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1.
Ferro S Coppellotti O Roncucci G Ben Amor T Jori G 《Journal of applied microbiology》2006,101(1):206-212
AIMS: To develop alternative approaches for medical and environmental control of pathogenic Acanthamoeba spp. by means of photodynamic treatment with a tetracationic Zn(II)-phthalocyanine (RLP068). METHODS AND RESULTS: Incubation of cyst cultures with RLP068 for 1 h caused an accumulation of readily detectable concentrations of the phthalocyanine, even at doses as low as 0.5 micromol l(-1). RLP068 exhibited no dark toxicity towards cysts up to 5 micromol l(-1) concentration. A decrease of c. 50% in cyst survival in comparison with controls was measured upon incubation of the cysts with 0.5 micromol l(-1) RLP068, followed by exposure to light (600-700 nm) for 20 min at a fluence rate of 50 mW cm(-2) (60 J cm(-2)). After incubation with 3 and 5 micromol l(-1) RLP068 and irradiation, the cysts lost their excystment ability as early as day 5 and up to day 10, and were clearly damaged when observed under an interference contrast microscope. CONCLUSIONS: These data indicate the promising use of RLP068 in phototreatment of diseases caused by pathogenic amoebae and in initial disinfection of wastewaters. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and extensive photodamage may be induced in the highly resistant cystic stages by means of 600- to 700-nm light sources. 相似文献
2.
F.S. Martins L.C. Veloso R.M.E. Arantes J.R. Nicoli 《Letters in applied microbiology》2009,49(6):738-744
Aims: To compare the effects of five yeast probiotic formulations on viability, revival and washout kinetic in the digestive tract of mice, and the protection against an experimental infection with Salmonella enterica serovar Typhimurium.
Methods and Results: The number of viable cells in five commercial probiotic products codified as A, B, C and D ( Saccharomyces boulardii – lyophilized) and E ( Saccharomyces cerevisiae – aqueous suspension) was determined, as well as revival and washout kinetic in mouse intestine. Protective capacity was evaluated by survival rate and histopathology of liver and intestine of mice treated with each product and then challenged with Salm . Typhimurium.
Conclusions: Product A contained the highest number of viable cells and, fed to mice, gave the highest counts of viable yeasts and the longest persistence in faeces. Probably as a consequence, the highest survival and protection of intestinal and hepatic tissues were observed when product A was used for mouse treatment. Product E showed low counts in the formulation and was not recovered from mouse intestine.
Significance and Impact of the Study: Formulation (lyophilization or aqueous suspension) is an important factor for revival and survival of a probiotic product in vivo and consequently for its protective properties. 相似文献
Methods and Results: The number of viable cells in five commercial probiotic products codified as A, B, C and D ( Saccharomyces boulardii – lyophilized) and E ( Saccharomyces cerevisiae – aqueous suspension) was determined, as well as revival and washout kinetic in mouse intestine. Protective capacity was evaluated by survival rate and histopathology of liver and intestine of mice treated with each product and then challenged with Salm . Typhimurium.
Conclusions: Product A contained the highest number of viable cells and, fed to mice, gave the highest counts of viable yeasts and the longest persistence in faeces. Probably as a consequence, the highest survival and protection of intestinal and hepatic tissues were observed when product A was used for mouse treatment. Product E showed low counts in the formulation and was not recovered from mouse intestine.
Significance and Impact of the Study: Formulation (lyophilization or aqueous suspension) is an important factor for revival and survival of a probiotic product in vivo and consequently for its protective properties. 相似文献
3.
Chiti G Municchi M Paschetta V Nistri D Roncucci G 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2004,809(1):167-174
The development and validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of the tetracationic zinc(II) phthalocyanine derivative RLP068 in rabbit serum is described. The dodecadeuterated product (RLP068-D12) was used as co-eluting internal standard. RLP068 was isolated from serum samples by solid-phase extraction using weak cationic exchange cartridges (WCX). An oxidative derivatisation was used in order to simplify the peculiar HPLC and MS behaviour of the analyte and thus increasing sensitivity. Liquid Chromatography was carried out on a Polaris C18 Ether column (50 mm x 2.0 mm) with an isocratic run of 0.5% aqueous TFA/methanol. Detection was achieved by means of a Bruker Esquire 3000+ Ion Trap Mass Spectrometer equipped with an ESI source working in positive mode. A Multiple Reaction Monitoring method following the transitions 297.1 --> 282.1 for the analyte and 300.1 --> 282.1 + 285.1 for the internal standard was used. The analytical method was validated over the concentration range 2-65 ng/mL. lower limits of detection (LLOD) and quantification (LLOQ) were respectively 1 and 2 ng/mL. The method is innovative and applicable to pharmacokinetic studies. 相似文献
4.
