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.     

Ship hull in-water cleaning and its effects on fouling-control coatings

Abstract

Today, ship hull fouling is managed through fouling-control coatings, complemented with in-water cleaning. During cleaning, coating damage and wear must be avoided, for maximum coating lifetime and reduced antifoulant release. When possible, cleaning should target early stages of fouling, using minimal forces. However, such forces, and their effects on coatings, have not yet been fully quantified. In this one-year study, minimal cleaning forces were determined using a newly-designed immersed waterjet. The results show that bi-monthly/monthly cleaning, with maximum wall shear stress up to ～1.3?kPa and jet stagnation pressure ～0.17?MPa, did not appear to cause damage or wear on either the biocidal antifouling (AF) or the biocide-free foul-release (FR) coatings. The AF coating required bi-monthly cleanings to keep fouling to incipient slime (time-averaged results), while the FR coating had a similar fouling level even without cleaning. The reported forces may be used in matching cleaning parameters to the adhesion strength of the early stages of fouling.     

Fouling and its control in heat exchangers in the dairy industry

Fouling of heat transfer equipment poses a major problem in the food and especially the dairy industry. A review of recent progress made in the study and understanding of fouling and cleaning mechanisms in relation to exchangers and heat exchange surfaces in contact with milk components is given.

The first part describes the different materials and methods used in the study of fouling and cleaning, from laboratory to pilot‐plant and industrial scale.

The second part concerns the different fouling dynamics observed with milk and dairy products, and gives an interpretation of the different behaviours which occur, on the basis of the most recent results on the composition and structure of the deposits and the biochemical mechanisms of fouling.

The last part is a complete review of cleaning of milk deposits, taking into account the chemical reaction approach to cleaning as a mass transfer phenomenon.     

A comparative evaluation with respect to the bacterial cleanability of a range of wall and floor surface materials used in the food industry

J. H. TAYLOR AND J. T. HOLAH. 1996. A range of floor and wall materials suitable for use in the food industry were selected for comparative bacterial cleanability tests. A standard cleaning regime was carried out on samples after biofilm development of Acinetobacter calcoaceticus (CRA 296). Cleanability was measured as the log reduction in bacterial numbers. The results for the wall materials showed that there were no statistical differences, whilst those for the floor materials were more variable. The grouted joints and texture did not compromise the cleanabilities of tiled surfaces.     

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.     

In situ quantification of fish‐cage fouling by underwater photography and image analysis

Close‐up underwater photography and image analysis were used to quantify mesh occlusion by biofouling of salmon‐cage netting. This technique allows fast, non‐destructive sampling of cages in situ for the determination of temporal and spatial changes in fouling. The area of net blockage can be easily determined, allowing rapid evaluation of cleaning or antifouling performance.     

Rhamnolipid as new bio‐agent for cleaning of ultrafiltration membrane fouled by whey

In this work, rhamnolipid biosurfactant as an eco‐friendly and biodegradable cleaning agent was produced by Pseudomonas aeruginosa bacteria and was used to evaluate the chemical cleaning efficiency of whey fouled ultrafiltration membranes. Thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR) confirmed the successful synthesis of rhamnolipid. The produced rhamnolipid was compared to chemical cleaners including sodium hydroxide (NaOH), sodium dodecyl sulfate (SDS) and Tween 20. Ultrafiltration membranes used for fouling and cleaning analysis were prepared using phase inversion via immersion precipitation technique. For studying the fouling mechanisms, Hermia's model adapted to cross‐flow was used. From the fouling mechanism experiments, it was found that the complete blocking and cake formation were the dominant fouling mechanisms. The highest values of cleaning efficiency were achieved using rhamnolipid and NaOH as cleaning agents with the flux recovery of 100%, but with considering the low concentration of the rhamnolipid used in the cleaning solution compared to NaOH (0.3 versus 4 g/L for NaOH), its application is preferred.     

