Evaluation of PTR-ToF-MS as a tool to track the behavior of hop-derived compounds during the fermentation of beer.

Hop-derived volatile organic compounds (VOCs) play an important role in the flavor and aroma of beer, despite making up a small percentage of the overall profile. To understand the changes happening during fermentation, proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS) was applied for the first time in brewing science to directly measure the changes in hop-derived VOCs during the fermentation of four different worts containing one of two aroma hops in combination with one of two yeast biotypes. PTR-ToF-MS successfully detected and tracked mass-to-charge ratios (m/z) arising from interactions between the different yeast strains and the hop cultivars. Differences were observed in the dynamic VOC profiles between different beer treatments for m/z such as m/z 145.121 (ethyl hexanoate) and m/z 173.153 (isoamyl isovalerate or ethyl octanoate). The ability to monitor changes in VOCs during fermentation provides valuable information on the priority of production and transformation reactions by yeast.

Effect of medium compositions on microbially mediated volatile organic compounds release profile.

AIMS: To monitor temporal changes in the volatile organic compounds' (VOCs) profile generated by the metabolic activities of Pseudomonads in real time. METHODS AND RESULTS: Three Pseudomonas strains were cultivated in Vogel's broth, supplemented with glucose (0·5 or 1%) and/or protein (egg white powder at 0 or 2%) at 25°C. Glucose or egg white protein contents influenced the VOCs' release profile for alcohols, carbonyls and sulphur derivatives. Increasing glucose content resulted in higher alcohol and ketone contents. Glucose showed a lower effect on the VOCs' release profile, mainly impacting on individual compounds, such as m/z 89 (3-methyl-1-butanol). In contrast, egg white protein enhanced production of VOCs such as m/z 75 (2-methyl-1-propanol) and m/z 63 (dimethyl sulphide) regardless of glucose level present in the medium. At the end of bacteria growth phase (54, 60 and 72 h), the fingerprint of VOCs was different from the early growth phase. Cells near to the end of their growth phase (54, 60 and 72 h) produced a distinctly different array of compounds compared to those produced in early growth phase, for example, cyclic compounds were detected in early growth phase, whereas sulphur derivatives were more common in late growth phase. CONCLUSIONS: Pseudomonads-mediated VOCs' fingerprint as a response to varying growth conditions can be identified as latent biomarkers. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding how microbially mediated VOCs' release profile responds to varying growth conditions can potentially be used as a rapid method for detecting microbial activities in controlled conditions such as food quality systems.

Understanding the effect of pulsed electric fields on thermostability of connective tissue isolated from beef pectoralis muscle using a model system.

Brisket is a low value/tough meat cut that contains a large amount of connective tissue. Conversion of collagen into gelatin during heating reduces the toughness of the connective tissue however this conversion is slow at low cooking temperatures (around 60°C). The objective of this project was to determine the ability of pulsed electric field (PEF) processing to reduce the thermal stability of connective tissue. To achieve this, a novel model system was designed in which connective tissue obtained from beef deep pectotalis muscle (brisket) was exposed to PEF at combinations of electric field strength (1.0 and 1.5kV/cm) and specific energy (50 and 100kJ/kg) within an agar matrix at electrical conductivities representing the electrical conductivity found in brisket. Differential scanning calorimetry showed that PEF treatment significantly (p<0.05) decreased the denaturation temperature of connective tissue compared to untreated samples. Increasing electric field strength and the specific energy increased the Ringer soluble collagen fraction. PEF treated samples showed higher solubilization compared to the untreated samples at both 60°C and 70°C in heat solubility test. SEM examination of PEF treated (at 1.5kV/cm and 100kJ/kg) and untreated samples revealed that PEF appeared to increase the porosity of the connective tissue structure. These finding suggest that PEF processing is a technology that could be used to improve the tenderness and decrease the cooking time of collagen rich, meat cuts.

PTR-MS volatile profiling of Pinot Noir wines for the investigation of differences based on vineyard site.

This study investigated the effect of vineyard site on the volatile profiles of Pinot Noir wines using proton-transfer reaction mass spectrometry with prior headspace dilution. The ANOVA and PCA enabled discrimination of wine based on vineyard site. Sample separation was due to differences in the ratios of a mixture of compounds, including higher alcohols, ethyl, and acetate esters. Proton-transfer reaction mass spectrometry appears to be a useful technique for rapidly discriminating wines based on vineyard site. The similarities and differences expressed in the wines' volatile profiles may help winemakers to reveal the potential of individual vineyard sites to produce wines of certain character. Copyright © 2017 John Wiley & Sons, Ltd.