5.
Aims: To design a rapid specific method for enumeration of viable Listeria spp. using the fluorescence in situ hybridization with filter cultivation (FISHFC) method.
Methods and Results: The probe, Lis-1400, was designed from the 23S rRNA region of the Listeria genome, and labelled with 5'-carboxy-tetramethyl-rhodamine- N - hydroxy-succinimide-ester. Fluorescence was observed for all Listeria species but not for any organisms from the other genera, suggesting Lis-1400 is highly specific for Listeria spp. For purposes of filter cultivation prior to hybridization, hydrophilic polypropylene membrane filters gave better contrast between fluorescing colonies and background fluorescence. This was because of a high S/N ratio (fluorescence intensity of each microcolony/fluorescence intensity of background noise) after FISH treatment. Results were achievable in 14 h using Lis-1400-aided FISHFC as compared with 4–7 days required for confirmation of Listeria spp. by conventional plate count methods. Moreover, viable Listeria counts in selected food samples showed no significant differences between Lis-1400-aided FISHFC and conventional methods.
Conclusions: The Lis-1400-aided FISHFC method is more efficient than conventional methods for enumeration of viable Listeria spp. in food samples.
Significance and Impact of the Study: For enumeration of Listeria spp., Lis-1400-aided FISHFC method is equally accurate yet faster than conventional plate count methods, and can be valuable in the control of listeriosis. 相似文献
Methods and Results: The probe, Lis-1400, was designed from the 23S rRNA region of the Listeria genome, and labelled with 5'-carboxy-tetramethyl-rhodamine- N - hydroxy-succinimide-ester. Fluorescence was observed for all Listeria species but not for any organisms from the other genera, suggesting Lis-1400 is highly specific for Listeria spp. For purposes of filter cultivation prior to hybridization, hydrophilic polypropylene membrane filters gave better contrast between fluorescing colonies and background fluorescence. This was because of a high S/N ratio (fluorescence intensity of each microcolony/fluorescence intensity of background noise) after FISH treatment. Results were achievable in 14 h using Lis-1400-aided FISHFC as compared with 4–7 days required for confirmation of Listeria spp. by conventional plate count methods. Moreover, viable Listeria counts in selected food samples showed no significant differences between Lis-1400-aided FISHFC and conventional methods.
Conclusions: The Lis-1400-aided FISHFC method is more efficient than conventional methods for enumeration of viable Listeria spp. in food samples.
Significance and Impact of the Study: For enumeration of Listeria spp., Lis-1400-aided FISHFC method is equally accurate yet faster than conventional plate count methods, and can be valuable in the control of listeriosis. 相似文献
6.
A. L. Rodriguez F. S. Stefani C. E. de Oliveira Praes¶ A. Piaceski¶ M. P. Oliveira† P. Martins† V. D. da Silva‡ C. Bonorino§ M. E. Bauer§ 《Cell proliferation》2009,42(4):562-567
Due to an increasing number of skin diseases as a result of exposure to ultraviolet (UV) radiation, it is necessary to evaluate the effectiveness of new skin care formulations with broad-spectrum sunscreens.
Objectives: This study aims to assess the status of nerve fibres in healthy human skin, to quantify effects of UV radiation on nerve endings, and to evaluate neuroprotective effects of new skin care formulations against UV exposure damage.
Methods: Samples were obtained from 34 female patients enrolled for plastic surgery and were immediately treated (10 min) with three emulsions: Cream 1, Cream 2 (placebo) and a sunscreen with sun protection factor 15 (SPF15). Control samples and those treated with the cream emulsions were exposed to UVA and UVB for 60 min. Nerve fibres were identified by immunofluorescence using a monoclonal antibody (anti-human CD56/NCAM). Cell damage was assessed by image analysis.
Results: Several cellular nervous structures were identified in the skin samples, including free nerve endings. UVA and UVB significantly decreased (40–60%) density of nerve endings in the control samples and those treated with placebo (Cream 2) or SPF15 (all P < 0.001). Cream 1 completely blocked effects of UV radiation on nerve endings ( P > 0.05 vs. control).
Conclusions: Quantification of cell damage induced by UV radiation provides useful information for identification of new skin care compounds with neuroprotective properties. 相似文献
Objectives: This study aims to assess the status of nerve fibres in healthy human skin, to quantify effects of UV radiation on nerve endings, and to evaluate neuroprotective effects of new skin care formulations against UV exposure damage.
