Batches of infection were carried out to ensure accuracy, repeatability and enough animals for each data point.
The cohort was then grouped using randomly selected animals from the batches of infection (Supplementary information, Table S3).
H&E-stained head sections were examined by a pathologist masked to groups and scored as one batch for signs of infection and inflammation.
A further issue is that production of each new vaccine batch requires infection of cattle followed by application of ticks, during which genetic recombination may occur, and finally the combination of material from ground ticks, each carrying a variable amount of one of the three vaccine stocks [ 8].
Validation of this micro-array experiment was performed by deep sequencing analysis of RNA samples derived of 4 batches of approximately 150 embryos at the 5 day time point of infection and 4 non-infected batches of embryos.
In order to map variation related to worm burden and the levels of cytokines that were produced in response to infection, rather than possible confounding covariates, we used General Linear Models (GLMs) to remove all covariance associated with differences in age, batch of eggs used for infection, and maternal genotype.
To calculate the viral titer to be used in the blood mixture of infection assays, different batches of Ae. albopictus STDEN1-F2 collected in La Réunion in 2006 were infected with different viral titers: 105, 106, 107, 108, and 109 pfu/mL and dissemination rates were estimated.
The average age of infection across all batches was 43 days of age.
Therefore, it is not surprising that the amino acids quantification did not demonstrate any limitations at the time of infection for either the batch or the perfusion cultures.
One month after the last immunization, all lambs were infected intravenously with 2 ml of the homologous viable batch of A. phagocytophilum with an approximate infection dose equal to 0.5 × 10 infected neutrophils per ml.
