Adhesion of Klebsiella oxytoca to bladder or lung epithelial cells is promoted by the presence of other opportunistic pathogens.

The intestinal and respiratory tracts of healthy individuals serve as habitats for a diverse array of microorganisms, among which Klebsiella oxytoca holds significance as a causative agent in numerous community- and hospital-acquired infections, often manifesting in polymicrobial contexts. In specific circumstances, K. oxytoca, alongside other constituents of the gut microbiota, undergoes translocation to distinct physiological niches. In these new environments, it engages in close interactions with other microbial community members. As this interaction may progress to co-infection where the virulence of involved pathogens may be promoted and enhance disease severity, we investigated how K. oxytoca affects the adhesion of commonly co-isolated bacteria and vice versa during co-incubation of different biotic and abiotic surfaces. Co-incubation was beneficial for the adhesion of at least one of the two co-cultured strains. K. oxytoca enhanced the adhesion of other enterobacteria strains to polystyrene and adhered more efficiently to bladder or lung epithelial cell lines in the presence of most enterobacteria strains and S. aureus. This effect was accompanied by bacterial coaggregation mediated by carbohydrate-protein interactions occurring between bacteria. These interactions occur only in sessile, but not planktonic populations, and depend on the features of the surface. The data are of particular importance for the risk assessment of the urinary and respiratory tract infections caused by K. oxytoca, including those device-associated. In this paper, we present the first report on K. oxytoca ability to acquire increased adhesive capacities on epithelial cells through interactions with common causal agents of urinary and respiratory tract infections.

Inorganic polyphosphate and ion transport across biological membranes.

Inorganic polyphosphate (polyP) is widely recognized for playing important roles and processes involved in energy and phosphate storage, regulation of gene expression, and calcium signaling. The less well-known role of polyP is as a direct mediator of ion transport across biological membranes. Here, we will briefly summarize current knowledge of the molecular mechanisms of how polyP can be involved in membrane ion transport. We discuss three types of mechanisms that might involve polyP: (1) formation of non-protein channel complex that includes calcium, polyP, and polyhydroxybutyrate (PHB); (2) modulation of the channel activity of PHBlated protein channels; and (3) direct effects of polyP on the function of the voltage-gated ion channels in the process that do not involve PHB.

Induced Expression of CYP51a and HK1 Genes Associated with Penconazole and Fludioxonil Resistance in the Potato Pathogen Fusarium oxysporum.

Preventing antifungal resistance development and identifying pathogens with high, medium, and low risk of resistance development to a particular fungicide or fungicide class is crucial in the fight against phytopathogens. We characterized the sensitivity of potato wilt-associated Fusarium oxysporum isolates to fludioxonil and penconazole and assessed the effect of these fungicides on the expression of fungal sterol-14-α-demethylase (CYP51a) and histidine kinase (HK1) genes. Penconazole stunted the growth of F. oxysporum strains at all concentrations used. While all isolates were susceptible to this fungicide, concentrations of up to 1.0 µg/mL were insufficient to cause a 50% inhibition. At low concentrations (0.63 and 1.25 µg/mL), fludioxonil stimulated growth in F. oxysporum. With an increase in the concentration of fludioxonil, only one strain (F. oxysporum S95) exhibited moderate sensitivity to the fungicide. Interaction of F. oxysporum with penconazole and fludioxonil leads to respective elevated expressions of the CYP51a and HK1 genes, which upsurge with increasing concentration of the fungicides. The data obtained indicate that fludioxonil may no longer be suitable for potato protection and its continuous use could only lead to an increased resistance with time.

Biofilm Formation by Mutant Strains of Bacilli under Different Stress Conditions.

