Comparative evaluation of agar dilution and broth microdilution by commercial and in-house plates for Bacteroides fragilis group: An economical and expeditious approach for resource-limited settings.

OBJECTIVE: To compare the performance of agar dilution and broth microdilution by commercial and in-house prepared plates for the Bacteroides fragilis group. The cost analysis was performed to demonstrate that in-house prepared BMD plates were a suitable alternative to agar dilution given the high cost and low feasibility of incorporating commercial BMD plates in routine, particularly in the tertiary care institutes of many low- and middle-income countries. METHODS: Thirty B. fragilis group isolates were tested against six antibiotics, frequently used as empirical therapy for anaerobic infections including metronidazole, clindamycin, imipenem, piperacillin-tazobactam, cefoxitin, and chloramphenicol. The running consumable expenditure for all methodologies was calculated. RESULTS: The results demonstrated essential and categorical agreement of >90% for all antibiotics except cefoxitin, which showed <90% categorical agreement. No major or very major errors were observed. We observed a high agreement and strong concordance for MIC values between both methods and inter-rate reliability of >0.9 by Cohen's kappa analysis, indicating almost perfect agreement between both methods using either of the plates. In contrast to agar dilution, a 20.5 fold cost reduction was seen in BMD using in-house plates and a 5.8 fold reduction using commercial plates to test a single isolate. However, when testing 30 isolates concurrently the cost significantly increased for commercial BMD plates by 8.4 folds, and only 1.03 fold cost reduction was seen with in-house BMD plates. CONCLUSION: BMD gives comparable results to agar dilution and can be considered a method of choice to test a small number of samples. The technique is an economical option when plates are standardized in-house and could be employed for susceptibility testing of the B. fragilis group.

Seaweed Polysaccharide Based Products and Materials: An Assessment on Their Production from a Sustainability Point of View.

Among the various natural polymers, polysaccharides are one of the oldest biopolymers present on the Earth. They play a very crucial role in the survival of both animals and plants. Due to the presence of hydroxyl functional groups in most of the polysaccharides, it is easy to prepare their chemical derivatives. Several polysaccharide derivatives are widely used in a number of industrial applications. The polysaccharides such as cellulose, starch, chitosan, etc., have several applications but due to some distinguished characteristic properties, seaweed polysaccharides are preferred in a number of applications. This review covers published literature on the seaweed polysaccharides, their origin, and extraction from seaweeds, application, and chemical modification. Derivatization of the polysaccharides to impart new functionalities by chemical modification such as esterification, amidation, amination, C-N bond formation, sulphation, acetylation, phosphorylation, and graft copolymerization is discussed. The suitability of extraction of seaweed polysaccharides such as agar, carrageenan, and alginate using ionic solvent systems from a sustainability point of view and future prospects for efficient extraction and functionalization of seaweed polysaccharides is also included in this review article.

Assessment of carbapenem-resistant Enterobacteriaceae-plate formula and quality control procedure.

AIMS: To assess a cost-effective in-house selective plate formula for actively screening carbapenem-resistant Enterobacteriaceae (CRE). METHODOLOGY AND RESULTS: The in-house formula included CHROMagarTM Orientation, meropenem, and ingredients present in the Mac-Conkey formula, such as bile salts and crystal violet (pH 6.9-7.2). American Type Culture Collection strains and 200 clinical strains were used to validate the plate formula. The CRE plates had a sensitivity of 97.4% and a specificity of 98.8% with ATCC andor clinical strains used in the quality control procedure. A point prevalence survey among the 18 inpatients at Viet-Tiep hospital ICU using fecal swabs plated at the in-house agar plate showed a CRE prevalence of 44.4%. CONCLUSION: The in-house plate had high sensitivity and specificity, particularly for Escherichia coli and the KESC group (Klebsiella spp., Enterobacter spp., Serratia marscescens, and Citrobacter spp.), and it may be widely applied as an alternative to other ready-to-use commercial plates. SIGNIFICANCE AND IMPACT OF THE STUDY: The formula developed in the present study may facilitate the early detection and isolation of CRE and decrease transmission, particularly in low- and middle-income countries with a high rate of CRE colonization and limited access to ready-to-use commercial plates.

