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J W Wimpenny

Publications and source records attributed to J W Wimpenny.

At least 19 recordsLinked to original sources

Individual-based modelling of biofilms.

Understanding the emergence of the complex organization of biofilms from the interactions of its parts, individual cells and their environment, is the aim of the individual-based modelling (IbM) approach. This IbM is version 2 of BacSim, a model of Escherichia coli colony growth, which was developed into a two-dimensional multi-substrate, multi-species model of nitrifying biofilms. It was compared with the established biomass-based model (BbM) of Picioreanu and others. Both models assume that biofilm growth is due to the processes of diffusion, reaction and growth (including biomass growth, division and spreading). In the IbM, each bacterium was a spherical cell in continuous space and had variable growth parameters. Spreading of biomass occurred by shoving of cells to minimize overlap between cells. In the BbM, biomass was distributed in a discrete grid and each species had uniform growth parameters. Spreading of biomass occurred by cellular automata rules. In the IbM, the effect of random variation of growth parameters of individual bacteria was negligible in contrast to the E. coli colony model, because the heterogeneity of substrate concentrations in the biofilm was more important. The growth of a single cell into a clone, and therefore also the growth of the less abundant species, depended on the randomly chosen site of attachment, owing to the heterogeneity of substrate concentrations in the biofilm. The IbM agreed with the BbM regarding the overall growth of the biofilm, due to the same diffusion-reaction processes. However, the biofilm shape was different due to the different biomass spreading mechanisms. The IbM biofilm was more confluent and rounded due to the steady, deterministic and directionally unconstrained spreading of the bacteria. Since the biofilm shape is influenced by the spreading mechanism, it is partially independent of growth, which is driven by diffusion-reaction. Chance in initial attachment events modifies the biofilm shape and the growth of single cells because of the high heterogeneity of substrate concentrations in the biofilm, which again results from the interaction of diffusion-reaction with spreading. This stresses the primary importance of spreading and chance in addition to diffusion-reaction in the emergence of the complexity of the biofilm community.

Algorithms↗

Effect of EPS on biofilm structure and function as revealed by an individual-based model of biofilm growth.

We have simulated a nitrifying biofilm with one ammonia and one nitrite oxidising species in order to elucidate the effect of various extracellular polymeric substance (EPS) production scenarios on biofilm structure and function. The individual-based model (IbM) BacSim simulates diffusion of all substrates on a two-dimensional lattice. Each bacterium is individually simulated as a sphere of given size in a continuous, three-dimensional space. EPS production kinetics was described by a growth rate dependent and an independent term (Leudeking-Piret equation). The structure of the biofilm was dramatically influenced by EPS production or capsule formation. EPS production decreased growth of producers and stimulated growth of non-producers because of the energy cost involved. For the same reason, EPS accumulation can fall as its rate of production increases. The patchiness and roughness of the biofilm decreased and the porosity increased due to EPS production. EPS density was maximal in the middle of the vertical profile. Introduction of binding forces between like cells increased clustering.

Ammonia↗

Adenylates and adenylate-energy charge in submerged and planktonic cultures of Salmonella enteritidis and Salmonella typhimurium.

Adenine nucleotide values and adenylate energy charge (AEC) were measured during the growth of Salmonella enteritidis and Salmonella typhimurium as submerged colonies in agarose gel and gelatin gel, and as planktonic cells in broth. Growth in all three systems showed similar trends with a ten-fold decrease in total adenylate pool during exponential growth, before attaining a fairly stable value throughout stationary phase. AEC values were generally low, (approximately 0.66), but did rise slightly during stationary phase. The large proportion of dead cells during early exponential phase may have contributed to the adenosine diphosphate and adenosine monophosphate pools, through cell lysis or excretion, and it is suggested that this was likely to account for the low values of AEC. In agarose and gelatin gelled cultures the percentage of adenosine triphosphate (ATP) in relation to the total adenylates showed random fluctuations. This was contrary to the broth culture where percentage ATP was highest after 12 h and the data formed a smooth curve. These data demonstrated that considerable physiological heterogeneity exists within a colony of bacteria growing in a gel matrix and by analogy a food material also, and that AEC is a poor indicator of cell viability in such systems.

