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1.

Aims

To determine the fate of Escherichia coli on vegetables that were processed through commercial wash treatments and stored under simulated retail conditions at 4°C or wholesale at fluctuating ambient temperatures (0–25°C, dependent on season).

Methods and Results

Bovine slurry that was naturally contaminated with E. coli O145 was applied without dilution or diluted 1:10 using borehole water to growing potatoes, leeks or carrots. Manure was applied 1 week prior to harvest to simulate a near‐harvest contamination event by manure deposition or an application of contaminated water to simulate a flooding event or irrigation from a contaminated water source. At harvest, crops were contaminated at up to 2 log cfu g?1. Washing transferred E. coli into the water of a flotation tank used for potato washing and did not completely remove all traces of contamination from the crop. Manure‐contaminated potatoes were observed to contain 0·72 cfu E. coli O145 g?1 after processing and retail storage. Manure‐contaminated leeks harboured 0·73–1·55 cfu E. coli O145 g?1 after washing and storage. There was no cross‐contamination when leeks were spray washed. Washing in an abrasive drum resulted in less than perfect decontamination for manure‐contaminated carrots. There were five post‐distribution isolations from carrots irrigated with contaminated water 24 h prior to harvest.

Conclusions

Standard commercial washing and distribution conditions may be insufficient to reliably control human pathogenic E. coli on fresh produce.

Significance and Impact

Previous speculation that the cause of a UK foodborne disease outbreak was soil from imperfectly cleaned vegetables is plausible.  相似文献   

2.

Background

Shiga-toxin producing Escherichia coli (STEC) have emerged as important foodborne pathogens, among which seven serogroups (O26, O45, O103, O111, O121, O145, O157) are most frequently implicated in human infection. The aim was to determine if a light scattering sensor can be used to rapidly identify the colonies of STEC serogroups on selective agar plates.

Methodology/Principal Findings

Initially, a total of 37 STEC strains representing seven serovars were grown on four different selective agar media, including sorbitol MacConkey (SMAC), Rainbow Agar O157, BBL CHROMagarO157, and R&F E. coli O157:H7, as well as nonselective Brain Heart Infusion agar. The colonies were scanned by an automated light scattering sensor, known as BARDOT (BActerial Rapid Detection using Optical scattering Technology), to acquire scatter patterns of STEC serogroups, and the scatter patterns were analyzed using an image classifier. Among all of the selective media tested, both SMAC and Rainbow provided the best differentiation results allowing multi-class classification of all serovars with an average accuracy of more than 90% after 10–12 h of growth, even though the colony appearance was indistinguishable at that early stage of growth. SMAC was chosen for exhaustive scatter image library development, and 36 additional strains of O157:H7 and 11 non-O157 serovars were examined, with each serogroup producing unique differential scatter patterns. Colony scatter images were also tested with samples derived from pure and mixed cultures, as well as experimentally inoculated food samples. BARDOT accurately detected O157 and O26 serovars from a mixed culture and also from inoculated lettuce and ground beef (10-h broth enrichment +12-h on-plate incubation) in the presence of natural background microbiota in less than 24 h.

Conclusions

BARDOT could potentially be used as a screening tool during isolation of the most important STEC serovars on selective agar plates from food samples in less than 24 h.  相似文献   

3.

Aims

Integrated weed management, which allows reducing the reliance of cropping systems on herbicides, is based on the use of specific combinations of innovative agricultural practices. However the impact of the introduction of these practices in cropping systems may influence soil functioning such as biogeochemical cycling. Here, we investigated N2O emissions and the abundances of N-cycling microorganisms in 11-year old cropping systems (i.e. conventional reference and integrated weed management) in order to estimate the environmental side-effects of long-term integrated weed management.

Methods

N2O emissions were continuously measured using automated chambers coupled with infrared analysers. Abundances of ammonia oxidizers and denitrifiers together with total bacteria and archaea were determined monthly from 0 to 10 and 10–30 cm soil layer samples by quantitative Polymerase Chain Reaction (qPCR). The relationship between N2O emissions and microbial abundances during the study were investigated as were their relationships with soil physicochemical parameters and climatic conditions.

