The Detection of Microorganisms and Organic Material on Stainless Steel Food Contact Surfaces

Few methods are available for the differentiation of microorganism and organic material on surfaces, although such mixtures are commonplace, particularly in the food industry, where food debris (soil) and microorganisms frequently foul food contact surfaces and pose challenges in terms of hygiene and cleanability. It would be of value to discern any differences in removal or persistence on surfaces. This review considers some methods which are available. Direct epifluorescence microscopy (DEM) enables visual differentiation, but traditional microbiological culture methods cannot detect organic soil. Atomic force microscopy (AFM) provides images of the surface on the nanometer scale, with minimal preparation, and is able to visualise both cellular and acellular components of the mixture, particularly prior to cleaning. Confocal laser scanning microscopy (CLSM) also has potential in this area. Surface sensitive methods such as X-Ray photoelectron spectroscopy (XPS) and Time-of-flight secondary ion mass spectrometry (ToFSIMS) provide information on chemical species present on a surface. Those chemical species more likely on microbial cells may be differentiated from those more likely in a specified organic soil, thus comparisons may be made as to differential removal of the organic soil and microbial cells. These methods may be of value in studies on the fouling and cleanability of surfaces.     

Comparison of fluorinated polymers against stainless steel, glass and polypropylene in microbial biofilm adherence and removal

Biofilm formation is a long-standing problem in ultrapure water and bioprocess fluid transport lines. The standard materials used in these applications (316L stainless steel, polypropylene and glass) have long been known to be good surfaces for the attachment of bacteria and other biological materials. To compare the relative tenacity of biofilms grown on materials used in manufacturing processes, a model system for biofilm attachment was constructed that approximates the conditions in industrial process systems. New fluorinated polymers were compared to the above materials by evaluating the surface area coverage of bacterial populations on materials before and after mild chemical treatment. In addition, contact angle studies compared the relative hydrophobicity of surfaces to suspensions of bacteria in growth media, and scanning electron microscopy and atomic force microscopy studies were used to characterize surface smoothness and surface defects. Biofilm adherence to polymer-based substrata was determined to be a function of both surface finish and surface chemistry. Specifically, materials that are less chemically reactive, as indicated by higher contact angle, can have rougher surface finishes and still be amenable to biofilm removal. Received 20 March 1997/ Accepted in revised form 15 July 1997     

Surface-active antifungal polyquaternary amine

There is a distinct need for antimicrobial compounds that can act at surfaces without leaching into the environment. Such materials should be easy to synthesize, be easy to apply to surfaces, and display reasonable levels of antimicrobial and antifungal activity. Here we describe such a surface-active compound and demonstrate its ability to inhibit the growth of the filamentous fungus Aspergillus niger. We have synthesized a series of polyquaternary ammonium compounds by atom transfer radical polymerization. Two members of this series were tested for their ability to inhibit the growth of A. niger. The compounds were dried onto surfaces, and the treated surfaces were then used as growth chambers for A. niger. A water soluble polyquaternary amine compound was shown to effectively kill A. niger in solution in a dose-dependent manner. Conversely, a water insoluble polyquaternary amine compound was shown to kill only the fungi in direct contact with the material on the surface. These results have important implications for the development of effective, environmentally benign, surface-active anti-fungal compounds.     

Hepatitis A virus attachment to agri-food surfaces using immunological, virological and thermodynamic assays

AIMS: This study was designed to investigate the ability of hepatitis A virus (HAV) to attach to various food contact surfaces. METHODS AND RESULTS: HAV attachment was demonstrated after elution of attached viruses from solid surfaces by an immunofluorescent method using anti-HAV-specific antibodies and confocal microscopy. Attachment and survival of HAV on stainless steel, copper, polythene and polyvinyl chloride (PVC) at 20 and 4 degrees C after 2 and 4 h were quantified by plaque assay. HAV was shown to attach almost instantaneously to all four surfaces tested. Attachment of HAV depended on initial viral concentration and was slightly greater at 4 degrees C. The total surface energy (gammaTOT), nonpolar Lifshitz-Van der Waals (gammaLW) and polar short range (gammaSR) hydrogen-bonding components for HAV and each surface as well as total free energy of the system were determined by contact angle measurements using an extended Young equation [Young (1805) Philosophical Transactions of The Royal Society (London) 95, 65-87). The calculation of these parameters predicted the favourable conditions for attachment of HAV to all four surfaces tested. CONCLUSION: HAV particles attach to stainless steel, copper, polythene and PVC at 20 and 4 degrees C and the total free energy of the interaction is optimal for this attachment. SIGNIFICANCE AND IMPACT OF THE STUDY: Comprehension of viral attachment to the solid surfaces will permit to successfully disinfect these surfaces and to establish a better surveillance programme for control of viral food-borne illnesses.     