Inoculation of Ensifer fredii strain LP2/20 immobilized in agar results in growth promotion and alteration of bacterial community structure of Chinese kale planted soil.

In our former research, we succeeded in using agar, alginate, and perlite as immobilization materials to maintain long-term survival of the inoculant, Ensifer fredii LP2/20, in a controlled glasshouse. Therefore the information on the establishment and activity of the inoculant to promote plant growth under field conditions, the effects of the inoculant on the soil microbial communities and specific microbial taxa, and the association between the inoculant and soil elements merit further studies. Here, we found that agar was the most suitable material that supported the establishment of the inoculant under field conditions. RNA-based analysis showed that E. fredii LP2/20 immobilized in agar was still metabolically active at day 50 after being introduced into soil. Inoculation of E. fredii LP2/20 immobilized in agar conferred the highest plant dry weight (up to 89.94%) and all plant elements including total N (9.55%), P (17.94%), K (68.42%), Ca (39.77%), Mg (30.76%), Fe (29.85%), and Zn (22.44%). Inoculation of E. fredii LP2/20 immobilized in agar increased soil chemicals including soil organic matter (99.02%), total N (272.48%), P (31.75%), K (52.74%), Fe (51.06%), and Zn (63.10%). High-throughput next-generation sequencing of bacterial 16S rRNA amplicons showed that the Proteobacteria, Acidobacteria, Bacteroidetes, and Firmicutes were dominant phyla in Chinese kale field soil. Inoculation of E. fredii LP2/20 significantly affected the soil bacterial community structure by decreasing total bacterial richness and diversity. The numbers of alpha- and gamma-Proteobacteria were significantly increased while the number of delta-Proteobacteria was significantly decreased due to E. fredii LP2/20 establishment. Soil total P, K, and Ca and soil pH were the important factors that shaped the soil bacterial community composition.

Comparison of four agar media for the enumeration of the Mycobacterium abscessus complex.

Background: Traditional culture of nontuberculous mycobacteria (NTMs) has involved egg-based formulations (Lowenstein-Jensen medium) or defined media (Middlebrook formulations), which have disadvantages of composition complexity, availability, and cost. This study quantitatively compared three non-selective, non-blood based basal agars with Columbia blood agar (CBA), to enumerate Mycobacterium abscessus complex organisms in pure culture. Methods: M. abscessus subsp. massiliense, M. abscessus subsp. bolletii, and M. abscessus subsp. abscessus were employed. Inocula of each of these were counted on three basal agar media, including (i) standard plate count agar (SPCA), (ii) tryptone soya agar (TSA), and (iii) Mueller-Hinton agar (MHA) and compared to counts on CBA. Results: All NTM isolates of all subspecies grew successfully on all four media examined. The growth was most profuse on SPCA, with a mean colony diameter of 3 mm, whereas the mean colony diameter on all other media was 1 mm. Statistically, there was no significant difference in counts when comparing CBA with SPCA or MHA (P > 0.05), whereas there was a statistically significant difference between CBA and TSA (P = 0.01). There was no statistically significant difference between SPCA and MHA (P = 0.53). Conclusion: This study indicates that SPCA and MHA are equally effective as CBA, when enumerating of M. abscessus complex organisms. Employment of TSA gave significantly lower counts than CBA (P = 0.01) and therefore should not be employed when enumerating these organisms. SPCA yielded the most profuse growth of all media examined. In addition to these advantages, given that SPCA does (i) not require blood as a medium constituent, (ii) is simple to reconstitute, (iii) is relatively cheap, and (iv) is widely available commercially, this study endorses employment of SPCA for the nonselective culture of M. abscessus complex organisms, including enumeration.

