scholarly journals Gammaproteobacteria as essential primary symbionts in the striped shield bug, Graphosoma Lineatum (Hemiptera: Pentatomidae)

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Naeime Karamipour ◽  
Yaghoub Fathipour ◽  
Mohammad Mehrabadi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Adult Stage ◽  
Phylogenetic Analyses ◽  
Light And Electron Microscopy ◽  
Pcr Analysis ◽  
Evolutionary Analysis ◽  
Bacterial Cells ◽  
Stink Bug ◽  
Surface Sterilization

Abstract Many members of suborder Heteroptra harbor heritable symbiotic bacteria. Here we characterize the gut symbiotic bacterium in Graphosoma lineatum (Hemiptera: Pentatomidae) by using molecular phylogeny, real-time PCR analysis as well as light and electron microscopy observations. The microscopy observations revealed the presence of a large number of rod-shaped bacterial cells in the crypts. A very high prevalence (98 to 100%) of the symbiont infection was found in the insect populations that strongly supports an intimate association between these two organisms. Real-time PCR analysis also showed that the Gammaproteobacteria dominated the crypts. The sequences of 16sr RNA and groEL genes of symbiont showed high levels of similarity (93 to 95%) to Pantoea agglomeranse and Erwinia herbicola Gammaproteobacteria. Phylogenetic analyses placed G. lineatum symbiont in a well-defined branch, divergent from other stink bug bacterial symbionts. Co-evolutionary analysis showed lack of host-symbiont phylogenetic congruence. Surface sterilization of eggs resulted in increased pre-adult stage in the offspring (aposymbionts) in comparison to the normal. Also, fecundity, longevity, and adult stage were significantly decreased in the aposymbionts. Therefore, it seems that the symbiont might play a vital function in the host biology, in which host optimal development depends on the symbiont.

Habitat visualization, acquisition features and necessity of the gammaproteobacterial symbiont of pistachio stink Bug, Acrosternum heegeri (Hem.: Pentatomidae)

2019 ◽  
Vol 110 (1) ◽  
pp. 22-33 ◽  
Author(s):  
M. Kashkouli ◽  
Y. Fathipour ◽  
M. Mehrabadi
Keyword(s):  
Electron Microscopy ◽  
Real Time ◽  
Real Time Pcr ◽  
Survival Rates ◽  
Adult Survival ◽  
Bacterial Cells ◽  
Stink Bug ◽  
Surface Sterilization ◽  
Posterior Midgut ◽  
Molecular Phylogenetic

AbstractPlant-sucking stinkbugs are especially associated with mutualistic gut bacterial symbionts. Here, we explored the symbiotic relationship of a pistachio stinkbug, Acrosternum heegeri Fieber by histological, fluorescence in situ hybridization (FISH), real-time PCR and molecular phylogenetic techniques. Furthermore, the effects of the symbiont on the resting/wandering behaviors of the newborn nymphs, pre-adult survival rates, and stage compositions were investigated. Transmission electron microscopy and real-time PCR analyses showed that a rod-shaped gammaproteobacterium was persistently located within the posterior midgut crypts. Molecular phylogenetic and FISH techniques strongly suggested that this symbiont should be placed in the genus Pantoea of the Enterobacteriales. Scanning electron microscopy confirmed the presence of the bacterial cells on the egg surface which the surface sterilization of the eggs resulted in the successful removal of the symbiont from the eggs. Symbiotic and aposymbiotic A. heegeri showed no significant differences in the wandering behaviors of the first nymphal stages, while the symbiont-free insects suffered retarded growth and lower survivability. Together, the results highlight the habitat and acquisition features of Pantoea symbiont and its contribution in A. heegeri biology that might help us for better pest management in the future.

scholarly journals Real-Time PCR Quantitation of Clostridia in Feces of Autistic Children

2004 ◽  
Vol 70 (11) ◽  
pp. 6459-6465 ◽  
Author(s):  
Yuli Song ◽  
Chengxu Liu ◽  
Sydney M. Finegold
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Late Onset ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Pcr Analysis ◽  
Bacterial Cells ◽  
Autistic Children ◽  
Bacterial Strains ◽  
And Control