Cleanability in relation to bacterial retention on unused and abraded domestic sink materials

The relative cleanability of stainless steel, enamelled steel, mineral resin and polycarbonate domestic sink materials was assessed by comparing the number of organisms remaining on surfaces after cleaning. In unused condition all materials, other than one enamelled steel, were equally cleanable. Stainless steel, abraded artificially or impact damaged to a similar degree as stainless steel subjected to domestic wear, retained approximately one log order less bacteria after cleaning than the other materials subjected to the same treatments. Little difference in cleanability was recorded between the abraded surfaces of the other materials although enamelled steel surfaces were less cleanable than mineral resin or polycarbonate after impact damage, because of the greater susceptibility of enamelled steel to damage by this treatment. When cleaning time was extended beyond 10 s for the abraded and impact damaged materials, their cleanability was not enhanced as compared with stainless steel. Changes in surface finish after abrasion were assessed by surface roughness measurement and scanning electron microscopy. Surfaces with poor cleanability before and after abrasion were characterized by pitting, crevices or jags. These surfaces are likely to retain more bacteria because of increased numbers of attachment sites, a larger bacterial/material surface contact area and topographical areas in which applied cleaning shear forces are reduced. Materials that resist surface changes, e.g. stainless steel, will remain more hygienic when subjected to natural wear than materials which become more readily damaged.     

Exposure of Escherichia coli ATCC 12806 to Sublethal Concentrations of Food-Grade Biocides Influences Its Ability To Form Biofilm,Resistance to Antimicrobials,and Ultrastructure

Escherichia coli ATCC 12806 was exposed to increasing subinhibitory concentrations of three biocides widely used in food industry facilities: trisodium phosphate (TSP), sodium nitrite (SNI), and sodium hypochlorite (SHY). The cultures exhibited an acquired tolerance to biocides (especially to SNI and SHY) after exposure to such compounds. E. coli produced biofilms (as observed by confocal laser scanning microscopy) on polystyrene microtiter plates. Previous adaptation to SNI or SHY enhanced the formation of biofilms (with an increase in biovolume and surface coverage) both in the absence and in the presence (MIC/2) of such compounds. TSP reduced the ability of E. coli to produce biofilms. The concentration of suspended cells in the culture broth in contact with the polystyrene surfaces did not influence the biofilm structure. The increase in cell surface hydrophobicity (assessed by a test of microbial adhesion to solvents) after contact with SNI or SHY appeared to be associated with a strong capacity to form biofilms. Cultures exposed to biocides displayed a stable reduced susceptibility to a range of antibiotics (mainly aminoglycosides, cephalosporins, and quinolones) compared with cultures that were not exposed. SNI caused the greatest increase in resistances (14 antibiotics [48.3% of the total tested]) compared with TSP (1 antibiotic [3.4%]) and SHY (3 antibiotics [10.3%]). Adaptation to SHY involved changes in cell morphology (as observed by scanning electron microscopy) and ultrastructure (as observed by transmission electron microscopy) which allowed this bacterium to persist in the presence of severe SHY challenges. The findings of the present study suggest that the use of biocides at subinhibitory concentrations could represent a public health risk.     

Antibacterial isoeugenol coating on stainless steel and polyethylene surfaces prevents biofilm growth

Aims

Pathogenic bacteria can spread between individuals or between food items via the surfaces they share. Limiting the survival of pathogens on surfaces, therefore, presents an opportunity to limit at least one route of how pathogens spread. In this study, we propose that a simple coating with the essential oil isoeugenol can be used to circumvent the problem of bacterial transfer via surfaces.

