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Physiological Factors in the Interpretation
of Equine Hematological Profile
K. Satué1, A. Hernández1 and A. Muñoz2
1Department of Animal Medicine and Surgery, School of Veterinary Medicine,
Cardenal Herrera-CEU University, Valencia
2Department of Animal Medicine and Surgery, Equine Sport Medicine Centre,

School of Veterinary Medicine, University of Córdoba

1. Introduction
Cellular components of the blood reflect specific changes in an organ or body system or
more often, a general response of the individual to some physiological or pathological
conditions. In this chapter, we will review the main physiological factors that should be
considered when interpreting equine hematological profiles. The interpretation of the
hematological profile in conjunction with history and physical findings directs the clinician
in the selection of other diagnostic, imaging and sampling techniques. Additionally, a
hematological profile provides invaluable information concerning the severity of the disease
and the response to a treatment, and it helps in establishing a prognosis. Further, horses can
have different hematological disorders, making hematology important in equine medicine.
A special consideration should be made for equine athletes, since the assessment of a
hematological profile is pivotal in the diagnosis of a reduced performance.

2. Erythron
2.1 Introduction and specific characteristics of the equine erythrocytes
The term erythron refers to red cell precursors, the tissues in which production takes place
and mature erythrocytes themselves and its functional unit, the red blood cell (RBC). The
erythron is assessed from peripheral blood samples, by calculating the number of circulating
RBC, hemoglobin concentration (HB), packed cell volume (PCV), volumetric indices, such as
mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean
corpuscular hemoglobin concentration (MCHC), a morphological examination at
microscopic level (Messer, 1995; Kramer, 2000) and sometimes, examination of the bone
marrow (Lording, 2008). When interpreting an equine hematological profile, some specific
characteristics of this species should be kept in mind.
PCV unstable. The horse is somewhat unique compared to most other mammalians in that
the spleen is a very capacious organ, storing between 6 and 12 L of red-cell-rich blood at rest
(Persson, 1967; McKeever et al., 1993). Large numbers of RBCs temporally sequestered in the
spleen can be rapidly transferred into the circulation in response to excitement (handling,

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venipuncture, loss blood, twitching, and pain) and intense exercise (Persson, 1967; 1983).
This response is induced by the release of catecholamines and therefore, the resting PCV in
horses should be carefully assessed under different excitation levels (Persson, 1967; Schalm
and Carlson, 1982). By contrast, tranquilizers and anesthetics decreased circulating RBCs,
because of splenic sequestration (Jain, 1986). Comparable changes are not found in
splenectomized horses following excitement and strenuous exercise or tranquilization
(Kunugiyama et al., 1997).
The intensity of changes in circulating RBC in relation to spleen activity depends on
individual variations, age, breed and fitness level and in the case of exercise, duration and
intensity. The time required for RBCs to return to resting values is dependent on the degree
of the excitement, and it can vary from 40 to 60 minutes to up to several hours (Jain, 1986).
Rouleaux formation and erythrocyte sedimentation rate. Rouleaux formation is the result of
the aggregation of RBCs in linear stacks and depends on the number of RBCs and their
tendency to aggregate. Rouleaux formation is a characteristic finding in healthy horses, as a
result of weak surface changes on RBC membranes (Brockus et al., 2003). There is a positive
correlation between the rate of rouleaux formation and the rate of setting of RBCs in
anticoagulated blood (erythrocyte sedimentation rate). Rouleaux formation is accentuated in
some diseases associated with hyperproteinemia, because high concentrations of plasma
proteins, particularly fibrinogen and immunoglobulins, have an insulating effect that
reduces the RBC surface membrane charge, promoting RBC aggregation (Schalm and
Carlson, 1982; Brockus et al., 2003).
Autoagglutination can be seen in some horses without hemolysis as a result of cold
antibodies, with a maximal activity at 4-20ºC or as a result of unfractioned heparin treatment
(Monreal et al., 1995). Macroscopically, agglutination has a granular appearance and
microscopically, appears as grape-like clusters of RBCs. It should be differentiate from
rouleaux by using the saline dilution test. Typically a 1:2 dilution will disperse rouleaux but
not the autoagluttinated RBCs. Infrequently, a higher dilution (up to 1:10) may be needed to
disperse rouleaux. Agglutination causes erroneous MCV values and RBC numbers
determined by impedance, because the aggregates may interfere with the electronic or
optical evaluation of the erythrocytes. Pre-treating cell suspensions from agglutinated
heparin-treated horses with trypsin might reverse the agglutination, improving the accuracy
of cell counts (Grondin and Dewitt, 2010).
Absence of peripheral signs of regeneration. Life span of equine RBC in the circulation is
approximately 140 to 150 days (Schalm and Carlson, 1982). RBCs are released from bone
marrow as mature cells and the horse is unique in failing to release reticulocytes into
peripheral blood when there is a regenerative response to hemorrhage or hemolysis.
Therefore, morphological features indicative of regeneration, such as reticulocytes and
polychromasia are rare in equine blood films. Reticulocyte counts may be performed on
marrow aspirates in anemic horses. Values greater than 5% are consistent with accelerated
erythropoiesis. Further, in those cases where anemia derives from decreased erythropoiesis,
bone marrow examination may identify the cause and enable to carry out a definitive
diagnosis (Schalm and Carlson, 1982).
The only indication of maximally stimulated erythropoiesis in routine hematology data is
anisocytosis and increased MCV (by up to 10 to 15 fl above baseline levels for an individual

