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Selected AbstractsMultiple copies of cytochrome oxidase 1 in species of the fungal genus FusariumMOLECULAR ECOLOGY RESOURCES, Issue 2009SCOTT R. GILMORE Abstract Using data from published mitochondrial or complete genomes, we developed and tested primers for amplification and sequencing of the barcode region of cytochrome oxidase 1 (COX1) of the fungal genus Fusarium, related genera of the order Hypocreales, and degenerate primers for fungi in the subdivision Pezizomycotina. The primers were successful for amplifying and sequencing COX1 barcodes from 13 genera of Hypocreales (Acremonium, Beauveria, Clonostachys, Emericellopsis, Fusarium, Gliocladium, Hypocrea, Lanatonectria, Lecanicillium, Metarhizium, Monocillium, Neonectria and Stilbella), 22 taxa of Fusarium, and two genera in other orders (Arthrosporium, Monilochaetes). Parologous copies of COX1 occurred in several strains of Fusarium. In some, copies of the same length were detected either by heterozygous bases in otherwise clean sequences or in different replicates of amplification and sequencing events; this may indicate multiple transcribed copies. Other strains included one or two introns. Two intron insertion sites had at least two nonhomologous intron sequences among Fusarium species. Irrespective of whether the multiple copy issue could be resolved by sequencing RNA transcripts, developing a precise COX1 -based barcoding system for Fusarium may not be feasible. The overall divergence among homologous COX1 sequences obtained so far is rather low, with many species sharing identical sequences. [source] Cross-protection elicited in channel catfish (Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri strainsAQUACULTURE RESEARCH, Issue 8 2009Victor S Panangala Abstract Cross-protection of channel catfish (Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri and challenged with six E. ictaluri strains was examined in four trials. The relative per cent survival among low-dose immunized and then challenged fish ranged from 27.7% to 100%. Significant protection (P<0.05), with the exception of strain ATCC-33202, was conferred by immunization with a given E. ictaluri strain challenged either with a homologous or a heterologous strain. Antibody titres of pooled serum collected on day 22 from surviving fish examined by enzyme-linked immunosorbent assay (ELISA) ranged from 1:40 to 1:320, but no differences were apparent among different vaccinated groups. The protein profiles of six E. ictaluri strains examined by Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a relatively homogeneous pattern. Immuno-blots probed with pooled serum from immunized and challenged fish showed a pattern similar to LPS-reaction patterns observed with E. ictaluri in other studies. Since the present studies further corroborate that E. ictaluri is a clonal bacterial species with no apparent antigenic differences, it is possible that immunization with a single E. ictaluri field strain should confer protection against any other strain. [source] Isolation and properties of methanesulfonate-degrading Afipia felis from Antarctica and comparison with other strains of A. felisENVIRONMENTAL MICROBIOLOGY, Issue 1 2005S. Azra Moosvi Summary Three novel strains of methylotrophic Afipia felis were isolated from several locations on Signy Island, Antarctica, and a fourth from estuary sediment from the River Douro, Portugal. They were identified as strains of the ,-2 proteobacterium A. felis by 16S rRNA gene sequence, analysis., Two, strains, tested, were, shown to contain the fdxA gene, diagnostic for A. felis. All strains grew with methanesulfonate (and two strains with dimethylsulfone) as sole carbon substrate. Growth on methanesulfonate required methanesulfonate monooxygenase (MSAMO), using NADH as the reductant and stimulated by reduced flavin nucleotides and Fe(II). Polymerase chain reaction amplification of DNA from an Antarctic strain showed a typical msmA gene for the ,-hydroxylase of MSAMO, and both Antarctic and Portuguese strains contained mxaF, the methanol dehydrogenase large subunit gene. This is the first report of methanesulfonate-degrading bacteria from the Antarctic and of methylotrophy in Afipia, and the first description of any bacterium able to use both methanesulfonate and dimethylsulfone. In contrast, the type strain of A. felis DSM 7326T was not methylotrophic, but grew in defined mineral medium with a wide range of single simple organic substrates. Free-living Afipia strains occurring widely in the natural environment may be significant as methylotrophs, degrading C1 -sulfur compounds, including the recalcitrant organosulfur compound methanesulfonate. [source] Microevolutionary analysis of the nematode genus Pristionchus suggests a recent evolution of redundant developmental mechanisms during vulva formationEVOLUTION AND DEVELOPMENT, Issue 4 2001Jagan Srinivasan SUMMARY To identify the mechanisms by which molecular variation is introduced into developmental systems, microevolutionary approaches to evolutionary developmental biology have to be taken. Here, we describe the molecular and developmental characterization of laboratory strains of the nematode genus Pristionchus, which lays a foundation for a microevolutionary analysis of vulva development. We describe 13 laboratory strains of the Pristionchus genus that are derived from natural isolates from around the world. Mating experiments and ITS sequence analysis indicated that these 13 strains represent four different species: the gonochoristic species P. lheritieri and three hermaphroditic species, P. pacificus, P. maupasi, and an as yet undescribed species Pristionchus sp., respectively. P. pacificus is represented by five different strains isolated from California, Washington, Hawaii, Ontario, and Poland. Developmental differences during vulva formation are observed between strains from different species but also between strains of P. pacificus, like the strains from California and Poland. In particular, redundant developmental mechanisms present during vulva formation in P. pacificus var. California are absent in other strains. Amplified restriction fragment length polymorphism (AFLP) analyses of the P. pacificus strains revealed that the American