Pakistan Veterinary Journal
ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE)
Accessible at: www.pvj.com.pk
Hemato-Biochemical Alterations in Cross Bred Cattle Affected with Bovine Theileriosis in
Semi Arid Zone
Imtiaz Ahmad Khan*, Ahrar Khan1, Azhar Hussain2, Asif Riaz3 and Asma Aziz2
Department of Pathobiology, PMAS-Arid Agriculture University, Rawalpindi; 1Department of Pathology, University of
Agriculture, Faisalabad; 2Veterinary Research Institute, Zarar Shaheed Road, Lahore; 3Department of Veterinary Basic
Sciences, PMAS-Arid Agriculture University, Rawalpindi, Pakistan
*Corresponding author: firstname.lastname@example.org
ARTICLE H I STOR Y
Accepted: December 26, 2010
This study was carried out to ascertain the changes in hematology and serum
constituents in cross bred cattle affected with bovine theileriosis in semi arid zone
of Pakistan. A total of 50 cross bred cows of age 2-5 years were included in the
study. Twenty animals of same age were kept as healthy controls. The affected
animals showed signs and symptoms of bovine theileriosis i.e., high rise in body
temperature, general debility, enlarged prescapular lymph nodes, mucosal
hemorrhages, conjunctivitis, etc. These animals were subjected to examination of
peripheral blood smears for the presence of Theileria annulata schizonts in infected
mononuclear cells and piroplasms in red blood cells. Significant (P≤0.05) decrease
was observed in total erythrocyte counts, packed cell volume, hemoglobin, serum
total proteins, albumin, globulins, glucose, calcium, phosphorus, cholesterol and
triglycerides concentrations in cattle affected with bovine theileriosis compared
with healthy controls, while significant (P≤0.05) increase was observed in serum
bilirubin and alanine transaminase of affected cattle compared with healthy
controls. Non significant differences were observed in serum magnesium and uric
acid concentration in both affected and healthy controls. These observations
revealed that bovine theileriosis caused by T. annulata in cross bred cattle in semi
arid zone is associated with hemato-biochemical alterations.
September 27, 2010
November 25, 2010
©2011 PVJ. All rights reserved
To Cite This Article: Khan IA, A Khan, A Hussain, A Riaz and A Aziz, 2011. Hemato-biochemical alterations in
cross bred cattle affected with bovine theileriosis in semi arid zone. Pak Vet J, 31(2): 137-140.
Pakistan is predominantly an agricultural country
with a semi-arid continental subtropical climate. It is
located between latitude 23° and 36°N and longitude 60°
and 75°E. Livestock production is being hampered due to
number of diseases and management issues including lack
of proper tick control in the country (Saddiqi et al., 2010.
Among the diseases, tick born infections are of great
importance. Bovine theileriosis
hemoprotozoan disease caused by Theileria annulata
which is transmitted by Hyalomma, tick in tropics
(Mirzaei, 2007). T. annulata is one of the most
devastating blood parasite affecting cattle, buffalo, and
sheep. The disease causes serious economic losses
through mortality and loss of productivity (Glass et al.,
2003). The disease has devastating impact on small
farmers which represent the majority of livestock owners
in endemic areas. The methods currently used to protect
is a tick-borne
against tropical theileriosis are expensive and all have
serious limitations in efficacy and sustainability
(Hasanpour et al., 2008). The prevalence, morbidity and
mortality of theileriosis are considerably high (Aktas et
Theileria enters the cattle host body during tick
feeding in sporozoitic form, which instantly enters in
leukocytes of mononuclear lineage, where they got mature
into macroschizont form. Macroschizonts develop further
into microschizonts and ultimately into merozoites, which
are liberated from the mononuclear cells. These
merozoites invade erythrocytes and develop into
piroplasms. The main symptoms shown by affected cattle
are high fever and long lasting anemia, due to intra-
erythrocytic parasitism by piroplasms (Chae et al., 1996;
Shiono et al., 2001; Nazifi et al., 2010a). The disease is
lymphoproliferative in its early phases resulting
enlargement of lymph
lymphodestructive phase which is associated with a
nodes, later on enters
Pak Vet J, 2011, 31(2): 137-140.
pronounced leukopenia. In the piroplasms phase in
erythrocytes, the parasite becomes infective for the tick.