2010).1 Sampling handling In relation to the anticoagulant used. They exhibit a mild degree of anisocytosis and RBC size may differ between horse breeds. This should be done as soon as possible.. another parameter to consider is the time to conduct the blood test. Complete recovery from very severe hemolytic or hemorrhagic anemia may take approximately 1 to 2 months and 2 to 3 months respectively (Lumsden et al. with a mean diameter of 5-6 μm and a MCV of approximately 40 to 52 fl (Lassen and Swardson. Because HB is spread over a larger number of cells. 1994). resulting in altered RBC values (Rose and Hodgson. They are small. www. equine blood samples stored in EDTA at 4ºC for a maximum of 72 hrs may be adequate for blood tests (Sharif et al. 1995. The RDW is wider in healthy horses than in other species. Howell-Jolly bodies. RBC morphology.. 1982. Equine RBC is relatively small compared to other animal species. Exposure of the sample to high temperatures or direct sunlight can cause hemolysis. Kramer..2. Kramer. Increased number can be seen with enhanced erythropoiesis and with decreased or compromised splenic function (Schalm and Carlson. most hematological parameters significantly decrease. Other way to evaluate regeneration is to interpret RBC distribution width or RDW. although the regenerative capability of horses is relatively poor compared with other species (Brockus et al. assays of RBC creatine concentrations enable a more accurate evaluation of the erythropoietic response in horses. a fact that artefactually leads to increased MCV and PCV (Allen et al. in order to avoid the damage caused by storage. such as Thoroughbred racehorses or Standardbred horses have an MCV lower than other breeds. However.. Similarly.. 2. when using sodium citrate. 2003). It is a coefficient of the degree of anisocytosis of circulating RBCs. On the contrary. and normal values range between 14 and 25% (Kramer. The most common change associated with storage is an increase in the RBC size. 1988).com . Grondin and Dewitt. Grondin and Dewitt. 1975a. This parameter will be increased in anemias with significant macrocytosis and/or microcytosis. Once the blood sample has been taken. 2000. 2010). 2008).2 Physiological factors influencing erythron in horses 2. 1983. blood parameters in horses are not altered after anticoagulation in heparin-lithium or EDTA. 1986). the total surface of the red cell mass is increased. Kramer. it is not a common clinical procedure. compared with the other anticoagulants (Sharif et al. it shows a positive correlation with the reticulocyte count in bone marrow aspirates and a negative correlation with myeloid-erythroid ratio (Lording. They are nuclear remnants of DNA that occasionally are seen in healthy equine peripheral blood films. 2000). 2010). This adaptation of sport equine breeds appears to be a mean to achieve an easier gas exchange during exercise (Allen and Powell.b). 2010). round.Physiological Factors in the Interpretation of Equine Hematological Profile 575 horse) (Jain. within limitations in some hematological parameters. Mean RBC creatine concentration is significantly higher in young RBC populations (Wu et al. 2000). 2000. 1983). The response to hemolysis is greater than following hemorrhage. preferably within the first 6 hours after collection. Breeds dedicated to sport.intechopen. purple inclusions. However.

2. Kurosawa et al. 2. 1998). 1977. it has been demonstrated that there are not significant differences when comparing hematological parameters obtained using two different methods: venipuncture and intravenous catheter (May et al. new experiences during training or competition.. resulting from the actions of the sympathetic-adrenal and hypothalamic-pituitary axis (Persson. boarding at shows). It has been recommended to conduct repeated measurements. lack of sleep at shows (e.2. Excitement leads to a rise in circulating .2 Accuracy of measurements The accuracy of the determinations results from the characteristics of the equipment of analysis. HB and PCV. removal from familiar environment and social group.g. 2. allowing a more reliable interpretation of the results (Persson.5 Feeding Another factor to take into account when interpreting equine erythrogram is the food and the time of blood sampling in relation to time of feeding. In fact.3 Attitude and degree of excitement of the horse Another aspect that might influence the interpretation of RBC parameters is the attitude and the degree of excitement the horse has before and during blood withdrawal (Rose and Hodgson. 2001). stall too small for the horse to lie down and rest comfortably). 2010). but also dust and very cold or windy weather. 1993. Significant increases in PCV and total plasma proteins (TPP) are found in animals after be fed. among others (Friend. strange environments (e.576 Hematology – Science and Practice 2. This fact has been associated with loss of fluids through the saliva and other gastrointestinal fluids. 2. 1994). as it involves splenic mobilization. it is very important to know the sources of errors according to the type of equipment. presence and activity of strange horses and people at shows and increased stress levels in the handlers and rider and weaning. long-distance transport. additional care should be having in blood samples taken in maximally exercised horses in order to avoid hemolysis. changes in daily routine when traveling and at shows. The use of higher gauge needles leads to satisfactory results according to most of the authors (Jeffcott. such as exercise (whether in training or competition). 1994). 1977). There are many physiological factors that cause stress in the horse. Therefore. late night events or activity in the boarding barn. adverse environmental conditions. Recently. insufficient rest between athletic events.. This is the result of the splenic contraction produced by the release of adrenaline and noradrenaline (Kurosawa et al. Messer. 1967. 1995). Persson (1975) described a variation of up to 30% in baseline HB in three Standardbred horses in which blood samples were obtained in 7 consecutive days. Venipuncture for longer than 30 sec significantly alters the hemograma. Rose and Hodgson. confinement. The main limiting factor is the time to collect the blood sample.. as well as fluid shifts from the circulation to the gastrointestinal system (Kerr and Snow. particularly high heat and humidity. there are variations in RBC parameters in horses subjected to different nutritional regimes (Greppi et www. 1998). Similarly.intechopen. 1975. Jain. 1982).2. In our experience.4 Method of venipuncture An early study showed that the use of vacuum tubes for blood collection can cause cell damage (Archer.g.

