Investigation on Fermented Milk Quality after the Addition of Flaxseed Mucilage and the Use of Lactobacillus delbrueckii subsp. bulgaricus and Lactiplantibacillus plantarum AG9.

BACKGROUND: Flaxseed mucilage (FSM) is one of the healthy components of flaxseed. FSM is an example of a material that can be used in the food, cosmetic, and pharmaceutical industries due to its rheological properties. FSM consists mainly of two polysaccharides, arabinoxylan, and rhamnogalacturonan I, and it also contains protein components and minerals. The prospect of using FSM in food is due to its gelling, water binding, emulsifying, and foaming properties. In addition, valuable natural sources of phenolic compounds such as lignans, phenolic acids, flavonoids, phenylpropanoids, and tannins are partially extracted from flaxseed in FSM. These antioxidant components have pharmacological properties, including anti-diabetic, anti-hypertensive, immunomodulatory, anti-inflammatory and neuroprotective properties. A combination of FSM and lactobacilli in dairy foods can improve their functional properties. This study aimed to develop dairy products by adding of FSM and using two lactic acid bacteria (LAB). FSM (0.2%) was used as an ingredient to improve both the texture and antioxidant properties of the product. METHODS: Skim milk was fermented with 0.2% flaxseed mucilage using Lactobacillus delbrueckii subs. bulgaricus and the probiotic Lactiplantibacillus plantarum AG9. The finished fermented milk products were stored at 4 °C for 14 days. Quantitative chemical, textural, and antioxidant analyses were carried out. RESULTS: Adding 0.2% FSM to the dairy product stimulated the synthesis of lactic acid. FSM increased the viscosity and water-holding capacity of L. bulgaricus or L. bulgaricus/L. plantarum AG9 fermented milk products. Combining these starter strains with FSM promoted the formation of a hard, elastic, resilient casein matrix in the product. When only L. plantarum AG9 was used for the fermentation, the dairy product had a high syneresis and a low viscosity and firmness; such a product is inferior in textural characteristics to the variant with commercial L. bulgaricus. The addition of FSM improved the textural properties of this variant. The use of L. plantarum AG9 and FSM makes it possible to obtain a fermented milk product with the highest content of polyphenolic compounds, which have the highest antioxidant properties and stimulate lipase and α-glucosidase inhibitor synthesis. Combining of L. bulgaricus and L. plantarum AG9 in the starter (20% of the total mass of the starter) and adding of 0.2% FSM is the optimal combination for obtaining a dairy product with high textural and antioxidant properties. CONCLUSIONS: The physicochemical properties (viscosity, syneresis, water holding capacity, texture) and antioxidant properties of fermented milk were improved. In the future, as part of the work to investigate the functional properties of dairy products with FSM, studies will be conducted using in in vivo models.

Potential risk assessment and occurrence characteristic of heavy metals based on artificial neural network model along the Yangtze River Estuary, China.

Pollution from heavy metals in estuaries poses potential risks to the aquatic environment and public health. The complexity of the estuarine water environment limits the accurate understanding of its pollution prediction. Field observations were conducted at seven sampling sites along the Yangtze River Estuary (YRE) during summer, autumn, and winter 2021 to analyze the concentrations of seven heavy metals (As, Cd, Cr, Pb, Cu, Ni, Zn) in water and surface sediments. The order of heavy metal concentrations in water samples from highest to lowest was Zn > As > Cu > Ni > Cr > Pb > Cd, while that in surface sediments samples was Zn > Cr > As > Ni > Pb > Cu > Cd. Human health risk assessment of the heavy metals in water samples indicated a chronic and carcinogenic risk associated with As. The risks of heavy metals in surface sediments were evaluated using the geo-accumulation index (Igeo) and potential ecological risk index (RI). Among the seven heavy metals, As and Cd were highly polluted, with Cd being the main contributor to potential ecological risks. Principal component analysis (PCA) was employed to identify the sources of the different heavy metals, revealing that As originated primarily from anthropogenic emissions, while Cd was primarily from atmospheric deposition. To further analyze the influence of water quality indicators on heavy metal pollution, an artificial neural network (ANN) model was utilized. A modified model was proposed, incorporating biochemical parameters to predict the level of heavy metal pollution, achieving an accuracy of 95.1%. This accuracy was 22.5% higher than that of the traditional model and particularly effective in predicting the maximum 20% of values. Results in this paper highlight the pollution of As and Cd along the YRE, and the proposed model provides valuable information for estimating heavy metal pollution in estuarine water environments, facilitating pollution prevention efforts.

