This study characterised the bacterial flora of a commercially produced Stilton cheese in an effort to determine the contribution of non-starter lactic acid bacteria (NSLAB) to its aroma profile. A total of 123 microbial strains previously isolated from different sites (outer crust, blue veins and white core) of the cheese sample obtained at the end of ripening (~8 weeks) were recovered in MRS and BHI broths and preliminarily identified using conventional microbiological methods in order to establish population diversity and to screen out yeasts and moulds. Organisms identified with partial 16S rDNA sequence analysis were Lactobacillus plantarum, Lactobacillus brevis, Enterococcus faecalis, Staphylococcus aureus, Acinetobacter baumanii and Psychrobacter spp., with the genus Lactobacillus being the dominant (75%) group detected in all the sampled sites. Cluster analysis of pulse-field gel electrophoresis patterns associated the Lactobacillus isolates according to their site of isolation. Lb. plantarum isolates, two from each of the cheese sites, were evaluated for tolerance to heat stress and to different levels of salt, acid and relative humidity (RH) in order to ascertain whether the stress conditions associated with the isolation site could select the phenotype of microbial species recovered. The D72°C values revealed that isolates obtained from the outer crust were more heat sensitive suggesting they may have colonised the cheese post-pasteurisation. All the isolates were sensitive at pH range 3-4 but could grow at pH range 4.5-5. Similarly, isolates could grow at 3.5-5% (w/v) sodium chloride but were suppressed at 10%. Lactobacilli from the outer crust were the most halo-tolerant growing at 8% sodium chloride. For all strains, survival was low at 33-54% RH when cells were suspended in sterile de-ionised water but survived better at 33% RH in maximum recovery diluent (MRD) suggesting cellular protection by MRD. Lb. plantarum isolates from each site (outer crust=7; blue veins=19; white core=24) were tested for antimicrobial activity against Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Staph. aureus, Salmonella Typhimurium, Clostridium sporogenes, Lb. pentosus and Lactococcus lactis using the plate agar overlay and paper disc diffusion assays. All the 59 Lactobacillus isolates were tested for plantaricin EF genes using PCR. The nature of antimicrobial activity was examined using cell-free supernatants treated to neutralise acids and/or hydrogen peroxide. Treatment with proteinase K was used to ascertain whether activity was due to bacteriocin (putative plantaricin) production. On solid medium, the isolates had antimicrobial activity against Gram-negative and Gram-positive bacteria, each isolate showing activity against more than one species. Lb. pentosus, Ps. aeruginosa, E. coli and L. monocytogenes were the most sensitive whereas Cl. sporogenes was the most resistant spp. Activity against these organisms was mainly attributed to acid, and to a less extent, hydrogen peroxide and plantaricin production. Whereas Lb. plantarum isolates had a high prevalence of plantaricin EF genes, there was weak evidence for plantaricin production in liquid medium assays. Plantaricin production was only demonstrable among Lb. plantarum isolates from the veins and core against Lb. pentosus, implying the phenomenon was largely dependent on the genotype/strain of Lb. plantarum and was only active against closely related lactic acid bacteria. Subsequently, the effect of growth and survival dynamics of the different genotypes of the organism on the volatile aroma profiles of milk was examined. Individual isolates, one from each of the cheese sites, were co-cultured with acid-producing Lc. lactis (APL) and non acid-producing Lc. lactis (NAPL) in UHT milk under simulated cheese ripening conditions. During early fermentation (0-48 h, 30oC), the isolate obtained from the blue veins stimulated more growth of Lactococcus strains in mixed culture when compared to single cultures and to Lactobacillus isolates obtained from other sites in mixed culture. The volatile profiles of all Lb. plantarum strains grown alone were not significantly different (p>0.05). The type and levels of volatiles detected in mixed culture depended on the genotype/strain of Lb. plantarum inoculated as well as the acidification capability of Lc. lactis with which it was co-cultured. Co-culture of Lactobacillus isolates with APL resulted in increased aldehyde and alcohol production, whereas with NAPL only acetoin synthesis was enhanced. Salt addition had minimal effect on the volatile profiles. During further incubation (12 weeks, 18oC), growth of Lb. plantarum strains was better in salted samples inoculated with NAPL. The NAPL strain remained stable at 7 log10 CFU/ml throughout, while the APL rapidly declined from 9 to less than 5 log10 CFU/ml. The highest level of alcohols, organic acids and acetoin was detected from samples inoculated with the pure culture of the Lactobacillus isolate obtained from the blue veins. Co-culture of the isolate with APL enhanced acid and alcohol production, whereas its co-inoculation with NAPL increased acetoin synthesis. As Lb. plantarum is an incidental organism in cheese, its presence is unpredictable; it was therefore concluded that occurrence of different genotypes of the organism could be a major contributory factor to the variations in the cheese quality characteristics from batch to batch.

