A short phylogenetic marker previously used in the reconstruction of the Order Bacillales and the genus Bacillus was assessed here at a lower taxa level: species in the Bacillus cereus group: B. anthracis, B. cereus, ...A short phylogenetic marker previously used in the reconstruction of the Order Bacillales and the genus Bacillus was assessed here at a lower taxa level: species in the Bacillus cereus group: B. anthracis, B. cereus, B. thuringiensis and B. weihenstephanensis. This maker is 220 bp in length. It is a combination of 150 bp at the 3’ end of the 16S rDNA and 70 bp at the 5’ end of the 16S-23S ITS sequence. Three additional Bacillus species, B. halodurans, B. licheniformis and B. subtilis, and Clostridium tetani were included for comparison purposes. A total of eight bacterial species and 12 strains were analyzed. A boot- strapped neighbor-joining tree was inferred from comparative analyses of all allelic sequences of the bacterial species and strains under study. Based on its topology, four major Groups were revealed at the 90% nucleotide sequence identities, Group I to IV. Group I contains all al-leles of the Bacillus cereus group. Group II con-tains all alleles of B. halodurans. Group III con-tains all alleles of B. licheniformis and B. subtilis. Group IV contains all alleles of Clostridium tetani. The 220 bp phylogenetic marker used here could resolve different species from different genera. At the genus level, distant species could be dis-tinguished. Very closely-related species, however, were undistinguishable. Species in the B. cereus group, most notably B. cereus, B. anth- racis and B. thuringiensis, could not be distin- guished. After successfully inferring the phylo- genies of the Order Bacillales and the genus Bacillus, we have met the resolving limit of this short phy-logenetic marker: B. cereus, B. anthracis and B. thuringiensis.展开更多
根据不同种属细菌的16 S rDNA序列两端的保守性区域设计通用引物,提取菌株的基因组DNA,对菌株的16 S rDNA进行了PCR扩增,对扩增到的目标片段进行了测序,将测序结果与NCBI上已知菌种的16 S rDNA序列进行了BLAST对比,并构建了系统进化树...根据不同种属细菌的16 S rDNA序列两端的保守性区域设计通用引物,提取菌株的基因组DNA,对菌株的16 S rDNA进行了PCR扩增,对扩增到的目标片段进行了测序,将测序结果与NCBI上已知菌种的16 S rDNA序列进行了BLAST对比,并构建了系统进化树。结合传统的形态观察及生理生化特性鉴定,16 S rDNA序列分析结果证实芽孢杆菌Pab02为枯草芽孢杆菌,PAS38为蜡样芽孢杆菌。展开更多
Objective:To screen and isolate an eco-friendly,u thermophilic and potent L-asparaginase producing bacterium,with novel immunological properties that may obviates hypersensitivity reactions.Methods:In the present stud...Objective:To screen and isolate an eco-friendly,u thermophilic and potent L-asparaginase producing bacterium,with novel immunological properties that may obviates hypersensitivity reactions.Methods:In the present study baclerial strain isolated for extracellular L-asparaginase production from hotspring,identified by morphological,biochemical and physiological tests followed by t6S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay.Result:The bacterial strain was identified as Bacillus sublilis strain hswx88(GenBank Accession Number JQ237656.1).The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88(23.8 IU/mL)was found to be 1.7 and 14.5 limes higher than the reference organism Pectobacterium carotovorum MTCC 1428(14.2 IU/mL)and Bacillus sp.BCCS 034(1.64 IU/mL).Conclusion:The isolate is eco-friendly and useful to produce bulk quantity of extracellular,thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.展开更多
Background： The chicken gastrointestinal tract contains a diverse microbiota whose composition and structure play important roles in gut functionality. In this study, microbial shifts resulting from feed supplementat...Background： The chicken gastrointestinal tract contains a diverse microbiota whose composition and structure play important roles in gut functionality. In this study, microbial shifts resulting from feed supplementation with Bacillus subtilis CSL2 were evaluated in broilers challenged and unchallenged with Salmonella Gallinarum. To analyse bacterial community composition and functionality, 454 GS-FLX pyrosequencing of 16 S r RNA gene amplicons was performed.Results： The Quantitative Insights into Microbial Ecology（QIIME） pipeline was used to analyse changes in the faecal microbiota over a 24-h period. A total of 718,204 sequences from broiler chickens were recorded and analysed. At the phylum level, Firmicutes, Bacteroidetes, and Proteobacteria were the predominant bacterial taxa. In Salmonellainfected chickens（SC）, Bacteroidetes were more highly abundant compared to control（NC） and Bacillus-treated（BT）chickens. At the genus level, in the NC and BT groups, Lactobacil us was present at high abundance, and the abundance of Turicibacter, unclassified Enterobacteriaceae, and Bacteroides increased in SC