MLVA-16 analysis
MLVA-16 including Panel 1 (bruce06, bruce08, bruce11, bruce12, bruce42, bruce43, bruce45, and bruce55) and 8 microsatellite markers including panel 2A (bruce18, bruce19, and bruce21), and panel 2B (bruce04, bruce07, bruce09, bruce16, and bruce30) (Le Fleche et al., 2006; Al Dahouk et al. , 2007) were performed for the B. melitensis bv3 isolates (n=41). In phylogeny, dendrograms were performed after uploading the VNTRs data and estimating theBrucella genotypes online through the MLVA bank for microbe genotyping (http://microbesgenotyping.i2bc.paris-saclay.fr). Dendrograms seeking the genetic similarities among the 41Brucella strains were based on the categorical coefficient with distance calculation and unweighted pair group method with arithmetic mean (UPGMA) using BioNumerics version 7.6 (Applied Maths, Belgium).
VNTRs data of the local B. melitensis  strains used in this study were compared with 118 B. melitensis  strains recovered from different animal species and humans from other Egyptian governorates (Sayour et al. 2020). The standard minimum spanning tree (MST), based on categorical coefficient with double locus variance priority rules as well as the dendrogram of the supplementary file 1, was used to study the genetic similarities between the local strains along with the MLVA-16 global metadata of the B. melitensis  bv3 strains (n=358) isolated from selected African countries (neighborhood) and worldwide. The genetic diversity of each MLVA-16 loci was estimated using the HGDI with 95% confidence intervals through the V-DICE tool available at the HPA website (http://www.hpa-bioinformatics.org.uk/cgi-bin/DICI/DICI.pl) where it ranged from 0 (identical strains) to 1 (different strains) as reported by Hunter and Gaston (1988). Sola et al. (2003) have classified the allelic diversity (HGDI) as high if the discriminatory power of HGDI is more than 0.6, moderate discrimination if 0.3 ≤ HGDI≤0.6, and poor discrimination if HGDI < 0.3.