cause of culling due to disease of riding horses in the age range of 7-17 years.     

An epidemiological study of summer eczema in Icelandic horses in Norway

A survey of summer eczema was conducted on 391 Icelandic horses in Norway. The study showed a prevalence of summer eczema of 17.6 per cent in the horses investigated. Icelandic horses born in Norway where shown to be less affected with summer eczema than imported horses, 8.2 per cent and 26.9 per cent respectively (P less than 0.001). There was no difference in the prevalence of summer eczema relative to gender or colour of the horse. The number of horses affected with summer eczema rose with increasing age and the number of years which had elapsed since importation. On average, Icelandic horses born in Norway developed summer eczema at the age of 5.3 years. The onset of disease for imported horses was, on average, 4.1 years after importation. The study also revealed that a significantly greater number of horses imported from Iceland to Norway during the period October to April, which has a minimal or low activity of insects (32.3 per cent), had developed summer eczema, compared with horses imported during the period May to September, when insects are more active (16.7 per cent), (P = 0.044). Horses with summer eczema were shown to be significantly more affected by respiratory disease than non-affected horses (P = 0.0086). The most common sites and clinical signs of summer eczema are also described.     

Influence of subclinical pulmonary findings on cardiac parameters in Icelandic horses

In the present study we examined, if in Icelandic horses an increase in heart and/ or breathing rate is physiological and breed dependend or a sign of a pulmonary or cardiac disease. Therefore we examined 37 Icelandic horses with the prereport of being healthy. During clinical lung examination four horses showed symptoms of a pulmonary disease like increased breathing rate and enforced breathing at rest. These horses were excluded from the study. The other 33 horses were clinically normal. 17 of these horses were unridden (untrained) and 16 horses were regularly worked (trained). After clinical examination in all horses analysis of arterial blood gas, endoscopy with tracheo- bronchial secret analysis and radiographic examination of the lung were carried out. Additionally electro- and echocardiographic examinations and standardised exercise tests with determination of heart and breathing rate as well as plasma lactate values were performed in all horses. During electro- and echocardiographic examination no pathological findings were observed. In total 22 of the 33 horses showed abnormal lung findings. Seven horses had mild signs of RAO and 15 horses had mild signs of interstitial bronchitis. Three horses had additional pulmonary haemorrhage. Eleven out of the 33 horses showed no abnormal lung findings. The breathing rate at rest differed not significantly between horses with (21 +/- 1/min) or without (23 +/- 2/min) pulmonary findings. The heart rate also did not differ significantly between horses with (39 +/- 1/min) or without (42 +/- 1/min) pulmonary findings. In contrast to this the trained Icelandic horses with abnormal pulmonary findings had significantly higher heart rates (p = 0.01) and significantly lower breathing rates (p = 0.009) compared to those without abnormal pulmonary findings. During echocardiography Icelandic horses with abnormal pulmonary findings had significantly larger left atrial diameter (without abnormal pulmonary findings: 82 +/- 7 mm, with abnormal pulmonary findings: 90 +/- 8 mm, p = 0.02). Compared to the untrained Icelandic horses (5.4 +/- 2 mmol/l) the trained horses showed significantly lower plasma lactate values (3.1 +/- 2 mmol/l, p = 0.001) immediately after exercise. After exercise the icelandic horses with abnormal pulmonary findings had significantly higher breathing rates (p < 0.05) and longer recovery periods (30 minutes) than horses without abnormal respiratory findings (15 minutes). Recovery of heart rate after exercise showed no differences between groups.     

Zinc and copper plasma levels in Icelandic horses with Culicoides hypersensitivity

Zinc concentration has been shown to have a potent immunomodulatory capacity, particularly influencing T helper cell organisation and cytokine secretion. Culicoides hypersensitivity (CHS) in horses resembles the early and late phase of type I hypersensitive reactions in man, characterised by a shift from T helper cell subtype 1 to T helper cell subtype 2 cytokine profile. In this pilot study, zinc and copper levels were measured in the plasma of 48 CHS-affected and 56 healthy Icelandic horses age 4-25 years (mean approximately 11 years) kept on 7 farms. Affected horses were divided into 3 groups according to the severity of disease. Time of blood collection and feeding management was constant. No differences in zinc or copper plasma levels and plasma copper/zinc ratio were determined among CHS horses and controls by univariate analysis of variance. Therefore, the most significant influences on zinc and copper plasma levels were affected by the location of housing. However, Spearman correlation showed a negative coefficient between the plasma zinc concentration and the severity of CHS (r = -0.31). Due to a probability value of P = 0.002 the null hypothesis r = 0 is rejected, although only 9% of the total variation of plasma zinc is presently explained by its relationship to CHS. In contrast, the Spearman correlation coefficient between plasma copper levels and severity of CHS was not significant (r = -0.14; P = 0.16). The minor deviations in plasma zinc concentrations in association with the severity of CHS may be real or due to neurohumoral or cytokine-mediated mechanisms, but appear too minimal to be relevant.     

Variations in the concentration of zinc in the blood of Icelandic horses

The effect of factors including the horses' farm environment, their sex and age and whether they suffered from summer seasonal recurrent dermatitis (sweet itch) on the concentrations of zinc in the plasma, whole blood and blood cells of 104 Icelandic horses was investigated. Its concentration in plasma varied significantly between farms (P<0.01), but its concentration in blood and blood cells was not influenced by any of the variables. The concentration of zinc in the blood cells was 10.5 times greater than in plasma, but its concentration in plasma was not correlated with its concentration in whole blood or blood cells owing to the variability in the proportion of whole blood zinc present in plasma (relative plasma zinc), which ranged between 9 and 24 per cent. This variability was significantly influenced by a three-way interaction between farm, sex and sweet itch (P<0.05). Relative plasma zinc was positively correlated with absolute plasma zinc (r=0.78, P<0.001) and negatively correlated with whole blood and blood cellular zinc (r=-0.58, r=-0.71, P<0.001).