broilers. Furthermore, taxon-independent analysis showed that the SC and BT groups were compositional y distinct at the end of the 24-h period. Further analysis of functional properties showed that B. subtilis CSL2 administration increased gut-associated energy supply mechanisms（i.e. carbohydrate transport and metabolism） to maintain a stable microbiota and protect gut integrity.Conclusions： This study demonstrated that S. Gallinarum infection and B. subtilis CSL2 supplementation in the diet of broiler chickens influenced the diversity, composition, and functional diversity of the faecal microbiota. Moreover, the findings offer significant insights to understand potential mechanisms of Salmonel a infection and the mode of action of probiotics in broiler chickens.展开更多
Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluoresce...Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization （FISH） was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LY was 71.7 %. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LY was 74.7% The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.展开更多
The study was sought to enhance the synthesis of thermal stableβ-cyclodextrin glycosyltransferase(β-CGTase)using potato wastewater as a low-cost medium and assess the degree to which it is efficient for industrial p...The study was sought to enhance the synthesis of thermal stableβ-cyclodextrin glycosyltransferase(β-CGTase)using potato wastewater as a low-cost medium and assess the degree to which it is efficient for industrial production ofβ-cyclodextrin(β-CD)from raw potato starch.Thermophilic bacteria producingβ-CGTase was isolated from Saudi Arabia and the promising strain was identified as Bacillus licheniformis using phylogenetic analysis of the 16S rRNA gene.Alginate-encapsulated cultures exhibited twice-fold ofβ-CGTase production more than free cells.Scanning electron microscopy(SEM)of polymeric capsules indicated the potential for a longer shelf-life,which promotes the restoration of activity in bacterial cells across semi-continuous fermentation ofβ-CGTase production for 252 h.The optimal conditions forβ-CGTase synthesis using potato wastewater medium were at 36 h,pH of 8.0,and 50°C with 0.4%potato starch and 0.6%yeast extract as carbon and nitrogen sources,respectively.The purified enzyme showed a specific activity of 63.90 U/mg with a molecular weight of∼84.6 kDa as determined by SDS-PAGE analysis.The high enzyme activity was observed up to 60°C,and complete stability was achieved at 75°C.High levels of activity and stability were shown at pH 8.0,and the pH range from 7.0–10.0,respectively.The enzyme has an appreciable affinity for raw potato starch with a Km of 5.7×10−6 M and a Vmax of 87.71μmoL/mL/min.β-CD production was effective against 25 U/g of raw potato starch.The outcomes demonstrated its feasibility to develop a fermentation process by integrating the cost-effective production ofβ-CGTase having distinctive properties forβ-CD production with ecofriendly utilization of potato wastewater.展开更多
文摘A short phylogenetic marker previously used in the reconstruction of the Order Bacillales and the genus Bacillus was assessed here at a lower taxa level: species in the Bacillus cereus group: B. anthracis, B. cereus, B. thuringiensis and B. weihenstephanensis. This maker is 220 bp in length. It is a combination of 150 bp at the 3’ end of the 16S rDNA and 70 bp at the 5’ end of the 16S-23S ITS sequence. Three additional Bacillus species, B. halodurans, B. licheniformis and B. subtilis, and Clostridium tetani were included for comparison purposes. A total of eight bacterial species and 12 strains were analyzed. A boot- strapped neighbor-joining tree was inferred from comparative analyses of all allelic sequences of the bacterial species and strains under study. Based on its topology, four major Groups were revealed at the 90% nucleotide sequence identities, Group I to IV. Group I contains all al-leles of the Bacillus cereus group. Group II con-tains all alleles of B. halodurans. Group III con-tains all alleles of B. licheniformis and B. subtilis. Group IV contains all alleles of Clostridium tetani. The 220 bp phylogenetic marker used here could resolve different species from different genera. At the genus level, distant species could be dis-tinguished. Very closely-related species, however, were undistinguishable. Species in the B. cereus group, most notably B. cereus, B. anth- racis and B. thuringiensis, could not be distin- guished. After successfully inferring the phylo- genies of the Order Bacillales and the genus Bacillus, we have met the resolving limit of this short phy-logenetic marker: B. cereus, B. anthracis and B. thuringiensis.
