In vitro inhibition of Clostridium difficile by commercial probiotics: A microcalorimetric study

The aim of the study was to investigate the influence of some commercial probiotics on the growth of Clostridium difficile using isothermal microcalorimetry, a technique which can monitor the real time growth of bacteria. Commercial probiotic strains and products, Lactobacillus acidophilus LA-5(®), Bifidobacterium lactis BB-12(®), Probio 7(®) and Symprove™ were co-cultured with C. difficile in Brain Heart Infusion (BHI) broth supplemented with 0.1% (w/v) l-cysteine hydrochloride and 0.1% (w/v) sodium taurocholate and monitored in the microcalorimeter. Pseudomonas aeruginosa NCIMB 8628 was also co-cultured with C. difficile and studied. The results indicated inhibition of C. difficile by the probiotics. The inhibition of C. difficile was shown to be pH-dependent using neutralized and unmodified cell free supernatant (CFS) produced by the probiotic strains. However, concentrated CFS of the probiotics also inhibited the intestinal pathogen in a non pH-dependent manner, likely due to specific antimicrobial substances produced. The results also indicated that C. difficile growth was greatly influenced by the presence of sodium taurocholate and by the pH of the medium. A medium pH of between 6.45 and 6.9 demonstrated maximum growth of the organism in the microcalorimeter

DETERMINATION OF THE THERMODYNAMIC PARAMETERS FOR TRANSFER OF ALKOXYPHENOLS FROM AQUEOUS-SOLUTION TO SDS MICELLES BY A TAYLOR-ARIS DIFFUSION TECHNIQUE

Using Taylor-Aris diffusion techniques, thermodynamic parameters have been measured for the transfer of a series of alkoxyphenols from water into SDS micelles. The results are compared with those for transfer to bulk organic solvents and to cells of Escherichia coli. The SDS micelles are found to be marginally more polar than n-octanol, but the results reveal significant differences between bulk solvents and the more ordered micellar pseudophase.87172741274

An in vitro test of the efficacy of an anti-biofilm wound dressing

Broad-spectrum antimicrobial agents, such as silver, are increasingly being formulated into medicated wound dressings in order to control colonization of wounds by opportunistic pathogens. Medicated wound dressings have been shown in-vitro to be effective against planktonic cultures, but in-vivo bacteria are likely to be present in biofilms, which makes their control and eradication more challenging. Recently, a functional wound dressing (AQUACEL(®) Ag+ Extra™ (AAg + E)) has been developed that in addition to silver contains two agents (ethylenediaminetetraacetic acid (EDTA) and benzethonium chloride (BC)) designed to disrupt biofilms. Here, the efficacy of AAg + E is demonstrated using a biofilm model developed in an isothermal microcalorimeter. The biofilm was seen to remain viable in the presence of unmedicated dressing, silver-containing dressing or silver nitrate solution. In the presence of AAg + E, however, the biofilm was eradicated. Control experiments showed that neither EDTA nor BC alone had a bactericidal effect, which means it is the synergistic action of EDTA and BC disrupting the biofilm with silver being bactericidal that leads to the product's efficacy

Development of a flow system for studying biofilm formation on medical devices with microcalorimetry

Isothermal microcalorimetry (IMC) is particularly suited to the study of microbiological samples in complex or heterogeneous environments because it does not require optical clarity of the sample and can detect metabolic activity from as few as 10(4) CFU/mL cells. While the use of IMC for studying planktonic cultures is well established, in the clinical environment bacteria are most likely to be present as biofilms. Biofilm prevention and eradication present a number of challenges to designers and users of medical devices and implants, since bacteria in biofilm colonies are usually more resistant to antimicrobial agents. Analytical tools that facilitate investigation of biofilm formation are therefore extremely useful. While it is possible to study pre-prepared biofilms in closed ampoules, better correlation with in vivo behaviour can be achieved using a system in which the bacterial suspension is flowing. Here, we discuss the potential of flow microcalorimetry for studying biofilms and report the development of a simple flow system that can be housed in a microcalorimeter. The use of the flow system is demonstrated with biofilms of Staphylococcus aureus