Typical antibiotic resistance genes and their association with driving factors in the coastal areas of Yangtze River Estuary.

The massive use of antibiotics has led to the escalation of microbial resistance in aquatic environment, resulting in an increasing concern regarding antibiotic resistance genes (ARGs), posing a serious threat to ecological safety and human health. In this study, surface water samples were collected at eight sampling sites along the Yangtze River Estuary. The seasonal and spatial distribution patterns of 10 antibiotics and target genes in two major classes (sulfonamides and tetracyclines) were analyzed. The findings indicated a high prevalence of sulfonamide and tetracycline resistance genes along the Yangtze River Estuary. Kruskal-Wallis analysis revealed significant seasonal variations in the abundance of all target genes. The accumulation of antibiotic resistance genes in the coastal area of the Yangtze River Estuary can be attributed to the influence of urban instream runoff and the discharge of effluents from wastewater treatment plants. ANISOM analysis indicated significant seasonal differences in the microbial community structure. VPA showed that environmental factors contribute the most to ARG variation. PLS-PM demonstrate that environmental factors and microbial communities pose direct effect to ARG variation. Analysis of driving factors influencing ARGs in this study may shed new insights into the mechanism of the maintenance and propagation of ARGs.

Predictions of heavy metal concentrations by physiochemical water quality parameters in coastal areas of Yangtze river estuary.

Due to the degradation-resistant and strong toxicity, heavy metals pose a serious threat to the safety of water environment and aquatic ecology. Rapid acquisition and prediction of heavy metal concentrations are of paramount importance for water resource management and environmental preservation. In this study, heavy metal concentrations (Cr, Ni, Cu, Pb, Zn, Cd) and physicochemical parameters of water quality including Temperature (Temp), pH, Oxygen redox potential (ORP), Dissolved oxygen (DO), Electrical conductivity (EC), Electrical resistivity (RES), Total dissolved solids (TDS), Salinity (SAL), Cyanobacteria (BGA-PE), and turbidity (NTU) were measured at seven stations in the Yangtze river estuary. Principal Component Analysis (PCA) and Spearman correlation analysis were employed to analyze the main factors and sources of heavy metals. Results of PCA revealed that the main sources of Cr, Ni, Zn, and Cd were steel industry wastewater, domestic and industrial sewage, whereas shipping and vessel emissions were typically considered sources of Pb and Cu. Spearman correlation analysis identified Temp, pH, ORP, EC, RES, TDS, and SAL as the key physicochemical parameters of water quality, exhibiting the strongest correlation with heavy metal concentrations in sediment and water samples. Based on these results, multiple linear regression as well as non-linear models (SVM and RF) were constructed for predicting heavy metal concentrations. The results showed that the results of the nonlinear model were more suitable for predicting the concentrations of most heavy metals than the linear model, with average R values of the SVM test set and RF test set being 0.83 and 0.90. The RF model showed better applicability for simulating the concentration of heavy metals along the Yangtze river estuary. It was demonstrated that non-linear research methods provided efficient and accurate predictions of heavy metal concentrations in a simple and rapid manner, thereby offering decision-making support for watershed managers.

Repression of Staphylococcus aureus and Escherichia coli by Lactiplantibacillus plantarum Strain AG10 in Drosophila melanogaster In Vivo Model.

Probiotic bacteria exhibiting antagonistic activities against pathogenic bacteria are widely considered as potential options for the prevention and treatment of various infectious diseases and represent potential substitutes of antibiotics. Here we show that the L. plantarum AG10 strain represses the growth of Staphylococcus aureus and Escherichia coli in vitro and diminishes their negative effects in vivo in a Drosophila melanogaster model of survival on embryonic (larvae) and pupa stages. In an agar drop diffusion test, L. plantarum AG10 exhibited antagonistic properties against Escherichia coli, Staphylococcus aureus, Serratia marcescens and Pseudomonas aeruginosa, and repressed the growth of E. coli and S. aureus during milk fermentation. In a Drosophila melanogaster model, L. plantarum AG10 alone did not provide any significant effect, either during the embryonic stage or during further development of the flies. Despite this, it was able to restore the viability of groups infected with either E. coli and S. aureus, almost to the level of non-treated control at all stages of development (larvae, pupa and adult). Moreover, in the presence of L. plantarum AG10, pathogens-induced mutation rates and recombination events reduced 1.5-2-fold. The genome of L. plantarum AG10 was sequenced and deposited at NCBI under the accession number PRJNA953814 and consists of annotated genome and raw sequence data. It consists of 109 contigs and is 3,479,919 bp in length with a GC content of 44.5%. The analysis of the genome has revealed considerably few putative virulence factors and three genes responsible for the biosynthesis of putative antimicrobial peptides, with one of them exhibiting a high probability of antimicrobial properties. Taken together, these data allow the suggestion that the L. plantarum AG10 strain is promising for use in both dairy production and probiotics as a preservative from foodborne infections.

