**3. Haematology of Tiger**

#### **3.1. Erythrocytes**

230 Blood Cell – An Overview of Studies in Hematology

chemistry and urine analysis.

(Gopal, 1993).

squeeze cage.

**2. Blood collection and investigation** 

drugs. It has been experienced that the values of serum enzymes increased after 72 hrs interval of 2nd immobilization by Ketamine and Xylazine mixture (personal communication, Shrivastav 2012). Sign of anemia such as pale mucous membranes weakness, fatigue and tachycardia may be observed depending on the severity of anemia. A variety of abnormalities may be noticed by analysis of blood, bone marrow cytology, serum

Wild felids are commonly injured in territorial fight or sometimes serious injuries and internal hemorrhages occur during hunting. If blood loss is above the 50% of total volume in short period may be fetal and tiger may die due to hypovolumeic shock. Information on haematology and blood biochemistry is meagre in wild animals. However, several studies on selected haematological parameters of exotic species of captive Felids have been reported. Currier and Russell (1982) studied the higher pack cell volume in wild and captive mountain lions (*Felis concolor*) and Fowler (1986) has reviewed the haematological and biochemical profile of Felids including captive tigers whereas Jain (1986) reviewed the information of the genera *Panthera, Felis, Uncia* and *Acinonyx* concluded that blood parameters were almost similar to that of domestic cat with exception of higher concentration of plasma protein and Pack cell volume (PCV). Seal et al. (1987) have studied the haematological and biochemical profile of captive Bengal tigers with emphasis of anaesthetic effect on blood parameters. Chandranaik et.al. (2006) also studied the haematology of physically restrained tigers that were kept in squeeze cages without using anaesthetics. However, the haematological and biochemical studies were made in twelve

apparently healthy tigers in free ranges of Central India (Shrivastav et.al. 2011).

Health monitoring, assessment of health during treatment and disease diagnosis in free range tigers needs baseline data on haemato-biochemical parameters. This baseline data is important especially for comparative health assessment of felids during out breaks of diseases between sylvatic and domestic cycle vice versa. It is also required, as the tiger is on top of the sylvatic food chain and to be protected for maintaining balances in ecosystem

The collection of blood for laboratory investigations is comparatively difficult in both free range and captive tigers and only possible when animal is sedated or restrained properly in

Withstanding facts, chemical capture is comparatively safe, if accomplished by trained and experienced wildlife veterinarians. There are several drugs available for sedation. Each drug works in a different manner and is more suited to some species only. The time required for a drug to have an effect depends upon the factors such as route of administration, absorption rate, concentration and physiological status of the animal while it is difficult to generalize the choice of drug and doses (WII, 1985). It depends upon circumstances like species of the animal, age, sex, weight, location, temperature regimes in season, time of the day and The morphology of erythrocytes varies with 2 to 7.6, 7.3 ± 0.45 µm in size; appears circular, discoid, central pallor with slight anisocytosis whereas the rouleaux formation(Plate- 2) is common in tiger's blood. Chandranaik, et.al. (2006) also reported the mild anisocytosis in physically restrained tigers. However, the range and mean (with one standard deviation) of total erythrocyte count (TEC) was 4.66 to 9.15, 7.9± 1.42 million /µl. Likewise haemoglobin concentration (Hb) was obtained 9.8 to13.5, 12.8 ±1.65 mg/dl in male and 7.8 to11.5, 10.8±1.05 mg/dl in female tigers (Shrivastav et.al. 2011).

Jain (1986) defined that the rouleaux formation is associated with erythrocyte sedimentation rate (ESR) and useful for evaluation of the disease status. Shrivastav et. al. (2011) encountered ESR (14 to 26, 21 ± 4.21 /hour) and PCV (36 to 45, 38 ± 4.45 %) in free range tigers (Table 1). The consequences of ESR and PCV up and downs mostly confined to

erythrocyte osmotic fragility that increased in case of immune-mediated hemolytic anemia. Taketa et. al. (1967) have assessed the oxygen affinity of the haemoglobin is much lower in felines than that of other mammals including humans.