Methods: Samples were obtained from 34 female patients enrolled for plastic surgery and were immediately treated (10 min) with three emulsions: Cream 1, Cream 2 (placebo) and a sunscreen with sun protection factor 15 (SPF15). Control samples and those treated with the cream emulsions were exposed to UVA and UVB for 60 min. Nerve fibres were identified by immunofluorescence using a monoclonal antibody (anti-human CD56/NCAM). Cell damage was assessed by image analysis.
Results: Several cellular nervous structures were identified in the skin samples, including free nerve endings. UVA and UVB significantly decreased (40–60%) density of nerve endings in the control samples and those treated with placebo (Cream 2) or SPF15 (all P < 0.001). Cream 1 completely blocked effects of UV radiation on nerve endings ( P > 0.05 vs. control).
Conclusions: Quantification of cell damage induced by UV radiation provides useful information for identification of new skin care compounds with neuroprotective properties. 相似文献
7.
S. Bouhdid J. Abrini A. Zhiri M.J. Espuny A. Manresa 《Journal of applied microbiology》2009,106(5):1558-1568
Aims: Evaluation of the cellular effects of Origanum compactum essential oil on Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213.
Methods and Results: The damage induced by O. compactum essential oil on these two strains has been studied using different techniques: plate count, potassium leakage, flow cytometry (FC) and transmission electron microscopy (TEM). The results showed that oil treatment led to reduction of cells viability and dissipated potassium ion gradients. Flow cytometric analysis showed that oil treatment promoted the accumulation of bis-oxonol and the membrane-impermeable nucleic acid stain propidium iodide (PI), indicating the loss of membrane potential and permeability. The ability to reduce 5-cyano-2,3-ditolyl tetrazolium chloride was inhibited. Unlike in Ps. aeruginosa , membrane potential and membrane permeability in Staph. aureus cells were affected by oil concentration and contact time. Finally, TEM showed various structural effects. Mesosome-like structures were seen in oil-treated Staph. aureus cells whereas in Ps. aeruginosa, coagulated cytoplasmic material and liberation of membrane vesicles were observed, and intracellular material was seen in the surrounding environment. Both FC and TEM revealed that the effects in Ps. aeruginosa were greater than in Staph. aureus .
Conclusions: Oregano essential oil induces membrane damage showed by the leakage of potassium and uptake of PI and bis-oxonol. Ultrastructural alterations and the loss of cell viability were observed.
Significance and Impact of the Study: Understanding the mode of antibacterial effect of the oil studied is of a great interest in it further application as natural preservative in food or pharmaceutical industries. 相似文献
Methods and Results: The damage induced by O. compactum essential oil on these two strains has been studied using different techniques: plate count, potassium leakage, flow cytometry (FC) and transmission electron microscopy (TEM). The results showed that oil treatment led to reduction of cells viability and dissipated potassium ion gradients. Flow cytometric analysis showed that oil treatment promoted the accumulation of bis-oxonol and the membrane-impermeable nucleic acid stain propidium iodide (PI), indicating the loss of membrane potential and permeability. The ability to reduce 5-cyano-2,3-ditolyl tetrazolium chloride was inhibited. Unlike in Ps. aeruginosa , membrane potential and membrane permeability in Staph. aureus cells were affected by oil concentration and contact time. Finally, TEM showed various structural effects. Mesosome-like structures were seen in oil-treated Staph. aureus cells whereas in Ps. aeruginosa, coagulated cytoplasmic material and liberation of membrane vesicles were observed, and intracellular material was seen in the surrounding environment. Both FC and TEM revealed that the effects in Ps. aeruginosa were greater than in Staph. aureus .
Conclusions: Oregano essential oil induces membrane damage showed by the leakage of potassium and uptake of PI and bis-oxonol. Ultrastructural alterations and the loss of cell viability were observed.
Significance and Impact of the Study: Understanding the mode of antibacterial effect of the oil studied is of a great interest in it further application as natural preservative in food or pharmaceutical industries. 相似文献
8.
Abstract. 1. Herbivory can induce resistance in a plant and the induced phenotype may be disfavoured by subsequent herbivores. Yet, as the distance between plants in a population increases, limited mobility may make a herbivore more likely to feed and oviposit on host plants in its immediate surroundings.
