Ragiv Gandhi Univerity of Helath Science, Karnataka Bangalore

Ragiv Gandhi Univerity of Helath Science, Karnataka Bangalore

Rajiv Gandhi Institute of Health Sciences

Karnataka, Bangalore

M. D (MICROBIOLOGY)

SriDevarajUrsMedicalCollege

Tamaka, Kolar – 563 101

SEROLOGICAL STUDY OF BRUCELLA INFECTIONS

IN THE HIGH RISK POPULATION AND IN PATIENTS

WITH FEVER IN AND AROUND KOLAR .

By

Dr.DHANALAXMI ANIYAPPANAVAR

DEPARTMENT of MICROBIOLOGY

SriDevarajUrsMedicalCollege

Tamaka, Kolar – 563 101

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES, BANGALORE,
KARNATAKA.
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECTS FORDISSERTATION
1. / Name of the candidate and Address (in block letters) / Dr DHANALAXMI.ANIYAPPANAVAR.
DEPARTMENT OF MICROBIOLOGY
SRI DEVARAJ URS MEDICAL COLLEGE,TAMAKA, KOLAR-563101
2. / Name of the institution / SRIDEVARAJURSMEDICALCOLLEGE, KOLAR- 563101
3. / Course of the study and subject / MD MICROBILOGY
4. / Date of admission to course / 31st MAY 2007
5. / Title of the topic / SEROLOGICAL STUDY OF BRUCELLA INFECTIONS IN THE HIGH RISK POPULATIONAND IN PATIENTS WITH FEVER IN AND AROUND KOLAR.
6. / Brief Resume of Intended work:
6.1 / Need for the study
Brucellosis is one of the world’s major zoonosis. It continues to be an infectious
disease of public health importance and economic concern in many parts of the
world1. The magnitude of the disease remains unknown due to the paucity of reports
and varietyof clinical manifestations2.
Brucellosis is known to be present in all livestock3. It istransmitted to human beings by consumption of raw milk, milkproducts, meat, through direct contact with infected animal parts (such as placenta by inoculation through ruptures of skin and mucous membranes),infected aerosolizedparticles and rarely from person to person2.Brucellosisis an occupational hazard4.Dairy personnel, veterinary personnel, abattoirs, animal owners, milk vendors, and shepherds, are atparticular risk.1,2
Kolar district has a large portion of its population residing in close proximity with the cattle. However there is no study doneso far on the brucella infections in Kolar area. Hence this study is undertaken.
6.2 / Review of literature
Worldwide reported incidence of brucellosis in endemic areas is estimated to vary from <0.01 to 200/100,000 population.3 It is thought that the true incidence may be 25 times higher than the reported incidence due to misdiagnosis and under-reporting.3
Brucellosis may present clinically as acute illness, chronic illness following an acute attack or chronicillness with insidious onset.2Brucellosis is known to be a multisystem disease.It may present with varied manifestations such asfever, night sweats, fatigue, joint pain, cough and breathlessness, hepatosplenomegaly, testicular pain, and scrotal swelling.2
Culture of brucella is biohazardous and diagnosis is mainly based on serology5. Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (SAT), Enzyme Linked Immunosorbent Assay (ELISA), and Complement Fixation Test (CFT) are the commonly used tests.6
Many times patients with brucellosis are labelled as Pyrexia of Undetermined Origin (PUO), and are subjected to various laboratory investigations; however these investigations usually do not include any of the serological tests for brucella.7 Thus brucella serology seems to be a very important investigation to know the magnitude of brucellosis in any region.
Rose Bengal Plate Test (RBPT) is a rapid plate test that uses suspension of brucella antigen in acid buffer.8It is able to detect agglutinating and non agglutinating antibodies and is highly sensitive. It can be used as a good screening test.8SAT and ELISA can be used as confirmatory tests.3
2Mercaptoethanol Test is a modification of Standard Tube Agglutination Test (SAT).It is performed in the laboratory similar to the SAT except for the addition of 2Mercaptoethanol (2ME) to a final concentration of 0.05M in each tube.9 It is observed that 2ME test is a better indicator of recent infection.9
ELISA with its high sensitivity and specificity is preferred overother conventional serological tests . In recent years ELISA is used for screening the population at high risk, as SAT is known tomiss many seropositives and thus may not be sensitive for thepurpose of serological screening.6 Relative to other tests the Brucella abortus ELISA was found to be highly sensitive and specific serodiagnosic test in diagnosing brucellosis in occupationally exposed workers in whom high seroprevalance is recorded6,and many of them may not have overt clinical manifestataions.
6.3 / Objectives of the study
  1. To test people at occupational risk in Kolar district for presenceof brucella infection.
  2. To detect brucella infection among patients with fever attentingR.L.JalappaHospital and Research Centre.Tamaka,Kolar, attached to SriDevarajUrsMedicalCollege, Tamaka, Kolar.

