Peripheral neuropathy caused by fenugreek (Trigonella foenum-graecum) straw intoxication in cattle and experimental reproduction in sheep and goats.

Trigonella foenum-graecum (fenugreek) is a legume widely used as a food supplement in humans and less frequently in ruminants. Toxicity has been described sporadically in ruminants grazing mature fenugreek plants or stubble; however, the pathological features are unclear. This report describes a natural outbreak of intoxication in cattle fed fenugreek straw and the experimental reproduction using 8 sheep and 8 goats. Affected cattle presented clinical signs approximately 1 month after consuming the straw and 100 of 400 cattle (25%) were affected, of which 60 of 100 (60%) died or were euthanized. Clinical signs were characterized by proprioceptive positioning defects with abnormal postures and weakness of hindlimbs. Forelimbs were also affected in severely affected animals, and cattle became recumbent. Locomotion was characterized by trembling, and some cattle showed high-stepping movements of their forelimbs and knuckled over in their fetlocks. Experimental intoxication induced clinical signs only in sheep and were similar to cattle, although with signs starting in the forelegs. Gross and microscopic lesions were similar in spontaneous and experimental intoxications. Macroscopic changes corresponded with muscular hemorrhages and edema, mainly surrounding the peripheral nerves. Microscopic examination only demonstrated lesions in the distal peripheral nerves, which included edema, hemorrhages, and Wallerian degeneration. Neurofilament immunohistochemistry revealed altered axon labeling and S100 showed a decrease in myelin intensity and loss of its typical compact arrangement around axons. Biochemical and hematological abnormalities included elevated levels of muscle and liver enzymes and thrombocytopenia. These findings indicate that fenugreek straw induces peripheral neuropathy in cattle and sheep, but not in goats.

Low sequence diversity of the prion protein gene (PRNP) in wild deer and goat species from Spain.

The first European cases of chronic wasting disease (CWD) in free-ranging reindeer and wild elk were confirmed in Norway in 2016 highlighting the urgent need to understand transmissible spongiform encephalopathies (TSEs) in the context of European deer species and the many individual populations throughout the European continent. The genetics of the prion protein gene (PRNP) are crucial in determining the relative susceptibility to TSEs. To establish PRNP gene sequence diversity for free-ranging ruminants in the Northeast of Spain, the open reading frame was sequenced in over 350 samples from five species: Iberian red deer (Cervus elaphus hispanicus), roe deer (Capreolus capreolus), fallow deer (Dama dama), Iberian wild goat (Capra pyrenaica hispanica) and Pyrenean chamois (Rupicapra p. pyrenaica). Three single nucleotide polymorphisms (SNPs) were found in red deer: a silent mutation at codon 136, and amino acid changes T98A and Q226E. Pyrenean chamois revealed a silent SNP at codon 38 and an allele with a single octapeptide-repeat deletion. No polymorphisms were found in roe deer, fallow deer and Iberian wild goat. This apparently low variability of the PRNP coding region sequences of four major species in Spain resembles previous findings for wild mammals, but implies that larger surveys will be necessary to find novel, low frequency PRNP gene alleles that may be utilized in CWD risk control.

Polimorfismo do gene da proteína prion celular (PrPC) e imunohistoquímica de tecido linfóide em ovinos / Polymorphism of cellular prion protein (PrPC) and immunohistochemistry of lymphoid tissue of sheep

O objetivo deste trabalho foi avaliar o polimorfismo do gene da proteína prion celular (PRPN) de ovinos introduzidos numa propriedade onde ocorreu um surto de scrapie, e relacionar com a suscetibilidade à doença por meio da análise da presença da proteína prion celular alterada (PrPSc), utilizando imunohistoquímica (IHQ) de tecido linfóide associado à mucosa reto-anal. Foram avaliados 42 ovinos, mestiços Texel. Eram fêmeas entre um e oito anos de idade, sendo que sete (16,67%) ovelhas foram introduzidas adultas na propriedade em 2006. As demais, 83,33%, eram nascidas na fazenda. A genotipagem do PRPN foi feita pela análise do polimorfismo de comprimento de fragmento de restrição - RFLP ("Restriction Fragment Lenght Polimorphism"). O genótipo ARQ/ARQ foi o mais freqüente, encontrado em 73,81% dos animais, seguido do genótipo ARR/ARQ, com 16,67% e do ARQ/VRQ, com 9,52%. Os alelos ARH e AHQ não foram encontrados nestes animais. O resultado da IHQ foi negativo em todas as amostras. Não foi possível, portanto, estabelecer uma relação entre genótipo e maior susceptibilidade ao scrapie, devido à ausência de PrPSc na amostras examinadas. No Brasil, há poucos dados de genotipagem do gene da proteína prion celular (PRNP) em ovinos e, até o momento, nenhum tipo de controle baseado em cruzamentos direcionados foi implementado.

The aim of this work was to study the polymorphism of the prion protein gene (PRNP) of a sheep flock raised in a farm where a scrapie outbreak had occurred, and to relate to disease susceptibility of possible animals infected with altered prion protein (PrPSc), by immunohistochemical analysis of recto-anal mucosa-associated lymphoid tissue (RAMALT). Forty two sheep, crossbred with Texel, Ile de France, Dorper and Suffolk were used. Females were between one and eight years old, and seven (16.67%) were adult ewes when they entered the flock in 2006. The rest, 83.33% were born in the farm. The PRNP genotyping was performed by RFLP ("restriction fragment length polymorphism") analysis. The most frequent genotype was ARQ/ARQ, found in 73.81% of the animals, followed by ARR/ARQ, with 16.67% and ARQ/VRQ, with 9.52%. The ARH and AHQ alleles were not found. All RAMALT samples were negative in immunohistochemical analysis. It was not possible to establish a relation between PRNP polymorphisms and susceptibility to scrapie, due to the lack of positive samples to PrPSc. In Brazil, there is little available PRNP genotyping data of sheep and, so far, no type of controlled breeding scheme for scrapie has been implemented.