strains are highly polymorphic. In contrast, the developmentally distinct strain from Poland is identical to the Californian strain, suggesting that the developmental differences rely on a small number of changes in developmental control genes rather than the accumulation of changes at multiple loci. [source] Growth of Frankia strains in leaf litter-amended soil and the rhizosphere of a nonactinorhizal plantFEMS MICROBIOLOGY ECOLOGY, Issue 1 2009Babur S. Mirza Abstract The ability of Frankia strains to grow in the rhizosphere of a nonactinorhizal plant, Betula pendula, in surrounding bulk soil and in soil amended with leaf litter was analyzed 6 weeks after inoculation of pure cultures by in situ hybridization. Growth responses were related to taxonomic position as determined by comparative sequence analysis of nifH gene fragments and of an actinomycetes-specific insertion in Domain III of the 23S rRNA gene. Phylogenetic analyses confirmed the basic classification of Frankia strains by host infection groups, and allowed a further differentiation of Frankia clusters within the Alnus host infection group. Except for Casuarina -infective Frankia strains, all other strains of the Alnus and the Elaeagnus host infection groups displayed growth in the rhizosphere of B. pendula, and none of them grew in the surrounding bulk soil that was characterized by a very low organic matter content. Only a small number of strains that all belonged to a distinct phylogenetic cluster within the Alnus host infection group grew in soil amended with ground leaf litter from B. pendula. These results demonstrate that saprotrophic growth of frankiae is a common trait for most members of the genus, and the supporting factors for growth (i.e. carbon utilization capabilities) varied with the host infection group and the phylogenetic affiliation of the strains. [source] Evidence for a female-specific effect of a chromosome 4 locus on anxiety-related behaviors and ethanol drinking in ratsGENES, BRAIN AND BEHAVIOR, Issue 6 2006L. F. Vendruscolo Previous studies using the inbred rat strains Lewis (LEW) and spontaneously hypertensive rats (SHR) led to the mapping of two quantitative trait loci, named Ofil1 (on chromosome 4 of the rat) and Ofil2 (on chromosome 7), for open-field inner locomotion, a behavioral index of anxiety. Studies using other strains showed that the region next to Ofil1 influences measures of not only anxiety but also ethanol consumption. In view of the high prevalence of psychiatric disorders such as anxiety and alcoholism, as well as the comorbidity between them, the present study was designed to better characterize the contribution of these two loci to complex emotional and consummatory responses. Rats deriving from an F2 intercross between the LEW and the SHR strains were selected according to their genotype at markers flanking the loci Ofil1 and Ofil2 and bred to obtain lines of rats homozygous LEW/LEW or SHR/SHR for each of the two loci, thus generating four genotypic combinations. These selected animals as well as purebred LEW and SHR rats of both sexes were submitted to a battery of tests including measures of locomotor activity, anxiety, sweet and bitter taste reinforcement and ethanol intake. Lewis rats displayed more anxiety-like behavior and less ethanol intake than SHR rats. Ofil1 (on chromosome 4) affected both the activity in the center of the open field and ethanol drinking in females only. These results suggest that Ofil1 contains either linked genes with independent influences on anxiety-related responses and ethanol drinking or a pleiotropic gene with simultaneous effects on both traits. [source] Impact of heat shock on heat shock proteins expression, biological and commercial traits of Bombyx moriINSECT SCIENCE, Issue 4 2006VASUDHA B. CHAVADI Abstract We report the thermotolerance of new bivoltine silkworm, Bombyx mori strains NB4D2, KSO1, NP2, CSR2 and CSR4and differential expression of heat shock proteins at different instars. Different instars of silkworm larva were subjected to heat shock at 35°C, 40°C and 45°C for 2 hours followed by 2 hours recovery. Heat shock proteins were analyzed by SDS-PAGE. The impact of heat shock on commercial traits of cocoons was analyzed by following different strategies in terms of acquired thermotolerance over control. Comparatively NP2 exhibited better survivability than other strains. Resistance to heat shock was increased as larval development proceeds in the order of first instar > second instar > third instar > fourth instar > fifth instar in all silkworm strains. Expression of heat shock proteins varies in different instars. 90 kDa in the first, second and third instars, 84 kDa in the fourth instar and 84, 62, 60, 47 and 33 kDa heat shock proteins in fifth instar was observed in response to heat shock. Relative influence of heat shock on commercial traits that correspond to different stages was significant in all strains. In NB4D2, cocoon and shell weight significantly increased to 17.52% and 19.44% over control respectively. Heat shock proteins as molecular markers for evaluation and evolution of thermotolerant silkworm strains for tropics was discussed. [source] Computational studies of electron-transfer processes in old yellow enzymeINTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY, Issue 6 2001Ginger M. Chateauneuf Abstract Old Yellow Enzyme (OYE) is a flavoenzyme that was first isolated from brewer's bottom yeast. Homologues have been identified in other strains of yeast, bacteria, and plants. In plants, the OYE homologue functions enzymatically in the synthesis of plant hormones, but the biological function of OYE in yeast is still unknown. Flavin mononucleotide (FMN) is the cofactor that is noncovalently bound in the enzyme. OYE binds several phenolic ligands that serve as models for reactive biological substrates. These complexes have broad long-wavelength absorption bands, which have been ascribed to charge-transfer interactions, with the phenolate anion acting as the electron donor and the FMN as the acceptor [Abramovitz, A. S.; Massey, V. J Bio Chem 1976, 251, 5327,5336]. The computational characterization of these electronic transitions in the active site will help in understanding the biological processes in the enzyme. It was