[Criterium pencil mines as an alternative to platinum rods (Inoclic) in achievement of the antibiogram in agar medium for countries with limited resources?] / Les mines de crayon pour critérium comme alternative aux tiges de platine (Inoclic) dans la réalisation de l'antibiogramme en milieu gélosé pour les pays à ressources limitées ?

The realization of the antibiotic susceptibility test in agar is the routine bacteriological examination for the determination and monitoring of bacterial susceptibility to antibiotics. In this study, we report the comparative results between pencil leads for criterium, as an alternative to platinum rods in the realization of the antibiotic susceptibility test. METHODOLOGY: Experimental study evaluating the comparability of the results between Criterium and Inoclic mines (by counting bacterial cells on agar after 5 successive dilutions of reason 10 from a bacterial suspension obtained after piercing through a colony; by measuring the inhibition diameters of 4 ATCC reference bacterial strains on an antibiogram in an agar medium) and evaluating the sterility of the criterium mines by culturing them on enriched broth (heart - brain type). RESULTS: 42 bacterial strains were used for bacterial cell counting. The results were of the same order of magnitude (107 CFU/mL) between Inoclic and criterium mines, for all strains and at all dilutions. The antibiotic susceptibility tests performed for the 4 reference strains by the Inoclics and criterium mines all complied (100%) with the expected limits for determining their sensitivity profile to the antibiotics tested. Compared to the bacterial growth inhibition diameters on antibiotic susceptibility tests, no intra-operator variability was observed, while significant inter-operator variability (both with Inoclic and 0.5 mm criterium mines) was observed with some strains and for inhibition diameters greater than 10 mm. The enriched broth cultures (BCC) and their subculture carried out on 10 criterium mines from 5 different batches were negative. CONCLUSION: Criterium mines seem to be a serious and less expensive alternative to Inoclic for the realization of antibiotic susceptibility testing in our resource-limited countries.

Performance of Low-Cost Agar from Gracilaria salicornia on Tissue Culture of Pleurotus HK-37.

Currently, the demand of Pleurotus HK-37 (oyster mushroom) in Tanzania is growing rapidly due to the increasing of awareness on its nutrition, health, and economic benefits. Despite the increasing demand, the availability of strains of Pleurotus HK-37 species is still a challenge due to high cost of tissue culture technology. The high cost of importing agar seems to be among the factors for this failure. This study aimed at investigating the performance of low-cost agar from local Gracilaria salicornia on tissue culture of Pleurotus HK-37. Local extracted agars with different gel strengths ranging between 100, 200, 300, 400, and 500 g/cm2 were used to make PDA media. The average mycelia growth rate (mm/day) ranged between 9.87 ± 1.44 and 14.9 ± 0.85 mm/day. Low-cost agar shows quite similar performance as that of standard agar on active growth of Pleurotus HK-37 mycelia. All PDA plates appeared white and feathery and showed to grow in a circular mode (radial extension). Mycelia growth on standard agar PDA took 5 days while on extracted local agar PDA took 5 to 7 days to fully colonize the plate at 27 ± 2°C. The present study shows that the production cost can be reduced by ∼35-78% by using local agar.

Development of a novel and economical agar-based non-adherent three-dimensional culture method for enrichment of cancer stem-like cells.