Bacillus subtilis is traditionally classified as a PGPR that colonizes plant roots through biofilm formation. The current study focused on investigating the influence of various factors on bacilli biofilm formation. In the course of the study, the levels of biofilm formation by the model strain B. subtilis WT 168 and on its basis created regulatory mutants, as well as strains of bacilli with deleted extracellular proteases under conditions of changes in temperature, pH, salt and oxidative stress and presence of divalent metals ions. B. subtilis 168 forms halotolerant and oxidative stress-resistant biofilms at a temperature range of 22 °C-45 °C and a pH range of 6-8.5. The presence of Ca2+, Mn2+ and Mg2+ upsurges the biofilm development while an inhibition with Zn2+. Biofilm formation level was higher in protease-deficient strains. Relative to the wild-type strain, degU mutants showed a decrease in biofilm formation, abrB mutants formed biofilms more efficiently. spo0A mutants showed a plummeted film formation for the first 36 h, followed by a surge after. The effect of metal ions and NaCl on the mutant biofilms formation is described. Confocal microscopy indicated that B. subtilis mutants and protease-deficient strains differ in matrix structure. The highest content of amyloid-like proteins in mutant biofilms was registered for degU-mutants and protease-deficient strains.

Toolkit for cellular studies of mammalian mitochondrial inorganic polyphosphate.

Introduction: Inorganic polyphosphate (polyP) is an ancient polymer which is extremely well-conserved throughout evolution, and found in every studied organism. PolyP is composed of orthophosphates linked together by high-energy bonds, similar to those found in ATP. The metabolism and the functions of polyP in prokaryotes and simple eukaryotes are well understood. However, little is known about its physiological roles in mammalian cells, mostly due to its unknown metabolism and lack of systematic methods and effective models for the study of polyP in these organisms. Methods: Here, we present a comprehensive set of genetically modified cellular models to study mammalian polyP. Specifically, we focus our studies on mitochondrial polyP, as previous studies have shown the potent regulatory role of mammalian polyP in the organelle, including bioenergetics, via mechanisms that are not yet fully understood. Results: Using SH-SY5Y cells, our results show that the enzymatic depletion of mitochondrial polyP affects the expression of genes involved in the maintenance of mitochondrial physiology, as well as the structure of the organelle. Furthermore, this depletion has deleterious effects on mitochondrial respiration, an effect that is dependent on the length of polyP. Our results also show that the depletion of mammalian polyP in other subcellular locations induces significant changes in gene expression and bioenergetics; as well as that SH-SY5Y cells are not viable when the amount and/or the length of polyP are increased in mitochondria. Discussion: Our findings expand on the crucial role of polyP in mammalian mitochondrial physiology and place our cell lines as a valid model to increase our knowledge of both mammalian polyP and mitochondrial physiology.

Biotechnological Key Genes of the Rhodococcus erythropolis MGMM8 Genome: Genes for Bioremediation, Antibiotics, Plant Protection, and Growth Stimulation.

Anthropogenic pollution, including residues from the green revolution initially aimed at addressing food security and healthcare, has paradoxically exacerbated environmental challenges. The transition towards comprehensive green biotechnology and bioremediation, achieved with lower financial investment, hinges on microbial biotechnology, with the Rhodococcus genus emerging as a promising contender. The significance of fully annotating genome sequences lies in comprehending strain constituents, devising experimental protocols, and strategically deploying these strains to address pertinent issues using pivotal genes. This study revolves around Rhodococcus erythropolis MGMM8, an associate of winter wheat plants in the rhizosphere. Through the annotation of its chromosomal genome and subsequent comparison with other strains, its potential applications were explored. Using the antiSMASH server, 19 gene clusters were predicted, encompassing genes responsible for antibiotics and siderophores. Antibiotic resistance evaluation via the Comprehensive Antibiotic Resistance Database (CARD) identified five genes (vanW, vanY, RbpA, iri, and folC) that were parallel to strain CCM2595. Leveraging the NCBI Prokaryotic Genome Annotation Pipeline (PGAP) for biodegradation, heavy metal resistance, and remediation genes, the presence of chlorimuron-ethyl, formaldehyde, benzene-desulfurization degradation genes, and heavy metal-related genes (ACR3, arsC, corA, DsbA, modA, and recG) in MGMM8 was confirmed. Furthermore, quorum-quenching signal genes, critical for curbing biofilm formation and virulence elicited by quorum-sensing in pathogens, were also discerned within MGMM8's genome. In light of these predictions, the novel isolate MGMM8 warrants phenotypic assessment to gauge its potential in biocontrol and bioremediation. This evaluation extends to isolating active compounds for potential antimicrobial activities against pathogenic microorganisms. The comprehensive genome annotation process has facilitated the genetic characterization of MGMM8 and has solidified its potential as a biotechnological strain to address global anthropogenic predicaments.