7. / Materials and Methods
7.1 / Source of data:
Serology will be performed in two groups ofpatients.
Group I: The population at risk,which will include, veterinary personnel, dairy personnel, animal owners, abattoirs, and shepherds.
Serum samples from 150 such subjects will be studied.
Group II: Patients with temperature>38.30C, of more than ten days duration
especially with history of joint pain,night sweats, cough, and
breathlessness.
One hundred such patients will be studied.
Duration of study: November 2007- March 2009.
7.2 / Method of collection of data:
/ An informed consent will be taken from the people in each group for doing the tests.
  1. 5ml venous blood will be aseptically collected from the high riskgroup people. The serum will be separated and tested for presenceof antibrucellar antibodies by RBPT and IgG ELISA.8,10.
  1. Similarly 5ml blood will be aseptically collected frompatients of all ages and both sexes, presenting with fever for evaluation. The serum will be tested for presence of antibrucellar antibodies byRBPT and SAT with 2ME.9,10.
  1. Antigen for RBPT and SATwill be obtained from Institute of Animal Health and Veterinary Biologicals. Hebbal, Bangalore.
  1. IgG ELISA kits to be used in risk population will be procured commercially.
FLOW CHART-Processing of samples
Group I Normal people exposed to brucellosis

5ml venous blood collected aseptically

Allowed to clot at room temperature

Serum separated with sterile precautions and refrigerated

Rose Bengal Plate Test IgG ELISA.
Group II Patients presenting with fever

5ml venous blood
Allowed to clot at room temperature

Serum separated with sterile precautions and refrigerated

Rose Bengal Plate Test Standard Tube Agglutination
with 2 Mercaptoethanol.
Inclusion criteria:
Group I Veterinary personnel, dairy personnel, abattoirs, shepherds, and
animal owners incontact with cattle.
Group IIPatients with temperature>38.30C, of more than ten days duration
especially with complaints of joint pain, night sweats, cough, and
breathlessness.
Exclusion criteria:
Patients with obvious causes for fever such as malaria and pulmonary tuberculosis will be excluded from the study.
Analysis:
The results will be analysed using descriptive statistics like proportions and comparision of groups will be made using Chi Square test and Z test.
7.3 / Does the study require any investigation or interventions to be conducted on patients or other humans or animals? If so, please describe.
YES. Details given in 7.2.
As required for any serological test 5ml blood will be aseptically collected from the high risk group subjects and the patients after obtaining an informed consent.
7.4 / Has ethical clearance been obtained from your institution in case of 7.3
YES, obtained. The patients and subjects diagnosed as brucellosis will be referred for appropriate treatment.
8. / List of references
  1. AgasthyaAS, Isloor S, Prabhudas K. Brucellosis In High Risk Group Individuals. Ind J Med Microbiol 2007;25:28-31.
  1. Mantur BG, Biradar MS, Bidri RC, Mulimani MS, Veerappa K, Kariholu P et al. Protean clinical manifestations and diagnostic challenges of human brucellosis in adults:16 years’experience in an endemic area. J Med Microbiol 2006;55: 897-903.
  1. Smits HL, Kadri SM. Brucellosis in India: A deceptive infectious disease. Ind J Med Res 2005; 122: 375-384.
  1. Kumar P, Singh DK, Barbuddhe SB. Seroprevalence ofBrucellosis among abattoir personnel of Delhi. J Commun Dis1997;29:131-137.
  1. Handa R, Singh S, Singh N, Wali TB. Brucellosis in North India:results of a prospective study. J Commun Dis 1998;30:85-87.
  1. Gad E I-RabMO, Kambal AM. Evaluation of a Brucella Enzyme Immunono Assay Test (ELISA) in Comparison with Bacteriological Culture and Agglutination. J Infect 1998;36:197-201.
  1. Kadri SM, Rukhsana A, Laharwal MA, Tanvir M .Seroprevalence of Brucellosis in Kashmir(India) among Patientswith Pyrexia of Unknown Origin. J Ind Med Asso 2000;98:120-121.
  1. Ruiz-Mesa JD, Gonzalez-Sanchez, Reguera JM, Martin L,Lopez-Palmero S, Colmeruro JD. Rose Bengal Test: Diagnosticyield and use for the rapid diagnosis of human Brucellosis inemergency departments in endemic areas. Clin Microbiol Infect 2005;11:221-225.
  1. Buchanan TM, Faber LC. 2 Mercaptoethanol BrucellaAgglutination Test: Usefulness for Predicting Recovery fromBrucellosis. J Clin Microbiol 1980;11:691-693.
  1. Alton GG, Jones LM, PietzPE. Laboratory Techniques InBrucellosis 2ndedn 1975.Geneva;WHO.

9. / Signature of Candidate
10. / Remarks of Guide
11. / 11.1 / Name and Designation of the Guide (In block letters) / DR. S. R. PRASAD
M.D
PROFESSOR AND HEAD DEPT. OF MICROBIOLOGY
SRI DEVARAJ URS MEDICAL
COLLEGE.TAMAKA,KOLAR.
11.2 / Signature
11.3 / Co-guide (if any) / DR. SRINIVAS. RAO
M.D.
PROFESSOR OF MEDICINE.
11.4 / Signature
11.5 / Head of the Department / Dr.S.R.PRASAD
M.D
PROFESSOR AND HEAD
11.6 / Signature
12. / 12.1 / Remarks of Chairman and Principal
12.2 / Signature
Proforma:
Name: Hosp No;
Age: Sex:
Address: Lab No:
Brief History:
H/O consumption of raw milk, milk products, meat.
H/O owning cattle, sheep.
H/O fever with joint pain, night sweats, cough and breathlessness.
Clinical Examination:
Temperature:
Investigation: Treatment:
RBPT: Outcome:
IgG ELISA:
SAT with 2ME:
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