Effective active vaccination against methamphetamine in female rats.

BACKGROUND: Immunotherapies directed against methamphetamine (MA) abuse have shown success in rodent models, however only a limited number of studies have investigated active vaccination in female mice and none in female rats. It is critical to determine if potential immunotherapeutic strategies generalize across sex, particularly for drugs that may produce significant sex-differences on behavioral or physiological endpoints. METHODS: Female Wistar rats were initially vaccinated with keyhole-limpet hemocyanin (KLH) or an anti-methamphetamine-KLH conjugate (MH6-KLH) three times over five weeks and implanted with radiotelemetry devices to assess locomotor activity and body temperature responses to MA. Rats were first exposed to MA via vapor inhalation (100mg/mL in propylene glycol) and then by injection (0.25-1.0mg/kg, i.p.) and vapor after a final vaccine boost. RESULTS: The MH6-KLH vaccine generated an increase in antibody titers across the initial 6-week, 3 immunization protocol and a restoration of titer after a week 14 booster. Locomotor stimulation induced by 0.25mg/kg MA, i.p, in the KLH group was prevented in the MH6-KLH group. MH6-KLH animals also exhibited an attenuated locomotor stimulation produced by 0.5mg/kg MA, i.p. No group differences in locomotion induced by vapor inhalation of MA were observed and body temperature was not differentially affected by MA across the groups, most likely because vapor inhalation of MA that produced similar locomotor stimulation resulted in ∼10-fold higher plasma MA levels. CONCLUSIONS: This study confirms the efficacy of the MH6-KLH vaccine in attenuating the effects of MA in female rats.

Critical analysis of the maximum non inhibitory concentration (MNIC) method in quantifying sub-lethal injury in Saccharomyces cerevisiae cells exposed to either thermal or pulsed electric field treatments.

Sub-lethal injury within a microbial population, due to processing treatments or environmental stress, is often assessed as the difference in the number of cells recovered on non-selective media compared to numbers recovered on a "selective media" containing a predetermined maximum non-inhibitory concentration (MNIC) of a selective agent. However, as knowledge of cell metabolic response to injury, population diversity and dynamics increased, the rationale behind the conventional approach of quantifying sub-lethal injury must be scrutinized further. This study reassessed the methodology used to quantify sub-lethal injury for Saccharomyces cerevisiae cells (≈ 4.75 Log CFU/mL) exposed to either a mild thermal (45°C for 0, 10 and 20min) or a mild pulsed electric field treatment (field strengths of 8.0-9.0kV/cm and energy levels of 8, 14 and 21kJ/kg). Treated cells were plated onto either Yeast Malt agar (YM) or YM containing NaCl, as a selective agent at 5-15% in 1% increments. The impact of sub-lethal stress due to initial processing, the stress due to selective agents in the plating media, and the subsequent variation of inhibition following the treatments was assessed based on the CFU count (cell numbers). ANOVA and a generalised least squares model indicated significant effects of media, treatments, and their interaction effects (P<0.05) on cell numbers. It was shown that the concentration of the selective agent used dictated the extent of sub-lethal injury recorded owing to the interaction effects of the selective component (NaCl) in the recovery media. Our findings highlight a potential common misunderstanding on how culture conditions impact on sub-lethal injury. Interestingly for S. cerevisiae cells the number of cells recovered at different NaCl concentrations in the media appears to provide valuable information about the mode of injury, the comparative efficacy of different processing regimes and the inherent degree of resistance within a population. This approach may provide similar information for other micro-organisms.

The effect of alginate lyase on the gentamicin resistance of Pseudomonas aeruginosa in mucoid biofilms.

