Homology between corynebacterium psedotuberculosis isolates from goats and standard reference strain
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Date
2000
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Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy
Abstract
Corynebacterium pseudotuberculosis was isolated in pure culture on blood
agar from pus collected from lymph nodes of goats, which suffered from caseous
lymphadenitis. Three field isolates of C pseudotuberculosis were characterized
and compared with standard reference" strain based on cultural, biochemical,
toxigenicity, protein profile and restriction endonuclease digest analysis of
chromosomal DNA. The morphological and cultural characters of all the field
isolates and reference strain were similar and characteristics to the species.
Biochemical reactions employed were also similar for all the strains except
isolate F3, which fermented lactose. The biochemical characters were in confirmity
with the characters described by the earlier workers.
Three different types of toxin viz., culture filtrate (CF), whole cell lysate
(WCL) and sodium chloride extract (SCE) were tested for dermonecro toxicity in
rabbit skin. Among the preparations, the whole cell lysate produced a definite
necrosis at the point of intradermal injection followed by culture filtrate and
sodium chloride extract. The toxin preparations of the isolates invariably produced
inflammatory and necrotic changes, the degree and severity of the reactions varied
between samples.
When the three toxin preparations of the isolates and reference strain were
tested for synergistic haemolytic activity with Rhodococcus equi, the whole cell
lysate produced maximum zone of haemolysis followed by culture filtrate and
sodium chloride extract.
The proteins presented in the different preparations were analyzed by
SDS-P AGE. The protein profiles discerned by whole cell lysate, culture filtrate
and sodium chloride extract were 25,9 and 9 protein bands with ranged on masses
approximately 8-200KDa, 20-200KDa and 19-191KDa respectively. The
31.6KDa and 68 KDa proteins were found to be consistent in three toxin
preparations.
The DNA extracted from the isolates and reference strain were subjected
to restriction endonuclease digestion employing Eco RI, Barn HI, BgI II, Eco RV
and Pst I. The enzyme digest of DNA varied between the enzymes employed.
There are no observable differences between the field isolates and reference strain,
when the DNA digested with these five restriction enzymes. The enzymes Eco RI,
Barn HI and BgI I presented similar restriction pattern whereas the DNA
fragments generated by Eco RV and Pst I were different from the other three
enzymes.
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Citation
171544