Residual Viral and Bacterial Contamination of Surfaces after Cleaning and Disinfection

Environmental surfaces contaminated with pathogens can be sources of indirect transmission, and cleaning and disinfection are common interventions focused on reducing contamination levels. We determined the efficacy of cleaning and disinfection procedures for reducing contamination by noroviruses, rotavirus, poliovirus, parechovirus, adenovirus, influenza virus, Staphylococcus aureus, and Salmonella enterica from artificially contaminated stainless steel surfaces. After a single wipe with water, liquid soap, or 250-ppm free chlorine solution, the numbers of infective viruses and bacteria were reduced by 1 log₁₀ for poliovirus and close to 4 log₁₀ for influenza virus. There was no significant difference in residual contamination levels after wiping with water, liquid soap, or 250-ppm chlorine solution. When a single wipe with liquid soap was followed by a second wipe using 250- or 1,000-ppm chlorine, an extra 1- to 3-log₁₀ reduction was achieved, and except for rotavirus and norovirus genogroup I, no significant additional effect of 1,000 ppm compared to 250 ppm was found. A reduced correlation between reduction in PCR units (PCRU) and reduction in infectious particles suggests that at least part of the reduction achieved in the second step is due to inactivation instead of removal alone. We used data on infectious doses and transfer efficiencies to estimate a target level to which the residual contamination should be reduced and found that a single wipe with liquid soap followed by a wipe with 250-ppm free chlorine solution was sufficient to reduce the residual contamination to below the target level for most of the pathogens tested.     

Cleaning of inorganic membranes after whey and milk ultrafiltration

Cleaning of an inorganic ultrafiltration membrane has been quantified through hydraulic, physicochemical, and spectroscopic (infrared and x-photoelectron spectroscopy) analyses. An efficient cleaning sequence of nitric acid followed by sodium hypochlorite has been proposed for cleaning of defatted whey protein concentrate and milk ultrafiltration membranes. The influence of reversed sequence and time reduction are discussed together with the action of both cleaning chemicals. In spite of residual fouling left after every cleaning sequence studied, hydraulic cleanliness of the membrane was achieved, particularly after the standard procedure.     

Effect of Manual Brush Cleaning on Biomass and Community Structure of Microfouling Film Formed on Aluminum and Titanium Surfaces Exposed to Rapidly Flowing Seawater

Metals exposed to rapidly flowing seawater are fouled by microbes that increase heat transfer resistance. In this study, results of biochemical test methods quantitatively relating the biomass and community structure of the microfouling film on aluminum and titanium to heat transfer resistance across the metal surface during three cycles of free fouling and manual brushing showed that cleaning accelerates the rate of fouling measured as the loss of heat transfer efficiency and as microfouling film biomass. The results also showed that the rate of fouling, measured as an increase in heat transfer resistance, is faster on titanium than on aluminum but that the titanium surface is more readily cleaned. In three cycles of free fouling and cleaning with a stiff-bristle nylon brush, the free-fouling communities re-forming on aluminum became enriched in bacteria containing short-branched fatty acids as the cycling progressed. The free-fouling community on titanium revealed an increasingly diverse morphology under scanning electron microscopy that was enriched in a portion of the microeucaryotes. Brushing removed most of the biomass, but left a residual community that was relatively enriched in a portion of the bacterial assembly containing cyclopropane fatty acids on aluminum and in a more diverse community on the titanium surface. The residual communities left after cleaning of titanium revealed an increase in bacteria with short-branched fatty acids and in microeucaryotes as cleaning continued. No significant changes occurred in the residual microbial community structure left on aluminum with cleaning; it was, again, less diverse than that remaining on titanium. The residual communities secreted a twofold-larger amount of extracellular polymer, measured as the ratio of total organic carbon to lipid phosphate, than did the free-fouling community on both surfaces.     