AIMS: Pseudomonas aeruginosa can secrete large amounts of alginate during chronic infections and this has been associated with high resistance to antibiotics. The major aim of this study was to investigate whether degradation of extracellular alginate by alginate lyase would increase the sensitivity of Ps. aeruginosa to gentamicin, an aminoglycoside antibiotic. METHODS AND RESULTS: Degradation of alginate from Ps. aeruginosa was monitored using a spectrometric assay. Alginate lyase depolymerized alginate, but calcium and zinc cations at concentrations found in the cystic fibrosis lung reduced enzyme activity. Biofilms formed on agar were partially degraded by alginate lyase, but staining with crystal violet showed that the biomass of biofilms grown in liquid was not significantly affected by the enzyme. Viability testing showed that the sensitivity to gentamicin of biofilm bacteria and of bacteria released from biofilms was unaffected by alginate lyase. CONCLUSIONS: Our results show that at least under the conditions used here alginate lyase does not affect gentamicin resistance of Ps. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study indicates that alginate does not contribute to resistance to gentamicin and so does not provide support for the concept of treating patients with alginate lyase in order to increase the antibiotic sensitivity of Ps. aeruginosa.

Lipid tucaresol as an adjuvant for methamphetamine vaccine development.

The immunopotentiator tucaresol was modified for incorporation into liposomes, where it was found to be a superior adjuvant to MPLA for vaccination against methamphetamine.

Prediction of the amount and rate of histamine degradation by diamine oxidase (DAO).

Histamine is a biogenic amine that forms in a variety of foods and can cause food poisoning at high concentrations (>500 ppm). In situations where the formation of histamine in food cannot be prevented through refrigeration, diamine oxidase (DAO) enzyme may be used to degrade histamine to safe levels. The aims of this work were to apply DAO in model (buffer) and real (cooked tuna soup used in the manufacture of a fish paste product, Rihaakuru) systems, in order to obtain predictions for the rates and amounts of histamine degradation. The two systems were set up with a constant concentration of histamine (500 mg/L) and the DAO enzyme (2534 units/L) at a temperature of 37°C, agitation at 100 rpm and an incubation time of 10h with variable pH (5-7) and salt concentrations (1-5%). A total of 15 experiments were designed for each system using central composite design (CCD). The data from these experiments were fitted into regression models; initially the data were used to generate an exponential decline model and then the data from this were fitted into a secondary response surface model (RSM) to predict the rate and amount of histamine degradation by DAO. The model system results indicated that DAO activity was not significantly affected by salt (p>0.05), and that activity reached a maximum within the pH range of 6-6.5 with an optimum at pH 6.3. However, the results obtained with the tuna soup model showed that the optimum oxidation of histamine using DAO occurred between pH 6-7 and salt 1-3%. This study defined the conditions for the use of DAO to degrade 500 mg/L of histamine in tuna soup used to manufacture Rihaakuru. The models generated could also be used to predict the rate and amount of histamine degradation in other foods that have similar characteristics to tuna soup.

Diverse spore rains and limited local exchange shape fern genetic diversity in a recently created habitat colonized by long-distance dispersal.

BACKGROUND AND AIMS: Populations established by long-distance colonization are expected to show low levels of genetic variation per population, but strong genetic differentiation among populations. Whether isolated populations indeed show this genetic signature of isolation depends on the amount and diversity of diaspores arriving by long-distance dispersal, and time since colonization. For ferns, however, reliable estimates of long-distance dispersal rates remain largely unknown, and previous studies on fern population genetics often sampled older or non-isolated populations. Young populations in recent, disjunct habitats form a useful study system to improve our understanding of the genetic impact of long-distance dispersal. METHODS: Microsatellite markers were used to analyse the amount and distribution of genetic diversity in young populations of four widespread calcicole ferns (Asplenium scolopendrium, diploid; Asplenium trichomanes subsp. quadrivalens, tetraploid; Polystichum setiferum, diploid; and Polystichum aculeatum, tetraploid), which are rare in The Netherlands but established multiple populations in a forest (the Kuinderbos) on recently reclaimed Dutch polder land following long-distance dispersal. Reference samples from populations throughout Europe were used to assess how much of the existing variation was already present in the Kuinderbos. KEY RESULTS: A large part of the Dutch and European genetic diversity in all four species was already found in the Kuinderbos. This diversity was strongly partitioned among populations. Most populations showed low genetic variation and high inbreeding coefficients, and were assigned to single, unique gene pools in cluster analyses. Evidence for interpopulational gene flow was low, except for the most abundant species. CONCLUSIONS: The results show that all four species, diploids as well as polyploids, were capable of frequent long-distance colonization via single-spore establishment. This indicates that even isolated habitats receive dense and diverse spore rains, including genotypes capable of self-fertilization. Limited gene flow may conserve the genetic signature of multiple long-distance colonization events for several decades.

