Non-pathogenic Microorganisms Research Articles

Small RNA discovery in the interaction between barley and the powdery mildew pathogen

BackgroundPlants encounter pathogenic and non-pathogenic microorganisms on a nearly constant basis. Small RNAs such as siRNAs and miRNAs/milRNAs influence pathogen virulence and host defense responses. We exploited the biotrophic interaction between the powdery mildew fungus, Blumeria graminis f. sp. hordei (Bgh), and its diploid host plant, barley (Hordeum vulgare) to explore fungal and plant sRNAs expressed during Bgh infection of barley leaf epidermal cells.ResultsRNA was isolated from four fast-neutron immune-signaling mutants and their progenitor over a time course representing key stages of Bgh infection, including appressorium formation, penetration of epidermal cells, and development of haustorial feeding structures. The Cereal Introduction (CI) 16151 progenitor carries the resistance allele Mla6, while Bgh isolate 5874 harbors the AVRa6 avirulence effector, resulting in an incompatible interaction. Parallel Analysis of RNA Ends (PARE) was used to verify sRNAs with likely transcript targets in both barley and Bgh. Bgh sRNAs are predicted to regulate effectors, metabolic genes, and translation-related genes. Barley sRNAs are predicted to influence the accumulation of transcripts that encode auxin response factors, NAC transcription factors, homeodomain transcription factors, and several splicing factors. We also identified phasing small interfering RNAs (phasiRNAs) in barley that overlap transcripts that encode receptor-like kinases (RLKs) and nucleotide-binding, leucine-rich domain proteins (NLRs).ConclusionsThese data suggest that Bgh sRNAs regulate gene expression in metabolism, translation-related, and pathogen effectors. PARE-validated targets of predicted Bgh milRNAs include both EKA (effectors homologous to AVRk1 and AVRa10) and CSEP (candidate secreted effector protein) families. We also identified barley phasiRNAs and miRNAs in response to Bgh infection. These include phasiRNA loci that overlap with a significant proportion of receptor-like kinases, suggesting an additional sRNA control mechanism may be active in barley leaves as opposed to predominant R-gene phasiRNA overlap in many eudicots. In addition, we identified conserved miRNAs, novel miRNA candidates, and barley genome mapped sRNAs that have PARE validated transcript targets in barley. The miRNA target transcripts are enriched in transcription factors, signaling-related proteins, and photosynthesis-related proteins. Together these results suggest both barley and Bgh control metabolism and infection-related responses via the specific accumulation and targeting of genes via sRNAs.

Detoxification Strategies for Zearalenone Using Microorganisms: A Review.

Zearalenone (ZEA) is a mycotoxin produced by Fusarium fungi that is commonly found in cereal crops. ZEA has an estrogen-like effect which affects the reproductive function of animals. It also damages the liver and kidneys and reduces immune function which leads to cytotoxicity and immunotoxicity. At present, the detoxification of mycotoxins is mainly accomplished using biological methods. Microbial-based methods involve zearalenone conversion or adsorption, but not all transformation products are nontoxic. In this paper, the non-pathogenic microorganisms which have been found to detoxify ZEA in recent years are summarized. Then, two mechanisms by which ZEA can be detoxified (adsorption and biotransformation) are discussed in more detail. The compounds produced by the subsequent degradation of ZEA and the heterogeneous expression of ZEA-degrading enzymes are also analyzed. The development trends in the use of probiotics as a ZEA detoxification strategy are also evaluated. The overall purpose of this paper is to provide a reliable reference strategy for the biological detoxification of ZEA.

An Overview of Ocular Microbiology: Ocular Microbiota, the Effect of Contact Lenses and Ocular Disease

The human eye is an organ that is exposed to the environment continuously, which makes the ocular surface subjected to different types of pathogenic and non-pathogenic microorganisms. Different types of microorganisms can be found naturally in each layer of the eye, such as the Grampositive bacteria (i.e., the Coagulase-negative staphylococci including S. epidermidis and Bacillus sp), the Gram-negative bacteria (as Pseudomonas sp. including P. aeruginosa) and fungi. Apparently, bacteria are considered to be the major causative agent of ocular infections that are believed to be a global problem.