BACKGROUND: Non-adherent or ultra-low attachment three-dimensional (3D) culture, also called sphere formation assay, has been widely used to assess the malignant phenotype and stemness potential of transformed or cancer cells. This method is also popularly used to isolate the cancer stem-like cells (CSCs) or tumor-initiating cells based on their unique anchorage-independent growth or anoikis-resistant capacity. Different non-adhesive coating agents, such as poly-2-hydroxyethyl methacrylate (poly-HEMA) and synthetic hydrogels, have been used in this non-adherent 3D culture. However, preparation of non-adherent culture-ware is labor-intensive and technically demanding, and also costs of commercial non-adherent culture-ware prepared with various coating agents are relatively expensive and the culture-ware cannot be used repeatedly. METHODS: In this study, we developed a non-adherent 3D culture method based on agar coating for growing tumor spheres derived from various cancer cell lines and primary prostate cancer tissues under a non-adherent and serum-free condition. The tumor spheres generated by this 3D culture method were analyzed on their expression profiles of CSC-associated markers by reverse transcription quantitative polymerase chain reaction, presence and relative proportion of CSCs by fluorescence-activated cell sorting (CD133+/CD44+ cell sorting) and also a CSC-visualizing reporter system responsive to OCT4 and SOX2 (SORE6), and in vivo tumorigenicity. The repeated use of agar-coated plates for serial passages of tumor spheres was also evaluated. RESULTS: Our results validated that the multicellular tumor spheres generated by this culture method were enriched of CSCs, as evidenced by their enhanced expression profiles of CSC markers, presence of CD133+/CD44+ or SORE6+ cells, enhanced self-renewal capacity, and in vivo tumorigenicity, indicating its usefulness in isolation and enrichment of CSCs. The agar-coated plates could be used multiple times in serial passages of tumor spheres. CONCLUSIONS: The described agar-based 3D culture method offers several advantages as compared with other methods in isolation of CSCs, including its simplicity and low-cost and repeated use of agar-coated plates for continuous passages of CSC-enriched spheres.

The visual assessment of broth cultures for tissue bank samples.

The bioburden screening process of allograft musculoskeletal tissue samples received at the South Eastern Area Laboratory Services includes the routine use of solid agar and cooked meat (CM) broth media. CM has been routinely sub-cultured onto solid agar plates after aerobic incubation at 35 °C. This study will evaluate whether a visual assessment of CM can replace sub-culture by an in vitro inoculation and a prospective study. Eight challenge organisms were serially diluted and inoculated into CM. The average inoculum of 0.5-5.5 CFU produced visible turbidity of CM after 24-h incubation for 7 of the challenge organisms with one organism producing turbidity after 48-h incubation. The prospective study evaluated 222 CM of which 213 were visually clear and no-growth on sub-culture and 9 turbid CM which were culture positive. Broth cultures are an integral part of the bioburden screening process of allograft musculoskeletal tissue and swab samples and visual assessment of CM can replace sub-culture.

Cell-on-hydrogel platform made of agar and alginate for rapid, low-cost, multidimensional test of antimicrobial susceptibility.

Antimicrobial resistance (AMR) is a rapidly increasing threat to the effective treatment of infectious diseases worldwide. The two major remedies include: (1) using narrow-spectrum antibiotics based on rapid diagnosis; and (2) developing new antibiotics. A key part of both remedies is the antimicrobial susceptibility test (AST). However, the current standard ASTs that monitor colony formation are costly and time-consuming and the new strategies proposed are not yet practical to be implemented. Herein, we report a strategy to fabricate whole-hydrogel microfluidic chips using alginate-doped agar. This agar-based microfabrication makes it possible to prepare inexpensive hydrogel devices, and allows a seamless link between microfluidics and conventional agar-based cell culture. Different from common microfluidic systems, in our system the cells are cultured on top of the device, similar to normal agar plate culture; on the other hand, the microfluidic channels inside the hydrogel allow precise generation of linear gradient of drugs, thus giving a better performance than the conventional disk diffusion method. Cells in this system are not exposed to any shear flow, which allows the reliable tracking of individual cells and AST results to be obtained within 2-3 hours. Furthermore, our system could test the synergistic effect of drugs through two-dimensional gradient generation. Finally, the platform could be directly implemented to new drug discovery and other applications wherein a fast, cost-efficient method for studying the response of microorganisms upon drug administration is desirable.

Characterization of a dielectric phantom for high-field magnetic resonance imaging applications.