Computing Morse-Smale Complexes with Accurate Geometry.

Topological techniques have proven highly successful in analyzing and visualizing scientific data. As a result, significant efforts have been made to compute structures like the Morse-Smale complex as robustly and efficiently as possible. However, the resulting algorithms, while topologically consistent, often produce incorrect connectivity as well as poor geometry. These problems may compromise or even invalidate any subsequent analysis. Moreover, such techniques may fail to improve even when the resolution of the domain mesh is increased, thus producing potentially incorrect results even for highly resolved functions. To address these problems we introduce two new algorithms: (i) a randomized algorithm to compute the discrete gradient of a scalar field that converges under refinement; and (ii) a deterministic variant which directly computes accurate geometry and thus correct connectivity of the MS complex. The first algorithm converges in the sense that on average it produces the correct result and its standard deviation approaches zero with increasing mesh resolution. The second algorithm uses two ordered traversals of the function to integrate the probabilities of the first to extract correct (near optimal) geometry and connectivity. We present an extensive empirical study using both synthetic and real-world data and demonstrates the advantages of our algorithms in comparison with several popular approaches.

Effect of NaOH (caustic wash) on the viability, surface characteristics and adhesion of spores of a Geobacillus sp. isolated from a milk powder production line.

AIM: To investigate the viability, surface characteristics and ability of spores of a Geobacillus sp. isolated from a milk powder production line to adhere to stainless steel surfaces before and after a caustic (NaOH) wash used in clean-in-place regimes. METHODS AND RESULTS: Exposing sessile spores to 1% NaOH at 65°C for 30min decreased spore viability by two orders of magnitude. The zeta potential of the caustic treated spores decreased from -20 to -32 mV and they became more hydrophobic. Transmission electron microscopy revealed that caustic treated spores contained breaks in their spore coat. Under flow conditions, caustic treated spores suspended in 0·1 mol l(-1) KCl were shown to attach to stainless steel in significantly greater numbers (4·6 log(10) CFU cm(-2) ) than untreated spores (3·6 log(10) CFU cm(-2) ). CONCLUSIONS: This research suggests that spores surviving a caustic wash will have a greater propensity to attach to stainless steel surfaces. SIGNIFICANCE OF STUDY: The practice of recycling caustic wash solutions may increase the risk of contaminating dairy processing surfaces with spores.

Characterization of spore surfaces from a Geobacillus sp. isolate by pH dependence of surface charge and infrared spectra.

AIMS: The surfaces of spores from a Geobacillus sp. isolated from a milk powder production line were examined to obtain fundamental information relevant to bacterial spore adhesion to materials. MATERIALS AND RESULTS: The surfaces of spores were characterized using transmission electron microscopy and infrared spectroscopy. Thin sections of spores stained with ruthenium red revealed an exosporium with a hair-like nap around the spores. Attenuated total reflection infrared spectra of the spores exposed to different pH solutions on a ZnSe prism revealed that pH-sensitive carboxyl and phosphodiester groups associated with proteins and polysaccharides contributed to the spore's negative charge which was revealed by our previous zeta potential measurements on the spores. Lowering the pH to the isoelectric point of spores resulted in an increase in intensity of all spectral bands, indicating that the spores moved closer to the zinc selenide (ZnSe) surface as the charged surface groups were neutralized and the spore surface polymers compressed. The attachment of spores to stainless steel was threefold higher at pH 3 compared with pH 7. CONCLUSIONS: This research showed that spore attachment to surfaces is influenced by electrostatic interactions, surface polymer conformation and associated steric interactions. SIGNIFICANCE AND IMPACT OF THE STUDY: The adhesion of thermophilic spores is largely controlled by functional groups of surface polymers and polymer conformation.

Mechanisms of Cation Exchange by Pseudomonas aeruginosa PAO1 and PAO1 wbpL, a Strain with a Truncated Lipopolysaccharide.

