Predictive Evaluation of Microbiological Stability of Soft Drinks with Lycium barbarum L. Stored at Temperature Shifts.

Lycium barbarum L., used in Chinese traditional medicine for centuries, has gained popularity in Europe in the last decade because of its health-promoting properties assigned to phenolic compounds and antioxidant activity. Goji fruits and extracts are often used as ingredients in popular homemade milk cocktails. Within this study, the microbiological stability of the milkshake, with the addition of berries from NingXia Province and their extract, was evaluated using the ComBase® prognostic model. The extraction of dry berries in water at 70 °C for 72 h produced an extract showing radical inhibition of 64.9% and a total phenol content of 63.6 mg g-1. The phenolic compounds with the highest concentrations were in turn: 3-hydroxybenzoic acid, gallic acid, procyanidin B2, and catechin. The milkshake inoculated with the reference B. subtilis was a model for the study of its microbiological stability. Using ComBase®, a microbiological response to the delayed cooling of goji berry extract and the milkshake with the addition of goji berries was predicted and the model's accuracy assessed. The best-performing models were constructed for extract (Bias factor Bf 1.33, Accuracy factor Af 3.43) and milkshake (Bf 1.29, Af 1.65) in a profile simulating delayed refrigeration (22.5 °C-9 °C-23 °C). Despite discrepancies between predicted and observed bacterial growth due to the antimicrobial effect of the derivatives of goji berries, the models were validated as "overpredict", i.e., "fail safe", and may be used to prognose the stability of these products in the given temperature profile.

Fresh and Stored Sugar Beet Roots as a Source of Various Types of Mono- and Oligosaccharides.

Although sugar beets are primarily treated as a source of sucrose, due to their rich chemical composition, they can also be a source of other carbohydrates, e.g., mono- and oligosaccharides. The study focused on both fresh beet roots and those stored in mounds. Our studies have shown that, in addition to sucrose, sugar beet tissue also comprises other carbohydrates: kestose (3.39%) and galactose (0.65%) and, in smaller amounts, glucose, trehalose and raffinose. The acidic hydrolysis of the watery carbohydrates extracts resulted in obtaining significant amounts of glucose (8.37%) and arabinose (3.11%) as well as xylose and galactose and, in smaller amounts, mannose. An HPSEC liquid chromatography study of the molecular mass profile of the carbohydrate compounds present in the beet roots showed alongside the highest percentage (96.53-97.43%) of sucrose (0.34 kDa) the presence of pectin compounds from the araban group and arabinoxylooligosaccharides (5-9 kDa) with a percentage share of 0.61 to 1.87%. On the basis of our research, beet roots can be considered a potential source of carbohydrates, such as kestose, which is classified as fructooligosaccharide (FOS). The results of this study may be helpful in evaluating sugar beets as a direct source of various carbohydrates, or as a raw material for the biosynthesis of fructooligosaccharides (FOS) or galactooligosaccharides (GOS).

Antimicrobial Activities of Plant Extracts against Solanum tuberosum L. Phytopathogens.

The purpose of the study was to select an environmentally friendly plant biopesticide to protect seed potatoes against phytopathogens. The scope included the evaluation of the antimicrobial activities of 22 plant water extracts, 22 water-glycol extracts, and 3 subcritical carbon dioxide extracts using the agar diffusion method against 10 potato phytopathogens. For the most effective extracts, minimal inhibitory concentration (MIC), chemical composition analysis by gas chromatography-mass spectrometry and in situ assays on seed potatoes were performed. Garlic water extract was finally selected as the most effective in phytopathogen growth inhibition, both in vitro and in situ, with MIC values ranging between 6.3-25 mg/mL. 5-Hydroxymethylfurfural was determined to be the main component of this extract (33.24%). Garlic water extract was proposed as a potential biopesticide against potato phytopathogens.

Chemical and Biological Characteristics of Oxytropis pseudoglandulosa Plant of Mongolian Origin.

