title: Quantification of Eimeria acervulina in faeces of broilers: comparison of McMaster oocyst counts from 24h faecal collections and single droppings to real-time PCR from cloacal swabs.
abstract: Coccidiosis is an economically important disease in chickens, caused by infection with Eimeria species parasites. Diagnosis of coccidiosis is frequently based on oocyst enumeration in pooled faecal samples or litter. In studies on infection dynamics and for monitoring in the field, samples from individual chickens may be more appropriate as these support the determination of infection status of individual birds and more accurately reflect oocyst output at time of sampling. Faecal samples from individual birds can be collected, but the counting procedure limits the number of samples that can be processed and unequivocal microscopic differentiation between Eimeria species is very difficult. A test that overcomes these drawbacks would improve efficiency and quality of the diagnosis. The aim of this study was to compare two methods for Eimeria oocyst quantification in samples from individual birds. A real-time PCR that quantifies oocysts in cloacal swabs (qPCR) and oocyst counts in single droppings were compared to the standard procedure of oocyst counts in bulked 24h faeces. Faecal samples were collected daily from 30 broiler chickens, inoculated with different doses of Eimeria acervulina. The three techniques produced comparable oocyst counts for all inoculation doses. Single dropping counts are applicable for small sample sizes and when a single Eimeria species is used. For larger sample sizes qPCR is preferable as it can be carried out on samples that have been frozen for storage. Furthermore, qPCR can identify and quantify different Eimeria species, which makes it a valuable diagnostic tool for field or experimental work.
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. f.c.velkers@uu.nl
authors.forename: F C
authors.initials: FC
authors.lastname: Velkers
authors.forename: D P
authors.initials: DP
authors.lastname: Blake
authors.forename: E A M
authors.initials: EA
authors.lastname: Graat
authors.forename: J C M
authors.initials: JC
authors.lastname: Vernooij
authors.forename: A
authors.initials: A
authors.lastname: Bouma
authors.forename: M C M
authors.initials: MC
authors.lastname: de Jong
authors.forename: J A
authors.initials: JA
authors.lastname: Stegeman
mesh_terms: D000818:Animals; D002645:Chickens; D002988:Cloaca / Q000469:parasitology; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D004539:Eimeria / Q000502:physiology*; D005243:Feces / Q000469:parasitology; D008297:Male; D009865:Oocytes / Q000502:physiology; D010270:Parasite Egg Count / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011201:Poultry Diseases / Q000469:parasitology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors