1. Introduction
The stalls of horses are bedded to absorb urine, moisture, and gases and to increase the comfort, health, and well-being of the horses. In addition, a large number of people are engaged in the horse industry as trainers, riders, stable workers, farriers, and other roles working or visiting stables for many hours daily, and being subjected to the influences of the stable environment. Most of them are young people, for example riding school pupils.
Horses in northern climatic conditions (e.g., in the Nordic countries and Canada) are exercised outdoors usually for 1 to 2 h and spend, consequently, the major part of the day (often up to 23 h) indoors. Because of this, stable air quality is of considerable importance. Furthermore, training and racing in cold weather conditions expose the vulnerable respiratory system to health problems, increasing inflammatory cells in the lungs [
1]. As a consequence, respiratory disorders are common problems, and respiratory allergy is commonly diagnosed as a condition affecting the equine lung. When the condition becomes protracted it is referred to as chronic obstructive pulmonary disease (COPD) or heaves (or RAO, recurrent airway obstruction), an animal model of asthma. Anecdotal reports suggest that the condition is rare in climates where animals are outside all year around but is common in climates where horses are stabled indoors [
2]. Clinical signs in horses with this chronic lung disease include poor athletic performance, chronic couching, purulent nasal discharge, and ultimately difficulties in breathing [
2,
3].
People working in and visiting horse stables may also be exposed to the effects of the stable air. Causes of chronic airway disease both in horses and humans usually involve exposure to excessive concentrations of airborne dust, molds, viruses, bacteria, spores, aeroallergens, and endotoxins which mostly originate from bedding and feed [
4,
5,
6,
7]. Furthermore, the inhalation of gaseous irritants such as ammonia may initiate airway obstruction and exacerbate or prolong the clinical signs of COPD in affected horses [
2] as well as humans [
3].
The effect of bedding material on the quality of stable air, both on stable dust and the ammonia concentration, is significant [
8]. The various forms of bedding in a stable and even the differences in beddings between boxes within a stable [
8] influence the stable dust and gas loads, and consequently the risk to airway health of both horses and humans.
Currently, several materials are available for the bedding of boxes in horse stables. The most common bedding materials are wood shavings, saw dust, straw, and peat. Many other materials are also used as bedding, including processed (pelleted) wooden materials and (pelleted, chopped) straw from different plants. In addition, shredded or cut paper and some plant materials (by-products), as well as woodchips are used. Each of these has individual properties, including advantages and disadvantages [
9]. Stall mats are also available, but they are usually used together with bedding because of the binding of urine. It has also been reported that horses prefer bedding material for lying down as compared with areas without it [
10].
Factors considered when selecting bedding are its availability, cost, cleanness (free from dust and foreign material) and its effect on stable air quality. The bedding material should also be easy and light to handle, to avoid excessive physical exertion in stable workers. From the point of view of horses’ health and well-being, good bedding creates a layer of insulation between the horse and cold floor, pads the hard surface, prevents bruised knees, elbows, hocks and hips, and keeps the horse clean. The bedding material also affects the behavior of horses [
11,
12,
13], for example the incidence of stereotypic behaviour. In addition, good bedding material has a better potential to be re-used e.g., in farming and horticulture [
14,
15].
The objective of this study was to examine the effects of two different bedding materials, wood shavings and peat, on the health of horses. This issue was evaluated on the basis of respiratory and overall health and quality of hooves, and by measuring stable air quality.
2. Materials and Methods
The experiment was conducted in the facilities of MTT Agrifood Research Finland (MTT, currently Natural Resources Institute Finland Luke) in the south western part of Finland (latitude 60°) under autumn/early winter (October to December) climatic conditions. The duration of the experiment was 84 days. Twelve Finnhorse brood mares (four of which were pregnant) aged between 5 and 17 years were housed in box stables in individual stalls (3 m × 3 m), divided into separate sections of the stable according to the bedding material (peat; wood shavings). The stable sections were of the same size and had an identical mechanical ventilation system. The horses were held on pasture from the beginning of June to the middle of September.
The two bedding materials were selected because they are the most common materials used for bedding in Finland. They both have a low content of harmful components when manufactured, selected and stored properly. Peat is favoured as a bedding material because of its good properties in soil improvement and good composting ability, as well as its superior capacity to bind ammonia and fluid [
13,
16,
17] compared to other materials. Both bedding materials were manufactured for use as beddings in horse stalls; peat by Vapo Ltd. (Jyväskylä, Finland) and wood shavings by Joutsenon purupaali Ltd. (Joutseno, Finland).
The horses were exercised daily in paddocks in groups for four hours, and for one hour by riding or driving during the course of the experiment. The stalls were manually cleaned by the same person between 8 and 12 am when the horses were in outdoor paddocks. All feces and wet material were removed and new bedding material was added. The depth of the bedding was about 10 cm. All removed and added bedding materials were measured by their volume.