Results

Over 7 months, the system with integrated weed management emitted significantly more N2O with cumulated measured emissions of 240 and 544 g N-N2O ha?1 for conventional and integrated systems, respectively. Abundances of microbial guilds varied slightly between systems, although ammonia-oxidizing bacteria were more abundant in the reference system (1.7 106 gene copies g?1 dry weight soil) compared to the integrated system (1.0 106 gene copies g?1 dry weight soil). These differences revealed both the long-term modification of soil biogeochemical background and the functioning of microbial processes due to 11 years of alternative field management, and the short-term impacts of the agricultural practices introduced as part of weed management during the cropping year.

Conclusions

The abundances of the different microbial communities involved in N cycling and the intensity of N2O emissions were not related, punctual high N2O emissions being more dependent on favourable soil conditions for nitrifying and denitrifying activities. Future studies will be performed to check these findings for other pedoclimatic conditions and to examine the impact of such cropping systems.  相似文献   

4.

Background and aims

Elevated atmospheric CO2 (eCO2) and tropospheric O3 (eO3) can alter soil microbial processes, including those underlying N2O emissions, as an indirect result of changes in plant inputs. In this study, effects of eCO2 and eO3 on sources of N2O in a soybean (Glycine max (L.) Merr.) agroecosystem in Illinois (SoyFACE) were investigated. We hypothesized that increases in available C and anaerobic microhabitat under eCO2 would stimulate N2O emissions, with a proportionally larger increase in denitrification derived N2O (N2OD) compared to nitrification plus nitrifier denitrification derived N2O (N2ON+ND). We expected opposite effects under eO3.

Methods

Isotopically labeled 15NH 4 14 NO3 and 14NH 4 15 NO3 were used to evaluate mineral N transformations, N2OD, and N2ON+ND in a 12-day incubation experiment.

Results

We observed minimal effects of eCO2 and eO3 on N2O emissions, movement of 15?N through mineral N pools, soil moisture content and C availability. Possibly, altered C and N inputs by eCO2 and eO3 were small relative to the high soil organic C content and N-inputs via biological N2-fixation, minimizing potential effects of eCO2 and eO3 on N-cycling.

Conclusion

We conclude that eCO2 and eO3 did not affect N2O emissions in the short term. However, it remains to be tested whether N2O emissions in SoyFACE will be unaltered by eCO2 and eO3 on a larger temporal scale under field conditions.  相似文献   

5.

Purpose  

Recent literature has highlighted a renewed debate amongst the scientific community about the relevance of nitrous oxide (N2O) emissions to future ozone layer management. This raises the question as to whether the life cycle assessment (LCA) community should also consider incorporating N2O into its ozone depletion models. This discussion summarises a preliminary investigation into the justification for doing so.  相似文献   

6.

Introduction

Quantitative proteomics using tandem mass spectrometry is an attractive approach for identification of potential cancer biomarkers. Fractionation of complex tissue samples into subproteomes prior to mass spectrometric analyses increases the likelihood of identifying cancer-specific proteins that might be present in low abundance. In this regard, glycosylated proteins are an interesting class of proteins that are already established as biomarkers for several cancers.

Materials and Methods

In this study, we carried out proteomic profiling of tumor and adjacent non-cancer liver tissues from hepatocellular carcinoma (HCC) patients. Glycoprotein enrichment from liver samples using lectin affinity chromatography and subsequent 18O/16O labeling of peptides allowed us to obtain relative abundance levels of lectin-bound proteins. As a complementary approach, we also examined the relative expression of proteins in HCC without glycoprotein enrichment. Lectin affinity enrichment was found to be advantageous to quantitate several interesting proteins, which were not detected in the whole proteome screening approach. We identified and quantitated over 200 proteins from the lectin-based approach. Interesting among these were fetuin, cysteine-rich protein 1, serpin peptidase inhibitor, leucine-rich alpha-2-glycoprotein 1, melanoma cell adhesion molecule, and heparan sulfate proteoglycan-2. Using lectin affinity followed by PNGase F digestion coupled to 18O labeling, we identified 34 glycosylation sites with consensus sequence N-X-T/S. Western blotting and immunohistochemical staining were carried out for several proteins to confirm mass spectrometry results.