Adenosine Triphosphate↗

Synergist effect of sucrose fatty acid esters on nisin inhibition of gram-positive bacteria.

Nisin in combination with the sucrose fatty acid esters, sucrose palmitate (P-1570 and P-1670) or sucrose stearate (S-1570 and S-1670) was tested against a range of Gram-negative and Gram-positive bacteria. Initial liquid culture investigation showed that the sugar ester P-1670 resulted in a synergist enhancement of the bacteriostatic activity of nisin against Gram-positive bacteria and not Gram-negative bacteria. Some enhancement of the bactericidal activity of nisin against Listeria monocytogenes was also observed. This increased nisin inhibitory effect was confirmed on solid media using plates with gradients of pH and NaCl. Synergism was observed with all four sucrose fatty acid esters, which enhanced the antimicrobial activity of nisin against several strains of L. monocytogenes, Bacillus cereus (both cells and spores), Lactobacillus plantarum and Staphylococcus aureus. The combination of nisin and the sucrose fatty acid esters showed no inhibition of Gram-negative bacteria (Salmonella enteritidis, Salm. typhimurium and Pseudomonas aeruginosa).

Anti-Bacterial Agents↗

The effects of agar concentration on the growth and morphology of submerged colonies of motile and non-motile bacteria.

The growth and morphology of submerged bacterial colonies was investigated. Five separate colonial forms were recognized depending both on species and on agar concentration. These were (i) branched, dendritic structures seen only with Bacillus cereus; (ii) lenticular colonies for all other species at high agar concentrations; (iii) small lobed to spherical colonies for non-motile organisms at low agar concentrations; (iv) and (v) large diffuse spherical colonies which can be further subdivided into 'snowball' or 'wispy' types for motile bacteria growing at agar concentrations below about 0.65% w/v. Viable count determinations suggested that agar concentration had little effect in the early stages of growth but that motile cells at low agar concentrations achieved higher cell numbers than did those in concentrations greater than 0.65% w/v. Transmission electron microscopy indicated that bacteria in lenticular colonies were tightly packed within lens-shaped splits in the agar whilst at low agar concentrations motile cells were well separated and appeared to move through the agar matrix.

Agar↗

The validity of models.

The ubiquity of biofilm and its classification as a microbial aggregate is discussed. Investigations into any microbial ecological problem operate at four levels: (i) in situ investigations, (ii) the use of microcosms, (iii) experimental model systems, and (iv) mathematical models. Each of these is defined and their use in biofilm research illustrated. It is concluded that all these approaches are valid and that scientific research in general and biofilm research in particular must profit by the use widely different methods if a complete understanding of a system is to be achieved.

Biofilms↗

The effects of growth dynamics upon pH gradient formation within and around subsurface colonies of Salmonella typhimurium.

pH measurements made in and around submerged colonies of Salmonella typhimurium grown within a model gelatin gel system using pH-sensitive micro- and macroelectrodes indicated some pH heterogeneity occurring in and around the bacterial colony. Inoculation density, initial pH and glucose concentration were all found to influence colony diameter and metabolism of Salmonella colonies. Colony growth in the presence of glucose, at pH 7.0 with an inoculation density of 1 cell ml-1 led to a pH fall of 1-2 pH units after 2 d. At pH 5.0, with glucose, colony growth rates were much slower than at pH 7.0, and the pH change varied by less than one pH unit often becoming alkaline. In the absence of glucose, only small pH changes were observed within the medium, although growth rates were similar to those in glucose-containing media. At the higher inoculation density (ca 1000 cells ml-1), isolated pH changes were not observed. Morphological changes, such as the production of annular rings, were noted in stationary phase colonies as was alkali production in colonies. These results are discussed in relation to observations with surface colonies.

