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29 April 2019 An In Vitro Assay of Disinfectants on the Viability of Heterakis gallinarum Eggs
Katherine Lynn Cupo, Robert Byron Beckstead
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Nematodes are widespread and common in poultry. Disinfectants are used to reduce infection rates in poultry houses, but there is little documentation of their effectiveness. An in vitro assay was developed to test the efficacy of products to damage Heterakis gallinarum eggs, and nine disinfectants and chemicals commonly used in the poultry industry were tested. Embryonated eggs of H. gallinarum were pipetted into wells of plastic cell culture plates (250–300 eggs/well in water). Measured amounts of test articles were added to the suspensions for 2, 4, 6, or 24 hr. After exposure, eggs were washed with water and treated with trypan blue (1 ml of 0.4% solution, added to each well) at room temperature for 2 min. Eggshell integrity was determined microscopically by counting the number of eggs that were clear (intact) or that contained blue dye (compromised). As a test of embryo viability, five eggs per well from treatments containing compromised eggs were transferred to a Petri dish and hatched manually, using forceps to open the eggshell. Released larvae were then observed for signs of controlled movement. In a test of Clorox bleach (NaOCl), Green Klean, Decon7, Kem San, PLT, Virkon S, NaCl, dry limestone (CaCO3), and diesel fuel, only NaOCl (bleach) and Green Klean damaged the eggshell, and only 20,625 ppm of NaOCl rendered the larvae nonviable.

Katherine Lynn Cupo and Robert Byron Beckstead "An In Vitro Assay of Disinfectants on the Viability of Heterakis gallinarum Eggs," Avian Diseases 63(3), 511-513, (29 April 2019).
Received: 14 August 2018; Accepted: 20 March 2019; Published: 29 April 2019
embryonated eggs
Heterakis gallinarum
Histomonas meleagridis
in vitro assay
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