Effectiveness of cleaning techniques used in the food industry in terms of the removal of bacterial biofilms

The effectiveness of cleaning was investigated through food factory trials and laboratory experiments using a naturally occurring biofilm from a food factory environment and generated biofilms. The efficacy of factory cleaning and disinfection programmes was assessed by swabbing and total viable count (TVC) analysis of surfaces before cleaning, after cleaning and after disinfection. Cleaning produced a 0.91 log reduction in the attached population. Investigation of the effectiveness of a variety of cleaning methods in the removal of a naturally occurring food factory biofilm showed that the high pressure spray and the mechanical floor scrubber, which use a high degree of mechanical action, were most effective. Cleaning trials with biofilms of Pseudomonas aeruginosa or Staphylococcus aureus showed that spraying with water at pressures of 34.5, 51.7 and 68.9 bar did not significantly increase the removal, as assessed by direct epifluorescent microscopy (DEM) and swabbing and TVC analysis, beyond the three log reduction observed at 17.2 bar. The effect of spray time at 17.2 bar showed that increasing spray time from 1 to 10 s did not significantly increase removal of Ps. aeruginosa biofilm. Investigation of the optimum distance of the spray lance from the surface at 17.2 bar was found to be between 125 and 250 mm. The use of an alkaline, acidic or neutral detergent prior to spraying with water at 17.2 bar did not significantly increase the removal of Ps. aeruginosa or Staph. aureus. However, the acidic and alkaline products significantly (P = 0.05) affected the viability of Staph. aureus and Ps. aeruginosa, respectively, thereby minimizing the potential for the spread of contamination.     

Advantages of using microbial technology over traditional chemical technology in removal of black crusts from stone surfaces of historical monuments

This study compares two cleaning methods, one involving an ammonium carbonate-EDTA mixture and the other involving the sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris ATCC 29579, for the removal of black crust (containing gypsum) on marble of the Milan Cathedral (Italy). In contrast to the chemical cleaning method, the biological procedure resulted in more homogeneous removal of the surface deposits and preserved the patina noble under the black crust. Whereas both of the treatments converted gypsum to calcite, allowing consolidation, the chemical treatment also formed undesirable sodium sulfate.     

Comparing the bioremoval of black crusts on colored artistic lithotypes of the Cathedral of Florence with chemical and laser treatment

The external walls of the Cathedral of Florence are made of green serpentine, red marlstone and Carrara white marble, and intensive air pollution attack has led to their weathering, which caused black crust formation. A study was performed to evaluate the most appropriate cleaning treatment for black crust removal, adopting chemical (ammonium carbonate poultice), laser (1064 nm, Nd:YAG laser), and microbial (poultice embedding sulfate-reducing bacteria) cleaning. The effects of the different procedures on the original surfaces were evaluated by scanning electron microscopy coupled with energy dispersive X-ray (SEM/EDS) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and color measurements. One year later further color measurements were made. It was found that chemical cleaning led to non-homogeneous crust removal and that for the extremely powdery and incoherent red substratum the preferred treatment was laser cleaning. Overall, the most satisfactory treatment was the microbial cleaning process. It was the most controllable process and the most efficient for sulfate removal. Its main drawback appears to be the time needed to remove thick black crusts since numerous applications were necessary.     

Advantages of Using Microbial Technology over Traditional Chemical Technology in Removal of Black Crusts from Stone Surfaces of Historical Monuments

This study compares two cleaning methods, one involving an ammonium carbonate-EDTA mixture and the other involving the sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris ATCC 29579, for the removal of black crust (containing gypsum) on marble of the Milan Cathedral (Italy). In contrast to the chemical cleaning method, the biological procedure resulted in more homogeneous removal of the surface deposits and preserved the patina noble under the black crust. Whereas both of the treatments converted gypsum to calcite, allowing consolidation, the chemical treatment also formed undesirable sodium sulfate.     

