Exploring the genetics underpinning dynamic laryngeal collapse associated with poll flexion in Norwegian-Swedish Coldblooded Trotter racehorses.

BACKGROUND: Dynamic laryngeal collapse (DLC) associated with poll flexion is the most common disorder of the upper respiratory tract (URT) in the Norwegian-Swedish Coldblooded Trotter (NSCT). The disorder, which has also been diagnosed in other breeds of trotters and gaited horses, appears to be related to anatomic phenotypes and only occurs during poll flexion when the horse is exercised 'on the bit'. OBJECTIVES: Identify genomic regions associated with DLC in the NSCT by combining a rigorous phenotyping protocol with genomic data from a high-density equine genotyping array. STUDY DESIGN: Prospective case/control study. METHODS: High-speed treadmill endoscopy was used to phenotype horses (n = 61) for DLC, distinguishing between cases and controls. Genome-wide association (GWA) analysis of DLC status was then performed using a principal component approach (PCA) with haplotype analyses subsequently performed for regions containing single-nucleotide polymorphisms (SNPs) above the suggestive genome-wide significance (GWS) threshold (P<1.0 × 10-5 ). RESULTS: One region containing 10 SNPs (Equus caballus chromosome [ECA] 7: 89,601,935-94,647,192) was above the suggestive GWS threshold. Two inferred haplotypes in this region demonstrated significant differences (P<0.001) between cases and controls, with the most frequent haplotype resulting in a significantly increased risk of DLC. MAIN LIMITATIONS: Small sample size as a result of stringent phenotyping protocols. CONCLUSIONS: The current study highlights a candidate genomic region on ECA7 as potentially important with regard to the manifestation of DLC. Further exploration of this region and the genes included within it will bring veterinarians and researchers closer to fully understanding the biological mechanisms underlying DLC in horses.

Competition lifespan survival analysis in the Norwegian-Swedish Coldblooded Trotter racehorse.

BACKGROUND: Studies of large racing populations have established clear differences in the career profile of stallions, mares and geldings. Multiple studies have also demonstrated positive effects on racing careers for horses that commence racing at a younger age. However, the applicability of these studies to small, native racing populations is unknown and warrants investigation. OBJECTIVES: To provide summary statistics for performance outcomes for the Norwegian-Swedish Coldblooded Trotter and to document and provide evidence on the current differences in racing careers across age at first start groups, sexes and country of birth. STUDY DESIGN: Cohort study. METHODS: Performance data on the population of Norwegian-Swedish Coldblooded Trotters born between 2000 and 2009 (n = 14,548) were acquired and used to describe and compare the racing careers of this native racing breed. Career length, career starts and career earnings were evaluated. Kaplan-Meier survival curves, stratified by sex, age group and country of birth were produced for career length. A Cox proportional hazards model was fitted to assess factors influencing the hazard rate of retirement from racing. RESULTS: Log-rank test for equality of career length survival functions showed significant differences (P<0.001) across sexes, countries of birth and age at first start groups. An increased age at first start increased the hazard rate of retirement from racing. MAIN LIMITATIONS: Racing career length is influenced by many factors. While this study accounts for some of the known influences on career length, the analysis would be strengthened by the inclusion of additional information, such as trainer and reasons for retirement. CONCLUSIONS: The results of this study suggest no adverse effects to the racing of young Norwegian-Swedish Coldblooded trotters. The study also establishes significant differences in career length based on sex and identifies that these differences are the opposite of what is seen in many other horse racing breeds. The Summary is available in Spanish - see Supporting Information.

Long-term follow-up of Norwegian horses affected with acquired equine polyneuropathy.

