scholarly journals Detection of Gram-negative Bacteria in Broad-range PCR Amplifying 16S rRNA Gene with Semi-nested Primers and Its Application in Market Milk

2005 ◽  
Vol 47 (3) ◽  
pp. 465-474
2013 ◽  
Vol 18 (1) ◽  
pp. 67
Author(s):  
Fahrizal Hazra ◽  
Etty Pratiwi

The objectives of the research were: (i)  to isolate and characterize of phosphate solubilizing bacteria (PSB) and (ii) to identify PSB based on molecular amplification of 16S rRNA gene.  Soil samples were collected from rhizosphere in Bogor, West Nusa Tenggara, and East Nusa Tenggara.  Several stages in this research were: (i) isolation PSB in Pikovskaya agar, (ii) morphological and biochemical characterization of PSB, (iii) measurement of  phosphatase enzymes, and (iv) measurement of secreting indole acetic acid phytohormone.   As many as 29 isolates of PSB have been collected and three isolates of them, namely: P 3.5 (East Nusa Tenggara), P 6.2 (West Nusa Tenggara), and P 10.1 (Citeureup, West Java) were chosen for further study.  There were many characteristics of isolate P 10.1: (i) it had capable to solubilize P with the value of highest solubilization index (1.80), (ii) it had the highest phosphatase enzyme (120.40 mg kg-1), and (iii) it had the highest pH decrease at each observation for six days.  Isolates P 3.5 and P 10.1 were the Gram-negative bacteria with coccus shapes and isolate P 6.2 was a Gram-negative bacteria with bacillus shape.  Deoxiribonucleat Acid (DNA) amplification of these bacteria employing 16S rRNA primers generated the 1,300bp-PCR product.  The results of the analysis of 16S rRNA gene sequences showed that isolates P 3.5 and P 10.1 has 98% similarity with Gluconacetobacter sp. strains Rg1-MS-CO and isolate P 6.2 has 97% similarity with Enterobacter sp. pp9c strains.Keywords: 16S rRNA, indole acetic acid, isolation, phosphatase enzymes, phosphate solubilizing bacteria[How to Cite : Hazra F and E Pratiwi. 2013. Isolation, Characterization, and Molecular Identification of Phosphate Solubilizing Bacteria from Several Tropical Soils. J Trop Soils, 18 (1): 67-74. doi: 10.5400/jts.2013.18.1.67][Permalink/DOI: www.dx.doi.org/10.5400/jts.2013.18.1.67]


2011 ◽  
Vol 60 (9) ◽  
pp. 1281-1286 ◽  
Author(s):  
Jade L. L. Teng ◽  
Ming-Yiu Yeung ◽  
Geoffrey Yue ◽  
Rex K. H. Au-Yeung ◽  
Eugene Y. H. Yeung ◽  
...  

2015 ◽  
Vol 19 (1) ◽  
pp. 64
Author(s):  
Sri Darmawati ◽  
Langkah Sembiring ◽  
Widya Asmara ◽  
Wayan T. Artama ◽  
Masashi Kawaichi

The purpose of this study was to analyze the phylogenetic relationship of Gram negative bacteria (3strains of Salmonella typhi, 1 strain of Escherichia coli, 1 strain of Serratia marcescens, and 3 strains of Enterobactercloacae) of Enterobacteriaceae family in positive Widal blood cultures based on 16S rRNA gene sequences. Theresults respectively showed that each two 16S rRNA gene clones of Serratia marcescens KD 08.4 had a closerelationship with 16S rRNA gene of Serrratia marcescens ATCC 13880 (similarity: 99.53-99.8%), Eschericia coliBA 30.1 with Eschericia coli ATCC 11775T (similarity: 99.38-99.67%), Salmonella typhi BA 07.4, Salmonella typhiKD 30.4, and Salmonella typhi SA 02.2 with Salmonella typhi ATCC 19430T (similarity: 99.4-100%) as well as theisolates of Enterobacter cloacae SA 02.1, Enterobacter cloacae BA 45.4.1, one 16S rRNA gene clone of Enterobactercloacae TG 03.5 with Enterobacter cloacae ATCC 23373 (similarity: 99.0-99.87%).


2010 ◽  
Vol 76 (19) ◽  
pp. 6689-6691 ◽  
Author(s):  
W. Vahjen ◽  
R. Pieper ◽  
J. Zentek

ABSTRACT Feeding high levels of zinc oxide to piglets significantly increased the relative abundance of ileal Weissella spp., Leuconostoc spp., and Streptococcus spp., reduced the occurrence of Sarcina spp. and Neisseria spp., and led to numerical increases of all Gram-negative facultative anaerobic genera. High dietary zinc oxide intake has a major impact on the porcine ileal bacterial composition.