found that at several levels of computational methods, and through computationally mutating relevant amino acids, a charge-transfer process is occurring. This result agrees with previous experimental work and is consistent with all ultraviolet,visible spectrophotometric data. The preliminary results for the computational studies of these electron-transfer processes will be presented. © 2001 John Wiley & Sons, Inc. Int J Quantum Chem, 2001 [source] Optimization of agro-residual medium for ,-amylase production from a hyper-producing Bacillus subtilis KCC103 in submerged fermentationJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 4 2009Gobinath Rajagopalan Abstract BACKGROUND: Although submerged fermentation (SmF) is the conventional method in industry, use of low-cost agro-residues for ,-amylase production in SmF has not been well established. Here we optimized agro-residue-based medium and culture conditions for ,-amylase production in SmF using a hyper-producing Bacillus subtilis KCC103. RESULTS:B. subtilis KCC103 produced ,-amylase in SmF by utilizing agro-residues. Wheat bran (WB) and sunflower oil cake (SFOC) were selected as the best substrates using shake flasks. Medium containing WB (carbohydrate rich) and SFOC (rich in protein and free amino acids) at 1:1 (w/w) ratio produced high levels (90 IU mL,1) of ,-amylase at 30,36 h in a shake flask. The ,-amylase yield was 14-fold enhanced (1258 IU mL,1) by optimizing process parameters and medium composition following response surface methodology in a bioreactor. The optimal conditions were: WB 1.27%, SFOC 1.42%, pH 7, 37 °C and 10,12 h. Both in shake flask and bioreactor ,-amylase synthesis was not repressed by the release of simple sugars into the medium. CONCLUSION: KCC103 with catabolite derepression and hyperproducing ability is useful for economic ,-amylase production using low-cost agro-residual substrates in conventional SmF. Since the production time (10,12 h) is much shorter than other strains this would improve productivity and further reduce the cost of ,-amylase production. Copyright © 2008 Society of Chemical Industry [source] Differential cytokine expression by human dendritic cells in response to different Porphyromonas gingivalis capsular serotypesJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 10 2009Rolando Vernal Abstract Aim: Capsular polysaccharides play an important role in the virulence of Gram-positive and Gram-negative bacteria. In Porphyromonas gingivalis, six serotypes have been described based on capsular antigenicity and its pathogenicity has been correlated both in vitro and in animal models. This study aimed to investigate the differential response of human dendritic cells (DCs) when stimulated with different P. gingivalis capsular serotypes. Materials and Methods: Using different multiplicity of infection (MOI) of the encapsulated strains K1,K6 and the non-encapsulated K, strain of P. gingivalis, the mRNA expression levels for interleukin (IL)-1,, IL-2, IL-5, IL-6, IL-10, IL-12, IL-13, interferon (IFN)- ,, tumour necrosis factor (TNF)- ,, and TNF- , in stimulated DCs were quantified by real-time reverse transcription-polymerase chain reaction. Results: All P. gingivalis capsular serotypes induced a T-helper type 1 (Th1) pattern of cytokine expression. K1- and K2-stimulated DCs expressed higher levels of IL-1,, IL-6, IL-12p35, IL-12p40, and IFN- , and at lower MOI than DCs stimulated with the other strains. Conclusions: These results demonstrate a differential potential of P. gingivalis capsular serotypes to induce DC responses and a higher capacity of strains K1 W83 and K2 HG184 than other K serotypes to trigger cytokine expression. [source] Violent phenotype in SAL mice is inflexible and fixed in adulthoodAGGRESSIVE BEHAVIOR, Issue 5 2009Deepa Natarajan Abstract Violence was shown to be qualitatively different from functional hyper-aggression in mice selected for high aggression namely Short Attack Latency (SAL), Turku Aggressive (TA) and North Carolina (NC900) strains. This study aimed at investigating whether this adulthood violent phenotype as seen previously in the SAL mice is fixed and hence behaviorally inflexible right from day 1 of the experiment or consequential, i.e., subject to gradual change from functional aggression to violence. The functionally hyper-aggressive strains namely TA and NC900 strains served as controls for the study. Methodologically, behavioral (in)flexibility was studied using the overall sequential structure of agonistic behavior. In particular, intra-individual variations in the overall agonistic behavior as well as offensive, pre- and post-offensive behavior transitions, directly related to the resident,intruder interactions were investigated. The SAL mice showed the least intra-individual variation in their overall sequential agonistic structure as well as a fixed offense-oriented agonistic behavior of highest magnitude when compared with the other strains. Additionally, the pre- and post- offensive transitions were most salient in the functionally hyper-aggressive TA and NC900 strains, whereas virtually absent in the SAL mice. Thus, the violent behavior of the adult SAL mice is behaviorally inflexible or fixed, whereas the functionally hyper-aggressive behavior of the adult TA and NC900 mice is behaviorally flexible and constantly adaptive to the opponent behavior, over 3 days of repeated resident,intruder interaction. Aggr. Behav. 