Oxytropis pseudoglandulosa is used in Mongolian traditional medicine due to its numerous reported health-promoting effects. To date, there are very few scientific reports that describe this species. In this article, its volatile oil composition, lipid extract composition, total phenolic and flavonoid content, antibacterial and allergenic properties are elucidated for the first time. Hexadecanoic acid, fokienol and tricosane were determined as the most notable components of the volatile oil, at 13.13, 11.46 and 5.55%, respectively. Methyl benzoate was shown to be the most abundant component of lipid extract at 40.69, followed by (E)-prop-2-enoic acid, 3-phenyl- and benzenepropanoic acid, at 18.55 and 9.97%. With a TPC of 6.620 mg GAE g-1 and TFC of 10.316 mg QE g-1, the plant extract of O. pseudoglandulosa indicated good antioxidant activity measured by IC50 at 18.761 µg mL-1. Of the 12 tested microorganisms, B. subtilis and S. cerevisiae were the shown to be most susceptible to the plant extract, with MIC at 2.081 and 0.260% (v/v), respectively. Bet v 1-a major birch pollen allergen found in plant-based foods-was determined to be at 192.02 ng g-1 with ELISA. Such a wide spectrum of biological activity indicated by O. pseudoglandulosa lends credence for its application in food industry. Its exerted antioxidant and antimicrobial effects could improve preservation of low-processed food dedicated for consumers afflicted with allergies. Hexadecanoic acid supplemented in foods with dietary plant extracts could add to the potential anti-inflammatory impact. The analysis of lipid makeup suggests O. pseudoglandulosa extract could also be considered as natural pesticide in organic farming.

Chemical and Biological Profile and Allergenicity of Thymus baicalensis Plant of Mongolian Origin.

Thymus baicalensis is a medicinal plant recognized as a traditional Mongolian therapeutic and health-promoting food supplement. The aim of the study was to check the suitability of the tested plant for supporting the treatment of certain diseases. The following study is the first one to showcase the versatile scope of characteristics of T. baicalensis, including its volatile oil composition, polyphenolic composition, lipid composition, phenolic and flavonoid contents, antioxidant activity, antimicrobial properties and ingestive allergenicity. Myrcene, at 26.15%, was shown to be the most abundant component of the volatile oil. Compounds known as inherent components of the Thymus genus: thymol and carvacrol made up only about 0.24% of the extracted oil. As much as 10.11 g kg-1 of polyphenol compounds were identified as derivatives of luteolin-7-O-glucuronide. The lipid extract was found to be rich in palmitic acid (31.05%), while unsaturated fatty acids were not reported. Spectrophotometric determination of the phenols and flavonoids indicated 7.541 mg of gallic acid g-1 and 4.345 mg of quercitin g-1, respectively. The free radical scavenging activity was determined by the 2,2-difenylo-1-pikrylohydrazyl method at IC50 = 206.97 µg mL-1. The extracts also had a strong inhibitory effect on M. flavus and P. fluorescenes bacteria, as well as S. cerevisiae yeasts. The Bet v 1 and profilin allergens in T. baicalensis were reported at 175.17 ng g-1 and 1.66 ng g-1, respectively.

The Impact of Selected Essential Oils Applied to Non-Woven Viscose on Bacteria That Cause Lower Urinary Tract Infections-Preliminary Studies.

Inflammation of the lower urinary tract is a very common problem, which occurs particularly in women. A concept of a biotextronics system for preventive and support treatment of lower urinary tract inflammations was presented. The system includes a non-woven viscose insert for essential oils application. The oils were deposited on the non-woven viscose and incubated in the temperature of 37 °C and served a model for their action in the vapor phase as the element of the biotextronics system. The essential oils used in the research were the following: chamomile (Matricaria chamomilla L.), sage (Salvia officinalis L. and Salvia lavandulaefolia), juniper (Juniperus communis L.), thyme (Thymus vulgaris L.), and mixtures of chamomile oil with oils of each sage species in a 1:1 ratio. The oils were tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus saprophyticus, and Enterococcus faecalis. The best inhibitory effect in vapor phase was noted for chamomile essential oil at the lowest concentration (0.054 µL/cm3). Both mixtures of chamomile and sage acted antagonistically, lowering the antibacterial activity of the individual oils applied solely. Juniper and Salvia officinalis essential oils at the concentrations tested increased the growth of at least one of the bacteria tested. Salvia lavandulaefolia Vahl. essential oil inhibited all bacteria, only at the concentration 0.214 µL/cm3. The thyme oil, at the concentration 0.054 µL/cm3, reduced the growth of all bacterial species tested. Chamomile and thyme essential oils were chosen for further research in the biotextronics pantiliner system.

