A fibre and phenolic-rich flour from Isabel grape by-products with stimulatory effects on distinct probiotics and beneficial impacts on human colonic microbiota in vitro.

This study evaluated the effects of a fibre and phenolic-rich flour (IGF) prepared from Isabel grape by-products on the growth and metabolism of different probiotics and distinct bacterial populations part of the human intestinal microbiota during an in vitro colonic fermentation. IGF was submitted to simulated gastrointestinal digestion before use in the experiments. IGF favoured the growth of the probiotics Lactobacillus acidophilus La-05, L. casei L-26 and Bifidobacterium lactis Bb-12, with viable counts of >7 log CFU per ml, as well as caused decreases in pH values and increases in organic acid production in the growth medium during 48 h of cultivation. IGF increased the population of beneficial micro-organisms forming the human intestinal microbiota, particularly Lactobacillus spp., decreased the pH values, and increased the lactic acid and short-chain fatty acid (acetic, butyric and propionic acids) production during 24 h of in vitro colonic fermentation. These results indicate the potential prebiotic effects of IGF, which should represent a novel sustainable added-value ingredient with functional properties and gut-health benefits.

Study of the proximate and mineral composition of different Nigerian yam chips, flakes and flours.

Yam (Dioscorea spp) is an essential tuber crop for hundreds of millions of people in many African, Asian and South American countries. Considering in particular Southwest Nigeria, chips, flakes and flours are amongst the most common shelf-stable traditionally-processed yam products. This paper reports a systematic study on the proximate (moisture, protein, carbohydrate, fibre, fat, ash and gross energy) and mineral composition of these three food commodities sold in Nigerian markets. Results showed no significant differences in the moisture, crude protein and fibre content of all samples (10.0-12.3, 2.7-4.3 and 1.3-2.0 wt%, respectively). Gross energy was also comparable for all yam derived food items (between 3300 and 3507 kcal/kg), contradicting the common belief that yam flakes have lower nutritional value than chips and flours. Considering the mineral composition, Ca, Mg, P and K were the predominant macronutrients. Micronutrients such as Zn, Co, Mn and Cu were also detected. Significant differences existed between products, and their various sources (markets). Principal component analysis showed a direct correlation between ash content of the samples and the assessed macronutrients, irrespective of the market, or the seller of the commodities. This study confirmed that yam derived food stuffs have an adequate nutritional composition, irrespective of their form and/or origin.

Chitosan as an effective inhibitor of multidrug resistant Acinetobacter baumannii.

Over the last two decades worldwide levels of antibiotic resistance have risen leading to the appearance of multidrug resistant microorganisms. Acinetobacter baumannii is a known skin pathogen which has emerged as a major cause of nosocomial outbreaks due to its capacity to colonize indwelling medical devices and natural antibiotic resistance. With chitosan being an effective antimicrobial agent against antibiotic resistant microorganisms, the aim of this work was to access its potential as an alternative to traditional antimicrobials in the management of A. baumannii growth. What the results showed was that both chitosan MW's tested were active upon A. baumannii's planktonic and sessile growth. For planktonic growth MICs and MBCs were obtained at relatively low concentrations (0.5-2mg/mL) while for sessile growth chitosan proved to be an effective inhibitor of A. baumannii's adhesion and biofilm formation. Considering these results chitosan shows a high potential for control of A. baumannii infections.

Chitosan nanoparticles as alternative anti-staphylococci agents: Bactericidal, antibiofilm and antiadhesive effects.

Chitosan is a biocompatible, bioactive, non-toxic polymer that due to these characteristics has been widely used as a carrier for targeted delivery of bioactive molecules. In recent years, and considering that chitosan has a strong antimicrobial potential, the scientific community's focus has shifted onto the possible antimicrobial activity of chitosan nanoparticles. With this in mind, the aim of this work was to produce low molecular weight chitosan nanoparticles, through the ionic gelation method and characterize their potential biological activity against three staphylococci (MSSA, MRSA and MRSE) in planktonic and sessile environments. The chitosan nanoparticles produced had an average size of 244±12nm, an average charge of 17.3±1.4mV and had a MIC of 1.25mg/mL for all tested microorganisms. Bactericidal activity was only registered for MSSA and MRSA with the time-inhibition curves showing bactericidal activity within 1h. Assays regarding chitosan nanoparticles' impact upon sessile populations showed that they were effective in preventing MRSE adhesion and highly effective in reducing MRSA and MSSA biofilm formation.