Assessment of the potential suitability of selected commercially available enzymes for cleaning-in-place (CIP) in the dairy industry

The potential suitability of 10 commercial protease and lipase products for cleaning-in-place (CIP) application in the dairy industry was investigated on a laboratory scale. Assessment was based primarily on the ability of the enzymes to remove an experimentally generated milk fouling deposit from stainless steel (SS) panels. Three protease products were identified as being most suitable for this application on the basis of their cleaning performance at 40 °C, which was comparable to that of the commonly used cleaning agent, 1% NaOH at 60 °C. This was judged by quantification of residual organic matter and protein on the SS surface after cleaning and analysis by laser scanning confocal microscopy (LSCM). Enzyme activity was removed/inactivated under conditions simulating those normally undertaken after cleaning (rinsing with water, acid circulation, sanitation). Preliminary process-scale studies strongly suggest that enzyme-based CIP achieves satisfactory cleaning at an industrial scale. Cost analysis indicates that replacing caustic-based cleaning procedures with biodegradable enzymes operating at lower temperatures would be economically viable. Additional potential benefits include decreased energy and water consumption, improved safety, reduced waste generation, greater compatibility with wastewater treatment processes and a reduction in the environmental impact of the cleaning process.     

Assessment of the potential suitability of selected commercially available enzymes for cleaning-in-place (CIP) in the dairy industry

The potential suitability of 10 commercial protease and lipase products for cleaning-in-place (CIP) application in the dairy industry was investigated on a laboratory scale. Assessment was based primarily on the ability of the enzymes to remove an experimentally generated milk fouling deposit from stainless steel (SS) panels. Three protease products were identified as being most suitable for this application on the basis of their cleaning performance at 40°C, which was comparable to that of the commonly used cleaning agent, 1% NaOH at 60°C. This was judged by quantification of residual organic matter and protein on the SS surface after cleaning and analysis by laser scanning confocal microscopy (LSCM). Enzyme activity was removed/inactivated under conditions simulating those normally undertaken after cleaning (rinsing with water, acid circulation, sanitation). Preliminary process-scale studies strongly suggest that enzyme-based CIP achieves satisfactory cleaning at an industrial scale. Cost analysis indicates that replacing caustic-based cleaning procedures with biodegradable enzymes operating at lower temperatures would be economically viable. Additional potential benefits include decreased energy and water consumption, improved safety, reduced waste generation, greater compatibility with wastewater treatment processes and a reduction in the environmental impact of the cleaning process.     

Adhesion and removal kinetics of Bacillus cereus biofilms on Ni-PTFE modified stainless steel

Biofilm control remains a challenge to food safety. A well-studied non-fouling coating involves codeposition of polytetrafluoroethylene (PTFE) during electroless plating. This coating has been reported to reduce foulant build-up during pasteurization, but opportunities remain in demonstrating its efficacy in inhibiting biofilm formation. Herein, the initial adhesion, biofilm formation, and removal kinetics of Bacillus cereus on Ni-PTFE-modified stainless steel (SS) are characterized. Coatings lowered the surface energy of SS and reduced biofilm formation by > 2 log CFU cm^?2. Characterization of the kinetics of biofilm removal during cleaning demonstrated improved cleanability on the Ni-PTFE coated steel. There was no evidence of biofilm after cleaning by either solution on the Ni-PTFE coated steel, whereas more than 3 log and 1 log CFU cm^?2 of bacteria remained on the native steel after cleaning with water and an alkaline cleaner, respectively. This work demonstrates the potential application of Ni-PTFE non-fouling coatings on SS to improve food safety by reducing biofilm formation and improving the cleaning efficiency of food processing equipment.     

The development of an ultrasonic apparatus for the noninvasive and repeatable removal of fouling in food processing equipment

A new ultrasonic apparatus operating at a frequency of 40 kHz was developed to dislodge biofilms from food processing equipment in order to assess the effectiveness of cleaning protocols. Sonication conditions to remove biofilms and quantification by ATP-bioluminescence are described. An industrial meat process was developed at the laboratory level to form a biofilm with industrial characteristics. Our results show that the biofilm removal by sonication during 10 s is reproducible and four times greater compared to the swabbing method (83% removal of fouling material against 20%). Unlike the swabbing method, this ultrasonic apparatus permitted the immediately demonstration of the inefficiency (within 1 min) of an industrial meat cleaning protocol. This apparatus is portable, easy to use and can be operated by unskilled users.     