Clove Oil (Syzygium aromaticum L.) Activity against Alicyclobacillus acidoterrestris Biofilm on Technical Surfaces.

Acidotermophilic bacteria Alicyclobacillus acidoterrestris is one of the main contaminants in the fruit industry forming biofilms which are difficult to remove from the production line by conventional methods. An alternative approach aims for the use of essential oils to prevent Alicyclobacillus biofilm development. The effect of clove essential oil on A. acidoterrestris biofilms on glass and polyvinyl chloride surfaces under static and agitated culture conditions was investigated by atomic force microscopy and the plate count method. The medium-flow and the type of technical surface significantly influenced A. acidoterrestris biofilm. The PVC was colonized in a greater extent comparing to glass. Clove essential oil in 0.05% (v/v) caused 25.1-65.0% reduction of biofilms on the technical surfaces along with substantial changes in their morphology by a decrease in the biofilm: height, surface roughness, and surface area difference. The oil also induced alteration in individual bacterial cells length and visible increase of their roughness. Clove essential oil seems to release EPS from biofilm and thus induce detachment of bacteria from the surface. Due to anti-A. acidoterrestris biofilm activity, the clove oil may be used in the juice industry to hinder a development of A. acidoterrestris biofilms on production surfaces.

Combined Yeast Cultivation and Pectin Hydrolysis as an Effective Method of Producing Prebiotic Animal Feed from Sugar Beet Pulp.

An effective and ecological method for liberation of pectin-derived oligosaccharides (POS) from sugar beet pulp (SBP) was developed using enzymatic and microorganism-mediated biomass conversion. The POS may be applied in the production of prebiotic feed additives. Various yeast strains were screened for their capacity for protein synthesis and monosaccharide assimilation. Combined yeast cultivation and pectin hydrolysis were found to be an effective method of producing prebiotics. Separate enzymatic hydrolysis and fermentation of SBP resulted in the release of 3.6 g of POS per 100 g d.w., whereas the yield of POS acquired after the combined process was 17.9% higher, giving 4.2 g of POS per 100 g d.w. Introducing the yeast into the process improved hydrolysis performance due to lower enzyme inhibition by mono- and disaccharides. The prebiotic effect of the POS was assessed by in vitro fermentation using individual cultures of gastrointestinal bacteria. The POS in the SBP hydrolysate effectively promoted the growth of lactobacilli and bifidobacteria. A large increase in adherence to Caco-2 cells in the presence of POS was noted for beneficial Lactobacillus brevis strains, whereas pathogenic bacteria and yeast (C. albicans, C. lusitanie, C. pelliculosa), responsible for infections in breeding animals, showed much weaker adhesion.

Effect of Clove and Thyme Essential Oils on Candida Biofilm Formation and the Oil Distribution in Yeast Cells.

Candida biofilm structure is particularly difficult to eradicate, since biofilm is much more resistant to antifungal agents than planktonic cells. In this context, a more effective strategy seems to be the prevention of biofilm formation than its eradication. The aim of the study was to examine whether the process of initial colonization of materials (glass, polyethylene terephthalate, polypropylene) by food-borne Candida sp. can be impeded by clove and thyme essential oils, used at their minimal inhibitory concentrations. In the presence of clove oil, 68.4-84.2% of the yeast tested showed a statistically significant reduction in biofilm formation, depending on the material. After treatment with thyme oil, statistically significant decrease in biofilm cell numbers was observed for 63.2-73.7% of yeasts. Confocal laser scanning microscopy showed diverse compounds of clove and thyme oils that were disparately located in C. albicans cell, on a cell wall and a cell membrane, in cytoplasm, and in vacuoles, depicting the multidirectional action of essential oils. However, essential oils that were used in sub-inhibitory concentration were sequestrated in the yeast vacuoles, which indicate the activation of Candida defense mechanisms by cell detoxification. Clove and thyme essential oils due to their anti-biofilm activity can be efficiently used in the prevention of the tested abiotic surfaces colonization by Candida sp.

Candida albicans Impairments Induced by Peppermint and Clove Oils at Sub-Inhibitory Concentrations.