PURPOSE: In this work, a generic recipe for an inexpensive and nontoxic phantom was developed within a range of biologically relevant dielectric properties from 150 MHz to 4.5 GHz. METHODS: The recipe includes deionized water as the solvent, NaCl to primarily control conductivity, sucrose to primarily control permittivity, agar-agar to gel the solution and reduce heat diffusivity, and benzoic acid to preserve the gel. Two hundred and seventeen samples were prepared to cover the feasible range of NaCl and sucrose concentrations. Their dielectric properties were measured using a commercial dielectric probe and were fitted to a 3D polynomial to generate a recipe describing the properties as a function of NaCl concentration, sucrose concentration, and frequency. RESULTS: Results indicated that the intuitive linear and independent relationships between NaCl and conductivity and between sucrose and permittivity are not valid. A generic polynomial recipe was developed to characterize the complex relationship between the solutes and the resulting dielectric values and has been made publicly available as a web application. In representative mixtures developed to mimic brain and muscle tissue, less than 2% difference was observed between the predicted and measured conductivity and permittivity values. CONCLUSIONS: It is expected that the recipe will be useful for generating dielectric phantoms for general magnetic resonance imaging (MRI) coil development at high magnetic field strength, including coil safety evaluation as well as pulse sequence evaluation (including B1(+) mapping, B1(+) shimming, and selective excitation pulse design), and other non-MRI applications which require biologically equivalent dielectric properties.

Cost-effective nanoporous Agar-Agar polymer/Nickel powder composite particle for effective bio-products adsorption by expanded bed chromatography.

In the present work a novel kind of dense nanoporous composite matrix for expanded bed application has been successfully first prepared with Nickel powder as a densifier and was covered with Agar-Agar layer as a skeleton, through the method of water-in-oil emulsification. Agar-Agar is a porous and inexpensive polymer. In order to fabricate cost-effective adsorbent with favorable qualities Agar-Agar polymer was used. Thereafter, the customized composite particle was modified by pseudo-affinity dye-ligand, Reactive Blue 4 (RB4), aimed at preparing a pseudo-affinity adsorbent (RB4-Agar-Ni) for bioprodut adsorption from aqueous solution. Bovine Serum Albumin (BSA) was selected as a model protein to investigate the adsorption behavior in batchwise and expanded bed chromatography, and the obtained results were evaluated with that of Streamline™ (Amersham-Pharmacia Biotech, Sweden). Spherical appearance and porous structure of composite particles were observed by the optical microscope (OM) and scanning electronic microscope (SEM). The results suggested that the matrices followed the logarithmic normal size distribution with the range of 65-300 µm and average diameter of 126.81-151.47 µm, proper wet density of 1.64-2.78 g/ml, water content of 62.74-34%, porosity of 98-90% and pore size of about 38-130 nm. For better comprehension of the impact of solid phase properties on the performance of the expanded bed, the expansion and hydrodynamic properties of a composite matrix with a series of densities was evaluated and estimated by the retention time distribution method (RTD) in an expanded bed and was compared with that of other matrices. According to obtained results the expansion factors under the same fluid velocity decreased by increasing the matrix density. Moreover, the axial dispersion coefficient (Dax) is the most appropriate parameter for evaluating the stability of expanded bed, on various operating conditions, such as different flow velocity, bed expansion degree, viscosity of the liquid phase and the density of adsorbent. It was observed that the application of matrix with high density was proper for high operation, fluid velocity, since the addition of densifier improves the rigidity of the matrix. Three momentous factors, pH, ionic strength and initial concentration of BSA were analyzed. The best results showed that the adsorption equilibrium isotherms seems to follow a typical Langmuir isotherm and also the maximum adsorption capacity (qm) of BSA on RB4-Agar-Ni (64.01 mg/ml adsorbent) was higher than that on RB4-Streamline commercial adsorbent (about 54 mg.ml adsorbent). Additionally kinetic adsorption processes were characterized by the pseudo-first-order and pseudo-second-order kinetics equations. The experimental data followed the pseudo-first-order kinetic equation. Also the breakthrough curves were investigated. It was found that dynamic binding capacity (DBC) decreased with increasing the flow rate and the values of DBC decreased from 21.08 to 11.15 mg/ml adsorbent when the density of composite beads increased from 1.64 to 2.78 g/ml. All results indicate that the prepared composite is promising for efficient bioproduct adsorption with good hydrodynamic characteristics, high stability and it is suitable for expanded bed usage as a cost-effective adsorbent.