Identification, characterization, and genome sequencing of Brevibacterium sediminis MG-1 isolate with growth-promoting properties.

In recent years, plant growth-promoting rhizobacteria (PGPR) have received increased attention due to their prospective use as biofertilizers for the enhancement of crop growth and yields. However, there is a growing need to identify new PGPR isolates with additional beneficial properties. In this paper, we describe the identification of a new strain of a non-sporulating Gram-positive bacterium isolated from the rhizosphere of potato plants, classified as Brevibacterium sediminis MG-1 based on whole-genome sequencing. The bacteria are aerobic; they grow in a pH range of 6.0-10.0 (optimum 6.0), and a temperature range of 20-37 °C (optimum 30 °C). At 96 h of cultivation, strain MG-1 synthesizes 28.65 µg/ml of indole-3-acetic acid (IAA) when 500 µg/ml of l-tryptophan is added. It is a producer of catechol-type siderophores and ACC deaminase (213 ± 12.34 ng/ml) and shows halotolerance. Treatment of pea, rye, and wheat seeds with a suspension of MG-1 strain cells resulted in the stimulation of stem and root biomass accumulation by 12-26% and 6-25% (P < 0.05), respectively. Treatment of seeds with bacteria in the presence of high salt concentration reduced the negative effects of salt stress on plant growth by 18-50%. The hypothetical gene lin, encoding the bacteriocin Linocin-M18, RIPP-like proteins, and polyketide synthase type III (T3PKS) loci, gene clusters responsible for iron acquisition and metabolism of siderophores, as well as gene clusters responsible for auxin biosynthesis, were identified in the B. sediminis MG-1 genome. Thus, the rhizosphere-associated strain B. sediminis MG-1 has growth-stimulating properties and can be useful for the treatment of plants grown on soils with high salinity. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03392-z.

Are formae speciales pathogens really host specific? A broadened host specificity in Fusarium oxysporum f.sp. radicis-cucumerinum.

Phytopathogenic strains of Fusarium oxysporum Schlecht exhibit clear host specificity, which appears to be a persistent characteristic and a dependable base for the forma specialis system of these pathogens. Here, we report an altered host specificity of the F. oxysporum f.sp. radicis-cucumerinum strain V03-2 g (Forc V03-2 g) - a causative agent of cucumber root-rot, the clonal derivates of which acquired the ability to infect tomato plants. Since the clonal derivates of Forc V03-2 g with transformed host specificity preserved their ability to parasitize on cucumber plants, the changes that occurred can be classified as broadening of host specificity. To our knowledge, this is the first observation of pathogenicity changes in formae speciales of F. oxysporum. The clonal derivates acquired could be used to trace genetic determinants of the host specificity of phytopathogenic strains of F. oxysporum.