The horses were individually fed according to their needs three times per day (morning, noon, evening) with silage/haylage (DM 26.6%–6.9%) and pelleted compounded feed (DM 88%) (Suomen Rehu Ltd., Turku, Finland) to minimize the release of airborne particles from the feeds. The diet was balanced for protein (nitrogen) intake to avoid nitrogen lost in urine and, thus, to minimize the ammonia in the stable. The forage was produced by MTT and its fermentation and hygienic quality fulfilled the criteria of good quality haylage and silage [
18]. The forage was placed on the floor.
Outdoor temperatures and weather conditions were recorded daily at 8:00 am. The average outdoor temperatures in October, November and December, respectively, were −1.7 °C (−10 to 2 °C), −3.8 °C (−14 to 3 °C) and −6.6 °C (−20 to 0 °C). According to the statistics of the Finnish Meteorological Institute the temperatures in December were quite normal, but in November the daily temperatures were highly variable, and in October the temperatures were exceptionally low.
The stable temperatures and humidity, as well as ammonia and carbon dioxide levels and amount of dust, were measured daily in both stable sections. Methane (day 0) and hydrogen sulphide (days 0 and 42) contents were measured, but because of undetectable values the measuring was not continued. Gases concentrations were measured at a height of 120 cm from the ground using an Accuro gas detection pump which draws air through sampling tubes (Dräger Safety AG, Lübeck, Germany). The measurements were carried out at 6:00 am in three boxes of each stable section; from the middle of the box at the level of the muzzle of the horse. Dust was continuously collected into dust cases that were fitted in empty boxes in both stable sections at the level of 40 cm from the ground.
Exposure of the stable workers to ammonia was evaluated with personal measurements using sampling tubes attached to the lapel of the person (Dräger Safety AG, Germany) in the middle and at the end of the experiment during the cleaning of the stalls. The measurement result was converted to correspond to an exposure period of eight hours (HTP
8h) [
19]. HTP value is the concentration that is harmful to people.
A respiratory endoscopic examination was performed three times during the study (days 0, 42 and 84), including examination of the ethmoidal region, pharyngeal openings of guttural pouches, soft palate, larynx, and trachea (symptoms = 1; no symptoms = 0). Tracheobronchial aspirates were drawn during the endoscopy and cytological and bacteriological (neutrophil cells) evaluation was carried out. The classification of the neutrophil cells in bronchoalveolar smear samples was as follows: none or some single cells (−); single cells and few small pool of cells (+); several large pools of cells (++); abundant pools of cells (+++); and an extreme abundance of cells (++++).
Blood analyses, fecal analyses and hoof quality evaluation were used as measures of health and well-being of the horses. These samples were taken with the same interval as the endoscopic examination. In addition, rectal body temperature was measured and, heart rate (with stethoscope) and respiration rate via auscultation were recorded by a veterinarian researcher. Blood samples were collected from the jugular vein, and the blood analysis consisted of hemoglobin, haematocrit, serum urea, iron, protein, and differential cell count. Bacteriology, parasites, and the pH of faeces were determined. The quality of hooves was assessed from the dry matter content of hoof horn. The hoof horn samples were collected from the hooves of front legs when the horses were in shoeing. All samples were analyzed in the clinical laboratory of MTT.
The experimental design was a randomized block design with repeated measurements. After the first endoscopy, the horses were formed into pairs based on their symptomatic similarity. The two horses of each pair were then randomly allotted to different bedding material groups (peat bedding or wood shaving bedding). The procedure was repeated until all horses were divided in the two groups. The information from the first endoscopy was excluded from the data because it was included in the animal pair-variable in the model. The data (samples from horses) were analyzed using the MIXED procedure of the SAS system with the following statistical model: Yijk = µ + pi + bj + (p × b)ij + tk + (p × t)jk + (b × t)jk + eijk, where Yijk is the observation, µ is the overall mean, pi is the random effect of ith animal pair (i = 1 … 6), bj is the fixed effect of jth bedding material (j = 1 … 2), tk is the fixed effect of the time period (k = 2 or 3), and eijk is the normally distributed error with a mean of 0 and variance δ2. Terms (p × b)ij, (p × t)jk and (b × t)jk are compound effects of factors. The best fitting covariance structure for repeated measurements was selected on the basis of the Akaike information criterion. The differences were tested with Tukey’s test. Categorical variables (neutrophil cells in tracheal mucus) and 0/1-variables were not tested statistically, but were presented descriptively, because of the small number of observations and their subjective scoring making them less informative.
In animal handling and sample collection, the European Union recommendation directives (1999/575/EU) and national animal welfare and ethical legislation set by the Ministry of Agriculture and Forestry of Finland were followed carefully. The experimental procedures were evaluated and approved by The Animal Care Committee of MTT before the study was started. The endoscopic examination was carried out and all samples from the horses were collected by a veterinarian researcher.