Craig A M, Latham C J, Blythe L L, Schmotzer W B, O'Connor O A
College of Veterinary Medicine, Oregon State University, Corvallis 97331-4802.
Appl Environ Microbiol. 1992 Sep;58(9):2730-6. doi: 10.1128/aem.58.9.2730-2736.1992.
The ability of ovine ruminal fluid to metabolize pyrrolizidine alkaloid (PA) from Senecio jacobaea under anaerobic conditions was evaluated. Four fistulated sheep fed PA served as individual sources of ruminal fluid, which was incubated in a defined minimal salts medium under two different anaerobic conditions, denitrifying and methanogenic. Anaerobic cultures amended with ovine ruminal fluids (20%), PA (100 micrograms/ml), and a defined minimal salts medium were monitored for a period of several days. These cultures revealed that while PA was not depleted in sterile, autoclaved controls or under denitrifying conditions, it was metabolized during periods of active methanogenesis under methanogenic conditions. In addition, samples of ruminal fluid were separated by differential centrifugation under anaerobic conditions, and the resultant supernatants were tested for their ability to metabolize PA as compared with those of the respective uncentrifuged control fluids. Uncentrifuged controls exhibited a PA depletion rate of -4.04 +/- 0.17 micrograms of PA per ml per h. Supernatants 1 (centrifuged at 41 x g for 2 min), 2 (centrifuged at 166 x g for 5 min), and 3 (centrifuged at 1,500 x g for 10 min) exhibited significantly slower depletion rates, with slopes of data representing -1.64 +/- 0.16, -1.44 +/- 0.16, and -1.48 +/- 0.16 micrograms of PA metabolized per ml per h, respectively, demonstrating no statistically significant difference among the supernatant cultures. Microscopic evaluations revealed that protozoa were present in the control whole ruminal fluid and to a lesser extent in supernatant 1, while supernatants 2 and 3 contained only bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)
评估了绵羊瘤胃液在厌氧条件下代谢千里光中的吡咯里西啶生物碱(PA)的能力。四只饲喂PA的带瘘管绵羊作为瘤胃液的个体来源,将瘤胃液在特定的基本盐培养基中于两种不同的厌氧条件下培养,即反硝化条件和产甲烷条件。用绵羊瘤胃液(20%)、PA(100微克/毫升)和特定的基本盐培养基改良的厌氧培养物监测了几天。这些培养表明,虽然在无菌、高压灭菌的对照中或反硝化条件下PA没有消耗,但在产甲烷条件下活跃产甲烷期间它被代谢了。此外,在厌氧条件下通过差速离心分离瘤胃液样本,并将所得上清液与相应未离心的对照液相比,测试其代谢PA的能力。未离心的对照液显示PA消耗速率为每毫升每小时-4.04±0.17微克PA。上清液1(以41×g离心2分钟)、2(以166×g离心5分钟)和3(以1500×g离心10分钟)的消耗速率明显较慢,数据斜率分别代表每毫升每小时代谢-1.64±0.16、-1.44±0.16和-1.48±0.16微克PA,表明上清液培养物之间无统计学显著差异。显微镜评估显示,原生动物存在于对照全瘤胃液中,在上清液1中程度较轻,而上清液2和3仅含有细菌。(摘要截断于250字)