Insights into chitosan antibiofilm activity against methicillin-resistant Staphylococcus aureus.

AIMS: Chitosan is a natural compound that has been validated as a viable antimicrobial agent against Staphylococcus aureus. With this work we sought to evaluate the planktonic and sessile sensitivity of methicillin-resistant S. aureus to chitosan's activity and evaluate if methicillin-resistant S. aureus (MRSA) would be more or less sensitive to chitosan's activity than methicillin-sensitive S. aureus (MSSA). METHODS AND RESULTS: A group comprised of reference strains and clinical multiresistant isolates of MSSA and MRSA were used. Methicilin resistance effect upon chitosan activity was assessed in planktonic setting and in different phases of sessile colonization, namely adhesion, biofilm formation and mature biofilm through biomass and metabolism inhibition. The results obtained showed that S. aureus methicillin resistance mechanism did not impair chitosan's activity as the highest bacterial susceptibility was registered for MRSA. Chitosan was highly effective in inhibiting MSSA and MRSA strains in both planktonic and sessile settings with biofilm inhibition percentages reaching as high as 90% for MRSA. CONCLUSIONS: Staphylococcus aureus methicillin resistance did not impair chitosan's antimicrobial and antibiofilm activities and MRSA and MSSA were inhibited both in planktonic and sessile settings at low concentrations with great efficacy. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering the obtained results chitosan shows potential as an alternative for the control of biofilm-related recalcitrant MRSA infections.

Effect of preparation and processing conditions on UV absorbing properties of hydroxyapatite-Fe2O3 sunscreen.

The development of innovative, safe and non-photocatalytic sunscreens is urgently needed, as it is essential to have sunscreen filters offering appropriate UV protection without damaging the environment and/or generating free radicals when in contact with the skin. Hydroxyapatite (Ca10(PO4)6(OH)2, HAp) when substituted with iron has UV protection properties and is not photocatalytic; HAp was used to make a sunscreen filter by treating cod fish bones in an iron-containing solution, and then calcining them at 700°C. Here we present a systematic and advanced study on this material, to obtain a sunscreen with improved UV absorbing properties. Bones were treated with three different iron salts - Fe(II) chloride, Fe(II) lactate and Fe(III) nitrate - under various pH conditions. Results showed that Fe(II) chloride in basic pH led to the most effective iron inclusion. High energy ball milling or ultrasound were investigated to increase surface area and corresponding UV absorption; high energy ball milling treatment led to the best optical properties. The optimum powders were used to formulate UV protection creams, which showed Sun Protection Factor (SPF) values significantly superior to the control cream (up to 4.1). Moreover the critical wavelength (λcrit) was >370nm (388-389nm) and UVA/UVB ratios were very close to 1. With these properties these sunscreens can be classified as broad UV protectors. Results also showed that combining these powders with other sunscreens (i.e. titanium dioxide), a synergic effect between the different components was also observed. This investigation showed that HAp-based sunscreens of marine origin are a valid alternative to commercial products, safe for the health of the customers and, being non-photocatalytic, do not pose a threat to the environment.

Antimicrobial, antiadhesive and antibiofilm activity of an ethanolic, anthocyanin-rich blueberry extract purified by solid phase extraction.

AIMS: The present work aimed to characterize the impact of an anthocyanin-rich blueberry extract upon the growth, adhesion and biofilm formation of several pathogens including some multiresistant bacteria. METHODS AND RESULTS: A group comprised of reference strains and clinical multiresistant isolates of Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Acinetobacter baumannii and Staphylococcus aureus, were used to screen for antimicrobial activity. Microbial growth was determined through the measurement of the optical density while adhesion and biofilm formation was determined using the standard crystal violet staining procedure. The results showed that, while blueberry extract was only effective in hindering the growth of Staph. aureus and E. coli, it was capable of significantly inhibiting biofilm formation and bacterial adhesion for all micro-organisms tested. CONCLUSIONS: The extract demonstrated a considerable potential as a natural, alternative antimicrobial capable of either interfering with microbial growth or hamper the adhesion to surfaces, with Staph. aureus proving to be the most susceptible micro-organism. SIGNIFICANCE AND IMPACT OF THE STUDY: The overall study demonstrates the potential of anthocyanin extracts as natural effective alternative antimicrobial agents. Additionally, the extract's capacity to reduce adhesion without reducing bacterial growth reduces the likeliness of resistance development while reducing the probability of infection.

