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Isogenic Strains (isogenic + strain)
Selected AbstractsPossible role of the adhesin ace and collagen adherence in conveying resistance to disinfectants on Enterococcus faecalisMOLECULAR ORAL MICROBIOLOGY, Issue 6 2008G. Kayaoglu Introduction:, This study aimed to evaluate whether the presence of the ace gene and Ace-mediated binding to collagen confers on Enterococcus faecalis resistance against common endodontic disinfectants. Methods:, Isogenic strains of E. faecalis: OG1RF (wild-type) and TX5256 (ace insertion mutant of OG1RF) were grown in brain,heart infusion broth at 46°C overnight. Standardized bacterial suspensions were pretreated for 1 h either with acid-soluble collagen or acidified phosphate-buffered saline (ac-PBS). Bacteria were challenged with chlorhexidine digluconate (CHX), iodine potassium-iodide (IKI), sodium hypochlorite (NaOCl), and calcium hydroxide [Ca(OH)2]. Samples were removed at 1, 3, and 6 h, and cultured on Todd,Hewitt agar plates. Colonies were counted, the absolute values were log transformed, and the data were statistically analyzed using Fisher's least significant differences test and t -test. Results:, OG1RF was more resistant than TX5256 to IKI, NaOCl, and Ca(OH)2 (P < 0.05). Collagen-exposed OG1RF was more resistant than the ac-PBS-pretreated OG1RF against CHX at 3 h and against IKI at 1 h (P < 0.05); no significant difference was found against NaOCl. As expected, the ace mutant strain, TX5256, pretreated with collagen or ac-PBS did not differ significantly in viability when challenged with CHX, IKI, and NaOCl. An unexpected result was found for Ca(OH)2: collagen-pretreated OG1RF and TX5256 were both more susceptible than ac-PBS-pretreated OG1RF and TX5256, respectively (P < 0.05). Conclusion:, The presence of the ace gene confers resistance against IKI, NaOCl, and Ca(OH)2 on E. faecalis. Exposure to collagen makes the wild-type bacterium more resistant against CHX and IKI; however, exposure to collagen apparently decreases resistance to Ca(OH)2. [source] Interference with the quorum sensing systems in a Vibrio harveyi strain alters the growth rate of gnotobiotically cultured rotifer Brachionus plicatilisJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2007N.T.N. Tinh Abstract Aims:, To evaluate the effect of Vibrio harveyi strains on the growth rate of the gnotobiotically cultured rotifer Brachionus plicatilis, and to establish whether quorum sensing is involved in the observed phenomena. Methods and Results:, Gnotobiotic B. plicatilis sensu strictu, obtained by hatching glutaraldehyde-treated amictic eggs, were used as test organisms. Challenge tests were performed with 11 V. harveyi strains and different quorum sensing mutants derived from the V. harveyi BB120 strain. Brominated furanone [(5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone] as a quorum sensing inhibitor was tested in Brachionus challenge tests. Some V. harveyi strains, such as strain BB120, had a significantly negative effect on the Brachionus growth rate. In the challenge test with MM77, an isogenic strain of BB120 in which the two autoinducers (HAI-1 and AI-2) are both inactivated, no negative effect was observed. The effect of single mutants was the same as that observed in the BB120 strain. This indicates that both systems are responsible for the growth-retarding (GR) effect of the BB120 strain towards Brachionus. Moreover, the addition of an exogenous source of HAI-1 or AI-2 could restore the GR effect in the HAI-1 and AI-2 nonproducing mutant MM77. The addition of brominated furanone at a concentration of 2·5 mg l,1 could neutralize the GR effect of some strains such as BB120 and VH-014. Conclusions:, Two quorum sensing systems in V. harveyi strain BB120 (namely HAI-1 and AI-2-mediated) are necessary for its GR effect on B. plicatilis. With some other V. harveyi strains, however, growth inhibition towards Brachionus does not seem to be related to quorum sensing. Significance and Impact of the Study:, Interference with the quorum sensing system might help to counteract the GR effect of some V. harveyi strains on Brachionus. However, further studies are needed to demonstrate the positive effect of halogenated furanone in nongnotobiotic Brachionus cultures and eventually, in other segments of the aquaculture industry. [source] Efficacy of various pyrethroid structures against a highly metabolically resistant isogenic strain of Helicoverpa armigera (Lepidoptera: Noctuidae) from ChinaPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 10 2007Jianguo Tan Abstract BACKGROUND: Resistance to pyrethroids and other types of insecticides in Helicoverpa armigera (Hübner) has been documented in many countries. The