A Microscopic Method to Visualize Escherichia coli Interaction with Beef Muscle

Prachaiyo, P.; McLandsborough, Lynne

doi:10.4315/0362-028x-63.4.427

Cited by 30 publications

(19 citation statements)

References 21 publications

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“…It has been shown before that bacterial strains with similar hydrophobic properties show similar adhesion patterns to surfaces and that hydrophobic surfaces favor adhesion of hydrophobic bacteria (21). The relative hydrophobicity of the bacterial strains used in this work was measured using their adhesion to both hexadecane and xylene, since the results of hydrocarbon attachment measurements may vary depending on the hydrocarbons tested (20). In this study, the attachment results were similar for hexadecane and xylene, as shown by the data in Table 1.…”

Section: Resultssupporting

confidence: 62%

“…These values represent the percentage of cells that did not adhere to the hydrocarbons and remained in the aqueous phase. Cells are considered relatively hydrophobic when the values of the OD 540 ratios are closer to 0 (indicating fewer cells remaining in aqueous phase) and relatively hydrophilic when the values are closer to 1 (20). Experiments were conducted in triplicate, and the values reported are average values with the standard deviation of three measurements.…”

Section: Methodsmentioning

confidence: 99%

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Effect of Micro- and Nanoscale Topography on the Adhesion of Bacterial Cells to Solid Surfaces

Hsu

Fang

Borca-Tasciuc

et al. 2013

Appl Environ Microbiol

292

212

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c Attachment and biofilm formation by bacterial pathogens on surfaces in natural, industrial, and hospital settings lead to infections and illnesses and even death. Minimizing bacterial attachment to surfaces using controlled topography could reduce the spreading of pathogens and, thus, the incidence of illnesses and subsequent human and financial losses. In this context, the attachment of key microorganisms, including Escherichia coli, Listeria innocua, and Pseudomonas fluorescens, to silica and alumina surfaces with micron and nanoscale topography was investigated. The results suggest that orientation of the attached cells occurs preferentially such as to maximize their contact area with the surface. Moreover, the bacterial cells exhibited different morphologies, including different number and size of cellular appendages, depending on the topographical details of the surface to which they attached. This suggests that bacteria may utilize different mechanisms of attachment in response to surface topography. These results are important for the design of novel microbe-repellant materials.

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Section: Resultssupporting

confidence: 62%

Section: Methodsmentioning

confidence: 99%

Effect of Micro- and Nanoscale Topography on the Adhesion of Bacterial Cells to Solid Surfaces

Hsu

Fang

Borca-Tasciuc

et al. 2013

Appl Environ Microbiol

292

212

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“…GFP-expressing bacteria have been previously used in a number of studies that did not involve intact, living animals (21)(22)(23)(24)(25)(26)(27)(28)(29)). An example of such studies was the visualization of the in vitro infection of muscle tissue by the pathogenic E. coli O157H GFP (28).…”

Section: Discussionmentioning

confidence: 99%

“…An example of such studies was the visualization of the in vitro infection of muscle tissue by the pathogenic E. coli O157H GFP (28). Another approach examined the mouse gastrointestinal tract after gavage infection by removal and fixation of the gastrointestinal tissue (29).…”

Section: Discussionmentioning

confidence: 99%

Spatial–temporal imaging of bacterial infection and antibiotic response in intact animals

Zhao

Yang

Baranov

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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We describe imaging the luminance of green fluorescent protein (GFP)-expressing bacteria from outside intact infected animals. This simple, nonintrusive technique can show in great detail the spatial-temporal behavior of the infectious process. The bacteria, expressing the GFP, are sufficiently bright as to be clearly visible from outside the infected animal and recorded with simple equipment. Introduced bacteria were observed in several mouse organs including the peritoneal cavity, stomach, small intestine, and colon. Instantaneous real-time images of the infectious process were acquired by using a color charge-coupled device video camera by simply illuminating mice at 490 nm. Most techniques for imaging the interior of intact animals may require the administration of exogenous substrates, anesthesia, or contrasting substances and require very long data collection times. In contrast, the whole-body fluorescence imaging described here is fast and requires no extraneous agents. The progress of Escherichia coli-GFP through the mouse gastrointestinal tract after gavage was followed in realtime by whole-body imaging. Bacteria, seen first in the stomach, migrated into the small intestine and subsequently into the colon, an observation confirmed by intravital direct imaging. An i.p. infection was established by i.p. injection of E. coli-GFP. The development of infection over 6 h and its regression after kanamycin treatment were visualized by whole-body imaging. This imaging technology affords a powerful approach to visualizing the infection process, determining the tissue specificity of infection, and the spatial migration of the infectious agents.green fluorescent protein ͉ external optical imaging ͉ Escherichia coli ͉ antibiotic response ͉ mice M any biological research techniques depend on detecting uniquely distinguishable cells, e.g., cells exhibiting unique or aberrant morphologies. Far greater sensitivity is achieved by adding exogenous markers, especially those that are optically visible. The techniques of early embryologists marking developmental pathways with India ink injections have evolved into modern methods that include genetically transducing cells with reporters such as dye-activating enzymes. However, such methods still could not detect small, rare targets and usually could not be visualized in living systems. A qualitative advance in sensitivity is afforded by the new class of cell marking reagents exemplified by green fluorescent protein (GFP) and its derivatives. Genetically based, these can be permanent and nontoxic, and are very strongly fluorescent. Most importantly, because their presence seems to have little effect on the marked cell, observations in living tissue are now possible. We show here that GFP enables previously impossible, noninvasive tracking of bodily disease agents in space and time.GFP has been used as a reporter gene for numerous biological processes (1). Its freedom from required substrates or cofactors allows it to be expressed and visualized in living cells with no apparent...