BACKGROUND: Acquired equine polyneuropathy (AEP), a neurological disease clinically characterised by knuckling of metatarsophalangeal joints, has been described in numerous Nordic horses during the last 20 years. Although clinical recovery has been reported, large-scale data on long-term follow-up of survivors have been lacking. OBJECTIVES: To describe long-term survival of AEP affected horses registered in Norway, with a focus on athletic performance and possible residual clinical signs connected to the disease. STUDY DESIGN: A retrospective cohort study. METHODS: The study includes 143 horses recorded with AEP in Norway from 2000 to 2012, with the follow-up period continuing until 2015. Participating owners of survivors completed a standardised questionnaire, providing information on disease and convalescence, management, performance-level and possible residual clinical signs. To investigate the follow-up of survivors, we performed 2 multivariable linear regression models. RESULTS: The follow-up time of survivors was 1.0-14.5 years (median 5.3, interquartile range 2.5-7.2). Fifty-seven horses survived and all but 3 horses returned to previous or higher level of performance. However, possible disease-related residual clinical signs were reported in 14/57 horses. Forty-nine of the survivors were in athletic use at time of contact. The majority of survivors were categorised with low severity-grades at time of diagnosis and the initial grade was significantly associated with time to resumed training. Only 3 horses had experienced relapse/new attack during the follow-up period. MAIN LIMITATION: Athletic performance was judged by owners, which renders a possible source of bias. CONCLUSIONS: Although AEP is a potential fatal disease, most survivors will recover and return to minimum previous level of athletic performance. Some horses display residual clinical signs, but often without negative effect on performance and relapse of disease is rare.

Acquired equine polyneuropathy in Norway and Sweden: a clinical and epidemiological study.

REASONS FOR PERFORMING STUDY: Acquired equine polyneuropathy (AEP, also known as 'Scandinavian knuckling syndrome'), is a serious disease of unknown aetiology, which emerged clustering in horse farms in Sweden, Norway and Finland in the 1990s. Clinical and epidemiological data regarding the syndrome are scarce. OBJECTIVES: To describe the clinical and epidemiological findings and outcome in outbreaks of AEP and compare risk factors in affected and unaffected horses on affected farms in Norway and Sweden during 2007-2009. METHODS: Neurological examinations were performed and data collected regarding demography, usage, turning-out, feeding, prophylactic strategies and long-term outcome. RESULTS: Thirteen affected farms with 157 horses of various breeds, of which 42 were AEP cases, were studied. Typical digital extensor dysfunction and knuckling of pelvic limbs were noted in 34 definitive cases. Eight additional plausible cases had a severe, acute course of neurological disease. There were no signs of brain orcranial nerve dysfunction. Cases occurred from December to April, with new cases emerging within 100 days of the index case. Affected and unaffected horses were fed wrapped forage. Prevalence for AEP was 27% and case fatality 29%. The median duration of AEP in survivors was 4.4 months (1-17 months). Survivors returned to full work within 19 months (median 6.6 months). Acquired equine polyneuropathy was less prevalent in horses aged > 12 years and young horses had a higher chance of survival than older horses. Management factors did not differ between affected and unaffected horses. CONCLUSIONS: Acquired equine polyneuropathy is a potentially fatal neurological disease characterised by pelvic limb knuckling. Surviving horses returned to normal function after a long period of rest. Cases were clustered in farms during the winter/spring season. Wrapped forage was used in all farms. POTENTIAL RELEVANCE: The results provide valuable insights into the dinical examination, handling and prognosis of cases of AEP, an emerging neurological disease of unknown aetiology in horses.

The effect of sedation on D(+)-xylose absorption tests in 6 normal horses.

REASONS FOR PERFORMING STUDY: D(+)-xylose absorption tests are commonly performed when investigating suspected small intestinal malabsorption in the horse. The test involves the administration of a D(+)-xylose solution via a nasogastric tube followed by serial blood sampling to determine its rate of absorption. In some horses, nasogastric intubation cannot be safely performed without prior administration of a sedative. Due to its short duration of action, the α(2) agonist xylazine is commonly used for this purpose. However, α(2) agonists have also been reported to influence the rate of gastric emptying as well as small intestinal motility patterns. OBJECTIVE: To evaluate if prior sedation with xylazine would influence the rate of absorption of D(+)-xylose in 6 normal Standardbred horses in a randomised cross-over study. METHODS: D(+)-xylose was administered by nasogastric intubation at a dose rate of 0.5 g/kg bwt given as a 10% solution with water while xylazine was administered iv at a dose rate of 0.5 mg/kg bwt. A heparinised blood sample was collected prior to administration of D(+)-xylose (and xylazine when used) and then at 30, 45, 60, 75, 90, 120, 150, 180 and 240 min following administration. Samples were immediately analysed using a modified colorimetric micro method. The cumulative amount of D(+)-xylose absorbed at each time point with and without prior sedation were. The significance rate was set at P<0.05. RESULTS: The study failed to demonstrate a statistically significant difference in the amount of D(+)-xylose absorbed between sedated and unsedated animals, although there was a tendency for a less rapid initial uptake with prior sedation. CONCLUSION: This study suggests that prior sedation with xylazine will not significantly affect the result of a D(+)-xylose absorption test in the normal horse.