isolation of specific resistance mechanisms in isogenic strains is an optimal approach to investigate cross-resistance pattern, and to validate resistance breaking pyrethroids. In this study an isogenic metabolic resistance CMR strain was successfully isolated from a field pyrethroid-resistant population of H. armigera. With this strain, cross-resistance among 19 pyrethroid insecticides with varying chemical structures was analysed. RESULTS: Resistance to pyrethroids in the CMR strain was likely to be due to enhanced oxidative metabolism. The most significant cross-resistance in the CMR strain was between pyrethroids such as fenvalerate, tau-fluvalinate and flumethrin characterised by having both phenoxybenzyl and aromatic acid moieties. Substitution of the phenoxybenzyl group with a polyfluorobenzyl group, as in tefluthrin, benfluthrin and transfluthrin, overcame most of this resistance. CONCLUSION: The findings in this study support the assertion that it is possible to find pyrethroids that are active against resistant populations. Such pyrethroids could be considered as possible partners or resistance breaking pyrethroids in a pyrethroid resistance management programme for H. armigera in China and in other Asian countries where the oxidative metabolism resistance is a dominant mechanism. Copyright © 2007 Society of Chemical Industry [source] Developing live Shigella vaccines using , Red recombineeringFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 3 2006Ryan T. Ranallo Abstract Live attenuated Shigella vaccines have shown promise in inducing protective immune responses in human clinical trials and as carriers of heterologous antigens from other mucosal pathogens. In the past, construction of Shigella vaccine strains relied on classical allelic exchange systems to genetically engineer the bacterial genome. These systems require extensive in vitro engineering of long homologous sequences to create recombinant replication-defective plasmids or phage. Alternatively, the ,red recombination system from bacteriophage facilitates recombination with as little as 40 bp of homologous DNA. The process, referred to as recombineering, typically uses an inducible ,red operon on a temperature-sensitive plasmid and optimal transformation conditions to integrate linear antibiotic resistance cassettes flanked by homologous sequences into a bacterial genome. Recent advances in recombineering have enabled modification of genomic DNA from bacterial pathogens including Salmonella, Yersinia, enteropathogenic Escherichia coli, or enterohemorrhagic E. coli and Shigella. These advances in recombineering have been used to systematically delete virulence-associated genes from Shigella, creating a number of isogenic strains from multiple Shigella serotypes. These strains have been characterized for attenuation using both in vivo and in vitro assays. Based on this data, prototypic Shigella vaccine strains containing multiple deletions in virulence-associated genes have been generated. [source] Inhibition of Listeria monocytogenes in chicken cold cuts by addition of sakacin P and sakacin P-producing Lactobacillus sakeiJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2002T. Katla Aims: To evaluate the potential of sakacin P and sakacin P-producing Lactobacillus sakei for the inhibition of growth of Listeria monocytogenes in chicken cold cuts, by answering the following questions. (i) Is sakacin P actually produced in food? (ii) Is sakacin P produced in situ responsible for the inhibiting effect? (iii) How stable is sakacin P in food? Methods and Results:Listeria monocytogenes, a Lact. sakei strain and/or the bacteriocin sakacin P were added to chicken cold cuts, vacuum packed and incubated at 4 or 10°C for 4 weeks. Each of two isogenic Lact. sakei strains, one producing sakacin P and the other not, had an inhibiting effect on the growth of L. monocytogenes. The effect of these two isogenic strains on the growth of L. monocytogenes was indistinguishable, even though sakacin P was produced in the product by one of the two Lact. sakei strains. The addition of purified sakacin P had an inhibiting effect on the growth of L. monocytogenes. A high dosage of sakacin P (3·5 ,g g,1) had a bacteriostatic effect throughout the storage period of 4 weeks, while a low dosage (12 ng g,1) permitted initial growth, but at a slow rate. After 4 weeks of storage, the number of L. monocytogenes in the samples with a low dosage of sakacin P was 2 logs below that in the untreated control. When using a high dosage of sakacin P, the bacteriocin was detected in samples stored for up to 6 weeks. Conclusions: (i) Sakacin P is produced by a Lact. sakei strain when growing on vacuum-packed chicken cold cuts. (ii) Inhibiting