Microorganisms in Biostimulants

In March 2016, the EU Commission presented a proposal for new regulations on fertilising material. The regulation includes product rules for a wide range of organic and inorganic products. Microbial biostimulants is one of the categories of products that are included. Biostimulants, in the draft EU regulation, are defined as fertilising materials that affect nutrient processes independently of the product's own nutrient content and with the purpose of improving nutrient utilisation, tolerance for abiotic stress or quality of the crop. Positive list in which species of these bacterial genera are listed: Azotobacter spp, Rhizobium spp., Azospirillum spp and Mycorrhizal fungi are a part of the regulation.
 Since the import and use of these organisms are the responsibility of both the Norwegian Food Safety Authority and the Norwegian Environment Agency, they asked VKM to submit a joint report on effects on health (humans, plants and animals), biodiversity and dispersal, quality of agricultural land and on soil environment.
 Conclusions:
 Health risks:
 Based upon our literature review, we have found no indication of any specific diseases in plants, animals or humans induced by the discussed microorganisms. A few reported cases of human disease are caused through wound infections or injections in immunocompromised patients. These represent a situation where any microorganism may induce infections and is not specific for the agents discussed in this report. In summary, the risk of any disease caused by the discussed microorganisms is considered negligible.
 Environmental risks:
 In soil the biodiversity, competition, adaptation and functional redundancy of microorganisms are extremely high. This means that introduced microorganisms have a very small chance for establishing, and even less so for affecting biodiversity and soil functioning. Introduction of nitrogen fixing species or fungi that can transport P to plants (mycorrhiza) will lead to an increase in the primary production. However, even a large increased activity for these processes will not outcompete naturally occurring symbiotic N-fixation or growth of inherently non-mycorrhizal plant species. Thus, the risks associated with introduced non-pathogenic microorganisms are very low.

S-Layer Glycoprotein From Lactobacillus kefiri Exerts Its Immunostimulatory Activity Through Glycan Recognition by Mincle.

The development of new subunit vaccines has promoted the rational design of adjuvants able to induce a strong T-cell activation by targeting specific immune receptors. The S-layer is a (glyco)-proteinaceous envelope constituted by subunits that self-assemble to form a two-dimensional lattice that covers the surface of different species of Bacteria and Archaea. Due to their ability to self-assemble in solution, they are attractive tools to be used as antigen/hapten carriers or adjuvants. Recently, we have demonstrated that S-layer glycoprotein from Lactobacillus kefiri CIDCA 8348 (SLP-8348) enhanced the LPS-induced response on macrophages in a Ca2+-dependent manner, but the receptors involved in these immunomodulatory properties remain unknown. Therefore, we aim to determine the C-type lectin receptors (CLRs) recognizing this bacterial surface glycoprotein as well as to investigate the role of glycans in both the immunogenicity and adjuvant capacity of SLP-8348. Here, using a mild periodate oxidation protocol, we showed that loss of SLP-8348 glycan integrity impairs the cell-mediated immune response against the protein. Moreover, our data indicate that the adjuvant capacity of SLP-8348 is also dependent of the biological activity of the SLP-8348 glycans. In order to evaluate the CLRs involved in the interaction with SLP-8348 an ELISA-based method using CLR–hFc fusion proteins showed that SLP-8348 interacts with different CLRs such as Mincle, SingR3, and hDC-SIGN. Using BMDCs derived from CLR-deficient mice, we show that SLP-8348 uptake is dependent of Mincle. Furthermore, we demonstrate that the SLP-8348-induced activation of BMDCs as well as its adjuvant capacity relies on the presence of Mincle and its signaling adaptor CARD9 on BMDCs, since SLP-8348-activated BMDCs from Mincle−/− or CARD9−/− mice were not capable to enhance OVA-specific response in CD4+ T cells purified from OT-II mice. These findings significantly contribute to the understanding of the role of glycans in the immunomodulation elicited by bacterial SLPs and generate a great opportunity in the search for new adjuvants derived from non-pathogenic microorganisms.