Methods and Results

Two commonly used materials, stainless steel and polyethylene, were coated by physical adsorption, and the coatings were characterized by Raman spectroscopy, atomic force microscopy and water contact angle measurements. We quantified and visualized the colonization of coated and uncoated surfaces by three bacteria: Staphylococcus aureus, Listeria monocytogenes and Pseudomonas fluorescens. No viable cells were detected on surfaces coated with isoeugenol.

Conclusions

The isoeugenol coating prepared with simple adsorption proved effective in preventing biofilm formation on stainless steel and polyethylene surfaces. The result was caused by the antibacterial effect of isoeugenol, as the coating did not diminish the adhesive properties of the surface.

Significance and Impact of the Study

Our study demonstrates that a simple isoeugenol coating can prevent biofilm formation of S. aureus, L. monocytogenes and P. fluorescens on two commonly used surfaces.     

Removal and transfer of viruses on food contact surfaces by cleaning cloths

Contamination of food contact surfaces with pathogens is considered an important vehicle for the indirect transmission of food-borne diseases. Five different cleaning cloths were assessed for the ability to remove viruses from food contact surfaces (stainless steel surface and nonporous solid surface) and to transfer viruses back to these surfaces. Cleaning cloths evaluated include two different cellulose/cotton cloths, one microfiber cloth, one nonwoven cloth, and one cotton terry bar towel. Four viral surrogates (murine norovirus [MNV], feline calicivirus [FCV], bacteriophages PRD1 and MS2) were included. Removal of FCV from stainless steel was significantly greater (P ≤ 0.05) than that from nonporous solid surface, and overall removal of MNV from both surfaces was significantly less (P ≤ 0.05) than that of FCV and PRD1. Additionally, the terry towel removed significantly fewer total viruses (P ≤ 0.05) than the microfiber and one of the cotton/cellulose cloths. The cleaning cloth experiments were repeated with human norovirus. For transfer of viruses from cloth to surface, both cellulose/cotton cloths and microfiber transferred an average of 3.4 and 8.5 total PFU, respectively, to both surfaces, and the amounts transferred were significantly different (P ≤ 0.05) from those for the nonwoven cloth and terry towel (309 and 331 total PFU, respectively). There was no statistically significant difference (P > 0.05) in the amount of virus transfer between surfaces. These data indicate that while the cleaning cloths assessed here can remove viruses from surfaces, some cloths may also transfer a significant amount of viruses back to food contact surfaces.     

Cleanability in relation to bacterial retention on unused and abraded domestic sink materials

H olah , J.T. & T horpe , R.H. 1990. Cleanability in relation to bacterial retention on unused and abraded domestic sink materials. Journal of Applied Bacteriology 69 , 599–608.
The relative Cleanability of stainless steel, enamelled steel, mineral resin and polycarbonate domestic sink materials was assessed by comparing the number of organisms remaining on surfaces after cleaning. In unused condition all materials, other than one enamelled steel, were equally cleanable. Stainless steel, abraded artificially or impact damaged to a similar degree as stainless steel subjected to domestic wear, retained approximately one log order less bacteria after cleaning than the other materials subjected to the same treatments. Little difference in Cleanability was recorded between the abraded surfaces of the other materials although enamelled steel surfaces were less cleanable than mineral resin or polycarbonate after impact damage, because of the greater susceptibility of enamelled steel to damage by this treatment. When cleaning time was extended beyond 10 s for the abraded and impact damaged materials, their Cleanability was not enhanced as compared with stainless steel. Changes in surface finish after abrasion were assessed by surface roughness measurement and scanning electron microscopy. Surfaces with poor Cleanability before and after abrasion were characterized by pitting, crevices or jags. These surfaces are likely to retain more bacteria because of increased numbers of attachment sites, a larger bacterial/material surface contact area and topographical areas in which applied cleaning shear forces are reduced. Materials that resist surface changes, e.g. stainless steel, will remain more hygienic when subjected to natural wear than materials which become more readily damaged.     