Tigers Blood: Haematological and Biochemical Studies 233

pathogenetic mechanism and bone marrow erythroid response ( Jain, 1986).In wild animals the clinical signs and their magnitude depends on habitat and availability of nutritive materials. Prolonged nutritional deficiencies of protein vitamins and minerals essential for erythrocytes production lead to anaemia. The type of anaemia varies with the nutritional deficiency, blood loss and the animal species involved. Despite the nutritional consequences the blood loss may be encountered through traumatic injuries, complication in blood vascular system, thrombocytopenia, and coagulation disorders. A normocytic – nonchromatic, non responsive anaemia is commonly found in association with chronic infections, chronic infectious inflammatory conditions and some type of malignancies though microcytic-

Several blood sucking parasites produce blood loss anaemia in tigers like Ancylostomes, Toxoscaris that may cause haemolytic anaemia while Trypanosomes, Babesia and Haemobartonella (*Mycoplasm haemofelis*) may alter the total blood as well as plasma volumes with acute blood loss. Chronic blood loss may lead to gastrointestinal lesions, ulcers, heavy parasitism like coccidiosis, neoplasm with bleeding into body cavity, deficiency of Vitamin

hypochromic is the sign of iron deficiencies (Jain and Kono 1975).

**Plate 1.** Tiger Blood smear stained with Modified Wright Stain x1000.

K and prothrembin etc.


**Table 1.** Haematological and Biochemical Values of Bengal tigers (*Panthera tigris tigris*)

Jain (1986) reviewed the haematological parameters of big cats including *Panthera, Felis, Uncia*  and *Acinonyx* and found that the blood composition were almost similar. Among all cats few erythrocytes had single refractile structure (Heinz body) when stained with new methylene blue stain. The Heinz body appearance in erythrocytes is the unique feature of the family Felidae (Plate -1) while they are not visible usually in blood films with Romanowsky stain (Jain,1986). The reduction in erythrocyte count (TEC) and haemoglobin concentration (Hb) are generally associated with anaemia and classified on the basis of erythrocyte morphology, pathogenetic mechanism and bone marrow erythroid response ( Jain, 1986).In wild animals the clinical signs and their magnitude depends on habitat and availability of nutritive materials. Prolonged nutritional deficiencies of protein vitamins and minerals essential for erythrocytes production lead to anaemia. The type of anaemia varies with the nutritional deficiency, blood loss and the animal species involved. Despite the nutritional consequences the blood loss may be encountered through traumatic injuries, complication in blood vascular system, thrombocytopenia, and coagulation disorders. A normocytic – nonchromatic, non responsive anaemia is commonly found in association with chronic infections, chronic infectious inflammatory conditions and some type of malignancies though microcytichypochromic is the sign of iron deficiencies (Jain and Kono 1975).

Several blood sucking parasites produce blood loss anaemia in tigers like Ancylostomes, Toxoscaris that may cause haemolytic anaemia while Trypanosomes, Babesia and Haemobartonella (*Mycoplasm haemofelis*) may alter the total blood as well as plasma volumes with acute blood loss. Chronic blood loss may lead to gastrointestinal lesions, ulcers, heavy parasitism like coccidiosis, neoplasm with bleeding into body cavity, deficiency of Vitamin K and prothrembin etc.

232 Blood Cell – An Overview of Studies in Hematology

<sup>5</sup>Erythrocyte sedimentation

<sup>7</sup>Differential leukocyte

**Blood plasma biochemistry** 

<sup>5</sup>Blood urea nitrogen

<sup>7</sup>Aspertate

<sup>6</sup>Alanine Aminotransferase

count

felines than that of other mammals including humans.

erythrocyte osmotic fragility that increased in case of immune-mediated hemolytic anemia. Taketa et. al. (1967) have assessed the oxygen affinity of the haemoglobin is much lower in

 **Haematology Unit Range Mean SE (±)**  1 Red blood corpuscles (TEC) ×106/µl 4.66–9.15 7.90 1.42 2 Total Leukocytes Count (TLC) ×103/µl 6.2–11.05 8.50 1.42 3 Haemoglobin (Hb) g/dl 7.8–13.8 12.8 1.65 4 Haematocrit (PCV) Ratio 36–45 38 2.54

rate (ESR) Hours 14–26 21 4.21 6 Icterus index (II) u/l 2– 5 2 1.51

i Neutrophils – 57–75 60 5.08 ii Lymphocytes – 18–35 30 4.56

iii Monocytes – 2– 6 0.5 1.21 iv Eosinophils – 2–6 0.4 1.30 v Basophils – 0–4 0.1 1.21