Validation of microbiological testing in cardiovascular tissue establishments: results of a second international quality-round trial

The performance of many laboratories can be evaluated by participation in external quality assessment (EQA) schemes. EQA allows for comparison of a laboratory's performance with a source outside the laboratory-either a peer group of laboratories or a reference laboratory. Such EQA schemes do not exist in tissue banking despite the fact that tissue establishments (TE) perform very complex procedures. This paper describes the first ever EQA scheme in the field specifically assessing microbiological aspects in heart valve (HV) banking. Twenty-two TEs participated. Three HV tissue samples were sent to each participating TE-two contaminated with non-pathogenic micro-organisms and a third negative control. The aims were to isolate and identify the micro-organisms present and then to successfully decontaminate the HV tissue using the routine standard operating procedures of the TE. Eight of the TEs were able to isolate and identify all contaminating micro-organisms present, and of these, five also successfully decontaminated the tissue; 13 TEs failed to establish the identity of one or more of the contaminants; five TEs appear to have introduced contamination during the processing or testing of the tissue; and eight failed to successfully decontaminate the HV tissue. This initiative provides TEs with an international benchmark for tissue product microbiology testing. It has identified significant variation in practice and in the ability of different TEs to identify the presence of contamination. There is now work ongoing with the aim of setting up a regular EQA scheme for HV banking.

Probiotics as Beneficial Dietary Supplements to Prevent and Treat Cardiovascular Diseases: Uncovering Their Impact on Oxidative Stress.

The gut microbiota, the ecosystem formed by a wide symbiotic community of nonpathogenic microorganisms that are present in the distal part of the human gut, plays a prominent role in the normal physiology of the organism. The gut microbiota's imbalance, gut dysbiosis, is directly related to the origin of various processes of acute or chronic dysfunction in the host. Therefore, the ability to intervene in the gut microbiota is now emerging as a possible tactic for therapeutic intervention in various diseases. From this perspective, evidence is growing that a functional dietary intervention with probiotics, which maintain or restore beneficial bacteria of the digestive tract, represents a promising therapeutic strategy for interventions in cardiovascular diseases and also reduces the risk of their occurrence. In the present work, we review the importance of maintaining the balance of the intestinal microbiota to prevent or combat such processes as arterial hypertension or endothelial dysfunction, which underlie many cardiovascular disorders. We also review how the consumption of probiotics can improve autonomic control of cardiovascular function and provide beneficial effects in patients with heart failure. Among the known effects of probiotics is their ability to decrease the generation of reactive oxygen species and, therefore, reduce oxidative stress. Therefore, in this review, we specifically focus on this antioxidant capacity and its relationship with the beneficial cardiovascular effects described for probiotics.

Host feeding behaviour of Dermacentor reticulatus males in relation to the transmission of pathogens.

The three-host Dermacentor reticulatus tick transmits many pathogens, which are introduced into the host with saliva during feeding. The aim of present study was to analyse the behaviour of males of this species on the host in homogeneous sex groups comprising 15 (group I) and 30 (group II) male specimens and in a mixed group composed of 15 males and 15 females (group III). Although the dynamics of attachment of the males to host skin slightly differed between these three groups, there was no statistically significant difference in the duration of the attachment process. The duration of host attachment of the males in groups I, II, and III at 18±2°C and 50 ± 2% RH was 2.43 ± 2.46, 7.75 ± 11.85, and 9.07 ± 10.97 days, respectively. After 7-9-day feeding, the male engorgement weight (MEW) was similar, regardless of the size of the group and the presence or absence of females on the host. Tick males ingest a small amount of host blood. The value of MEW in the three groups did not differ statistically from the weight of unengorged specimens. The host feeding behaviour of D. reticulatus males and their effective feeding suggest that they may play a role in transmission of pathogens and non-pathogenic microorganisms with tick saliva during blood meal ingestion. Clinical and laboratory diagnostics of human and animal tick-borne diseases as well as epidemiological studies should consider the possibility of pathogen transmission by males of metastriata ticks, which feed on the host for several days.

