Automated enumeration of Eimeria oocysts in feces for rapid coccidiosis monitoring

Coccidiosis represents a major driver in the economic performance of poultry operations, as coc-cidia control is expensive, and infections can result in increased feed conversion ratios, uneven growth rates, increased co-morbidities with pathogens such as Salmo-nella, and mortality within flocks. Shifts in broiler produc-tion to antibiotic-free strategies, increased attention on pre-harvest food safety, and growing incidence of anti-coc-cidial drug resistance has created a need for increased understanding of interventional efficacy and methods of coccidia control. Conventional methods to quantify coc-cidia oocysts in fecal samples involve manual microscopy processes that are time and labor intensive and subject to operator error, limiting their use as a diagnostic and mon-itoring tool in animal parasite control. To address the need for a high-throughput, robust, and reliable method to enumerate coccidia oocysts from poultry fecal samples, a novel diagnostic tool was developed. Utilizing the PIPER instrument and MagDrive technology, the diag-nostic eliminates the requirement for extensive training and manual counting which currently limits the application of conventional microscopic methods of oocysts per gram (OPG) measurement. Automated microscopy to identify and count oocysts and report OPG simplifies analysis and removes potential sources of operator error. Morphometric analysis on identified oocysts allows for the oocyst counts to be separated into 3 size categories, which were shown to discriminate the 3 most common Eimeria species in commercial broilers, E. acervulina, E. tenella, and E. maxima. For 75% of the samples tested, the counts obtained by the PIPER and hemocytometer meth-ods were within 2-fold of each other. Additionally, the PIPER method showed less variability than the hemocy-tometer counting method when OPG levels were below 100,000. By automated identification and counting of oocysts from 12 individual fecal samples in less than one hour, this tool could enable routine, noninvasive diagnos-tic monitoring of coccidia in poultry operations. This approach can generate large, uniform, and accurate data sets that create new opportunities for understanding the epidemiology and economics of coccidia infections and interventional efficacy.

Automated summary

Coccidiosis is a significant factor affecting poultry operations, leading to increased costs and negative impacts on growth and mortality. Conventional methods of diagnosing and monitoring the disease are time-consuming, labor-intensive, and prone to errors. A novel diagnostic tool, utilizing automated microscopy and morphometric analysis, has been developed to accurately and efficiently quantify coccidia oocysts from poultry fecal samples. This tool can provide valuable insights into the epidemiology and economics of coccidiosis, improving intervention strategies.