Culture Media↗

Competition between Salmonella and Pseudomonas species growing in and on agar, as affected by pH, sodium chloride concentration and temperature.

pH/sodium chloride (NaCl) gradient plates were used to investigate competition between Pseudomonas and Salmonella species. At 30 degrees C and at particular NaCl/pH conditions the salmonellae inhibited growth of P. fluorescens and not P. putida. At 20 degrees C P. putida and not P. fluorescens inhibited the salmonellae. The growth of the pure and mixed strains in agar plates with the pH/NaCl conditions was compared by viable counts. Competition in pour plates at 30 degrees C was confirmed. At 20 degrees C, sub-surface growth of the salmonellae inhibited the P. putida. With surface growth this did not occur; the salmonellae was slightly inhibited by the P. putida.

Agar↗

The effect of chlorhexidine on defined, mixed culture oral biofilms grown in a novel model system.

In order to develop an improved method to evaluate antimicrobial agents for use in clinical dentistry, a constant-depth film fermenter (CDFF) has been used to generate biofilms of fixed depth comprising nine species of bacteria commonly found in dental plaque in health and disease. These bacteria were grown together initially in a conventional chemostat which was used to inoculate the CDFF over an 8 h period. Medium was then supplied directly to the CDFF and biofilms allowed to develop. The biofilms were then challenged with eight short pulses of two concentrations of chlorhexidine (0.0125 and 0.125% w/v). The lower concentration had a limited effect on the composition of the biofilms while a differential and substantial inhibition was obtained with a higher concentration. Actinomyces naeslundii was lost from the biofilm, and the viable counts of streptococci, Fusobacterium nucleatum and Porphyromonas gingivalis were inhibited by over three orders of magnitude by 0.125% chlorhexidine, whereas Veillonella dispar was only transiently affected. The findings were consistent with those from clinical studies of dental plaque, suggesting that this model would have a predictive value when evaluating novel antiplaque or antimicrobial inhibitors.

Anti-Infective Agents, Local↗

Investigation of the effect of combined variations in temperature, pH, and NaCl concentration on nisin inhibition of Listeria monocytogenes and Staphylococcus aureus.

Gradient plates were used to investigate the effects of varying temperature, pH, and sodium chloride (NaCl) concentration on nisin inhibition of Staphylococcus aureus and Listeria monocytogenes, Nisin was incorporated into the plates of 0, 50, 100, 250, and 500 IU ml -1. Gradients of pH (3.7 to 7.92) at right angles to NaCl concentration (2.1 to 7% [wt/vol]) were used for the plates, which were incubated at 20, 25, 30 and 35 degrees C. Growth on the plates were recorded by eye and by image analysis. The presence of viable but nongrowing cells was revealed by transfer to nongradient plates. Lower temperatures and greater NaCl concentrations increased the nisin inhibition of S. aureus synergistically. Increasing the NaCl concentration potentiated the nisin action against L. monocytogenes; the effect of temperature difference was not so apparent. Between pH 7.92 and ca. pH 5, a fall pH appeared to increase nisin's effectiveness against both organisms. At more acid pH values (ca. pH 4.5 to 5), the organisms showed resistance to both nisin and NaCl at 20 and 25 degrees C. Similar results were obtained with one-dimensional liquid cultures.

Anti-Bacterial Agents↗

The use of two-dimensional gradient plates to investigate the range of conditions under which conjugal plasmid transfer occurs.

Gel-stabilized two-dimensional gradient plates were used to study the effects of pH, salt concentration and temperature on the conjugal transfer of plasmid RP4 between strains of Escherichia coli and Pseudomonas putida. The combinations of pH and salt concentration that permitted conjugation were mapped as a two-dimensional growth area occupied by transconjugants following conjugation. This conjugation domain was less extensive than the areas that supported growth of the parental strains, and showed evidence for the interactive effects of pH and salt concentration in determination of conditions that permitted conjugation. The size and shape of the conjugation domain was influenced by time, temperature, the identities of the donor and recipient bacteria, and the combination of donor and recipient bacteria.

