Scrapie is considered an endemic disease in both sheep and goats in Greece. However, contrary to sheep, in goats more than one prion protein (PrP) polymorphism has been recognized as a candidate for resistance breeding against the disease. For an impression, candidates which are circulating, (i) brain samples (n = 525) from scrapie-affected (n = 282) and non-affected (n = 243) animals within the national surveillance program, and (ii) individual blood samples (n = 1708) from affected (n = 241) and non-affected (n = 1467) herds, in a large part of mainland Greece and its islands, were collected and assayed. A dedicated Taqman method was used to test for amino acid polymorphisms 110T/P, 146N/S/D, 211R/Q, and 222Q/K. Highly prevalent genotypes were 110TT, 146NN, 211RR, and 222QQ. The frequencies of polymorphisms in blood and negative brain samples for codons 110P, 211Q, and 222K were 4.0%, 3.0%, and 1.9%, respectively, while 146D (0.7%) was present only on Karpathos island. Codon 110P was exclusively found in scrapie-negative brains, and homozygous 110P/P in two scrapie-negative goats. It is concluded that breeding programs in Karpathos could focus on codon 146D, while in other regions carriers of the 110P and 222K allele should be sought. Case-control and challenge studies are now necessary to elucidate the most efficient breeding strategies.
The aim of the present work was to analyze the antibiotic resistance of Pasteurella multocida isolates from sheep, goats, cattle and pigs originated from eight different Greek prefectures, to determine the class(es) of the tet genes, to investigate any possible association with plasmids or/and chromosomes and to study the geographical distribution of the resistant isolates. Eighteen out to one hundred isolates were resistant to tetracycline with a minimal inhibitory concentration (MIC) higher than 128 μg/ml. The tetH gene was found in thirteen (72.2%) and the tetB gene was found in four isolates (22.2%). After treatment with the restriction enzymes EcoRV, EcoRI, PstI and BspHI, the tetB gene showed a new polymorphism. Partial sequencing of the tetH and tetB genes revealed five polymorphisms in the tetB gene and two polymorphisms in the tetH gene; phylogenetic and molecular evolutionary analyses were also conducted. Both tetB and tetH genes revealed patterns clearly differentiated from the rest of Pasteurella species meaning that the genes could be transferred from a non Pasteurella species. The results presented provide further knowledge and evidence of the role played by commensals as of resistance determinants.
ABSTRACT. Scrapie is a slowly progressive infectious disease of sheep and goats that causes degeneration of the central nervous system. Scrapie is one of several transmissible spongiform encephalopathies (TSEs), like the bovine spongiform encephalopathy (BSE). In sheep, polymorphisms at codons 136, 154 and 171 of the host gene PRPN that encodes the PrP protein, are known to be closely linked to susceptibility or resistance to natural and experimental classical scrapie. In many countries, but not in Greece, breeding programs have been implemented to increase genetic resistance. This study was supported mainly by the private initiatives of farmers willing to improve their flocks by increasing the resistance to scrapie. Thus, the PrP genotypes (of the three mentioned codons) from 5815 blood samples of clinically healthy rams from 160 healthy flocks during the period 2012 -2016 were determined. Additionally, 1399 blood samples were genotyped only for the 171 codon. Samples were analyzed by Real Time PCR (TaqMan probes) with specific labeled probes. Our results showed an increased percentage of the two genotypes, ARR/ARR and ARR/ ARQ linked with resistance to the disease (27.29% and 34.6%, respectively) and relatively reduced percentage of the genotype ARQ/ARQ (24.23%) which is associated with susceptibility to disease and is the most common genotype in the Greek flocks. This joined effort has resulted in the establishment of an important number of farms with an increased population of genetically resistant rams to classical scrapie.
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