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ORIGINAL PAPER
Received: 17 July 2012 / Accepted: 31 August 2012 / Published online: 12 September 2012
# Springer-Verlag 2012
Abstract Theileriosis is an economically important hemopro- lymphocytes and macrophages of the lymph nodes, spleen,
tozoal disease with high morbidity and mortality in cattle. The and skin. Molecular examination revealed that these animals
present study reported the pathological features of a natural were infected with T. annulata. The present study describes the
outbreak of tropical bovine theileriosis due to Theileria annu- clinicopathological findings of bovine tropical theileriosis in
lata in Fars Province, southern Iran. T. annulata was confirmed an unpredictable weather condition.
by the presence of T. annulata piroplasms in the blood smears
and also by polymerase chain reaction test. On necropsy, pale
mucous membranes and petechial and ecchymotic hemor- Introduction
rhages in the mucosal and serosal surfaces together with
lymphadenopathy were observed. The liver was friable, yel- Theileriosis is a tick-borne parasitic disease caused by var-
lowish, and larger than normal. Hemorrhages and punched-out ious species of Theileria (Brown 1990). Theileria annulata
ulcers were observed in the abomasal mucous membrane. is the main species responsible for the disease that causes
Severe petechial hemorrhages were seen in the skin particular- heavy economic losses and results in high mortality and
ly in the hairless areas. Pulmonary edema and emphysema morbidity, as well as production losses in many susceptible
with petechial and ecchymotic hemorrhagic foci in the lungs domestic animals specially cattle in tropical and subtropical
were evident. The main histological changes were proliferation regions (Brown 1990; Uilenberg 1995). A high prevalence
of lymphocytes in the lymph nodes and proliferation of macro- of theileriosis has been reported in the Near East and Far
phages, lymphocytes, and plasma cells in the spleen, Peyer's East, southern Europe, Africa, and Central Asia (Purnell
patches, portal tracts of the liver, and interstitial tissue of the 1978; Uilenberg 1981; Brown 1990, 1997).
kidneys. The mucous membrane of the abomasum showed The intracellular parasite T. annulata has a biphasic life
numerous multifocal areas of necrosis and ulceration, and the cycle. The first phase occurs in the arthropod vector, in most
submucosal area and lamina propria adjacent to these lesions instances Hyalomma, while the second phase occurs in the
showed hyperemia and hemorrhages, with mononuclear cell domesticated and wild ruminants (Mehlhorn and Schein
infiltration. The skin showed multifocal necrotic changes, pe- 1984). The parasite-infected ticks infect cattle through inocu-
techial and ecchymotic hemorrhages, and chronic dermatitis. lation of sporozoites during feeding (d’Oliveira et al. 1995).
The schizonts of Theileria were evident in the cytoplasm of the Concurrent with activity of the vector ticks, the disease occurs
mainly in warm months of the year (Beniwal et al. 1997).
Bovine theileriosis is endemic in various parts of Iran
A. Oryan : F. Namazi (*) : M. Razavi : R. Shahriari although its mortality is variable, and possible factors such
Department of Pathobiology, School of Veterinary Medicine,
as animal breeds (Katzer et al. 1998), annual rainfall, and
Shiraz University,
Shiraz, Iran temperature are responsible for the occurrence of enzootic
e-mail: fnamazi@shirazu.ac.ir stability or instability (Darghouth et al. 1996; Mohammad
Al-Saeed et al. 2010). The clinical signs of theileriosis
H. Sharifiyazdi
include fever, nasal and ocular discharge, weakness, anorex-
Department of Clinical Studies, School of Veterinary Medicine,
Shiraz University, ia, increased respiration, and enlargement of the lymph
Shiraz, Iran nodes; the disease leads to acute anemia and jaundice
124 Parasitol Res (2013) 112:123–127
Animals
Blood samples were collected from the ear veins in vacu- DNA extraction and PCR assay
tainers containing EDTA for the preparation of thin blood
smears. All smears were then fixed with absolute methanol, To confirm the presence of T. annulata, specific primers, T.an.
