SERO EPIDEMIOLOGY AND MOLECULAR DIAGNOSIS OF BRUCELLOSIS IN CATTLE AND MAN IN TELANGANA AND ANDHRA PRADESH
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Date
2015-10-26
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PVNR TVU
Abstract
ABSTRACT
Epidemiology of brucellosis in Telangana and Andhra Pradesh States was
studied using cultural, serological (RBPT, LFA, STAT and ELISA), PCR assay
(Conventional, multiplex and Real time PCR) and milk tests (MRT, LFA and m-
ELISA).
Out of 2668 cattle serum samples screened from three regions, 5.81%, 5.92%,
5.10% and 6.22% were positive by RBPT, LFA, STAT and ELISA respectively. Out of
97 males, 5.15%, 5.15%, 3.85% and 5.15% and out of 2571 females, 5.83%, 5.95%,
5.21% and 6.26% were positive by RBPT, LFA, STAT and ELISA respectively. Out of
total 504 samples from Government farms, 4.17%, 4.17%, 3.57% and 4.36% and out of
2164 samples from private farms, 6.19%, 6.33%, 5.45% and 6.65% were positive by
RBPT, LFA, STAT and ELISA test respectively. The prevalence from 1-3 year old
was, 2.06% by RBPT and LFA, 1.28% and 2.31% by STAT and ELISA, from 3-6 year
old, 7.61%, 7.79%, 6.79% and 8.02% were positive by RBPT, LFA, STAT and ELISA
respectively and out of 572 from more than 6 year old, 2.97% by RBPT and LFA,
whereas 2.62% and 3.49% were positive by STAT and ELISA.
Out of 2484 serum samples of buffaloes, 5.68%, 5.80%, 5.43% and 6.04% were
positive by RBPT, LFA, STAT and ELISA respectively. In male buffaloes the
prevalence was 5.10% (RBPT and LFA), 3.06% (STAT) and 6.12% (ELISA). Out of
2386 females 5.69%, 5.82%, 5.53% and 6.03% were positive by RBPT, LFA, STAT
and ELISA respectively. Out of 175 samples from Government farms, 4% (RBPT and
LFA), 2.28% (STAT) and 4.57% (ELISA) were positive and out of 2309 samples from
private farms 5.80%, 5.93%, 5.67% and 6.14% were positive by RBPT, LFA, STAT
and ELISA respectively. Out of 90 samples from 1-3 year old, 3.34% were positive by
RBPT, LFA and ELISA, whereas no animal was positive by STAT, in 3-6 year old,
5.88%, 6.06%, 5.79% and 6.23% were positive by RBPT, LFA, STAT and ELISA
respectively and in more than 6 year old, 3.38% (RBPT), 2.54% (LFA and STAT) and
4.23% (ELISA) were positive.
Out of 1124 serum samples from human beings belonging to three regions,
10.68%, 10.14%, 10.05% and 11.21% were positive by RBPT, LFA, STAT and ELISA
tests respectively. Out of 900 from men 11.45%, 11.12%, 11% and 11.77%, whereas
from 224 women 7.58%, 6.25%, 6.25% and 8.92% were positive by RBPT, LFA,
STAT and ELISA respectively. The prevalence in the less than 10 years age was 5.8%
by all the four tests, in 10-20 years of age, 6.9%, 6.32%, 5.75% and 7.47%, in 20-40
years of age 12.97%, 12.35%, 12.35% and 13.53% and in more than 40 years of age,
10.03%, 9.51%, 9.51% and 10.54% were positive by RBPT, LFA, STAT and ELISA
respectively. Occupation wise the prevalence in veterinarians, 10.24% (RBPT), 9.70%
(LFA and STAT) and 10.51% (ELISA), in para veterinary staff, 11.62% (RBPT),
11.12% (LFA) 10.85% (STAT) and 12.14% (ELISA), in dairy farm workers, 11.16%
(RBPT), 10.15% (LFA and STAT), and 11.67% (ELISA), in shepherds, 10.57%
(RBPT, LFA and STAT) and 11.53% (ELISA) and in others, 6.15% (RBPT, LFA
andSTAT), and 7.69% (ELISA) were positive. Considering ELISA as gold standard test
based on its efficacy, the sensitivity and specificity were 94.12% and 100% for RBPT
and LFA, whereas for STAT 86.88% and 100% respectively.
Out of 2024 milk samples from three regions, 8.99%, 9.19% and 9.44% were
positive by MRT, LFA and ELISA respectively. The prevalence in cow milk samples
was 9.11%, 9.32% and 9.52% and in buffalo milk samples, 8.88%, 9.07% and 9.36%
by MRT, LFA and ELISA respectively. Sensitivity of MRT and LFA was found to be
of 95.28% and 97.38%, respectively, with considering m-ELISA as a gold standard test.
Out of 150 samples of serum, 14.0%, 15.33%, 13.33% and 16.67% were positive
by RBPT, LFA, STAT, ELISA respectively and in milk 12.67%, 12.0% and 13.33%
were positive by MRT, LFA and m-ELISA respectively. Sensitivity of RBPT, LFA,
STAT (Tests for serum), MRT, LFA and m-ELISA (Tests for milk) was found to be of
84%, 92%, 80%, 76%, 72% and 80%, respectively, serum ELISA as a gold standard
test, while specificity was found to be of 100% for all above tests.
Cultural, ELISA and PCR assay were compared, wherein milk samples gave
positive, 8%, 13.34% and 10.67%, in semen samples, 12%, 14% and 16%, in blood
samples, 9.33%, 16.67%, and 12% and in aborted fetuses, 40%, 50% and 45%
respectively. Higher efficacy obtained in ELISA followed PCR and culture in all the
samples except in semen samples.
Out of total 40 isolates, 29 belongs to any one of 1, 2 or 4 biovars of Br. abortus, 7
belongs to biovar 1 of Br. melitensis by AMOS PCR and 29 isolates identified as
Br.abortus, 7 as Br. melitensis, 4 as Brucella abortus Strain 19 by Bruce-ladder PCR. The
load of Brucellaorgnisms in milk ranged from 1.240 x 104CFU/ml to 1.842 x 104CFU/ml of
milk and in semen ranged from 1.362 x104 CFU/ml to 1.128 x105 CFU/ml. The sequence of
PCR products revealed that the samples tested confirming Brucella abortus.