Evaluation of Brushes for Removal of Fouling from Fouling-release Surfaces,Using a Hydraulic Cleaning Device

A portable hydraulic device has been developed for use in optimizing the design of brushes and cleaning units that may be employed to maintain fouling-release coatings. Laboratory tests showed that characteristics of experimental cleaning brushes, including bristle stiffness, density, and angle, significantly affected the shear and normal forces imparted to the surface and thus, to any encrusting organisms. The standoff distance between the cleaning unit and the surface also influenced the forces generated. The rate of rotation of the brush, however, had little effect on force. The hydraulic device, with its experimental brushes, can also be used to evaluate the cleanability of fouling-release surfaces in situ, or to assess wear of the coating system due to cleaning.     

Commensalism or mutualism: conditional outcomes in a branchiobdellid–crayfish symbiosis

A growing body of evidence suggests that the costs and benefits involved in cleaning interactions can vary over space and time depending on the environmental conditions. However, whether the same cleaners actually induce variable net outcomes in terms of host fitness remains unclear to date. Branchiobdellid annelids are generally regarded as commensals or rarely parasites of their freshwater crayfish hosts, but a recent study suggests that they may also function as cleaning organisms. Under natural conditions, crayfish can experience fouling of the exposed surfaces of their exoskeletons and their gills (e.g., epibiosis) by various epibionts and particles of organic debris, and branchiobdellids graze on these sources of fouling. Here, we examined the extent to which variation in fouling pressure in the environment alters the outcome of the interaction between branchiobdellids and their crayfish host Cambaroide similis. A series of manipulations were performed in artificial environments designed to simulate either high or low fouling pressure. We used crayfish growth rates and mortality as direct measurements of the net costs and benefits of cleaning. Branchiobdellids had no significant effect on crayfish growth or mortality when cultured under low fouling pressure. However, their presence had a significant positive impact on host growth rates when cultured under high fouling pressure. These results suggest that the relationship between crayfish and branchiobdellids can fluctuate between commensalism and mutualism depending mainly on the environmental fouling pressure. We hypothesize that the outcome of cleaning interactions may largely depend on the factors directly related to the need for cleaning, such as parasite loads or fouling pressure.     

The prevention of microfouling and macrofouling on hydrogels impregnated with either Arquad 2C-75® or benzalkonium chloride

Abstract

Optically clear, surfactant loaded poly (2-hydroxyethyl methacrylate) (pHEMA) hydrogels can be used to prevent fouling on optical windows of marine underwater sensors. To act successfully in this capacity, hydrogels need to prevent both microfouling and macrofouling. Panel trials were conducted using four different materials: unloaded hydrogels, hydrogels containing either benzalkonium chloride (BAC) or dicocodimethylammonium chloride (Arquad 2C-75®) and PMMA coupons. Three panels were deployed at staggered intervals (2, 4 and 6 weeks) before the main settlement season of Semibalanus balanoidesand Mytilus edulisin the Firth of Clyde, Scotland. Panels were left for a total period of 10, 12 and 14 weeks respectively. Results showed that no sample completely resisted fouling, but Arquad 2C-75® hydrogels were extremely effective at preventing both microfouling and macrofouling. The most heavily fouled materials were unloaded hydrogels and PMMA, despite differences in initial hydrophilicities. Arquad 2C-75® hydrogels were equally effective at preventing larval settlement, for up to 14 weeks.     

Interspecific variation in patterns of adhesion of marine fouling to silicone surfaces

Abstract

The adhesion of six fouling organisms: the barnacle Balanus eburneus, the gastropod mollusc Crepidula fornicata, the bivalve molluscs Crassostrea virginica and Ostrea/Dendrostrea spp., and the serpulid tubeworms Hydroides dianthus and H. elegans, to 12 silicone fouling-release surfaces was examined. Removal stress (adhesion strength) varied among the fouling species and among the surfaces. Principal component analysis of the removal stress data revealed that the fouling species fell into two distinct groups, one comprising the bivalve molluscs and tubeworms, and the other the barnacle and the gastropod mollusc. None of the silicone materials generated a minimum in removal stress for all the organisms tested, although several surfaces produced low adhesion strengths for both groups of species. These results suggest that fouling-release materials do not rank (in terms of adhesion strength) identically for all fouling organisms, and thus development of a globally-effective hull coating will continue to require testing against a diversity of encrusting species.     

