Investigation of a Membrane Filter Protocol for Direct Identification of Microorganisms in Dog Ear Discharge using MALDI-TOF MS

Otitis externa is an ear disease that is present in 10-20% of dogs. Bacterial organisms are a common cause, and infection may cause severe and chronic inflammation. Thus, there is a need for early detection and identification of bacteria in ear discharge samples. In this study, 31 such samples were analyzed to identify microorganisms using MALDI-TOF MS with pretreatment using a membrane filtration protocol and a rapid BACpro II kit, compared to pretreatment with a rapid BACpro II kit alone. Reliable scores for E. coli and E. faecalis were obtained after inoculation of samples with 1.0×10⁴ CFU/ml using the membrane filter with the rapid BACpro II kit, with approximately 10-fold higher sensitivity compared to rapid BACpro pretreatment alone. Among 31 bacteria-positive colonies in the samples, MALDI-TOF MS and membrane filtration (0.80 µm) with a rapid BACpro II kit was significantly more accurate compared to use of the kit alone (29 (82.9%) vs. 17 (48.6%) isolates identified, p<0.001). These results show that MALDI-TOF MS with a membrane filter and a rapid BACpro II kit is a quick and reliable method for bacterial identification in ear discharge samples.