In vitro assessment of the probiotic potential of lactobacilli isolated from Minas artisanal cheese produced in the Araxá region, Minas Gerais state, Brazil

ABSTRACT Minas artisanal cheese is made from endogenous starter cultures, including lactic acid bacteria (LAB). Some LAB may possess probiotic potential. Thus, this study aimed to evaluate the in vitro probiotic properties of lactobacilli isolated from Minas artisanal cheeses produced in Minas Gerais. Ten samples of lactobacilli, formerly isolated from those cheeses, were submitted to the following assays: antimicrobial susceptibility, tolerance to artificial gastric juice and biliary salts, production of hydrogen peroxide and antagonism against pathogenic and non-pathogenic micro-organisms. Only L. plantarum (C0) was sensitive to all tested antimicrobials, while the other LAB samples were resistant to at least one drug. Six samples were tolerant to artificial gastric juice, and L. brevis (A6) even grew in that medium. Three samples were tolerant to biliary salts. Only L. brevis (E35) produced hydrogen peroxide. Difference (P< 0.05) was observed among the means of inhibition haloes of lactobacilli against Enterococcus faecalis ATCC 19433 and Lactobacillus plantarum C24 in spot-on-the-lawn assay. All samples of lactobacilli inhibited Escherichia coli ATCC 25922, Salmonella enterica var. Typhimurium ATCC 14028 in co-culture antagonism test (P< 0.0001). Most lactobacilli samples showed in vitro probiotic potential. From the tested samples, L. brevis (A6) presented the best results considering all in vitro probiotic tests.

Biocleaning to Remove Graffiti: A Real Possibility? Advances towards a Complete Protocol of Action

The first academic studies on the use of microorganisms in cleaning procedures appeared in the late 1980s/early 1990s. In the past thirty years, most of such studies have addressed the removal of nitrate and sulphate salts and organic matter from surfaces by using non-pathogenic anaerobic microorganisms, mainly sulphate-reducing bacteria. The successful use of microbes in the removal of graffiti paint remains, however, a work in progress. Biocleaning surfaces to remove graffiti is not a simple task, because of the complex chemical composition of graffiti paints. This study looks at ways of improving the bioremoval of graffiti and presents the latest findings regarding different methodological aspects of cleaning natural and man-made stone. Granite and concrete substrates were coated with silver and black graffiti spray paints for comparison of the efficacy of the biocleaning method on these different materials. Visual and microscopic examination along with colour and infrared measurements made after application of the bacterial strains tested (previously shown to be suitable candidates for bioremoval of graffiti) revealed remarkably successful results. The findings presented thus represent progress in the development of a biocleaning protocol applicable to the in-situ removal of graffiti. Important improvements have been made regarding the time of treatment, which has been reduced by up to 20 days, and the use of a culture medium enriched with powdered graffiti, which facilitates and accelerates the adaptation of the microorganisms to the target surface.

Pathogenic and Non-Pathogenic Microorganisms in the Rapid Alert System for Food and Feed.