文摘根据不同种属细菌的16 S rDNA序列两端的保守性区域设计通用引物,提取菌株的基因组DNA,对菌株的16 S rDNA进行了PCR扩增,对扩增到的目标片段进行了测序,将测序结果与NCBI上已知菌种的16 S rDNA序列进行了BLAST对比,并构建了系统进化树。结合传统的形态观察及生理生化特性鉴定,16 S rDNA序列分析结果证实芽孢杆菌Pab02为枯草芽孢杆菌,PAS38为蜡样芽孢杆菌。
基金Support by the All India Council for Technical Education.New Delhi,India through"Research Promotion Scheme"(AICTE Letter-F.No.8023/RID/RPS-17/(POLICYIV)/(GOVT.)/2011-12.Dt:09/07/2012,Veh.No.:340)
文摘Objective:To screen and isolate an eco-friendly,u thermophilic and potent L-asparaginase producing bacterium,with novel immunological properties that may obviates hypersensitivity reactions.Methods:In the present study baclerial strain isolated for extracellular L-asparaginase production from hotspring,identified by morphological,biochemical and physiological tests followed by t6S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay.Result:The bacterial strain was identified as Bacillus sublilis strain hswx88(GenBank Accession Number JQ237656.1).The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88(23.8 IU/mL)was found to be 1.7 and 14.5 limes higher than the reference organism Pectobacterium carotovorum MTCC 1428(14.2 IU/mL)and Bacillus sp.BCCS 034(1.64 IU/mL).Conclusion:The isolate is eco-friendly and useful to produce bulk quantity of extracellular,thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.
基金supported by a grant from the Next-Generation Bio Green 21Program(PJ01115903),Rural Development Administration,Republic of Korea
文摘Background： The chicken gastrointestinal tract contains a diverse microbiota whose composition and structure play important roles in gut functionality. In this study, microbial shifts resulting from feed supplementation with Bacillus subtilis CSL2 were evaluated in broilers challenged and unchallenged with Salmonella Gallinarum. To analyse bacterial community composition and functionality, 454 GS-FLX pyrosequencing of 16 S r RNA gene amplicons was performed.Results： The Quantitative Insights into Microbial Ecology（QIIME） pipeline was used to analyse changes in the faecal microbiota over a 24-h period. A total of 718,204 sequences from broiler chickens were recorded and analysed. At the phylum level, Firmicutes, Bacteroidetes, and Proteobacteria were the predominant bacterial taxa. In Salmonellainfected chickens（SC）, Bacteroidetes were more highly abundant compared to control（NC） and Bacillus-treated（BT）chickens. At the genus level, in the NC and BT groups, Lactobacil us was present at high abundance, and the abundance of Turicibacter, unclassified Enterobacteriaceae, and Bacteroides increased in SC broilers. Furthermore, taxon-independent analysis showed that the SC and BT groups were compositional y distinct at the end of the 24-h period. Further analysis of functional properties showed that B. subtilis CSL2 administration increased gut-associated energy supply mechanisms（i.e. carbohydrate transport and metabolism） to maintain a stable microbiota and protect gut integrity.Conclusions： This study demonstrated that S. Gallinarum infection and B. subtilis CSL2 supplementation in the diet of broiler chickens influenced the diversity, composition, and functional diversity of the faecal microbiota. Moreover, the findings offer significant insights to understand potential mechanisms of Salmonel a infection and the mode of action of probiotics in broiler chickens.
基金supported by the National Natural Science Foundation of China (No. 20176027).
文摘Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization （FISH） was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LY was 71.7 %. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LY was 74.7% The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.
基金Deanship of Scientific Research at King Khalid University through research groups program,Grant No.R.G.P.1/241/41.
文摘The study was sought to enhance the synthesis of thermal stableβ-cyclodextrin glycosyltransferase(β-CGTase)using potato wastewater as a low-cost medium and assess the degree to which it is efficient for industrial production ofβ-cyclodextrin(β-CD)from raw potato starch.Thermophilic bacteria producingβ-CGTase was isolated from Saudi Arabia and the promising strain was identified as Bacillus licheniformis using phylogenetic analysis of the 16S rRNA gene.Alginate-encapsulated cultures exhibited twice-fold ofβ-CGTase production more than free cells.Scanning electron microscopy(SEM)of polymeric capsules indicated the potential for a longer shelf-life,which promotes the restoration of activity in bacterial cells across semi-continuous fermentation ofβ-CGTase production for 252 h.The optimal conditions forβ-CGTase synthesis using potato wastewater medium were at 36 h,pH of 8.0,and 50°C with 0.4%potato starch and 0.6%yeast extract as carbon and nitrogen sources,respectively.The purified enzyme showed a specific activity of 63.90 U/mg with a molecular weight of∼84.6 kDa as determined by SDS-PAGE analysis.The high enzyme activity was observed up to 60°C,and complete stability was achieved at 75°C.High levels of activity and stability were shown at pH 8.0,and the pH range from 7.0–10.0,respectively.The enzyme has an appreciable affinity for raw potato starch with a Km of 5.7×10−6 M and a Vmax of 87.71μmoL/mL/min.β-CD production was effective against 25 U/g of raw potato starch.The outcomes demonstrated its feasibility to develop a fermentation process by integrating the cost-effective production ofβ-CGTase having distinctive properties forβ-CD production with ecofriendly utilization of potato wastewater.