Spatiotemporal distributions of sulfonamide and tetracycline resistance genes and microbial communities in the coastal areas of the Yangtze River Estuary.

In this paper, water and sediments were sampled at eight monitoring stations in the coastal areas of the Yangtze River Estuary in summer and autumn 2021. Two sulfonamide resistance genes (sul1 and sul2), six tetracycline resistance genes (tetM, tetC, tetX, tetA, tetO, and tetQ), one integrase gene (intI1), 16 S rRNA genes, and microbial communities were examined and analyzed. Most resistance genes showed relatively higher abundance in summer and lower abundance in autumn. One-way analysis of variance (ANOVA) showed significant seasonal variation of some ARGs (7 ARGs in water and 6 ARGs in sediment). River runoff and WWTPs are proven to be the major sources of resistance genes along the Yangtze River Estuary. Significant and positive correlations between intI1 and other ARGs were found in water samples (P < 0.05), implying that intI1 may influence the spread and propagation of resistance genes in aquatic environments. Proteobacteria was the dominant phylum along the Yangtze River Estuary, with an average proportion of 41.7%. Redundancy analysis indicated that the ARGs were greatly affected by temperature, dissolved oxygen, and pH in estuarine environments. Network analysis showed that Proteobacteria and Cyanobacteria were the potential host phyla for ARGs in the coastal areas of the Yangtze River Estuary.

Seasonal changes in blood rheology in little ground squirrels.

The annual cycle of small ground squirrels (Spermophilus pygmaeus) consists of 2 periods: wakefulness (spring-autumn) and hibernation (winter). During the active period, ground squirrels breed (spring), accumulate fat reserves (summer) and prepare for hibernation (autumn). We suppose that the rheological properties of blood and the deformability of erythrocytes can change in different seasons of the animal's wakefulness period to adequately provide tissues with oxygen. The aim of this study was to identify possible adaptive changes in erythrocyte deformability and erythrocyte indices in ground squirrels during their active period. The deformability of erythrocytes was studied by ektacytometry in an osmotic gradient. It was found that in spring, after the arousal of ground squirrels, erythrocytes have the highest deformability (El_max), hydration (O_hyper), water permeability (El_min) and osmotic stability (∆O). In summer, compared with spring, the deformability of erythrocytes decreases, and the average volume of erythrocytes (MCV) also decreases. In autumn, before hibernation, the integral deformability of erythrocytes, their hydration, as well as the range of osmotic stability of erythrocytes increases relative to the summer period. In summer and autumn, in contrast to spring, the average concentration of hemoglobin in the erythrocyte (MCHC) increases. At low shear stress (1 Pa) in summer and autumn, osmoscan becomes a pronounced polymodal form, indicating a change in the viscoelastic properties of ground squirrel erythrocyte membranes. Thus, for the first time, we discovered seasonal variability in the deformability of ground squirrel erythrocytes, which is consistent with the spring-summer activity of animals and preparation for hibernation.

First Data on PAE Levels in Surface Water in Lakes of the Eastern Coast of Baikal.

The increasing consumption of phthalates (PAEs), along with their high toxicity and high mobility, poses a threat to the environment. This study presents initial data on the contents of six priority PAEs in the water of lakes located on the eastern shore of Lake Baikal-Arangatui, Bormashevoe, Dukhovoe, Kotokel, and Shchuchye. The mean total concentrations of the six PAEs in lakes Arangatui and Bormashevoe (low anthropogenic load) were comparable to those in Kotokel (medium anthropogenic load, 17.34 µg/L) but were significantly higher (p < 0.05) than in Dukhovoe and Shchuchye (high anthropogenic load, 10.49 and 2.30 µg/L, respectively). DBP and DEHP were the main PAEs in all samples. The DEHP content in lakes Arangatui and Bormashevoe was quite high, and at some sampling sites it exceeded the MACs established by Russian, U.S. EPA, and WHO regulations. The assessment showed that there is no potential risk to humans associated with the presence of PAEs in drinking water. However, the levels of DEHP, DBP, and DnOP in the water pose a potential threat to sensitive aquatic organisms, as shown by the calculated risk quotients (RQs). It is assumed that the origin of the phthalates in the studied lakes is both anthropogenic and biogenic.

