Abstract
Chlamydiae are of microbiological interest because of their mode of interaction with eukaryotic host cells and their specialized life cycle with unique features of parasitism. Reports regarding prevalence of infections of Chlamydophila abortus, the causative organism for chlamydial abortions in livestock, was the basis of the study. Two isolates, one each from cattle and goat abortion along with a reference isolate, were used for characterization with Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE). Elementary bodies infected Mc Coy cells, harvested from bottle cultures were disrupted by Teflon coated magnetic pellet. Urografin-76 diluted with Tris-Potassium hydrochloride was used for purification of Elementary bodies of Chlamydophila abortus organism. On protein estimation of Elementary bodies by Biuret method, all the three isolates revealed protein concentration between 500-1000 mg/100ml, which were sufficient for electrophoresis. Ten percent of resolving gel and five percent of stacking gel of polyacrylamide in which 10g of processed isolate samples along with standard protein marker and Mc Coy cell protein (control) were electrophoresed. Using Alpha Imager Gel Documentation System, the protein bands were analyzed. Twelve bands each for local bovine isolate and reference isolate were noticed while only 10 bands were there in the caprine isolate. Additional bands of 148 kDa and 135 kDa were present in bovine isolate, compared to the reference isolate, while 152 kDa and 137 kDa bands were unique for caprine isolate.
Highlights
Chlamydial organism causes economically important diseases such as pneumonitis, polyarthritis, enteritis, encephalomyelitis, conjunctivitis and placentitis leading to abortion in domestic animals (Storz, 1971)
A - Coomassie brilliant blue stained gel after SDS-PAGE B - Representation of protein bands on each well analysed by Alpha Imager Gel Documentation System; 1- Marker, 2- Reference strain, 3- Bovine isolate, 4- caprine isolate, 5- Mc Coy cell protein
Sodium dodecyl sulphate – poly acrylamide gel electrophoresis (SDS-PAGE) was used for characterization of protein fractions of different Chlamydophila abortus isolates, following the procedures described by Laemmli (1970)
Summary
Chlamydial organism causes economically important diseases such as pneumonitis, polyarthritis, enteritis, encephalomyelitis, conjunctivitis and placentitis leading to abortion in domestic animals (Storz, 1971). In Kerala, Chlamydial organism had been isolated from pneumonic lungs in goats, aborted foetus and semen samples from bovines (Francis, 1988 and Mani et al, 2006). The organisms that cause abortion in ruminants have been characterized as Chlamydophila abortus. The present study was to compare and understand the protein profile of two local isolates of Chlamydophila abortus based on a reference isolate from Department of Veterinary Microbiology, Veterinary College, Palampur, Himachal Pradesh.
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