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. 2016 Sep 28;41(2):538–542. doi: 10.1007/s12639-016-0846-6

Haematological and biochemical indicators of tropical theileriosis diseased cattle in wilaya of Sétif (North East Algeria)

Ouarda Ayadi 1,✉, Mohamed Gharbi 2, Mohammed Cherif Benchikh-Elfegoun 1
PMCID: PMC5447623  PMID: 28615875

Abstract

The authors investigated biochemical and haematological parameters in 40 healthy and 40 tropical theileriosis (Theileria annulata) clinically infected cattle in El Eulma region (North East Algeria). The mean haematological and biochemical parameters including the number of erythrocytes, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin concentration and mean corpuscular haemoglobin were estimated. The mean hematological and biochemical parameters including the number of leukocytes, erythrocytes, hemoglobin, hematocrit, glucose, albumin, total proteins decreased significantly (p = 0.005; 0.008; 0.03 0.048; 0.002; 0.027 and 0.018; respectively) in Montbéliard breed clinically infected with T. annulata. In all diseased animals, the concentration of total and direct bilirubin increased significantly (p < 0.001). In cross breed cattle, the erythrocytes, hemoglobin, hematocrit means decreased significantly (p = 0.027; 0.003 and <0.001, respectively). Two types of anemia were detected: a microcytic hypochromic anemia in Montbéliard breed clinically infected cattle and normocytic normochromic anemia in local and cross breed cattle. Tropical theileriosis induces severe biochemical and haematological alterations that should be taken into consideration by field veterinarians for both the diagnosis of the disease and its treatment.

Keywords: Theileria annulata, Cattle, Haematology, Biochemistry, Algeria

Introduction

Bovine tropical theileriosis (Theileria annulata infection) is a tick-borne disease transmitted by Hyalomma genus ticks. The infection is due to the presence and multiplication in leukocytes then erythrocytes of the parasite resulting in progressive and severe anemia (Gharbi et al. 2012). In the Maghreb region, this parasite is the most important summer cattle disease (Ziam and Benaouf, 2004). Tropical theileriosis induces hematologic and biochemical changes associated with anemia, these changes are related to the virulence of the strain, the infectious dose, the animal (breed, immune status), but also to climatic local factors (Mahmmod et al. 2011; Singh et al. 2001).

In Algeria, to our best of knowledge, there are no studies concerning the biochemical and hematological changes induced by T. annulata infection. Based on the presence of tropical theileriosis clinical signs, 80 cattle consisting of 40 T. annulata naturally infected and 40 clinically healthy cattle have been hematologically and biochemically studied.

Materials and methods

Study area

The present study was realized in El Eulma region, wilaya of Sétif (North East Algeria). The altitude of the region varies between 800 and 1300 m. It is a semi-arid continental region with high summer temperature with a mean temperature of 22.8 °C. Winter is rainy and cold with a mean temperature of 8.5 °C. The mean annual rainfall is 482 mm (Sétif meteorological center, 2014). The total cattle population in El Eulma region is estimated to 30,000 composed of exotic (Montbéliard, Holstein Frisian, Pie Rouge and Fleckvieh) and local breeds conducted under a semi-extensive and intensive mode for milking and beef cattle respectively (Fig. 1).

Fig. 1.

Fig. 1

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Geographical location of El Eulma region, Algeria

Animals and sampling

The present study was carried out during the tropical theileriosis transmission season (from May to September 2013). A total number of 40 healthy and 40 naturally infected cattle presenting typical clinical tropical theileriosis symptoms (fever between 40 and 42 °C, lymph node enlargement, anemia and anorexia) were included in the present survey. All the clinical cases were confirmed by positive Giemsa stained blood smears. Animals were either local, exotic (Montbéliard) or crossbred cattle, aged between 10 months and 5 years were included in the survey.

The animals were blood sampled during the morning from the jugular vein before being treated. The samples were analyzed within one hour of field collection for leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean concentration of hemoglobin (MCH), heparin, glycaemia, albumin, total proteins and total and direct bilirubin.

Laboratory assays

Collected heparinized blood samples were centrifuged and the sera were collected. Glucose, total and direct bilirubin, total protein and albumin concentrations were estimated by GLUCOSE Spinreact (BSIS17-I, Girona, Spain), BILIRUBIN Spinreact (BSDTT36, Girona, Spain), TOTAL PROTEIN Spinreact (BSIS30-E, Girona, Spain) and ALBUMIN Spinreact (BSIS02-F, Girona, Spain) kits, respectively. The ISO (International Organization for Standardization, Ed. 2004, Spain) references were used as standards. The results were read with a BA-88A Semi-Auto Chemistry Analyzer (Shenzhen, China) spectrophotometer. Hematological parameters were estimated from EDTA collected blood samples with a hematology analyzer (Pentra ES 60, Kyoto, Japan).

In order to confirm the presence of T. annulata piroplasms, Giemsa stained blood smears were prepared then examined with an optical microscope at ×1000 magnification with immersion oil.