2004). Satué et al. HB and PCV in Thoroughbreds and Arabian horses have showed decreased values in winter (Gill and Wanska. Hauss (1994) relates this variation to the influence exerted by alternation between periods of light and darkness on erythropoiesis. both in cycling and pregnant mares (Gill and Wanska. 2. Persson and Ullberg (1974) had reported that baseline hematologic values were higher in stallions than in mares and geldings. It is considered a compensatory mechanism www. After that.. Satué et al. compared to animals that live at less altitude. both in athletic and sedentary horses (Gill and Rastawicka. 1979. (2011) confirmed these results. as well as in animals which common salt is added to the food. 1979). Males have slightly higher RBC. Satué.. the results of research in this field are subjected to controversy.8 Season Season is an exogenous factor that modulates the dynamic of blood components in horses. Hernández et al. 2004). 1986). 1978. However. Gill and Rastawicka (1986) described elevations in PCV and HB overnight in comparison to light time. However. 1978.Physiological Factors in the Interpretation of Equine Hematological Profile 577 al. HB in spring was significantly higher and MCV and MCH in spring and summer were significantly lower than in other seasons. supplied 8 hrs before blood collection.. Greppi et al. while females have higher MCH and MCHC (Jain. 2.7 Gender Hematological differences linked to gender seem to have limited importance in horses. it is recommended to avoid collecting blood samples within 3 hrs of feeding a large concentrate meal or hay ration or at least ensure that samples are collected at the same time each day. autumn and winter. without modifications in MCHC (Satué et .. PCV and HB were higher in mares than in males. 2011). the patterns of seasonal changes on RBC.9 Altitude Horses subjected to high altitude have significantly higher RBC. 2. 1974)..2.2. 2008. circadian and ultradian rhythms. and they also found a significant effect of biological rhythms in TPP.2... HB and PCV. Gill and Rastawicka (1986) observed in Thoroughbred racehorses and Quarter Horses that RBC.2. The authors explained these results indicating that mares and geldings established a hypokinetic circulation with increased oxygen uptake by active muscle during exercise (Persson and Ullberg.6 Circadian biological rhythms RBC parameters exhibit diurnal infradian. However. (1996) corroborated these results. Indeed. PCV and MCV in summer were significantly higher than in spring. 2011). and dissimilar changes in ambient temperatures in different locations (Ruiz et al. 1996). 1986.. 2009). 2011). probably because of the effect of androgens on erythropoiesis. this feature was not seen during exercise in this paper. In Carthusian mares. It has been suggested that intense cold decreases RBC due to the reduction in the half-life (Ruiz et al. Gill and Kownacka. Satué et al. HB and PCV values.intechopen. Gill et al. such as the breeding season (Satué et al. Furthermore. RBC. 2004.. 2. By contrast. minor differences between adult females and males have been reported. Indeed. These variations could be related to the effect of some factors. these patterns could be subjected to a different degree of tolerance to the cold. In main lines.

1988. 1986. Most of the studies on age with hematology have focused on foals from birth to 4 years of age (Harvey et al. Hernández et al. Hernández et al. such as Standardbreds (Jain. Harvey. Cebulj-Kadunc et al. (2009) found a reduction of RBC with a compensatory increase in MCV and MCH associated with aging. (2008) and Satué et al. 2004. RBC and PCV values. MCV are high at birth and then decrease. 1988). 1988). However. These results agree with those presented for other equine breeds. even though geriatric horses have received much attention recently. These RBCs may be too small to be recognized as erythrocytes by some impedance counts. 1990). HB and PCV. which is proportionally reduced to the altitude (Wickler and Anderson. 1998). 1998. 1990). 1998). Satué. large size and high RBC. they remain in the lower limit of the adult reference internal during the first year of life (Jain. Harvey. www. 2. 1990). McFarlane et al..-3 diphosphoglycerate concentrations (Harvey. without achieving statistical significance.10 Age The influence of age on hematological parameters have been evaluated in different horse breeds (Ralston et al. 1986. (1970) found that PCV and HB were lower in foals younger than 2 years.... MCHC remains constant after birth and is similar to adult values (Harvey. McFarlane et al... Jain. McFarlane et al.. Ralston et al. Normal adult hematological values are attained at 1-2 years of age. then decline more gradually over the subsequent 2 weeks.2.. 2004) and it has been explained as the result of changes in the dynamics of maturation of the RBCs (McFarlane et al. the increase in MCV was accompanied by a slight reduction in RBC. there was a gradual increase in MCV and MCH and since HB and PCV remained unchanged during this period. 2003. Declines in these values are attributed to decreased RBC survival time. and after that. 2009). Satué et al. and enhanced delivery of oxygen to the tissues due to lower . 1988. In Carthusian pregnant mares and in Spanish Purebred horses.. Mild anisocytosis is also a typical finding in young foals (Harvey.. 1998). catecholamine release and expansion of plasma volume as adjustments of fluid balance as a result of the osmotic effect of colostral immunoglobulins. 2008. (1998) found a decreasing trend in geriatric horses. Between 3 and 4 years of age.intechopen. 1990. 2002.. 1990). decreased stimulus for erythropoietin production as a result of higher HB saturation. 1984. Lipizziano (Cebulj-Kadunc et al. They do not increase to adult values until approximately 1 year of age (Harvey. decreased iron delivery to the bone marrow. increased blood oxygen content. 1990). Grondin and Dewitt. As indicated before. This fact was linked to a reduced regenerative capacity of the bone marrow (McFarlane et al. The initial hematological changes in foals at birth are thought to be due to the increase in fetal RBC destruction. 1998. Newborn foals have RBCs of fetal origin. These parameters are reduced sharply within 12-24 hrs of life. 1993). reaching its lowest values at 3-5 months of age (Jain. 1986. probably because of the increase of the age population (McFarlane et al.578 Hematology – Science and Practice for the lower content of oxygen in the atmospheric air. needed for HB synthesis. hence generating erroneous MCV. Satué. Stewart et al. inadequate iron supplementation. increased MCV appears to be a common finding associated with aging in the horse (Ralston et al. 2000). 2010)... Microcytosis in foals has been attributed to a decrease in serum iron as a result of increased demand for growth. 2002) and wild horses (Plotka et al..