Conclusion

This study indicates that quantitative proteomic profiling of tumor tissue versus non-cancerous tissue is a promising approach for the identification of potential biomarkers for HCC.  相似文献   

7.
8.

Background and Aims

Tree species composition shifts can alter soil CO2 and N2O effluxes. We quantified the soil CO2 and N2O efflux rates and temperature sensitivity from Pyrenean oak, Scots pine and mixed stands in Central Spain to assess the effects of a potential expansion of oak forests.

Methods

Soil CO2 and N2O effluxes were measured from topsoil samples by lab incubation from 5 to 25 °C. Soil microbial biomass and community composition were assessed.

Results

Pine stands showed highest soil CO2 efflux, followed by mixed and oak forests (up to 277, 245 and 145 mg CO2-C m?2 h?1, respectively). Despite contrasting soil microbial community composition (more fungi and less actinomycetes in pine plots), carbon decomposability and temperature sensitivity of the soil CO2 efflux remain constant among tree species. Soil N2O efflux rates and its temperature sensitivity was markedly higher in oak stands than in pine stands (70 vs. 27 μg N2O-N m?2 h?1, Q10, 4.5 vs. 2.5).

Conclusions

Conversion of pine to oak forests in the region will likely decrease soil CO2 effluxes due to decreasing SOC contents on the long run and will likely enhance soil N2O effluxes. Our results present only a seasonal snapshot and need to be confirmed in the field.  相似文献   

9.

Background and aims

The direct measurement of denitrification dynamics and its product fractions is important for parameterizing process-oriented model(s) for nitrogen cycling in various soils. The aims of this study are to a) directly measure the denitrification potential and the fractions of nitrogenous gases as products of the process in laboratory, b) investigate the effects of the nitrate (NO 3 ? ) concentration on emissions of denitrification gases, and c) test the hypothesis that denitrification can be a major pathway of nitrous oxide (N2O) and nitric oxide (NO) production in calcic cambisols under conditions of simultaneously sufficient supplies of carbon and nitrogen substrates and anaerobiosis as to be found to occur commonly in agricultural lands.

Methods

Using the helium atmosphere (with or without oxygen) gas-flow-soil-core technique in laboratory, we directly measured the denitrification potential of a silt clay calcic cambisol and the production of nitrogen gas (N2), N2O and NO during denitrification under the conditions of seven levels of NO 3 ? concentrations (ranging from 10 to 250 mg N kg?1 dry soil) and an almost constant initial dissolved organic carbon concentration (300 mg C kg?1 dry soil).

Results

Almost all the soil NO 3 ? was consumed during anaerobic incubation, with 80–88 % of the consumed NO 3 ? recovered by measuring nitrogenous gases. The results showed that the increases in initial NO 3 ? concentrations significantly enhanced the denitrification potential and the emissions of N2 and N2O as products of this process. Despite the wide range of initial NO 3 ? concentrations, the ratios of N2, N2O and NO products to denitrification potential showed much narrower ranges of 51–78 % for N2, 14–36 % for N2O and 5–22 % for NO.

Conclusions

These results well support the above hypothesis and provide some parameters for simulating effects of variable soil NO 3 ? concentrations on denitrification process as needed for biogeochemical models.  相似文献   

10.

Background

An aptamer based biosensor (aptasensor) was developed and evaluated for rapid colorimetric detection of Escherichia coli (E. coli) O157:H7.