Rapid diagnosis of experimental meningitis by bacterial heat production in cerebrospinal fluid

BACKGROUND: Calorimetry is a nonspecific technique which allows direct measurement of heat generated by biological processes in the living cell. We evaluated the potential of calorimetry for rapid detection of bacterial growth in cerebrospinal fluid (CSF) in a rat model of bacterial meningitis. METHODS: Infant rats were infected on postnatal day 11 by direct intracisternal injection with either Streptococcus pneumoniae, Neisseria meningitidis or Listeria monocytogenes. Control animals were injected with sterile saline or heat-inactivated S. pneumoniae. CSF was obtained at 18 hours after infection for quantitative cultures and heat flow measurement. For calorimetry, 10 microl and 1 microl CSF were inoculated in calorimetry ampoules containing 3 ml trypticase soy broth (TSB). RESULTS: The mean bacterial titer (+/- SD) in CSF was 1.5 +/- 0.6 x 108 for S. pneumoniae, 1.3 +/- 0.3 x 106 for N. meningitidis and 3.5 +/- 2.2 x 104 for L. monocytogenes. Calorimetric detection time was defined as the time until heat flow signal exceeded 10 microW. Heat signal was detected in 10-microl CSF samples from all infected animals with a mean (+/- SD) detection time of 1.5 +/- 0.2 hours for S. pneumoniae, 3.9 +/- 0.7 hours for N. meningitidis and 9.1 +/- 0.5 hours for L. monocytogenes. CSF samples from non-infected animals generated no increasing heat flow (<10 microW). The total heat was the highest in S. pneumoniae ranging from 6.7 to 7.5 Joules, followed by L. monocytogenes (5.6 to 6.1 Joules) and N. meningitidis (3.5 to 4.4 Joules). The lowest detectable bacterial titer by calorimetry was 2 cfu for S. pneumoniae, 4 cfu for N. meningitidis and 7 cfu for L. monocytogenes. CONCLUSION: By means of calorimetry, detection times of <4 hours for S. pneumoniae and N. meningitidis and <10 hours for Listeria monocytogenes using as little as 10 microl CSF were achieved. Calorimetry is a new diagnostic method allowing rapid and accurate diagnosis of bacterial meningitis from a small volume of CSF

Observation with microcalorimetry: Behaviour of P. aeruginosa in mixed cultures with S. aureus and E. coli

In this work, isothermal microcalorimetry (IMC) was used to study the behaviour of mixed cultures of Pseudomonas aeruginosa with Staphylococcus aureus and Escherichia coli. The species were firstly studied as pure culture at 10 6 CFU/mL densities in nutrient broth (NB). The species were then introduced into NB to give an equivalent inoculum of 10 6 CFU/mL of each. Additionally, the inoculum density of one species was maintained at 10 6 CFU/mL and the other varied between 10 3 –10 5 CFU/mL. The IMC power-time curves were characteristic for the species in the medium. The total heat output (Q t ), amplitude of first peak (P 1 ), time of registration of first peak (t 1 ), final metabolic/maximum peak (P fmax ) and time of registration of the final metabolic/maximum peak (t fmax ) were calculated from the power-time data. A mixed culture of P. aeruginosa and S. aureus at equal densities exhibited metabolism synonymous to P. aeruginosa alone. When the density of P. aeruginosa was decreased, S. aureus gradually recovered showing power-time profiles that demonstrated this. Mixed cultures of P. aeruginosa and E. coli at equal densities showed power-time profiles representative of both species

SOLUTION THERMODYNAMICS FOR ALKOXY PHENOLS IN ALCOHOL AND IN WATER-ALCOHOL SYSTEMS

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A THERMODYNAMIC ANALYSIS OF THE COLLANDER EQUATION AND ESTABLISHMENT OF A REFERENCE SOLVENT FOR USE IN DRUG PARTITIONING STUDIES

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