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ABSTRACT: To compare the fermentation characteristics of D-lactate, L-lactate and DL-lactate mixture by fecal microbiota of pigs, in vitro fermentation test was conducted with D-lactate, L-lactate and DL-lactate mixture as substrates, feces of piglets as inocula. The concentrations of lactate and short chain fatty acid (SCFA) were quantified during the fermentation. The composition of bacterial communities in the inocula and 24 h fermentation fluids was analyzed by pyrosequencing. The results showed that, when 5 mmol/l lactate was used, there was no significant difference in utilizing efficiency among D-lactate, L-lactate and DL-lactate mixture, propionate and butyrate were the major end-products converted from lactate. However, when 25 mmol/l lactates were used, a higher utilizing efficiency of DL-lactate mixture and a slower utilizing rate of D-lactate were observed, acetate and propionate became the main end-products. The SCFA proportions were significantly affected by lactate source and its concentration. Pyrosequencing analysis showed that after 24 h fermentation, significant difference in the composition of bacterial communities (genus and OTU levels) was observed among different substrate groups. The results suggest that lactate accumulation in the hindgut is related to the cooperation between microbiota members, and regulating bacterial community may be a possible way to control lactate accumulation.
Anaerobe 11/2012; · 2.41 Impact Factor
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ABSTRACT: The diversity of fecal methanogens of Erhualian (obese type) and Landrace (lean type) pigs was examined using separate 16S rRNA gene libraries for each breed. A total of 763 clones were analyzed; 381 from the Erhualian library and 382 from the Landrace library were identified belonging to the genus Methanobrevibacter. Others were identified belonging to the genus Methanosphaera. The two libraries showed significant differences in diversity (P < 0.05) and composition (P < 0.0001). Only two operational taxonomic units (OTUs) were found in both libraries, whereas six OTUs were found only in the Erhualian library and 23 OTUs were found only in the Landrace library. Real-time PCR showed that the abundance of fecal methanogens in Landrace pigs was significantly higher than that in Erhualian pigs (P < 0.05). Results showed that the Landrace pig (lean) harbored a greater diversity and higher numbers of methanogen mcrA gene copies than the Erhualian pig (obese). These differences may be related to the fatness or leanness in these two pig breeds. The results provide new leads for further investigations on the fat storage of pigs or even humans.
Archaea 01/2012; 2012:605289.
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ABSTRACT: To compare volatile fatty acid (VFA) concentration and microbiota in faeces between healthy and diarrhoeal piglets.
Fecal samples from healthy and diarrhoeal piglets were collected. VFA concentration was determined by GC analysis. Total bacterial DNA was extracted and used for molecular analysis of microbiota.
As compared to healthy piglets, the concentration of acetate in feces of diarrhoeal piglets tended to increase, while branched chain fatty acid (BCFA) decreased significantly (p < 0.05), total VFA (TVFA), propionate and butyrate tended to decrease. The ratio of acetate to TVFA in faeces of diarrhoeal piglets was significantly higher than that in healthy piglets (p < 0.05), while the ratio of propionate and BFVA to TVFA decreased significantly after diarrhoea (p < 0.05). DGGE analysis of total bacterial community and Clostridium cluster IV group showed no significant changes in both bacterial community were found in faeces of piglets after diarrhoea. Similarity analysis of DGGE profiles revealed that faecal samples of diarrhoeal piglets tended to gather in the same cluster. Real-time PCR results showed that as compared to healthy piglets, the 16S rRNA gene copies of total bacteria and Clostridium cluster IV decreased significantly in faeces of piglets after Diarrhea (p < 0.05), while there was no significant change in the numbers of E coli and Lactobacilli.
VFA composition in faeces of diarrhoeal piglets changed accompanying with the shift of microbiota as compared to healthy piglets.
ACTA MICROBIOLOGICA SINICA 12/2011; 51(12):1632-8.
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ABSTRACT: We investigated the correlation between the methanogen diversity in the piglet colon and the environmental factor (weaning stress, diet type and age).
The colonic contents of piglets at 7, 14, 21, 28 and 35 days old were collected for the determination of volatile fatty acids and total DNA extraction. DNA was subject to PCR-DGGE (Denaturing gradient gel electrophoresis) analysis and interesting bands were excised and sequenced. Redundancy analysis (RDA) was performed for the correlation between methanogen diversity and environmental factors.
