Protein Based in Vivo Diagnostics for Endometrial Biofilms

Summary: Successful management of bacterial biofilms in the uterus requires an accurate diagnostic in vivo assay that we propose to develop. Scientific Importance: Bacteria within a biofilm are protected from the immune system and are not responsive to standard antibiotic treatments. This can contribute to the development of bacterial resistance against antibiotics, which is a global health issue in both animals and people. It is therefore, important to accompany any antibiotic with treatment that disrupts bacterial biofilms in order to prevent the development of antibiotic resistance. However, there is currently no available assay to diagnose the condition in the uterus. While current diagnostics can provide information on if the bacteria of interest is capable of developing a biofilm under optimal conditions, it leaves the clinician guessing if the case that is presented to her/him actually is associated with biofilm formation. The development of an accurate in vivo assay to diagnose the condition is therefore necessary. Based on current literature, we believe that uterine biofilm forming bacteria produce and secrete specific proteins that can be used to diagnose the condition. Here we propose to develop a protein based diagnostic test for bacterial biofilms in the equine uterus. - First, we propose to identify secreted proteins that are associated with bacterial biofilms from the most common bacteria associated with equine endometritis. - Second, we will induce bacterial biofilms from these bacteria in the uterus, and explore methods to detect proteins of interest in uterine lavage fluid from the infected mares. Importance to the Industry: Endometritis is the most common cause of infertility in the mare, and ranked as one of the top overall medical problems in horses. Infectious endometritis is routinely treated with appropriate antibiotics, and a failure to respond to this treatment may be due to the formation of bacterial biofilms in the uterus that requires additional treatment strategies. A failure to manage bacterial biofilm properly can eventually result in bacterial antibiotic resistance, and it is necessary to rely on accurate diagnostics for the condition before a treatment plan is developed. However, there is no clinical tests available to diagnose bacterial biofilms in affected mares. Therefore, we propose to develop a diagnostic test to detect bacterial biofilms in the uterus. The successful outcome of this project will not only improve fertility of valuable bloodstock, but also contribute to public health by optimizing and reducing inappropriate use of antibiotics in equine veterinary gynecology.