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Sero-Epidemiology Of Johne’s Disease Along With Its Effect On Serum Biochemical Profile In Cattle In District D.I. Khan, Khyber Pakhtunkhwa

By: Nasrullah (2009-VA-90) | Prof. Dr. Aneela Zameer Durrani.
Contributor(s): Dr. Waseem Yaqub | Dr. Mamoona Chaudhry.
Material type: materialTypeLabelBookPublisher: 2016Description: 49p.Subject(s): Department of Clinical Medicine and Surgery | CMS | Clinical MedicineDDC classification: 2526-T Dissertation note: Johne’s disease (JD) or Paratuberculosis caused by Mycobacterium avium subsp. Paratuberculosis (MAP) is characterized by an incubation period of several years. It is a chronic non-treatable disease of animals causing enteritis which ultimately leads to economic losses to dairy and meat industries worldwide. To estimate sero-epidemiology of the JD through ELISA is inexpensive, efficient, single step and sensitive for the detection of antibodies in cattle and buffaloes. So, keeping in view the economic and public health significance of the disease, the present study was designed with the objective to estimate the sero prevalence of JD using indirect ELISA and to quantify the effect of JD on the level of serum proteins in cattle of D. I. Khan district, Khyber Pakhtunkhwa Province. Johne’s disease is prevalent in cattle of D I Khan that affects serum proteins level in dairy animals. For the study animals were well restrained and about 5 ml blood was collected in Non- EDTA coated vacutainer from the jugular vein of each animal using 5 ml disposable syringe, after collection and proper labeling samples were transferred in Ice packed cooler to Microbiology laboratory, Govt. College of Veterinary Sciences D.I Khan. Serum was collected at the top of the vacutainer with the help of disposable pipette and was stored in eppendorf tubes in deep freezer until transported to University of Veterinary and Animal Sciences, Lahore for further analysis. Data was captured on questionnaires about the clinical signs and symptoms, management practices and possible risk factors associated with Johne’s disease, was obtained from the owners. Summary 40 The collected serum samples were analyzed by Indirect ELISA using commercial ELISA kit (Mycobacterium Paratuberculosis Antibody Test Kit, IDEXX, USA) according to manufacturer instructions. The ELISA positive samples were processed further for the estimation of serum protein analysis using Biuret method (Doumas et al. 1981;Spencer and Price. 1977).The serum albumin was determined by Photometric colorimetric test (Bromocresole green (BCG) method (Spencer and Price. 1977).The globulin will be estimated by subtracting serum albumin from serum total proteins (Javed et al. 2010). Data regarding sero-prevalence by indirect ELISA were analyzed through Pearson's Chisquare Test, while data regarding the effect on serum protein was analyzed by simple t-test using statistical package for social sciences (SPSS) version 20. P < .05 was considered significant. The present study demonstrated (12.7%) prevalence of MAP in cattle population of D.I Khan. There were no significant effect of MAP on serum biochemical profile. The results of all variables studied as under breed, gender, age, vaccinated status and management system. The most frequent level was observed in females 13.7% as compared to males 0% (0/4). Among different age groups, higher sero-prevalence 33.3% were present at age group above 6 years while sero-prevalence of 7.1% in 2-3 years and 0% were recorded in age group 4-6 years. Statistically a significant different were present between these groups. Moreover, there was no relationship of vaccination status with disease because the prevalence recorded 16.6% and vaccinated cases positive were recorded 0% but there was no significance difference among two groups of animals. Breed wise risk factor statistically significant because the MAP cases were highly prevalent in non-descript cattle 25% as compared to crossbred 5.8% and Sahiwal 0% cattle breed. BCS category I had highest prevalence 66.6% while in BCS category II prevalence Summary 41 was 23.8% and in BCS category III the prevalence was 0% recorded. Statistically the BCS of the disease animals was significantly associated. Highest prevalence 100% were recorded above 60 days of diarrhea duration while 0% were recorded less than 60 days of diarrhea duration. Statistically the sero-prevalence of diarrhea duration was highly significant. Management wise sero-prevalence of Johne’s disease on the type of housing was higher in open 19.2% as compared to confined 6.8% showing non-significant difference statistically. Sanitation wise seroprevalence were higher in poor 19.2% as compared to good sanitation 6.8% showing also nonsignificant difference statistically. Sero-prevalence of disease was less in well water 0% as compared to tap water 25% showing there were a significant difference between the seroprevalence of Johne’s disease under different water conditions. The results of total protein analysis was statistically non-significant.
