Epidemiological Characterization of Group B Streptococcus Infections in Alberta, Canada: An Update from 2014 to 2020.

Group B Streptococcus (GBS) is a leading cause of invasive neonatal disease. Epidemiological surveillance of GBS is important to determine cumulative incidence, antimicrobial resistance rates, and maternal and neonatal disease prevention. In this study, we present an update on GBS epidemiology in Alberta, Canada, from 2014 to 2020. Over the 7-year period, 1,556 GBS isolates were submitted to the Alberta Public Health Laboratory for capsular polysaccharide (CPS) typing and antimicrobial susceptibility testing. We analyzed the distribution of CPS types in Alberta and found CPS types III (23.6%), Ia (16.0%), Ib (14.8%), II (13.3%), V (12.7%), IV (12.5%), and VI (2.38%) to be the most prevalent. Less than 1% each of CPS types VII, VIII, and IX were identified. In agreement with historical data, the presence of CPS type IV continued to rise across Alberta, particularly in cases of adult infection, where a 2-fold increase was observed. Cumulative incidences of GBS cases per 100,000 population and late-onset disease per 1,000 live births increased from 4.43 to 5.36 and 0.38 to 0.41, respectively, from 2014 to 2020. However, the incidence of early-onset disease decreased during the 7-year period from 0.2 to 0.07, suggestive of successful intrapartum chemoprophylaxis treatment programs. All GBS isolates were susceptible to penicillin and vancomycin. However, nonsusceptibility to erythromycin increased significantly, from 36.85% to 50.8%, from 2014 to 2020. Similarly, nonsusceptibility to clindamycin also increased significantly, from 21.0% to 45.8%. In comparison to historical data, the overall rates of GBS infection and antimicrobial resistance have increased and the predominant CPS types have changed. IMPORTANCE This work describes the epidemiology of invasive infections caused by the bacterium group B Streptococcus (GBS) in Alberta, Canada. We show that rates of invasive GBS disease have increased from 2014 to 2020 for both adult disease and late-onset disease in neonates, whereas the rate of early onset disease in neonates has decreased. We also show that the rate of resistance to erythromycin (an antibiotic used to treat GBS) has also increased in this time.

Evaluation of anti-biofouling potential of Viburnum opulus extracts / Evaluación del potencial anti bioincrustante de extractos de Viburnum opulus

In this study the in vitro investigation of the inhibitory effect of ethanol extract of Viburnum opulus L. bark sample on Streptococcus mutans planctonic cells and biofilm has been intended. A Scanning electron microscopy analysis has been performed in order to investigate the inhibitory effect of the extract on Streptococcus mutans biofilms. Furthermore, the Exopolysaccharide and dextran production of this bacteria have been identified in the presence of the extract. It has been found out that the bark extract with the concentration of 2,5 mg/mL is able to inhibit more than 50% of the cells in the different times development phases. According to this, the exopolymeric matrix on the biofilm surface disperses and the Exopolysaccharide and dextran production get lowered in the presence of bark extract compared to the control group. It is considered that this extract can be used as an alternative approach for the new chemotherapeutic strategies against tooth decay.

En este estudio se investigó el efecto inhibitorio in vitro del extracto de etanólico de una muestra de corteza de Viburnum opulus L. en biopelículas de células planctónicas de Streptococcus mutans. Se realizó un análisis de microscopía electrónica de barrido para investigar el efecto inhibitorio del extracto sobre las biopelículas de Streptococcus mutans. Además, se identificó la producción de exopolisacárido y dextrano de esta bacteria en presencia del extracto. Se descubrió que el extracto de corteza con una concentración de 2,5 mg/ml inhibió más del 50% de las células en las diferentes fases de desarrollo. Consecuentemente, la matriz exopolimérica en la superficie de la biopelícula se dispersa y la producción de exopolisacárido y dextrano se reduce en presencia de extracto de corteza en comparación con el grupo de control. Se sugiere que este extracto puede ser usado como un enfoque alternativo para las nuevas estrategias quimioterapéuticas contra la carie dental.

Wine consumers' subjective responses to wine mouthfeel and understanding of wine body.