The most frequently notified pathogenic microorganisms in the RASFF in 1980–2017 were Salmonella sp., Listeria, Escherichia and Vibrio, whereas, among the notified non-pathogenic microorganisms were unspecified microorganisms, Enterobacteriaceae, Salmonella sp. and Coliforms. Microorganisms were reported mainly in poultry meat, meat, fish, molluscs, crustaceans, fruits, vegetables, herbs, spices, nuts, milk, cereals (in food) and in feed materials and pet food (in feed). The number of notifications decreased at the turn of 2005 and 2006, but has steadily increased since then. The notification basis were official controls, border controls and company’s checks. Products were notified mainly by Italy, France, United Kingdom, Germany and Netherlands. The reported products originated from Brazil, European Union countries and India, Thailand and Vietnam. The notification types were alerts, information and border rejections. The distribution status was often not specified or distribution on the market was possible. The risk decision was usually not made. Products were re-dispatched, import was not authorised or products were withdrawn from the market, destroyed and recalled from the market. Proper cooperation within the framework of the RASFF can contribute to shaping public health law and reducing outbreaks associated with microorganisms.

A cross-sectional screening by next-generation sequencing reveals Rickettsia, Coxiella, Francisella, Borrelia, Babesia, Theileria and Hemolivia species in ticks from Anatolia

BackgroundTicks participate as arthropod vectors in the transmission of pathogenic microorganisms to humans. Several tick-borne infections have reemerged, along with newly described agents of unexplored pathogenicity. In an attempt to expand current information on tick-associated bacteria and protozoans, we performed a cross-sectional screening of ticks, using next-generation sequencing. Ticks seeking hosts and infesting domestic animals were collected in four provinces across the Aegean, Mediterranean and Central Anatolia regions of Turkey and analyzed by commonly used procedures and platforms.ResultsTwo hundred and eighty ticks comprising 10 species were evaluated in 40 pools. Contigs from tick-associated microorganisms were detected in 22 (55%) questing and 4 feeding (10%) tick pools, with multiple microorganisms identified in 12 pools. Rickettsia 16S ribosomal RNA gene, gltA, sca1 and ompA sequences were present in 7 pools (17.5%), comprising feeding Haemaphysalis parva and questing/hunting Rhipicephalus bursa, Rhipicephalus sanguineus (sensu lato) and Hyalomma marginatum specimens. A near-complete genome and conjugative plasmid of a Rickettsia hoogstraalii strain could be characterized in questing Ha. parva. Coxiella-like endosymbionts were identified in pools of questing (12/40) as well as feeding (4/40) ticks of the genera Rhipicephalus, Haemaphysalis and Hyalomma. Francisella-like endosymbionts were also detected in 22.5% (9/40) of the pools that comprise hunting Hyalomma ticks in 8 pools. Coxiella-like and Francisella-like endosymbionts formed phylogenetically distinct clusters associated with their tick hosts. Borrelia turcica was characterized in 5% (2/40) of the pools, comprising hunting Hyalomma aegyptium ticks. Co-infection of Coxiella-like endosymbiont and Babesia was noted in a questing R. sanguineus (s.l.) specimen. Furthermore, protozoan 18S rRNA gene sequences were detected in 4 pools of questing/hunting ticks (10%) and identified as Babesia ovis, Hemolivia mauritanica, Babesia and Theileria spp.ConclusionsOur metagenomic approach enabled identification of diverse pathogenic and non-pathogenic microorganisms in questing and feeding ticks in Anatolia.