Cassava Starch as an Effective Texture Corrector of Fat-Free Dairy Products Based on Symbiotic Starter Culture.

Cassava starch can be an effective texture corrector for dairy products; however, the presence of a starchy taste in such products is undesirable. A possible solution to this problem can be the use of partially hydrolyzed cassava starch; complex microbial amylase preparations, for instance, Amylosubtilin® or Bacillus licheniformis amylases, can be used as an enzyme. The use of these enzyme preparations in low concentration allows it to obtain cassava starches with increased solubility, which can be easily used in the technology of dairy products. Starch is partially hydrolyzed by amylase preparations and does not significantly affect the chemical composition of the protein part of the Symbilact dairy product. Positive dynamics of the rheological and antioxidant properties of the low-fat dairy product "Symbilact" from the enzyme-modified cassava starch during storage were revealed in the researches. AT starches are more able to correct the structure, especially AT-0.5 and AT-1, but to a less extent increase antioxidant properties. As matter of a fact, BT starches exhibit higher antioxidant potential, but less structure correction.

A new species of Ranunculus (Ranunculaceae) from Western Pamir-Alay, Uzbekistan.

New data on the phylogeny of four rare and endemic species of RanunculusL.sect.Ranunculastrum DC. of western Pamir-Alai, one of which is new to science, have been obtained. Ranunculustojibaevii sp. nov., from the Baysuntau Mountains, Western Hissar Range of Uzbekistan, is described. The new species is closely related to R.botschantzevii Ovcz., R.convexiusculus Kovalevsk. and R.alpigenus Kom., but differs in the blade of the radical leaves, which is rounded-reniform, segments 3-5-dissected, each 2-5-partite with elongated, rounded apical lobes. A phylogenetic analysis, using both the nuclear ribosomal internal transcribed spacer (ITS) and cpDNA (matK, rbcL, trnL-trnF), was informative in placing R.tojibaevii in context with its most closely-related species. Discussion on the geographic distribution, updated identification key, a detailed description, insights about its habitat and illustrations are provided.

Individual iron(iii) glycerolate: synthesis and characterisation.

Iron(ii) and iron(iii) salts of strong acids form iron glycerolates on heating at 180 °C with glycerol in the presence of an equivalent amount of alkali. Individual iron(iii) glycerolate was obtained for the first time. When Fe3O4 magnetic nanoparticles were heated with glycerol, an iron(iii) glycerolate shell was formed on their surface.

Erythrocytes of Little Ground Squirrels Undergo Reversible Oxidative Stress During Arousal From Hibernation.

The hibernation of small mammals is characterized by long torpor bouts alternating with short periods of arousal. During arousal, due to a significant increase in oxygen consumption, tissue perfusion, and the launch of thermogenesis in cells, a large amount of reactive oxygen species (ROS) and nitrogen (RNS) can be formed, which can trigger oxidative stress in cells. To estimate this possibility, we studied the intensity of free-radical processes in the red blood cells (RBCs) of little ground squirrels (LGS; Spermophilus pygmaeus) in the dynamics of arousal from hibernation. We found that in the torpid state, the degree of generation of ROS and RNS (8.3%, p>0.09; 20.7%, p<0.001, respectively), the degree of oxidative modification of membrane lipids and RBC proteins is at a low level (47%, p<0.001; 82.7%, p<0.001, respectively) compared to the summer control. At the same time, the activity of superoxide dismutase (SOD) and catalase (CAT) in RBC is significantly reduced (32.8%, p<0.001; 22.2%, p<0.001, respectively), but not the level of glutathione (GSH). In the torpid state, SOD is activated by exogenous GSH in concentration-dependent manner, which indicates reversible enzyme inhibition. During the arousal of ground squirrels, when the body temperature reaches 25°C, RBCs are exposed oxidative stress. This is confirmed by the maximum increase in the level of uric acid (25.4%, p<0.001) in plasma, a marker of oxidative modification of lipids [thiobarbituric acid reactive substances (TBARS); 82%, p < 0.001] and proteins (carbonyl groups; 499%, p < 0.001) in RBC membranes, as well as the decrease in the level of GSH (19.7%, p < 0.001) in erythrocytes relative to the torpid state and activity of SOD and CAT in erythrocytes to values at the Tb 20°C. After full recovery of body temperature, the level of GSH increases, the ratio of SOD/CAT is restored, which significantly reduces the degree of oxidative damage of lipids and proteins of RBC membranes. Thus, the oxidative stress detected at Tb 25°C was transient and physiologically regulated.