General debility, weight loss, anorexia, high body
temperature, petechial hemorrhages on conjunctival
mucosa, enlarged lymph nodes, anemia and cough are the
predominant clinical symptoms (El-Deeb and Younis,
2009). Hematological and sero-biochemical alterations are
the indicators of severity of disease and are considered to
be good tools for the diagnosis, prognosis for effective
therapy (Col and Uslu, 2007; Nazifi et al., 2010b).
Therefore, the present study was designed and executed to
determine the changes in affected cross bred cattle with
bovine theileriosis in semi arid zone in Punjab, Pakistan.
MATERIALS AND METHODS
Experimental animals and study zone
The study was conducted (July 2008 to June 2009) in
semi arid zone in two districts (Jhelum and Mandi Baha
ud Din) of Punjab, Pakistan. A total of 50 cross bred
(Friesian X Sahiwal) cattle suffering from bovine
theileriosis were selected for study. All animals used in
this study were female ranged from 2 to 5 years of age.
These animals were reared and maintained under the
optimal field management conditions. The affected cattle
were selected on the basis of clinical signs, treatment
history and presence of piroplasms in the thin blood films
and ticks on their bodies. A total of 20 clinically healthy
(Friesian X Sahiwal) cattle of almost same age from tick-
free farms were used as a control.
All the animals were subjected to examination of
mucousal membranes (conjunctival, nasal and oral) for
hemorrhages and prescapular
enlargement. Rectal temperature of each animal suffering
from bovine theileriosis was recorded. Thin blood smears
were prepared from the ear tip of each animal showing
rise in body temperature (>102 °F), enlarged lymph
nodes, cachexia and poor general health. The smears were
air-dried, fixed with methanol, stained with 10% Giemsa
stain and examined under oil immersion objective of
microscope for piroplasms in red blood cells and infected
leukocytes for schizonts of T. annulata (Soulsby, 1982).
Collection of ticks and blood samples
Ticks were collected manually from the animals
under study. These were identified for their morphology
from various body regions in separate containers having
lid according to standard criteria (Walker et al., 2003).
About 10ml blood was directly drawn from jugular
vein in clean glass test tube. Half of the blood was shifted
to an EDTA containing test tube from each animal for
hematological studies. Hematological parameters included
were total erythrocyte count, hemoglobin concentration
and packed cell volume. Serum from each blood sample
was separated and stored at –20°C until further use. Sera
were analyzed for total proteins, albumin, globulins,
bilirubin, cholesterol, glucose, triglycerides, uric acid,
calcium, magnesium, phosphorus
transaminase concentrations using commercial kits by
colorimeteric methods using spectrophotometer.
lymph nodes for
The data thus generated were subjected to un-paired
t-test to compare the difference of hematological and
biochemical parameters at P≤0.05 using computer
software Minitab 13. Chi-square test was applied to know
the statistical difference in cured and died animals.
In the present study, 50 cross bred cattle showing
clinical signs of theileriosis were included and confirmed
by smear test for the presence of schizonts of T. annulata
in blood cells. Blood sucking ticks were collected from
different parts of the body and were identified as
Hyalomma spp. The clinical findings including high rise
in rectal temperature (103-106 °F), general debility,
enlargement of prescapular lymph nodes, hemorrhages on
mucosal membranes (conjunctival, nasal, and oral),
cachexia, dyspnoea, lacrimation, conjunctivitis and eye
ball protrusion were recorded. These animals were treated
with Buparvaquone (ButalexTM 50mg/ml injection ICI,
Pakistan Limited) @ 1ml for 20 kg BW and was repeated
after 48 hours. The ratio of 44 (88.0%) cured animals was
significantly higher than those (12.0%) died (χ²= 20.574;
df =1, P-Value = 0.001).
Significantly (P≤0.05) decreased total erythrocyte
counts, packed cell volume
concentration were observed in infected cattle as
compared to non infected controls (Table 1). In infected
cattle significantly (P≤0.05) decreased serum total
proteins, albumins, globulins,
phosphorus, cholesterol and triglycerides concentrations
compared with negative controls were recorded.
