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CORRESPONDENCE
Year : 2014  |  Volume : 139  |  Issue : 5  |  Page : 776-778

Rapid detection of Brucella by an automated blood culture system at a tertiary care hospital of north India


Department of Medical Microbiology, Postgraduate Institute of Medical Education & Research, Chandigarh 160 012, India

Date of Web Publication9-Jul-2014

Correspondence Address:
Pallab Ray
Department of Medical Microbiology, Postgraduate Institute of Medical Education & Research, Chandigarh 160 012
India
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Source of Support: None, Conflict of Interest: None


PMID: 25027090

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How to cite this article:
Raj A, Gautam V, Gupta PK, Sethi S, Rana S, Ray P. Rapid detection of Brucella by an automated blood culture system at a tertiary care hospital of north India. Indian J Med Res 2014;139:776-8

How to cite this URL:
Raj A, Gautam V, Gupta PK, Sethi S, Rana S, Ray P. Rapid detection of Brucella by an automated blood culture system at a tertiary care hospital of north India. Indian J Med Res [serial online] 2014 [cited 2020 Sep 22];139:776-8. Available from: http://www.ijmr.org.in/text.asp?2014/139/5/776/136472

Sir,

Brucellosis is a significant veterinary and public health problem in endemic areas including the Mediterranean countries, the Middle East, the African subcontinent, the Latin America and parts of Asia [1] . Species prevalent in India include Brucella melitensis and B. abortus[2]. B. melitensis is the most virulent and the commonest species and it causes severe and prolonged disease with a risk of disability in man. Goats, reared for meat, constitute the main source of infection. Goat and sheep milk is used for adulteration when there is a shortage of cows and buffalos milk in summer months [3] . B. abortus is the main species in cattle [4] , and bovine brucellosis is widespread in India [5] . Brucella species are capable of evading host defense mechanisms, surviving as intracellular organisms, and are able to cause prolonged morbidity, relapses, and long-term sequelae. It has very low mortality (1%) but high morbidity [6] . It may affect any organ of the body with clinical manifestations that include fever, joint pains, loss of weight, sweating, cough, sciatica, splenic enlargement, liver enlargement, orchitis, etc[7] . The non-specific and protean clinical presentation mimics other infectious and non-infectious conditions and consequently, the diagnosis of the disease is often missed or delayed [8] .

The present study was conducted in a tertiary care hospital in Chandigarh, north India. The culture confirmed cases of Brucella infection admitted to or attending various outpatient clinics of the Post-graduate Institute of Medical Education and Research (PGIMER), Chandigarh, during March 2010 to June 2012 were retrospectively studied. Blood culture was performed in the clinical bacteriology laboratory in the department of Medical Microbiology as a part of routine diagnostic services using BACTEC 9240 (BD Diagnostics, USA) system. Standard aerobic/F and Peds plus/F media (BD Diagnostics, USA) were used for the adult and the paediatric patients, respectively. Inoculated bottles were monitored up to five days or until those became positive [9] . The flagged positive vials were subjected to Gram stained smear microscopy and subcultured on sheep blood agar and MacConkey agar, incubated at 37°C. Brucella was identified and differentiated from other Gram negative genera on the basis of small, translucent, soft and easily emulsifiable colonies on MacConkey and blood agar (non-pigmented and non-haemolytic) with absence of X and V factor dependence; Gram-negative tiny coccobacilli, non-encapsulated, non-motile, oxidase, catalase and urease positive, producing acid from xylose in oxidative fermentative medium [6] . The results of culture were compared with standard agglutination tube test (SAT) for Brucella with or without pre-treatment of serum with 0.05M 2-mercaptoethanol (2-ME) using antigen obtained from Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh. During the study period, Brucella spp. was isolated from 13 blood and one CSF specimens culture. Clinical details were available in seven cases. The patients in general, had one to three weeks history of fever associated with chills and rigour, malaise, anorexia, headache and vomiting. Two patients had hepatosplenomegaly and three had only hepatomegaly and history of weight loss. Associated risk factors includes intake of nonpasteurized milk or milk products, rearing cattle, veterinarian as profession, and laboratory exposure. In all the cases Brucella serology using SAT without 2ME pre-treatment demonstrated a titre of 2560 IU. The quantity of specific IgG as determined by treatment of serum with 2-ME was 2560 IU in six cases and 1280 IU in one case [Table 1].
Table 1: Clinical profile and outcome of confirmed cases with Brucella infection


