共查询到20条相似文献,搜索用时 0 毫秒
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A method for the separation of leucocytes from bovine blood and its separation into lymphocyte and granulocyte fractions is described. The method, involving flash lysis of the erythrocyte population, was found to yield large numbers of viable cells suitable for maintenance in tissue culture medium and hence of value in immunological studies. 相似文献
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Luzia Teixeira Raquel M. Marques Artur P. Águas Paula G. Ferreira 《Research in veterinary science》2011,91(1):164-166
Rabbit Haemorrhagic Disease Virus (RHDV), a member of the Caliciviridae family, is the etiologic agent of Rabbit Haemorrhagic Disease (RHD); this viral disease is highly contagious and kills more than 90% of infected adult rabbits. Research on experimental calicivirus infection uses inocula obtained from livers of rabbits dying from calicivirus infection. This implies that caliciviruses have to be purified from liver homogenates. Current methods to isolate caliciviruses from rabbit livers are time consuming. We propose here a new procedure for fast purification of rabbit caliciviruses from liver homogenates that uses centrifugation through an iodixanol gradient. This method offers in approximately 2 h a sample with a high degree of calicivirus purity, as shown by its biochemical and immunocytochemistry analysis, which is also able to kill adult rabbits from RHD within 48 h of inoculation. 相似文献
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Total respiratory impedance was measured rapidly and noninvasively in conscious horses over the frequency range 3 to 40 Hz by the forced random noise method. The shape of curve of impedance versus frequency in horses was markedly different from that of humans. Respiratory resistance was readily found as the real part of impedence and both its absolute value and frequency dependence are useful indices of pulmonary function. It was difficult to obtain meaningful results in intubated animals with the method because of the mechanical properties of the endotracheal tube itself. 相似文献
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P C Simon 《Canadian journal of veterinary research》1975,39(3):349-353
A hemagglutination inhibition test for the rapid identification of Sphaerophorus necrophorus is described. Erythrocytes from six species of animals were tested and human cells were found to be the best agglutination indicators. Antiserum prepared in rabbits was found to be specific for S. necrophorus hemagglutinins when tested against 20 isolates of S. necrophorus and 117 other bacteria belonging to 22 genera. The possibility of using a hemagglutination inhibition test for the detection of bovine necrobacillosis was explored. 相似文献
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A simple, rapid microtechnique utilizing whole blood has been developed for evaluating the in vitro stimulation of canine lymphocytes. The test uses 5 ul. of heparinized whole blood per culture in a microtiter plate and requires no serum supplement. Cultures are harvested by an automated multiple sample cell harvester. This technique permits large numbers of replicate samples to be tested on individual animals with minimal amounts of blood and minimal technician time. 相似文献
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Optimal conditions for stimulation by phytohaemagglutinin (PHA) were established for equine, porcine, ovine and human lymphocytes in MEMS medium. Optimal thymidine concentration was determined for assay of cell transformation. With all species tested horse serum gave highest thymidine incorporation. Homologous serum was not more appropriate for lymphocytes of man, pig and sheep. Optimal stimulation was achieved at 20, 0.5–5, 5, and 10–40 μg PHA per 106 cells for human, equine, porcine and ovine lymphocytes, respectively. 相似文献
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A whole blood method for measuring mitogen-induced transformation of sheep lymphocytes 总被引:1,自引:0,他引:1
H J Larsen 《Research in veterinary science》1979,27(3):334-338
A simple and reproducible micromethod for determination of in vitro mitogenic responses of sheep peripheral blood lymphocytes is described. The test uses (i) whole blood diluted in RPMI 1640 medium to give a cell count of 0.5 x 106-1 x 106 lymphocytes/ml, (ii) mitogens in the range of 5-20 micrograms of phytohaemagglutinin/ml, 20-80 micrograms of lipopolysaccharide/ml or 20-80 microliters of poke weed mitogen/ml, and (iii) a stimulation time of 42 to 90 h. A considerable variation in mitogenic response was observed both between animals and on different occasions in the same animal. Because of the large periodic variation it was suggested that the test should be repeated using blood drawn at different times in order to determine the mitogenic response of an animal. 相似文献