Muscle characteristics in young Norwegian-Swedish Coldblooded Trotters and associations with breeding index, body size and early training.

REASONS FOR PERFORMING STUDY: The trotting speed of Norwegian-Swedish Coldblooded Trotters (NSCTs) is, although inferior to Standardbred speeds, gradually increasing. This development is mainly attributed to breeding progresses. Characteristics of the middle gluteal muscle have recently been described in a small number of these horses but the actual spread in muscle parameters within the population is unknown. OBJECTIVES: To describe the characteristics of the middle gluteal muscle and their naturally occurring variability in young NSCT horses and investigate possible association with body size, individual breeding index and early training. METHODS: Biopsies of the middle gluteal muscle of 144 NSCT horses born in 2005 were collected at age 17-24 months. The horses were randomly selected by stratified sampling with gender and paternal progeny as stratification factors. The muscle samples were analysed for fibre type composition, fibre type areas, capillary supply and the activity of the key metabolic enzymes citrate synthase, 3-OH-acyl-CoA-dehydrogenase and lactate dehydrogenase. Correlation analysis including muscle parameters and body size, individual breeding index and early training was performed. RESULTS: Large interindividual differences were identified in muscle characteristics. Male horses had higher percentages of type IIA fibres and lower percentages of type IIX fibres than age-matched females. Fibre type areas followed the order type IIX>IIAX>IIA>I, while the capillary supply per fibre type area increased in the opposite order. No gender-related differences were found in fibre type areas or capillary supply. Height was correlated to the relative area of type I fibres. Early training was associated with increased oxidative capacity and increased percentage of type IIA fibres. The lowest breeding index was found in untrained horses. CONCLUSIONS: Muscle characteristics differ among young NSCT horses, but the innate oxidative capacity seems to be lower than in Standardbreds of comparable age. Early training may influence on these parameters.

Effects of training on equine muscle fibres and monocarboxylate transporters in young Coldblooded Trotters.

REASONS FOR PERFORMING STUDY: Muscular changes caused by training are breed-specific and studies on the Norwegian-Swedish Coldblooded Trotter (NSCT) are limited. Knowledge about lactate-transporters in muscle in this light draught breed used for harness racing is lacking. OBJECTIVES: To identify muscular changes associated with training in young NSCTs and investigate muscular distribution of the monocarboxylate transporter 1 (MCT1) and its ancillary protein CD147, which facilitate lactate transport across membranes. METHODS: Nine horses were followed from the start of their training period until the end of their 3-year-old season. A biopsy sample of the middle gluteal muscle was collected on 4 occasions. On the last 3 sampling occasions, individual V(La4)-values (the speed corresponding to a blood lactate concentration of 4 mmol/l) were determined in an incremental exercise test on a high-speed treadmill. One horse was excluded due to lameness. Histochemical and immunohistochemical analyses were performed on all muscle samples to determine fibre types (I, IIA, IIAX, IIX), oxidative capacity (NADH) and the expression of MCT1 and CD147. The activity of selected metabolic enzymes in the muscle before and after training was determined. RESULTS: The percentage of type IIX fibres decreased with training while the percentage of type IIAX fibres increased. The activity of citrate synthase and 3-OH-acyl-CoA-dehydrogenase increased with training. The expression of MCT1 was lower in membranes and cytoplasm of type IIX fibres compared to all other fibre types both before and after training. The antibody against CD147 stained membranes and cytoplasm of all fibres. The first V(La4)-value was lower than the last 2 in all horses. CONCLUSIONS: Muscular changes with training of NSCTs were similar to those reported in Standardbreds, indicating fibre type transitions and increased oxidative capacity. Expression of MCT1 differed among fibre types and was related to the oxidative capacity of the fibres.