Assessment of the suitability of mannitol salt agar for growing bovine-associated coagulase-negative staphylococci and its use under field conditions.

This study aimed at testing the applicability of mannitol salt agar (MSA), a medium generally used in human medicine for differentiating Staphylococcus aureus from coagulase-negative staphylococci (CNS), for culturing bovine-associated CNS species. All test isolates from a comprehensive collection of well-identified CNS species, including both reference strains and field isolates, were able to grow. Subsequently, bulk milk samples and teat apex swabs were used to examine the capability of MSA for yielding CNS under field conditions. Sixty-nine and 47 phenotypically different colonies were retrieved from bulk milk and teat apices, respectively. The majority of isolates from teat apices were staphylococci, whereas in bulk milk, staphylococci formed a minority. After 24h of growth, recovery of separate colonies of CNS was much more convenient on MSA compared to a non-selective blood agar. The results of this study indicate that MSA is a suitable medium for both growth and recovery of bovine-associated CNS.

Investigating the effectiveness of economically sustainable carrier material complexes for marine oil remediation.

The application of bioremediation to marine oil spills is limited due to dilution of either nutrients or hydrocarbonoclastic organisms. This study investigated the effectiveness of three unique natural carrier materials (mussel shells, coir peat and mussel shell/agar complex) which allowed nutrients, hydrocarbonoclastic organisms and oil to be in contact, facilitating remediation. TPH analysis after 30 d showed that mussel shells exhibited the greatest capacity to degrade oil with a 55% reduction (123.3 mg l(-1) from 276 mg l(-1)) followed by mussel shell/agar complex (49%) and coir peat (36%). Both the mussel shells and mussel shell/agar complex carriers were significantly different to the control (P=0.008 and P=0.002, respectively). DGGE based cluster analysis of the seawater microbial community showed groupings based on time rather than carriers. This study demonstrated that inexpensive, accessible waste materials used as carriers of hydrocarbonoclastic bacteria led to significant degradation of hydrocarbon contaminants in seawater.

Back to the kitchen: food-grade agar is a low-cost alternative to bacteriological agar.

Food-grade agar can be used as a low-cost substitute for bacteriological agar in the preparation of solid microbial media. No difference was observed in the colony morphology, growth rate, or viability of bacteria grown on solid media prepared using food-grade agar as compared with using bacteriological-grade agar. This simple tip can reduce the cost of the most common solid media by 80% or more.

Bi-exponential magnetic resonance signal model for partial volume computation.

Accurate quantification of small structures in magnetic resonance (MR) images is often limited by partial volume (PV) effects which arise when more than one tissue type is present in a voxel. PV may be critical when dealing with changes in brain anatomy as the considered structures such as gray matter (GM) are of similar size as the MR spatial resolution. To overcome the limitations imposed by PV effects and achieve subvoxel accuracy different methods have been proposed. Here, we describe a method to compute PV by modeling the MR signal with a biexponential linear combination representing the contribution of at most two tissues in each voxel. In a first step, we estimated the parameters (T1, T2 and proton density) per tissue. Then, based on the bi-exponential formulation one can retrieve fractional contents by solving a linear system of two equations with two unknowns, namely tissue magnetizations. Preliminary tests were conducted on images acquired on a specially designed physical phantom for the study of PV effects. Further, the model was tested on BrainWeb simulated brain images to estimate GM and white matter (WM) PV effects. Root mean squared error was computed between the BrainWeb ground truth and the obtained GM and WM PV maps. The proposed method outperformed traditionally used methods by 33% and 34% in GM and WM, respectively.

[Evaluation of chromogenic medium Uriselect4 in urine culture]. / Évaluation du milieu chromogène Uriselect4 au cours des urocultures.