1998). Kramer. Breeds ancestrally closer have minor differences in HB. 1992). 1997). PCV as low as 24% can be found in healthy draft horses and pony breeds. www. Donkeys have similar RBC.11 Breed Breed in horses exerts a significant effect on the erythron. there should be a limit in the elevation of PCV that offsets improved oxygen-carrying capacity (Muñoz et al. depending on exercise duration and intensity (short-term high intensity or maximal exercise and long-term low intensity or submaximal prolonged exercise). 1989). several studies carried out in splenectomized horses have demonstrated a marked reduction in exercise performance (McKeever et al. 1983. This fact has been implied in the loss of performance of horses with red cell hypervolemia (Funkquist et .. 1997). 2000. Muñoz et al. Muñoz et al. because of changes in blood pressure (McKeever et al.. Additionally. HB and RBC leads to higher aerobic capacity and therefore. 1993. although a later study found a reduced RBC deformability (Geor et al. but much higher MCV (Jeffcott.intechopen. 1997).. 2010). In main lines. Light horse breeds or ‘hot-blooded breeds’ have higher RBC. If we consider the importance of increased HB in the oxygen transport capacity.. A decrease of 5-10% in plasma volume is expected in short-term and in incremental-intensity exercises (McKeever et al. RBC in blood samples obtained after this type of exercise seems to be more resistant to osmotic stress (Smith et al. This reduction is attributed to the loss of sweat in order to dissipate heat produced by muscle contraction and to the exchange of fluids between the different body compartments. 1998... 1999). HB and PCV but higher MCV.. Muñoz et al. Even though the majority of the increase in PCV in high-intensity exercises is due to the splenic contraction. 2000). there is a close relationship between increased PCV and blood viscosity. MCH and MCHC (Jain. 1999). this rise derives from the mobilization of splenic RBC under the influence of the catecholamines.. 1993. HB and PCV than ponies. Grondin and Dewitt. American miniature horses have lower RBC. 2. Thus...12 Exercise Exercise has variable effects on the erythrogram. Thoroughbreds have smaller MCV than draft horses. At the onset of the exercise.Physiological Factors in the Interpretation of Equine Hematological Profile 579 2. exercise results in increased RBC. HB and PCV. Other changes associated with brief maximal exercises are small increases in MCV and decreases in MCH and MCHC. exercise-induced fluid shifts also have a role. As a consequence.2. MCH and MCHC than other breeds (Harvey et al. fitness and training levels and environmental conditions. Indeed.. aerobic performance. 1986). it is plausible to think that increased PCV.. HB and PCV and blood volume compared to draft horses or ‘cold- blooded breeds’ (Jain. 1977). This relationship is maintained until the maximum PCV is achieved (60-65%) (Persson. Kunugiyama et al. Both the intensity and the duration of the exercise determine the magnitude of the catecholamine response (Kurosawa et al. 1998. The extent of the increase in PCV is a function of the exercise intensity in maximal exercises and in increasing-intensity exercises and a linear relationship between PCV and speed has been described (Persson. 1993. 1998). 1983).. 1984). 1993.. On the other hand. The direct effect of this increased RBC is a greater oxygen transport capacity and therefore. exercise performance (Muñoz et al.2. Further. The rise in PCV during exercise is linked to higher HB and RBC.

2010). In fact. 2. endurance-trained horses have lower resting RBC.. The increased excitability of a horse as it gets fitter could result in elevations of RBC. the increased PCV. which promotes water and electrolyte-conserving mechanisms in the kidneys and gastrointestinal tract (McKeever et al. In this case.. In Standardbred trotters. However. regular monitoring of the hemogram during training has little value for assessing the fitness of the horse. HB and RBC are good indices of dehydration. Firstly. On the other hand. 2008. stimulating RBC production. Nieto et al. because of their demeanor and nervous temperament.. One of them had laminitis. 2 had colic. Other reason that can lead to increased PCV in endurance horses is pain.13 Training Although training has limited effects on RBC parameters at rest. The advantage of plasma expansion in endurance horses is to provide extra total body water for the maintenance of cardiovascular and thermoregulatory stability during prolonged exercise in order to compensate the significant losses associated with sweating (McKeever et al... Robert et al. in our experience. is very common to find PCV as low as 30-32% in endurance healthy horses with good performance (Muñoz et al. which results in a significant reduction of racing performance. some differences are found between horses undergoing high-intensity and endurance training.2 g/dl) during a competition. 2002).. Trigo et al. 2010). We have found that endurance horses with PCV higher than 50% are disqualified from competition and some of them require intensive intravenous fluid therapy (Muñoz et al. 2010).. The increase in PCV should be equal to the increase in TPP. 2010. In fact..580 Hematology – Science and Practice On the other hand. These changes appear at the beginning of the training program. with retention of water and electrolytes after only 3 days of endurance training (McKeever et al. The second reason is the effect of a greater release of aldosterone. it is very difficult to obtain a ‘true’ basal blood sample in these horses. Decreases in PCV have been reported as a consistent finding in horses with viral respiratory tract disease and in gastric ulcers (McGowan.. 2002. 2 had heart arrhythmias. By contrast. Trigo et al. 3 were retired from competition by the owners and the remaining 5 were able to finish the competition (Trigo et al. There are two main reasons to explain these results.. associated with loss of fluids because of the quantitative importance of sweating.. www. prolonged and/or intensive training can result in an excessive increase in red cell mass. Despite these results. we found that the most rapid endurance horses in competitions can have higher PCV than the slower horses.intechopen. prolonged submaximal exercise or endurance exercise leads to moderate increase in PCV. 2009).. 2002). HB and PCV values (McKeever et al. PCV and HB than sprint- trained horses (Muñoz et al.. it is clear that hematological measurements are of little value in assessing the fitness or progress of endurance horses in training. 2010).2.. 2003). phenomenon known as red cell hypervolemia. 2010). Some authors have related the hypervolemia with overtraining (Golland et . which is considered an adaptation for a greater demand for oxygen uptake. Robert et al. Speed-trained horses have higher RBC. it has been indicated that feeding fibrous diets might increase water consumption and then plasma volume (Robert et al. In these cases where PCV increases and TPP remains unchanged. other reasons different from dehydration should be considered in order to explain these results. We studied 13 endurance horses that had increased PCV (PCV> 52%) with non-increased TPP (TPP<7. 2010.. 1993). but it is very helpful in order to detect subclinical problems that can significantly reduce exercise performance. It has been hypothesized that increased blood viscosity leads to reduced capillary perfusion and inadequate utilization of oxygen by contracting muscles. HB and PCV. 2010). 2010.

