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Additional file 1. Principle of the three ELISA tests to detect bovine PrPSc and PrPres.

Additional file 2. PrPSc digestion by keratinase of bovine brain after heat treatment at 115 °C. BSE infected brain in lanes 4, 7 and 10-13, negative control brain in lanes 2 and 3. Material in lanes 2 and 4 was heated at 115 °C in presence of detergent before digestion. Lanes 3 and 10-13: non-heated material digested by KE. In the heated BSE sample in lane 4 no PrP-specific immunoreactivity has remained neither throughout the lane nor in the high molecular mass region at the top, while in the non-heated material there was (lane 10). Lanes 1 and 9, mixture of molecular mass markers SeeBlue and MagicMark XP for which migration positions are indicated in kDa at the left; lanes 5, 6 and 8, no sample applied. Top and running front are indicated with arrow heads and arrows, respectively. Antibody used: 94B4 (0.2 μg mL-1). Gel was run in MOPS buffer. Symbols: TE = tissue equivalents; MM = molecular mass markers; KE = keratinase; + = treatment applied.

Additional file 4. Sample ranking to their relative levels of N-terminal PrPres epitopes of antibodies P4 and 12B2. Figure in histogram form to compare the ranking of goat study sample series from high to low N-terminal epitope in ISS-WB and Triplex-WB.

Additional file 6. Graphic comparison of the PrPres double and single triplet state in resp. CH1641 and scrapie/BSE. Figure showing difference in migration of PrPres bands in CH1641 scrapie samples, goat study sample UK-B2 and reference TSEs as obtained with mAbs Sha31 and SAF84 in Triplex-WB.

Additional file 2. Overview of the application of seven different biochemical analyses in goat and control samples. Table containing goat sample sets used in the specific type of analyses performed.

Additional file 5. Correlation between N-terminus data and molecular mass of the PrPres non-glycosylated band obtained by Triplex-WB. Figure in dotplot form showing the correspondence between total PrPres N-terminus level and molecular mass of non-glycosylated PrPres fraction.

Additional file 3. ISS-WB of goat TSE brain samples from different geographical regions with antibodies P4 and SAF84. Figure of ISS-WB with mAbs P4 and SAF84 on the set of goat study samples.

International audience; Contrasting the knowledge about prion diseases or TSEs in sheep, only a very limited number of strain typing studies are available in goats. Two cases deriving from the zoonotic bovine BSE epidemic were however detected in goats. During 2004–2012, over 70 TSE goat brain samples were collected from seven European countries and evaluated for TSE type/strain variation. A selection of these materials was chosen for in-depth analysis based on various criteria: tissue quality, genotype, broad geographical distribution, potential type variation. Of these, 37 cases were biochemically analysed (present study) and a subset of these subjected to bioassays in seven rodent models (see parallel study of Nonno et al.). Analyses of these isolates showed that in goats different PrPSc types exist, comparable to those found in sheep such as classical scrapie, CH1641-like scrapie and Nor98/atypical scrapie. However, classical scrapie isolates could be further differentiated in two molecular subtypes based on the PK susceptibility of the N-terminus and on the molecular weight of PrPres. Importantly, the two subtypes of classical scrapie showed different geographical distribution, as one subtype was only detected in goats deriving from Italy and France. These analyses also show, that none of the field samples exhibited BSE-like features and offer a comprehensive set of robust molecular PrPres properties to exclude suspicion for the presence of BSE. These are: molecular mass of the triplet bands, presence of N-terminus epitope, glycoprofile markers, a dual population marker and the structural stability of PrP-core.This work was made possible by national support and by the following European grants: Neuroprion (FP6-FOOD-506,579), GoatBSE (FOOD-CT-2006–36,353) and GOAT-TSE-FREE (EMIDA-ERA NET). We want to memorize Jorg G Jacobs, who died in 2017. Jorg developed and performed the Triplex-WB system as well as characterized antibodies like 94B4, 12B2, 9A2, 6C2, SAF84 and L42. He also was crucial in the distinction of C-, L- and H-type BSE and in 2011 he showed elegantly the allotype composition of prion material in heterozygous sheep by the use of Endo-LysC.

Additional file 1: Summary of original experiment. This document gives an overview of the inoculations carried out in cattle and wild-type mice with references to the animal numbers in the original, published study [2].

Prion diseases are a group of animal and human neurodegenerative, albeit transmissible diseases. The lack of effective preventive and therapeutic approaches represents a serious problem in their management. This is especially true for those prion diseases of animals which behave like infectious and contagious diseases. In the case of sheep scrapie, the well- known role of variations in the prion protein gene (PRNP) in conferring resistance/susceptibility represents an opportunity which has been exploited to select populations genetically resistant to the disease. The recent description of Camel prion disease (CPrD) in Algeria and Tunisia and the suspicion falling into the category of infectious prion diseases, make urgent to investigate the possible existence of genetic determinants useful for its control. Herein, we investigated PRNP variability in 232 animals from six dromedary populations (Azawad, Hybrid, Naili, Rguibi, Sahraoui, Targui) in Algeria. A Gly69Ser mutation was observed in a single animal of the Targui population and a Gly134Glu polymorphism in the Azawad, Hybrid and Rguibi populations, with a frequency of the 134Glu allele of 2.6%, 7.7% and 7.1%, respectively. Although our work highlights a low variability of Algerian dromedary PRNP, as a possible indication of a recent evolutionary history of CPrD, it offers also evidence of PRNP variants whose role in prion disease resistance/susceptibility deserve to be deepened.