Significant (P≤0.05) increase was observed in serum
bilirubin and alanine transaminase concentrations in
infected cattle compared with non infected control group.
Non-significant difference was observed in serum
magnesium and uric acid concentrations compared with
non infected control group (Table 1).
Table 1: Different hematological and biochemical parameters
(Mean±SD) in cross bred cattle affected with bovine theileriosis
in semi arid zone
Total Erythrocyte Counts 1012/L
Packed Cell Volume
Serum biochemical parameters
Values with asterisk in a row differ significantly (P≤0.05) from
that on non-infected group.
U/L 101.16±11.21* 65.52±12.24
mg/dl 101.08±17.43* 146.05±23.61
mg/dl 11.035±3.85* 17.605±4.14
Pak Vet J, 2011, 31(2): 137-140.
Anaplasma are considered to be the most important blood
parasites of cattle and buffaloes in Pakistan (Khan et al.,
2004). Bovine theileriosis of cattle is a major constraint in
the improvement of the livestock industry in larger parts
of the World (El-Deeb and Younis, 2009). Theileriosis
caused by T. annulata is a serious hemoprotozoan disease
of cattle in tropics and sub-tropics. T. annulata is
transmitted by ticks of the genus Hyalomma (Mirzaei,
Blood smears revealed
morphology along with presence of theileria schizoints in
infected mononuclear cells and erythrocytes in the present
study. The infected erythrocytes appeared as echinocytes
thorn-like protrusions on the surface. These variations in
erythrocytic morphology are mainly due to theileria
schizoints, erythrocytic oxidation, intravascular thrombi,
and immune-mediated processes (Stockham et al., 2000).
In the present study, total erythrocyte counts, packed cell
volume percentage and hemoglobin concentrations were
significantly lower in infected animals as compared with
non-infected cattle. Such findings have already been
reported by Col and Uslu (2006) and Hasanpour et al.
(2008). The decrease in erythrocyte count, packed cell
volume and hemoglobin concentration resulted anemia in
infected group. It has been reported that anemia occurs in
lateral stages of theileriosis following parasitaemia
(Mbassa et al., 1994). Immune-mediated mechanism like
erythrophagocytosis might be responsible for the
destruction of erythrocytes infected with theileria
schizoints (Uilenberg, 1981). Removal of piroplasm
infected erythrocytes by
reticuloendothelial system has been suggested as a cause
of anemia (Campbell and Spooner, 1999). The decreased
erythrocyte counts could be attributed to increased levels
of activated complement products (Omer et al., 2002).
Moreover, oxidized erythrocytes are destroyed by
erythropagocytosis, oxygen free radicals may also be
responsible for anemia (Mbassa et al., 1994).
In the present study serum total protein, albumin and
globulin concentrations in infected cattle were decreased.
The findings were in line with the findings of Col and
Uslu (2007) who reported hypoproteinemia and
hypoalbuminemia. The results were in agreement with the
findings of Yadav and Sharma (1986) and Singh et al.
(2001). The decreased serum proteins concentrations may
be attributed to extra-vascular proteinaceous fluid in body
cavities due to diseased lymph nodes resulting edema
(Stockham et al., 2000). The decrease in total serum
proteins was attributed to low albumin and globulin
concentrations as a result of liver failure (Omer et al.,
2003). In the present study serum bilirubin concentration
was increased in cattle infected with T. annulata
compared with healthy controls. The findings were in
accordance with the findings of Col and Uslu (2007) who
also reported an increased serum bilirubin concentration.
The increased serum bilirubin concentration may be
attributed to hemolysis of parasitized erythrocytes in
reticuloendothelial system and lymph nodes (Sandhu et
al., 1998). Further more, it has been reported that the
increase in serum bilirubin concentration may be due to
Theileria, Babesia and
macrophages in the
hemolytic anemia and hepatic dysfunction (Omer et al.,
2003). Serum AST and ALT concentrations are the
indicators of hepatic function (Forsyth et al., 1999). In
present study an increase in ALT concentration was
observed in infected cattle compared with healthy controls
which represent hepatic dysfunction. Similar findings
were observed by Col and Uslu (2007) who also reported
an increase in ALT concentration. The rise in serum ALT
concentration may be due to muscular trauma as a result
of prolonged recumbency due to bovine theileriosis (Col
and Uslu, 2007).