Physiological Factors in the Interpretation of Equine Hematological Profile 581 2. 2008). RBC parameters do not change (Taylor-Macallister et al. This result agrees with studies carried out in pregnant women (Bailit et al. such as phenothiazine derivatives (acepromazina. 2010). Hypotension leads to increased plasma volume. until the total blood volume is restored by releasing the attachments and fetal fluids. Zvorc et al. Satué et al. whereas a decline in RBC parameters was found in the control group. 1986).. detomidine…) significantly affect the RBC parameters. 1994. 1994. with the subsequent hemodilution and reduced RBC parameters at rest (Jain.. There is not a reasonable explanation to justify these results. These drugs lead to a relaxation of the smooth muscle in the splenic capsule. In the treated mares. 2007) and other animal species (Steinhardt et al. Satué. the release of the cells to the peripheral blood and. 2004). romifidine. Brazilian and Breton pregnant mares have described a significant increase in RBC parameters during pregnancy (Berlink et al. HB and PCV (Harvey et al. 2000. The decrease of RBC parameters in pregnancy has been associated with an absolute gain of plasma. 2004. a relative oligocythemia is found. Detlef (1985) studied the effect of iron supplementation in pregnant mares compared with untreated control mares. Near the parturition. a mild anemia appeared at the end of pregnancy (Trum.. The hypervolemia of pregnancy is necessary in order to meet the demands of the gravid uterus. 2003). 1977).. after an initial short phase of hypertension. promoting the storage of RBC (Jeffcott. RBC and HB were not changed during pregnancy. despite the increased erythropoietin concentration probably derived from placental prolactin. It has been suggested that iron deficiency anemia is common in the pregnant mare. 3. have a prolonged hypotensive effect. 2. On the other hand. lactation induces a reduction in RBC. Circulating WBC represents the outcome of the dynamic production of the bone marrow. 2008). these drugs. 2006).. The hypervolemia of pregnancy has been associated with water and sodium retention after an activation of the renin-angiotensin-aldosterone axis. although it has been hypothesized that the increased fetal metabolic requirements might condition this response (Satué. After delivery. 2005).com . the foals born from supplemented mothers had higher HB and RBC than the foals born from the untreated mares (Detlef. 1985). Carthusian.15 Administration of sedatives-tranquilizers The administration of tranquilizing compounds. Additionally... In an early study.2. 1997). chlorpromazine…) and adrenergic α-2 agonists (xylazine.14 Reproductive status The researchers that have evaluated the hematological changes resulting from pregnancy in the mare have provided controversial results. Arabian.intechopen. As the increase in RBC and HB is slower than the rise in plasma.2. Studies in Thoroughbred. Leukon The term leukon refers to the set of data derived from total and differential count of white blood cells (WBC) and the analysis of WBC morphology (Grondin and Dewitt. RBC and HB. the storage in different organs or pools. 1952). www. Further. RBC parameters increase slightly. stimulated by estrogens (Satué and Domingo.. to protect the mother and the fetus from the harmful effects of decreased venous return and to prevent the mother from suffering the adverse effects of blood loss during delivery (McMullin et al.

In the blood compartment. 1995. with clear cytoplasm. they leave the bloodstream and migrate into the tissues. Welles. so there is a continuous exchange between circulating and marginal pools (Lassen and Swardson. Equine band NEUs are less frequently seen because horses do not exhibit marked left shifts during inflammatory insults compared to dogs and cats. Prolonged storage of blood may lead to the artefactual development of hypersegmented NEUs. 1995. Marginal pool of NEU adheres to the vascular endothelium. after NEUs and the main components of the immune system. 3. myelocytes and progranulocytes) (Messer. BAS) and immature (band neutrophils. 2) Blood. metamyelocytes. both mature and immature or band NEU can be found. 2000. MON. Smith. myelocytes.intechopen. In the tissues.. monocytes. www. Welles. Band NEUs have a polymorphic nucleus. 2000). Barr bodies (sex chromatin lobe/ drumstick) can be recognized in females and resemble a small purple body attached to the nucleus by a thin chromatin strand. 2007). Welles. prepared for release into peripheral blood. metamyelocytes. NEUs are functional for 1 to 2 days. while serving as a reserve. Hyposegmented NEUs have also been reported in apparently healthy Arabian horses. 2010). After passing into the blood.2 Lymphocytes LYM are the second largest population of circulating WBCs.1 Neutrophils The release of NEU from the bone marrow into the circulated depends on the tissue demands and the production of different humoral substances. 3. 1995. They are smaller than NEUs and the other granulocytes. Welles. Further. including promyelocytes. The cytoplasm is similar to this of the mature NEU (Jain. and mature ENU. This fact facilitates the migration to the tissues. lymphocytes. Then. NEU. 2010. which were diagnosed as Pelger-Huët anomaly (Grondin et al. Idiopathic hypersegmentation of NEUs have been described in Quarter Horses that lacked evidences of clinical disease. they are fagocyted by the monocyte-macrophage system or by the mucosal surfaces (Welles. Hypersegmented NEUs are rarely seen in healthy and they have five or more lobes separated by filaments.5 times per day (Lassen and Swardson. 2000. NEU can be in the circulating pool or stored in the marginal pool (in the endothelium of several organs. There is a proliferating population of NEU. being fully mature cells (neutrophils. In cases of bacterial infection. 3) Tissues. with a less condensed chromatic pattern than the segmented NEUs. It is a unidirectional movement. the band NEUs might represent between 1 and 10% of the total WBC differential count (Welles. eosinophils. Grondin and Dewitt. without constrains.582 Hematology – Science and Practice Cells can coexist in different stages of maturation.5 hours. neutral or granules stain pink and with the nucleus polymorphic and segmented and the chromatin arranged in the form of knots. such as lungs or intestine). 10-15 μm of diameter. 2000). there is a release of chemotactic substances that promotes NEU migration from the vascular bed into the tissues. because they do not return to the peripheral circulation. LYM and basophils. Circulating NEUs have a half-life of 10. Grondin and . 2000). Three main locations of the NEU can be described: 1) Bone marrow. renewing approximately 1. 1993. In inflammatory processes. mainly in the small vessels. and then. 2000). in peripheral blood. 2000). mature NEU appear as round cells. EOS.