We evaluated the performance and the cost of chromogenic medium Uriselect4 agar with regard to the standard medium for the detection and identification of urinary tract pathogens. A total of 503 clinical urine specimens containing leucocytes greater or equal to 104/mL were analysed prospectively, in parallel by two different persons on blood agar (GS) and Uriselect4 according to the manufacturers' instructions. Of the 503 urine specimens tested, 210 gave a positive culture on Uriselect4 versus 181 on GS. The majority of bacterial species grew on both media; enterobacteria grew on Uriselect4 better than GS. The identification of Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), KES group and Enterococcus faecalis (E. faecalis) did not require the use of galleries Api and has a gain of 24  h. Positive pure cultures on Uriselect4 corresponding to negative cultures of GS were noted in 17 ases. Conversely, in seven cases a positive pure culture on GS was noted while the corresponding Uriselect4 cultures were negative. The cost of identification on GS (including the cost of galleries Api), was about two times higher than Uriselect4. Uriselect4 medium isolates the most frequent urinary tract pathogens and identify them so almost immediately, with a lower cost.

Occurrence of Escherichia coli O157, O111 and O26 in raw ewe's milk and performance of two enrichment broths and two plating media used for its assessment.

The occurrence of Escherichia coli O157, O111 and O26 in 159 raw ewe's milk samples was examined. Sample-aliquots were incubated simultaneously in TSB added with yeast extract (YETSB) and mTSB with novobiocin (N-mTSB). Serogroup-specific immunomagnetic separation (IMS) was then used and IMS beads were plated in a cefixime tellurite (CT)-containing media (CT-SMAC, CT-SBMAC and CT-RMAC for E. coli O157, O111 and O26, respectively) and E. coli O157:H7 chromogenic ID agar. A sweep of confluent growth from each medium was examined for the presence of E. coli O157 and O111 using PCR, and for E. coli O26 using a latex agglutination test. Enumeration of E. coli O157 and O111 was performed in the samples tested positive for the correspondent serogroup using the most probable number (MPN) method combined with PCR. Percentage occurrences of E. coli O157, O111 and O26 were 18.2, 8.2 and 5.7, respectively. Mean E. coli O157 and O111 levels were 0.22 and <0.04 MPN/mL, respectively. Enrichment in YETSB resulted in higher detection rates of E. coli O157 and O26 than in N-mTSB. When YETSB was used as enrichment broth and for these last two serogroups, the analysis of the confluent growth from the CT-media gave more positive results than that from E. coli O157:H7-ID medium.

Modelling conjugation with stochastic differential equations.

Conjugation is an important mechanism involved in the transfer of resistance between bacteria. In this article a stochastic differential equation based model consisting of a continuous time state equation and a discrete time measurement equation is introduced to model growth and conjugation of two Enterococcus faecium strains in a rich exhaustible media. The model contains a new expression for a substrate dependent conjugation rate. A maximum likelihood based method is used to estimate the model parameters. Different models including different noise structure for the system and observations are compared using a likelihood-ratio test and Akaike's information criterion. Experiments indicating conjugation on the agar plates selecting for transconjugants motivates the introduction of an extended model, for which conjugation on the agar plate is described in the measurement equation. This model is compared to the model without plate conjugation. The modelling approach described in this article can be applied generally when modelling dynamical systems.

Selective adsorbent for the removal of immunoglobulin E in bronchial asthma.

Agar-gel resin with tryptophan as ligand was synthesized as an adsorbent for the removal of Immunoglobulin E (IgE) in bronchial asthma. Factors affecting the adsorption properties of the adsorbent, such as the different kind of amino acid linked to the carrier, NaOH concentration and amount of epichlorohydrin used in activation, were studied in detail. The effect of spacer on the adsorption capacity was also investigated. Results showed that the agar-tryptophan adsorbent with a spacer could enhance the adsorption capacity to 70.73% (1309.403IU/g agar-gel), which was higher than that without spacers (53%). The selectivity and hemocompatibility of the adsorbent were also examined, which showed good selectivity for IgE and satisfactory blood compatibility.