Low-Dose Ammonium Preconditioning Enhances Endurance in Submaximal Physical Exercises.

Preconditioning is often used in medicine to protect organs from ischemic damage and in athletes to enhance the performances. We tested whether low-dose ammonium preconditioning (AMP) could have a beneficial effect on physical exercises (PE). We used Cardiopulmonary Exercise Testing (CPET) on a treadmill to investigate the effects of low-dose AMP on the physical exercise capacity of professional track and field athletes and tested twenty-five athletes. Because of the individual differences between athletes, we performed a preliminary treadmill test (Pre-test) and, according to the results, the athletes were randomly allocated into the AMP and control (placebo, PL) group based on the similarity of the total distance covered on a treadmill. In the AMP group, the covered distance increased (11.3 ± 3.6%, p < 0.02) compared to Pre-test. Similarly, AMP significantly increased O2 uptake volume-VO2 (4.6 ± 2.3%, p < 0.03) and pulmonary CO2 output-VCO2 (8.7 ± 2.8%, p < 0.01). Further, the basic blood parameters (pH, pO2, and lactate) shift was lower despite the greater physical exercise progress in the AMP group compared to Pre-test, whereas in the placebo group there were no differences between Pre-test and Load-test. Importantly, the AMP significantly increased red blood cell count (6.8 ± 2.0%, p < 0.01) and hemoglobin concentration (5.3 ± 1.9%, p < 0.01), which might explain the beneficial effects in physical exercise progress. For the first time, we showed that low-dose AMP had clear beneficial effects on submaximal PE.

Design, synthesis, antibacterial activity and toxicity of novel quaternary ammonium compounds based on pyridoxine and fatty acids.

A diverse series of 43 novel "soft antimicrobials" based on quaternary ammonium pyridoxine derivatives which include six-membered acetals and ketals of pyridoxine bound via cleavable linker moieties (amide, ester) with a fragment of fatty carboxylic acid was designed. Nine compounds exhibited in vitro promising antibacterial activity against Gram-positive and Gram-negative bacterial strains with MIC values comparable with reference antiseptics miramistin, benzalkonium chloride and chlorohexidine. On various clinical isolates, the lead compounds 6i and 12a exhibited antibacterial activity comparable with that of benzalkonium chloride while higher than that of miramistin. Moreover, 6i and 12a were able to kill bacteria embedded into the matrix of mono- and dual species biofilms. The treatment of bacterial cells by either 6i and 12a lead to fast depolarization of the membrane suggesting that the membrane is an apparent molecular target of compounds. 6i and 12a were non mutagenic neither in SOS-chromotest nor in Ames test and non-toxic in vivo at acute oral (LD50 > 2000 mg/kg) and cutaneous administration (LD50 > 2500 mg/kg) on mice. Taken together, our data allow suggesting described active compounds as promising starting point for the new antibacterial agents development.

Is Selfie Behavior Related to Psychological Well-being?

Background: The reasons and consequences of people's activity on social networks have not been sufficiently studied. Most studies have focused on identifying the dangers and risks associated with posting self-portraits on social networks, but it is an open question as to whether such behavior serves to increase people's psychological well-being. Objective: We asked ourselves what are the main motives for publishing selfies and whether online activity contributes to psychological well-being. Design: Our study involved 96 respondents from Moscow, mainly psycho logy students, who provided information about their activity on social networks, and completed questionnaires on their motivation, social support, and psychological well-being. Results: Three main motives for publishing selfies were identified: 1) to increase self-esteem; 2) maintain social contacts; and 3) preserve and exchange information. The higher the ratio of selfies taken to preserve information, the higher was the user's psychological well-being. We found significant differences between the characteristics of those participants with high and low activity, and larger and smaller numbers of "friends" in social networks. For those with high activity, their sense of psychological well-being was largely determined by interaction with others. For these persons, communication (including with virtual contacts) was the important resource of well-being. The other group was less dependent on others, and for them, psychological well-being was not related to their activity on social networks. Conclusion: Our results confirm the connection between the personality traits and characteristics of the respondents' network behavior and their psychological well-being. The study showed that the type of correlation constellation differs between groups of respondents depending upon how much or how little they are oriented toward social support.