variable in shape nuclei (oval. with a lobulated nucleus. LYMs are made up to 38-66% T cells. with scalloped nuclear margins.e. and only a small percentage comes from central lymphoid tissues. 2010). coarse chromatin pattern and scant amount of blue cytoplasm.intechopen. T cells circulate more rapidly than B cells and migration through the splenic parenchyma is faster than through the lymph nodes (Hopkins and McConnell. a transformation that is accompanied by changes in ultrastructure. Bienzle. 2000). moderately aggregated chromatin. In circulation. 1993. 2010). reddish-orange granules in the cytoplasm. 3. they have a pale- staining Golgi zone (Latimer. because most of these cells migrate into tissues. Most of the LYMs are originated in the peripheral lymphoid tissues. located in the cell periphery and with a foamy appearance (Jain. round nucleus with a notch on some occasions (Latimer and Rackich. The amount of EOS in peripheral blood is low. 2000. a cytoplasm from blue to gray. lymphatic channels. 1999. an eccentric.3 Eosinophils EOS are cells slightly larger than neutrophils that contain large. In the tissues. 3.5 Basophils BAS are cells slightly larger than the NEUs.5 between them. the MONs mature into macrophages. broad. bilobed. The half-life of LYMs varies between 20 and 200 days (Schalm and Carlson. 2009).com . The cytoplasm can also have a few clear vacuoles of variable size. with a mean duration of transit through the blood of 30 hrs. gastrointestinal tract…The half-life of circulating EOS is about 2 to 12 hrs (Latimer. They are slightly larger than small LYMs. with a pale blue cytoplasm (Kramer. Young. i. with large amounts of granules www. although to a lesser extent than NEUs. The lobulated nucleus seldom shows fine filamentation. Welles. they distributed between the circulating and marginal pools. 2000). This ratio remains constant in different physiological states and in response to disease. such as the bronchial mucosa. Blood LYMs have the ability to recirculate in the blood. 2000). allowing amplification of the immune response (Jain. 1986. 1986).4 Monocytes MON are the largest WBC in circulation. 1984). Latimer. The half-life of macrophages ranges from several days to months (Bienzle. 1982). Grondin and Dewitt.4 hrs. larger LYMs are present. Reactive LYMs or immunocytes are rarely seen in health. in the appearance of cellular receptors or by metabolic changes. and there is not exchange at the tissue level and blood. After their production in the bone marrow. 2000. with a large. The mean circulating MON life is about 8. bone marrow and thymus. with a ratio of 1/3. Welles. 1999. and they have smooth chromatin patterns and large amounts of pale blue cytoplasm (Jain. The mature cell has a diameter of 7-12 μm. Grondin and Dewitt. horseshoe) with lacy chromatin and gray-blue cytoplasm with small azurophilic granules. lymphoid and peripheral tissues and they are able to have mitosis. Smith. 1999. Degranulated EOSs are vacuolated and are rarely seen in health (Latimer. 2000). 2000). MON are released into the bloodstream.Physiological Factors in the Interpretation of Equine Hematological Profile 583 with a dark-staining nuclei. The circulation time depends on the LYM subtype and the tissue of origin. Occasionally. 17-38% B cells with the remaining being null cells (Tizard. 1992). scant to moderate amounts of intensely basophilic cytoplasm and sometimes. 1999. often obscuring the nuclei and giving a raspberry-like appearance. 3.

There is a mobilization of the marginal pool of NEUs and/or LYM. Allen and Powell (1983) described that LYM count has higher in the evenings and lower in the mornings.584 Hematology – Science and Practice distributed irregularly.. whereas cold- blooded horses and miniature horses have ratios of 1. 2008. 1986). Stress leukocytosis is associated with cortisol release under certain stressful situations. as recently found in Spanish Purebred horses (Hernández et al.. This response appears between 2 and 4 hours after the elevation of the endogenous cortisol concentrations or after exogenous administration of corticoids (Rossdale et al. Lymphopenia is the result of LYM sequestration from lymphoid tissues and the eosinophenia derives from the marginalization of EOS in the blood vessels and the decreased release from the bone marrow (Caracostas et . In some cases. 2000). Kramer. Satué et al. 1999).. increased blood flow through the microvasculature and splenic contraction (Latimer. Harvey et al.7 and 0. It is well known that maximum cortisol concentrations appear in the morning (McKeever.6. lymphopenia and eosinopenia. the reduced ability to migrate from the blood to the peripheral tissues and the increased mobilization of the population of bone marrow reserve. On the contrary. 1986.6. This hormone induces neutrophilia without left shift. physiological leukocytosis and stress leukocytosis.3 Gender WBC and granulocytes are higher in females than in stallions.intechopen. These findings have been attributed to the circadian variations in the release of endogenous corticoids.6. These events result in leukocytosis with mature neutrophilia and/or lymphocytosis. Normal values are recovered in 24 hrs. This response has been also found after an endurance exercise and in response to a great variety of pathological processes (Welles. Neutrophilia derives from the mobilization from the marginal pool. and with an intense purple stain that vary in size and shape and can mask the nucleus (Jain. 2000).2 Time of the day In Thoroughbred racing horses. 1981..67. or high intensity exercise. 1984). respectively (Jain. 1993. These changes are transient and the marginal pool of NEUs is restored again in 20-30 min after the onset of the response and the LYM counts returned to baseline after 1 hr (Rossdale et al. 3. Welles. 3. 1983. 2011). Physiological leukocytosis refers to changes in circulating WBC associated with the intervention of the sympathetic-adrenal axis resulting from splenic contraction in cases of fear. excitement. 1982).. 2000). eosinophilia and monocytosis are also found (Snow et al. 3. 3. Welles. previous researchers performed in warm-blooded horse breeds reported higher values in males and www. 1983). 2009).0. Burguez et al.. because of a reduction in NEU adherence capacity.6 Physiological factors influencing leukogram in horses There are two main WBC responses.. 2000).. 1982.1 Breed Minor differences have been found among equine breeds in relation to WBC. with the hot- blooded horses having higher WBC compared to cold-blooded horses (Jain. Thoroughbreds and Arabian have a mean NEU/LYM ratio of 1.