2. The present study tested whether a herbivore's preference and distribution across plants with different induced phenotypes was influenced by the spatial distribution of plants. A fragmented population of Solanum dulcamara plants was created. This consisted of discrete, spatially separated patches with different histories of damage, either herbivory from adult flea beetles ( Psylliodes affinis ), tortoise beetles ( Plagiometriona clavata ), or mechanical damage. Each patch was separated by 7 m and consisted of 12 plants that were spaced 30 cm apart. Then a fixed number of adult tortoise beetles were introduced to each patch, and movement and oviposition within and between spatially separate homogeneous patches (receiving one type of damage) were compared with movement and oviposition within heterogeneous patches (containing all three types of damage) over the growing season.
3. Flea beetle and tortoise beetle herbivory consistently induced different phytochemical responses in S. dulcamara (polyphenol oxidase and peroxidase), and adult tortoise beetles avoided oviposition on the flea beetle induced plants within heterogeneous patches. However, between homogeneous patches, plant phenotype did not influence oviposition. Colonisation by naturally occurring flea beetle adults followed a similar pattern.
4. These results suggest that the heterogeneity of plant phenotypes can influence herbivore choice and distribution at small but not large spatial scales. 相似文献
2. The present study tested whether a herbivore's preference and distribution across plants with different induced phenotypes was influenced by the spatial distribution of plants. A fragmented population of Solanum dulcamara plants was created. This consisted of discrete, spatially separated patches with different histories of damage, either herbivory from adult flea beetles ( Psylliodes affinis ), tortoise beetles ( Plagiometriona clavata ), or mechanical damage. Each patch was separated by 7 m and consisted of 12 plants that were spaced 30 cm apart. Then a fixed number of adult tortoise beetles were introduced to each patch, and movement and oviposition within and between spatially separate homogeneous patches (receiving one type of damage) were compared with movement and oviposition within heterogeneous patches (containing all three types of damage) over the growing season.
3. Flea beetle and tortoise beetle herbivory consistently induced different phytochemical responses in S. dulcamara (polyphenol oxidase and peroxidase), and adult tortoise beetles avoided oviposition on the flea beetle induced plants within heterogeneous patches. However, between homogeneous patches, plant phenotype did not influence oviposition. Colonisation by naturally occurring flea beetle adults followed a similar pattern.
4. These results suggest that the heterogeneity of plant phenotypes can influence herbivore choice and distribution at small but not large spatial scales. 相似文献
9.
Aims: To investigate the antibacterial mechanism of carvacrol and thymol against Escherichia coli.
Methods and Results: The time-kill curve results showed that carvacrol and thymol at 200 mg l−1 could inhibit the growth of E. coli . Flow cytometry and fluorescent dyes were used to explore the effect of two components on membrane permeability and membrane potential. In membrane permeability experiment, the mean fluorescence intensity of cells treated with 200 mg l−1 carvacrol or thymol were lower than nonexposed cells. The ratio of red to green fluorescence intensity of DiOC2 (3) reflected the change of membrane potential. Carvacrol and thymol at 200 mg l−1 caused the ratio of red/green decreasing from 0·42 of control to 0·08 and 0·07, respectively.
Conclusions: Carvacrol and thymol had desired antimicrobial effect on E. coli . The antibacterial effects were attributed to their ability to permeabilize and depolarize the cytoplasmic membrane.
Significance and Impact of the Study: This study showed the potential use of flow cytometry as a suitable method to investigate the mode of antibacterial action of essential oil components. 相似文献
Methods and Results: The time-kill curve results showed that carvacrol and thymol at 200 mg l
Conclusions: Carvacrol and thymol had desired antimicrobial effect on E. coli . The antibacterial effects were attributed to their ability to permeabilize and depolarize the cytoplasmic membrane.
Significance and Impact of the Study: This study showed the potential use of flow cytometry as a suitable method to investigate the mode of antibacterial action of essential oil components. 相似文献
10.
Aims: In vitro experiments were undertaken to evaluate biocide formulations commonly used in cooling water systems against protozoa previously isolated from cooling towers. The investigations evaluated the efficacy of these formulations against amoebic cysts and trophozoites.
Methods and Results: Laboratory challenges against protozoa isolated from cooling towers using chlorine, bromine and isothiazolinone biocides showed that all were effective after 4 h. The presence of molybdate and organic phosphates resulted in longer kill times for bromine and isothiazolinones. All treatments resulted in no detectable viable protozoa after 4 h of exposure.
Conclusions: The chemical disinfection of planktonic protozoa in cooling water systems is strongly influenced by the residence time of the formulation and less so by its active constituent. Bromine and isothiazolinone formulations may require higher dosage of concentrations than currently practiced if used in conjunction with molybdate- and phosphate-based scale/corrosion inhibitors.
