Diego José Zanzarini Delfiol

Mycobacterium DNA detection in liver and skin of a horse with generalized sarcoidosis

Sarcoidosis is a rare equine skin disease characterized primarily by an exfoliative and granulomatous dermatitis but also presenting granulomatous inflammation of multiple systems. The current report presents the clinical and histopathological findings of sarcoidosis in a 16-year-old American Quarter Horse gelding with nested polymerase chain reaction Mycobacterium spp. DNA detection within hepatic and skin samples. Mycobacterium spp. may play a role in the pathogenesis of equine sarcoidosis as has been proposed for human sarcoidosis.

Equine poisoning by coffee husk (Coffea arabica L.).

BackgroundIn Brazil, coffee (Coffea arabica) husks are reused in several ways due to their abundance, including as stall bedding. However, field veterinarians have reported that horses become intoxicated after ingesting the coffee husks that are used as bedding. The objective of this study was to evaluate whether coffee husk consumption causes intoxication in horses.ResultsSix horses fed coast cross hay ad libitum were given access to coffee husks and excitability, restlessness, involuntary muscle tremors, chewing movements and constant tremors of the lips and tongue, excessive sweating and increased respiration and heart rates were the most evident clinical signs. Caffeine levels were measured in the plasma and urine of these horses on two occasions: immediately before the coffee husks were made available to the animals (T0) and at the time of the clinical presentation of intoxication, 56 h after the animals started to consume the husks (T56). The concentrations of caffeine in the plasma (p < 0.001) and urine (p < 0.001) of these animals were significantly greater at T56 than at T0.ConclusionsIt was concluded that consumption of coffee husks was toxic to horses due to the high levels of caffeine present in their composition. Therefore, coffee husks pose a risk when used as bedding or as feed for horses.

Prevalence of the Glycogen Branching Enzyme Deficiency Mutation in Quarter Horses in Brazil

ABSTRACT Glycogen branching enzyme deficiency (GBED) is a fatal autosomal inherited disease affecting horses and caused by a nonsense mutation (c.102C>A) in the GBE1 gene. This disease strongly impairs the glycogen metabolism of affected animals resulting in abortion or early death of foals. Although understanding the prevalence of heterozygous for GBED is imperative to estimate the impact of the disease, prevalence studies have yet to be performed in Brazil. The aim of this study was to determine the prevalence of heterozygous of the GBED mutation in a population of Quarter Horses in Brazil. Blood samples were obtained from 742 animals competing in distinct disciplines including cutting, halter, race, reining, and barrel racing. All samples were submitted to DNA purification, amplification, and sequencing of the target fragment. The overall GBED prevalence of carriers was 7.95% (59/742). The prevalence of heterozygotes by discipline was considered higher in cutting (19.75% [32/162]) and reining (10% [16/160]) subgroups followed by barrel racing (5% [8/160]) and halter (3% [3/100]) horses. There were none heterozygous in the racing subgroup. These results demonstrate the GBED causative mutation in the Brazilian Quarter Horse herd and suggest the occurrence of the disease. Therefore, GBED is a disease that should be considered in the differential diagnosis of abortion and stillbirths, especially if suspect foals are products of two heterozygous. Besides testing aborted foals for the GBED mutation, the molecular diagnosis should be recommended to owners and breeders, at least for those breeding the at‐most‐risk subgroups of horses, to guiding their mating selections and, therefore, prevent the transmission of the mutation. HighlightsGlycogen branching enzyme deficiency (GBED) is a fatal autosomal recessive monogenic disease, causing abortions and stillbirths.The prevalence of GBED mutation carriers was unknown among Quarter Horses in Brazil.The prevalence of heterozygous horses for the GBED mutation in Brazil is 7.94%.

Influência da suplementação nutricional sobre o metabolismo energético e enzimas musculares em cavalos de Team Penning

O Team Penning (TP) e uma modalidade equestre que exige velocidade maxima. Nesse tipo de exercicio ocorre o rapido consumo de energia pela via anaerobica. Diversos suplementos tem sido desenvolvidos para melhorar o desempenho e diminuir lesoes musculares em equinos atletas. Objetivou-se neste estudo avaliar a influencia da suplementacao com Tonnus JCR® nos parâmetros fisiologicos, metabolismo energetico e muscular no treinamento de TP. Foram utilizados 12 equinos da raca Quarto de Milha, atletas de TP, separados em dois grupos, suplementado (GS) e nao suplementado (GNS), avaliados em dois treinos. Foi realizado o exame fisico, avaliacao das concentracoes plasmaticas de glicose e lactato, e concentracoes sericas de creatinoquinase (CK), aspartato aminotransferase (AST) e lactato desidrogenase (LDH). As amostras de sangue foram colhidas durante e apos os treinos em 12 diferentes momentos (M0 - M11). A suplementacao foi realizada no intervalo entre os treinos. No segundo treino, verificou-se diminuicao nas concentracoes de glicose no GS quando comparado ao treino 1 (p = 0,01); no GNS nao houve diferenca entre os treinos (p = 0,13). Nao houve diferenca nas concentracoes de lactato entre grupos e treinos. Os valores da concentracao de CK no treino 1 nao apresentaram diferenca entre os grupos (p = 0,83) e momentos (p = 0,16). Quando comparadas as concentracoes de CK entre os grupos no treino 2, observaram-se concentracoes menores no GS (p = 0,02). Ao se comparar o GS antes e apos a suplementacao, as concentracoes de CK foram menores no treino 2 (p = 0,01). No GNS nao houve diferenca entre os treinos (p = 0,51). As enzimas AST e LDH nao diferiram entre os grupos nos treinos 1 e 2. Concluiu-se que a suplementacao influenciou de forma positiva, diminuindo as concentracoes de glicose e os niveis de CK, o que indica menor injuria muscular pos-treino.