Isolation and Characterization of Endophytic Bacteria from Piper longum

Endophytes are non-pathogenic microorganisms that reside within internal tissues of plant, without causing any apparent symptoms of infection. In an attempt to identify endophytes from Piper longum, the Indian long pepper, a combination of conventional and molecular approaches was used. Using culture-dependent approach, six different bacterial isolates were obtained from various surface-sterilized parts (roots, nodes, internodes, petioles, leaves and spikes) of the plant. In general, roots harboured maximum concentration of endophytic bacterial isolates, while leaves contained the minimum levels. These endophytes were analyzed on the basis of colony morphology and biochemical characteristics. Based on the results obtained after BLAST search of 16S rDNA sequence in NCBI database, the endophytes isolated from Piper longum showed highest similarity to Endophytic bacteria 135L-3, Enterobacter sp. SQ6-43, Bacillus casamancensis strain TN3, Alishewanella sp. JS-30, Bacterium B28 and Enterobacter ludwigii strain g45. Most of the identified bacteria belong to the phylum Proteobacteria and Firmicutes, whose members have been reported to act as growth-promoting bacteria in other plant species. Some of these endophytes tested positive for the ability to produce indole-3-acetic acid and hydrogen cyanide, suggesting their potential roles in plant growth promotion and biological control against pathogens, respectively.

ИСПОЛЬЗОВАНИЕ ПОЧВООБИТАЮЩИХ БАКТЕРИЙ ДЛЯ ЗАЩИТЫ САХАРНОЙ СВЕКЛЫ ОТ КОРНЕВЫХ ГНИЛЕЙ

In the recent years, the use of soil microorganisms has generated great interests in crop production by reducing the damage of diseases. Actualite.The application of PGPR bacteria makes possible to improve the quantity and quality of plant s harvest and to reduce chemical pressure on the environment. The aim of investigations - study of the ability of the soil saprophytic diazotrophs bacteria (PGPR) reducing the infectious background of root rot diseases. deposited in the National Collection of nonpathogenic microorganisms of Moldova - Peudomonas aureofaciens CNMN-PsB-05, Pseudomonas fluorescens CNMN-PsB-04, Pseudomonas putida-PsB-06, Bacillus subtilis CNMN-bB-08. Materials. The agents pathogens from the beet tuber were isolated on potato-glucose agar and were identification as Fusarium oxysporum and Fusarium gibbosum, They reduce the shoots of beet seeds for 60-75%. The ability for antagonism strains Peudomonas aureofaciens CNMN-PsB-05, Pseudomonas fluorescensCNMN-PsB-04, Pseudomonas putida - PsB- 06, Bacillus subtilis CNmN-BB-08, deposited in the National Collection of nonpathogenic microorganisms of Moldova was defined in laboratory tests. Results. The suspension of bacterial strains with titre 108 CFU/ml show the antagonism in vitro toward the fitopathogens in time cultivation combine in Petri plates: the radius of delay growth Fusarium spp. were establish. Traitment of beet seeds, inoculate with pathogens, in time sowing with bacterials suspensions decreased the level of infection from agents pathogens of root rot, increased the shoots of beet seeds for 20-30%, the weight of tuber in 2.2 4,0 one and improved production of plants.

Fruit Stem-End Rot

After harvest, the fruit ripens and stem-end rot (SER) starts to develop, leading to significant fruit losses. SER is caused by diverse pathogenic fungi that endophytically colonize the stem during fruit development in the orchard or field and remain quiescent until the onset of fruit ripening. During the endophytic-like stage, the pathogenic fungus colonizes the phloem and xylem of the fruit stem-end; after fruit ripening, the fungus converts to a necrotrophic lifestyle, while colonizing the fruit parenchyma, and causes SER. The fruit stem-end is colonized not only by pathogenic fungi, but also by various nonpathogenic endophytic microorganisms, including fungi, yeast and bacteria. However, little is known about the fruit stem-end endophytic microbiome, which could contain new and existing biocontrol agents. To control fruit SER, treatments such as ripening inhibition, harvesting with the stem, application of chemical or biological fungicides, or physical control such as heat treatments, cold storage, or exposure to light have been suggested. This review focuses on the characterization of SER pathogens, the stem-end microbiome, and different pre- and postharvest practices that could control fruit SER.