Antimicrobial and antifouling polymeric coating mitigates persistence of Pseudomonas aeruginosa biofilm

Abstract

Food wasted due to food spoilage remains a global challenge to the environmental sustainability and security of food supply. In food manufacturing, post-processing contamination of food can occur due to persistent bacterial biofilms, which can be resistant to conventional cleaning and sanitization. The objective was to characterize the efficacy of a polymeric coating in reducing Pseudomonas aeruginosa biofilm establishment and facilitating its removal. Viable cell density of a 48?h biofilm was reduced by 2.10 log cfu cm^?2 on the coated surface, compared to native polypropylene. Confocal laser scanning and electron microscopy indicated reductions in mature biofilm viability and thickness on the coated material. The antifouling coating improved cleanability, with ～2.5 log cfu cm^?2 of viable cells remaining after 105?min cleaning by water at 65?°C, compared to 4.5 log cfu cm^?2 remaining on native polypropylene. Such coatings may reduce the persistence of biofilms in food processing environments, in support of reducing food spoilage and waste     

麦田土壤真菌多样性对麦玉轮作长期秸秆还田和配施有机肥的响应

【背景】土壤真菌主导田间秸秆腐解的过程,秸秆还田配施有机肥可为真菌提供良好的营养物质。【目的】研究麦玉轮作模式下长期秸秆还田配施有机肥对麦田土壤真菌多样性的影响。【方法】依托山西南部麦玉轮作区长达14年的秸秆还田和施有机肥长期定位试验,采用高通量测序开展土壤真菌群落结构和多样性对不施肥+秸秆清茬(CK)、氮磷化肥+秸秆清茬(NP)、氮磷化肥+秸秆还田(SNP)、氮磷化肥+有机肥+秸秆清茬(NPM)及氮磷化肥+秸秆还田+有机肥(SNPM)的响应差异研究。【结果】秸秆还田和施有机肥处理组的物种丰富度指数、Chao1指数和ACE指数均高于CK。5个处理共产生953个分类操作单元(operational taxonomic units, OTU),CK、NP、NPM、SNP和SNPM分别具有398、451、472、462和440个OTU。在门水平上共检测出9个菌门,其中子囊菌门(Ascomycota)、毛霉门(Mucoromycota)和担子菌门(Basidiomycota)为3个主要菌门,各处理中所占丰度差异显著;在属水平上共检测出262个真菌属,其中CK丰度较高的3个属为柄孢壳菌属(Podospora) 18.85%、被孢霉属(Mortierella) 16.67%和镰刀菌属(Fusarium) 7.77%;NP、NPM、SNP和SNPM丰度较高的前3个相同属均为Dendrostilbella、毛葡孢属(Botryotrichum)和被孢霉属(Mortierella),但相对丰度值存在差异。由聚类分析可知,NPM和SNPM群落组成相似度高,归为一类,而CK、NP、SNP各独立为一类。与环境因子间冗余分析发现,TN含量是影响土壤真菌群落结构的关键因子,也受速效磷、pH、速效钾、碱解氮和全磷等环境指标影响。【结论】长期秸秆还田和施有机肥改变了小麦土壤真菌种群结构和多样性。     

