Brucella spp. distribution, hosting ruminants from Greece, applying various molecular identification techniques

Area of Science:

• Veterinary Microbiology
• Epidemiology
• Molecular Diagnostics

Background:

• Brucellosis is an endemic zoonotic disease in Greece, impacting livestock and human health.
• Ruminants are natural hosts, and human transmission poses occupational risks.
• Accurate Brucella species distribution data is crucial for disease control.

Purpose of the Study:

• To establish baseline Brucella genotyping data in Greece at species and subtype levels.
• To evaluate and compare different molecular techniques for Brucella detection.
• To assess the efficacy of 16S rRNA, BP26, OMP31 sequencing, and MLVA typing.

Main Methods:

• Analysis of 39 tissue samples from 30 domestic ruminants positive for Brucella via screening PCR.
• Application of four molecular techniques: 16S rRNA, BP26, and OMP31 gene sequencing, and MLVA typing.
• Comparison of results for Brucella species identification and subtyping.

Main Results:

• 16S rRNA sequencing identified Brucella only at the genus level.
• BP26 sequencing and MLVA typing accurately identified B. melitensis and B. abortus.
• OMP31 sequencing failed to yield clear results.
• BP26 and MLVA showed agreement, with BP26 being effective even with lower quality DNA.

Conclusions:

• Brucella bacteria are consistently circulating in Greek ruminant populations.
• BP26 gene sequencing is a highly effective alternative to MLVA typing for Brucella identification.
• BP26 sequencing offers advantages, especially when dealing with degraded DNA samples.