In the blue cheese Stilton the starter mould Penicillium roqueforti grows and sporulates during the ripening period and is considered to be responsible for the unique blue cheese aroma. However, the sporulation of the mould, which results in the formation of blue veins, takes place in a fraction of the Stilton matrix which overall is very heterogeneous. Most blue cheeses develop a secondary microflora of yeasts which may affect their aroma. The aim of this study was to investigate the yeast flora of Stilton, the aroma profile of the cheese and the role of the yeasts in the aroma production. The approach in this work was to study individually the different sections of Stilton (the blue veins, the white core and the outer crust) as previous studies have demonstrated each section has a differing bacterial flora. In addition to the classical microbiology, a series of molecular techniques (Denaturing Gradient Gel Electrophoresis, Restriction Fragment Length Polymorphism and Terminal RFLP) were compared and applied for the screening of the local fungal communities in the cheese. The results showed that the two approaches were complementary. It was concluded that the structure of the fungal community was different for each section of the cheese. The aroma profiles of the three different sections of Stilton were studied using solvent extraction Gas Chromatography-Mass Spectrometry (GC-MS), a headspace GC-MS technique (SPME GC-MS) and direct headspace analysis (Atmospheric Pressure Chemical Ionisation [APCI]-MS). The different sections of Stilton presented different aroma profiles. Overall, the blue and the outer crust had similar profiles. These two sections contained higher amount of ketones while the white contained higher amounts of alcohols and aldehydes. Yeast isolates and the starter Penicillium roqueforti were cultivated alone and in combination in a cheese model and the aroma production was studied with SPME GC-MS analysis. The co-culture of the starter Penicillium roqueforti and individual yeast isolates resulted in aroma profiles different from those that were produced by the mould or the yeasts individually. The model of Penicillium roqueforti with Yarrowia lipolytica resulted in an aroma more similar to blue cheese than produced by the mould alone. Sensory analysis (Flash profile technique) was used in order to compare the aroma of this model with the aroma of blue cheeses and the perception of the combined culture was found to be similar to Stilton cheese, whereas that of the mould alone was not. Yeasts are a significant part of the microflora of Stilton and they are able to affect the aroma production. Selected isolates of Yarrowia lipolytica could be used in combination with Penicillium roqueforti for the production of blue cheese aroma e.g. as a starter culture.

CCL3LI and CCL4LI are chemokine genes, located on chromosome 17q12. They are copy number variable genes which share 95% sequence identity with their non-copy number variable paralogues CCL3 and CCL4. The copy number of these genes varies between populations and has been reported to be associated with phenotypes such as susceptibility to HIV infection, hepatitis C virus infection, Kawasaki disease and SLE. The aim of this study is to understand the evolutionary history of variation at the CCL3L1/CCL4LI cluster. To accomplish this goal, several approaches including typing microsatellites, single nucleotide polymorphisms (SNPs) and CCL3L 1/CCL4L 1 sequence haplotypes were used to investigate the association with CCL3L 1 and CCL4L 1 copy number. However, the results showed that there is no strong association between a single-copy marker and CCL3L 1 and CCL4LI copy number, but there is evidence of recombination. Therefore, this may suggest that CCL3L 1/CCL4L 1 is a complex region and one plausible hypothesis is that there is a high rate of recombination in this region. This study of the evolution of CCL3L 1/CCL4L 1 haplotypes showed that a major one-copy CCL3L 1/CCL4L I haplotype (about 70% haplotype frequency) identified in humans, represents the ancestral state, as inferred from comparison with chimpanzee.