Significance and Impact of the Study: Cooling water systems are complex microbial ecosystems in which predator–prey relationships play a key role in the dissemination of Legionella . This study demonstrated that at recommended dosing concentrations, biocides had species-specific effects on environmental isolates of amoebae that may act as reservoirs for Legionella multiplication in cooling water systems. 相似文献
Methods and Results: Laboratory challenges against protozoa isolated from cooling towers using chlorine, bromine and isothiazolinone biocides showed that all were effective after 4 h. The presence of molybdate and organic phosphates resulted in longer kill times for bromine and isothiazolinones. All treatments resulted in no detectable viable protozoa after 4 h of exposure.
Conclusions: The chemical disinfection of planktonic protozoa in cooling water systems is strongly influenced by the residence time of the formulation and less so by its active constituent. Bromine and isothiazolinone formulations may require higher dosage of concentrations than currently practiced if used in conjunction with molybdate- and phosphate-based scale/corrosion inhibitors.
Significance and Impact of the Study: Cooling water systems are complex microbial ecosystems in which predator–prey relationships play a key role in the dissemination of Legionella . This study demonstrated that at recommended dosing concentrations, biocides had species-specific effects on environmental isolates of amoebae that may act as reservoirs for Legionella multiplication in cooling water systems. 相似文献
11.
M.R. Van Leeuwen E.A. Golovina J. Dijksterhuis 《Journal of applied microbiology》2009,106(6):1908-1918
Aims: To investigate the differences in membrane permeability and the effect on endocytosis of the polyene antimycotics nystatin, filipin and natamycin on germinating fungal conidia.
Methods and Results: The model system was Penicillium discolor , a food spoilage fungus. Filipin resulted in permeabilization of germinating conidia for the fluorescent probes TOTO-1 and FM4-64, but not for ferricyanide ions. Nystatin caused influx of all these compounds while natamycin did not. Untreated germinating conidia internalize the endocytic marker FM4-64. Pretreatment of germinating conidia with natamycin showed a dose and time dependent inhibition of endocytosis as judged by the lack of formation of early endosomal compartments.
Conclusions: The results obtained from this study indicated that, unlike nystatin and filipin, natamycin is unable to permeabilize germinating conidia, but interferes with endocytosis in a dose and time dependent manner.
Significance and Impact of the Study: Natamycin acts via a different mode of action than other polyene antimycotics. These results offer useful information for new strategies to prevent fungal spoilage on food products and infection on agricultural crops. For laboratory use, natamycin can be used as a specific inhibitor of early endocytosis in fungal cells. 相似文献
Methods and Results: The model system was Penicillium discolor , a food spoilage fungus. Filipin resulted in permeabilization of germinating conidia for the fluorescent probes TOTO-1 and FM4-64, but not for ferricyanide ions. Nystatin caused influx of all these compounds while natamycin did not. Untreated germinating conidia internalize the endocytic marker FM4-64. Pretreatment of germinating conidia with natamycin showed a dose and time dependent inhibition of endocytosis as judged by the lack of formation of early endosomal compartments.
Conclusions: The results obtained from this study indicated that, unlike nystatin and filipin, natamycin is unable to permeabilize germinating conidia, but interferes with endocytosis in a dose and time dependent manner.
Significance and Impact of the Study: Natamycin acts via a different mode of action than other polyene antimycotics. These results offer useful information for new strategies to prevent fungal spoilage on food products and infection on agricultural crops. For laboratory use, natamycin can be used as a specific inhibitor of early endocytosis in fungal cells. 相似文献
12.
C. Baker-Austin J. Morris J.A. Lowther R. Rangdale D.N. Lees 《Letters in applied microbiology》2009,49(4):529-532
Aims: To develop a quick, easy-to-use, robust and sensitive multiplex PCR assay to detect common sources of agricultural faecal contamination using a combination of bacterial and eukaryote-specific PCR targets.
Method and Results: A novel multiplex PCR method was developed that utilizes primers specific for a conserved region of the eukaryote cytochrome-B gene as well as a universal 16S rRNA and the E. coli -specific uidA gene. This multiplex PCR assay was capable of identifying faecal amendments from pig, sheep, cow and goat sources in 24/30 (80%) of amended water samples.
Conclusions: The method was capable of accurately identifying common agricultural sources.
Significance and Impact of the study: The procedure described here is simple, rapid (<5 h) and can be used as a first step in microbial source tracking studies, particularly where agricultural faecal contamination is suspected. 相似文献
Method and Results: A novel multiplex PCR method was developed that utilizes primers specific for a conserved region of the eukaryote cytochrome-B gene as well as a universal 16S rRNA and the E. coli -specific uidA gene. This multiplex PCR assay was capable of identifying faecal amendments from pig, sheep, cow and goat sources in 24/30 (80%) of amended water samples.
