Isolation of previously uncultured rumen bacteria by dilution to extinction using a new liquid culture medium

Kenters, Nikki; Henderson, Gemma; Jeyanathan, Jeyamalar; Kittelmann, Sandra; Janssen, Peter H.

doi:10.1016/j.mimet.2010.10.011

Cited by 91 publications

(90 citation statements)

References 40 publications

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“…In the traditional isolation process, MCs were supplied as the sole carbon and nitrogen source (Valeria et al, 2006;Zhang et al, 2010), resulting in a quick elimination of MCs and enrichment of the target bacteria. The longer growth period and fewer colonies species in the pore water and concentrated water extract of sediment may be due to the extra nitrogen and carbon source (Table 1), which agreed well with the report that exogenous nutrients led to slow bacteria growth (Kenters et al, 2011;Markiewicz et al, 2011). It was hard to prepare natural media with the equivalent nutrients level as artificial media during microorganism isolation.…”

Section: Influence Of Nutrient Condition On Bacteria Isolationsupporting

confidence: 83%

“…In our study, no colonies were shown with direct application of water extracts of sediment (10%, w/v) after 20 days (Table 2), evaluations of the nutrient condition revealed that the sediment extraction was characterized by 1.9 mg L − 1 TN and rather low or even missed mineral salts (Table 1). It was recognized that microorganism from the environment had a requirement for components from the media (Kenters et al, 2011). It might be the poor nutrient condition that was responsible for the failure of bacteria cultivation through water extracts of sediment extracts.…”

Section: Influence Of Nutrient Condition On Bacteria Isolationmentioning

confidence: 99%

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Enhanced and continued degradation of microcystins using microorganisms obtained through natural media

Hong

Pan

2014

Journal of Microbiological Methods

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Microorganisms isolated through artificial media are often unsustainable in biodegrading microcystins (MCs) in natural water. Here we studied alternative approaches to isolate MCs-degrading bacteria using natural media. In comparison to two species (MS-1 and MS-2) isolated from artificial media and the failure of bacterial colonies formation using water extracts of sediment (10%, w/v), five colony species (WC-1 to WC-5) appeared using concentrated water extracts of sediment that is 10-fold enhancement of nutrient level. In the simulated biodegradation test in Lake Taihu water with continuous supply of MCs, a lag phase of 6 days was required for MS-1 and M-2 to degrade 13% and 15% of the added MC-RR and MC-LR, respectively, whereas the lag phase was only 3 days with approximately 44% and 31% removal of the added MC-RR and MC-LR by WC-1 to WC-5. During the continuous supply experiment, degradation of MCs by MS-1 and MS-2 stopped after 3 days, while degradation of MCs by WC-1 to WC-5 lasted continuously throughout the 18 day test period with 2 to 6-fold enhancement of removal rate. 16S rRNA gene sequences and phylogenetic analysis indicated the potential to amplify species of MCsdegrading bacteria when natural media were used. The results suggested that the increased adaptability of bacteria obtained through concentrated natural media was responsible for the enhanced and continued biodegradation under simulated natural water conditions.

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Section: Influence Of Nutrient Condition On Bacteria Isolationsupporting

confidence: 83%

Section: Influence Of Nutrient Condition On Bacteria Isolationmentioning

confidence: 99%

Enhanced and continued degradation of microcystins using microorganisms obtained through natural media

Hong

Pan

2014

Journal of Microbiological Methods

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“…The putative class III bacteriocins in R. albus AD2013 and R. albus SY3 had identical amino acid sequences. R. albus strain SY3 was isolated from anaerobic cellulose roll tubes (53), and R. albus AD2013 was obtained from the rumen of a New Zealand cow as part of a study to isolate previously uncultured ruminal bacteria (54).…”

