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Objective To establish a polymerase chain reaction coupled with restriction fragment length polymorphism (PCR-RFLP) method which can identify deer blood. Methods We analyzed DNA sequences of cytochrome b (cytb) from different species (i.e. sika deer, red deer, pig, ox, sheep, and duck) and selected two DNA restriction enzymes of EcoRV and MseI to distinguish deer from other species and to sika deer from red deer, respectively. Genomic DNA of every sample was extracted by blood genomic DNA extraction kit. After PCR of cytb fragment, digestion by EcoRV or MseI was conducted and the results were analyzed. The capacity of this method to identify different proportional blood mix from different species also was determined. Results Using the PCR-RFLP method, EcoRV digestion made two fragments of 287 bp and 185 bp in deer, but no digestion in other species; MesI digestion made two fragments of 281 bp and 191 bp in sika deer, but three fragments of 281 bp, 126 bp, and 65 bp in red deer. The 191 bp fragment was a characteristic marker of sika deer, and the 126 bp was for red deer. The minimum detected proportion added to blood of deer with other sample was 3%, 6% of sika deer with red deer. Conclusion A PCR-RFLP method according to the sequences of cytb gene is established. This method can identify the blood of sika deer rapidly and conveniently.
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Worldwinde, cervids are considered an important source of infection and dissemination of a wide variety of pathogens, both for farm animals and humans. Among this diseases is sarcosporidiosis, which is a parasitic disease caused by Sarcocystis spp. (Protozoa: Apicomplexa). Most frequent clinical signs are hemolytic anemia, weakness, weight loss and decrease of growth and some species of Sarcocystis might cause abortions. The clinical disease in ruminants is fairly rare but the infection is very frequent. Infections are accumulative and the parasite does not generate immunity in any of the hosts. Ovine sarcosporidiosis is a serious issue in the some regions of Chile due to the macrocysts located in the muscle which means condemnation of the whole carcass. Sarcocystis spp. has been widely reported in red deer and other cervid species but in Chile the situation remains unknown. Nowadays there is little to no evidence of Sarcocystis in foreign deer in Chile and there is only one report of the parasite on pudu. The main goal of this study is to demonstrate the presence of Sarcocystis spp. in myocardium of red deer and fallow deer in Chile, and confirm the presence of Sarcocystis spp. in pudu. All cervid cases from 1994 to 2013 of the Institute of Animal Pathology of the Universidad Austral de Chile were reviewed. The animals selected were those in which a myocardium sample was taken. From the histopathological samples observed, it was found that 5 of the 9 red deer, 1 of the 4 fallow deer and in 11 of the 23 pudu there were Sarcocystis cysts in the myocardium. This study represents the first record for Chile of Sarcocystis spp. in myocardium of red deer and fallow deer. Stablishing the red deer, fallow deer and pudu as hosts of Sarcocystis aids to have a better understanding of the parasite epidemiology in Chile and the role of wild and captive cervids in the maintenance and spread of these parasites.(AU)
No mundo, os cervos são considerados uma fonte de infecção e propagação de uma grande variedade de patógenos para animais de criação e para os seres humanos. Entre estas doenças está a sarcosporidiosis, que é uma doença parasitária causada por Sarcocystis spp. (Protozoa: Apicomplexa). Os sinais clínicos mais comuns são anemia hemolítica, fraqueza, perda de peso e diminuição do crescimento e em algumas espécies de Sarcocystis podem causar abortos. A doença clínica em ruminantes é bastante rara, mas a infecção é muito comum. As infecções são cumulativos e o parasita não gera imunidade em nenhum dos seus hospedeiros. A Sarcosporidiosis ovina é um problema grave em algumas regiões do Chile devido a microcistos localizados no músculo provocando a reprovação total da carcaça. Sarcocystis spp. tem sido amplamente relatado em cervos vermelhos e outras espécies de cervídeos, mas no Chile a sua situação permanece desconhecida. Atualmente há pouca ou nenhuma evidência de Sarcocystis em cervos introduzidos no Chile e há apenas um relatório do parasita em pudú. O principal objetivo deste estudo é demonstrar a presença de Sarcocystis spp. no miocárdio no veado vermelho e cervo gamo no Chile e confirmar a presença de Sarcocystis spp. em pudus. Revisaram-se todos os casos de cervos desde 1994-2013 do Instituto de Patologia Animal da Universidad Austral de Chile. Os animais selecionados para o estudo foram aqueles em que se tomou amostra de miocárdio. Das amostras histopatológicas observadas, verificou-se que em cinco dos nove cervos vermelhos, em um dos quatro veados gamo e 11 dos 23 pudus tinham cistos de Sarcocystis no miocárdio. Este estudo representa o primeiro relatório para o Chile de Sarcocystis spp. no miocárdio de veados vermelhos e cervo gamo. Definir o veado vermelho, o cervo gamo e os pudú como anfitriões de Sarcocystis ajuda a uma melhor compreensão da epidemiologia deste parasita no Chile e o papel de cervos selvagens e em cativeiro para a manutenção e divulgação deste parasita.(AU)
