Samsudin Anjas Asmara, Wright André-Denis, Al Jassim Rafat
School of Agriculture and Food Sciences, The University of Queensland, Gatton, QLD, 4343, Australia.
Trop Anim Health Prod. 2014 Oct;46(7):1161-6. doi: 10.1007/s11250-014-0621-6. Epub 2014 Jun 5.
The total bacterial community of Fibrobacter succinogenes and Ruminococcus flavefaciens in fibre-enriched culture of the foregut contents of 12 adult feral camels (Camelus dromedaries) fed on native vegetation in Australia was investigated using quantitative PCR. Foregut contents were collected postmortem, pooled and filtered before divided into two fractions. One fraction was used for extraction of DNA, while the other fraction was inoculated straight away into BM 10 contained filter paper (FP), cotton thread (CT) or neutral detergent fibre (NDF) as the sole carbohydrate sources in Hungate tubes. The tubes were incubated anaerobically at 39 °C for 1 week. After a near complete degradation of the FP and CT and extensive turbidity in the NDF, media subculturing was carried out into fresh media tubes. This was repeated twice before genomic DNA was extracted and used for quantification of bacteria. Using an absolute quantification method, the numbers of cells in 1 ml of each sample ranged from 4.07 × 10(6) to 2.73 × 10(9) for total bacteria, 1.34 × 10(3) to 2.17 × 10(5) for F. succinogenes and 5.78 × 10(1) to 3.53 × 10(4) for R. flavefaciens. The mean cell number of F. succinogenes was highest in the FP enrichment medium at approximately 107-fold, whereas for the R. flavefaciens targeted primer, the NDF enrichment media had the highest mean cell number at approximately 4-fold when compared to the rumen content. The data presented here provide evidence of fibre type preference by the two main fibre-degrading bacteria and would help us understand the interaction between fibre type and fibre-degrading microorganisms, which has ramification on camel nutrition at different seasons and environments.
采用定量PCR技术,对12只以澳大利亚本土植被为食的成年野生骆驼(单峰驼)前肠内容物纤维富集培养物中的琥珀酸纤维杆菌和黄化瘤胃球菌的总细菌群落进行了研究。前肠内容物在骆驼死后采集,合并后过滤,然后分为两部分。一部分用于提取DNA,另一部分立即接种到含有滤纸(FP)、棉线(CT)或中性洗涤纤维(NDF)作为唯一碳水化合物来源的Hungate管中的BM 10培养基中。将试管在39℃下厌氧培养1周。在FP和CT几乎完全降解且NDF出现大量浑浊后,将培养基转接至新鲜的培养基试管中。重复此操作两次,然后提取基因组DNA并用于细菌定量。使用绝对定量方法,每毫升各样本中的细胞数量,总细菌为4.07×10(6)至2.73×10(9),琥珀酸纤维杆菌为1.34×10(3)至2.17×10(5),黄化瘤胃球菌为5.78×10(1)至3.53×10(4)。琥珀酸纤维杆菌的平均细胞数在FP富集培养基中最高,约为107倍;而对于黄化瘤胃球菌靶向引物,与瘤胃内容物相比,NDF富集培养基中的平均细胞数最高,约为4倍。本文提供的数据证明了两种主要纤维降解细菌对纤维类型的偏好,这将有助于我们理解纤维类型与纤维降解微生物之间的相互作用,这对不同季节和环境下骆驼的营养状况具有影响。