35:430,436, 2009. © 2009 Wiley-Liss, Inc. [source] Structural characteristics of the cag pathogenicity island and its significance in the classification of Chinese strains of Helicobacter pyloriJOURNAL OF DIGESTIVE DISEASES, Issue 2 2002Jiong LIU OBJECTIVE: To investigate the structural characteristics of the cag pathogenicity island (PAI) and its significance in the classification in Chinese strains of Helicobacter pylori. METHODS: In 107 H. pylori strains isolated from Chinese patients, cagA, cagI, cagII, the cagI,cagII junction and IS605 were studied by using the polymerase chain reaction. RESULTS: The positive rates in Chinese H. pylori strains were 95.3% for cag PAI, 92.5% for cagA, 86.9% for cagI and 66.4% for cagII. There was no statistical difference among H. pylori strains from chronic gastritis, peptic ulcers or gastric carcinoma in the detectable rate of cag PAI, cagA, cagI or cagII (P > 0.05). Of the cag PAI-negative strains, four came from cases of chronic gastritis and one from a patient with cardiac cancer. The products of the cagI,cagII junction were found in only five strains. The continuous cag PAI was much more common in duodenal ulcers than in chronic gastritis (P < 0.01). The positive rates of cagI and cagII were markedly different in chronic gastritis (P < 0.05). One strain of H. pylori tested positive for cagA but negative for other regions of the cag PAI. IS605 was less common in duodenal ulcers than in chronic gastritis (P < 0.05). The amplified fragment of IS605 in one strain from a gastric carcinoma was approximately 1580 bp in size, which was much longer than that in other strains. CONCLUSION: Our results indicate that the cag PAI is very common in Chinese strains of H. pylori. The structural variety of the cag PAI might be related to the virulence of H. pylori. It is suggested that H. pylori may be classified into different virulence groups according to differences in the structure of the cag PAI. [source] Phylogenetic analyses and molecular epidemiology of European salmonid alphaviruses (SAV) based on partial E2 and nsP3 gene nucleotide sequencesJOURNAL OF FISH DISEASES, Issue 11 2008E Fringuelli Abstract Sequence data were generated for portions of the E2 and nsP3 genes of 48 salmonid alphaviruses from farmed Atlantic salmon (AS), Salmo salar L., and rainbow trout (RT), Oncorhynchus mykiss (Walbaum), in marine and freshwater environments, respectively, from the Republic of Ireland, Northern Ireland, England, Scotland, Norway, France, Italy and Spain between 1991 and 2007. Based on these sequences, and those of six previously published reference strains, phylogenetic trees were constructed using the parsimony method. Trees generated with both gene segments were similar. Clades corresponding to the three previously recognized subtypes were generated and in addition, two further new clades of viruses were identified. A single further strain (F96-1045) was found to be distinct from all of the other strains in the study. The percentage of nucleotide divergence within clades was generally low (0,4.8% for E2, 0,6.6% for nsP3). Interclade divergence tended to be higher (3.4,19.7% for E2, 6.5,28.1% for nsP3). Based on these results and using current SAV terminology, the two new clades and F96-1045 were termed SAV subtypes 4, 5 and 6, respectively. SAV4 contained AS strains from Ireland and Scotland, while SAV5 contained only Scottish AS strains. Recently identified SAV strains from RT in Italy and Spain were shown to belong to SAV2. In addition, marine AS strains belonging to SAV2 were identified for the first time. Analysis of the origin of several clusters of strains with identical E2 and nsP3 sequences strongly support horizontal transmission of virus between farms and aquaculture companies. Evidence in support of vertical transmission was not found. [source] VARIATION OF LAG TIME AND SPECIFIC GROWTH RATE AMONG 11 STRAINS OF SALMONELLA INOCULATED ONTO STERILE GROUND CHICKEN BREAST BURGERS AND INCUBATED AT 25C,JOURNAL OF FOOD SAFETY, Issue 4 2000THOMAS P. OSCAR ABSTRACT One strain of 11 serotypes or 11 strains of Salmonella, which were isolated from the ceca of broilers, were surveyed for their growth kinetics on sterile ground chicken breast burgers incubated at 25C to determine the variation of lag time and specific growth rate. Growth curves, four per strain, were fit to a two-phase linear model to determine lag time (h) and specific growth rate (log10/h). Repeatability of growth kinetics measurements for individual strains had a mean coefficient of variation of 11.7% for lag time (range: 5.8 to 17.3%) and a mean coefficient of variation of 6.7% for specific growth rate (range: 2.7 to 13.3%). Lag time among strains ranged from 2.2 to 3.1 h with a mean of 2.8 h for all strains, whereas specific growth rate among strains ranged from 0.3 to 0.38 log10 per h with a mean of 0.35 log10per h for all strains. One-way analysis of variance indicated that lag time (P =0.029) and specific growth rate (P =0.025) differed slightly among strains. S. Haardt had a shorter (P < 0.05) lag time than S. Agona and S. Brandenburg, whereas the specific growth rate of S. Enteritidis was less than (P < 0.05) the specific growth rates of S. Typhimurium and S. Brandenburg. All other strains had similar lag times and specific growth rates. The coefficient of variation among strains was 9.4% for lag time and 5.7% for specific growth rate. These results indicate that there were only minor differences in the lag times and specific growth rates among the strains of Salmonella surveyed. Thus, the growth kinetic values obtained with one strain of Salmonella may be useful for predicting the growth of other strains of Salmonella for which data do not currently exist. [source] Heritability of the Blood Pressure Response to Acute Ethanol Exposure in Five Inbred Strains of MiceALCOHOLISM, Issue 10 2000Daniel C. Hatton Background: Chronic alcohol consumption is a major risk factor for hypertension. There is evidence in humans that the susceptibility to alcohol-related hypertension may vary based on genotype. As a first step in investigating the genetic basis for alcohol-related hypertension, the current study was designed to assess the heritability of the blood pressure response to acute ethanol exposure by using AKR/J (AK), C57BL/6J (B6), DBA/2J (D2), Balb/cJ (Balb), and A/J (A) mice. Methods: Mean arterial pressure (MAP) was recorded continuously for 24 hr in freely moving mice from an indwelling femoral catheter before we tested the effects of saline or ethanol (2 g/kg ip) on blood pressure. Results: Relative to saline, ethanol caused a pressor response that peaked within 10 min, followed by a decline in MAP. Strain A mice had a significantly greater pressor response to ethanol than other strains and did not show a decline in MAP below baseline. All other strains showed a progressive fall in blood pressure below baseline across the 60 min measurement interval. Heritability was estimated to be 0.62 for the pressor response and 0.64 for the maximal depressor response. Repeated doses of ethanol at 1 hr intervals in A and B6 mice (0,2,1.5,1.5,1.5 