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Animales , Ciervos/parasitología , Sarcocystis/parasitología , Sarcocistosis/epidemiología , ChileRESUMEN
Paratuberculosis is a disease caused by Mycobacterium avium subsp. paratuberculosis (MAP) that affects domestic and wild ruminants. The most common gross lesions are emaciation and corrugation and thickening of the mucosa of the small intestine. Mesenteric lymph nodes might be enlarged. For the present study, 14 red deer and 9 fallow deer from game reserves or venison farms were analyzed. The lesions found correspond to those found by other authors in other geographic locations, except for some differences in histopathological examinations. Among these differences, stands out that intestinal lesions were concentrated mostly in the ileum and granulomas were shown to be more frequent in this section of the intestine than in the corresponding lymph node. Furthermore, in multibacillary lesions the inflammatory infiltrate in the lymph nodes was mainly composed of macrophages. These differences may be due to individual variations of the animals, the stage of disease or a different strain of the pathogen. This study allowed to obtain basic information about the disease and to describe patterns of lesions found in red deer and fallow deer with prediagnosis of clinical paratuberculosis which were not described in the literature before.(AU)
Paratuberculosis é uma doença causada por Mycobacterium avium subsp. paratuberculosis (MAP) que afecta ruminantes selvagens e domésticos. As lesões macroscópicas mais comuns são ondulação e espessamento da mucosa do intestino delgado. Os linfonodos mesentéricos podem aparecer com volume aumentado. Para este estudo, foram analisados 14 veados vermelhos e 9 veados gamo de reservas de caça e fazendas de carne. As lesões encontradas correspondem à encontrada por outros autores em outras localizações geográficas, com exceção de algumas diferenças no exame histopatológico. Entre essas diferenças, sobressai que as lesões intestinais se concentraram principalmente no íleo, os granulomas ocorreram com maior frequência nesta seção do intestino que no seu correspondente linfonodo. Além disso, nas lesões bacterianas, o infiltrado inflamatório linfonodos linfáticos era composta principalmente por macrófagos. Estas diferenças podem ser devidas a variações individuais dos animais, o estádio da doença ou de uma estirpe diferente do agente patogénico. Este estudo permitiu obter informação básica sobre a doença e descrever padrões de lesões encontradas em veados e em gamos com pré-diagnóstico, de paratuberculosis clínica nunca antes descritas na literatura.(AU)
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Animales , Paratuberculosis/diagnóstico , Ciervos/microbiología , Mycobacterium avium/aislamiento & purificación , ChileRESUMEN
For rapid identification of Cervus nippon, C. elaphus and their hybridize samples, the specific PCR for mutual authentication of them was established based on the SNPs in COI and SRY sequence. C. nippon, C. elaphus and their hybridize samples were collected from different origins, total DNA of 24 identified samples were extracted, and the COI and SRY gene was seqenced. SNPs in the COI and SRY sequences of the samples were found by Clustul X 2.1 program. Primers for identifying C. nippon and C. elaphus were designed according to the SNP site, two multi-PCR reaction system were established to identify them. In addition, 24 samples which were randomly collected in different herbal medicine market were identified. The band special for C. nippon (232 bp)and band special for C. elaphus (518 bp) based on COI sequence,and the band special for C. nippon (803 bp)and band special for C. elaphus (425 bp) based on SRY sequence, were found using multi-PCR reaction, and three of the twenty-four samples were identified as the hybridize samples. The multi-PCR reaction system could be used to identify C. nippon, C. elaphus and their hybridize samples.
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OBJECTIVE: To determine the contents of antler peptides in Cervus elaphus yarkandensis from different locations and harvest periods to evaluate their qualities. METHODS: The concentration of antler peptides of Cervus elaphus yarkandensis was measured by BCA protein assay kit. The protein contents between differen regions and harvest time were compared. RESULTS: A good linear relationship was achieved for the calibration curve of antler peptides in the range of 20-2 000 μg · mL-1 (r=0.997 9). The average recovery was 99.06% with RSD of 2.08% (n=6). The contents of antler peptides in Cervus elaphus Linnaeus from different locations were significantly different. There were also differences in the antler peptides content of the same origin samples collected in different periods. CONCLUSION: The established method is simple, accurate and reproducible, which can be adopted for determination of antler peptides in Cervus elaphus Linnaeus.
