Macrococcus animalis sp. nov. and Macrococcus equi sp. nov., isolated from different animals' origins.

Nine Gram-positive, non-motile, facultatively anaerobic cocci, designated EM39Eᵀ, JEK85, 18KM676, 21M1142, 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573, were isolated from diverse animal material, including horse and pig skin, bovine mastitis milk and feline urine from a urinary tract infection. Phylogenomic analysis based on amino-acid alignment obtained from translated whole-genome sequences; digital DNA-DNA hybridization (dDDH); 16S rRNA, dnaJ, hsp60, rpoB and sodA gene comparison; and MALDI-TOF MS spectral profiles placed the strains within the Macrococcus caseolyticus clade. They were closely related to Macrococcus bohemicus CCM 7100ᵀ, Macrococcus epidermidis CCM 7099ᵀ, Macrococcus goetzii CCM 4927ᵀ and Macrococcus capreoli DSM 113939^T. The novel species exhibited 22.30% dDDH and 79.49% average nucleotide identity (ANI) to each other, <24% dDDH and <82% ANI to their closest relatives, values below established species delineation thresholds, confirming their classification as two distinct novel species. Chemotaxonomic analysis identified, in both species, diphosphatidylglycerol, phosphatidylglycerol, aminolipid, glycolipids and phosphoglycolipids, as the major polar lipids. Fatty acid profiling revealed C_14 : 0, C_16 : 1 ω11c, C_16 : 0 and C_18 : 1 ω13c as predominant components, with antesio-C_15 : 0 in one species. The peptidoglycan type was A3α l-Lys-Gly2. The novel species can be distinguished from their closest relatives by their ability to grow in 12.5% NaCl. EM39Eᵀ, JEK85, 18KM676 and 21M1142 can be distinguished by positive reaction to d-ribose, and 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573 by the absence of α-glucosidase activity and acid production from methyl-β-d-glucopyranoside and maltose. The two novel species can be differentiated from each other by d-ribose metabolism and methyl-β-d-glucopyranoside hydrolysis. 18KM445^T, 18KM444, 18KM245, 18KM583 and 21KM1573 formed a distinct cluster of strains, designated Macrococcus equi sp. nov., with strain 18KM445ᵀ (=DSM 118744ᵀ; =CCOS 2124ᵀ; =CCM 9438ᵀ) as the proposed type strain. The second cluster, containing strains EM39Eᵀ, JEK85, 18KM676 and 21M1142, was designated Macrococcus animalis sp. nov., with strain EM39Eᵀ (=DSM 118743ᵀ; =CCOS 2125ᵀ; =CCM 9439ᵀ) as the proposed type strain.