Fig. 1
From: The level of embryonation influences detection of Ostertagia ostertagi eggs by semi-quantitative PCR
Schematic overview of the study design. Anaerobic samples (n = 9 × 2) contained 10 g of freshly recovered faeces with Ostertagia ostertagi eggs and stored in vacuum bags under anaerobic conditions. Samples were subjected to either 4 °C or 25 °C for duration of 0 to 336 h. To stop egg development the samples were frozen at -20 °C. Subsequently, O. ostertagi were isolated and differentiated to developmental stage. Finally, 25× O. ostertagi eggs or L1 were counted and transferred to a clean petri dish. This was done in triplicates of each sample and ITS2 copies were quantified by qPCR. Aerobic samples (n = 9 × 2) were produced identically to anaerobic samples but eggs were isolated from faeces before storage and differentiated immediately at each time point. A total of 108 biological replicates were subjected to qPCR semi-quantification. Abbreviations: EPG, eggs per gram; ITS2, Internal Transcribed Spacer 2; qPCR, real-time semi-quantitative polymerase chain reaction