RESUMEN
The abilities of certain microorganisms to be transferred across the food production chain, persist in the final product and, potentially, colonize the human gut are poorly understood. Here, we provide strain-level evidence supporting that dairy cattle-associated bacteria can be transferred to the human gut via consumption of Parmesan cheese. We characterize the microbial communities in samples taken from five different locations across the Parmesan cheese production chain, confirming that the final product contains microorganisms derived from cattle gut, milk, and the nearby environment. In addition, we carry out a human pilot study showing that Bifidobacterium mongoliense strains from cheese can transiently colonize the human gut, a process that can be enhanced by cow milk consumption.
Asunto(s)
Queso/microbiología , ADN Bacteriano/genética , Microbioma Gastrointestinal/genética , Leche/microbiología , Filogenia , Animales , Bifidobacterium/clasificación , Bifidobacterium/genética , Bifidobacterium/aislamiento & purificación , Bovinos , Corynebacterium/clasificación , Corynebacterium/genética , Corynebacterium/aislamiento & purificación , Código de Barras del ADN Taxonómico , Heces/microbiología , Humanos , Lactobacillus delbrueckii/clasificación , Lactobacillus delbrueckii/genética , Lactobacillus delbrueckii/aislamiento & purificación , Proyectos Piloto , Prevotella ruminicola/clasificación , Prevotella ruminicola/genética , Prevotella ruminicola/aislamiento & purificación , ARN Ribosómico 16S/genética , Streptococcus thermophilus/clasificación , Streptococcus thermophilus/genética , Streptococcus thermophilus/aislamiento & purificaciónRESUMEN
AIM: To reassemble Prevotella ruminicola genome from rumen metagenomic data of cattle and buffalo and compare with the published reference genome. METHOD: Rumen microbial communities from Mehsani buffaloes (n = 8) and Kankrej cattle (n = 8), each adapted to different proportions of a dry or green roughage diet, were subjected to metagenomic sequencing by Ion Torrent PGM, and subsequent reads were analyzed by MG-RAST. Using reference-guided assembly of the sequences against the published P. ruminicola strain 23, draft genomes of 2.56 and 2.46 Mb were reconstructed from Mehsani buffalo and Kankrej cows, respectively. The genomes were annotated using the RAST Server and carbohydrate active enzyme (CAZyme) analysis. RESULTS: Taxonomic analysis by MG-RAST revealed P. ruminicola to be the most abundant species present among the rumen microflora. Functional annotation of reconstructed genomes using the RAST Server depicted the maximum assignment of coding sequences involved in the subsystems amino acid and derivatives and carbohydrate metabolism. CAZyme profiling revealed the glycoside hydrolases (GH) family to be the most abundant. GH family subclassification revealed that the extracted genomes had more sequence hits for GH2, GH3, GH92 and GH97 as compared to the reference. CONCLUSION: The results reflect the metabolic significance of rumen-adapted P. ruminicola in utilizing a coarse diet for animals based on acquisition of novel genetic elements.
Asunto(s)
Prevotella ruminicola/genética , Rumen/microbiología , Animales , Bacterias/clasificación , Bacterias/enzimología , Bacterias/genética , Bacterias/aislamiento & purificación , Proteínas Bacterianas/genética , Búfalos , Bovinos , Microbioma Gastrointestinal , Genoma Bacteriano , Metagenómica , Sistemas de Lectura Abierta , Filogenia , Prevotella ruminicola/clasificación , Prevotella ruminicola/enzimología , Prevotella ruminicola/aislamiento & purificaciónRESUMEN
The Prevotellas comprise a diverse group of bacteria that has received surprisingly limited attention at the whole genome-sequencing level. In this communication, we present the comparative analysis of the genomes of Prevotella ruminicola 23 (GenBank: CP002006) and Prevotella bryantii B(1)4 (GenBank: ADWO00000000), two gastrointestinal isolates. Both P. ruminicola and P. bryantii have acquired an extensive repertoire of glycoside hydrolases that are targeted towards non-cellulosic polysaccharides, especially GH43 bifunctional enzymes. Our analysis demonstrates the diversity of this genus. The results from these analyses highlight their role in the gastrointestinal tract, and provide a template for additional work on genetic characterization of these species.
Asunto(s)
Genoma Bacteriano , Prevotella ruminicola/genética , Prevotella/genética , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Datos de Secuencia Molecular , Filogenia , Prevotella/clasificación , Prevotella/enzimología , Prevotella/aislamiento & purificación , Prevotella ruminicola/clasificación , Prevotella ruminicola/enzimología , Prevotella ruminicola/aislamiento & purificación , Rumen/microbiologíaRESUMEN
A real-time PCR approach was used in this study to clarify the populations of major bacterial species in the rumens of faunated and unfaunated cattle. The sensitivity of this novel real-time PCR assay was evaluated by using 10(1) to 10(8) plasmid copies of target bacteria. The numbers of plasmid copies of Ruminococcus albus, Ruminococcus flavefaciens, Prevotella ruminicola, and the CUR-E cluster were higher in the unfaunated than in the faunated rumens. The CUR-E cluster belongs to the Clostridium group. In contrast, Fibrobacter succinogenes was higher in the faunated than in the unfaunated rumens. Although it is well known that an absence of protozoa brings about an increase in the bacterial population, it was clarified here that an absence of protozoa exerted differential effects on the populations of cellulolytic bacteria in cattle rumens (i.e., F. succinogenes, R. albus, and R. flavefaciens). In addition, real-time PCR analysis suggested that the CUR-E cluster was more prevalent in the unfaunated rumens.