g/kg ip) resulted in a dose-dependent increase in MAP in A mice for the first three doses and a dose-dependent decrease in MAP in B6 mice that was independent of blood ethanol concentrations. Conclusion: The results indicate that there is a significant genetic component to the acute blood pressure response to ethanol. [source] Binding of extracellular matrix molecules by probiotic bacteriaLETTERS IN APPLIED MICROBIOLOGY, Issue 4 2003tyriak Abstract Aims: The aim of this study was to investigate extracellular matrix (ECM) and mucin binding of selected bacterial isolates with probiotic features in comparison with commercially used probiotic bacteria. Methods and Results: ECM molecules were immobilized in microtitre plates (mucin and fetuin) or on the surface of latex beads. Porcine mucin was bound by all 13 probiotic strains tested with important inter-strain differences; however, fetuin binding was similar (weak) for all 14 strains tested. Strongly positive (three) binding of bovine fibrinogen was expressed by strains from fermented food (Lactobacillus rhamnosus GG, L. casei Shirota and L. johnsonii La1) as well as by L. casei L.c., Lactobacillus sp. 2I3 and by L. plantarum LP. The other strains expressed moderate (2) or weakly positive (1) binding of bovine fibrinogen. Strongly positive (3) binding of porcine fibronectin was observed only with two strains; however, all other strains also bound this molecule. Bovine lactoferrin was bound to a higher extent than transferrins. Significance and Impact of the Study: Some animal strains (at least L. casei L.c. and Lactobacillus sp. 2I3) are comparable with the commercially used strains with respect to their ECM binding ability. As this feature is important for probiotic bacteria to be able to colonize intestine, these strains should be considered for their wider use in fermented feed (or probiotic preparations) for animals. [source] Pyrethroid and DDT cross-resistance in Aedes aegypti is correlated with novel mutations in the voltage-gated sodium channel geneMEDICAL AND VETERINARY ENTOMOLOGY, Issue 1 2003C. Brengues Abstract. Samples of the dengue vector mosquito Aedes aegypti (L.) (Diptera: Culicidae) were collected from 13 localities between 1995 and 1998. Two laboratory strains, Bora (French Polynesia) and AEAE, were both susceptible to DDT and permethrin; all other strains, except Larentuka (Indonesia) and Bouaké (Ivory Coast), contained individual fourth-instar larvae resistant to permethrin. Ten strains were subjected to a range of biochemical assays. Many strains had elevated carboxylesterase activity compared to the Bora strain; this was particularly high in the Indonesian strains Salatiga and Semarang, and in the Guyane strain (Cayenne). Monooxygenase levels were increased in the Salatiga and Paea (Polynesia) strains, and reduced in the two Thai strains (Mae Kaza, Mae Kud) and the Larentuka strain. Glutathione S-transferase activity was elevated in the Guyane strain. All other enzyme profiles were similar to the susceptible strain. The presence of both DDT and pyrethroid resistance in the Semarang, Belem (Brazil) and Long Hoa (Vietnam) strains suggested the presence of a knock-down resistant (kdr)-type resistance mechanism. Part of the S6 hydrophobic segment of domain II of the voltage-gated sodium channel gene was obtained by RT-PCR and sequenced from several insects from all 13 field strains. Four novel mutations were identified. Three strains contained identical amino acid substitutions at two positions, two strains shared a different substitution, and one strain was homozygous for a fourth alteration. The leucine to phenylalanine substitution that confers nerve insensitivity to pyrethroids in a range of other resistant insects was absent. Direct neurophysiological assays on individual larvae from three strains with these mutations demonstrated reduced nerve sensitivity to permethrin or lambda cyhalothrin inhibition compared to the susceptible strains. [source] Strain-specific regulation of intracellular Wolbachia density in multiply infected insectsMOLECULAR ECOLOGY, Issue 12 2003L. Mouton Abstract Vertically transmitted symbionts suffer a severe reduction in numbers when they pass through host generations, resulting in genetic homogeneity or even clonality of their populations. Wolbachia endosymbionts that induce cytoplasmic incompatibility in their hosts depart from this rule, because cytoplasmic incompatibility actively maintains multiple infection within hosts. Hosts and symbionts are thus probably under peculiar selective pressures that must shape the way intracellular bacterial populations are regulated. We studied the density and location of Wolbachia within adult Leptopilina heterotoma, a haplodiploid wasp that is parasitic on Drosophila and that is naturally infected with three Wolbachia strains, but for which we also obtained one simply infected and two doubly infected lines. Comparison of these four lines by quantitative polymerase chain reaction using a real-time detection system showed that total Wolbachia density varies according to the infection status of individuals, while the specific density of each Wolbachia strain remains constant regardless of the presence of other strains. This suggests that Wolbachia strains do not compete with one another within the same host individual, and that a strain-specific regulatory mechanism is operating. We discuss the regulatory mechanisms that are involved, and how this process might have evolved as a response to selective pressures acting on both partners. [source] Evolution of mutation rates in bacteriaMOLECULAR MICROBIOLOGY, Issue 4 2006Erick Denamur Summary Evolutionary success of bacteria relies on the constant fine-tuning of their mutation rates, which optimizes their adaptability to constantly changing environmental conditions. When adaptation is limited by the mutation supply rate, under some conditions, natural selection favours increased mutation rates by acting on allelic variation of the genetic systems that control fidelity of DNA replication and repair. Mutator alleles are carried to high frequency through hitchhiking with the adaptive mutations they generate. However, when fitness gain no longer counterbalances the fitness loss due to continuous generation of deleterious