Methodology/Principal Findings

The aptasensor was assembled by modifying the truncated lipopolysaccharides (LPS)-binding aptamer on the surface of nanoscale polydiacetylene (PDA) vesicle using peptide bonding between the carboxyl group of the vesicle and the amine group of the aptamer. Molecular recognition between E. coli O157:H7 and aptamer at the interface of the vesicle lead to blue-red transition of PDA which was readily visible to the naked eyes and could be quantified by colorimetric responses (CR). Confocal laser scanning microscope (CLSM) and transmission electron microscopy (TEM) was used to confirm the specific interactions between the truncated aptamer and E. coli O157:H7. The aptasensor could detect cellular concentrations in a range of 104∼ 108 colony-forming units (CFU)/ml within 2 hours and its specificity was 100% for detection of E. coli O157:H7. Compared with the standard culture method, the correspondent rate was 98.5% for the detection of E. coli O157:H7 on 203 clinical fecal specimens with our aptasensor.

Conclusions

The new aptasensor represents a significant advancement in detection capabilities based on the combination of nucleic acid aptamer with PDA vesicle, and offers a specific and convenient screening method for the detection of pathogenic bacteria. This technic could also be applied in areas from clinical analysis to biological terrorism defense, especially in low-resource settings.  相似文献   

11.

Background and aims

Continuous vegetable cultivation in greenhouses can easily induce soil degradation, which considerably affects the development of sustainable vegetable production. Recently, the reductive soil disinfestation (RSD) is widely used as an alternative to chemical soil disinfestations to improve degraded greenhouse vegetable soils. Considering the importance of nitrogen (N) for plant growth and environment effect, the internal N transformation processes and rates should be well investigated in degraded vegetable soils treated by RSD, but few works have been undertaken.

Methods

Three RSD-treated and three untreated degraded vegetable soils were chosen and a 15?N tracing incubation experiment differentially labeled with 15NH4NO3 or NH4 15NO3 was conducted at 25 °C under 50 % water holding capacity (WHC) for 96 h. Soil gross N transformation rates were calculated using a 15?N tracing model combined with Markov Chain Monte Carlo Metropolis algorithm (Müller et al. 2007), while the emissions of N2O and NO were also measured.

Results

RSD could significantly enhance the soil microbial NH4 + immobilization rate, the heterotrophic and autotrophic nitrification rates, and the NO3 ? turnover time. The ratio of heterotrophic nitrification to total inorganic N supply rate (mineralization + heterotrophic nitrification) increased greatly from 5.4 % in untreated vegetable soil to 56.1 % in treated vegetable soil. In addition, low release potential of NO and N2O was observed in RSD-treated vegetable soil, due to the decrease in the NO and N2O product ratios from heterotrophic and autotrophic nitrifications. These significant differences in gross N transformation rates, the supply processes and capacity of inorganic N, and the NO and N2O emissions between untreated and treated vegetable soils could be explained by the elimination of accumulated NO3 ?, increased pH, and decreased electrical conductivity (EC) caused by RSD. Noticeably, the NO3 ? consumption rates were still significantly lower than the NO3 ? production rates in RSD-treated vegetable soil.

Conclusions

Except for improving soil chemical properties, RSD could significantly alter the supply processes of inorganic N and reduce the release potential of N2O and NO in RSD-treated degraded vegetable soil. In order to retard the re-occurrence of NO3 ? accumulation, acidification and salinization and to promote the long-term productivity of greenhouse vegetable fields, the rational use of N fertilizer should be paid great attention to farmers in vegetable cultivation.  相似文献   

12.

Objective

We identified a poor clinical response to treatment of cholera with a single 1 g dose of ciprofloxacin, a standard treatment for cholera.

Methods

To determine reasons for the poor response and better therapeutic approaches we examined the minimal inhibitor concentration (MIC, n = 275) and disc-diffusion zone sizes (n = 205) for ciprofloxacin and nalidixic acid of V. cholerae O1 strains isolated in Bangladesh from 1994 to 2012, and reexamined data from 161patients infected with Vibrio cholerae O1 recruited in four clinical trials who received single- or multiple-dose ciprofloxacin for treatment of cholera and compared their clinical response to the V. cholerae O1 susceptibility.