Residual particle free rough surfaces after rough blasting with steel grit in total hip arthroplasty]

INTRODUCTION: Several shot peening and grit blasting techniques are used to modify the surface in the production of hip endoprostheses. A number of publications in maxillo- facial- surgery and orthopedic surgery demonstrated significant contamination on alumina blasted surfaces. Latest research studies suspected an association between surface contamination and early failure of endoprostheses associated with third body wear. Additionally, the European standard EN 12010 requires surfaces free of residual particles. The aim of this study is to evaluate the effects of rough blasting using steel grit followed by a pH dependent cleaning procedures with regard to complying with EN 12010. MATERIAL AND METHOD: Ti6Al7Nb rods were rough blasted 1 time using steel grit ("Stahldrahtkorn", hardness: 56 HRC; R+K Draht GmbH, Leisnig). All blasting procedures were carried out with a special grit blasting device with direct pressure. One Ti6Al7Nb rod each was grit blasted with 3 and 4 bar pressure and 10 rods with 5 bar pressure. Surface roughness was measured by means of Mitutoyo- Surftest 301 three times before and after grit blasting. The mean value was calculated. A field emission scanning electron microscopy (SEM, LEO 1525) was used for the detection of residual particles on the surface of the rods with a backscattered electron detector. RESULTS: By rough blasting using steel grit a mean roughness of 54,9 microm and a maximum roughness of 61,4 microm could be achieved. The surface of all rods showed a considerable contamination with steel particles after grit blasting. After the a pH- dependent cleaning procedure a residual particle free surface was detected in all samples. DISCUSSION: The European standard EN 12010 can be fulfilled using rough blasting with steel grit followed by a pH dependent cleaning procedure.     

Towards an absolute scale for adhesion strength of ship hull microfouling

In-water ships’ hull cleaning enables significant fuel savings through removal of marine fouling from surfaces. However, cleaning may also shorten the lifetime of hull coatings, with a subsequent increase in the colonization and growth rate of fouling organisms. Deleterious effects of cleaning would be minimized by matching cleaning forces to the adhesion strength of the early stages of fouling, or microfouling. Calibrated waterjets are routinely used to compare different coatings in terms of the adhesion strength of microfouling. However, an absolute scale is lacking for translating such results into cleaning forces, which are of interest for the design and operation of hull cleaning devices. This paper discusses how such forces can be determined using computational fluid dynamics. Semi-empirical formulae are derived for forces under immersed waterjets, where the normal and tangential components of wall forces are given as functions of different flow parameters. Nozzle translation speed is identified as a parameter for future research, as this may affect cleaning efficacy.     

Evaluation of the effect of cleaning regimes on biofilms of thermophilic bacilli on stainless steel

AIMS: To determine the mechanism for both the removal and inactivation of 18-h biofilms of a thermophilic Bacillus species that optimally grows at 55 degrees C on stainless steel. METHODS AND RESULTS: The cleaning strategies tested were based on biofilm biochemistry and physiology, and focused on the chemistry of the cleaners, the duration and temperature of the cleaning process and a combination of various cleaners. The success of the cleaning regimes was determined based on the removal of cells and organic debris and the elimination of viable cells. The results confirmed that a caustic (75 degrees C for 30 min) and acid (75 degrees C for 30 min) wash, relied upon heavily in most food processing industries for cleaning-in-place systems, was successful in removing these biofilms. However, any changes in the concentrations of these cleaners or the temperature of cleaning drastically affected the overall outcome. Alternative cleaning agents based on enzymatic or nonenzymatic breakdown of cellular proteins or polysaccharides, surfactant action, use of oxidative attack and free radicals varied in degrees of their success. Combining proteolytic action with surfactants increased wetability and therefore enhanced the cleaning efficiency. CONCLUSIONS: Several procedures, including caustic/acid and enzyme based cleaners, will be satisfactory, provided that the correct process parameters are observed i.e. concentration, time, temperature and kinetic energy (flow). Confirmation of these results should be carried out in a pilot plant through several use/clean cycles. SIGNIFICANCE AND IMPACT OF THE STUDY: Confidence in standard and alternative cleaning procedures for food manufacturing plant to prevent contamination with thermophilic bacilli that threaten product quality.     