Microvesicle Formation Induced by Oxidative Stress in Human Erythrocytes.

Extracellular vesicles (EVs) released by different cell types play an important role in many physiological and pathophysiological processes. In physiological conditions, red blood cell (RBC)-derived EVs compose 4-8% of all circulating EVs, and oxidative stress (OS) as a consequence of different pathophysiological conditions significantly increases the amount of circulated RBC-derived EVs. However, the mechanisms of EV formation are not yet fully defined. To analyze OS-induced EV formation and RBC transformations, we used flow cytometry to evaluate cell esterase activity, caspase-3 activity, and band 3 clustering. Band 3 clustering was additionally analyzed by confocal microscopy. Two original laser diffraction-based approaches were used for the analysis of cell deformability and band 3 activity. Hemoglobin species were characterized spectrophotometrically. We showed that cell viability in tert-Butyl hydroperoxide-induced OS directly correlated with oxidant concentration to cell count ratio, and that RBC-derived EVs contained hemoglobin oxidized to hemichrome (HbChr). OS induced caspase-3 activation and band 3 clustering in cells and EVs. Importantly, we showed that OS-induced EV formation is independent of calcium. The presented data indicated that during OS, RBCs eliminated HbChr by vesiculation in order to sacrifice the cell itself, thereby prolonging lifespan and delaying the untimely clearance of in all other respects healthy RBCs.

Novel Bis-Ammonium Salts of Pyridoxine: Synthesis and Antimicrobial Properties.

A series of 108 novel quaternary bis-ammonium pyridoxine derivatives carrying various substituents at the quaternary nitrogen's and acetal carbon was synthesized. Thirteen compounds exhibited antibacterial and antifungal activity (minimum inhibitory concentration (MIC) 0.25-16 µg/mL) comparable or superior than miramistin, benzalkonium chloride, and chlorhexidine. A strong correlation between the lipophilicity and antibacterial activity was found. The most active compounds had logP values in the range of 1-3, while compounds with logP > 6 and logP < 0 were almost inactive. All active compounds demonstrated cytotoxicity comparable with miramistin and chlorhexidine on HEK-293 cells and were three-fold less toxic when compared to benzalkonium chloride. The antibacterial activity of leading compound 5c12 on biofilm-embedded Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli or Pseudomonas aeruginosa was comparable or even higher than that of the benzalkonium chloride. In vivo 5c12was considerably less toxic (LD50 1705 mg/kg) than benzalkonium chloride, miramistine, and chlorhexidine at oral administration on CD-1 mice. An aqueous solution of 5c12 (0.2%) was shown to be comparable to reference drugs efficiency on the rat's skin model. The molecular target of 5c12 seems to be a cellular membrane as other quaternary ammonium salts. The obtained results make the described quaternary bis-ammonium pyridoxine derivatives promising and lead molecules in the development of the new antiseptics with a broad spectrum of antimicrobial activity.

Antibacterial activity profile of miramistin in in vitro and in vivo models.

