Effect of different floatation solutions on E. tenella oocyst purification and optimization of centrifugation conditions for improved recovery of oocysts and sporocysts

• Published in: Experimental parasitology Vol. 217; p. 107965
• Main Authors: Qi, Nanshan, Liao, Shenquan, Abuzeid, Asmaa M.I., Li, Juan, Wu, Caiyan, Lv, Minna, Lin, Xuhui, Hu, Junjing, Xiao, Wenwan, Sun, Mingfei, Li, Guoqing
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-10-2020
• Subjects: Eimeria tenella; Oocysts; Purification; Sodium hypochlorite; Sporocysts; Eimeria tenella; Sodium hypochlorite; Purification; Sporocysts; Oocysts
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2020.107965

Saturated salt floatation method is widely used for coccidian oocyst purification. However, the repeated procedures and inefficient oocysts recovery rate are a continuous challenge. This study aimed to investigate the best suitable floatation solution, along with optimal centrifugation speed and time for Eimeria tenella (E. tenella) oocyst and sporocyst purification. Different floatation solutions i-e, saturated salt, Sheather's sugar and sodium hypochlorite (NaClO) at 20–60% concentrations were used to purify oocyst. It was found that about 96.99% oocysts (8609×g for 10 min) were recovered under these conditions without any effect on the viability of sporocysts. The recovery rate of oocysts using 50% NaClO (V/V) was significantly higher than 35% saturated salt flotation solution (P < 0.05). The optimal method for purification of oocysts based our experimentation was centrifugation at 8609×g for 3 min using 50% NaClO floatation solution, and the optimized centrifugation conditions for improved recovery of sporocysts (about 99.3%) were at 2152×g for 5 min. The present study provided a better method for the coccidian oocyst purification, which could be successfully adopted as a better alternative to existing techniques commonly used for investigations/research pertaining to coccidia.
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•We investigated the effect of centrifugation speed on E. tenella oocyst and sporocyst recovery.•We investigated the effect of centrifugation time on E. tenella oocyst and sporocyst recovery.•We determined the improved methods for the coccidian oocyst purification.•We determined the speed and time of centrifugation did not affect the viability of sporocysts.