Global Veterinaria 12 (6): 865-868, 2014
ISSN 1992-6197
© IDOSI Publications, 2014
DOI: 10.5829/idosi.gv.2014.12.06.8477
Corresponding Author: Agus Setiyono, Pathology Division, Department of Clinic, Reproduction and Pathology,
Faculty of Veterinary Medicine, Bogor Agricultural University,
Jln. Agatis, Kampus IPB Darmaga, Bogor 16680, Indonesia.
865
Immunohistochemical Detection of Coxiella burnetii
in Ruminants: A Case Study of Q Fever in Indonesia
Agus Setiyono and Mawar Subangkit
Pathology Division, Department of Clinic, Reproduction and Pathology, Faculty of Veterinary Medicine,
Bogor Agricultural University, Jln. Agatis, Kampus IPB Darmaga, Bogor 16680, Indonesia
Abstract: Q fever is a zoonotic disease caused by Coxiella burnetii, an obligate intracellular bacteria. Diagnosis
of Q fever is usually determined by serological test and it should be time consuming. Detection of C. burnetii
antigen immunohistochemically appears to be one of the best choice method for establishment of Q fever
diagnosis in animals. The aim of this study was to detect the present of aetiological agent of Q fever using
peroxide base method in Idul ‘Ied celebration cattles in Jakarta region, Indonesia. A total of 40 samples were
collected during 2012 and examined. Gross lesion of the liver samples revealed multifocus necrotic hepatitis,
granulomatous hepatitis, cirrhosis and fatty liver degeneration of parenchymal cells. The result showed positive
immunoreactive to C. burnetii and occurs in 3 out of 40 liver samples.This is the first report on C. burnetii
infection case diagnosed immunohistochemicallyin ruminants in Indonesia.
Key words: Antibody Coxiella burnetii Diagnosis Immunohistochemistry Ruminants
INTRODUCTION MATERIALS AND METHODS
Query (Q) fever is a worldwide zoonosis [1] caused
by Coxiella burnetii, an obligate intracellular and highly
infectious bacterium that can lives in the monocyte or
machrophage [2]. The most common syndromes observed
in acute Q fever are prolonged fever of unexplained origin, emulsified immunogen incomplete freund’s adjuvant
granulomatous hepatitis and atypical pneumonia [3, 4].
Liver involvement is common in acute Q fever and
percutaneous liver biopsy can help in the diagnosis. On
the other hand,cases of chronic Q fever hepatitis are
rarely reported in the literature [5] and are frequently
associated with endocarditis [6-8]. Diagnosis of Q fever
can be established serologically by using fluorescent
antibody technique [9-11] and or by polymerase chain
reaction to detect genetic material deoxyribo nucleic acid
of C.burnetii [12]. However, those techniques need more
time and technically laborious.Immunohistochemistry
(IHC) method had developed and thought to be a
diagnostic tool for detecting antigen in the organ or
tissue. The aim of this study is diagnose Q fever
immunohistochemically in cattle which use for Idul ‘Ied
celebration in Jakarta region, Indonesia.
Production of anti-Coxiella burnetii Antibody:
Production of antibody used immunogen formalin-fixed of
C.burnetii-Nine Mile Strain according to the standard
procedure [13] with minor modification. Briefly, the
(CFA) were applied subcutaneously to New Zealand
rabbit and followed with booster intramuscularly 3 weeks
later using emulsified immunogen-incomplete freund’s
adjuvant(IFA). Two to three weeks after boosting, the
level of anti-C.burnetii antibody was checked and the
sera considered to be harvested if the antibody titre
significantly elevated. The generated antibody was then
characterized and kept into some aliquots and store in-
20°C until use.
Immunohistochemistry: 1 x 10 purified C.burnetii cells
6
were applied intraperitoneally to Hamster. After three
weeks, Hamster was sacrificed and the spleen and liver
were harvested. To evaluate the immunoreactivity of
produced anti-C.burnetii antibodies toward certain
C.burnetii antigen in the spleen or liver of Hamster,