The ability of bacterial cells to sequester cations is well recognized, despite the fact that the specific binding sites and mechanistic details of the process are not well understood. To address these questions, the cation-exchange behavior of Pseudomonas aeruginosa PAO1 cells with a truncated lipopolysaccharide (LPS) (PAO1 wbpL) and cells further modified by growth in a magnesium-deficient medium (PAO1 wbpL - Mg(2+)) were compared with that of wild-type P. aeruginosa PAO1 cells. P. aeruginosa PAO1 cells had a negative surface charge (zeta potential) between pH 11 and 2.2, due to carboxylate groups present in the B-band LPS. The net charge on PAO1 wbpL cells was increasingly positive below pH 3.5, due to the influence of NH(3)(+) groups in the core LPS. The zeta potentials of these cells were also measured in Na(+), Ca(2+), and La(3+) electrolytes. Cells in the La(3+) electrolyte had a positive zeta potential at all pH values tested. Growing P. aeruginosa PAO1 wbpL in magnesium-deficient medium (PAO1 wbpL - Mg(2+)) resulted in an increase in its zeta potential in the pH range from 3.0 to 6.5. In cation-exchange experiments carried out at neutral pH with either P. aeruginosa PAO1 or PAO1 wbpL, the concentration of bound Ca(2+) was found to decrease as the pH was reduced from 7.0 to 3.5. At pH 3.5, the bound Mg(2+) concentration decreased sharply, revealing the activity of surface sites for cation exchange and their pH dependence. Infrared spectroscopy of attached biofilms suggested that carboxylate and phosphomonoester functional groups within the core LPS are involved in cation exchange.

Recovery of spores from thermophilic dairy bacilli and effects of their surface characteristics on attachment to different surfaces.

Spores from four Geobacillus spp. were isolated from a milk powder manufacturing line in New Zealand. Liquid sporulation media produced spore yields of approximately 10(7) spores ml(-1); spores were purified using a two-phase system created with polyethylene glycol 4000 and 3 M phosphate buffer. The zeta potentials of the spores from the four isolates ranged from -10 to -20 mV at neutral pH, with an isoelectric point between pH 3 and 4. Through contact angle measurements, spores were found to be hydrophilic and had relative hydrophobicity values of 10 to 40%, as measured by the microbial adhesion to hexadecane assay. The most hydrophilic spore isolate with the smallest negative charge attached in the highest numbers to Thermanox and stainless steel (1 x 10(4) spores cm(-2)), with fewer spores attaching to glass (3 x 10(3) spores cm(-2)). However, spores produced by the other three strains attached in similar numbers (P > 0.05) to all substrata (approximately 1 x 10(3) spores cm(-2)), indicating that there was no simple relationship between individual physicochemical interactions and spore adherence. Therefore, surface modifications which limit the attachment of one strain may not be effective for all stains, and control regimens need to be devised with reference to the characteristics of the particular strains of concern.

Understanding the structure of the turbulent mixing layer in hydrodynamic instabilities.

When a heavy fluid is placed above a light fluid, tiny vertical perturbations in the interface create a characteristic structure of rising bubbles and falling spikes known as Rayleigh-Taylor instability. Rayleigh-Taylor instabilities have received much attention over the past half-century because of their importance in understanding many natural and man-made phenomena, ranging from the rate of formation of heavy elements in supernovae to the design of capsules for Inertial Confinement Fusion. We present a new approach to analyze Rayleigh-Taylor instabilities in which we extract a hierarchical segmentation of the mixing envelope surface to identify bubbles and analyze analogous segmentations of fields on the original interface plane. We compute meaningful statistical information that reveals the evolution of topological features and corroborates the observations made by scientists. We also use geometric tracking to follow the evolution of single bubbles and highlight merge/split events leading to the formation of the large and complex structures characteristic of the later stages. In particular we (i) Provide a formal definition of a bubble; (ii) Segment the envelope surface to identify bubbles; (iii) Provide a multi-scale analysis technique to produce statistical measures of bubble growth; (iv) Correlate bubble measurements with analysis of fields on the interface plane; (v) Track the evolution of individual bubbles over time. Our approach is based on the rigorous mathematical foundations of Morse theory and can be applied to a more general class of applications.

The association of E. coli and soil particles in overland flow.