mutations, natural selection favours reduction of mutation rates. Selection and counter-selection of high mutation rates depends on many factors: the number of mutations required for adaptation, the strength of mutator alleles, bacterial population size, competition with other strains, migration, and spatial and temporal environmental heterogeneity. Such modulations of mutation rates may also play a role in the evolution of antibiotic resistance. [source] Modulation of pPS10 host range by DnaAMOLECULAR MICROBIOLOGY, Issue 1 2002Beatriz Maestro Summary Narrow-host-range plasmid pPS10, originally found in Pseudomonas savastanoi, is unable to replicate in other strains such as Escherichia coli. Here, we report that the establishment of pPS10 in E. coli can be achieved by a triple mutation in the dnaA gene of E. coli (dnaA403), leading to Q14amber, P297S and A412V changes in the DnaA host replication protein (DnaA403 mutant). As the E. coli strain used contained double amber suppressor mutations (supE, supF), the amber codon in dnaA403 can be translated into glutamine or tyrosine. Genetic analysis of DnaA proteins containing either the individual changes or their different combinations suggests that the P297S mutation is crucial for the establishment of the pPS10 replicon in E. coli. The data also indicate that the P297S change is toxic to the cell and that the additional mutations in DnaA403 could contribute to neutralize this toxicity. To our knowledge, this work reports the first chromosome mutant described in the literature that allows the host range broadening of a plasmid, highlights the essential role played by DnaA in the establishment of pPS10 replicon in E. coli and provides support for the hypothesis that interactions between RepA and DnaA modulate the establish-ment of pPS10 in that bacteria and probably in other species. [source] Excision from tRNA genes of a large chromosomal region, carrying avrPphB, associated with race change in the bean pathogen, Pseudomonas syringae pv. phaseolicolaMOLECULAR MICROBIOLOGY, Issue 2 2000Robert W. Jackson Pseudomonas syringae pv. phaseolicola (Pph) race 4 strain 1302A carries avirulence gene avrPphB. Strain RJ3, a sectoral variant from a 1302A culture, exhibited an extended host range in cultivars of bean and soybean resulting from the absence of avrPphB from the RJ3 chromosome. Complementation of RJ3 with avrPphB restored the race 4 phenotype. Both strains showed similar in planta growth in susceptible bean cultivars. Analysis of RJ3 indicated loss of >,40 kb of DNA surrounding avrPphB. Collinearity of the two genomes was determined for the left and right junctions of the deleted avrPphB region; the left junction is ,,19 kb and the right junction >,20 kb from avrPphB in 1302A. Sequencing revealed that the region containing avrPphB was inserted into a tRNALYS gene, which was re-formed at the right junction in strain 1302A. A putative lysine tRNA pseudogene (,tRNALYS) was found at the left junction of the insertion. All tRNA genes were in identical orientation in the chromosome. Genes near the left junction exhibited predicted protein homologies with gene products associated with a virulence locus of the periodontal pathogen Actinobacillus actinomycetemcomitans. Specific oligonucleotide primers that differentiate 1302A from RJ3 were designed and used to demonstrate that avrPphB was located in different regions of the chromosome in other strains of Pph. Deletion of a large region of the chromosome containing an avirulence gene represents a new route to race change in Pph. [source] In vitro fermentability of human milk oligosaccharides by several strains of bifidobacteriaMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 11 2007Robert E. Ward Abstract This study was conducted to investigate the catabolism and fermentation of human milk oligosaccharides (HMO) by individual strains of bifidobacteria. Oligosaccharides were isolated from a pooled sample of human milk using solid-phase extraction, and then added to a growth medium as the sole source of fermentable carbohydrate. Of five strains of bifidobacteria tested (Bifidobacterium longum biovar infantis, Bifidobacterium bifidum, Bifidobacterium longum biovar longum, Bifidobacterium breve, and Bifidobacterium adolescentis), B. longum bv. infantis grew better, achieving triple the cell density then the other strains. B. bifidum did not reach a high cell density, yet generated free sialic acid, fucose and N-acetylglucosamine in the media, suggesting some capacity for HMO degradation. Thin layer chromatography profiles of spent fermentation broth suggests substantial degradation of oligosaccharides by B. longum bv. infantis, moderate degradation by B. bifidum and little degradation by other strains. While all strains were able to individually ferment two monosaccharide constituents of HMO, glucose and galactose, only B. longum bv. infantis and B. breve were able to ferment glucosamine, fucose and sialic acid. These results suggest that as a potential prebiotic, HMO may selectively promote the growth of certain bifidobacteria strains, and their catabolism may result in free monosaccharides in the colonic lumen. [source] Differential effects of five Aggregatibacter actinomycetemcomitans strains on gingival epithelial cellsMOLECULAR ORAL MICROBIOLOGY, Issue 6 2008T. Shimada Introduction:, We investigated gingival epithelial cell proliferation and expression of interleukin-8 (IL-8) and intercellular adhesion molecule 1 (ICAM-1) in response to Aggregatibacter actinomycetemcomitans serotypes a, b, and c. Methods:, Human gingival cells (Ca9-22) were cultured in bacterial extracts prepared from five strains of A. actinomycetemcomitans: ATCC 43717 (serotype a); ATCC 29524, ATCC 29522, and ATCC 43718 (all serotype b); and ATCC 43719 (serotype c). Results:, In bacterial extracts of ATCC 29522, cell growth was significantly impaired, while the expression of IL-8 and ICAM-1 was significantly increased. The level of induction in response to the other strains was minimal. Conclusion:, Our results indicate that the five strains of A. actinomycetemcomitans have distinct effects on the abilities of human gingival epithelial cells to proliferate and to produce proinflammatory factors. [source] Deletion in sortase gene of