Wine mouthfeel is considered important for wine quality by experts, while consumers understanding of mouthfeel and the role of wine body in their wine choice is unknown. One experiment determined the influence of intrinsic wine mouthfeel on consumers' wine liking and emotions, and the other, how consumers understand the term wine body. The first experiment used a 2 astringency level×2 body level experimental design. The samples were base wine with; nothing added (control), added xanthan gum (for increased body), added grape seed extract (GSE, for increased astringency), and with both added xanthan gum and GSE. The consumer taste trial (n=112) indicated that wine with increased body did not influence wine liking and emotions; while increased astringency decreased liking and elicited more intense negative emotions. The second experiment examined consumers' knowledge of wine body through an online survey (n=136). Consumers described wine body most frequently using words such as flavour, fullness, and strength. Wine body was therefore understood by consumers predominantly as a holistic multi-sensory perception of flavour. Wine flavour was indicated by consumers to be the most important factor driving purchase decisions followed by balance of flavours and wine body. It is crucial that wine professionals carefully communicate wine characteristics to consumers to prevent possible misunderstandings such as the meaning of wine body and as a result better meet consumer expectations. In future, the term body may benefit from a clearer definition for academic research as well as industry.

Facile green synthesis of baicalein fabricated gold nanoparticles and their antibiofilm activity against Pseudomonas aeruginosa PAO1.

Biofilm formation is one of the major problems associated with chronic diseases and also attributes for the antibiotic resistance in bacteria. In recent times nanoparticles have been utilized to improve the efficacy of the existing antimicrobial and anti-biofilm agents. The ease in functionalization of gold nanoparticles (AuNPs) makes them a potential carrier for antimicrobial agents. However, the use of physical or chemical methods of the production of nanoparticles is expensive, labour intensive and hazardous to ecosystem. On the other hand, the use of plant based compounds serve as an eco-friendly way for the synthesis of nanoparticles with improved biocompatibility and therapeutic applicability. In the present study, phytocompound, baicalein was used as a reducing and capping agent for the synthesis of spherical shape AuNPs. The baicalein decorated gold nanoparticles (BCL-AuNPs) were characterized and evaluated for their anti-biofilm efficacy against Pseudomonas aeruginosa PAO1. The biosynthesized BCL-AuNPs was characterized using UV-Visible spectra, Dynamic Light Scattering (DLS), Fourier transform infrared (FTIR) spectroscopy, Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Diffraction (EDAX), and High Resolution Transmission Electron Microscopy (HR-TEM). The biosynthesized BCL-AuNPs were determined to be spherical in shape with an average size of 26.5 nm. The sub-MIC concentration of BCL-AuNPs exhibited significant anti-biofilm activity against P. aeruginosa PAO1. On treatment with BCL-AuNPs (100 g mL-1), a reduction in biofilm formation by 58.74 ± 5.8% and 76.51 ± 4.27% was observed in microtiter plate assay and tube method, respectively. A significant reduction in exopolysaccharide (EPS) production by 81.29± 2.96% was observed. The swimming and swarming motility were also effectively arrested in presence of BCL-AuNPs. Further, Light microscope and CLSM studies were carried out to examine the effect of BCL-AuNPs on the surface topography and architecture of P. aeruginosa biofilm. Thus, the present study suggests the potential use of BCL-AuNPs in the development of novel therapeutics for the prevention and treatment of biofilm associated chronic infections.

Physicochemical analysis in the evaluation of reconstituted dry emulsion tablets.

The aim of this study was to characterize the formation of emulsions by droplet size analysis and turbidimetry during reconstitution from a solid dosage form, namely from dry emulsion systems, which carry an oil phase for poorly soluble active ingredients. For the dry emulsion systems tablets were prepared either from oil-in-water systems using a freeze-drying process or through direct compression containing the same oil and excipients. The ratios of oil to emulgents and oil to xanthan gum were equal in both methods. In the preparation methods applied, mannitol, erythritol and lactose were used as excipients and mannitol was found to be the most effective excipient based on droplet size reconstitution, turbidimetry and physical properties. Quality control involved testing the physical properties of tablets and characterizing the reconstituted emulsions.

International collaborative study for establishment of the 2nd WHO International Standard for Haemophilus influenzae type b polysaccharide.