1 Albumin (ALB) g/dl 2.1–4.6 3.50 0.99 2 Total protein ( TPROT) g/dl 3.7–8.7 6.40 1.88 3 Total bilirubin TBIL) mg/dl 0.4–3.2 1.90 1.21 4 Creatinine (CRE) mg/dl 1.6–4.6 2.90 1.03

(BUN) mg/dl 6.5–48.2 27.90 13.77

(ALT) IU/L 21.2–109.0 67.88 27.84

Aminotransferase (AST) IU/L 14.4–84.0 57.96 17.27

Jain (1986) reviewed the haematological parameters of big cats including *Panthera, Felis, Uncia*  and *Acinonyx* and found that the blood composition were almost similar. Among all cats few erythrocytes had single refractile structure (Heinz body) when stained with new methylene blue stain. The Heinz body appearance in erythrocytes is the unique feature of the family Felidae (Plate -1) while they are not visible usually in blood films with Romanowsky stain (Jain,1986). The reduction in erythrocyte count (TEC) and haemoglobin concentration (Hb) are generally associated with anaemia and classified on the basis of erythrocyte morphology,

**Table 1.** Haematological and Biochemical Values of Bengal tigers (*Panthera tigris tigris*)

%

**Plate 1.** Tiger Blood smear stained with Modified Wright Stain x1000.

Tigers Blood: Haematological and Biochemical Studies 235

neutrophil numbers while eosinophil commonly not affected. Meyers-Wallen et. al. (1984) observed the young cats normally have high lymphocyte counts and hence a greater tendency to develop lymphocytosis than the adults. This observation may also be attributed in the case of tigers as they belong to the member of same family with wild habitat as an escape behavior. Increases in neutrophil numbers due to physiologic influences are more pronounced in felines than in canines because of the difference in the intravascular distribution of neutrophils. Prasse et. al.(1973) have observed 3 times mean marginal pool of neutrophils of clinically healthy cats than the circulating pool whereas in dog it was about

Neutrophils considered as first line of defense against microbial infections and are important participants in inflammatory reactions. Shrivastav, et.al. (2011) encountered 57 to 75, 60 ±5.08 % with multi-lobed nuclei of 3-5 lobes while sometimes mono-lobed nuclei with pale to slightly pink granules in the cytoplasm in free range tigers( Plate3). Chandranaik, et.al. (2006) has also reported the segmented or multi lobed nuclei while Jain (1986) studied

The changes in blood neutrophil differential count (Haden, 1935) is associated with many consequences related to infectious diseases. Several functions have been suggested for the contents of granules, as neutrophils are phagocytic cells and regulating adhesiveness and aggression hydroxyl radical formation and generation of compliment derived chemotactic factors while azurophilic granules are involved in modulation of inflammatory process (Gallin, et. al.1982). Condensation of nuclear chromatin leads to formation of darker-staining plaques separated by delicate, light-staining areas with slight brown colour cytoplasm.

Shrivastav et.al. (2011) observed eosinophils contained small, uniformly round bright eosinophilic granules almost occupying the entire and clear cytoplasm (Plate 3). These cells were encountered 2 to 6, 4 ±1.21 % in free range tigers (Table 1). The nuclei of the cells were generally less lobulated than those of the neutrophils. The eosinophils are slightly larger than neutrophils. Chandranaik, et.al. (2006) also observed the larger eosinophils larger than neutrophils and lobulated nuclei with orange cytoplasm in tigers. Jain (1986) reported the granules of the eosinophil are rod-like in domestic cats and Cheetah (*Acinonyx jubatus*) while round granules in the eosinophils of Lion and Leopard. The eosinophils are commonly seen

The basophile is a numerically insignificant but functionally important leukocyte that resemble with mast cells and it believed to share similar function as it is associated with allergic reaction, inflammatory process and immunocompitancy to the body fluids. Galli et.

the sex chromatin in few neutrophils as the drumstick lobe in the female cats.

equal or slightly greater.

**4.1. The neutrophils** 

**4.2. The eosinophils** 

**4.3. The basophiles** 

in prolonged parasitic infections or allergic disorders.

**Plate 2.** Tiger Blood smear stained with Modified Wright Stain x1000.