Results

Although all 275 isolates of V. cholerae O1 remained susceptible to ciprofloxacin using standard MIC and disc-diffusion thresholds, the MIC90 to ciprofloxacin increased from 0.010 in 1994 to 0.475 μgm/ml in 2012. Isolates became frankly resistant to nalidixic with the MIC90 increasing from 21 μgm/ml in 1994 to >256 μgm/ml and 166 of 205 isolates from 1994 to 2005 being frankly resistant using disc-diffusion testing. Isolates resistant to nalidixic acid by disc-diffusion testing had a median ciprofloxacin MIC of 0.190 μgm/ml (10th-90th centiles 0.022 to 0.380); nalidixic acid-susceptible isolates had a median ciprofloxacin MIC of 0.002 (0.002 to 0.012).The rate of clinical success with single-dose ciprofloxacin treatment for nalidixic acid-susceptible strains was 94% (61 of 65 patients) and bacteriologic success 97% (63/65) compared to 18% (12/67) and 8% (5/67) respectively with nalidixic acid-resistant strains (P<0.001 for both comparisons). Multiple-dose treatment with ciprofloxacin had 86% and 100% clinical and bacteriologic success rates respectively in patients infected with nalidixic acid-susceptible strains of V. cholerae O1 compared to clinical success 67% and bacteriologic success 60% with nalidixic acid-resistant strains.

Conclusions

Single-dose ciprofloxacin is not effective for treating cholera caused by V. cholerae O1 with diminished susceptibility to ciprofloxacin, and nalidixic acid disc-diffusion testing effectively screens for such isolates.  相似文献   

13.

Background

In spite of Argentina having one of the highest frequencies of haemolytic uraemic syndrome (HUS), the incidence of Escherichia coli O157:H7 is low in comparison to rates registered in the US. Isolation of several non-O157 shiga toxin-producing Escherichia coli (STEC) strains from cattle and foods suggests that E. coli O157:H7 is an uncommon serotype in Argentina. The present study was undertaken to compare the survival rates of selected non-O157 STEC strains under acidic and alcoholic stress conditions, using an E. coli O157:H7 strain as reference.

Results

Growth at 37°C of E. coli O26:H11, O88:H21, O91:H21, O111:H-, O113:H21, O116:H21, O117:H7, O157:H7, O171:H2 and OX3:H21, was found to occur at pH higher than 4.0. When the strains were challenged to acid tolerance at pH as low as 2.5, viability extended beyond 8 h, but none of the bacteria, except E. coli O91:H21, could survive longer than 24 h, the autochthonous E. coli O91:H21 being the more resistant serotype. No survival was found after 24 h in Luria Bertani broth supplemented with 12% ethanol, but all these serotypes were shown to be very resistant to 6% ethanol. E. coli O91:H21 showed the highest resistance among serotypes tested.

Conclusions

This information is relevant in food industry, which strongly relies on the acid or alcoholic conditions to inactivate pathogens. This study revealed that stress resistance of some STEC serotypes isolated in Argentina is higher than that for E. coli O157:H7.  相似文献   

14.

Background  

Listeria monocytogenesis a well-characterized food-borne pathogen that infects pregnant women and immunocompromised individuals. Listeriolysin O (LLO) is the major virulence factor of the pathogen and is often used as a diagnostic marker for detection of L. monocytogenes. In addition, LLO represents a potent antigen driving T cell-mediated immunity during infection. In the present work, Lactococcus lactisNZ9000 was used as an expression host to hyper-produce LLO under inducible conditions using the NICE (NIsin Controlled Expression) system. We created a modified pNZ8048 vector encoding a six-His-tagged LLO downstream of the strong inducible PnisA promoter.  相似文献   

15.

Background  

Bacillus anthracis is an animal and human pathogen whose virulence is characterized by lethal and edema toxin, as well as a poly-glutamic acid capsule. In addition to these well characterized toxins, B. anthracis secretes several proteases and phospholipases, and a newly described toxin of the cholesterol-dependent cytolysin (CDC) family, Anthrolysin O (ALO).  相似文献   

16.