Polyneuropathy associated with forage sources in Norwegian horses.

BACKGROUND: Cases of hindlimb digital extensor weakness of unknown etiology have been observed in Norway since 1995. HYPOTHESIS: We hypothesized that the observed bilateral extensor weakness was attributable to neuropathy of the distal nerves and that this was related to environmental factors, possibly dietary. ANIMALS: Seventy-five horses with digital extensor weakness occurring from 1995 to 2004 are described. METHODS: Eleven horses were examined at The Norwegian School of Veterinary Science, and the medical records of 64 horses seen in ambulatory practice were reviewed. RESULTS: There was no apparent sex, age, or breed predilection, but the majority were horses kept for pleasure or breeding purposes. Clinical signs varied from intermittent knuckling of the hindlimbs to paraplegia. Some horses showed no or only slow progression of signs, whereas others developed severe signs within hours. No other neurologic deficits were detected in any of the horses. Epidemiologic data and laboratory results were not supportive of an infectious etiology. The only common factor for all affected horses seemed to be feeding big bale silage or, occasionally, hay of poor microbiologic quality. Forty of the 75 horses were euthanized. Histopathologic examination of peripheral nervous tissue was performed in 22 horses, all of which had neuronal fiber degeneration. The majority of horses with mild signs recovered after 5-6 months of rest. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinical signs correlated with polyneuropathy involving sciatic nerves.

Detection of a radiographically occult fracture of the lateral palmar process of the distal phalanx in a horse using computed tomography.

A horse with a suspected injury of the distal phalanx was examined using radiography at day two and 19 after the onset of the lameness, with no definite diagnosis. Using computed tomography an incomplete fracture of the lateral wing of the distal phalanx was diagnosed at day 25. Based on computed tomography it was determined that the fracture probably did not enter the joint or involve the palmar cortex throughout its length which were of prognostic importance. The day following the CT examination a new oblique radiographic projection was made. In this radiography which was based on, and never would have been attempted without, the information about the geometric pattern of the fracture achieved by CT examination, a radiolucent area indicative of a wing fracture was seen. This report emphasizes the value of computed tomography in diagnosing a geometric complicated nonarticular oblique fracture of the palmar process of the pedal bone in a horse.

Use of two in vitro methods for the detection of benzimidazole resistance in equine small strongyles (Cyathostoma spp.).

Ten stables were included in a study to evaluate two in vitro methods for the detection of anthelmintic resistance in cyathostomes by comparing a faecal egg count reduction test (FECRT) to a larval development assay (LDA) and an egg hatch assay (EHA). The LDA was used in seven stables and EHA in the last three. On the basis of FECR values, resistance to benzimidazoles was detected in eight of the ten small strongyle populations. Resistance to pyrantel pamoate and ivermectin was not detected. The mean concentrations that inhibited hatching in 50% of the eggs (EC50), using thiabendazole (TBZ) in an EHA, were 1.02 microM in resistant populations and 0.37 microM in susceptible or suspected resistant ones. In the LDA, TBZ concentrations preventing 50% development by first/second stage larvae to the third larval stage (LC50) were 3.8 times lower than EC50 values in resistant worm populations. Mean LC50 for morantel, levamisol, ivermectin monosaccharide and avermectin-B2 in small strongyle populations susceptible to pyrantel and ivermectin was 8.0 microM, 0.99 microM, 15.6 nM and 2.93 nM, respectively. Data on pyrantel and ivermectin resistant populations could not be obtained as no resistant populations were detected. This study concludes that in vitro tests may be useful as a supplement to FECRT for the detection of benzimidazole resistance in cyathostomes, even if reference populations to be used as controls in the assays were not available. It is suggested that EC50 values for TBZ > 0.6 microM in LDA and > 0.5 microM in EHA strongly indicate benzimidazole resistance in equine small strongyles.