Section: Discussionmentioning

confidence: 99%

Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes

Azevedo

Bento

Ruiz

et al. 2015

Appl Environ Microbiol

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b Some species of ruminal bacteria are known to produce antimicrobial peptides, but the screening procedures have mostly been based on in vitro assays using standardized methods. Recent sequencing efforts have made available the genome sequences of hundreds of ruminal microorganisms. In this work, we performed genome mining of the complete and partial genome sequences of 224 ruminal bacteria and 5 ruminal archaea to determine the distribution and diversity of bacteriocin gene clusters. A total of 46 bacteriocin gene clusters were identified in 33 strains of ruminal bacteria. Twenty gene clusters were related to lanthipeptide biosynthesis, while 11 gene clusters were associated with sactipeptide production, 7 gene clusters were associated with class II bacteriocin production, and 8 gene clusters were associated with class III bacteriocin production. The frequency of strains whose genomes encode putative antimicrobial peptide precursors was 14.4%. Clusters related to the production of sactipeptides were identified for the first time among ruminal bacteria. BLAST analysis indicated that the majority of the gene clusters (88%) encoding putative lanthipeptides contained all the essential genes required for lanthipeptide biosynthesis. Most strains of Streptococcus (66.6%) harbored complete lanthipeptide gene clusters, in addition to an open reading frame encoding a putative class II bacteriocin. Albusin B-like proteins were found in 100% of the Ruminococcus albus strains screened in this study. The in silico analysis provided evidence of novel biosynthetic gene clusters in bacterial species not previously related to bacteriocin production, suggesting that the rumen microbiota represents an underexplored source of antimicrobial peptides.T he production of low-molecular-weight antimicrobial peptides is a trait widely distributed among species of bacteria and archaea (1). Although a variety of functions has been assigned to these compounds (e.g., toxins, virulence factors, bacterial hormones), the bacteriocins have mainly been investigated due to their potential as alternatives to antibiotics and food preservatives (2-4). With the rise in antibiotic resistance among commensal and pathogenic strains of bacteria, there is an urgent need to identify and develop novel therapeutic strategies for clinical applications in human and animal health (5).Bacteriocins are often defined as ribosomally synthesized antimicrobial peptides produced by bacteria or archaea (6). Bacteriocins show great diversity in their chemical structures and mechanisms of action, and at least four main groups have been proposed to classify these antimicrobial agents (7). Class I contains posttranslationally modified antimicrobial peptides, such as lanthipeptides, sactipeptides, and lasso peptides, which differ in their molecular structures, their mechanisms of action, and the enzymatic apparatuses involved in modifying the precursor peptides (7,8). Class II bacteriocins consist of antimicrobial peptides without posttranslationally modified residues and ca...

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“…Effect of gum arabic on growth of B. fibrisolvens and B. proteoclasticus B. fibrisolvens D1 (DSM 3071) and B. proteoclasticus P18 (Rowett Institute of Nutrition and Health) were grown anaerobically for 48 h at 39°C in medium 704 (DSMZ) or RM02 medium (Kenters et al, 2011) in 12.5 × 1.5 cm culture tubes closed with screw caps fitted with butyl rubber septa (Bellco Biotechnology, Vineland, NJ, USA). The medium 704 was modified by omitting the VFA mixture, haemin and glycerol, and by increasing the proportion of rumen fluid to 20% (v/v).…”

Section: Distribution Of 22:6n-3 In Incubation Contentsmentioning

confidence: 99%

Effect of adsorbants on in vitro biohydrogenation of 22:6n-3 by mixed cultures of rumen microorganisms

Vlaeminck

Jeyanathan

Thanh

et al. 2016

animal

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Studies on microbial biohydrogenation of fatty acids in the rumen are of importance as this process lowers the availability of nutritionally beneficial unsaturated fatty acids for incorporation into meat and milk but also might result in the accumulation of biologically active intermediates. The impact was studied of adsorption of 22:6n-3 (DHA) to particulate material on its disappearance during 24 h in vitro batch incubations with rumen inoculum. Four adsorbants were used in two doses (1 and 5 mg/ml of mucin, gum arabic, bentonite or silicic acid). In addition, the distribution of 22:6n-3 in the pellet and supernatant of diluted rumen fluid was measured. Bentonite and silicic acid did not alter the distribution of 22:6n-3 between pellet and supernatant nor increased the disappearance of 22:6n-3 during the incubation. Both mucin and gum arabic increased the recovery of 22:6n-3 in the supernatant, indicating that these compounds lowered the adsorption of the fatty acid to ruminal particles. This was associated with an increased disappearance of 22:6n-3, when initial 22:6n-3 was 0.06 or 0.10 mg/ml, and an increased formation of 22:0, when initial 22:6n-3 was 0.02 mg/ml, during the 24 h batch culture experiment. Addition of gum arabic to pure cultures of Butyrivibrio fibrisolvens or Butyrivibrio proteoclasticus did not negate the inhibitory effect of 22:6n-3 on growth. As both mucin and gum arabic provide fermentable substrate for ruminal bacteria, an additional experiment was performed in which mucin and gum arabic were replaced by equal amounts of starch, cellulose or xylan. No differences in disappearance of 22:6n-3 were observed, suggesting that the stimulatory effect of mucin and gum arabic on disappearance of 22:6n-3 most probably is not due to provision of an alternative site of adsorption but related to stimulation of bacterial growth. A relatively high proportion of 22:6n-3 can be reduced to 22:0 provided the initial concentration is low.

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Isolation of previously uncultured rumen bacteria by dilution to extinction using a new liquid culture medium

Cited by 91 publications

References 40 publications

Enhanced and continued degradation of microcystins using microorganisms obtained through natural media

Enhanced and continued degradation of microcystins using microorganisms obtained through natural media

Distribution and Genetic Diversity of Bacteriocin Gene Clusters in Rumen Microbial Genomes

Effect of adsorbants on in vitro biohydrogenation of 22:6n-3 by mixed cultures of rumen microorganisms

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