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Goto-Koshino Y Ohno K Nakajima M Mochizuki H Kanemoto H Tsujimoto H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(6):773-778
Macrophages play an important role in a variety of situations, including pathogen elimination, inflammation, and tissue repair. However, these cells are not fully studied in dogs, in part, due to the difficulty of efficiently isolating and culturing them in vitro. In this study, we cultured canine peripheral blood mononuclear cells (PBMCs) with 10 ng/ml of phorbol 12-myristate-13-acetate (PMA) for 5 days to obtain macrophages. A high number of round-adherent cells were obtained without the addition of any cytokine. These cells showed active phagocytic activity and a cell surface antigen profile different from dendritic cells. Our method facilitates a high yield of macrophages in a short cultivation period compared with previous studies. This method might be a powerful tool to study macrophage functions in dogs. 相似文献
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A simple, rapid and reliable method for selecting or assessing the number of replicates for animal experiments 总被引:1,自引:0,他引:1
W E Berndtson 《Journal of animal science》1991,69(1):67-76
A simple approach was developed for determining the number of replicates needed per treatment group to provide experiments of known power and sensitivity, where power equals the probability that a treatment effect would not go undetected if an effect existed and sensitivity equals the minimal treatment response that will be detectable. This approach, in turn, was used to construct reference tables, applicable across scientific disciplines, from which researchers may read replication requirements directly with ease, speed and reliability. To use the tables, one need only furnish a reliable estimate of the coefficient of variability expected among replicates, which may be obtained from prior observations on similar populations. The tabular data also enable a rapid, reliable assessment of the actual power and sensitivity of completed experiments, such as those contained within the published literature. 相似文献
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Various methods have been developed for separation of the cellular components of blood. Size and density differences in cellular blood elements of various animal species necessitate the use of specific separative procedures. This study describes a method which combines isopycnic density-gradient centrifugation with erythrocyte aggregation as a means of isolating leukocytes from bovine blood. The procedure yields a viable population of mixed leukocytes which has proven useful for cell culture and in vitro tests of cell-mediated immunity. 相似文献
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J R Stabel T L Bosworth T A Kirkbride R L Forde R H Whitlock 《Journal of veterinary diagnostic investigation》2004,16(1):22-30
Diagnosis of paratuberculosis (Johne's disease) is stymied by the lack of 1 diagnostic tool that can be used to detect both subclinically and clinically infected animals. At present, fecal culture remains the single diagnostic test that can detect infection in both disease states provided the animals actively shed Mycobacterium paratuberculosis in their feces. Yet, fecal culture has a disadvantage associated with the protracted incubation period of 8-16 weeks before results are available. Detection of nucleic acids specific to M. paratuberculosis in fecal samples is a technique that can circumvent the culture method. This study describes a rapid, simple, and effective method to extract DNA from fecal samples and modification of a polymerase chain reaction assay for optimal sensitivity of detection. An evaluation of 1,000 well-characterized fecal samples was performed by the Colorado Department of Agriculture (Denver, CO) and the National Animal Disease Center (Ames, IA) to determine the sensitivity, specificity, and reproducibility of the new method. Results from this study show that the sensitivity of detection was highly dependent on the load of bacteria in the fecal sample with 81% detection of samples containing >70 colony-forming units (cfu)/g of feces and a 45% detection rate for samples containing less than 1 cfu/g. Similarly, reproducibility of the technique between the 2 laboratories (n = 250 samples) was much higher (75%) for the fecal samples containing high levels of M. paratuberculosis and reduced to 25% for samples with less than 1 cfu/g. An overall specificity of 83% was obtained for known negative samples. The method described here is rapid, simple, and inexpensive compared with other techniques. In addition, this method can detect animals that are shedding less than 1 cfu/g. 相似文献
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