Cleaning efficacy of hydroxypropyl-beta-cyclodextrin for biofouling reduction on reverse osmosis membranes

In this study, an environmentally friendly compound, hydroxypropyl-beta-cyclodextrin (HP-β-CD) was applied to clean reverse osmosis (RO) membranes fouled by microorganisms. The cleaning with HP-β-CD removed the biofilm and resulted in a flux recovery ratio (FRR) of 102%. As cleaning efficiency is sometimes difficult to determine using flux recovery data alone, attached bacterial cells and extracellular polymeric substances (EPS) were quantified after cleaning the biofouled membrane with HP-β-CD. Membrane surface characterization using scanning electron microscopy (SEM), attenuated total reflectance Fourier transform infrared (ATR-FTIR) and atomic force microscopy (AFM) confirmed the effectiveness of HP-β-CD in removal of biofilm from the RO membrane surface. Finally, a comparative study was performed to investigate the competitiveness of HP-β-CD with other known cleaning agents such as sodium dodecyl sulfate (SDS), ethylenediaminetetraacetic acid (EDTA), Tween 20, rhamnolipid, nisin, and surfactin. In all cases, HP-β-CD was superior.     

Impact of inorganic salts on degradation of bisphenol A and diclofenac by crude extracellular enzyme from Pleurotus ostreatus

This study investigated the influence of inorganic salts on enzymatic activity and the removal of trace organic contaminants (TrOCs) by crude laccase from the white-rot fungus Pleurotus ostreatus. A systematic analysis of 15 cations and anions from common inorganic salts was presented. Laccase activity was not inhibited by monovalent cations (i.e. Na⁺, NH₄⁺, K⁺), while the presence of divalent and trivalent cations showed variable impact – from negligible to complete inhibition – of both laccase activity and its TrOC removal performance. Of interest was the observation of discrepancy between residual laccase activity and TrOC removal in the presence of some ions. Mg²⁺ had negligible impact on residual laccase activity but significant impact on TrOC removal. Conversely, F^? showed greater impact on residual laccase activity than on TrOC removal. This observation indicated different impacts of the interfering ions on the interaction between laccase and TrOCs as compared to that between laccase and the reagent used to measure its activity, implicating that residual laccase activity may not always be an accurate indicator of TrOC removal. The degree of impact of halides was in the order of F^??>?I^? >?Br^??>?Cl^?. Particularly, the tolerance of the tested laccase to Cl^? has important implications for a range of industrial applications.     

Facultative cleaning of spiral‐horned antelope by the African paradise flycatcher (Terpsiphone viridis)

In cleaning associations, individuals known as “cleaners” remove and feed on parasites and pests found on, or around, other animals known as “clients.” While best documented in marine environments and as mutualisms, cleaning associations are widespread in terrestrial systems and range along a spectrum of obligate to facultative associations. In African savannas, cleaning associations primarily comprise facultative interactions between mammals and birds that remove attached parasites. Few reports, however, exist on cleaning associations that involve the removal of unattached pests. In this short note, I report a novel facultative bird–ungulate cleaning association involving the removal of unattached pests, between the African paradise flycatcher (Terpsiphone viridis) and two species of spiral‐horned antelope (Tragelaphus spp.): greater kudu (Tragelaphus strepsiceros) and Cape bushbuck (Tragelaphus sylvaticus). On multiple occasions, I observed African paradise flycatchers hawking flying insects around greater kudu and a Cape bushbuck during the dry season at the Mpala Research Centre in Laikipia, Kenya. These observations document a rare feeding strategy for the African paradise flycatcher and are among the few records on cleaning interactions involving the removal of unattached pests.     

