Evaluation of Three Formulations of Culture Media for Isolation of<i>Brucella</i>spp. regarding Their Ability to Inhibit the Growth of Contaminating Organisms

Vicente, Acácia Ferreira; Antunes, João Marcelo Azevedo de Paula; Lara, Gustavo Henrique Batista; Mioni, Mateus Souza Ribeiro; Allendorf, Susan Dora; Peres, Marina Gea; Appolinário, Camila Michele; Listoni, Fernando José Paganini; Ribeiro, Márcio Garcia; Megid, Jane

doi:10.1155/2014/702072

Cited by 17 publications

(22 citation statements)

References 9 publications

(14 reference statements)

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“…Additionally, this method poses a risk to laboratory personnel and requires specific biosafety measures [22]. Hence, culture and biochemical typing remain the "gold standard" for the diagnosis of Brucella infection [23], including biochemical tests like CO 2 requirement, H 2 S production, and dye sensitivity. Urease, oxidase, and catalase tests are also used for the typing of Brucella spp.…”

Section: Introductionmentioning

confidence: 99%

Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt

et al. 2019

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Brucellosis is a highly contagious zoonosis worldwide with economic and public health impacts. The aim of the present study was to identify Brucella (B.) spp. isolated from animal populations located in different districts of Egypt and to determine their antimicrobial resistance. In total, 34-suspected Brucella isolates were recovered from lymph nodes, milk, and fetal abomasal contents of infected cattle, buffaloes, sheep, and goats from nine districts in Egypt. The isolates were identified by microbiological methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Differentiation and genotyping were confirmed using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis (AMOS) and Bruce-ladder PCR. Antimicrobial susceptibility testing against clinically used antimicrobial agents (chloramphenicol, ciprofloxacin, erythromycin, gentamicin, imipenem, rifampicin, streptomycin, and tetracycline) was performed using E-Test. The antimicrobial resistance-associated genes and mutations in Brucella isolates were confirmed using molecular tools. In total, 29 Brucella isolates (eight B. abortus biovar 1 and 21 B. melitensis biovar 3) were identified and typed. The resistance of B. melitensis to ciprofloxacin, erythromycin, imipenem, rifampicin, and streptomycin were 76.2%, 19.0%, 76.2%, 66.7%, and 4.8%, respectively. Whereas, 25.0%, 87.5%, 25.0%, and 37.5% of B. abortus were resistant to ciprofloxacin, erythromycin, imipenem, and rifampicin, respectively. Mutations in the rpoB gene associated with rifampicin resistance were identified in all phenotypically resistant isolates. Mutations in gyrA and gyrB genes associated with ciprofloxacin resistance were identified in four phenotypically resistant isolates of B. melitensis. This is the first study highlighting the antimicrobial resistance in Brucella isolated from different animal species in Egypt. Mutations detected in genes associated with antimicrobial resistance unravel the molecular mechanisms of resistance in Brucella isolates from Egypt. The mutations in the rpoB gene in phenotypically resistant B. abortus isolates in this study were reported for the first time in Egypt.

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Section: Introductionmentioning

confidence: 99%

Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt

et al. 2019

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“…As a result of these developments, there are a lot of selective media with different basal media, antibiotic mixtures, and concentrations (15). It was emphasized that each medium has a particular effect on the species and biovar of the genus Brucella and contaminants on account of these differences (16,17).…”

Section: Introductionmentioning

confidence: 99%

Comparison of the isolation and inhibition abilities ofselective media used for Brucella spp. Isolation

Karagül¹,

İkiz²

2017

Turk J Vet Anim Sci

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The aim of this study was to compare and contrast the isolation ability of selective media developed for Brucella spp. isolation and their inhibition ability against contaminant microorganisms. Fifty-one field strains biotyped from abortion case samples and 25 Brucella spp. negative organ samples were used. Strain suspensions and organ suspensions were prepared separately. The turbidity of the strain suspensions was measured via spectrophotometer. The bacterial density of the strain suspensions was prepared in such a way that the suspensions would have different turbidity values. Inoculation into Farrell, Agrifood Research and Technology Center of Aragon, Jones & Morgan, and Modified Thayer Martin media was performed simultaneously from a dilution of each strain suspension and organ suspension. It was incubated in a 37 °C, 5%-10% CO 2 condition for 5-8 days. Brucella isolates were identified via a conventional biotyping method. The results of this study illustrate that selective media have isolation and cultivation ability for B. abortus biovar (bv) 3, B. abortus bv1, B. melitensis bv3, B. melitensis bv1, Rev1, and S-19 strains isolated from the field. There is more variation in the contaminant inhibition ability of the media compared to their isolation sensitivity. Farrell medium has 47 Brucella spp. isolations in 51 samples. It showed the highest performance with an isolation sensitivity of 92.1% and an inhibition ability of 80%. In this context, it might be suggested that researchers should initially use Farrell medium and, afterwards, use it simultaneously with another medium to be able to increase the isolation ability.

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“…These factors make brucellosis an urgent problem. Research and development of molecular marked vaccines is an inevitable tendency of research and application of new Brucella vaccines (Vicente et al, 2014). The search for specific protein antigens has gained wide attention.…”

Section: Discussionmentioning

confidence: 99%

“…The splenocytes and myeloma cells Sp2/0 of the immunized mice were fused using the polyethylene glycol method. Screened positive hybridoma cells were cloned using the limiting dilution method (Vicente et al, 2014). The specific resistance of the hybridoma cell strains obtained was detected by ELISA.…”

Section: Preparation and Identification Of Omp31 Mabmentioning

confidence: 99%

Immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody

Zheng¹,

Wang²,

Zhang³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. We examined the immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody. Genomic DNA from the M5 strain of goat Brucella was amplified by polymerase chain reaction and cloned into the prokaryotic expression vector pGEX-4T-1. The expression and immunological characteristics of the fusion protein GST-omp31 were subjected to preliminary western blot detection with goat Brucella rabbit immune serum. The Brucella immunized BALB/c mouse serum was detected using purified protein. The high-potency mouse splenocytes and myeloma Sp2/0 cells were fused. Positive clones were screened by enzyme-linked immunosorbent assay to establish a hybridoma cell line. Mice were inoculated intraperitoneally with hybridoma cells to prepare ascites. The mAb was purified using the n-caprylic acidammonium sulfate method. The characteristics of mAb were examined using western blotting and enzyme-linked immunosorbent assay. A 680-base pair band was observed after polymerase chain reaction. Enzyme digestion identification and sequencing showed that the pGEX-4T-1-omp31 11965-11974 (2015) prokaryotic expression vector was successfully established; a target band of approximately 57 kDa with an apparent molecular weight consistent with the size of the target fusion protein. At 25°C, the expression of soluble expression increased significantly; the fusion protein GST-omp31 was detected by western blotting. Anti-omp31 protein mAb was obtained from 2 strains of Brucella. The antibody showed strong specificity and sensitivity and did not cross-react with Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, or Bacillus pyocyaneus. The pGEX-4T-1-omp31 prokaryotic expression vector was successfully established and showed good immunogenicity. The antibody also showed strong specificity and good sensitivity.

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Evaluation of Three Formulations of Culture Media for Isolation ofBrucellaspp. regarding Their Ability to Inhibit the Growth of Contaminating Organisms

Cited by 17 publications

References 9 publications

Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt

Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt

Comparison of the isolation and inhibition abilities ofselective media used for Brucella spp. Isolation

Immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody

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