The overall aim of this study was to eliminate the root cause of the ‘cheese flavour’ in spread caused by four key microbes Bacillus spp, Staphylococcus spp, yeasts and moulds. The major sources of these bacteria were in the product ingredients mainly sweet cream buttermilk and skimmed milk along with environmental aerosols. The causative organisms were present in about 63% of the products and mainly ‘feed’ on the oil element of the recipe, containing high level C12 that generates the distinctive cheese flavour when broken down by bacteria. The key hurdle factor in spread preventing microbial growth is water droplet size. The spread showing cheese off flavour had a droplet size distribution of 95% <10 micron. To achieve finer droplet size distribution, trial products were made in the Scrape Surface Heat Exchanger (SSHE) over the current churning method with a distribution of droplet size 98% <5 micron. The trial product showed a 50% reduction in the generation of the ‘cheese flavour’ methyl ketones. The Staphylococcus spp cross contamination source where from personnel with direct food contact processing area. Further education on personal hygiene helped to reduce the level of Staphylococcus spp contamination in the product. The trial product from the SSHE was further challenge tested with Listeria monocytogenes over a 10 week shelf life period to evaluate product robustness against microbial growth and spoilage. The organism did not show any growth over the period of time. The liquid phase of the emulsion was further modified with various salts at different concentrations and challenged with L. monocytogenes isolated from various parts of the dairy environment. It was observed that a pH range of 5.5 or lower with added 0.063% potassium sorbate showed significant antibacterial affect compared to the nutrient enriched MPC-broth and the unsalted liquid phase of the emulsion with no added potassium sorbate. To understand L. monocytogenes survival within a dairy process, the organism was further challenged by exposure to pasteurisation heat treatments and the standard CIP cycle of acid and caustic treatment. No recovery rate of the organism was observed. Therefore it could be concluded that the contamination within the industry is more likely to be post process or environmental contamination rather than survival through the plant itself as per RASFF alert of Listeria spp outbreak in dairy. Therefore, reducing the available water in the liquid phase of the spread and achieving a <5 μm droplet size and a finer distribution within the product will be limiting factors to microbial growth. An air purifier system BAXX has reduced the level of environmental contaminants, especially yeast and mould.

The two main objectives in this study were investigating the microbial quality and microbial communities of 'use-by date' Mung bean sprouts by using conventional culture and 16S/18S rDNA PCR-DGGE methods, and evaluating the efficacy of natural antimicrobial substances, chemical disinfectants, and thermal treatments in reducing and inhibiting the growth of the pathogens on mung bean seeds. Retail samples of pre-packed mung bean sprouts were obtained from three retailers in the local area. The microbial quality and communities were evaluated on the 'use-by date'. The highest counts of total aerobic counts (7.86 log10 CFU/g), yeasts and moulds (7.0 log10 CFU/g), total lactic acid bacteria (6.24 log10 CFU/g) and total coliforms (6.63 log10 CFU/g) were found in samples from one shop and the DGGE band sequences also identified major populations of LAB from the same samples, These indicated poor quality and spoilage of the samples from this location and could be related to improper storage at temperatures above 5°C. The combination of conventional culture methods with the PCR-DGGE technique revealed a larger diversity of bacterial communities than eukaryotic ones based on the relative number of amplimers present on most of the OGGE gels. Identification based on band analysis revealed that the Enterobacteriaceae (29.6%), soil bacteria (20.4%), lactic acid bacteria (18.5%). yeast (14.8%). Pseudomonas spp. (13%), and Flavobacterium (3.7%) constituted the major populations in bean sprout samples. Cluster analysis of the OGGE patterns of both 16S and 18S rDNA amplimers found no strong relationship between sample sources and batches indicating the variability of natural populations. The use of natural antimicrobial products, such as a mixture of lime juice and vinegar (1: 1, pH 2.83) and bacteriocin-like substances produced by Pediococcus acidilactici, failed to reduce and inhibit the growth of Listeria monocytogenes on mung bean seeds. The former solution had higher antimicrobial efficiency in reducing the pathogen on seeds (1.93 log10 CFU/g) compared to the Pediococcus broth culture (1.22 log10 CFU/g), but both solutions failed to inhibit the re-growth of the pathogen during the sprouting process and also reduced seed germination percentage by 13-18%. The evaluation of efficacy of sequential washing using a combination of chemical treatments (two-step dipping) against the pathogens on seeds showed that a two-step dipping treatment in a solution containing 2% sodium hypochlorite for to min followed by 5% lactic acid solution for 5 min was the most effective treatment. This treatment achieved the highest reductions of L. monocytogenes (2.91 log10 CFU/g) and Salmonella Typhimurium (3.20 l0g10 CFU/g) after treatment and continued to reduce the pathogen during the sprouting process. This may be due to the chemical residues on treated seeds which lowered both pathogens on sprouted seeds to below the limit of detection <50 CFU/g) by direct plating without significantly affecting seed viability. The use of thermal treatments based on a hot and cold water dipping was found to be more effective in reducing the normal flora on seeds and less affecting of seed germination compared to microwave heating. The use of a hot and cold water dipping treatment at 92°c for 1 min followed by ice-cold water at 5°C for 30 sec achieved the highest reduction of L. monocytogenes on seeds (>5 log10 CFU/g) but had the lowest germination percentage (89%) compared to other hot and cold water dipping treatments. Microwave heating at 1-4 kW showed a poorer efficiency in reducing nonnal flora on seeds and severely affected seed viability. Overall, a two-step washing with 2% sodium hypochlorite followed by 5% lactic acid seems to be the most successful treatment in reducing and inhibiting the recovery of the pathogen during the sprouting process. However, the chemical residues on treated seed may become a negative image to apply this treatment in the sprout industry.