2004 ◽  
Vol 54 (6) ◽  
pp. 2223-2230 ◽  
Author(s):  
Linxian Ding ◽  
Akira Yokota

Two strains of curved bacteria, 7-1T and 7-2T, isolated from well water, were phylogenetically examined to determine their taxonomic position. Strain 7-1T is a Gram-negative, slightly curved rod. Analysis of the 16S rRNA gene sequence showed that strain 7-1T formed a cluster with [Aquaspirillum] delicatum and [Pseudomonas] lanceolata. It has some similar characteristics to [A.] delicatum and [P.] lanceolata, but has sufficient distance to separate it from other genera. DNA–DNA hybridization analysis, as well as chemotaxonomic and morphological studies, demonstrated that strain 7-1T, [A.] delicatum and [P.] lanceolata belong to a new genus, Curvibacter gen. nov. Strain 7-1T (=IAM 15033T=ATCC BAA-807T) is classified as the type strain of Curvibacter gracilis gen. nov., sp. nov., and [A.] delicatum and [P.] lanceolata are classified as Curvibacter delicatus comb. nov. and Curvibacter lanceolatus comb. nov., respectively. Strain 7-2T is a Gram-negative spirillum. Phylogenetic study based on the 16S rRNA gene sequences showed that it formed a cluster with the members of the genus Herbaspirillum, [Pseudomonas] huttiensis and [Aquaspirillum] autotrophicum. The classification is therefore proposed of strain 7-2T (=IAM 15032T=ATCC BAA-806T) as the type strain of Herbaspirillum putei sp. nov., and [P.] huttiensis and [A.] autotrophicum are transferred to the genus Herbaspirillum as Herbaspirillum huttiense comb. nov. and Herbaspirillum autotrophicum comb. nov., respectively.


2010 ◽  
Vol 59 (11) ◽  
pp. 1293-1302 ◽  
Author(s):  
Mitsuo Sakamoto ◽  
Moriya Ohkuma

The hsp60 gene sequences were determined for 121 strains of Gram-negative anaerobic rods, including the genera Bacteroides, Barnesiella, Butyricimonas, Odoribacter, Parabacteroides, Paraprevotella, Porphyromonas, Prevotella and Tannerella. The mean pairwise hsp60 gene sequence similarity (73.8–97.1 %) between species in each genus, except for the genus Tannerella that comprises one species, was significantly less than that of the 16S rRNA gene sequence (88.3–96.3 %). Only pairwise hsp60 gene sequence similarity (97.1 %) of the genus Paraprevotella was higher than that of the 16S rRNA gene sequence (93.8 %). Each genus formed a distinct clade in the phylogenetic analysis of the hsp60 gene sequence as well as the 16S rRNA gene sequence. The phylogenetic analysis indicated a higher evolutionary rate for the hsp60 gene sequence than the 16S rRNA gene sequence, especially in the genera Porphyromonas and Prevotella. This study suggests that the hsp60 gene is a useful alternative phylogenetic marker for the identification and classification of a broad range of Gram-negative anaerobic rods.


2010 ◽  
Vol 60 (6) ◽  
pp. 1271-1279 ◽  
Author(s):  
Hélène Marchandin ◽  
Corinne Teyssier ◽  
Josiane Campos ◽  
Hélène Jean-Pierre ◽  
Frédéric Roger ◽  
...  

Three strains of a hitherto unknown, Gram-negative, tiny, anaerobic coccus were collected from human clinical samples originating from skin and soft tissues. The three isolates displayed at least 99.9 % identity in their 16S rRNA gene sequences and more than 99.8 % identity in their dnaK gene sequences. The isolates were affiliated to the family Veillonellaceae, the coccobacillus Dialister micraerophilus being the most closely related species, but there was no more than 91.1 % identity in the 16S rRNA gene sequence between this species and the three isolates. Phylogeny based on the 16S rRNA gene confirmed that the three strains represent a novel and robust lineage within the current family Veillonellaceae. A similar genomic structure was demonstrated for the three isolates by PFGE-based analysis. Morphology and metabolic end products, as well as genotypic and phylogenetic data supported the proposal of the novel genus Negativicoccus gen. nov., with the novel species Negativicoccus succinicivorans sp. nov. [type strain ADV 07/08/06-B-1388T (=AIP 149.07T=CIP 109806T=DSM 21255T=CCUG 56017T) as type species]. Phylogenetic analyses based on the 16S rRNA gene sequences of members of the phylum Firmicutes and other phyla indicated that the family Veillonellaceae forms a robust lineage clearly separated from those of the classes ‘Bacilli’, ‘Clostridia’, Thermolithobacteria and ‘Erysipelotrichi’ in the phylum Firmicutes. Therefore, we propose that this family is a class-level taxon in the phylum Firmicutes, for which the name Negativicutes classis nov. is proposed, based on the Gram-negative type of cell wall of its members, with the type order Selenomonadales ord. nov. In this order, a novel family, Acidaminococcaceae fam. nov., is proposed and description of the family Veillonellaceae is emended.


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