Streptococcus mutans IngbrittMOLECULAR ORAL MICROBIOLOGY, Issue 3 2004T. Igarashi Our previous studies on Streptococcus mutans have demonstrated that surface proteins containing a C-terminal sorting signal, such as surface protein antigen (PAc), glucan-binding protein C (GbpC) and dextranase (Dex), are anchored to the cell wall by a sortase (SrtA). In this study we found that, unlike other strains of S. mutans, strain Ingbritt did not exhibit cell wall-anchoring of PAc, GbpC and Dex. It is speculated that the SrtA of strain Ingbritt did not function in the cell wall-anchoring process of these surface proteins. Sequence analysis revealed a deletion of an 11-bp nucleotide sequence in the srtA gene of strain Ingbritt, resulting in the generation of a new termination codon, resulting in production of an incomplete SrtA enzyme protein. As a result, strain Ingbritt showed a localization change of PAc, GbpC and Dex in the cell, implying that strain Ingbritt loses the biological functions mediated by the cell surface-associated proteins of S. mutans. These results suggest that strain Ingbritt could be less cariogenic than other strains of S. mutans. [source] Probiotics and health: a review of the evidenceNUTRITION BULLETIN, Issue 4 2009E. Weichselbaum Summary Probiotics are live microorganisms , mainly bacteria , which when administered in adequate amounts confer a health benefit on the host. There is rising interest in this area, but reports in the media are often conflicting. The aim of this review is to consider the current evidence on the effects of probiotics on health, focusing on gut-related health issues and the immune system, with the objective to provide a clearer picture of whether and how probiotics can be beneficial for health. The outcomes of this review are based on more than 100 original studies, meta-analyses and systematic reviews. A variety of different strains have been used in studies on probiotics, and it is important to remember that the effectiveness of probiotics is strain-specific, which means that each single probiotic strain has to be tested to assess its potential health benefits. Overall, despite the diversity of strains used in the studies included in this review, there is evidence that probiotics have the potential to be beneficial for our health. Studies in patients with inflammatory bowel disease show probiotic strains to be able to decrease the recurrence of ulcerative colitis and occurrence and recurrence of pouchitis, however, current evidence suggests that probiotics are ineffective in treating patients with Crohn's disease. Patients with irritable bowel syndrome show a reduction in symptoms when treated with selected probiotic strains, but high placebo effects have been reported as well. The evidence of the efficacy of probiotics in patients suffering from constipation is limited, but the evidence seems promising for some strains to bring relief to patients suffering from constipation. There is good evidence that a number of probiotic strains are effective in preventing antibiotic-associated diarrhoea. The most commonly studied strains are Lactobacillus rhamnosus GG (LGG) and Saccharomyces boulardii, but other strains and mixtures of strains seem to be effective as well. There is also promising evidence of a preventive effect of probiotics in Clostridium difficile -associated diarrhoea, although some studies have been too small to obtain statistically significant findings. The effect of probiotics in acute diarrhoea, particularly in children, is well studied. Selected probiotic strains seem to be effective in reducing the duration of acute diarrhoea. LGG and S. boulardii are again the most commonly used strains and a number of studies have shown them to be effective, although one meta-analysis showed that the effect of LGG was only significant in children in Western countries, not in children in developing countries, which may be due to different causes of diarrhoea in these regions. Studies investigating the preventive effect of probiotics in the context of common cold and flu infections show that the studied strains failed to lower the incidence of episodes but that they have the potential to decrease the duration of episodes, which suggests that the immune system may be more efficient in fighting off common cold and flu infections after consuming these strains. The evidence so far does not suggest that probiotics are effective in preventing or treating allergies or in treating eczema. However, some probiotic strains seem to lower the risk of developing eczema if taken by pregnant women and their infants in early life. [source] New developments in insecticide resistance in the glasshouse whitefly (Trialeurodes vaporariorum) and the two-spotted spider mite (Tetranychus urticae) in the UKPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 2 2002Kevin Gorman Abstract A recent survey of insecticide resistance in two of the most problematic pests in UK glasshouses revealed some new developments. At least some individuals in all UK samples of Trialeurodes vaporariorum that were tested resisted the insect growth regulator (IGR) buprofezin. The most strongly resistant strains were unaffected by the field application rate of this compound, and even samples from populations that had never been exposed to buprofezin contained individuals that survived the highest concentration applied (10,000,mg,litre,1). The field rate of buprofezin was shown to select for resistance through vapour action alone. The benzophenylurea teflubenzuron, an unrelated IGR, was cross-resisted by buprofezin-resistant individuals. There was no evidence of resistance to imidacloprid, but all T vaporariorum strains tested, regardless of origin, exhibited a high innate tolerance to nicotine, when compared with another whitefly species, Bemisia tabaci. Marked resistance to fenbutatin oxide and tebufenpyrad was found in single glasshouse populations of Tetranychus urticae, but these compounds and abamectin appeared to remain highly effective against all other strains collected. © 2001 Society of Chemical Industry [source] Simple assay for antitumour immunoactive glycoprotein derived from Chlorella vulgaris strain CK22 using ELISAPHYTOTHERAPY RESEARCH, Issue 6 2002Kiyoshi Noda Abstract A quantitative ELISA system