Diversity in the swimming motility and flagellar regulon structure of uropathogenic Morganella morganii strains / Diversidad en la motilidad natatoria y la estructura del regulón flagelar de cepas uropatógenas de Morganella morganii

In current times, the opportunistic pathogen Morganella morganii is increasingly becoming a cause of urinary tract infections. The condition has been further complicated by the multiple drug resistance of most isolates. Swimming motility plays an important role in the development of urinary tract infections, allowing bacteria to colonize the upper urinary tract. We determined the differences between the growth, swimming motility, and biofilm formation of two M. morganii strains MM 1 and MM 190 isolated from the urine of patients who had community-acquired urinary tract infections. MM 190 showed a lower growth rate but better-formed biofilms in comparison to MM 1. In addition, MM 190 possessed autoaggregation abilities. It was found that a high temperature (37 °C) inhibits the flagellation of strains and makes MM 190 less motile. At the same time, the MM 1 strain maintained its rate of motility at this temperature. We demonstrated that urea at a concentration of 1.5% suppresses the growth and swimming motility of both strains. Genome analysis showed that MM 1 has a 17.7-kb-long insertion in flagellar regulon between fliE and glycosyltransferase genes, which was not identified in corresponding loci of MM 190 and 9 other M. morganii strains with whole genomes. Both strains carry two genes encoding flagellin, which may indicate flagellar antigen phase variation. However, the fliC2 genes have only 91% identity to each other and exhibit some variability in the regulatory region. We assume that all these differences influence the swimming motility of the strains.(AU)

Diversity in the swimming motility and flagellar regulon structure of uropathogenic Morganella morganii strains.

In current times, the opportunistic pathogen Morganella morganii is increasingly becoming a cause of urinary tract infections. The condition has been further complicated by the multiple drug resistance of most isolates. Swimming motility plays an important role in the development of urinary tract infections, allowing bacteria to colonize the upper urinary tract. We determined the differences between the growth, swimming motility, and biofilm formation of two M. morganii strains MM 1 and MM 190 isolated from the urine of patients who had community-acquired urinary tract infections. MM 190 showed a lower growth rate but better-formed biofilms in comparison to MM 1. In addition, MM 190 possessed autoaggregation abilities. It was found that a high temperature (37 °C) inhibits the flagellation of strains and makes MM 190 less motile. At the same time, the MM 1 strain maintained its rate of motility at this temperature. We demonstrated that urea at a concentration of 1.5% suppresses the growth and swimming motility of both strains. Genome analysis showed that MM 1 has a 17.7-kb-long insertion in flagellar regulon between fliE and glycosyltransferase genes, which was not identified in corresponding loci of MM 190 and 9 other M. morganii strains with whole genomes. Both strains carry two genes encoding flagellin, which may indicate flagellar antigen phase variation. However, the fliC2 genes have only 91% identity to each other and exhibit some variability in the regulatory region. We assume that all these differences influence the swimming motility of the strains.

Supplementation of Bacillus subtilis GM5 enhances broiler body weight gain and modulates cecal microbiota.

We investigated the effect of the strain Bacillus subtilis GM5 on growth, feed conversion, and the composition of cecum microbiota in broiler chickens. Half of which received a control diet, while the other half was fed a diet supplemented with GM5 spores. Cecal contents on days 1, 10, and 42 were subjected to metataxonomic analysis. Principal Component Analysis showed that the control and probiotic groups formed three separate clusters, indicating changes, which occurred gradually in microbial communities. On day 1, Firmicutes (53.87-57.61%) and Proteobacteria (43.77-38.93%) were prevalent in both groups, whereas samples of days 10 and 42 were predominantly occupied by Firmicutes (54.55-81.79%) and Bacteroidetes (26.94-30.45%). In the group of chickens treated with probiotic, the average daily gain in body weight was higher, while feed conversion decreased by 1.44%. A surge in the presence of beneficial bacteria of the Ruminococcaceae family was observed. The introduction of the probiotic led to an elevated Firmicutes/Bacteroidetes ratio, which positively correlated with chickens' bodyweight (Spearman ρ = 1.0, P < 0.05). Supplementing broiler feed with B. subtilis GM5 spores leads to improved feed intake and digestibility, which is paramount in reducing the cost of the final product. Thus, the probiotic strain GM5 modulates the cecal microbiota of broiler chickens and increases microbial diversity, which is well exhibited on the 42nd day. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02634-2.

