MOLECULAR CHARACTERIZATION OF Pasteurella multocida ISOLATE^ FROM DUCKS IN KERALA
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Date
2004
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
College of Veterinary and Animal Sciences, Mannuthy.
Abstract
Twenty-five isolates from ducks and one from fow
P. multocida using standard bacteriological procedures.
strain (LKO) obtained from IVR1 was used for comparisor
were found to be pathogenic for mice. Variation in fe
dulcitol, mannitol, sorbitol and trehalose allowed the 27
into ten biovars. Two biotypes P. multocida subsp. septlca
subsp. multocida were observed among the avian examined
uniformly sensitive to enrofloxacin, chloramphenicol
isolates from ducks and one from fowl have been sero
specific PCR assay was used to confirm the identity of
PCR on template DNA prepared from blood smears was fo|
rapid method of diagnosis for pasteurella.
Dtyped
the
could N o polymorphism within the KMT1 gene
restriction analysis of PM-PCR product with Hae III
analysis of genomic DNA of all the avian isolates w
revealed three profiles each. The fowl isolates had a
majority of the duck isolates.
prof le
were characterized as
A reference chicken
. All the avian isolates
•mentation patterns of
isolates to be grouped
, and P. multocida
All the isolates were
ind pefloxacin. Eight
as A:l. A species
isolates. Performing
und to be an extremely
be demonstrated by
Restriction endonucleases
itjh Hpa II and Hha I
that was common to
Eighty-eight per cent of the isolates carried p
profiles were observed among the isolates examined.
showed a single REP-PCR profile indicating a high level
them.
asmids. Two plasmid
All the avian isolates
of homogeneity among
avi£.n Th e outer membrane protein profiles of all the
Two protein fractions with molecular weights of 37.1f
identified as the major OMPs by SDS-PAGE.
isolates were similar.
and 26.36 kDa were
A 37.15-kDa protein was identified the major
of avian isolates as of P. multocida by Western blotting
a molecular mass of 31.33 and 26.36 kDa were also
this technique.
antige
Tw
found
gene
The OmpH gene of all the avian isolates as well
were amplified using primers designed based on
sequence of a chicken isolate of P. multocida. The am
with restriction endonucleases Dra I and Hinf I
fragments respectively. The similarity of the profiles for;
P. multocida, suggested a high degree of homogeneity
region.
Restriction analysis of the OmpH-PCR products
and B:2 with Dra I generated profiles which were distinct
three serotypes. This technique can be helpful for
serotypes of P. multocida. Studies on the OmpH gene(
cholera vaccine revealed similarity with the amplified re
isolate of P. multocida. Since this gene codes for the maj
P. multocida the vaccine can be expected to confer immu
against pasteurellosis.
serotypes A:3 and B:2
blished OmpH gene
plified product digested
•ated four and three
the avian isolates of
amongst the amplified
The sequence the OmpH-PCR product has
GenBank and has been assigned the accession No
showed 98 per cent identity with P. multocida strain X
membrane protein (OmpH) gene.
lie fraction of OMPs
o other proteins with
to be antigenic using
)f serotypes A:l, A:3
rom each other for the
erentiation of various
s) of inactivated fowl
non of the local duck
sr antigenic fraction of
-lily to ducks in Kerala
bjeen submitted to the
AY 506823. The sequence
73 (serotype A:l) outer
Description
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