Validation of high-resolution melting analysis as a diagnostic tool for endothelin receptor B mutation in American Paint horses and allele frequency estimation

Overo lethal white foal syndrome (OLWFS) is a genetic disorder caused by a dinucleotide mutation in the endothelin receptor type B (EDNRB) gene leading to the death of affected foals shortly after birth. The use of rapid and reliable genetic testing is imperative for the early diagnosis of the mutation avoiding, therefore, either additional suffering or the production of affected animals. In the present study, we developed and validated a high-resolution melting (HRM) genotyping assay to detect the OLWFS causative mutation, and we also determined the frequency of heterozygotes among American Paint horses in Brazil. The HRM genotyping assay resulted in a high sensitivity, specificity, and positive and negative predictive values. The overall estimated frequency of heterozygotes was 21.6%; however, this frequency increased to 89.5% when considering only overo horses. The HRM assay optimized here was a reliable and suitable method for the detection of the dinucleotide mutation observed in the EDNRB gene resulting in a fast, accurate, and precise diagnostic tool. The causative gene mutation of OLWFS is present in heterozygosity in the American Paint Horse population in Brazil and is highly frequent among overo horses.

Estimation of the Allele Frequency of Type 1 Polysaccharide Storage Myopathy and Malignant Hyperthermia in Quarter Horses in Brazil

Abstract Type 1 polysaccharide storage myopathy (PSSM1) and malignant hyperthermia (MH) are autosomal dominant genetic diseases caused by a point mutation in equine GYS1 and RYR1, respectively. Although the prevalence of PSSM1 and MH have been described in other countries as causes of myopathy in horses, there are no studies evaluating the allele frequency of those mutations in Brazil. The aim of the present study was to determine the allele frequency of the mutated alleles causing susceptibility for PSSM1 and MH mutations in Quarter Horses (QHs) in Brazil. Deoxyribonucleic acid was purified from blood samples of 741 QHs used in five competitive disciplines (i.e., barrel racing, cutting, halter, racing, and reining). The PCR was optimized to identify the mutations using primers designed from the sequence of equine GYS1 and RYR1. The allele frequencies and the prevalence of PSSM1 were 0.034 and 6.7%, respectively. The highest allele frequency for the GYS1 mutated gene was identified in halter horses (0.163), and the gene mutation was not observed in racing animals. The mutation in the RYR1 was not identified in any of the sampled horses. The PSSM1 allele frequency observed in the present study indicates the relevance of the disease in the differential diagnosis of myopathies in horses. In addition, the absence of the RYR1 causative mutation suggests little importance of MH as a genetic muscle disorder in the Brazilian QHs. HighlightsThe allele frequencies and the prevalence of type 1 polysaccharide storage myopathy (PSSM1) were 0.034 and 6.7%, respectively.The highest allele frequency for the GYS1 mutated gene was identified in halter horses (0.163).In 48.8% (20/41) of the stud farms where the samples were obtained, at least one animal was positive for PSSM1.The mutation in the RYR1 was not identified in any of the sampled horses.

USO DE CISPLATINA INTRALESIONAL PARA O TRATAMENTO DE CARCINOMA DE CÉLULAS ESCAMOSAS EM OVINO DA RAÇA SANTA INÊS

The squamous cell carcinoma (SCC) is a malignant tumor of epidermal cells, the keratinocytes. It is a slow growing cancer, locally invasive and occasionally metastatic. Is directly associated with prolonged exposure to ultraviolet light, in glabrous areas or at sites lacking skin pigment. The tumors can have productive or erosive appearance and is diagnosed histologically. Treatment depends on the tumor extent, location and expected the animal function. Cisplatin is the most effective anticancer agent in the treatment of solid tumors, however its systemic use has been limited due to toxicity. The intratumoral administration allows for maximum concentration of the drug while minimizing effects of exposure to other tissues. We herein a SCC tumor in a sheep, Santa Ines breed, near the coronary border of the lateral digit of the left pelvic limb. The diagnosis of SCC was determined by clinical examination followed by histopathological evaluation of the mass. The animal was treated with intralesional cisplatin (Fauldcispla®, Libbs) in an aqueous vehicle at a dose of 0.97 mg / cm3 of tumor, injected in parallel planes, every two weeks, and four applications. The antineoplastic therapy was satisfactory as shown by the complete regression of the tumor mass, lack of adverse effects and no recurrence.