In this report we present the results of a collaborative study for the preparation and calibration of a replacement International Standard (IS) for Haemophilus influenzae type b polysaccharide (polyribosyl ribitol phosphate; 5-d-ribitol-(1 â†’ 1)-ß-d-ribose-3-phosphate; PRP). Two candidate preparations were evaluated. Thirteen laboratories from 9 different countries participated in the collaborative study to assess the suitability and determine the PRP content of two candidate standards. On the basis of the results from this study, Candidate 2 (NIBSC code 12/306) has been established as the 2nd WHO IS for PRP by the Expert Committee of Biological Standards of the World Health Organisation with a content of 4.904 ± 0.185mg/ampoule, as determined by the ribose assays carried out by 11 of the participating laboratories.

Calcium binding to S. mutans grown in the presence or absence of sucrose.

Sucrose is the most cariogenic dietary carbohydrate because it is a substrate for insoluble extracellular polysaccharide (IEPS) production in dental biofilms, which can proportionally decrease bacterial density and, consequently, the number of biofilm calcium (Ca) binding sites. Ca bound to bacterial cell walls can be released into the biofilm fluid during a cariogenic challenge, reducing the driving force for mineral dissolution provoked by the pH drop. Thus, we investigated the effect of an IEPS-rich extracellular matrix on bacterial Ca binding after treatment with Ca solutions. Streptococcus mutans Ingbritt 1600 was cultivated in culture broths supplemented with 1.0% sucrose or 0.5% glucose + 0.5% fructose. The IEPS concentration in bacterial pellets was determined after alkaline extraction. Bacterial pellets were treated with 1 mM or 10 mM Ca++ solutions at 37ºC for 10 to 60 min. Ca binding to bacterial pellets, determined after acid extraction using the Arsenazo III reagent, was fast and concentration dependent. Although the IEPS concentration was approximately ten times higher in bacterial pellets cultivated in sucrose as compared to its monossaccharides, bound Ca concentration after Ca treatment was similar in both conditions. These results suggest that IEPS may not influence the amount of Ca bound to reservoirs of dental biofilms.

Calcium binding to S. mutans grown in the presence or absence of sucrose

Sucrose is the most cariogenic dietary carbohydrate because it is a substrate for insoluble extracellular polysaccharide (IEPS) production in dental biofilms, which can proportionally decrease bacterial density and, consequently, the number of biofilm calcium (Ca) binding sites. Ca bound to bacterial cell walls can be released into the biofilm fluid during a cariogenic challenge, reducing the driving force for mineral dissolution provoked by the pH drop. Thus, we investigated the effect of an IEPS-rich extracellular matrix on bacterial Ca binding after treatment with Ca solutions. Streptococcus mutans Ingbritt 1600 was cultivated in culture broths supplemented with 1.0% sucrose or 0.5% glucose + 0.5% fructose. The IEPS concentration in bacterial pellets was determined after alkaline extraction. Bacterial pellets were treated with 1 mM or 10 mM Ca++ solutions at 37ºC for 10 to 60 min. Ca binding to bacterial pellets, determined after acid extraction using the Arsenazo III reagent, was fast and concentration dependent. Although the IEPS concentration was approximately ten times higher in bacterial pellets cultivated in sucrose as compared to its monossaccharides, bound Ca concentration after Ca treatment was similar in both conditions. These results suggest that IEPS may not influence the amount of Ca bound to reservoirs of dental biofilms.

Laboratory assessment of factors affecting soil clogging of soil aquifer treatment systems.

In this study the effect of soil type, level of pre-treatment, ponding depth, temperature and sunlight on clogging of soil aquifer treatment (SAT) systems was evaluated over an eight week duration in constant temperature and glasshouse environments. Of the two soil types tested, the more permeable sand media clogged more than the loam, but still retained an order of magnitude higher absolute permeability. A 6- to 8-fold difference in hydraulic loading rates was observed between the four source water types tested (one potable water and three recycled waters), with improved water quality resulting in significantly higher infiltration. Infiltration rates for ponding depths of 30 cm and 50 cm were higher than 10 cm, although for 50 cm clogging rates were higher due to greater compaction of the clogging layer. Overall, physical clogging was more significant than other forms of clogging. Microbial clogging becomes increasingly important when the particulate concentrations in the source waters are reduced through pre-treatment and for finer textured soils due to the higher specific surface area of the media. Clogging by gas binding took place in the glasshouse but not in the lab, and mechanical clogging associated with particle rearrangement was evident in the sand media but not in the loam. These results offer insight into the soil, water quality and operating conditions needed to achieve viable SAT systems.