Achieving hygiene in the domestic kitchen: the effectiveness of commonly used cleaning procedures

AIMS: To quantify the transmission of Salmonella and Campylobacter to hands, cloths, and hand- and food-contact surfaces during the preparation of raw poultry in domestic kitchens, and to examine the impact on numbers of these bacteria of detergent-based cleaning alone, or in conjunction with thorough rising. METHODS AND RESULTS: Groups of volunteers prepared chickens for cooking. Surfaces were sampled either before cleaning or after cleaning using water and detergent with or without thorough rinsing. Although cleaning followed by rinsing consistently achieved decontamination of surfaces contaminated with Campylobacter, significant numbers of surfaces were still contaminated with low numbers of Salmonella. Where cloths contaminated with Salmonella were stored overnight, a reduction in the efficacy of detergent-based cleaning regimes was observed. CONCLUSIONS: Rinsing is the critical step in ensuring that bacteria are removed from surfaces during cleaning, but this may still leave residual contamination. Growth of Salmonella occurs in some contaminated cloths during overnight storage; Salmonella on cloths stored overnight are also more difficult to remove by washing. SIGNIFICANCE AND IMPACT OF THE STUDY: Rinsing, as part of the cleaning process, is a critical step in achieving hygiene in the kitchen. However, to achieve completely hygienic surfaces, the use of an antimicrobial agent may be necessary.     

Acoustic sensing and signal processing techniques for monitoring milk fouling cleaning operations

Large resource investments are necessary in order to minimize the limiting problems arising from food industrial intensive productivity. One of the most challenging concerns is the cleaning status uncertainty among heat transfer areas in dairy heat exchangers, since the effectiveness of this process cannot be easily validated. The present study aimed to develop a low‐power ultrasound sensing method for monitoring the removal of milk fouling deposits along cleaning processes inside an experimental plate heat exchanger structure, connected to a milk piping unit. For that purpose, signal processing, namely acoustic feature extraction, over different wave patterns combined with artificial neural network techniques was used. Measurements were taken in pulse‐echo mode with a handmade 4 MHz ultrasound transducer. While fouling deposits having initial average thickness values of 250 μm (34.5 ± 4.5 mg/cm²) were removed, the acoustic transmissivity increased. Results showed that the signal features follow the expected trends in both, clean and fouled cases, within right guess detection accuracies above 80%. Therefore, when calibrated well, this could be a very sensitive and noninvasive technique for material characterization, as well as a suitable validation method for industrial cleaning cycle operation optimization that could significantly reduce the associated costs.     

Modifications and additions to the ISO-DALT system for two-dimensional gel electrophoresis

Modifications of ISO-DALT devices that further enhance the efficiency and reproducibility of two-dimensional mapping of proteins are described. The principal changes in ISO system devices include the introduction of a gel casting trough with a removable panel to permit the removal of excess gel without introducing air into the electrofocusing gels and the introduction of an upper electrode compartment with a separate watertight septum for each electrofocusing tube to permit tube removal for cleaning and replacement. The principal changes in DALT system devices include the use of modified powder funnels to introduce acrylamide solutions into the slab gel gradient former without aeration; the introduction of a flexible outlet system for the gradient former to facilitate the removal of air bubbles; the introduction of an inexpensive two-part mixing chamber to permit disassembly for cleaning; the use of split gel holders to eliminate deformation and breakage of electrofocusing gels during loading onto slab gels; the introduction of an inexpensive integrated slab gel casting/rotating apparatus; and the introduction of a simple, water-cooled slab gel electrophoresis apparatus to reduce the volume of running buffer used in electrophoresis.