In the present study, there was a decrease in serum
glucose concentrations in infected cattle with T. annulata
compared with healthy controls. The findings were in line
with the findings of Col and Uslu (2007), they reported
that, the decreased glucose serum concentration could be
due to utilization of glucose by theileria in the blood and
hepatic dysfunction as a result of theileria infection. The
findings were contrary with the findings of Sandhu et al.
(1998) who reported a non significant decrease in serum
glucose in calves affected with theileriosis. There were
decreased serum calcium and phosphorus concentrations
in cattle infected with T. annulata in the present study
which was also decreased in the previous studies (Omer et
al., 2003). Col and Uslu (2007) reported that
hypocalcaemia in their study could be due to
hypoproteinemia, decreased dietary intake, intestinal
malfunction and kidney damage and the decreased
phosphorus concentration due to diarrhea and renal
wasting. Non significant differences were observed in
serum uric acid and magnesium concentrations in infected
and healthy controls, while previous studies revealed
increased urea level in infected cattle with T. annulata
(Sandhu et al., 1998). There were significant decreases in
serum cholesterol and triglycerides concentrations in
theileria infected cattle as in previous findings reported by
Singh et al. (2001). These decreases in cholesterol and
triglyceride levels may be ascribed to anorexia and
diarrhea (Col and Uslu, 2007). The findings were
otherwise than the findings of Yadav and Sharma (1986)
who recorded a marked increase in serum cholesterol
concentration in experimentally T. annulata infected cattle
from day 0 to day 15, followed by a sudden fall, reaching
values below the pre-infection level by day 40. They
reported that, this could be due to liver damage that results
in a concurrent increase in the level of fats with the
reduction of sugar and protein.
One the basis of present study’s findings, it was
concluded that bovine theileriosis caused by T. annulata
in cross bred cattle in semi arid zone greatly influenced
the hemato-biochemical constituents. These changes may
provide help to understand the disease pathogenesis and
could be used as a tool for diagnosis, prognosis and
evaluation of the treatment given.
Aktas M, N Dumanli and M Angin, 2004. Cattle
infestation by Hyalomma ticks and prevalence of
Theileria in Hyalomma species in the east of Turkey.
Vet Parasitol, 119: 1-8.
Campbell JDM and RL Spooner, 1999. Macrophages
behaving badly: infected cells and subversion of
Pak Vet J, 2011, 31(2): 137-140. Download full-text
immune responses to Theileria annulata. Parasitol
Today, 15: 10-16.
Chae JS, JM Lee, OD Kwon, JH Park and KS Chae, 1996.
Rapid detection of Theileria sergenti by the
polymerase chain reaction in Korean cattle. Korean J
Vet Res, 36: 195-207.
Col R and U Uslu, 2006. Haematological and coagulation
profiles during severe tropical theileriosis in cattle.
Turk J Vet Anim Sci, 30: 577-582
Col R and U Uslu, 2007. Changes in selected serum
components in cattle naturally infected with Theileria
annulata. Bull Vet Inst Pulawy, 51: 15-18.
El-Deeb WM and EE Younis, 2009. Clinical and
biochemical studies on Theileria annulata in
Egyptian buffaloes (Bubalus bubalis) with particular
orientation to oxidative
relationship. Vet Parasitol, 164: 301-305.
Forsyth LMG, FC Minns, E Kirvar, RE Adamson, FR
Hall, S McOrist, CGD Brown and PM Preston, 1999.
Tissue damage in cattle infected with Theileria
cytokineproducing, schizont-infected mononuclear
phagocytes. J Comp Path, 120: 39–57.
Glass EJ, SC Craigmile, A Springbett, PM Preston, E
Kirvar, GM Wilkie, PD Eckersall, RF Hall and CGD
Brown, 2003. The protozoan parasite, Theileria
annulata, induces a distinct acute phase protein
response in cattle that is associated with pathology.