Optimization of gluten-free formulations for French-style breads.

The formulation of gluten-free bread, which will be suitable for patients with coeliac disease, was optimized to provide bread similar to French bread. The effects of the presence of hydrocolloids and the substitution of the flour basis by flour or proteins from different sources were studied. The added ingredients were (1) hydrocolloids (carboxymethylcellulose [CMC], guar gum, hydroxypropylmethylcellulose [HPMC], and xanthan gum), and (2) substitutes (buckwheat flour, whole egg powder, and whey proteins). The bread quality parameters measured were specific volume, dry matter of bread, crust color, crumb hardness, and gas cell size distribution. Specific volume was increased by guar gum and HPMC. Breads with guar gum had color characteristics similar to French bread. Hardness decreased with the addition of hydrocolloids, especially HPMC and guar. Breads with guar gum had the most heterogeneous cell size distribution, and guar gum was therefore selected for further formulations. Bread prepared with buckwheat flour had improved quality: an increased specific volume, a softer texture, color characteristics, and gas-cell size distribution similar to French bread. Bread with 1.9% guar gum (w/w, total flour basis) and 5% buckwheat flour (of all flours and substitutes) mimicked French bread quality attributes.

Biosynthesis of emulsan biopolymers from agro-based feedstocks.

AIMS: The need for biocompatible, biodegradable, and versatile biopolymers permeates many fields including environmental and food technology. The goal of the study presented here is to establish the utility of agricultural oils as an inexpensive carbon source to produce materials useful for biomedical materials and offer positive attributes in terms of green chemistry. METHODS AND RESULTS: Structural variants of the complex acylated polysaccharide, emulsan, secreted from Acinetobacter venetianus RAG-1, were biosynthesized in cultures supplemented with agricultural feedstocks to examine the feasibility of conversion of these substrates into value-added biopolymers. Acinetobacter venetianus produced chemically and biologically distinct emulsan variants in culture on soy molasses and tallow oil. These variants possess significant biological function, including macrophage activation and adjuvant activity, in similar range to that observed for the standard emulsan formed on ethanol-fed A. venetianus. CONCLUSIONS: The results indicate that this novel family of biopolymers can be produced in significant quantities from the readily available renewable agricultural feedstocks and the resulting structures and functions can be correlated to the chemistry of these feedstocks. SIGNIFICANCE AND IMPACT OF THE STUDY: The significant quantities of agricultural oils produced annually represent an untapped source for bioconversion to valuable products. The results of this study confirm that the important polymer emulsan can be synthesized from this inexpensive carbon source.

Modification of the Limulus amebocyte lysate assay for the analysis of glucan in indoor environments.

beta-1,3- D-Glucan is a biologically active component mainly from fungi that has been shown in several studies to be related to respiratory health outcomes from damp building exposures. Here, we report the development and application of a method for the analysis of the glucan extracted in 0.5 N NaOH solution making use of an available preparation of Limulus amebocyte lysate (LAL). The method yields reproducible beta-1,3- D-glucan measurements from samples of outdoor air, yeast cells, fungal spore preparations and ragweed pollen, and is more sensitive than competing measurements. The LAL-based measurement compared favourably to that based on size-exclusion chromatography using UV and refractive index detection. Growth conditions of the fungi did not materially change the concentrations of glucan in spores indicating that this is a stable property. Glucan content was proportional to spore surface area; however, some species contain higher relative spore glucan contents.

Application of antibodies for the identification of polysaccharide gum additives in processed foods.