ABSTRACT Based on the hypothesis that intestinal clostridia play a role in late-onset autism, we have been characterizing clostridia from stools of autistic and control children. We applied the TaqMan real-time PCR procedure to detect and quantitate three Clostridium clusters and one Clostridium species, C. bolteae, in stool specimens. Group- and species-specific primers targeting the 16S rRNA genes were designed, and specificity of the primers was confirmed with DNA from related bacterial strains. In this procedure, a linear relationship exists between the threshold cycle (CT ) fluorescence value and the number of bacterial cells (CFU). The assay showed high sensitivity: as few as 2 cells of members of cluster I, 6 cells of cluster XI, 4 cells of cluster XIVab, and 0.6 cell of C. bolteae could be detected per PCR. Analysis of the real-time PCR data indicated that the cell count differences between autistic and control children for C. bolteae and the following Clostridium groups were statistically significant: mean counts of C. bolteae and clusters I and XI in autistic children were 46-fold (P = 0.01), 9.0-fold (P = 0.014), and 3.5-fold (P = 0.004) greater than those in control children, respectively, but not for cluster XIVab (2.6 � 108 CFU/g in autistic children and 4.8 � 108 CFU/g in controls; respectively). More subjects need to be studied. The assay is a rapid and reliable method, and it should have great potential for quantitation of other bacteria in the intestinal tract.

Broad Range Evaluation of the Matrix Solubilization (Matrix Lysis) Strategy for Direct Enumeration of Foodborne Pathogens by Nucleic Acids Technologies

2009 ◽  
Vol 72 (6) ◽  
pp. 1225-1233 ◽  
Author(s):  
EVA MAYRL ◽  
BARBARA ROEDER ◽  
PATRICK MESTER ◽  
MARTIN WAGNER ◽  
PETER ROSSMANITH
Keyword(s):  
Salmonella Typhimurium ◽  
Real Time ◽  
Real Time Pcr ◽  
Sodium Dodecyl ◽  
Ice Cream ◽  
Model Organisms ◽  
Pcr Analysis ◽  
Relative Standard Error ◽  
Bacterial Cells ◽  
Relative Standard

A previously published rapid (<5 h) proof-of-concept protocol for the concentration of the foodborne pathogen Listeria monocytogenes from milk, based on the solubilization of the food matrix, was further evaluated. The original protocol was modified to detect gram-negative and other gram-positive bacteria and to broaden the range of food matrices by using Lutensol instead of sodium dodecyl sulfate as the main detergent in the buffer. A new protocol using a protease and sucrose buffer was established for the analysis of meat and fish. Matrix lysis was used for dairy products, ice cream, milk, fish, meat, eggs, and blood. Solubilization of the foodstuffs resulted in bacterial pellets of reasonable size for further quantification. Using L. monocytogenes, Staphylococccus aureus, Bacillus cereus, Escherichia coli, and Salmonella Typhimurium as model organisms, microscopic analysis of the remaining bacterial pellets revealed that the recovered bacteria remained physically intact, albeit their viability was compromised. In model experiments using free DNA, the free target DNA was reduced by 5 log units. The compatibility of matrix lysis with subsequent real-time PCR analysis has been demonstrated with salmon, chicken, egg, ice cream, cheese, and blood samples that were artificially contaminated with L. monocytogenes, S. aureus, and Salmonella Typhimurium. These experiments resulted in an average recovery of 48.7% (relative standard error, 83.4%) of the inoculated bacterial cells with the real-time PCR assay. The average detection limit of the method was 7.3 CFU/ml for all examined foodstuffs and bacterial target organisms.

ID: 316 Real-time PCR analysis of BCL2L12, a novel member of BCL2 family of the apoptosis-related genes, in human leukemia.

2006 ◽  
Vol 4 (s1) ◽  
pp. 82-82
Author(s):  
K. Floros ◽  
H. Thomadaki ◽  
S. Pavlovic ◽  
M. Talieri ◽  
M. Colovic ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Analysis ◽  
Human Leukemia ◽  
Bcl2 Family

Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens

2021 ◽  
Vol 70 (9) ◽  
Author(s):  
Berta Fidalgo ◽  
Elisa Rubio ◽  
Victor Pastor ◽  
Marta Parera ◽  
Clara Ballesté-Delpierre ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Enteric Pathogens ◽  
Pcr Analysis ◽  
Culture Methods ◽  
Gastrointestinal Infections ◽  
Content Type ◽  
Link Type ◽  
Microbiological Culture ◽  
Micro Organisms

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis. Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination. Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures. Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013–2018) included stool culture, microscopy and immunochromatography. Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013–2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis. Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.

scholarly journals Real-time PCR analysis of the apoptosis related genes in ATRA treated APL t(15;17) patients

2003 ◽  
Vol 35 (5) ◽  
pp. 454-459 ◽  
Author(s):  
Hakan Savli ◽  
Sema Sirma ◽  
Balint Nagy ◽  
Melih Aktan ◽  
Guncag Dincol ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Analysis
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