Int J Parasitol, 33: 1409-1418.
Hasanpour A, GA Moghaddam and A Nematollahi, 2008.
Biochemical, hematological and Electrocardiographic
changes in buffaloes naturally infected with Theileria
annulata. Korean J Parasitol, 46: 223-227.
Khan MQ, A Zahoor, M Jahangir, and MA Mirza, 2004.
Prevalence of blood parasites in cattle and buffaloes.
Pakistan Vet J, 24: 193-195.
Mbassa GK, O Balemba, RM Maselle and NV Mwaga,
1994. Severe anaemia due to haematopoietic
precursor cell destruction in field cases of East Coast
Fever in Tanzania. Vet Parasitol, 52: 243-256.
Mirzaei M, 2007. Treatment of natural tropical theileriosis
with the extract of the plant Peganum harmala.
Korean J Parasitol, 45: 267-271.
Nazifi S, SM Razavi, M Reiszadeh, Z Esmailnezhad, M
Ansari-Lari and F Hasanshahi, 2010a. Evaluation of
the resistance of indigenous Iranian cattle to Theileria
annulata compared with
measurement of acute phase proteins. Comp Clin
Pathol, 19: 155-161.
stress and ketosis
by metastasis of
Holstein cattle by
Nazifi S, SM Razavi, M Reiszadeh, Z Esmailnezhad and
M Ansari-Lari, 2010b. Diagnostic values of acute
phase proteins in Iranian indigenous cattle infected
with Theileria annulata. Vet arhiv, 80: 205-214.
Omer OH, KH El-Malik, M Magzoub, OM Mahmoud,
EM Haroun, A Hawas and HM Omar, 2003.
Biochemical profiles in Friesian cattle naturally
infected with Theileria annulata in Saudi Arabia. Vet
Res Commun, 27: 15-25.
Omer OH, KH El-Malik, OM Mahmoud, EM Haroun, A
Hawas, D Sweeney and M Magzoub, 2002.
Haematological profiles in pure bred cattle naturally
infected with Theileria annulata in Saudi Arabia. Vet
Parasitol, 107: 161-168.
Sandhu GS, AS Grewal, A Singh, JK Kondal, J Singh and
RS Brar, 1998. Haematological and biochemical
studies on experimental Theileria annulata infection
in crossbred calves. Vet Res Commun, 22: 347-354.
Shiono H, Y Yagi and P Thongnoon, 2001. Acquired
methemoglobinemia in anemic cattle infected with
Theileria sergenti. Vet Parasitol, 102: 45-51.
Saddiqi HA, Z Iqbal, MN Khan and G Muhammad, 2010.
Haemonchus contortus in a natural pasture infection.
Int J Agric Biol, 12: 739–743.
Singh A, J Singh, AS Grewal and RS Brar, 2001. Studies
on some blood parameters of crossbred calves with
experimental Theileria annulata infection. Vet Res
Commun, 25: 289-300.
Soulsby EJL, 1982. Helminthes, Arthropod and Protozoa
of Domesticated Animals. 7th Ed, Baillere Tindall.
Stockham SL, AM Kjemtrup, PA Conrad, DA Schmidt,
MA Scott, TW Robinson, JW Tyler, GC Johnson, CA
Carson and P Cuddihee, 2000. Theileriosis in a
Missouri beef herd caused by Theileria buffeli: case
report, herd investigation, ultrastructure, phylogenetic
analysis, and experimental transmission. Vet Pathol,
Uilenberg G, 1981. Theileria infections other than east
coast fever. Curr Top Vet Med Anim Sci, 6: 411-427.
Walker AR, A Bouattour, JL Camicas, A Estrada-Pena,
IG Horak, AA Latif, RG Pegram, and PM Preston,
2003. Ticks of domestic animals in Africa: a guide to
identification of species.
Yadav CK and NN Sharma, 1986. Changes in blood
chemical components during experimentally induced
Theileria annulata infections in cattle. Vet Parasitol,
of sheep breeds to