6. 2010). 2008. EOSs are not routinely detected in the fetus and in foals at birth. there are no alterations in the proportions of the different WBC populations. 1990. 1995.. 2009). because the increase of NEU and decrease in LYM associated with cortisol concentrations.4 Age NEU number is low in the fetus (<1. resulting from a neutrophilia and lymphopenia (Snow et al.500/μl. 1986). Satué et al. it has been demonstrated that exhausted endurance horses had left shift in the NEUs and significantly lymphopenia (Trigo et al. 3. 2006). Allen et al.Physiological Factors in the Interpretation of Equine Hematological Profile 585 in females (Lassen and Swardson. and reach adult values at 1 year of age (Jain. 1990). 1993. These changes are likely secondary to catecholamine release and splenic contraction. Band NEU do not exceed 150/μl in healthy foals (Harvey. Hernández et al. At 3 hr after exercise. Jain. 2010).. increases after birth in response to cortisol (8. Cebulj-Kadunc et al.. 1993. Foals born at term have higher NEU count than foals born prematurely.. 1986. Cebulj-Kadunc et al.000/μl at 3 months of age.. EOSs count increases with aging due to prolonged exposure to allergens through life (Jain. Hernández et al. The NEU/LYM ratio returns to baseline values by 6 hrs after exercise (Snow et al. 2002). 1995. 2003.400/μl). MON and BAS do not seem to change with aging in horses (Harvey. 3. Horses that completed an endurance event at a faster speed have higher NEU/LYM ration than slower horses (Trigo et al. Other study failed to find significant differences between sexes (Lacerda et al..5 Exercise WBC show different responses according to the type of exercise. (2008) did not found differences in EOS count between young and adult horses. In addition.6. and this result has led to the hypothesis that EOSs may be a more sensitive indicator of training stress than other types www. LYM numbers in foals are low at birth (average 1.. The progressive trend towards lymphopenia in geriatric horses is characterized by a reducing in B cells CD4+ and CD8+. Endurance exercise is associated with leukocytosis.. Harvey. (2003) and Hernández et al. Satué. LYMs further decline during adulthood while NEU count remains the same. 2010).6.intechopen.. Additionally.000/μl) and then decrease to mean adult values (4000/μl) at about 4-6 months of age. (1998). in relation to immune senescence (Smith et al.. resulting in a higher NEU/LYM ratio in aged horses compared to foals (Jain. 1998).. 2008). Satué et al. McFarlane et al. 3. there is an increase in NEU/LYM ration. Band NEU. 1995. with a decrease in NEU/LYM ratio. Cebulj- Kadunc et al. Hernández et al. However. Some overtrained horses develop eosinophenia together with clinical signs of disease. 2008). The ratio NEU/LYM reaches values of 2/1 in geriatric horses (Jain. 2002. 1982). Sprint exercise is associated with leukocytosis because of neutrophilia but mainly because of lymphocytosis...6 Training Total WBC is unchanged during training for racing and for endurance events. Probably this is combined effect of increased circulating corticosteroids and splenic . achieving a mean of 400/μl by 4 months of age (Harvey. Lassen and Swardson. 1993. 1983). Lassen and Swardson.. 1990). increase to 5. 1990. 2004. before 300 days of gestation).

