By Culture Techniques (by + culture_techniques)

Distribution by Scientific Domains


Selected Abstracts


Antibacterial efficacy of calcium hydroxide intracanal dressing: a systematic review and meta-analysis

INTERNATIONAL ENDODONTIC JOURNAL, Issue 1 2007
C. Sathorn
Abstract Aim, To determine to what extent does calcium hydroxide intracanal medication eliminate bacteria from human root canals, compared with the same canals before medication, as measured by the number of positive cultures, in patients undergoing root canal treatment for apical periodontitis (teeth with an infected root canal system). Methodology, CENTRAL, MEDLINE and EMBASE databases were searched. Reference lists from identified articles were scanned. A forward search was undertaken on the authors of the identified articles. Papers that had cited these articles were also identified through the Science Citation Index to identify potentially relevant subsequent primary research. Review methods, The included studies were pre-/post-test clinical trials comparing the number of positive bacterial cultures from treated canals. Data in those studies were independently extracted. Risk differences of included studies were combined using the generic inverse variance and random effect method. Results, Eight studies were identified and included in the review, covering 257 cases. Sample size varied from 18 to 60 cases; six studies demonstrated a statistically significant difference between pre- and post-medicated canals, whilst two did not. There was considerable heterogeneity among studies. Pooled risk difference was ,21%; 95% CI: ,47% to 6%. The difference between pre- and post-medication was not statistically significant (P = 0.12). Conclusions, Calcium hydroxide has limited effectiveness in eliminating bacteria from human root canal when assessed by culture techniques. [source]


The persistence of bifidobacteria populations in a river measured by molecular and culture techniques

JOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2009
X. Bonjoch
Abstract Aims:, To determine relative to faecal coliforms (FC) and sulfite-reducing clostridia (SRC), the environmental persistence of natural populations of Bifidobacterium spp. enumerated by culturing and quantitative polymerase chain reaction (q-PCR). Methods and Results:, Dialysis tubing containing river supplemented with overnight cultures of Bifidobacterium adolescentis (BA) and Bifidobacterium dentium (BD) or urban wastewater were suspended in a river for up to 10 days. At intervals, the contents of each dialysis tube were assayed using q-PCR assays for BA and BD, and selective culture media for FC, SRC, total bifidobacteria (TB), sorbitol-fermenting bifidobacteria (SFB) and cultivable BA. Mean summer T90 values were 251 h for SRC, 92 h for FC, 48 h for BA and BD by q-PCR, and 9 h for TB. Conclusions:,Bifidobacterium spp. was the population with the lowest persistence, showing seasonal differences in T90 when measured by culture techniques or by q-PCR. This difference in relative persistence is because of a longer persistence of molecular targets than cultivable cells. Significance and Impact of the Study:, The persistence of a viable bifidobacteria cells is shorter, but the longest persistence of molecular targets. This factor could be used for origin the faecal pollution in water for the development of microbial source tracking (MST). [source]


Influence of genetic background on transformation and expression of Green Fluorescent Protein in Actinobacillus actinomycetemcomitans

MOLECULAR ORAL MICROBIOLOGY, Issue 5 2005
W. Teughels
Background/aims:, The development of an electro-transformation system and the construction of shuttle plasmids for Actinobacillus actinomycetemcomitans have enhanced the molecular analysis of virulence factors. However, inefficient transformation is frequently encountered. This study investigated the efficiency of electro-transformation and expression of Green Fluorescent Protein (GFP) in 12 different A. actinomycetemcomitans strains. The influence of the plasmid vector, serotype, and phenotype were the major factors taken into consideration. Material and methods:, Twelve serotyped A. actinomycetemcomitans strains were independently electro-transformed with two different Escherichia coli,A. actinomycetemcomitans shuttle plasmids (pVT1303 and pVT1304), both containing an identical ltx-GFPmut2 gene construct but a different backbone (pDMG4 and pPK1, respectively). The transformation efficiency, transformation frequency, and electro-transformation survival rate were determined by culture techniques. GFP expression was observed at the colony level by fluorescence microscopy. Results:, All strains could be transformed with both plasmids. However, major differences were observed for the transformation efficiency, transformation frequency, and electro-transformation survival rate between strains. The data demonstrated that plasmid vector, serotype, and phenotype are key players for obtaining a successful transformation. An inverted relationship between the electro-transformation survival rate and tranformation frequency was also observed. GFP expression was also influenced by phenotype, serotype and plasmid vector. Conclusions:, The serotype of A. actinomycetemcomitans has an important influence on its survival after electro-transformation and on transformation frequency. The expression of GFP is strain and plasmid vector dependent. [source]


Comparison of Growth and Recombinant Protein Expression in Two Different Insect Cell Lines in Attached and Suspension Culture

BIOTECHNOLOGY PROGRESS, Issue 4 2001
R. A. Taticek
Culture conditions required for obtaining maximum recombinant protein concentrations from two cell lines, Spodoptera frugiperda (IPL,-Sf21-AE) and Trichoplusia ni (Tn 5,-1,4), were determined in this work. Conditions studied include mode of culture (suspended vs attached), agitation rates, inoculum sizes, cell concentration at the time of infection, and various serum-free media (SFM). Results were compared with the performance of attached cultures in TnM-FH with 10% fetal bovine serum. Growth rates in the different culture media tested were similar, but the cell numbers achieved (i.e., yield) improved 2 to 2.7-fold in SFM over cultures in TnM-FH. Agitation rates of 150,160 rpm were necessary for maximum growth of suspended Tn 5,-1,4 cells compared to 125,150 rpm for Sf-21 cells. An inoculum size of 5 × 105 cells/mL gave good growth rates and optimum biomass yields for both cell lines. Cultures of both cell lines were infected with viruses encoding for ,-galactosidase or human secreted alkaline phosphatase (seAP). Protein expression in TnM-FH in attached culture showed that Tn 5,-1,4 cells are 2,4.5 times more productive on a per cell basis than Sf-21 cells grown under similar conditions. Production of ,-galactosidase in Sf-21 cells increased 50% in suspension cultures with SFM compared to attached cultures in TnM-FH, but seAP expression was essentially unchanged by culture techniques. The Tn 5,-1,4 cells produced 2.6,4.4 and 2.7,3 times more ,-galactosidase and seAP, respectively, in SFM in suspension compared to Sf-21 cells. EX-CELL 401 and Sf900-II were formulated as optimized SFM for Sf cell lines. However, in Sf-21 cultures EX-CELL 400 performed better than the other two media, as it increased the ,-galactosidase yield up to 25%. Surprisingly, EX-CELL 401 was the best medium for the production of ,-galactosidase by Tn 5,-1,4 cells, resulting in 25% and 69% higher volumetric and specific yields, respectively, compared to EX-CELL 405 which was formulated for this specific cell line. These results show that even when culture media are designed for maximal growth of a specific cell line, other media may provide the best conditions for protein production. [source]