As the piglets grew, acetate, propionate and the total volatile fatty acid production increased significantly (P < 0.05), but butyrate concentration remained stable (P > 0.05). The similarity indices of DGGE profiles was higher during the period of the 7 day to 24 day after birth, with grouped in one cluster, and the samples from the 35 days were dropped into another cluster. DGGE analysis showed three dominant bands appeared in samples of 35 d old, with their 16S rRNA gene sequences closely related to Methanobacteria and a novel group of uncultivated Achaean. Redundancy analysis (RDA) showed that age has the largest relevance to the community structure of bacteria in colonic samples, followed by the diet type.
The results indicated that the methanogen community in the piglet colon was stable during the first 24 days after birth, and became more diverse at 35 days of age. The methanogens community was mainly affected by age and the diet type.
ACTA MICROBIOLOGICA SINICA 10/2011; 51(10):1390-7.
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ABSTRACT: To investigate the change of Clostridium cluster IV community in the colon of piglets from 7 to 35 days of age, and its correlation with butyrate concentration.
Three litters of neonatal piglets were used. One piglet from each litter was sacrificed randomly at the age of 7, 14, 21 (weaning day), 24 and 35 days, digestive samples in the colon were collected. The concentration of volatile fatty acid (VFA) was determined by gas chromatography. 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE) and real-time PCR were used for qualitative and quantitative analysis of Clostridium cluster IV community.
Similarity analysis of DGGE profile revealed that samples from piglets at the age of 7 days formed a coherent cluster with indices above 90%, no significant changes in Clostridium cluster IV community were found around weaning period. Real-time PCR analysis showed that 16S rRNA gene copies of total bacteria in the colon of piglets decreased significantly 3 days after weaning, this tendency was in accordance with the changes in concentration of total VFA and butyrate in colon, while there was no significant difference in copies of Clostridium cluster IV group. Sequencing analysis indicated that Clostridium cluster IV group in the colon of piglets were dominated by Faecalibacterium prausnitzii, Subdoligranulum variabile and other uncultured bacteria.
Changes in Clostridium cluster IV community in the colon of piglets were found from the age of 7 days to 14 days, while there was no significant difference during the weaning transition.
ACTA MICROBIOLOGICA SINICA 10/2010; 50(10):1373-9.
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ABSTRACT: A lactate-utilizing, propionate-producing bacterium, strain L9, was isolated from rumen of goat fed with high concentrate by utilizing modified Hungate technique and anaerobic culture technique. The effect of the strain L9 culture on the rumen fermentation was further studied.
According to the characteristics of morphology, physiology, biochemistry tests and sequence comparison of 16S rRNA gene, strain L9 was identified as selenomonas ruminantium. The influence of strain L9 culture on in vitro rumen fermentation was studied using mixed rumen micro-organisms of goats as inoculums.
The results of the metabolism experiment showed that it was capable of using lactate as the sole carbon source, and 90 mmol/L lactate in LH medium could be completely utilized after 24 h incubation. As compared with the control, strain L9 culture addition significantly increased the total volatile fatty acids (TVFA), the percentage of propionate and pH value, while reduced the ratio of acetate to propionate and lactate production (P < 0.05).
The results suggested that strain L9 can reduce lactic acid production and enhance the TVFA and propionate production in in vitro fermentation, and thus could be beneficial for the fermentation of rumen microorganisms.
ACTA MICROBIOLOGICA SINICA 12/2008; 48(12):1571-7.
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ABSTRACT: This present study investigated the changes in bacterial community composition, with an emphasis on Lactobacillus spp. and Streptococcus suis populations as potentially beneficial and harmful groups, in the stomach, jejunum and ileum of piglets after weaning (21 days postpartum) by 16S rRNA gene-based methods. Denaturing gradient gel electrophoresis analysis showed that, after weaning, predominant bands related to Lactobacillus spp. disappeared and were replaced by potential pathogenic species, such as Peptostreptococcus anaerobius, Moraxella cuniculi, S. suis and Porphyromonas catoniae. Real-time PCR revealed that the abundances of lactobacilli and Lactobacillus sobrius as a proportion of total bacterial abundance were significantly lower in the stomach, jejunum and ileum of weaned piglets than in 21-day-old piglets. A specific and sensitive real-time PCR assay was developed for quantification of the important pathogen S. suis within gastrointestinal microbiota. The assay showed that S. suis predominated in the stomach samples of weaned piglets with population levels up to 10(7) copies g(-1) digesta, while it was not detected in the stomach before weaning. Streptococcus suis was not dominant in the jejunum and ileum digesta before weaning, but became dominant after weaning, with population levels up to 10(7) copies g(-1) digesta. The results demonstrated for the first time the postweaning dominance of the potentially harmful S. suis in piglet intestine. The results also suggest that the defensive barrier of the stomach can be impaired as S. suis became dominant while the proportion of Lactobacillus populations decreased after weaning, which may further result in an increase of S. suis abundance in the intestine.