BACKGROUND: Miramistin is a widely used antiseptic, disinfectant and preservative, and one of the most popular antimicrobial agents on pharmaceutical market of the Russian Federation (http://www.dsm.ru/en/news/385/). However, there is a lack of reported systematic data on antibacterial efficacy of this agent obtained in accordance with the international standards. AIM: This paper represents a systematic study of antibacterial properties of miramistin. Another objective of this work is to evaluate and compare the exploratory performance of in vitro and in vivo protocols of antiseptics' efficacy testing using miramistin as the reference antiseptic. METHODS: Antibacterial activity of 0.1% and 0.2% aqueous solutions of miramistin against two museum strains of S. aureus (ATCC 209p) and E. coli (CDC F-50) was studied. Three standard in vitro laboratory tests (microdilution test, suspension test, and metal surface test), and one in vivo test (on rat's skin) were used. The study was conducted in accordance with the international regulatory documents. RESULTS: Miramistin showed high bactericidal activity against the studied bacterial pathogens in the standard in vitro tests. Thus, in the microdilution test it showed expressed activity against S. aureus (MIC 8 µg/ml, MBC 16 µg/ml) and E. coli (MIC 32 µg/ml, MBC 128 µg/ml). In the suspension test, miramistin decreased the amount of colony forming units by at least 6 log10 units for S. aureus, and by at least 4.5 log10 units for E. coli. Transition to the metal surface test led to significant decrease of antibacterial activity by 1-3 log10 units as compared to the suspension test. Further dramatic reduction of antiseptic activity (by 3-4 log10 units) was observed in in vivo rat skin test. Addition of a protein contaminant (bovine serum albumin) led to a general decrease in the effectiveness of miramistin against the test pathogens (typically, by 1-2 log10 units). An interesting effect of exposure time-dependent reversal of miramistin's specificity to the studied Gram-positive S. aureus and the Gram-negative E. coli organisms was observed in the metal surface test. CONCLUSIONS: The results of this work provide systematic data on antibacterial efficacy of miramistin. They also underscore the need in relevant in vivo models for evaluation of antiseptics' efficacy. While the existing in vitro methods can be successfully applied at the discovery stages, it is necessary to use more realistic in vivo models at more advanced development stages. The observed selectivity reversal effect should be taken into account when carrying out the antiseptics' efficacy testing and surface disinfection procedures.

Glycomyces buryatensis sp. nov., an actinobacterium isolated from steppe soil.

A novel actinobacterial strain, designated 18T, was isolated from a steppe soil sample collected in Buryatia, Russia and subjected to polyphasic taxonomic characterization. The strain was aerobic and Gram-stain-positive. The 16S rRNA gene sequence of strain 18T exhibited highest similarity to Glycomyces paridis CPCC 204357T (97.2 %). Results of phylogenetic analysis showed that strain 18T formed a distinct branch clearly affiliated to the genus Glycomyces. Whole-cell hydrolysates of the isolate contained meso-diaminopimelic acid as the cell-wall diamino acid. The whole-cell sugar profile was found to contain galactose, glucose, ribose and xylose. MK-10(H4) and MK-11(H4) were the predominant menaquinones. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, five unidentified phospholipids and four unidentified polar lipids. The major fatty acids identified were anteiso-C15 : 0 and iso-C15 : 0. The G+C content of the genomic DNA of strain 18T was 68.0 mol% (draft genome sequence). Based on its phylogenetic, phenotypic and chemotaxonomic features, strain 18T was considered to represent a novel species of the genus Glycomyces, for which the name Glycomyces buryatensis sp. nov. is proposed. The type strain of Glycomyces buryatensis is 18T (=JCM 33362T=CGMCC 4.7610T).

Newly isolated lactic acid bacteria from silage targeting biofilms of foodborne pathogens during milk fermentation.

BACKGROUND: Raw milk, meat and plant materials are subjected to high risks of contamination by various pathogenic bacteria and thus their growth prevention is a great challenge in the food industry. Food fermentation by lactic acid bacteria (LAB) besides changing its organoleptic characteristics also helps to eliminate unfavorable microflora and represses growth of pathogens. To the date only few LABs has been reported to exhibit activity against bacteria embedded in the biofilms characterized by extreme resistance to antimicrobials, high exchange rate with resistance genes and represent high risk factor for foodborne disease development. RESULTS: Six novel LAB strains isolated from the clover silage exhibited pronounced antibacterial activity against biofilm embedded pathogens. We show explicitly that these strains demonstrate high acidification rate, completely repress the growth of E. coli, S. aureus and to a lesser extent P. aeruginosa as well as exhibit appropriate probiotic and milk-fermenting properties. Moreover, in contrast to the approved probiotic strain Lactobacillus plantarum 8PA3, the new isolates were able to efficiently eradicate preformed biofilms of these pathogens and prevent bacterial spreading originating from the biofilm. We suggest these strains as potential additives to the pre-cultures of conventional LAB strains as efficient tools targeting foodborne pathogens in order to prevent food contamination from either seeded raw material or biofilm-fouled equipment. CONCLUSIONS: The AG10 strain identified as L. plantarum demonstrate attractive probiotic and milk fermentation properties as well as high resistance to simulated gastric conditions thus appearing perspective as a starter culture for the prevention of bacterial contamination originating from fouled equipment during milk fermentation.