Statistical analysis

The means were compared using Excel 11 software with the Student t test at 5 % threshold.

Results

The mean hematological and biochemical parameters including the number of leukocytes, erythrocytes, hemoglobin, hematocrit, glucose, albumin, total proteins decreased significantly (p = 0.005; 0.008; 0.03; 0.048; 0.002; 0.027 and 0.018, respectively) in Montbéliard breed clinically infected with T. annulata (Tables 1, 2) The concentration of total and direct bilirubin increased significantly in diseased animals (p < 0.001) in Montbéliard, local and crossbred infected cattle. The erythrocytes, hemoglobin, hematocrit means decreased significantly (p = 0.027; 0.003 and <0.001, respectively) in crossbred cattle.

Table 1.

Biochemical parameters in cattle with tropical theileriosis and non-diseased cattle

Breed (number) Glucose
(g/L)		 Albumin
(g/L)		 Total proteins
(g/L)		 Total bilirubin
(mg/dL)		 Direct bilirubin
(mg/dL)
Montbéliard cattle (27)
 Non diseased cattle (13) 0.79 ± 0.11 33.46 ± 5.56 79.69 ± 5.99 0.66 ± 0.22 0.04 ± 0.07
 Diseased cattle (14) 0.68 ± 0.06 29.06 ± 8.78 69.28 ± 10.06 5.52 ± 1.24 1.18 ± 0.44
 p value 0.005* 0.008* 0.03* <0.001 <0.001
Crossbreed cattle (21)
 Non diseased cattle (11) 0.75 ± 0.11 32.63 ± 3.44 78.36 ± 8.36 0.75 ± 0.20 0.05 ± 0.09
 Diseased cattle (10) 0.67 ± 0.07 30.30 ± 5.39 77.31 ± 12.38 3.89 ± 1.34 1.24 ± 0.72
 p value 0.081 0.140 0.340 <0.001 <0.001
Local cattle (32)
 Non diseased cattle (16) 0.74 ± 0.08 30.56 ± 3.30 79.81 ± 5.43 0.72 ± 0.14 0.045 ± 0.09
 Diseased cattle (16) 0.73 ± 0.18 28.92 ± 5.03 75.96 ± 21.19 2.38 ± 1.19 0.08 ± 0.42
 p value 0.420 0.156 0.242 <0.001 <0.001
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* In bolded characters: statistically significant values

Table 2.

Hematological parameters in cattle with tropical theileriosis and non-diseased cattle

Breed (number) Leukocytes
(103/mm3)		 Erythrocytes
(106/mm3)		 Hemoglobin
(g/dL)		 Hematocrit
(%)		 MCV
(µm3)		 CCMH
(%)		 MCH
(pg)
Monbéliard cattle (27)
 Non diseased cattle (13) 16.98 ± 23.2 7.05 ± 1.4 10.51 ± 1.8 31.07 ± 5.5 45.61 ± 5.9 34.37 ± 1.2 15.53 ± 2
 Diseased cattle (14) 5.42 ± 2.5 5.10 ± 1.1 7.16 ± 1.3 21 ± 4.1 39.53 ± 5.3 33.48 ± 1 14.02 ± 1.6
 p value 0.048* 0.002* <0.001 <0.001 0.002* 0.027* 0.018*
Crossbreed cattle (21)
 Non diseased cattle (11) 8.23 ± 1.96 6.97 ± 1.09 10.20 ± 0.72 30.00 ± 2.09 43,9 ± 8.52 34.23 ± 1.34 15.54 ± 2.29
 Diseased cattle (10) 10.35 ± 5.69 5.69 ± 1.04 8.8 ± 1.61 24. 89 ± 4.64 41.02 ± 3.55 35.09 ± 1.24 14.69 ± 0.99
 p value 0.172 0.027* 0.003* <0.001 0.09 0.149 0.051
Local cattle (32)
 Non diseased cattle (16) 14.46 ± 21.07 6.87 ± 1.15 9.82 ± 1.06 28.81 ± 2.66 38.93 ± 15.47 34.05 ± 0.97 15.44 ± 3.13
 Diseased cattle (16) 15.53 ± 16.25 6.53 ± 1.28 9.62 ± 1.65  27.75 ± 4.64 37.33 ± 6.26 34.48 ± 1.20 14.37 ± 1.10
 p value 0.441 0.263 0.378 0.264 0.349 0.151 0.085
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* In bolded characters: statistically significant values

Whilst, the levels of leucocytes, glycaemia, total proteins and albumin did not change significantly (p = 0.441; 0.420; 0.242 and 0.156, respectively) in local and crossbred animals (p = 0.172; 0.081; 0.340 and 0.140, respectively).