Conclusions: The method was capable of accurately identifying common agricultural sources.
Significance and Impact of the study: The procedure described here is simple, rapid (<5 h) and can be used as a first step in microbial source tracking studies, particularly where agricultural faecal contamination is suspected. 相似文献
13.
Marti M Regös A Li Y Schraner EM Wild P Müller N Knopf LG Hehl AB 《The Journal of biological chemistry》2003,278(27):24837-24848
14.
Abstract. 1. Interactions among predators may influence the total efficiency of a predator complex. The effect of intra- and interspecific interactions of the generalist predators Orthotylus marginalis (Heteroptera: Miridae) and Anthocoris nemorum (Heteroptera: Anthocoridae) was investigated in a laboratory experiment. Outcomes of the interactions were determined by comparing predation rates on eggs and larvae of the blue willow beetle Phratora vulgatissima of single individuals with those of two individuals of the same or different species.
2. A non-additive, antagonistic effect on predation rates due to intraspecific interactions was found between individuals of A. nemorum . No such effect was found in O. marginalis . These results are as expected as a consequence of differences in behaviour of the two predator species: A. nemorum is a much more active and mobile predator than O. marginalis .
3. Contrary to expectation, interspecific interactions between A. nemorum and O. marginalis did not affect the total predation rate.
4. An observation from the field corroborated the results obtained in the laboratory study; there was no negative relationship between the densities of the two predator species, indicating that the two species do not interact negatively in the field at their natural densities.
5. It is concluded that the additive effect of multiple predator species is of potential value in biological control. 相似文献
2. A non-additive, antagonistic effect on predation rates due to intraspecific interactions was found between individuals of A. nemorum . No such effect was found in O. marginalis . These results are as expected as a consequence of differences in behaviour of the two predator species: A. nemorum is a much more active and mobile predator than O. marginalis .
3. Contrary to expectation, interspecific interactions between A. nemorum and O. marginalis did not affect the total predation rate.
4. An observation from the field corroborated the results obtained in the laboratory study; there was no negative relationship between the densities of the two predator species, indicating that the two species do not interact negatively in the field at their natural densities.
5. It is concluded that the additive effect of multiple predator species is of potential value in biological control. 相似文献
15.
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). 相似文献
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 T
Conclusions: Bifidobacterium spp. was the population with the lowest persistence, showing seasonal differences in T
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). 相似文献
16.
Use of selected enterococci and Rhizopus oryzae proteases to hydrolyse wheat proteins responsible for celiac disease 总被引:1,自引:0,他引:1
S. M'hir C.G. Rizzello R. Di Cagno A. Cassone M. Hamdi 《Journal of applied microbiology》2009,106(2):421-431
Aims: This work aimed at using a pool of selected enterococci and fungal proteases to hydrolyse wheat gluten during long-time fermentation.
Methods and Results: A liquid dough made with wheat flour (20% w/w) was fermented with three Enterococcus strains (dough A) or with the combination of enterococci and Rhizopus oryzae proteases (dough B). After 48 h of fermentation, dough A and B had a concentration of water-soluble peptides approximately threefold higher than the chemically acidified dough (CAD), used as the control. The same was found for the concentration of free amino acids, being higher in dough B with respect to dough A. SDS-PAGE analysis showed that albumin and glutenin fractions were partially hydrolysed, while gliadins almost disappeared in dough A and B, as confirmed by two-dimensional electrophoresis, RP-HPLC and R5-ELISA analyses.
Conclusions: The combined use of enterococci and fungal proteases showed a decrease of the gluten concentration of more than 98% during long-time fermentation.
Significance and Impact of the Study: The use of the mixture of selected enterococci and R. oryzae proteases should be considered as a potential tool to decrease gluten concentration in foods. 相似文献
Methods and Results: A liquid dough made with wheat flour (20% w/w) was fermented with three Enterococcus strains (dough A) or with the combination of enterococci and Rhizopus oryzae proteases (dough B). After 48 h of fermentation, dough A and B had a concentration of water-soluble peptides approximately threefold higher than the chemically acidified dough (CAD), used as the control. The same was found for the concentration of free amino acids, being higher in dough B with respect to dough A. SDS-PAGE analysis showed that albumin and glutenin fractions were partially hydrolysed, while gliadins almost disappeared in dough A and B, as confirmed by two-dimensional electrophoresis, RP-HPLC and R5-ELISA analyses.