川西亚高山针叶林树种云杉和冷杉土壤酸碱性差异及其机制

比较了川西亚高山针叶林主要树种粗枝云杉(Picea asperata)和岷江冷杉(Abies faxoniana)样地各土层(0—5 cm、5—10cm、10—20 cm)土壤pH值差异,并从两树种养分吸收和养分归还相关累积H~+输入方面探究其差异原因。研究结果表明:云杉样地各土层土壤pH值均显著高于冷杉样地(P0.05);云杉样地地被物累积H~+输入显著低于冷杉样地(P0.05);云杉样地凋落物中P、Mg、N、C平均浓度显著低于冷杉(P0.05),而Ca、C/N、木质素/N、C/P显著高于冷杉样地(P0.05),两树种凋落物中木质素、K平均浓度无显著差异。云杉和冷杉凋落物化学特性主成分分析PC_1、PC_2方差贡献率分别为73.7%和15.6%,累积方差贡献率为89.4%,其中PC_1主要综合Ca、C/P、C/N、木质素/N、P、N、Mg的信息;PC_2主要综合木质素、K、C的信息。各土层土壤pH值均与地被物累积H~+输入显著负相关,与PC_1样本分数显著正相关。研究结论:云杉和冷杉样地土壤pH值存在显著树种差异,且云杉使土壤pH值变大,冷杉使土壤pH值变小,这主要与地被物形成以及凋落物化学特性有关,即与凋落物的量和凋落物分解速率、凋落物养分归还率密切相关。     

Conditioning film formation and its influence on the initial adhesion and biofilm formation by a cyanobacterium on photobioreactor materials

Abstract

Although cyanobacteria are a common group of microorganisms well-suited to utilization in photobioreactors (PBRs), studies of cyanobacteria fouling and its prevention are scarce. Using a cyanobacterium, Anabaena sp. PCC 7120, which had been genetically modified to enhance linalool production, the formation of conditioning films and the effects of these on the physico-chemical surface properties of various PBR materials during initial adhesion and biofilm formation were investigated. The adhesion assay revealed that the overall attachment of Anabaena was substratum dependent and no correlation between the hydrophobicity/roughness of clean material and cell attachment was found. Surface hydrophilicity/hydrophobicity of all the materials changed within 12?h due to formation of conditioning films. ATR-FTIR spectroscopy revealed that the fractional change in protein deposition between 12 to 96?h was consistent with Anabaena cell attachment but polysaccharide deposition was material specific and did not correlate with cell attachment on the PBR materials. Also, the delay in conditioning film proteins on PVC and PTFE indicated that components other than proteins may be responsible for the decrease in contact angles on these surfaces within 12?h. This indicates the important role of the chemical nature of adsorbed conditioning films in determining the initial attachment of Anabaena to PBR materials. The lower rate of attachment of Anabaena on the hydrophilic surfaces (glass and PMMA) between 72?h to 96?h (regime 3) showed that these surfaces could potentially have low fouling characteristics at extended time scales and should be considered for further research.     

Cleaning characterization of protein drug products using UV-vis spectroscopy

This study uses on-line absorbance monitoring to evaluate cleanability of protein drug products. Characterization and validation of equipment cleanliness is a key requirement for a biopharmaceutical facility. A manufacturing-scale cleaning cycle has to be developed and validated for its ability to clean all of the equipment parts for a given soil. Cleaning validation in a multiproduct fill-finish facility could benefit from using a worst-case-based approach that involves validating the cleaning process for the most difficult to clean product. Such an approach minimizes the number of required validation runs. Scaled-down cleaning evaluations can provide helpful information for evaluating multiple products and determine the worst case. This study presents a simple and rapid technique for bench-scale characterization of cleanability of protein drug products. On-line A280 (UV absorbance at 280 nm) measurements are performed using a fiber optic probe, and the data are used to establish the dynamics of protein dissolution in cleaning solution. The model not only helps to estimate cleaning time of different formulated proteins (and peptides) but also provides insights into the kinetics of cleaning under different thermal and chemical conditions. Protein product degradation during cleaning is also evaluated through gel electrophoresis. Such information is useful in designing new cleaning cycles. While the study is performed using drug products, the model as well as the findings are also applicable for characterization of final purified bulk soils relevant to bulk drug manufacturing.     