The removal of E. coli from overland flow under saturation-excess runoff conditions was investigated in experimental field plots that were 1 m wide and 5 m long. Variation in the attenuation of bacteria and distance transported was quantified under contrasting flow conditions. In addition, the impact of soil tillage upon microbial attenuation was examined by comparing results derived from grassed plots (intact) with those subject to tillage with the soil left bare (cultivated). For intact plots subjected to a flow of 2 L/min, 27% of the E. coli in the flow was removed after 5 m with removal following a logarithmic function with respect to distance. For the higher flow rates of 6 L/min and 20 L/min, no attenuation trend was observed over this distance. E. coli removal during flow across the cultivated plots was significantly greater compared to the intact plots. This was attributed to a greater infiltration rate in the cultivated plots (due to the tillage) which promoted a greater volume of flow to pass through the soil matrix, providing the opportunity for filtration and adsorption of microbes. Logarithmic trends with respect to distance were observed for all flow rates tested on the cultivated plots (2, 6 and 20 L/min). Total removal after 5 m at a flow rate of 2 L/min was 41% and again removal efficiency decreased as the flow rate increased. Analysis of the transported state of the E. coli revealed that the bacteria were being transported predominantly in particles less than 20 microm in diameter and were not attached to large (dense) soil particles. The limited removal (< 50%) of bacteria from overland flow under saturation-excess runoff conditions in these experiments appeared, therefore, to be primarily due to a lack of settling or deposition. Instead, most bacteria remained entrained within the overland flow down the length of the plots.

Numbers and transported state of Escherichia coli in runoff direct from fresh cowpats under simulated rainfall.

AIMS: To investigate the number of Escherichia coli in runoff derived directly from fresh cowpats and to determine if the E. coli are attached to dense particles, in flocs or as individual cells. METHODS AND RESULTS: Three cowpats were collected monthly from the same farm for 13 months and the number of E. coli in them estimated. A rainfall simulator was used to generate runoff from the individual cowpats, which was fractioned to determine the transported state of any E. coli present. The number of E. coli in the cowpat runoff was highly variable and was strongly correlated with the number of E. coli in the cowpat. Only a small percentage (approx. 8%) of the E. coli in runoff were attached to dense (>1.3 g ml(-1)) particles and there was no evidence of flocculation of the cells. CONCLUSIONS: Escherichia coli in runoff from cowpats are transported predominantly as individual cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Mitigation strategies to reduce the number of faecal bacteria in overland flow from agricultural land need to be designed to trap single bacterial cells.

Survival of Campylobacter jejuni in water: effect of grazing by the freshwater crustacean Daphnia carinata (Cladocera).

Environmental studies of the human-pathogenic bacterium Campylobacter jejuni have focused on linking distributions with potential sources. However, in aquatic ecosystems, the abundance of C. jejuni may also be regulated by predation. We examine the potential for grazing by the freshwater planktonic crustacean Daphnia carinata to reduce the survival of C. jejuni. We use a system for measuring grazing and clearance rates of D. carinata on bacteria and demonstrate that D. carinata can graze C. jejuni cells at a rate of 7% individual(-1) h(-1) under simulated natural conditions in the presence of an algal food source. We show that passage of C. jejuni through the Daphnia gut and incorporation into fecal material effectively reduces survival of C. jejuni. This is the first evidence to suggest that grazing by planktonic organisms can reduce the abundance of C. jejuni in natural waters. Biomanipulation of planktonic food webs to enhance Daphnia densities offers potential for reducing microbial pathogen densities in drinking water reservoirs and recreational water bodies, thereby reducing the risk of contracting water-borne disease.

Evaluation of the effectiveness of a commercially available defined substrate medium and enumeration system for measuring Escherichia coli numbers in faeces and soil samples.

AIMS: To determine if a commercially available defined substrate medium and enumeration system could be utilized as an effective and accurate means of enumerating Escherichia coli in environmental samples containing faeces and soil. METHODS AND RESULTS: The samples tested were either inoculated with laboratory grown E. coli or natural E. coli populations in cow faeces. The number of E. coli recovered from faeces and soil samples using the defined substrate medium and enumeration system and a miniaturized MPN method (using traditional media) was compared by analysing the difference between the two methods in relation to the mean. For four of five groups of samples analysed there was no significant difference in the number of E. coli recovered by the two methods (P > 0.05). In one batch the difference was 0.30 log, which while being statistically significant (P < 0.01) was not considered to be biologically significant. CONCLUSION: The commercially available enumeration system was significantly more precise than the miniaturized MPN method (P < 0.001). SIGNIFICANCE AND IMPACT OF THE STUDY: We conclude that the commercially available defined substrate medium and enumeration system is a suitable method for the measurement of E. coli numbers in faeces and soil samples and should provide advantages of increased precision and a reduction in laboratory analysis time.