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Objective To evaluate the effect of antler polypeptides from Cervus elaphus yarkandensis on MC3T3-E1 cells. Methods The concentration of antler polypeptides of Cervus elaphus yarkandensis was measured by BCA protein assay kit. MC3T3-E1 cells were cultured and analyzed by BCIP/NBT chromogenic alkaline phosphatase (ALP) staining kit. After being induced to form mineralized knot, the cells were stained by using Alizarin red staining. Three different concentrations (10, 1.0, 0.1 μg/mL) of antler polypeptides were analyzed by MTT method and micronutrients enzymes standard method to determine the effect of cell proliferation and ALP synthesis. Results The concentration of antler polypeptides was 0.07 mg/mL. The results of in vitro cell activity analysis showed that the positive rate of ALP was 90%and the mineralization knot was stained red. Compared with the control group, the different concentrations of antler polypeptides all showed the function of cell proliferation and the effect was dose-dependent after 3 d and 7 d. Compared with the control group, at the 3 d, three groups of antler polypeptides promoted synthesis and secretion of ALP (P<0.01) and the results showed a dose-dependent effect. Conclusion Antler polypeptides could obviously promote the proliferation of MC3T3-E1 cells and secretion of ALP, which indicated that antler polypeptides have certain effect on osteoporosis.
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The objective of this study was to estimate the direct and maternal genetic variance components for some growth traits in a red deer herd (Cervus elaphus scoticus) located in Queretaro, Mexico. Information between 1994 and 2003, consisting of 417 records of birth weight (BW), 169 weaning weight (WW), 168 six months weight (6MW) and 172 yearly weight (YW) was analyzed, which included the identification of 554 animals with 11 stags and 107 hinds. The fixed effects considered were: sex, year of birth and pregnancy number (P < 0.001). Three mixed models were used. Model 1 included the fixed effects and the direct additive genetic effect; Model 2 included those in 1 plus the maternal additive genetic effect; and Model 3 included those in 2 plus the permanent maternal environment effect. All of them used the residual maximum likelihood method (REML), implemented in the ASReml prog ram. The best model to obtain variance components and genetic parameters was the second model, direct heritability (h²d ± s. e.) 0.11 ± 0.09 and 0.19 ± 0.18 and maternal (h²m ± s. e.) 0.15 ± 0.06 and 0.14 ± 0.11 for BW and WW, respectively. The direct-maternal genetic correlations were -0.21 ± 0.29, -0.92 ± 0.11 and -0.84 ± 0.20 for BW, WW and 6MW, respectively.
El objetivo de este estudio fue estimar los componentes de varianza genéticos, directos y maternos para características de crecimiento en un rebaño de ciervo rojo (Cervus elaphus scoticus), en Querétaro, México. Se analizó la información de 1994 hasta 2003 consistente en 417 registros de peso al nacimiento (PN), 169 al destete (PD), 168 a los seis meses (P6M) y 172 al año (PA), incluyó identificación de 554 animales con 11 sementales y 107 hembras. Los efectos fijos considerados fueron: sexo, año de nacimiento y número de parto (P < 0.001). Se utilizaron tres modelos mixtos. El Modelo 1 incluyó efectos fijos y efecto genético aditivo directo; el Modelo 2, igual al 1 más el efecto genético aditivo materno; el Modelo 3, igual al 2 más el efecto del ambiente permanente materno, todos usaron el método de máxima verosimilitud restringida (REML), instrumentado en el programa ASREML. El mejor modelo para obtener los componentes de varianzas y los parámetros genéticos fue el 2, heredabilidades directas (h²d ± e. e.) 0.11 ± 0.09 y 0.19 ± 0.18, y materna (h²m± e. e.) 0.15 ± 0.06 y 0.14 ± 0.11 para PN y PD, respectivamente. Las correlaciones genéticas directas-maternas fueron -0.21 ± 0.29, -0.92 ± 0.11 y -0.84 ± 0.20, para PN, PD y P6M, respectivamente.
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A case of tuberculosis is reported in an eight-year-old, male, elk (Cervus elaphus nelsoni). The elk showed severe coughing, respiratory distress, abdominal breathing, anorexia, and severe progressive emaciation in the elk farm. At necropsy, the elk appeared in poor body condition. Mild enlargement of retropharyngeal and submandibular lymph node was observed in the head. Diffuse fibrinous pleuritis and purple red lobar pneumonia were found in the thorax. Well demarcated numerous dark yellow discrete or confluent nodules from 0.3 to 2 cm in diameter were scattered in the whole lung. Bronchial and mediastinal lymph nodes were also enlarged. Histopathologically, lungs had typical classical tuberculous granulomas, multiple abscesses, and numerous macrophages and Langhans giant cells infiltration in alveolar lumen. In the lymph nodes, there were small clusters of necrosis and infiltration of numerous macrophages, epithelioid cells, and Langhans giant cells. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. According to this study, there are differences of the histopathologic lesions and the numbers of acid-fast bacilli in the lesions between this elk and cattle. Mycobacterium bovis was confirmed as a causative agent in this elk using bacterial isolation, biochemical characteristics, and PCR technique. The isolate was negative for niacin test, nitrate reductase, and pyrazinamidase. This is a first report for bovine tuberculosis of farmed elk in Asia.