Diet-Induced Alteration of the Murine Intestinal Microbiome Following Antibiotic Ablation

Abstract

Mouse models of antibiotic-induced ablation of the intestinal microbiome have been used to study the microbiome in health and disease. The fecal microbiomes of mice treated with broad-spectrum antibiotics while being fed different laboratory chows were analyzed by Gram stain, quantitative flow cytometry, bacterial cell culture, next generation sequencing of the V3 regions of the 16S ribosomal RNA (rRNA) gene, microscopy, and sequence analysis of the tuf gene. Noncultivatable gram-positive cocci and cultivatable yeast were the microorganisms most readily detected in feces of antibiotic-treated mice fed a defined diet that utilizes casein as a protein source, maltodextrin 10 and sucrose as sources of carbohydrates, and lard as the major source of fat. High-throughput sequencing of the variable regions of the 16S rRNA gene and tuf gene sequencing identified the major bacterial phylotype as Lactococcus. The mouse chow was heavily laden with noncultivatable Lactococcus, which dominated the intestinal flora after consumption. The microbiome of antibiotic-treated mice fed a grain-based diet (mainly wheat, corn and alfalfa) consisted predominantly of a member of the Enterobacteriaceae identified as Escherichia coli, and yeast was not detected by culture or Gram stain. Appearance of intestinal yeast by culture and Gram stain was dependent on the specific chow, although yeast was not detected by culture or Gram stain in the chow. We conclude that bacteria found in food sources can influence qualitative and quantitative assessments of the fecal microbiome, at least in the 546context of antibiotic therapy, and potentially confound molecular studies that assess the effects of diet on the intestinal ecology. Not surprisingly, different food sources can influence the microbiome, particularly in the context of antibiotic-mediated ablation of the intestinal microbiome. Whether and how the food-derived dead bacteria alter intestinal physiology needs to be determined.

A New Technique for Use in Culturing Prokaryotes Comprising the Mouse Intestinal Microbiome

“…has been shown to grow better in anaerobic conditions [44], leading to increased lactic acid production and lower pH. Lactobacillus has also been shown to be present in commercially-available mouse food, and thus feeding selection may impact its presence or absence in the gastrointestinal microbiome [9]. The presence of oxygen also likely affected species from the genus Enterococcus as it significantly increased between mice cultures and the pooled microbiome, especially those with a lower pH (Figure 7(A), Figure 7(B); Supplemental Table S9).…”

“…The microbiome forms a symbiotic relationship with its host, and impacts metabolism, response to nutrients, and physiological and immunological development [9]. Essentially, microbes have a cooperative role in the GI tract and contribute to a host's immune system and metabolism [10] [11] [12] [13] with Firmicutes and Bacteroidetes being the predominant Gram-positive and Gramnegative phyla, respectively [9].…”

“…The microbiome forms a symbiotic relationship with its host, and impacts metabolism, response to nutrients, and physiological and immunological development [9]. Essentially, microbes have a cooperative role in the GI tract and contribute to a host's immune system and metabolism [10] [11] [12] [13] with Firmicutes and Bacteroidetes being the predominant Gram-positive and Gramnegative phyla, respectively [9]. Although the natural relationship between the microbiome and the host is essential, overpopulation by an undesirable species, or dysbiosis, has been linked to particular diseases and phenomena such as autism spectrum disorder [14], cancer [11], and obesity [15].…”

Administration of Bovine Milk Oligosaccharide to Weaning Gnotobiotic Mice Inoculated with a Simplified Infant Type Microbiota

“…Information on genus was investigated if relevant to the interpretation of results. Sequences assigned to OTU f__Streptococcaeceae;g__Lactococcus were filtered out as they originated from Lactococcus precipitated casein in the diet [ 15 ]. Samples with low DNA concentration (<0.1 ng/µL) were excluded from the abundance analysis.…”