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Johne’s disease (JD) or Paratuberculosis caused by Mycobacterium avium subsp.
Paratuberculosis (MAP) is characterized by an incubation period of several years. It is a chronic
non-treatable disease of animals causing enteritis which ultimately leads to economic losses to
dairy and meat industries worldwide. To estimate sero-epidemiology of the JD through ELISA is
inexpensive, efficient, single step and sensitive for the detection of antibodies in cattle and
buffaloes. So, keeping in view the economic and public health significance of the disease, the
present study was designed with the objective to estimate the sero prevalence of JD using
indirect ELISA and to quantify the effect of JD on the level of serum proteins in cattle of D. I.
Khan district, Khyber Pakhtunkhwa Province. Johne’s disease is prevalent in cattle of D I Khan
that affects serum proteins level in dairy animals.
For the study animals were well restrained and about 5 ml blood was collected in Non-
EDTA coated vacutainer from the jugular vein of each animal using 5 ml disposable syringe,
after collection and proper labeling samples were transferred in Ice packed cooler to
Microbiology laboratory, Govt. College of Veterinary Sciences D.I Khan. Serum was collected
at the top of the vacutainer with the help of disposable pipette and was stored in eppendorf tubes
in deep freezer until transported to University of Veterinary and Animal Sciences, Lahore for
further analysis. Data was captured on questionnaires about the clinical signs and symptoms,
management practices and possible risk factors associated with Johne’s disease, was obtained
from the owners.
Summary
40
The collected serum samples were analyzed by Indirect ELISA using commercial ELISA
kit (Mycobacterium Paratuberculosis Antibody Test Kit, IDEXX, USA) according to
manufacturer instructions.
The ELISA positive samples were processed further for the estimation of serum protein analysis
using Biuret method (Doumas et al. 1981;Spencer and Price. 1977).The serum albumin was
determined by Photometric colorimetric test (Bromocresole green (BCG) method (Spencer and
Price. 1977).The globulin will be estimated by subtracting serum albumin from serum total
proteins (Javed et al. 2010).
Data regarding sero-prevalence by indirect ELISA were analyzed through Pearson's Chisquare
Test, while data regarding the effect on serum protein was analyzed by simple t-test using
statistical package for social sciences (SPSS) version 20. P < .05 was considered significant.
The present study demonstrated (12.7%) prevalence of MAP in cattle population of D.I
Khan. There were no significant effect of MAP on serum biochemical profile. The results of all
variables studied as under breed, gender, age, vaccinated status and management system. The
most frequent level was observed in females 13.7% as compared to males 0% (0/4). Among
different age groups, higher sero-prevalence 33.3% were present at age group above 6 years
while sero-prevalence of 7.1% in 2-3 years and 0% were recorded in age group 4-6 years.
Statistically a significant different were present between these groups. Moreover, there was no
relationship of vaccination status with disease because the prevalence recorded 16.6% and
vaccinated cases positive were recorded 0% but there was no significance difference among two
groups of animals. Breed wise risk factor statistically significant because the MAP cases were
highly prevalent in non-descript cattle 25% as compared to crossbred 5.8% and Sahiwal 0%
cattle breed. BCS category I had highest prevalence 66.6% while in BCS category II prevalence
Summary
41
was 23.8% and in BCS category III the prevalence was 0% recorded. Statistically the BCS of the
disease animals was significantly associated. Highest prevalence 100% were recorded above 60
days of diarrhea duration while 0% were recorded less than 60 days of diarrhea duration.
Statistically the sero-prevalence of diarrhea duration was highly significant. Management wise
sero-prevalence of Johne’s disease on the type of housing was higher in open 19.2% as compared
to confined 6.8% showing non-significant difference statistically. Sanitation wise seroprevalence
were higher in poor 19.2% as compared to good sanitation 6.8% showing also nonsignificant
difference statistically. Sero-prevalence of disease was less in well water 0% as
compared to tap water 25% showing there were a significant difference between the seroprevalence
of Johne’s disease under different water conditions. The results of total protein
analysis was statistically non-significant.

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