A field survey on anthelmintic resistance in equine small strongyles in Norway.

A field survey at 17 stables involving 221 horses was performed to evaluate the presence of anthelmintic resistance in the equine small strongyles (cyathostomes). The horses were allocated into treatment groups, and resistance to fenbendazole (FBZ), pyrantel pamoate (PYR) and ivermectin (IVM) was tested by the faecal egg count reduction test (FECR-test). Faecal samples were collected at the time of treatment, 14 days post treatment and 90 days post treatment. Resistance to FBZ, which was defined as a faecal egg count reduction < 95%, was found in 14 out of 17 stables. In 2 of the 14 stables the egg reductions were close to the limit of 95%, 91 and 93%, respectively. In 1 stable the egg reductions indicated resistance to PYR as well as detection of resistance to FBZ, 94% reduction for PYR and 85% for FBZ. No signs of resistance were detected to IVM. The investigation was performed in late autumn and winter, and due to the climatic conditions and cleaning procedures in the stables no reinfection took place during this period. The faecal egg count reduction from treatment till day 90 post treatment was used as an expression of the effect of PYR and IVM on the early stage (hypobiotic), late third stage and fourth stage larvae in the gut wall. This was justified because there was no reinfection and because the 14 day post treatment egg counts were zero or close to zero for the PYR and IVM treatment groups. The effects of PYR and IVM on the larval stages were compared and no statistically significant differences were found.

The prevalence and epidemiology of Anoplocephala perfoliata infection in Norway.

The caecum and the adjacent 30 cm lengths of ileum and large colon of 201 horses from two different regions of Norway (Ostlandet and Trøndelag) were examined for the presence of Anoplocephala perfoliata. In all, 20% of the horses were infested with the cestode (27% in Ostlandet and 7% in Trøndelag). The mean number of worms in infected horses from the two regions was 18 and 6, respectively. Information was obtained on the age, sex, breed, type of pasture and anthelmintic treatment after the grazing season for 183 of the 20 horses. The degree of infestation with A. perfoliata was not influenced by age, breed or sex. The odds ratio (confidence limits) for using permanent pastures that had been grazed by horses for at least the 5 last years in succession was 8.8 (3.2-24.4). There was a significantly higher prevalence of A. perfoliata infestation in Ostlandet (odds ration 3.1 (1.1-9.1). The relatively low prevalence of A. perfoliata infestation in Norwegian horses compared to the reported prevalences in other countries and the differences in the prevalence between Ostlandet and Trøndelag are discussed. Gross pathological examination identified lesions such as mucosal thickening, hyperaemia and/or erosions. These lesions were found in all of the infected horses, compared with only 5% of the non-infected horses. Examination of faecal samples from 26 of the infected horses detected cestode eggs in only three cases. It was concluded that faecal examination is an inadequate method for the diagnosis of A. perfoliata infestation.

The distribution of Parascaris equorum eggs in the soil profile of bare paddocks in some Norwegian studs.

Fifteen bare paddocks, which consisted of soil only and were located on 12 different studs, were examined for their content of Parascaris equorum eggs in the upper 15 cm of the soil profile. The paddocks were classified into three different groups according to the type of soil: clayey soil (A), morainic soil (B) and gravel or gravel-like sand (C). Soil profiles were collected down to a depth of 15 cm and were divided into three layers: 0-5 cm (D1), 5-10 cm (D2) and 10-15 (D3). The eggs in each layer were counted and identified as infective or noninfective eggs. The paddocks in group C, which had good drainage conditions, had significantly lower numbers of eggs in the whole profile and in D1 and D2 than the paddocks in groups A or B. Furthermore, there was a significantly higher proportion of the total egg count present in D3 in the group C paddocks. This may have been due to a higher degree of passive transportation of eggs down in the profile in the gravel or gravel-like sand. Even though there was a significantly higher proportion of infective eggs in the soil from the group C paddocks, the lower total numbers of eggs resulted in a lower total number of infective eggs in those paddocks. The study showed that the soil type was an important factor in determining the content of P. equorum eggs in the upper layer of the soil profile in bare paddocks and consequently for the potential infestation of horses with P. equorum.