Anti-carbohydrate antibodies with specificities for polysaccharide gums were isolated from the serum of rabbits that were immunized with a solution of the gums and Freund's complete adjuvant. The primary objective was to test an immunological method for the detection of the polysaccharide gums as additives to processed foods. Analysis involved the extraction of food with phosphate buffer and the testing of the extract for a reaction with anti-gum antibodies by the agar diffusion method. Reaction by a specific gum with the homologous antibodies establishes the presence of the gum in the food. The method is a novel application of antibodies. The antibody method is highly specific for a gum and thus possesses advantages over other methods of analysis for polysaccharide gums as additives in processed foods.

Bioengineered emulsans from Acinetobacter calcoaceticusRAG-1 transposon mutants.

Transposon mutants of Acinetobacter calcoaceticus strain RAG-1 were studied in an effort to control fatty acid (FA) substitution patterns of emulsan, a bioemulsifier secreted by the organism. The disrupted genes, involved in the biosynthetic pathways of biotin, histidine, cysteine or purines, influenced the level and types of FAs incorporated into emulsan. The structural variants of emulsan generated by the transposon mutants were characterized for yield, FA content, molecular weight, and emulsification behavior when grown on a series of FAs of different chain lengths from C11 to C18. Yields of emulsan from the transposon mutants were found to be lower than the parent strain and depended on the type of FA used to supplement the growth medium. Mutants 13D (His-) and 52D (Cys-) grown on LB plus C16 or C14, respectively, exhibited enhanced emulsifying activity compared to A. calcoaceticus RAG-1. The presence and composition of long chain FAs on the polysaccharide backbone influenced emulsification behavior: particularly a high mole percentage of C16 (48%) and C18 (42%). The results provide important insight into the bioengineering of bioemulsifier-producing microorganisms and provide a path towards highly tailored novel amphipathic structures to utilize as biodegradable in environmental, biomedical, and personal care applications.

New method for exopolysaccharide determination in culture broth using stirred ultrafiltration cells.

A new method to remove simple carbohydrates from culture broth prior to the quantification of exopolysaccharides (EPS) was developed and validated for the EPS-producing strain, Lactobacillus rhamnosus RW-9595M. This method uses ultrafiltration (UF) in stirred cells followed by polysaccharide detection in the retentate by the phenol-sulfuric acid method. The UF method was compared with a conventional method based on ethanol extraction, dialysis, protein removal by trichloroacetic acid (TCA) and freeze-drying. EPS production during pH-controlled batch fermentations in basal minimum medium, whey permeate (WP). and whey permeate supplemented with yeast extract, minerals and Tween-80 (SWP) was determined by the new UF and conventional methods. EPS recovery by the new method ranged from 83% to 104% for EPS added in the concentration range 40-1,500 mg/l in 0.1 M NaCl solution or culture medium. The UF method was rapid (8 h), accurate and simple, and required only a small sample volume (1-5 ml). A very high maximum EPS production was measured in SWP by both the UF and conventional methods (1,718 and 1,755 mg/l).

[The isolation of the antigenic substances of Mycobacterium tuberculosis and the characteristics of their basic chemical components]. / Vydilennia antyhennykh substantsii mikobakterii tuberkul'ozu ta kharakterystyka ïkh osnovnykh khimichnykh komponentiv.

Different methods of isolation were used when obtaining the antigen determinants of the tuberculosis mycobacteria (tuberculins, polysaccharide, phosphatide). Main chemical components of the antigens were investigated. Most effective and economic methods of obtaining the M. tuberculosis antigens substances were chosen. The using of the obtained antigens in the test systems for diagnostics of tuberculosis discussed.

Characterization of the non-sulphated highly anionic Kurloff cell glycoconjugates by lectin- and immunoblotting: evidence for the presence of alpha (2,8)-linked polysialic acid-bearing molecules.