effects of Lact. sakei, other than sakacin P, are active in inhibiting the growth of L. monocytogenes growing on chicken cold cuts. (iii) Sakacin P is stable on chicken cold cuts over a period of 4 weeks. Significance and Impact of the Study: Both sakacin P and Lact. sakei were found to have potential for use in the control of L. monocytogenes in chicken cold cuts. [source] The demography of slow aging in male and female Drosophila mutant for the insulin-receptor substrate homologue chicoAGING CELL, Issue 1 2002Meng-Ping Tu Summary Hypomorphic mutants affecting the Drosophila insulin/IGF signal pathway are reported to increase longevity in females but not in males. To understand this sex-difference, we conducted a large-scale demographic study with three new isogenic strains of alleles at chico, the insulin-receptor substrate homologue. We verify that female dwarf homozygotes (ch1/ch1) and normal-sized heterozygotes (ch1/+) are long-lived, as originally reported. We find for the first time that male heterozygotes are long-lived relative to wildtype, by about 50%. The life span of male ch1/ch1 is similar to that of wildtype but these dwarf males age at a slow demographic rate. The levels of demographic frailty and of age-independent mortality are elevated in ch1/ch1 males, counteracting the effect of slow aging upon life expectancy. Mortality deceleration occurs amongst the oldest-old wildtype adults, as seen in many organisms. Remarkably, in similarly sized cohorts of male and female ch1/ch1 and of male ch1/+ mortality deceleration is absent. Mortality deceleration is a phenotype of chico. [source] Plasma cortisol and metabolite level profiles in two isogenic strains of common carp during confinementJOURNAL OF FISH BIOLOGY, Issue 1 2001N. M. Ruane A rapid increase in common carp Cyprinus carpio plasma cortisol levels was noted, in two experiments, after 30 mins of a 3 h net confinement, which was sustained while the fish were held in the nets. After release from the nets, cortisol levels returned to control values in 1 h. Plasma glucose and free fatty acid levels were elevated by the confinement. Glucose was increased after 30 min but returned to basal levels after 22 h of recovery while free fatty acids were not elevated until 3 h of confinement and remained high for the duration of the recovery period. After confinement for 3 h, plasma lactate levels were reduced and remained low for a further 1 h. No change in either plasma triglyceride or cholesterol levels were found during the study. Confinement had no effect on haematocrit levels but blood haemoglobin levels were reduced. In both experiments hypochloraemia occurred in response to confinement. However, values returned to pre-confinement levels 22 h after confinement. These results show that rearing isogenic carp strains, under identical conditions, results in a reproducable response to an acute stressor and that these carp respond in a similar manner to other teleost species. [source] Efficacy of various pyrethroid structures against a highly metabolically resistant isogenic strain of Helicoverpa armigera (Lepidoptera: Noctuidae) from ChinaPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 10 2007Jianguo Tan Abstract BACKGROUND: Resistance to pyrethroids and other types of insecticides in Helicoverpa armigera (Hübner) has been documented in many countries. The isolation of specific resistance mechanisms in isogenic strains is an optimal approach to investigate cross-resistance pattern, and to validate resistance breaking pyrethroids. In this study an isogenic metabolic resistance CMR strain was successfully isolated from a field pyrethroid-resistant population of H. armigera. With this strain, cross-resistance among 19 pyrethroid insecticides with varying chemical structures was analysed. RESULTS: Resistance to pyrethroids in the CMR strain was likely to be due to enhanced oxidative metabolism. The most significant cross-resistance in the CMR strain was between pyrethroids such as fenvalerate, tau-fluvalinate and flumethrin characterised by having both phenoxybenzyl and aromatic acid moieties. Substitution of the phenoxybenzyl group with a polyfluorobenzyl group, as in tefluthrin, benfluthrin and transfluthrin, overcame most of this resistance. CONCLUSION: The findings in this study support the assertion that it is possible to find pyrethroids that are active against resistant populations. Such pyrethroids could be considered as possible partners or resistance breaking pyrethroids in a pyrethroid resistance management programme for H. armigera in China and in other Asian countries where the oxidative metabolism resistance is a dominant mechanism. Copyright © 2007 Society of Chemical Industry [source] | |||||||||||||