Antimicrobial Efficacy and Chemical Properties of Caryophyllus aromaticus and Origanum majorana Essential Oils Against Foodborne Bacteria Alone and in Combination

Background: Food products need to be protected against pathogenic and non-pathogenic microorganisms. One method is adding antimicrobial agents. Consumers’ tendency to use synthetic additives is drastically decreasing due to their side effects and also the emergence of multidrug resistant microorganisms. Plant essential oils (EOs) are natural antimicrobial compounds which are widely used in food industry. Objective: The objectives were to determine the chemical compositions of Caryophyllus aromaticus and Origanum majorana EOs and also to assess their antimicrobial activities against foodborne bacteria alone and in combination. Materials and Methods: The EOs were analyzed by gas chromatography-mass spectrometry. Antibacterial activities of the EOs against foodborne bacteria were assessed using disc diffusion method. The minimum inhibitory concentration (MIC) values of the EOs were determined by microdilution broth method and then minimum bactericidal concentration (MBC) values were determined. Checkerboard synergy testing was performed to determine the fractional inhibitory concentration index. Then time-kill curves were drawn based on the bacterial population (CFU/mL) against time (h). Results: The major constituents of C. aromaticus were eugenol and carvacrol, while O. majorana had carvacrol, thymol, trans-caryophyllene, and cymene as the main constituents. Zone of inhibition for O. majorana EO was greater than that for C. aromaticus EO. The inhibition zone of O. majorana EO against all the tested bacteria except for Bacillus subtilis was significantly greater than that of streptomycin (P ˂ 0.05). MIC value of the EOs against bacteria was 0.1% except for O. majorana EO against B. subtilis (0.3%). MBC values of C. aromaticus and O. majorana EOs ranged from 0.5% to 1.0% and 0.3% to 0.5% (v/v), respectively. The EOs were more effective on gram-positive bacteria than gram-negative ones. The combination of EOs revealed synergistic activity against Listeria monocytogenes, partial synergistic activity against B. subtilis, and additive effect against Salmonella typhimurium, Escherichia coli O157:H7 and Staphylococcus aureus were indifferent against the combination of EOs. Time–kill curves of the EOs demonstrated strong bactericidal effect against all foodborne bacteria at 6 and 24 hours either alone or in combination. Conclusion: The synergistic, partial synergistic, and additive effects of the combination of C. aromaticus and O. majorana EOs strengthen the antimicrobial activity, expand the spectrum of activity, reduce the concentrations required, decrease the side effects, and prevent the alteration of organoleptic properties of food.

Implications of the expression of Enterococcus faecalis citrate fermentation genes during infection.

Citrate is an ubiquitous compound in nature. However, citrate fermentation is present only in a few pathogenic or nonpathogenic microorganisms. The citrate fermentation pathway includes a citrate transporter, a citrate lyase complex, an oxaloacetate decarboxylase and a regulatory system. Enterococcus faecalis is commonly present in the gastro-intestinal microbiota of warm-blooded animals and insect guts. These bacteria can also cause infection and disease in immunocompromised individuals. In the present study, we performed whole genome analysis in Enterococcus strains finding that the complete citrate pathway is present in all of the E. faecalis strains isolated from such diverse habitats as animals, hospitals, water, milk, plants, insects, cheese, etc. These results indicate the importance of this metabolic preservation for persistence and growth of E. faecalis in different niches. We also analyzed the role of citrate metabolism in the E. faecalis pathogenicity. We found that an E. faecalis citrate fermentation-deficient strain was less pathogenic for Galleria mellonella larvae than the wild type. Furthermore, strains with deletions in the oxaloacetate decarboxylase subunits or in the α-acetolactate synthase resulted also less virulent than the wild type strain. We also observed that citrate promoters are induced in blood, urine and also in the hemolymph of G. mellonella. In addition, we showed that citrate fermentation allows E. faecalis to grow better in blood, urine and G. mellonella. The results presented here clearly indicate that citrate fermentation plays an important role in E. faecalis opportunistic pathogenic behavior.

Mycoplasmas and Novel HO-1 Inducers: Recent Advances.