Increased endothelial cell adhesion on plasma modified nanostructured polymeric and metallic surfaces for vascular stent applications

Techniques to regenerate the vasculature have risen considerably over the last few decades due to the increased clinical diagnosis of artery narrowing and blood vessel blockage. Although initially re‐establishing blood flow, current small diameter vascular regenerative materials often eventually cause thrombosis and restenosis due to a lack of initial endothelial cell coverage on such materials. The objective of this in vitro study was to evaluate commonly used vascular materials (specifically, polyethylene terephthalate, polytetrafluoroethylene, polyvinyl chloride, polyurethane, nylon, commercially pure titanium, and a titanium alloy (Ti6Al4V)) modified using an ionic plasma deposition (IPD) process and a nitrogen ion implantation plasma deposition (NIIPD) process. Such surface modifications have been previously shown to create nanostructured surface features which mimic the natural nanostructured surface features of blood vessels. The modified and unmodified surfaces were characterized by scanning electron microscopy, atomic force microscopy and surface energy measurements. Furthermore, in vitro endothelial cell adhesion tests (a key first step for vascular material endothelialization) demonstrated increased endothelial cell adhesion on many modified (with IPD and NIIPD + IPD) compared to unmodified samples. In general, endothelial cell adhesion increased with nanoroughness and surface energy but demonstrated a decreased endothelial cell adhesion trend after an optimal coating surface energy value was reached. Thus, results from this study provided materials and a versatile surface modification process that can potentially increase endothelialization faster than current unmodified (conventional) polymer and metallic vascular materials. Biotechnol. Bioeng. 2009;103: 459–471. © 2009 Wiley Periodicals, Inc.     

Attachment forces of pea aphids (Acyrthosiphon pisum) on different legume species

1. Sympatric populations of insects adapted to different host plants are good model systems not only to study how they adapt to the chemistry of their food plant, but also to investigate whether morphological modifications evolved enabling them to live successfully on a certain plant species. 2. The pea aphid, Acyrthosiphon pisum (Harris) encompasses at least 11 genetically distinct sympatric host races, each showing a preference for a certain legume species. The leaflet surfaces of these legumes differ considerably in their wax coverage. 3. It was investigated whether the attachment structures of three pea aphid genotypes from different host races are adapted to the different surface properties of their host plants and whether they show differences in their attachment ability on the respective host and non‐host plants. 4. The surface morphology of plants and aphid tarsi was examined using SEM (scanning electron microscopy). The ability of the aphids to walk on specific surfaces was tested using traction force measurements. 5. The presence of wax blooms on the leaflets lowers the aphids' attachment ability considerably and diminishes their subsequent attachment on ‘neutral’ surfaces like glass. The pea aphid host races differ in their ability to walk on certain surfaces. However, the genotype from the adapted aphid host race was not necessarily the one with the best walking performance on their host plant. All aphids, regardless of the original host plant, were most efficient on the neutral control surface glass. The general host plant Vicia faba was the plant with the most favourable surface for all aphid host races.     

Towards bioinspired superhydrophobic poly(L-lactic acid) surfaces using phase inversion-based methods

The water repellency and self-cleaning ability of many biological surfaces has inspired many fundamental and practical studies related to the development of synthetic superhydrophobic surfaces. However, the investigation of such substrates made of biodegradable polymers has been scarce. Simple approaches based on a single step, performed at room temperature (and pressure), were implemented to obtain superhydrophobic poly(L-lactic acid) (PLLA) surfaces via phase inversion-based methods, without addition of low-surface-energy compounds. Water contact angles above 150 degrees were obtained using some processing conditions. In such cases scanning electronic microscopy micrographs of such surfaces revealed a clear rough texture composed by leafy clusters with micro-nano binary structures. Such materials could be used in specific environmental and biomedical applications, namely in implantable materials or in antibacterial or antithrombogenic surfaces.     

Effect of Ionic Strength on Initial Interactions of Escherichia coli with Surfaces, Studied On-Line by a Novel Quartz Crystal Microbalance Technique