Conclusions: The combined use of enterococci and fungal proteases showed a decrease of the gluten concentration of more than 98% during long-time fermentation.
Significance and Impact of the Study: The use of the mixture of selected enterococci and R. oryzae proteases should be considered as a potential tool to decrease gluten concentration in foods. 相似文献
17.
Frumerie C Sylwan L Helleday T Yu A Haggård-Ljungquist E 《Journal of applied microbiology》2008,105(1):290-299
Aims: To investigate if the site-specific tyrosine integrase (Int) from phage P2 has features that would make it interesting for use of gene transfer into eukaryotic cells. These include the possibility of promoting recombination with a nonphage sequence, abolishing the requirement for the bacterial DNA-binding and -bending protein integration host factor (IHF), and localization to the nucleus of eukaryotic cells.
Methods and Results: We show that the Int protein catalyzes site-specific recombination using a human sequence in Escherichia coli and in vitro although not as efficiently as with the wild-type bacterial sequence, and that insertion of high mobility group recognition boxes in the phage attachment site substrate abolish the requirement of IHF and allows efficient recombination in vitro in a eukaryotic cell extract. Furthermore, we show by fluorescence that the Int protein contains a functional intrinsic nuclear localization signal, localizing it to the nucleus in both HeLa and 293 cells.
Conclusions: We conclude that P2 Int may be a potential tool for site-specific integration of genes into the human chromosome.
Significance and Impact of the Study: The study implies the possibility of using multiple prokaryotic Int proteins with different specific integration sites in human cells for future gene therapy programmes. 相似文献
Methods and Results: We show that the Int protein catalyzes site-specific recombination using a human sequence in Escherichia coli and in vitro although not as efficiently as with the wild-type bacterial sequence, and that insertion of high mobility group recognition boxes in the phage attachment site substrate abolish the requirement of IHF and allows efficient recombination in vitro in a eukaryotic cell extract. Furthermore, we show by fluorescence that the Int protein contains a functional intrinsic nuclear localization signal, localizing it to the nucleus in both HeLa and 293 cells.
Conclusions: We conclude that P2 Int may be a potential tool for site-specific integration of genes into the human chromosome.
Significance and Impact of the Study: The study implies the possibility of using multiple prokaryotic Int proteins with different specific integration sites in human cells for future gene therapy programmes. 相似文献
18.
Postharvest biocontrol of Alternaria alternata in Chinese winter jujube by Rhodosporidium paludigenum 总被引:1,自引:0,他引:1
Y. Wang T. Yu Y. Li D. Cai X. Liu H. Lu X.D. Zheng 《Journal of applied microbiology》2009,107(5):1492-1498
Aims: This study was conducted to measure the efficacy of the marine antagonist Rhodosporidium paludigenum in the suppression of postharvest decay of Chinese winter jujube caused by Alternaria alternata and to explore the possible mode of action involved.
Methods and Results: The efficacy of controlling postharvest diseases by R. paludigenum was examined. Rapid yeast colonization of wounds was observed during the first 48 h at 25°C. The yeast at 1 × 108 cells ml−1 of washed cells suspension provided better control of A. alternata than any other treatment. The concentration of the antagonist had significant effects on biocontrol effectiveness: as the concentration of R. paludigenum was increased, the disease incidence decreased. Meanwhile, R. paludigenum significantly inhibited the natural development of decay and did not damage fruit quality parameters including lightness values, hue angle, firmness, soluble solids, ascorbic acid and titratable acidity in 21 days' storage at 25°C.
Conclusions: Rhodosporidium paludigenum was effective in controlling postharvest decay of Chinese winter jujube and did not impair fruit quality parameters.
Significance and Impact of the Study: Rhodosporidium paludigenum can be used as a nonchemical agent in postharvest biological control of Chinese winter jujube. 相似文献
Methods and Results: The efficacy of controlling postharvest diseases by R. paludigenum was examined. Rapid yeast colonization of wounds was observed during the first 48 h at 25°C. The yeast at 1 × 10
Conclusions: Rhodosporidium paludigenum was effective in controlling postharvest decay of Chinese winter jujube and did not impair fruit quality parameters.
Significance and Impact of the Study: Rhodosporidium paludigenum can be used as a nonchemical agent in postharvest biological control of Chinese winter jujube. 相似文献
19.
Antibacterial activities of zinc oxide nanoparticles against Escherichia coli O157:H7 总被引:2,自引:0,他引:2
Y. Liu L. He A. Mustapha H. Li Z.Q. Hu M. Lin 《Journal of applied microbiology》2009,107(4):1193-1201
Aims: To investigate antibacterial activities of zinc oxide nanoparticles (ZnO NP) and their mode of action against an important foodborne pathogen, Escherichia coli O157:H7.