was developed using a monoclonal antibody (MAb) specific for an antitumour immunoactive glycoprotein (CVS) derived from C. vulgaris strain CK22. The full measuring range of the assay extends from 0.63 to 10.0,ng/mL of CVS. Although no cross-reaction was observed to proteins tested or other biological response modifiers (BRMs) derived from different sources, cross-reactions were found with culture supernatants from two other strains of C. vulgaris having a strong antitumour immunoactivity. Treatment of CVS with protease, acid or alkali weakened or completely eliminated the reactivity against the MAb and also its antitumour immunoactivities. This ELISA system is suitable for the biologically active form of CVS derived from C. vulgaris strain CK22 and related immunoactive strains. Copyright © 2002 John Wiley & Sons, Ltd. [source] Vegetative compatibility and heterokaryon stability in Fusarium oxysporum f.sp. radicis-lycopersici from ItalyPLANT PATHOLOGY, Issue 3 2001P. Di Primo Fusarium crown and root rot, caused by Fusarium oxysporum f.sp. radicis-lycopersici (Forl), is one of the most destructive soilborne diseases of tomato in Italy. Chlorate-resistant, nitrate-nonutilizing (nit) mutants were used to determine vegetative compatibility among 191 isolates of Forl collected in five geographic regions (Calabria, Emilia-Romagna, Liguria, Sardinia, Sicily) in Italy. The isolates were assigned to five vegetative compatibility groups (VCGs): 65 isolates to VCG 0090; 99 to VCG 0091; 23 to VCG 0092; two to VCG 0093; and two to VCG 0096. The population structure of Forl in Italy is similar to that reported for Israel, and differs from that found in North Atlantic European countries, where VCG 0094 is predominant. The stability of prototrophic heterokaryons originating from hyphal anastomosis between compatible complementary nit mutants was assessed through conidial analysis and mycelial mass transfer. Most monoconidial cultures (84%) recovered from 117 prototrophic heterokaryons were nit mutants, indicating that heterokaryons generally do not proliferate well through conidiation; most of the 177 prototrophic heterokaryons examined were unstable, and only 9% sustained prototrophic growth through the tenth mycelial transfer upon subculturing. The prototrophic growth is proposed to be maintained through restoration of the heterokaryotic state by continual anastomosis between adjacent homokaryotic hyphae. Since heterokaryosis is a prerequisite for parasexual recombination, we speculate that this mechanism is unlikely to play a major role in generating the VCG diversity found among Forl or other strains of F. oxysporum. [source] Comparative morphometrics of embryonic facial morphogenesis: Implications for cleft-lip etiologyTHE ANATOMICAL RECORD : ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY, Issue 1 2007Nathan M. Young Abstract Cleft lip (CL) with or without cleft palate (CL[P]) has a complex etiology but is thought to be due to either genetic or environmentally induced disruptions of developmental processes affecting the shape and size of the facial prominences (medial nasal, lateral nasal, and maxilla). Recent advances in landmark-based morphometrics enable a rigorous reanalysis of phenotypic shape variation associated with facial clefting. Here we use geometric morphometric (GM) tools to characterize embryonic shape variation in the midface and head of six strains of mice that are both cleft-liable (A, A/WySn, CL/Fr) and normal (BALB/cBy, C57BL, CD1). Data were comprised of two-dimensional landmarks taken from frontal and lateral photographs of embryos spanning the time period in which the facial prominences fuse (GD10-12). Results indicate that A/- strain mice, and particularly A/WySn, have overall smaller midfaces compared to other strains. The A/WySn strain also has significant differences in facial shape related to retarded development. Overall, CL/Fr strain mice are normal-sized, but tend to have undersized maxillary prominences that do not project anteriorly and have a small nasal contact area. These results suggest that the etiology of clefting differs in A/WySn and CL/Fr strains, with the former strain suffering disruptions to developmental processes affecting overall size (e.g., neural crest migration deficiencies and lower mitotic activity), while the latter strain has defects restricted to the shape and size of the maxilla. A combination of molecular experimentation and phenotypic analysis of shape is required to test these hypotheses further. Anat Rec, 290:123,139, 2007. © 2006 Wiley-Liss, Inc. [source] Characterization of R-body genetic determinants in Caedibacter caryophilus a symbiont of Paramecium caudatum: preliminary resultsTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 2 2005MARTINA SCHRALLHAMMER Refractile inclusion bodies, called R-bodies were observed within the cells of some bacterial strains. They are protein ribbons, which are typically coiled into cylindrical structures. They are produced by members of the genus Caedibacter, gram-negative rod-shaped endosymbionts of paramecia and e.g. the free-living bacteria Hydrogenophaga taeniospiralis and Acidovorax avenae. The phylogenetic relationship even between the members of the genus Caedibacter is quite low: C. taeniospiralis belongs to the Gammaproteobacteria and is related to Francisella tularensis, C. caryophilus is affiliated with the Alphaproteobacteria and clusters with the obligate endosymbiont Holospora. In the case of C. taeniospiralis 51, the genetic determinants of R-body synthesis are located on a plasmid, whereas in other strains like 7 and 562 it looks like phage particles are involved in their production. In the present study, we investigated C. caryophilus, endosymbiont of Paramecium caudatum. Separation of C. caryophilus cells was performed by PercollÔ density gradient centrifugation. The isolated DNA was separated by pulsed-field gel electrophoresis and it was possible to visualize several bands referring to one or more extrachromosomal elements. A small gene library of these extrachromosomal elements was constructed and we already identified transposition-related genes; interestingly, similar genes were reported also on the plasmid of C. taeniospiralis 51. [source] | ||||||||||||||||||||||||||||||||||