Members of Candida species cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. In order to prevent from Candida sp. development, essential oils are more and more frequently applied, due to their antifungal activity, low toxicity if used appropriately, and biodegrability. The aim of the study was to characterize the early alterations in Candida albicans metabolic properties in relation to proteins and chromosomal DNA profiles, after treatment with peppermint and clove oils at sub-inhibitory concentrations. The yeasts were affected by the oils even at a concentration of 0.0075% v/v, which resulted in changes in colony morphotypes and metabolic activities. Peppermint and clove oils at concentrations ranging from 0.015× MIC (minimal inhibitory concentration) to 0.5× MIC values substantially affected the enzymatic abilities of C. albicans, and these changes were primarily associated with the loss or decrease of activity of all 9 enzymes detected in the untreated yeast. Moreover, 29% isolates showed additional activity of N-acetyl-ß-glucosaminidase and 14% isolates-α-fucosidase in comparison to the yeast grown without essential oils addition. In response to essential oils at 0.25-0.5× MIC, extensive changes in C. albicans whole-cell protein profiles were noted. However, the yeast biochemical profiles were intact with the sole exception of the isolate treated with clove oil at 0.5× MIC. The alterations were not attributed to gross chromosomal rearrangements in C. albicans karyotype. The predominantly observed decrease in protein fractions and the yeast enzymatic activity after treatment with the oils should be considered as a phenotypic response of C. albicans to the essential oils at their sub-inhibitory concentrations and may lead to the reduction of this yeast pathogenicity.

Selected Essential Oils as Antifungal Agents Against Antibiotic-Resistant Candida spp.: In Vitro Study on Clinical and Food-Borne Isolates.

Candida spp. cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. As a result of the increasing antibiotic resistance among pathogenic yeasts, the interest in alternative agents of antifungal activity is growing. This study evaluated the antimicrobial activity of selected essential oils (EOs) against Candida clinical and food-borne strains, including antibiotic-resistant isolates, in relation to yeast cell surface hydrophobicity (CSH). Candida strains showed different range of susceptibility to tea tree, thyme, peppermint, and clove oils, and peppermint oil demonstrated the lowest anticandidal activity with minimal inhibitory concentrations (MICs) of 0.03-8.0% v/v. MIC values for thyme and clove oils ranged from 0.03% to 0.25% v/v, and for tea tree oil-from 0.12% to 2.0% v/v. The exception was Candida tropicalis food-borne strain, the growth of which was inhibited after application of EOs at concentration of 8% v/v. Due to diverse yeast susceptibility to EOs, isolates were divided into five clusters in a principal component analysis model, each containing both clinical and food-borne strains. Hydrophobic properties of yeast were also diversified, and 37% of clinical and 50% of food-borne strains exhibited high hydrophobicity. The study indicates high homology of clinical and food-borne Candida isolates in relation to their susceptibility to anticandidal agents and hydrophobic properties. The susceptibility of yeasts to EOs could be partially related to their CSH. High antifungal activity of examined EOs, also against antibiotic-resistant isolates, indicates their usefulness as agents preventing the development of Candida strains of different origin.

The effect of thyme and tea tree oils on morphology and metabolism of Candida albicans.

Members of Candida species cause significant problems in medicine and in many industrial branches also. In order to prevent from Candida sp. development, essential oils are more and more frequently applied as natural, non-toxic, non-pollutive and biodegradable agents with a broad spectrum of antimicrobial activity. The aim of the research was to determine changes in morphology and metabolic properties of Candida albicans in the presence of thyme and tea tree oils. Changes of enzymatic activity of isolates were observed in the presence of both tested essential oils, and they were primarily associated with loss or decrease of activity of all enzymes detected for control. Furthermore, only for 3 out of 11 isolates additional activity of N-acetyl-ß-glucosaminidase, α-mannosidase, α-fucosidase and trypsin was detected. Vivid changes in biochemical profiles were found after treatment with tea tree oil and they were related to loss of ability to assimilate D-xylose, D-sorbitol and D-trehalose. The main differences in morphology of isolates compared to the control strain concerned formation of pseudohyphae structures. Both examined essential oils caused changes in cell and colony morphology, as well as in the metabolism of Candida albicans. However, the extent of differences depends on the type and concentration of an essential oil. The most important finding is the broad spectrum of changes in yeast enzymatic profiles induced by thyme and tea tree oils. It can be supposed that these changes, together with loss of ability to assimilate saccharides could significantly impact Candida albicans pathogenicity.