Blue Stilton is a blue-veined cheese manufactured in a restricted area of the UK, using lactic starter cultures plus a secondary culture of Penicillium roquefotti. The aim of this study was to determine the change in microflora during ripening of the cheese and to investigate potential microbial interactions. Additionally, the volatile compounds present in mature samples of cheeses exhibiting few blue veins were compared with those in good quality cheeses, showing ample blue veining. Experiments on cheeses from a single dairy, monitored during the ripening process, showed that the total Lactobacillus count increased from levels of around 103 cfu g-1 on day one to around 107 cfu g-1 after 8 weeks of ripening. This is comparable to values found in other cheeses including both mould-ripened and non mould-ripened varieties. Yeast counts were generally higher than those found in other cheeses and also increased to levels in the region of 107 g-1. The total viable count (TVC) decreased from around 109 g-1 initially, reflecting the presence of the starter bacteria, to 107 g.1, suggesting a decline in the starter bacteria similar to that found in other cheeses. Mature cheeses always exhibited similar numbers of microorganisms although the species varied between cheeses. High quality, mature, cheeses were compared with sub-standard cheeses from the same production site. The predominant species of lactobacilli in good quality cheeses were Lb. plantarum and Lb. curvatus, whereas in poor quality cheeses Lb. brevis predominated. This corresponded to the results of gas chromatography-olfactometry, which indicated the presence of fruity off flavours in poor quality cheeses. Several strains of these species were isolated, as indicated by differing capabilities in utilisation of a range of carbon sources. Yeast species also varied between good and poor quality cheeses with Candida sphaerlca and C. catenulata predominating in good cheeses and C. famata, C. lipolytica and C. catenulata also occurring in both good and poor quality samples. Strain differences were observed by the biochemical profiles and two strains of C. famata demonstrated inhibitory effects against P. roqueforti when incubated under anaerobiosis. It was concluded that these strains may affect the development of blue veins in Stilton cheese when maturation conditions encourage their proliferation. Comparisons were made between samples of cheeses from several Stilton producers and the results suggested that although the levels of the groups of microorganisms tested were similar, the species of lactobacilli and yeasts present were different. This suggests that the indigenous microflora may have a significant impact on the flavour of cheeses from individual production sites. It was concluded that the microflora of Blue Stilton cheese may have a significant impact on the quality of the product both in terms of flavour and the development of the blue veins.