Conjugation, Genetic↗

Effect of three preservatives on the growth of Bacillus cereus, Vero cytotoxigenic Escherichia coli and Staphylococcus aureus, on plates with gradients of pH and sodium chloride concentration.

The effect of temperature, pH, sodium chloride concentration and a preservative (sodium benzoate, sodium nitrite or potassium sorbate) on the growth of three foodborne bacterial pathogens (Bacillus cereus, Vero cytotoxigenic Escherichia coli and Staphylococcus aureus) was studied using gradient gel plates. Growth, expressed in optical density units, was recorded using image analysis techniques, and was expressed as three-dimensional grids. These gave a visual indication of the effects of any three of the environmental factors on bacterial proliferation. Sorbate was completely effective against E. coli at all temperature/pH/NaCl combinations, and was the most effective preservative tested against B. cereus. Increase in the acidity and/or the NaCl concentration improved the effect of all the preservatives, except nitrite when used against St. aureus. Nitrite was the least effective preservative, particularly against St. aureus. At < 25 degrees C, sorbate was more effective than benzoate against St. aureus when used with higher concentrations of NaCl. At 35 degrees C benzoate was the most effective preservative against St. aureus, especially when used at pH < 6.

Bacillus cereus↗

Method for Investigation of Competition between Bacteria as a Function of Three Environmental Factors Varied Simultaneously.

Competition between microorganisms as affected by temperature, pH, and the sodium chloride (NaCl) concentration was investigated by selective replication from gradient plates. Salmonella typhimurium was inhibited by Pseudomonas putida at 20 and 23 degrees C but not 30 and 35 degrees C. P. putida no longer grew at the extremes of pH and NaCl concentration, particularly at 30 and 35 degrees C.

Journal Article↗

A novel directly coupled gradostat.

The original bidirectional compound chemostat (gradostat) described by Lovitt and Wimpenny has been simplified by making a more compact apparatus in which chemical gradients are established by diffusion between adjacent culture chambers. The experimental model (diffusion coupled (DC) gradostat) consisted of five chambers whose contents could be agitated by turbines rotating in the horizontal plane on a common shaft. Two biological experiments were designed to reveal the value of the DC gradostat. A methylotroph (Methylophilus methylotrophus) grown in a methanol gradient showed expected changes in cell viability as a function of position in the five vessel array. Cells of two species of photosynthetic bacteria (Rhodobacter capsulata and Rhodopseudomonas marina/agilis) with different salt sensitivities could be mixed and subsequently separated by the DC gradostat operating with a NaCl gradient of 0-3% w/v.

Bacteria↗

Measurements of the distribution of adenylate concentrations and adenylate energy charge across Pseudomonas aeruginosa biofilms.

Adenine nucleotide pools and adenylate energy charge distributions were determined by using a laboratory-generated quasi-steady-state Pseudomonas aeruginosa biofilm. The method used involved freezing and sectioning of the intact biofilm, followed by extraction and assay of the adenylates in the sectioned material. Results indicated an increase in adenylate energy charge of about 0.2 units from the bottom to the surface of the biofilm. However, energy charge values were generally low throughout the biofilm, reaching a maximum of only 0.6 units. Of the adenylates measured, AMP was the predominant nucleotide, especially in the deeper parts of the biofilm profile.

Adenosine Diphosphate↗

Effects of salt concentration on bacterial growth on plates with gradients of pH and temperature.

A temperature gradient device has been used to produce pH-temperature gradient plates. The response of Salmonella typhimurium and Bacillus cereus to continuous gradients of pH and temperature has been mapped. A third variable, sodium chloride, was investigated by incorporating it at different concentrations in a series of gradient plates. Both organisms exhibited an optimum temperature and pH range for maximum salt tolerance.

Bacillus cereus↗