A novel quartz crystal microbalance (QCM) technique was used to study the adhesion of nonfimbriated and fimbriated Escherichia coli mutant strains to hydrophilic and hydrophobic surfaces at different ionic strengths. This technique enabled us to measure both frequency shifts (Deltaf), i.e., the increase in mass on the surface, and dissipation shifts (DeltaD), i.e., the viscoelastic energy losses on the surface. Changes in the parameters measured by the extended QCM technique reflect the dynamic character of the adhesion process. We were able to show clear differences in the viscoelastic behavior of fimbriated and nonfimbriated cells attached to surfaces. The interactions between bacterial cells and quartz crystal surfaces at various ionic strengths followed different trends, depending on the cell surface structures in direct contact with the surface. While Deltaf and DeltaD per attached cell increased for nonfimbriated cells with increasing ionic strengths (particularly on hydrophobic surfaces), the adhesion of the fimbriated strain caused only low-level frequency and dissipation shifts on both kinds of surfaces at all ionic strengths tested. We propose that nonfimbriated cells may get better contact with increasing ionic strengths due to an increased area of contact between the cell and the surface, whereas fimbriated cells seem to have a flexible contact with the surface at all ionic strengths tested. The area of contact between fimbriated cells and the surface does not increase with increasing ionic strengths, but on hydrophobic surfaces each contact point seems to contribute relatively more to the total energy loss. Independent of ionic strength, attached cells undergo time-dependent interactions with the surface leading to increased contact area and viscoelastic losses per cell, which may be due to the establishment of a more intimate contact between the cell and the surface. Hence, the extended QCM technique provides new qualitative information about the direct contact of bacterial cells to surfaces and the adhesion mechanisms involved.     

Effect of nanostructure on wettability on copper surface: a molecular dynamic study

Molecular dynamics simulations (MDS) are employed to investigate the effects of interatomic interaction and nanostructure on wettability of water on a copper plate. In the nano scale, these simulation results showed that the contact angle gradually increases with the decreasing of the reaction parameters, which results in the decreasing of free energy on the solid-liquid interface. Therefore, it leads to that the hydrophilic material is turned into hydrophobic, which fits the results that the wettability is changed by low surface energy materials in macro scale. Furthermore, the contact angles on smooth and rough surfaces are 87° and 71.6°, respectively. That is to say that the hydrophilic will increase for hydrophilic material due to the existence of one-layer structure; it agrees with the experimental results in macro scale.     

鼠伤寒沙门菌pStSR100质粒对生物膜形成的影响

目的:鼠伤寒沙门菌在多种表面形成的生物膜对其致病性和引起食物中毒等方面起着重要作用,本研究探讨鼠伤寒沙门菌pStSR100质粒对细菌在不同材质表面生物膜形成的影响。方法:用LB(Luria-Bertani,LB)培养基和TSB(Tryptose Soya Broth,TSB)培养基分别将携带pStSR100质粒的野生株在96孔板与放置无菌小圆玻片的24孔板中静态培养48 h,用结晶紫半定量法确定生物膜形成的适宜培养基。将野生株与消除质粒的突变株,用结晶紫半定量法和激光共聚焦显微镜(Confocal Laser scanning microscopy,CLSM)观察其在聚苯乙烯培养板和小圆玻片表面形成生物膜的差异。结果:用LB培养时细菌生物膜的形成能力高于用TSB培养,LB培养基更适宜生物膜形成;结晶紫半定量法结果表明野生株比突变株在小圆玻片表面形成生物膜的能力明显增强,而在聚苯乙烯培养板表面两者则无明显差异;CLSM观察发现,野生株在小圆玻片表面形成融合成片的大克隆,突变株仅形成较小克隆。结论:鼠伤寒沙门菌pStSR100质粒能促进该菌在亲水性材质表面生物膜的形成,但其对该菌在疏水性材质表面生物膜的形成未见明显影响,这一新发现为进一步研究鼠伤寒沙门菌生物膜形成的调控机制,研制抗感染材料提供了理论和实验依据。     

Microscopic investigations of the interaction of proteins with surfaces

The application of light microscopy to the study of protein-surface interactions is described with several examples showing its versatility. Extensive use is made of image analysis algorithms to extract quantitative information from the digital images and it is shown how this can shed light on the processes taking place on the surface. Amongst the surfaces investigated are siloxane films on silicon wafers, glucose oxidase/poly(ethyleneglycol) films cast onto glass slides and glucose oxidase entrapped in electropolymerised poly(phenyleneoxide) films on platinum. Parameters that can be measured include: surface loading, surface heterogeneity, distribution of enzyme activity, surface mobility and film porosity. Both reflected light and fluorescence microscopy were used to characterise the surfaces and it was concluded that not only does light microscopy offer a powerful tool for the characterisation of surfaces but it may also provide an interface between bioelectronic materials and conventional computing machinery.     