Hydroxyapatite-based materials of marine origin: a bioactivity and sintering study.

Single phase hydroxyapatite (HAp) and biphasic material hydroxyapatite/ß-tricalcium phosphate (HAp/ß-TCP) were obtained from a marine source (Atlantic cod fish bones). Here we report a study on the biological properties of these materials, including cytotoxicity, bioactivity and haemocompatibility. Results showed that the materials are not cytotoxic, neither in their powder nor in pellet form; indeed growth of Saos-2 cells was comparable to that of commercial. The haemolysis rate was lower than 2%; hence the materials can be classified as non-haemolytic. Moreover, when immersed in Simulated Body Fluid (SBF), crystal formation was observed on the surface of both materials. The sintering behaviour of the samples was also studied; both powders showed very high sinterability (density higher than 95% of the theoretical value). Overall, these results confirm the suitability of these materials for biomedical applications.

Chitosan mouthwash: toxicity and in vivo validation.

A previous study showed that a chitosan mouthwash would be a valid alternative to current mouthwashes as it demonstrated, in vitro, significantly higher antibiofilm activity than two commercial mouthwashes. As such, the aim of this work was to verify the safety of the developed product and to validate, in vivo, the biological activity ascertained in vitro. Chitosan mouthwash safety was evaluated through Ames, MTT and V79 chromosomal aberration assay while antimicrobial activity was evaluated through in vivo assays. The results showed that the chitosan mouthwash was safe, presenting lower cytotoxicity than a commercial mouthwash, and that it effectively reduced viable counts of Streptococcus spp. and Enterococcus spp. by ca. 5.5 log of CFU. Furthermore, in direct comparison with a commercial mouthwash the chitosan mouthwash possessed significantly higher antimicrobial activity. The conjunction of these results proves that the chitosan mouthwash is a safe, effective, natural alternative to the existent chemical mouthwashes.

A hydroxyapatite-Fe2O3 based material of natural origin as an active sunscreen filter.

The use of sunscreens as protective barriers against skin damage and cancer, by absorbing harmful UVA and UVB rays, is becoming an increasingly important issue. Such products are usually based on TiO2 or ZnO, although both Fe2O3 and hydroxyapatite (Ca10(PO4)6(OH)2, HAp) doped with metal ions have been reported as being ultraviolet (UV) absorbing materials. HAp is the main component of bone; it is, therefore, highly biocompatible. In the present work, an iron-doped HAp-based material, containing both Fe ions substituted into the HAp structure and iron oxide in hematite (α-Fe2O3) form, was successfully developed from waste cod fish bones. This was achieved through a simple process of treating the bones in a Fe(ii) containing solution, followed by heating at 700 °C. The material showed good absorption in the whole UV range and did not form radicals when irradiated. The sunscreen cream formulated with this material could be used as a broad sunscreen protector (λcrit > 370 nm), showing high absorption both in the UVA and UVB ranges. Because of its absorption properties it would be classified as 5 star protection according to the Boots UVA star rating system. The cream is also photostable, and does not cause irritation or erythema formation when in contact with the human skin. These results show that a food by-product such as fish bones could be converted into a valuable product, with potential applications in health care and cosmetics. This is the first time a HAp-based sunscreen cream has been developed and validated as a proof of concept.

A comprehensive study into the impact of a chitosan mouthwash upon oral microorganism's biofilm formation in vitro.

Modern dentistry emphasizes the importance of dental plaque control to improve oral health. To that end the development of oral care formulations has been geared toward the incorporation of antiplaque agents that may play a crucial role in oral health maintenance. In later years the research into antiplaque agents has led to the discovery of compounds with significant capability to affect biofilm formation. Among these compounds was chitosan, a polysaccharide which showed great ability to interfere with Streptococcus mutans biofilm formation. As such the aim of this work was to incorporate chitosan into a mouthwash matrix and assess its effect upon biofilm formation of oral microorganisms. This assessment was performed via study of the impact the mouthwash upon microbial adherence, biofilm formation and mature biofilms. Additionally, the action of the chitosan mouthwash was compared with two commercially available mouthwashes. The results here obtained show that only the chitosan containing mouthwash was capable of interfering with all microorganisms' adherence, biofilm formation and mature biofilms while at the same time showing vastly superior activity than both commercial mouthwashes assayed. As such a chitosan mouthwash shows great potential as a natural and efficient alternative to traditional mouthwashes.