The development of an effective system for the treatment of inflammatory diseases requires comprehensive studies of the cellular signaling molecular networks comprising responses to various stressors, including pathogenic and non-pathogenic microorganisms. Significant attention on fundamental and applied research has recently focused on inducers of hemе oxygenase-1 (HO-1) and inhibitors of the expression of this enzyme, which regulates expression of this and other cytoprotective molecules and modulation of inflammation. Recent studies indicate that mycoplasmas (a major group of human pathogens of the Mollicutes) are capable of modulating inflammatory responses through the activation of the Nrf2 and the expression of HO-1. In vitro experiments demonstrate that the membrane lipoproteins (LAMPs), along with lipoprotein derivatives (lipopeptide MALP-2) in mycoplasmas cause a "cross-talk" between the pro- and antiinflammatory signaling pathways. Importantly, lipopeptide/lipoprotein - induced expression of HO-1 tends to suppress inflammation. The study of the molecular network that causes the corresponding outcome can facilitate the development of new approaches for the treatment of inflammatory processes. The derivatives of LAMPs and MALP-2 and of their analogues may prove promising for the treatment of diseases associated with chronic inflammation.

Innovations in the Therapy of Atopic Dermatitis Complicated by a Secondary Infection

Reduced skin barrier properties in patients with atopic dermatitis (AtD) are largely caused by microbiome changes and extensive Staphylococcus aureus colonisation of the skin. In this regard, the integument of patients with AtD requires constant care and the use of various emollients. The inclusion of lysates of non-pathogenic microorganisms and prebiotics in the composition of emollients ensures the normalisation of the microbiome composition and the immunological barrier of the skin. The article presents the results of our own observations on the application of two cosmetic scin-care products for damaged skin with vitamin F in children with AtD complicated by a secondary infection, while the composition of one of the products is additionally enriched with ceramides and prebiotics. The safety and high efficacy of both products have been shown, however, the presence of ceramides and prebiotics in the emollient composition makes it possible to achieve a marked decrease in the degree of S. aureus colonisation of the skin.

Application of pulsed electric fields for the elimination of highly drug-resistant Candida grown under modelled microgravity conditions

AbstractCandida lusitaniaeandC. guilliermondiiare perfect model organisms for the study ofCandidagenera behaviour in various conditions. Both of them are rare pathogens capable to cause candidiasis in the patients with weakened immune system and can undergo morphology switches related to the increased antifungal drug resistance.Candidagenera yeasts are able to inhabit diverse range of ecological niches including space ships and space stations. During the long-term expeditions, astronauts are affected by various factors that can change the state immune system. In such conditions, the commensal usually non-pathogenic microorganisms can spread through the body of the host and cause infections. Weakened immune system and limited use of drugs in spaceships promote the search of the alternative methods for the biocontrol of microorganisms. Several studies demonstrate that microorganisms are altering their gene expression, physiology, morphology, pathogenicity and evolving resistance to the antifungals under microgravity conditions. Our research indicated that switch to the pseudohyphae morphology leads up 30-fold increased resistance to amphotericin B inC. lusitaniaeandC. guilliermondii. Cultivation of yeasts in rotary cell culture system (RCCS) is related to the altered cell growth and resistance to the antifungal treatment. Our results showed that growth in the RCCS led to the extreme increase in cell resistance to amphotericin B as compared with the standard growth conditions. In our research, we applied electroporation for the biocontrol of twoCandidaspecies.C. lusitaniaeandC. guilliermondiicells grown in RCCS exhibited significantly increased survivability after pulsed electric field (PEF) treatment in comparison with cells grown under routine conditions. We have shown that PEF bursts of 2.5–25 kV cm−1of 100 µs × 8 duration display a dose-dependent permeabilization of both studiedCandidaspecies. Our research indicated that budding cells and pseudohyphae morphology cells, with increased resistance to amphotericin B, can be effectively inactivated after applying PEF higher than 15 kV cm−1.