Structure and variation of root-associated microbiomes of potato grown in alfisol.

Root-associated fungi and bacteria play a pivotal role in the plant-soil ecosystem by influencing both plant growth and immunity. The aim of this study was to unravel the biodiversity of the bacterial and fungal rhizosphere (RS) and rhizoplane (RP) microbiota of Zhukovskij rannij potato (Solanum tuberosum L.) cultivar growing in the Alfisol of Tatarstan, Russia. To assess the structure and diversity of microbial communities, we employed the 16S rRNA and internal transcribed spacer gene library technique. Overall, sequence analysis showed the presence of 3982 bacterial and 188 fungal operational taxonomic units (OTUs) in the RP, and 6018 bacterial and 320 fungal OTUs for in the RS. Comparison between microbial community structures in the RS and RP showed significant differences between these compartments. Biodiversity was higher in the RS than in the RP. Although members of Proteobacteria (RS-59.1 ± 4.9%; RP-54.5 ± 9.2%), Bacteroidetes (RS-23.19 ± 10.2%; RP-34.52 ± 10.4%) and Actinobacteria (RS-11.55 ± 4.9%; RP-7.7 ± 5.1%) were the three most dominant phyla, accounting for 94-98% of all bacterial taxa in both compartments, notable variations were observed in the primary dominance of classes and genera in RS and RP samples. In addition, our results demonstrated that the potato rhizoplane was significantly enriched with the genera Flavobacterium, Pseudomonas, Acinetobacter and other potentially beneficial bacteria. The fungal community was predominantly inhabited by members of the Ascomycota phylum (RS-81.4 ± 8.1%; RP-81.7 ± 5.7%), among which the genera Fusarium (RS-10.34 ± 3.41%; RP-9.96 ± 4.79%), Monographella (RS-7.66 ± 4.43%; RP-9.91 ± 5.87%), Verticillium (RS-4.6 ± 1.43%; RP-8.27 ± 3.63%) and Chaetomium (RS-4.95 ± 2.07%; RP-8.33 ± 4.93%) were particularly abundant. Interestingly, potato rhizoplane was significantly enriched with potentially useful fungal genera, such as Mortierella and Metacordiceps. A comparative analysis revealed that the abundance of Fusarium (a cosmopolitan plant pathogen) varied significantly depending on rotation variants, indicating a possible control of phytopathogenic fungi via management-induced shifts through crop rotational methods. Analysis of the core microbiome of bacterial and fungal community structure showed that the formation of bacterial microbiota in the rhizosphere and rhizoplane is dependent on the host plant.

"We'll check vital signs only till we finish the school": experiences of student nurses regarding intra-semester clinical placement in Ghana.

BACKGROUND: Clinical practicum is an integral part of nursing education because it provides students with opportunities to perform nursing care and practice specific nursing tasks. In Ghana, little is known about the experiences of baccalaureate student nurses with regard to intra-semester clinical practicum. This study therefore, explored perceptions, challenges, and how the intra-semester clinical practicum affects the learning process of student nurses in a private university in Ghana. METHODS: Exploratory descriptive phenomenological design was used. Nine in-depth interviews and three focus group discussions were conducted for baccalaureate student nurses in their second, third and fourth years of study. Only those who have attended intra-semester clinical practicum for at least two semesters in the course of their study were recruited. Purposive sampling technique was used to select the participants. The sample size was based on data saturation, however, a total of 33 participants were recruited. Data was analysed using content analysis technique. RESULTS: The findings show that baccalaureate student nurses perceive the intra-semester clinical practicum as beneficial. It affords the opportunity to translate theoretical knowledge into practice concurrently. However, students recounted their stressful experiences during the clinical period which negatively affected their academic work. Additionally, staff nurses assigned the students to do menial jobs instead of appropriate nursing tasks. CONCLUSIONS: A review of the "block" method in which students will go to clinicals for a stipulated number of consecutive days in a month and then resume lectures, is worth considering.