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The only confirmatory evidence of brucellosis is the recovery of the bacterium from the patient. Therefore, culture is considered the gold standard in the laboratory diagnosis of brucellosis [6] . Various blood culture methods are available but the newer semiautomatic methods such as BACTEC 9240 has shortened the time taken for detection; the presence of Brucella can be detected with these methods within five days of incubation [10] . In the present study, nine cultures became positive by the third day and all were detected within five days. In one patient, blood and CSF samples were received in BACTEC bottles and both were positive on the third day (though CSF inoculation in these vials is not routinely recommended) while the same CSF was negative by conventional culture. Direct invasion of the central nervous system occurs in less than 5 per cent cases of brucellosis [11] . Therefore, rapid detection of scanty load of bacteria in CSF by automated culture system is an added advantage for sterile body fluids.

All the proven cases received proper treatment [11],[12] because of rapid and accurate aetiological diagnosis aided by use of automated blood culture system, and recovered.

 
   References Top

1.Smits HL, Kadri SM. Brucellosis in India: a deceptive infectious disease. Indian J Med Res 2005; 122 : 375-84.  Back to cited text no. 1
    
2.Mathur TN. An outbreak of brucellosis in Bhiwani. The danger of infection with Brucella melitensis. Indian J Med Sci 1962; 16 : 878-80.  Back to cited text no. 2
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3.Mathur TN. The role of the goat in human brucellosis in India with particular reference to infection with Brucella melitensis. J Assoc Physicians India 1964; 12 : 805-13.  Back to cited text no. 3
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4.Mantur BG, Amarnath SK. Brucellosis in India - a review. J Biosci 2008; 33 : 539-47.  Back to cited text no. 4
    
5.Renukaradhya GJ, Isloor S, Rajasekhar M. Epidemiology, zoonotic aspects, vaccination and control/eradication of brucellosis in India. Vet Microbiol 2002; 90 : 183-95.  Back to cited text no. 5
    
6.Lindquist D, Chu MC, Probert WS, Francisella and Brucella. In: Murray PR, Baron EJ, Landry ML, Jorgensen JH, Pfaller MA, editors. Manual of clinical microbiology, 9 th ed. Washington DC: American Society of Microbiol; 2007. p. 815-34.  Back to cited text no. 6
    
7.Mathur TN. A study of human brucellosis based on cultures isolated from man and animals. Indian J Med Res 1968; 56 : 250-8.  Back to cited text no. 7
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8.Lulu AR, Araj GF, Khateeb MI, Mustafa MY, Yusuf AR, Fenech FF. Human brucellosis in Kuwait: a prospective study of 400 cases. Q J Med 1988; 66 : 39-54.  Back to cited text no. 8
    
9.Clinical and Laboratory Standards Institute (CLSI). Principles and procedures for blood cultures; Approved guideline: vol. 27, M47-A. Wayne, PA, USA: CLSI ; 2007.  Back to cited text no. 9
    
10.Bannatyne RM, Jackson MC, Memish Z. Rapid diagnosis of Brucella bacteremia by using the BACTEC 9240 system. J Clin Microbiol 1997; 35 : 2673-4.  Back to cited text no. 10
    
11.Young EJ. Brucella species. In: Mandell GL, Bennett JE, Dolin R, editors. Principles and practice of infectious diseases, 7 th ed. Philadelphia: Churchill Livingstone; 2010. p. 2921-6.  Back to cited text no. 11
    
12.Salvatore M, Meyers BR. Tetracylcines and chloramphenicol. In: Mandell GL, Bennett JE, Dolin R, editors. Principles and practice of infectious diseases, 7 th ed. Philadelphia: Churchill Livingstone; 2010. p. 385-401.  Back to cited text no. 12
    



 
 
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