and might lead to erroneously low platelet concentrations. 2008). Equine platelets stain very lightly with Wright-Giemsa stain and sometimes can be difficult to discern on blood films. leukocytosis with neutrophilia.5-3. estrous cycle and pregnancy do not change substantially the leukogram (Berlink et al. 1997). 1999).96 106/fl respectively (Boudreaux and Ebbe.. it is more common in patients with severe gastrointestinal disease due to platelet activation by circulating endotoxins and formation of aggregates of platelets and leukocytes (Hinchliff et al. 3. Finally. The reduction in WBC in Thoroughbred mares appears during the first 4 months of pregnancy.2 Blood sample collection and analytical time Repeated venipuncture.. some studies have found a reduction in WBC. 1994). oval or elongate. measuring 2. It is advisable to perform the analysis within the first 2 hrs after collection.. 1994). EDTA-dependent pseudo thrombocytopenia has been reported in a Thoroughbred gelding (Hinchcliff et al.6 fl and 0. Platelets Platelets or thrombocytes are cytoplasmic fragments of megakariocytes. alterations in blood flow or delay in carrying out the analysis alter platelet count. It is important to take into account that decreased NEU count and later.7 Reproductive status Although in main lines.. 2000). it is www. are associated with accelerated thrombocytopoiesis. Equine platelet concentrations are some of the lowest reported for mammals.1 Physiological factors influencing platelets in horses 4.intechopen. 1993.1. 1994). this idea has not been confirmed by all the authors (Taylor-Macallister et ... The survival time of equine platelets in circulating blood is 4-7 days (Jain. 2000. Morphologically. 4.1 Anticoagulant The use of EDTA as anticoagulant. 2004.47-0. 2003).586 Hematology – Science and Practice of WBCs (Tyler-McGowan et al. lactation induces leucopenia with an intensity proportional to the degree of stress during the period of maximum milk production (Harvey et al. as MPV can be altered if the EDTA-sample is kept refrigerated. 1993). giant platelets.. greater than the diameter of a RBC. 4. Mean platelet volume (MCV) and mean platelet mass have been reported in horses: 4...3-5. 4. Platelet clumping indicates platelet activation and aggregation during blood collection.1. Saigo et al. with a trend toward increase from half of pregnancy and it is maintained until the time of delivery (Gill et al. During delivery. 1993). NEU and EOS counts during pregnancy (Satué. Satué et al. lymphopenia and eosinopenia appear. Da Costa et al. that are common causes of loss of performance in trained horses (McGowan. 1998). 1984. in association with the hypothalamic-pituitary-adrenal stimulation and glucocorticoid release (Silver et al. However. They are round. although it can produce aggregation in normal situations. 2010).6. On the other hand. Harvey et al.. Finding 6-10 platelets/field of high resolution in a peripheral blood film indicates an adequate platelet concentration.. increased LYM count is consistent with systemic or respiratory disease. 2005)..5 μm in diameter. with light blue cytoplasm with fine azurophilic granules (Kramer.

1. Satué. 2009). coupled with increased levels of thromboxane B2 produced by the placenta.5 Exercise and training The effect of exercise on platelet parameters seems to be intensity dependent. most of the studies concluded that pregnancy does not exert significant effects on circulating platelets (Harvey et al. other studies in horses did not agree with these results (McFarlane et al. as well as in other species (Zinkl et al. Hormonal dynamic during pregnancy.. 1988.4 Age Platelet numbers in foals do not change during the first year of life. chorion and amnion.. (2010) described a progressive decline in platelet numbers during pregnancy in Carthusian broodmares. 2004. 1998). 1994. although factors others than the breed should be taken into consideration. laboratory animals and female elephants. infectious and some parasitic diseases. One possible explanation for the diverse results is the modifications in blood pH and hemoconcentration. mares during delivery do not develop significant changes in platelet numbers (Harvey et al. and given many of the methodological and technical problems when working with equine platelets. By contrast. with changes in ionized calcium concentrations and platelet activity. Berlink et al. Jackson et al. possibly in association with high concentrations of estrogen. Further. 1994). 4.. age determines a progressive decrease in platelet count (Ralston et al. In addition. 1992).. This response is attributed to the combined effect of stress and increased release of estrogen. 5. 1983). 1994. In the mare. 1983). Jain. Satué. 2002). progesterone and other steroid hormones (Jackson et al.. 4. It can also provide www. 4. 1998. 2000).. 2004). However. 1992). progesterone and other steroid hormones. While in other species delivery leads to thrombocytosis (Suárez et al.Physiological Factors in the Interpretation of Equine Hematological Profile 587 interesting to use sodium citrate as an anticoagulant in order to measure platelet size (Sellon. some studies reported reduced platelet aggregability in response to high-intensity exercise (Bayly et al. Short brief or maximal exercise results in significant increases in platelet numbers. Satué (2004) and Satué et al.6 Reproductive status In human beings. lactation does not exert evident influence of circulation platelet numbers in mares (Harvey et al.intechopen.. . a marked activation of the megakaryopoiesis has been found at the end of pregnancy. it is unknown if training alters platelet function. Conclusion Hematological profile is frequently used in horses as an aid for the diagnosis and/or consequences of systemic.1. Jeffcott (1977) found that the number of platelets in this breed was higher than in other equine breeds... 4. A clear explanation for this result is lacking. In adult horses.1. Finally. whereas moderate exercise does not appear to alter platelet numbers (Bayly et al... This fact continues during the initial weeks after delivery...3 Breed In Quarter Horses. 1990). 2006). but other authors described increased aggregability and activation of platelets (Kingston et al. Satué et al.1. 1988.. 1993.

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Hematology . In common with other areas of medicine. How to reference In order to correctly reference this scholarly work. Hotel Equatorial Shanghai Slavka Krautzeka 83/A No. March.Science and Practice consists of a selection of essays which aim to inform both specialist and non-specialist readers about some of the latest advances in hematology. 2012 Hematology encompasses the physiology and pathology of blood and of the blood-forming . Hematology .intechopen. profile-of-the-horse-phisiological-factors-influencing-equine-haematology InTech Europe InTech China University Campus STeP Ri Unit 405. Hernández and A.intechopen.Science and Practice Edited by Dr. Muñoz (2012). and in many respects are driven by. a greatly improved outlook for the vast majority of patients with blood disorders and malignancies. There are now many treatment options available to the modern hematologist and. A. 596 pages Publisher InTech Published online 02. Croatia Phone: +385 (51) 770 447 Phone: +86-21-62489820 Fax: +385 (51) 686 166 Fax: +86-21-62489821 www. Hematology . Available from: http://www.). Charles Lawrie (Ed. China 51000 Rijeka. Yan An Road (West). ISBN: 978-953-51-0174- 1. happily. Dr. in both laboratory and clinic. feel free to copy and paste the following: K. Charles Lawrie ISBN 978-953-51-0174-1 Hard cover.65. InTech. advances in our scientific understanding of hematological processes under both normal and disease conditions. 2012 Published in print edition March.Science and Practice. Improvements in the clinic reflect. Satué. the pace of change in hematology has been breathtaking over recent years. Physiological Factors in the Interpretation of Equine Hematological Profile. Shanghai. Office Block.