stained with Giemsa, and examined by light microscopy at F1 and T.an. R1 (forward)–TGTTTCGGTTGGTTTGTTCG
×1,000 under oil immersion for the presence of Theileria sp. TCT and (reverse)–TTCCTATTCCTTACGTACTCTGGCT,
piroplasms. The blood samples were also processed for were used to amplify 1,012 bp of the T. annulata cytochrome
DNA extraction. b gene. DNA extraction and polymerase chain reaction (PCR)
Fig. 1 Piroplasms of T. annulata in Giemsa-stained blood smear of an Fig. 3 Hemorrhages and ulcers (arrows) were detected in the mucous
infected cattle (scale bar06 μm) membrane of the abomasum in T. annulata-infected cattle
Parasitol Res (2013) 112:123–127 125
Fig. 4 Hemorrhagic areas (arrows) were observed in the skin of T. Fig. 6 Dermis of cattle infected with T. annulata. It shows chronic
annulata-infected animals dermatitis with infiltration of macrophages, lymphocytes, plasma cells,
and eosinophils (scale bar056 μm)
mucous membrane showed numerous multifocal areas of annulata showed severe lymphoid proliferation (Hervas et
erosion, necrosis, and ulceration; the submucosal area and al. 1998). It has been reported that the Theileria-infected
lamina propria adjacent to these lesions showed hyperemia cells of the hemopoietic system have been transformed and
and hemorrhages, and were heavily infiltrated with lympho- proliferated (Spooner et al. 1989), and this proliferation is
cytes, plasma cells, macrophages, and eosinophils. Fibrin related to overexpression of casein kinase II that can serve
thrombi were present in some abomasal blood vessels. The as an oncogene (Seldin and Leder 1995).
epidermis, particularly that of the hairless parts of the skin, T. annulata infection in this study was diagnosed by
showed multifocal necrotic areas, and the dermis beneath observing the intraerythrocytic forms and also Koch's blue
these necrotic foci showed chronic dermatitis with hyper- bodies in the lymphocytes of the lymph nodes, spleen, and
emia and hemorrhages together with infiltration of macro- skin. Molecular examination revealed that these animals
phages, lymphocytes, plasma cells, and eosinophils (Fig. 6). were infected with T. annulata.
The submandibular and prescapular lymph nodes revealed Although the natural occurrence of theileriosis in this re-
lymphoid proliferation together with macrophage infiltra- gion was limited to warm season, i.e., April to October, the
tion. The schizonts of Theileria were evident in the cyto- present study describes an unusual outbreak of tropical thei-
plasm of the lymphocytes and macrophages of the lymph leriosis in the winter. Tick infestation with Hyalomma was
nodes (Fig. 7), spleen, and skin. The muscle fibers of the confirmed in some, but not all infected animals. It seems that
myocardium showed mild necrosis with mononuclear cell vectors other than Hyalomma are possibly involved in the
infiltration. The lungs showed pulmonary edema and em- transmission of T. annulata from the infected cattle to the
physema together with hyperemia and hemorrhages. The uninfected ones. Based on previous studies (Gharbi et al.
blood vessels, including the cerebral vessels, were 2006), a carrier state occurs in the recovered animals; hence,
obstructed with the parasitized lymphocytes and resulted the presence of tick and also the occurrence of carrier animals
in focal infarction. The meninges were congested, and some increase the probability of natural transmission from carriers
multifocal hemorrhages were present in the cerebrum and to susceptible animals. Further studies should be performed in
cerebellum. order to elucidate the route of transmission in this unusual
Specific PCR for T. annulata amplified the expected climatic condition and to clarify the role of carriers and wild
band of 1,012 bp from the DNA extracted from blood ruminants in the epidemiology of bovine tropical theileriosis.
samples of each of the 11 heifers suspected of suffering
from theileriosis (not shown). Negative controls in the assay
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