Methods and Results: ZnO NP with sizes of 70 nm and concentrations of 0, 3, 6 and 12 mmol l−1 and NP-free solutions were used in antimicrobial tests against E. coli O157:H7. ZnO NP showed increasing inhibitory effects on the growth of E. coli O157:H7 as the concentrations of ZnO NP increased. A complete inhibition of microbial growth was achieved at the concentration level of 12 mmol l−1 or higher. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Raman spectroscopy were used to characterize the changes of morphology and cellular compositions of bacterial cells treated with ZnO NP and study the mode of action of ZnO NP against E. coli O157:H7. The intensity of lipid and protein bands in the Raman spectra of bacterial cells increased after exposure to ZnO NP, while no significant changes in nucleic acid bands were observed.
Conclusions: ZnO NP were found to have antibacterial activity against E. coli O157:H7. The inhibitory effects increase as the concentration of ZnO NP increased. Results indicate that ZnO NP may distort and damage bacterial cell membrane, resulting in a leakage of intracellular contents and eventually the death of bacterial cells.
Significance and Impact of the Study: These results suggest that ZnO NP could potentially be used as an effective antibacterial agent to protect agricultural and food safety. 相似文献
Methods and Results: ZnO NP with sizes of 70 nm and concentrations of 0, 3, 6 and 12 mmol l
Conclusions: ZnO NP were found to have antibacterial activity against E. coli O157:H7. The inhibitory effects increase as the concentration of ZnO NP increased. Results indicate that ZnO NP may distort and damage bacterial cell membrane, resulting in a leakage of intracellular contents and eventually the death of bacterial cells.
Significance and Impact of the Study: These results suggest that ZnO NP could potentially be used as an effective antibacterial agent to protect agricultural and food safety. 相似文献
20.
C.M. Yang M.F. Lin P.C. Liao H.W. Yeh B.V. Chang T.K. Tang C. Cheng C.H. Sung M.L. Liou 《Letters in applied microbiology》2009,48(5):560-565
Aims: To compare bacterial populations and antimicrobial resistance patterns between clinical and sewage isolates from a regional hospital in northern Taiwan. The dissemination of antibiotic-resistant bacteria from hospital compartments to the hospital sewage treatment plant was examined.
Methods and Results: A total of 1020 clinical isolates and 435 sewage isolates were collected between July and September 2005. The percentages of Gram-negative bacteria from the clinical and sewage isolates were 87·2% and 91·0%, respectively ( P = 0·033). Escherichia coli were the leading bacterial isolates in both groups. Antimicrobial susceptibility testing showed a significant difference ( P < 0·001) in resistance to ampicillin (85·6% vs 94·1%), ampicillin/sulbactam (31·7% vs 55·4%), cefazolin (29·2% vs 71·5%) and cefuroxime (20·7% vs 61·9%) between clinical and sewage coliform isolates, respectively.
Conclusions: The sewage isolates had higher antimicrobial resistance rates than the clinical isolates from the same hospital.
Significance and Impact of the Study: The low efficacy of the hospital sewage treatment may contribute to the dissemination of multidrug resistant bacteria from this hospital compartments to the environment. Practices which limit the disposal of antimicrobial agents into the wastewater system may be the possible measure to prevent the selection of multidrug-resistant bacteria from sewage treatment plants. 相似文献
Methods and Results: A total of 1020 clinical isolates and 435 sewage isolates were collected between July and September 2005. The percentages of Gram-negative bacteria from the clinical and sewage isolates were 87·2% and 91·0%, respectively ( P = 0·033). Escherichia coli were the leading bacterial isolates in both groups. Antimicrobial susceptibility testing showed a significant difference ( P < 0·001) in resistance to ampicillin (85·6% vs 94·1%), ampicillin/sulbactam (31·7% vs 55·4%), cefazolin (29·2% vs 71·5%) and cefuroxime (20·7% vs 61·9%) between clinical and sewage coliform isolates, respectively.
Conclusions: The sewage isolates had higher antimicrobial resistance rates than the clinical isolates from the same hospital.
Significance and Impact of the Study: The low efficacy of the hospital sewage treatment may contribute to the dissemination of multidrug resistant bacteria from this hospital compartments to the environment. Practices which limit the disposal of antimicrobial agents into the wastewater system may be the possible measure to prevent the selection of multidrug-resistant bacteria from sewage treatment plants. 相似文献