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“…Salmonella/pEGFP strains were only grown on media supplemented with antibiotic. Prachaiyo and McLandsborough (27) found that when grown without antibiotic selection, approximately 90% of the cell population lost the plasmid over the course of an 8-h culture. Transformed Salmonella strains inoculated onto LBA-amp plates, incubated for 12 to 16 h, and stored at 4ЊC stably maintained the pEGFP plasmid for over 30 days.…”

Section: Stability Of Egfp Expression By Salmonella Strainsmentioning

confidence: 99%

Survival of Salmonella Transformed To Express Green Fluorescent Protein on Italian Parsley as Affected by Processing and Storage

Duffy

Cisneros‐Zevallos

Castillo

et al. 2005

Journal of Food Protection

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To study the effect of processing and storage parameters on the survival of Salmonella on fresh Italian parsley, parsley bunches were dipped for 3 or 15 min in suspensions that were preequilibrated to 5, 25, or 35ЊC and inoculated with Salmonella transformed to express enhanced green fluorescent protein. Loosely attached and/or associated, strongly attached and/or associated, and internalized and/or entrapped Salmonella cells were enumerated over 0, 1, and 7 days of storage at 25ЊC and over 0, 1, 7, 14, and 30 days of storage at 4ЊC using surface-plating procedures. Leaf sections obtained from samples after 0, 1, and 7 days of storage were examined using confocal scanning laser microscopy. Temperature of the dip suspension had little effect on the attachment and survival of Salmonella cells on parsley. Regardless of the temperature or duration of dip, Salmonella was internalized. Immersion for longer times resulted in higher numbers of attached and internalized cells. Microscopic observations supported these results and revealed Salmonella cells near the stomata and within cracks in the cuticle. Storage temperature had the greatest impact on the survival of Salmonella cells on parsley. When stored at 25ЊC, parsley had a shelf life of 7 days, and Salmonella populations significantly increased over the 7 days of storage. For parsley stored at 4ЊC, numbers of Salmonella cells decreased over days 0, 1, and 7. After 7 days of storage, there were no viable internalized Salmonella cells detected. Storage temperature represents an important control point for the safety of fresh parsley.Fresh produce is an important source of enteric pathogens in foodborne illness outbreaks. Fruits and vegetables, which are commonly consumed without any cooking treatment, may potentially harbor many foodborne pathogens (12,21). Ingestion of fresh parsley has been associated with Shigella sonnei and enterotoxigenic Escherichia coli infections (23). Salmonellosis outbreaks have been epidemiologically associated with the consumption of cantaloupes, watermelon, tomatoes, lettuce, celery, alfalfa sprouts, parsley, unpasteurized orange juice, and other raw salad vegetables (10,11,15,17,20). To assess the risk factors involved in the contamination of fresh produce with human pathogens, it is important to understand the microbial ecology of the various products and possible vectors during harvesting and processing that may contribute to product contamination.Cultivation, harvesting, and retail preparation of fruits and vegetables provide numerous opportunities for human and equipment contact (5, 18). Typical manual labor operations including picking, sorting, trimming, tying, bunching, and removal of visible soil in rinse water, all of which subject the product to extensive handling and exposure to a variety of microorganisms. Rinse water can be of questionable bacteriological quality (18). Reina et al. (28) found that recycled water developed Enterobacteriaceae populations of up to 10 6 CFU/g during a typical day's operation. Human pat...

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A Microscopic Method to Visualize Escherichia coli Interaction with Beef Muscle

Cited by 30 publications

References 21 publications

Effect of Micro- and Nanoscale Topography on the Adhesion of Bacterial Cells to Solid Surfaces

Effect of Micro- and Nanoscale Topography on the Adhesion of Bacterial Cells to Solid Surfaces

Spatial–temporal imaging of bacterial infection and antibiotic response in intact animals

Survival of Salmonella Transformed To Express Green Fluorescent Protein on Italian Parsley as Affected by Processing and Storage

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