Aim

In this study, the effects of the addition of salt to treatment with acids (one of several organic acids and salt in various solutions including rich or minimal broth, buffer, or distilled water) on the reduction of Escherichia coli O157:H7 were investigated. The protein expression profiles corresponding to acid stress (acetic acid) with or without salt addition were studied using a comparative proteomic analysis of E. coli O157:H7.

Methods and Results

When acetic, lactic, or propionic acid was combined with 3% NaCl, mutually antagonistic effects of acid and salt on viability of E. coli O157:H7 were observed only in tryptone and yeast extract broth. After exposure to acetic acid alone or in combination with salt, approximately 851 and 916 protein spots were detected, respectively. Analysis of 10 statistically significant differentially expressed proteins revealed that these proteins are mainly related to energy metabolism.

Conclusions

When we compared protein expression of E. coli O157:H7 treated with acetic acid and the combination of the acid and salt, the differentially expressed proteins were not related to acid stress‐ and salt stress‐inducible proteins such as stress shock proteins.

Significance and Impact of the Study

According to these results, the increased resistance of E. coli O157:H7 to acetic acid after the addition of salt may not be the result of synthesis of proteins related to these phenomena; therefore, further research needs to be conducted to identify the mechanism of the mutually antagonistic effect of some organic acids and salt.  相似文献   

17.

Background  

E. coli O157 is a bacterial pathogen that is shed by cattle and can cause severe disease in humans. Phage type (PT) 21/28 is a subtype of E. coli O157 that is found across Scotland and is associated with particularly severe human morbidity.  相似文献   

18.

Background  

Escherichia coli serogroup O157:H7 has emerged as an important zoonotic bacterial pathogen, causing a range of symptoms from self-limiting bloody diarrhea to severe hemorrhagic colitis and hemolytic-uremic syndrome in humans. Beef and dairy cattle are considered the most important animal reservoirs for this pathogen. One of the important virulence characteristics of E. coli O157:H7 is the eaeA gene encoding the 97 kDa surface protein intimin. Intimin is required for attachment and effacement during the interaction of enterohemorrhagic E. coli with human and bovine neonatal enterocytes. The present study was undertaken to test the hypothesis that an adaptive mucosal immune response directed against intimin will reduce or prevent enteric colonization and fecal shedding of E. coli O157:H7 in cattle.  相似文献   

19.

Aims

The present study was carried out to screen the phylloplane bacteria from tea for antagonism against grey blight caused by Pestalotiopsis theae and blister bight caused by Exobasidium vexans and to further evaluate the efficient isolates for disease control potential under field condition.

Methods and Results

A total of 316 morphologically different phylloplane bacteria were isolated. Among the antagonists, the isolates designated as BMO‐075, BMO‐111 and BMO‐147 exhibited maximum inhibitory activity against both the pathogens under in vitro conditions and hence were selected for further evaluation under microplot field trial. Foliar application of 36‐h‐old culture of BMO‐111 (1 × 108 colony‐forming units ml?1) significantly reduced the blister blight disease incidence than the other isolates. The culture of BMO‐111 as well as its culture filtrate effectively inhibited the mycelial growth of various fungal plant pathogens. The isolate BMO‐111 was identified as Ochrobactrum anthropi based on the morphological and 16S rDNA sequence analyses.

Conclusions

It could be concluded that the biocontrol agent O. anthropi BMO‐111 was effective against blister blight disease of tea.

Significance and Impact of the Study

Further study is required to demonstrate the mechanism of its action and formulation for the biocontrol potential against blister blight disease of tea.  相似文献   

20.

Background  

Several cases of human infection caused by verotoxin-producing Escherichia coli (VTEC) O157:H7 in Sweden have been connected with cattle farm visits. Between 1996 and 2002, 18 farms were classified as the source of human cases with isolation of EHEC (Enterohaemorrhagic Escherichia coli) after VTEC O157:H7 had been isolated from cattle on those farms.  相似文献   

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