A novel direct contact method for the assessment of the antimicrobial activity of dental cements.

Dental cements are a crucial part of the odontological treatment, however, due to the hazardous nature and reduced biological efficiency of some of the used materials, newer and safer alternatives are needed, particularly so those possessing higher antimicrobial activity than their traditional counterparts. The evaluation of the antimicrobial properties of solid and semi-solid antimicrobials, such as dental cements and gels, is challenging, particularly due to the low sensitivity of the current methods. Thus, the main aim of this study was the evaluation of the antimicrobial activity of a novel chitosan containing dental cement while simultaneous assessing/validating a new, more efficient, method for the evaluation of the antimicrobial activity of solid and gel like materials. The results obtained showed that the proposed method exhibited a higher sensitivity than the standard 96 well microtiter assay and allowed the determination of bactericidal activity. Additionally, it is interesting to note that the chitosan containing cement, which presented higher antimicrobial activity than the traditional zinc oxide/eugenol mix, was capable of inducing a viable count reduction above 5 log of CFU for all of the studied microorganisms.

Study of the effects of chitosan upon Streptococcus mutans adherence and biofilm formation.

The main aim of this work was to access the potential use of high and low molecular weight chitosans as potential oral antimicrobials, particularly as antibiofilm agents. Chitosan's interference with Streptococcus mutans capability to adhere and form biofilms was assessed. Additionally the effect upon mature and polymicrobial biofilms was also evaluated. The results obtained showed that chitosan was capable of interfering with S. mutans adhesion and primary biofilm formation. This action was observed up to a week with little to none decrease in efficiency. In addition chitosan was capable of inhibiting biofilms formed by two microorganisms and was capable of acting upon mature biofilms leading to significant reductions (94%) in biofilm survival. However clear statistical differences (p < 0.05) were registered in all assays with, in most assays, HMw chitosan presenting higher efficiency than LMw chitosan. Considering this results chitosan's potential as a valid alternative to traditional antimicrobials in oral health it's evident.

Extraction and characterisation of apatite- and tricalcium phosphate-based materials from cod fish bones.

Apatite- and tricalcium phosphate-based materials were produced from codfish bones, thus converting a waste by-product from the food industry into high added-valued compounds. The bones were annealed at temperatures between 900 and 1200 °C, giving a biphasic material of hydroxyapatite and tricalcium phosphate (Ca10(PO4)6(OH)2 and ß-Ca(PO4)3) with a molar proportion of 75:25, a material widely used in biomedical implants. The treatment of the bones in solution prior to their annealing changed the composition of the material. Single phase hydroxyapatite, chlorapatite (Ca10(PO4)6Cl2) and fluorapatite (Ca10(PO4)6F2) were obtained using CaCl2 and NaF solutions, respectively. The samples were analysed by several techniques (X-ray diffraction, infrared spectroscopy, scanning electron microscopy and differential thermal/thermogravimetric analysis) and by elemental analyses, to have a more complete understanding of the conversion process. Such compositional modifications have never been performed before for these materials of natural origin to tailor the relative concentrations of elements. This paper shows the great potential for the conversion of this by-product into highly valuable compounds for biomedical applications, using a simple and effective valorisation process.

Evaluation and insights into chitosan antimicrobial activity against anaerobic oral pathogens.

The objective of this study was to assess the antimicrobial capability of non-chemically altered chitosan as an alternative to traditional antimicrobials used in the treatment of oral infections. The action mechanism of chitosan was also ascertained. High and low molecular weight chitosan showed antimicrobial activity at low concentrations for all tested bacteria with the MICs varying between 1 and 7 mg/ml with a drop of efficacy relatively to the action of LMW chitosan. In addition chitosan showed also to be an effective bactericidal presenting bactericidal effect within 8 h at the latest. Additionally the evaluation of chitosan's action mechanism showed that both MWs acted upon the bacterial cell wall and were not capable of interacting with the intracellular substances, as showed by the inefficacy obtained in the flocculation assay.