The targets of the Structural GenomiX (SGX) bacterial genomics project were proteins conserved in multiple prokaryotic organisms with no obvious sequence homolog in the Protein Data Bank of known structures. The outcome of this work was 80 structures, covering 60 unique sequences and 49 different genes. Experimental phase determination from proteins incorporating Se-Met was carried out for 45 structures with most of the remainder solved by molecular replacement using members of the experimentally phased set as search models. An automated tool was developed to deposit these structures in the Protein Data Bank, along with the associated X-ray diffraction data (including refined experimental phases) and experimentally confirmed sequences. BLAST comparisons of the SGX structures with structures that had appeared in the Protein Data Bank over the intervening 3.5 years since the SGX target list had been compiled identified homologs for 49 of the 60 unique sequences represented by the SGX structures. This result indicates that, for bacterial structures that are relatively easy to express, purify, and crystallize, the structural coverage of gene space is proceeding rapidly. More distant sequence-structure relationships between the SGX and PDB structures were investigated using PDB-BLAST and Combinatorial Extension (CE). Only one structure, SufD, has a truly unique topology compared to all folds in the PDB.

Eleven microsatellite loci (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA023, ETH3, ETH225, BM1824) were evaluated for their use in paternity testing in the Yugoslav Pied cattle (YU Simmental cattle) population in Serbia. A total of 40 animals were tested. At the 11 tested loci, a total of 91 alleles were detected. The mean number of alleles per locus was 8.273. Polymorphism information content (PIC) values ranged from 0.58 to 0.88 with the mean value of 0.72. The most informative loci were: TGLA53 (14 alleles, PIC = 0.88), TGLA227 (11 alleles, PIC = 0.82), INRA023 (11 alleles, PIC = 0.86), BM2113 (9 alleles, PIC = 0.80). Combined power of discrimination (CPD) for the 11 microsatellite loci was 0.999. The results of the present study confirm that the analysed set of 11 microsatellite markers recommended by ISAG is suitable for paternity testing in Yugoslav Pied cattle in Serbia.

Mastitis represents a heavy burden for the dairy sector worldwide with high economic and animal welfare impact. Antibiotic treatment is an important component of mastitis control programs. However, emergence and transfer of antimicrobial-resistant (AMR) bacteria is becoming a growing concern. Therefore, the development of novel agents is required for prevention and treatment of mastitis. Hence, our aim was to assess the antibacterial properties of two essential oils (EOs) obtained from oregano (Origanum vulgare L., Lamiaceae) and mountain savory (Satureja montana L., Lamiaceae) against mastitis-associated bacteria in Serbia. The chemical composition and antioxidant potential of these EOs were also evaluated. The present study was conducted on strains derived from aseptic milk samples collected from Holstein-Friesian cows with clinical or subclinical mastitis, during the morning milking. Clinical mastitis was assessed by clinical examination, while subclinical mastitis was confirmed using somatic cell count in the milk samples. The microdilution method was used to determine the antibacterial activity, while antioxidant potential of the EOs was evaluated in several in vitro assays. The values of minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) were used to quantitatively measure the antibacterial activity of each EO. MIC/MBC ranged from 0.78/6.25 and 0.39/0.78 mg/mL for oregano and mountain savory, respectively. A total of 25 compounds were identified in the oregano EO, while 47 were identified in winter savory EO, among which aromatic oxygenated monoterpenes were the most abundant compounds. The tested EOs have shown promising antimicrobial activity and could be considered as one of the treatment approaches in mastitis-affected cows.

Leptin, a biomolecule secreted by adipose tissue, enchances productivity in cattle, especially affecting milk traits. The aim of this study was to detect leptin gene polymorphism on exon 3 (A59V locus) and intron 2 (SAU3AI locus) in the endangered population of autochtonous Busha cattle and associations with milk traits. The study included 46 cows: 36 Busha and 10 half-bred. Milk analyses comprised determination of somatic cell counts, fat, protein, lactose, total solids and solids-not-fat (SNF) concentrations and freezing point depression (FPD). Polymorphisms were determined by PCR-RFLP technique. A single A59V genotype (CC) was affirmed, and two SAU3AI genotypes, AA and AB, with frequencies of 78.26% and 21.74%, respectively. Comparing the obtained results for chemical characteristics of milk between cows with AA and AB, no significant differences were found, except for SNF content and FPD values. Cows with AA genotype had significantly lower (p=0.021) average SNF content (8.74%) in milk compared to the average SNF content (9.28%) in those with genotype AB, while cows with genotype AA (−0.54°C) had significantly higher (p=0.004) average FPD values than those with AB genotype (−0.58°C). The absence of BB genotype and significant differences in the investigated functional traits between two SAU3AI genotypes and the absence of A59V polymorphism (presence of only CC genotype) show that the Busha cattle breed, although being an autochtonous low-producing native breed used for meat and milk production, harbours polymorphism on gentic markers characteristic of high production dairy cows.

The aim of this study was to determine the distribution of κ-casein (κ-CN) and β-lactoglobulin (β-Lg) genotypes in the autochtonous (Busha) and dairy (Holstein-Friesian, HF) cattle breeds with PCR-RFLP (polymerase chain reaction -restriction fragment length polymorphism). For the amplification of κ-CN and β-Lg gene fragments specific primers were used. After digestion with specific endonucleases genotypes were determined for both genes in 18 Busha and 19 HF cows. The results showed that κ-CN gene was represented with the AA genotype in 31.58 % HF cows, AB in 52.63 % cows, whilst the genotype BB was found in 15.79 % cows only. Among the examined Busha cattle 44.44 % cows had AA genotype and 55.56 % genotype AB for κ-CN. As for β-Lg gene in HF breed, AA genotype was found in 26.31 % cows, AB in 63.16 % and BB in 10.53 % cows. In Busha cows the following genotypes were established for β-Lg gene: AA in 44.44 % cows and AB in 55.56 %, whilst BB genotype was not found. These results indicate that Busha cows had a higher presence of A allelic forms of both genes (k-CN and β-lactoglobulin) than HF cows.