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84 Chapter IV: Results by neighbor-joining method using MEGA 5 package (Tamura et al., 2007). Gaps were treated as missing data. Only unambiguous alignments were used in phylogenetic analyses. The complete 16S rRNA sequence analysis revealed that bacterial isolates had 77% to 100% similarity with the sequences of NCBI database. The isolate LKH-3 was identified as Lactobacillus helveticus as it showed 100% resemblance with it. On the basis of phylogenetic data, isolate Lamec-29 had close resemblance with members of the genus Lactobacillus. The percentage similarities were calculated between Lamec-29 and Lactobacillus fermentum and found to be similar approximate 86%. The isolate LAM-1 had shown maximum similarity with Lactobacillus casei which is known to probiotic bacterium. The isolate LKH-2 showed 97% similarity with two different Lactobacillus species, Lactobacillus delbrueckii subsp. lactis and Lactobacillus delbrueckii subsp. bulgaricus. The percentage differences calculated between isolate LAM-2 and three different species of Lactobacillus casei, viz., Lactobacillus casei AB605428, Lactobacillus casei HQ117896 and Lactobacillus casei JN560922 suggest that this may represent a new species of genus Lactobacillus. The isolate LKH-3 showed 96% homology with two different Lactobacillus species, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus delbrueckii subsp. The phylogenetic tree identified this isolate as Lactobacillus spp. It is interesting that the sources of all the isolates of NCBI database which were used to construct phylogenetic tree were probiotic bacteria. It matches well with the fact that isolates of present study were also isolated from traditional food and human gut and posess probiotic attributes. It is noteworthy that isolates Lamec-29, LKH-2, LKH-3 and LAM-2 16S rRNA did not show high identity with sequences in the database. The sequences that share an identity below 98% are usually considered to be part of the same genus (Sadowsky et al., 1996). On this basis, these isolates described here probably represent a new member of a known genus (Lactobacillus), which
85 Chapter IV: Results has probiotic attributes. The 16S rRNA gene sequences of all six bacterial isolates determined in this study were deposited in the GenBank of NCBI data library under accession numbers JN620211, JN618457, JN620214, JN620213, JN620212 and JN620215 for LAM-1, LAM-2, LKH-2, LKH-3, LKH-5, LS-2 and Lamec-29 respectively (Table 4.7, 4.8, 4.9, 4.10, 4.11and 4.12). 84 42 0.01 98 85 76 52 48 100 39 34 35 65 77 79 100 LKH-5 (JN620212) Lactobacillus helveticus EU377824 Lactobacillus helveticus AB680751 Lactobacillus helveticus AB334764 Lactobacillus fermentum JN039355 Lactobacillus fermentum EU407607 Lamec-29 (JN620215) Lactobacillus fermentum EU419596 Lactobacillus delbrueckii subsp. lactis JQ580992 100 99 Lactobacillus delbrueckii subsp. lactis AB681888 100 77 69 LKH-2(JN620214) Lactobacillus delbrueckii subsp. bulgaricus HQ293115 Lactobacillus delbrueckii subsp. bulgaricus EF100616 LKH-3 (JN620213) Lactobacillus delbrueckii subsp. bulgaricus AY138279 Lactobacillus casei AB605428 LAM-2 (JN620211) Lactobacillus casei HQ286593 LAM-1 (JN618457) Lactobacillus casei HQ117896 Lactobacillus casei JN560923 Lactobacillus casei JN560924 Lactobacillus casei JN560922 Bacillus subtilis AB374521 Fig. 4.3 Neighbor-joining tree based on bacterial 16S rRNA sequence data from different isolates of current study along with sequences available in GenBank database. Numerical values indicate bootstrap percentile from 1000 replicates. Bacillus subtilis was used as outgroup taxa.
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Characterization of Lactobacilli is
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their encouragement, insightful com
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5. Mukesh Kumar and Abhijit Ganguli
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2.3.2.9 Cholesterol lowering 2.3.2.
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3.12.4 Measurement of intracellular
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List of Abbreviations ABTS 2,2-azin
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LIST OF TABLES Tables Page No. Tabl
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Chapter 1 INTRODUCTION
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2 Chapter I: Introduction Recent re
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4 Chapter I: Introduction Gram-posi
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6 Chapter I: Introduction food-born
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8 Chapter I: Introduction beneficia
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Chapter I1 REVIEW OF LITERATURE
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Chapter II Review of Literature nee
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13 Chapter II Review of Literature
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Page 67 and 68: Chapter II1 MATERIALS & METHODS
Page 69 and 70: 50 Chapter III Material and methods
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Page 139 and 140: Chapter V DISCUSSION
Page 141 and 142: 120 Chapter V Discussion al., 2003)
Page 143 and 144: 122 Chapter V Discussion Lactobacil
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134 Chapter V Discussion would prev
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136 Chapter V Discussion gave maxim
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138 Chapter V Discussion for 90 min
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140 Chapter V Discussion To test th
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CONCLUSIONS The main objective of t
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145 Chapter V: References Aasen, I.
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147 Chapter V: References Bjorksten
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149 Chapter V: References negative,
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151 Chapter V: References Delves-Br
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153 Chapter V: References Franz, C.
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155 Chapter V: References Gordon, D
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157 Chapter V: References Isolauri,
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159 Chapter V: References Kolenbran
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161 Chapter V: References Marekova,
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163 Chapter V: References Nagy, A.,
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165 Chapter V: References Bifidobac
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167 Chapter V: References Schilling
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169 Chapter V: References Teuber, M
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171 Chapter V: References Yoon, Y.C
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Rogosa agar Ingredients Quantity (g
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Denaturing solution. guanidinium is
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Appendix II Fig. 3.1. Standard curv
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