This thesis describes research investigating the side-effects of pesticides on soil microbial biomass and microbial activity, with particular reference to two recently developed pesticides, a fungicide, epoxiconazole, and a herbicide, quinmerac. In a dose-responsee xperiment,a pplication of thesep esticidest o a sandy loam soil, at up to 10 and 20 times field rate, had no significant effect on soil microbial biomass C or ninhydrin-reactive N, over 84 days incubation. There was also no effect on soil respiration, except for the higher rate quinmerac-treated soil, which evolved 13% lessC02-Cthan the control. The rate of mineralisation of epoxiconazole and quinmerac, and their long-term effect on soil respiration, were measured in three contrasting soils: a sandy loam, a silty clay loam, and a clay soil, using 14C -labelled active ingredients. The kinetics of the pesticides' mineralisation were quite different, epoxiconazole being hyperbolic, while quinmerac was sigmoidal. The maximum amount of mineralisation of both pesticides occurred in the silty clay loam soil, which had the lowest microbial biomass content. The mineralisation of the pesticides was increased by the addition of ryegrass, with the greatest effect in the silty clay loam soil, probably because of the large ryegrass C: biomass C ratio. The mineralisation of epoxiconazole was affected by the ryegrass amendment much more than quinmerac. Further additions of the pesticides had no significant effect on soil respiration or pesticide mineralisation. The mineralisation of epoxiconazole and quimnerac was further investigated in the silty clay loam soil, using samples with different crop management histories, and the effects of ryegrass and glucose amendment. Pesticide mineralisation was shown to be related to the amount of soil microbial biomass, indicating that the difference in mineralisation rate between the three soil types above was not due to differences in their crop management, but innate differences in soil chemistry and microbiology. Ryegrass addition stimulated the mineralisation of epoxiconazole more than quinmerac, while the reverse was true for glucose, indicating that the pesticides were being degraded by two distinct fractions of the microbial biomass. The effects of long-term cumulative field application of the pesticides benomyl, chlorfenvinphos, aldicarb, triadimefon and glyphosate, on soil microbial biomass and mineralisation of soil organic matter were investigated. The addition of aldicarb consistently increased the microbial biomass, due to its beneficial effect on crop growth, but this effect was not reflected in the rate of organic matter mineralisation. However, in general, the continued application of these pesticides for up to 19 years, at slightly higher than the recommended rates, had very little effect on the soil microbial population. The effects of epoxiconazole and triadimefon on soil ergosterol content and microbial biomass C were compared in a sandy loam soil. Both pesticides temporarily reduced soil ergosterol by about 30%, while biomass C remained largely unaffected. However, when straw was added to the soils, the inhibition of ergosterol was still evident, as was an inhibitory effect on biomass C. The measurement of soil ergosterol was more sensitive to the pesticide effects than biomass C, and could be a useful test in determining changes in fungal populations.

Viruses have been shown to be responsible for considerable bacterial mortality and nutrient cycling in aquatic systems. As yet no detailed studies have been published on the role of viruses in natural soil bacterial communities despite common knowledge that viruses exist in the soil. This thesis sought to address some key questions on the ecology of soil bacterial viruses and their hosts. Disturbance through soil desiccation, nutrient inputs, rhizosphere effects and protozoan predation pressure were investigated. The first study of lysogeny in natural soil systems was also undertaken. The work presented here utilised tools and techniques commonplace in aquatic systems research and applied them to soil. A novel protocol was developed based on physical extraction of bacteria and viruses from soil and direct counting with epifluorescence microscopy. Physical extraction was achieved using shaking, ultrasound sonication and low speed centrifugation. The fluorochrome SYBR Gold was used to stain nucleic acid of extracted bacteria and viruses, and image analysis software used to determine bacterial cell volumes. Bacterial and viral abundances were in the region of 107-109 per gram of soil over a range of soil types. Significant fluctuations in viral and bacterial abundances were recorded at timescales of less than 24 h. Glucose and nitrogen addition led to substantial increases in bacterial and viral abundance. Loss of soil moisture resulted in peaks of viral abundance in sandy soils but not in a clay soil. A six-week microcosm study demonstrated that phage were not a significant regulator of bacterial abundance. Low levels of lysogeny were recorded over a range of soils when measured explicitly with Mitomycin C. The implication from that study was that viruses in soil behave differently to those in aquatic systems. Bacterial and viral abundances were highly coupled in most instances, irrespective of the potential activity of bacteria. Further fundamental studies are recommended.