Urea or guanidine hydrochloride-soluble extracts from highly purified Kurloff cells (KC) radiolabelled in vitro were subjected to DEAE-cellulose chromatography. Among the three anionic peaks obtained, a major and non-sulphated peak (designated as peak IV) strongly affected by glucosamine-labelling and eluted at about 0.3 M NaCl was analyzed. Gel filtration on Sepharose CL4B and 10% SDS-PAGE indicated its heterogeneous size. Peak IV consisted mainly of N-glycans as shown by its susceptibility to tunicamycin. Further insight into its chemical nature was obtained by examining its binding capacity to different lectins and by immunodot analysis. It strongly interacted with concanavalin A (Con A) after dot-blot or Western blotting. A large amount of these glycoproteins is not of the high-mannose type since Galanthus nivalis agglutinin reacted weakly with peak IV. Moreover, bindings to Phaseolus vulgaris and to wheat germ agglutinins suggest the presence of bisecting N-acetylglucosamine residues. Bindings to Sambucus nigra and to Ricinus communis agglutinins, dramatically lessened and increased respectively after desialylation, suggest the presence of Neu5Ac alpha 2,6Gal/GalNAc sequences. The absence of outer sialic acid residues linked alpha 2,3 to galactose was demonstrated following Maackia amurensis agglutinin negativity. The use of poly(alpha 2,8-sialyl) endo-N-acylneuraminidase combined with immunodot procedure with a monoclonal antibody that specifically recognizes alpha 2,8-linked polysialic chains revealed that peak IV contains oligosaccharidic epitopes common to polysialylated neural cell adhesion molecules.

Electrophoretic and chemical characterization of lipopolysaccharides of Vibrio parahaemolyticus.

Lipopolysaccharides (LPSs) isolated from three Kanagawa-positive and three negative strains of Vibrio parahaemolyticus were characterized by using electrophoretic, immunochemical, and chemical methods. The results of this study indicated that the LPSs of all six strains of V. parahaemolyticus examined did not have an O-specific side chain. These V. parahaemolyticus LPSs appeared to have molecular weights similar to that of the rough-type (Ra) LPS of Salmonella typhimurium TV-119 and might just contain lipid A and a core region. However, the microheterogeneity of V. parahaemolyticus LPS observed was greater than that of S. typhimurium LPS. The profile of V. parahaemolyticus LPS consisted of closely spaced triplet or quadruplet bands, but that of S. typhimurium consisted of doublet bands. Slower-moving bands appeared on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels only when large amounts of V. parahaemolyticus LPS were loaded. These bands were proven to be the aggregates of the fastest-moving low-molecular-weight bands by re-electrophoresis. The banding pattern of V. parahaemolyticus LPSs produced on nitrocellulose membranes by immunoblotting indicated that the V. parahaemolyticus LPSs did not have an O-specific side chain. The low ratio of total carbohydrate to lipid A of V. parahaemolyticus LPSs also suggested that they were like rough-type LPS. The mobility and profile of V. parahaemolyticus LPS on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel and its chemical composition were closely related to the serotype of a specific strain but not with the Kanagawa phenomenon.

Observations of the ultrastructure of infected kidney stones.

Struvite stones are formed as the result of urinary tract infection by urease-producing bacteria. Ultrastructural examination of calculi removed from a patient revealed bacteria incorporated throughout the stone matrix. Exopolysaccharide stained by ruthenium red was associated with most of the bacteria, but it represented only a small portion of the organic matrix in the stone. Localised deposits of calcium and phosphorus, components of carbonate-apatite, and magnesium, a struvite component, were detected in close proximity to the cells. Histochemical examinations revealed that several of the gram-negative bacteria within the stone matrix possessed high levels of urease activity. We propose that bacterial slime production, intimately involved in the initiation of stone matrix deposition, is less prominent in mature stones because of the increased incorporation of host-derived mucoproteins and mucopolysaccharides.

[Evaluation of the effectiveness of group-A and -C meningococcal polysaccharide vaccines by molecular parameters]. / Otsenka éffektivnosti meningokokkovykh polisakharidnykh vaktsin grupp A i C po molekuliarnym parametram.

Chromatographic properties of meningococcal polysaccharides and vaccines of serogroups A and C prepared at the G. N. Gabrichevskii Moscow Research Institute of Epidemiology and Microbiology were studied during determination of their molecular parameters by gel filtration on sepharose 4B. It was shown that the character of the polysaccharide elution was not affected by the composition of the eluent and pH changes within the ranges of 6.5-8.5. The ionic strength of the solvent should be at least 0.2 mol/l. Immunochemical methods such as countercurrent and rocket immunoelectrophoresis were standardized and recommended for quantitative evaluation of the content of meningococcal polysaccharides of groups A and C in the eluates. Rocket immunoelectrophoresis was shown to be more accurate and reproducible.