Leaching of plasticizers from and surface characterization of PVC blood platelet bags.

The leaching of phthalate plasticizers from four types of blood platelet bags was investigated. The anticoagulant solutions used in the blood collection bags had pH values of 5.64 +/- .04 and contained no detectable amounts of phthalates. Platelet bag materials from each bag were soaked in normal salines for up to 5 days. The salines were tested for the leached phthalates from the bags but none could be found. However, di-(2-ethylhexyl) phthalate (DEHP) leached out of the PL-146 and Terumo bags into bovine calf serum used for soaking the bag materials. There was an increase in the amount of DEHP leached from about 1.1 mg at the end of one day to about 3.3 mg per gm of bag material at the end of a five day extraction with the serum. In PL-732 sets, a platelet bag made of a specialty polyolefin, the amount of DEHP leached out was less than 0.02 mg per g of bag material. CLX bags, which contained tri-(2-ethylhexyl) trimelliate (TETM) as a plasticizer, showed a negligible amount of it leaching into the calf serum. Infra-red spectra showed that PL-146 bags had been coated with a layer of a fatty acid amide while the Terumo bags contained a layer of a silicone fluid on their inner surfaces. CLX bags showed a coating of stearates, which were probably soaps of calcium or zinc. Scanning electron microscopy (SEM) showed that the inner surfaces of each brand of the bag were distinctly different morphologically. The two PVC bags were very similar whereas the surfaces morphology of PL-732 was rougher. Terumo bag had a different surface morphology than those of the other bags whereas the CLX bags had a very regular surface pattern. The exact significance of the surface morphology is not certain but excessively rough surfaces may not be desirable for the bags.     

Response surface optimization for joint contact model evaluation

When optimization is used to evaluate a joint contact model's ability to reproduce experimental measurements, the high computational cost of repeated contact analysis can be a limiting factor. This paper presents a computationally-efficient response surface optimization methodology to address this limitation. Quadratic response surfaces were fit to contact quantities (contact force, maximum pressure, average pressure, and contact area) predicted by a discrete element contact model of the tibiofemoral joint for various combinations of material modulus and relative bone pose (i.e., position and orientation). The response surfaces were then used as surrogates for costly contact analyses in optimizations that minimized differences between measured and predicted contact quantities. The methodology was evaluated theoretically using six sets of synthetic (i.e., computer-generated) contact data, and practically using one set of experimental contact data. For the synthetic cases, the response surface optimizations recovered all contact quantities to within 3.4% error. For the experimental case, they matched all contact quantities to within 6.3% error except for maximum contact pressure, which was in error by up to 50%. Response surface optimization provides rapid evaluation of joint contact models within a limited range of relative bone poses and can help identify potential weaknesses in contact model formulation and/or experimental data quality.     

How do plant waxes cause flies to slide? Experimental tests of wax-based trapping mechanisms in three pitfall carnivorous plants

The waxy surfaces of three carnivorous plants, Nepenthes ventrata (Nepenthaceae), Brocchinia reducta and Catopsis berteroniana (Bromeliaceae), were compared using scanning electron microscopy (SEM). Their effects on attachment and locomotion of the fly Calliphora vomitoria were studied. The waxy surface of N. ventrata is comprised of a heterogeneous layer from which only platelet-shaped crystalloids could be detached by brushing. In the two bromeliads, the crystalloids are thread-shaped and form a homogenous dense network, which was entirely removable from the epidermis. Experimental data showed that none of the flies was able to walk across any of the waxy surfaces and only a few were able to take off from those surfaces. Both the absence of sites for claw anchorage, especially in N. ventrata, and the wax itself were shown to contribute to the trapping ability of the plants. Only half of the flies quickly recovered their locomotion ability on a glass surface after 20 min of being tested on waxy plant surfaces. SEM observations revealed that the wax of C. berteroniana formed a powder of broken crystals on the tenent setae of the flies' pulvilli. In contrast, the waxes of B. reducta and N. ventrata appeared to have lost their crystal structure in contact with the tenent setae and formed an amorphous substance that adhered setae together. We hypothesize that wax interacts with adhesive fluids secreted by the fly pad and thereby prevents the tenent setae from functioning effectively.