The aim of this study was to propose the use of a new rapid and user-friendly diagnostic tool for the detection of Macrorhabdus ornithogaster infection in birds. The current report focuses on the diagnostic feasibility of different methods, with particular emphasis on the application of the mini-Flotac technique for the diagnosis of M. ornithogaster infection. The mini-Flotac method is particularly tailored for epidemiological monitoring and surveillance, where large numbers of fecal samples must be rapidly, yet reliably, examined. Gram staining, as the standard method, was used to validate the reliability of the mini-Flotac method. This tool has not yet been used in avian species or in the diagnosis of yeast infections. In our study, M. ornithogaster showed excellent performance in a flotation assay, which had not been demonstrated previously. Our results suggest that the mini-Flotac method is a valid, sensitive, and potentially low-cost alternative technique for use in the diagnosis of this yeast infection in birds.
Macrorhabdus ornithogaster (from the Greek words macrorhabdus for long rod and ornithogaster for stomach of bird) is an anamorphic Ascomycota yeast which can infect many species of birds (1). Its phylogenetic analysis and growth and metabolic characteristics were recently described (1-3). It colonizes the narrow junction (isthmus) of the glandular stomach (proventriculus) and grinding stomach (ventriculus) of birds and has not been identified elsewhere in the body or in the environment. The vegetative cells are elongated (2 to 20 m) and divided by fission. The cells are single or in short chains of two to four. Ascospores are not formed. It is Gram positive, but only the cytoplasm stains with Gram stain (2). In mucosal scrapings and in the feces of infected birds, the organism is a stiff, straight rod, 20 to 80 m long and 2 to 3 m wide, with rounded ends. In some circumstances, the long rods may bend slightly in a gentle curve. Viewed the organism is directly in a wet mount, small oblong refractile structures, the nuclei, found at regular intervals, are readily seen. Infection by M. ornithogaster has been described worldwide in a wide range of bird species, including chickens, turkeys, ostriches, several species of parrots, passerine species, and captive-bred and wild finches (3). There are different opinions on whether M. ornithogaster can cause disease. In fact, it was detected in its hosts both with and without obvious clinical signs. The majority of M. ornithogaster infections are benign or cause little detectable disease. Healthy birds can shed yeast cells while appearing normal on physical examination, whereas sick birds may not shed them continuously (4). The in vivo diagnosis of M. ornithogaster infection is based on the evaluation of clinical signs and/or on the microscopic examination of simple direct smears of fresh feces, possibly stained with a quick stain or Gram stain. M. ornithogaster cells in the feces can be also detected by PCR (3) and by culture methods using cloacal cotton-tipp...