FEMS Microbiology Ecology 07/2008; 66(3):546-55. · 3.41 Impact Factor
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ABSTRACT: To identify Lactobacillus sp. strain S1 from the intestine of piglet, and compare the genomic difference between this strain and Lactobacillus sobrius 001T.
Analysis of 16S rRNA gene sequence and species-specific polymerase chain reaction assay were used to identify Lactobacillus sp. strain S1. Representational difference analysis (RDA) was used to compare these two strains.
The 16S rRNA gene sequence analysis of strain S1 showed that its closest known species in the GenBank database was L. sobrius. Denaturing gradient gel electrophoresis analysis showed that the predominant bands in profiles from bacteria in the jejunum and ileum of piglets had the identical migrating position with strain S1. Cloning and sequencing of 16S rRNA gene analysis revealed that this band matched clone (Clone S) was also related closely to L. sobrius. Phylogenetic analysis of 16S rRNA gene showed that homology between strain S1 and Clone S was 99.8%, and that between strain S1 and L. sobrius was 99.6%. Both strains could be detected in 1.2% agarose gel by L. sobrius-specific PCR assay. Recently, RDA has been adapted to study the genomic diversity of bacterial strains. This analysis showed that there was no genomic difference between the two strains.
Piglet-derived strain S1 belonged to L. sobrius. Piglet-derived L. sobrius 001T and L. sobrius S1 were the similar strain.
ACTA MICROBIOLOGICA SINICA 05/2008; 48(5):577-82.
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ABSTRACT: 16S ribosomal RNA (rRNA) gene based PCR/denaturing gradient gel electrophoresis (DGGE) and real-time PCR were used to monitor the changes in the composition of microbiota in the hindgut of piglets after oral administration of Lactobacillus sobrius S1. Six litters of neonatal piglets were divided randomly into control group and treatment group. At 7, 9, and 11 days of age, piglets in the treatment group orally received a preparation of L. sobrius S1. At 7, 14, 21(weaning), 24, and 35 days of age, one piglet from each litter was sacrificed and digesta samples of hindgut were collected. DGGE analysis of 16S rRNA gene V6-V8 region for all bacteria showed that several populations present in the hindgut of piglets, represented by far-migrating bands, disappeared after weaning. Most of these bands corresponded to Lactobacillus spp. as revealed by sequence analysis. Quantitative real-time PCR specific for lactobacilli further demonstrated that the number of lactobacilli population tended to decrease after the piglets were weaned. Drastic changes of L. amylovorus and L. sobrius in total Lactobacillus populations were also observed in the colon of piglets around weaning, as monitored by 16S rRNA gene V2-V3 region based Lactobacillus-specific PCR-DGGE. Species-specific real-time PCR also revealed that the population of L. sobrius declined apparently in the colon of piglets after weaning. No remarkable changes in the overall microbial community in the hindgut were found between control and treatment groups. However, comparison of DGGE profiles between the two groups revealed a specific band related to Clostridium disporicum that was found in treatment group on day 14. On day 35, a specific band appeared only in the control group, representing a population most closely related to Streptococcus suis (99%). Real-time PCR showed that L. sobrius 16S rRNA gene copies in treatment group were relatively higher than in the control group (10(8.45) vs. 10(6.83)) on day 35, but no significant difference was observed between the two groups.
Anaerobe 05/2008; 14(2):78-86. · 2.41 Impact Factor
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ABSTRACT: Changes of bacterial flora from hindguts of piglets from 7 to 35 days of age (two weeks after weaning) were studied after oral administration of L. amylovorus S1, using molecular techniques based on 16S rDNA gene. Six litters of neonatal piglets were divided randomly into control group and treatment group. At 7, 9, 11 days of age, piglets in treatment group received 1, 2 and 3mL preparation of S1 (5 x 10(9) CFU/mL) through oral administration, respectively. On D 7, 14, 21, 24 and 35, one piglet from each litter was slaughtered and samples of hindguts were collected for analysis. The results showed that high G + C mol% bacteria in hindguts of piglets disappeared after weaning and restored gradually two weeks later. Sequencing analysis indicated that most of these high G + C mol% bacteria blonged to Lactobacillus spp. . Statistical analysis showed that treatment with S1 had no marked effect on diversity index of predominant bacteria from hindguts in piglets. By comparing the bands in DGGE profiles between two groups, a specific band in treatment group was found in profiles from piglets at 14 days of age, sequence matched with that showed 95 % similarity to Clostridium disporicum. At 35 days of age, another specific band appeared in control group, which was identified to be Streptococcus suis (99% ).
ACTA MICROBIOLOGICA SINICA 01/2007; 46(6):961-6.
