| Summary: | Infectious causes of myositis are reported relatively uncommonly in horses. Among them, bacterial causes include <i>Streptococcus equi</i> subsp. <i>zooepidemicus</i>, <i>Actinobacillus equuli</i>, <i>Fusobacterium</i> spp. <i>Staphylococcus</i> spp, and <i>Corynebacterium pseudotuberculosis</i>. Infection can be spread to muscles via haematogenous or extension from skin lesions. Parasitic myositis has also been documented. In this report, a 12 year-old Italian Quarter Horse mare presented with diffuse subcutaneous nodules and masses ranging from 2 × 3 to 5 × 20 cm in size, and adherent to subcutis and muscles that were first macroscopically and cytologically diagnosed as pyogranulomas. Subsequently, histological, molecular, bacteriological, and biochemical investigations were performed. All the data obtained allowed to diagnose a severe and diffuse multibacterial granulomatous myositis caused by <i>Corynebacterium pseudotuberculosis</i> and <i>Corynebacterium amycolatum</i>. Following the therapy and an initial disappearance of most of the lesions together with a general improvement of the mare, the clinical condition deteriorated, and new nodules appeared. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and PCR techniques revealed the presence of bacteria as <i>Glutamicibacter creatinolyticus</i> and <i>Dietzia</i> spp. To the authors’ knowledge, this case report represents the first description of multibacterial granulomatous myositis due to <i>Corynebacterium pseudotuberculosis</i>, <i>Corynebacterium amycolatum</i>, <i>Glutamicibacter creatinolyticus</i>, and <i>Dietzia</i> spp. in a horse reared in Italy.
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