Based on the classification criteria’s of Weiss et al. (2010), the significantly decrease in MCV, MCHC, MCH levels (p = 0.002; p < 0.001; p < 0.001, respectively) showed a microcytic hypochromic anemia in Montbéliard clinically infected cattle and the non significantly changes in MCV, MCHC, MCH levels showed a normocytic normochromic anemia (p = 0.349; 0.151 and 0.085, respectively) in local and crossbred clinically infected cattle (p = 0.09; 0.149 and 0.051, respectively) (Table 2).

Discussion

According to Singh et al. (2001) and Omer et al. (2003), the decrease in the total proteins concentration was due to hypoalbuminemia and hypoglobulinaemia. These authors found a significant decrease of mean concentration of total proteins, which was similar with those estimated in Montbéliard breed cattle in present survey.

The significant decrease in the concentration of hemoglobin, hematocrit and red blood cells may be due to erythrocytes destruction by macrophages in the spleen and other mononuclear phagocyte system organs (Singh et al. 2001). In experimentally infected crossbred calves, a significant progressive decrease in the concentration of hemoglobin, hematocrit and red blood cells, leukopenia followed by leukocytosis were reported (Sandhu et al. 1998).

Several authors reported a progressive decrease in hemoglobin, hematocrit and red blood cells in acute T. annulata infections (Dhar and Gautam 1979; Sharma 1979; Mehta et al. 1988; Rayula and Hafeez 1995). A progressive decrease in hemoglobin and hematocrit associated to marked reticulocytosis were also reported by Singh et al. (2001).

In the present study, there was no significant decrease in glycaemia in infected Montbéliard breed, this result is consistent with the observations of Sandhu et al. (1998) in calves expressing tropical theileriosis. Collar and Uslu (2006) indicated that the decrease of glycaemia could be due to utilization of glucose by Theileria in the blood and hepatic dysfunction consecutive to T. annulata infection. Similar findings were reported in T. annulata-infected cattle and water buffalos (El-Deeb and Younis 2009).

In this study, the results showed a significant decrease in Montbéliard cattle leukocytes concentration compared to control animals. A leukopenia was reported by Osman and Al-Gaabary (2007) in tropical theileriosis of buffalo, similar results have been found in infected cattle (Omer et al. 2002; Bell-Sakyi et al. 2004; Aulakh and Singla 2006; Modi et al. 2015).

However, Sandhu et al. (1998) observed an immediate increase in white blood cell counts, followed by a significant decrease, several days after the beginning of the infection. Ugalmugle et al. (2010) reported 40 % cases with leucocytosis and 20 % cases with leukopenia in crossbred cattle. The insignificant leukocytosis was observed in crossbred cattle in this survey.

Leukocytosis result from lymphocytes proliferation in lymphoid organs as a defensive reaction to the invasion and multiplication of T. annulata (Modi et al. 2015) while, leukopenia may be the consequence of the destruction of white blood cells by the parasites.

In the present study, there was no difference in leucocytes, glycaemia, total proteins and albumin was found between diseased and non-infected local and crossbred cattle, this discrepancy could be explained by the higher immunization of the latters for this disease.

Indigenous breeds are more resistant against T. annulata infection than exotic breeds such as Frisian and Holstein cattle (Gharbi et al. 2014; Saleem et al. 2014). This difference in sensitivity is attributed to the difference in the immune response to produce pro-inflammatory cytokine which is higher in exotic breeds (Glass et al. 2005; Jensen et al. 2008).

The significant increase in the concentration of total and direct bilirubin could be due to two mechanisms: (1) hepatic dysfunction where a significant increase in the concentration of direct bilirubin is observed and (2) hemolytic anemia (Hooshmand-Rad 1976; Yadav and Sharma 1986; Sandhu et al. 1998; Singh et al. 2001; Omer et al. 2003).

This study revealed the presence of two types of anemia, a microcytic hypochromic observed in infected Montbéliard breeds and normocytic normochromic anemia in local and crossbred cattle. The latter results were similar to those of Sandhu et al. (1998) who reported a normocytic normochromic anemia in crossbred calves. A macrocytic hypochromic anemia was reported in Holstein (Omer et al. 2002; Durrani Kamal 2008) and crossbred cattle (Modi et al. 2015). Ganguly et al. (2015) reported a normocytic hypochromic anemia in T. annulata infected crossbred cattle. This anemia could be due to both an autoimmune reaction (Hooshmand-Rad 1976) and the effect of intra-erythrocytic piroplasms (Preston et al. 1992).

Tropical theileriosis induced important changes in hematological and biochemical parameters, which varies from one region to another and were influenced by the characteristics, the dose and virulence of the strain of T. annulata in each country. These biochemical and hematological changes should be taken into consideration by the field veterinarians in the interpretation of results and the therapeutic decision.

Acknowledgments

We thank the staff of the Laboratory of Parasitology, Ecole Nationale de M”decine Vétérinaire de Sidi Thabet, Tunisia for their help in analyzing the blood samples. The authors also thank Pr. Brarhi El Hassan, director of Institut Supérieur Vétérinaire de Constantine, University Constantine and Dr. Kars Abd El Ali for their support, and all cattle farmers who agreed to let us handle their animals.

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