Stichelton is a blue-veined raw milk cheese which is made following the Stilton cheese making process. In a previous study, a preliminary examination of its microflora during production was examined by traditional culture methods and initial PCR DGGE profiling. The aim of this study was to complete the profiling of Stichelton cheese and examine the contribution of its microbiota components to product characteristics. Stored samples of cheese production isolates were sequenced and in addition whole population DNA was extracted directly from a fully ripened Stichelton cheese (12 weeks) and from bulk cell suspensions collected on various media. The V3, V4V5 and V6V8 regions of 16S rDNA were amplified by PCR and separated by DGGE using 20 – 80 % urea formamide denaturing gradient. While Lactobacillus casei/paracasei, Staphylococcus equorum, Bacillus sp., Brevibacterium sp., Halomonas sp., Acinetobacter sp., Alkalibacterium sp. and Corynebacterium casei were only found by the molecular method, traditional culture detected a large number of potentially raw milk microbiota. Lactococcus lactis was detected in the raw milk sample and along the process by both methods. The L. lactis subsp. lactis which was detected in the core of matured Stichelton was shown by PFGE to be from the raw milk and not the starter culture used in Stichelton production. S. equorum was found in the crust of cheese pre-piercing and in all parts of the cheese post-piercing by the molecular approach only. This suggested this organism was introduced originally via handling. Five S. equorum isolated from Stilton, Danish Blue and Reblochon could grow up to 10% salt but did not tolerate low pH levels suggesting S. equorum in Stichelton must have been introduced by handling before or during ripening as if it was present in the early fermentation then it would die as fermentation progressed due to pH sensitivity. A model cheese system made with commercial UHT milk was used to examine the interaction between mixed Lc. lactis, P. roqueforti and S. equorum B2 isolated from the Stilton crust. S. equorum was either added 1.5 h after the addition of L. lactis or it was smeared on the surface of the cheese immediately after un-moulding. The viable counts and pH were analysed throughout the process, while texture, water activity and flavour volatiles using GCMS SPME were determined for one month ripened cheeses only. The results showed S. equorum survived in the cheese following either method of introduction and that in cheeses without P. roqueforti addition, the presence of incorporated and surface-spread S. equorum could inhibit the surface growth of a contaminant Penicillium. It also slowed the growth of starter P. roqueforti in cheeses made with this mould. A paler coloured crust, firmer textured cheese and a low amount of alcohols were shown in the model cheeses made with surface-smear S. equorum. Conversely, addition of S. equorum in the initial process made the cheese core softer and produced low amounts of acids. Ethanol, 3-methyl-1-butanol and 2-pentanone were the main flavour compounds in the model cheeses examined. The antifungal activity of the isolate was confirmed in laboratory media. Its ability to prevent Penicillium surface growth could be beneficial for white cheeses where this is an undesirable flaw. The results showed that the sporulation inhibitory effect on P. roqueforti was because of an antifungal agent produced by S. equorum, but it was not acid, bacteriocin or H2O2. Further study is needed to detect the antifungal agent. Overall, the study has expanded the understanding of the role non-starter bacteria may have in contributing to cheese ripening.

Exposure of artificial surfaces such as ship hulls to a marine environment leads to the attachment of assorted biomolecules, single celled organisms and marine invertebrates such as barnacles or mussels. Together, they form a structure known as a biofilm. These films lead to higher fuel consumption and add considerable expense to the operation of ships used by industrial and naval organisations. The work presented in this thesis describes the surface analysis of a novel poly(dimethylsiloxane) (PDMS) based foul-release coating. The coating also contains poly(ethylene glycol) groups (PEG). The differing chemical properties between these two domains led to an observed surface modification effect in water, whereby contact angle measurements decreased from ~110o to ~65 o over a period of five minutes. This effect was rapidly reversible on drying. Time of Flight-Secondary Ion Mass Spectrometry cryogenic depth profiling experiments confirmed this change in surface chemistry where the frozen surface of the coating was shown to have a higher intensity of ions associated with PEG groups at the surface compared to that in the bulk. Water immersion also led to a swelling of the surface seen by a change in the surface topography by Atomic Force Microscopy investigations. When applied to glass surfaces the coatings were flat and generally defect free regardless of the application method used. On exposure to Pseudomonas aeruginosa the coatings were found to be ten times more effective at preventing bacterial adhesion in the first instance than a PDMS standard. The mechanism of action was shown to be non-toxic by live/dead staining and did not appear to affect the way in which bacteria move on a surface. A flow adhesion assay demonstrated that a flow rate of almost two orders of magnitude lower was required to remove fifty percent of bacteria from a coated surface than on a glass standard, demonstrating the foul-release ability of the switching coating. Sea trials in a French coastal region highlighted the importance of exposing candidate coatings to a true marine environment for a suitable duration in order to determine their potential for use. Ultimately we show that the coating presented is a candidate for use as an effective coating for preventing marine biofouling and surface analysis was deemed to be an appropriate methodology to analyse coatings that have changing properties on exposure to water.