A Microbiological Test Method to Determine the Cleanability of Filter Media in Solid‐Liquid‐Separation Applications

One of the major properties in the design and development of filter media for the food and drug industry is their cleanability. They should be designed according to hygienic criteria thereby decreasing residuals and reducing cleaning time and costs. A reproducible test method is necessary to determine or validate the degree of cleanability of filter media and to indicate areas of poor hygienic design. This work focused on the development of such a filter test and provides information about impurity, whether it occurs or not, and its local resolution. The growth of a microorganism, which is introduced as impurity, is used for the detection of residues which grow to the surface of the sample and offer a 2‐dimensional analysis. With this approach, different kinds of filter media can be compared with each other regarding their cleanability.     

阿尔泰山两河源自然保护区地面生地衣群落物种分布特征

用聚类分析和主成分分析法对阿尔泰山两河源自然保护区地面生地衣群落进行划分;根据优势地衣种类进行群落命名。应用典范对应分析法探讨了地面生地衣种类分布与环境间的关系。结果表明:阿尔泰山两河源自然保护区地面生地衣可分为以下4个群落:白腹地卷+硫石蕊+喇叭粉石蕊群落;茸珊瑚枝+长根地卷+黑瘿地卷群落;土星猫耳衣+软鹿蕊+膜地卷群落;雪岛衣+北方石蕊+绿皮地卷群落。各群落中的地面生地衣种类与盖度,在一定程度上反映了地面生地衣在阿尔泰山两河源保护区分布格局与环境的关系。典范对应分析结果表明,阿尔泰山地面生地衣群落物种分布受到土壤湿度、土壤pH、海拔高度、林冠层郁闭度和灌木层盖度等环境因素的影响,其中海拔、土壤湿度、林冠层郁闭度等对地衣物种分布的影响最大。     

The effect of wiping and spray-wash temperature on bacterial retention on abraded domestic sink surfaces

The relative cleanability of artificially abraded stainless steel, enamelled steel, mineral resin and polycarbonate domestic sinks was assessed by examining bacterial retention after cleaning. Two cleaning regimes were used: the mechanical action of wiping combined with a spray-rinse, and spray-washing at a range of temperatures. After wiping, stainless steel retained 0.5–1 log order fewer bacteria than the enamel sinks which in turn were 0.5 log order cleaner than the mineral resin and polycarbonate sinks. After spray-washing, stainless steel retained 0.5 log order fewer bacteria than enamel which in turn was 0.5 log order cleaner than the polycarbonate and mineral resin. Extending the number of wipes or increasing spray-wash temperature enhanced bacterial removal but, in general, did not change the relative cleanability of the sink materials. As a cleaning technique, wiping was shown to be more effective than spray-washing in reducing bacterial numbers. SEM studies showed that bacteria were typically retained in surface imperfections, particularly pits and crevices such that surfaces which sustained the most extensive damage due to abrasion retained higher numbers of bacteria.     

Influence of physicochemical properties on the hygienic status of stainless steel with various finishes

The relative hygienic status of 16 stainless steel surfaces, characterised by topography and surface free energy was investigated. B. thuringiensis spores suspended in Bechamel sauce was chosen as the test fouling suspension. Surface topography was assessed using 10 standardised roughness parameters, along with scanning electron microscope observations. The number of residual adhering spores after a fouling and cleaning in place procedure was found to be influenced by the topography of the stainless steel surface, but not by the surface free energy. Among the various roughness parameters, R_A, R_RR R_PK and R_VK were shown to be related to the hygienic status. Microscopic observations demonstrated the influence of the shape and size of surface irregularities on the level of residual soil after cleaning. This confirms that the use of only one roughness parameter, usually R_A, is not sufficient in defining the hygienic status of stainless steel surfaces.