Decreased Biofilm Formation Research Articles

Microplastics colonized by Hafnia paralvei through biofilm formation regulated by c-di-GMP and cAMP promote its spread in water

Microplastics (MPs) colonized by pathogens pose significant risks to the environment and health of animals and humans, however, the strategies for pathogens colonization in MPs and the effects of its colonization on spread of pathogens have not been fully characterized. Here, we investigated the biofilm formation mechanism regulated by c-di-GMP in Hafnia paralvei Z11, and determined the effect of MPs colonized by H. paralvei Z11 on the spread of strain Z11. Overexpression of yhjH, a c-di-GMP phosphodiesterase gene, attenuated intracellular c-di-GMP level in strain Z11, leading to an increase in biofilm dispersal and a decrease in biofilm formation. Meanwhile, the decline of c-di-GMP inhibited the expression of cAMP phosphodiesterase genes, increasing the cAMP content and promoting bacterial motility, that was responsible for the increase of biofilm dispersal. Furthermore, the formation of biofilms by strain Z11 on MPs promotes its colonization, which contributes to its vertical and horizontal spread in water after colonizing polyvinyl chloride-MPs and polypropylene-MPs, respectively. Therefore, this study reveals, for the first time, MPs colonized by H. paralvei Z11 through biofilms regulated by crosstalk between c-di GMP and cAMP promote the spread of strain Z11 in water, which expands the understanding of colonization strategy of pathogens on MPs and its risk on spread of pathogens.

Characterizing the interplay between Acinetobacter baumannii, A549 cells, and anti-Omp34 antibodies: implications for adherence, internalization, and cytotoxicity.

Acinetobacter baumannii thrives within eukaryotic cells, influencing persistence, treatment approaches, and progression of disease. We probed epithelial cell invasion by A. baumannii and the influence of antibodies raised to outer membrane protein 34 (Omp34) on epithelial interactions. We expressed and purified recombinant Omp34 and induced anti-Omp34 antibodies in Bagg albino or BALB/c mice. Omp34 was evaluated for acute toxicity in mice through histological analysis of six organs. The host cell line, A549, was exposed to both A. baumannii 19606 and a clinical isolate. The study also investigated serum resistance, adherence, internalization, and proliferation of A. baumannii in A549 cells, with and without anti-Omp34 sera, utilizing cell culture techniques and light microscopy. A549 cell viability was evaluated by A. baumannii challenge and exposure to anti-Omp34 sera. Actin disruption experiments using cytochalasin D probed microfilament and microtubule roles in A. baumannii invasion. Omp34 prompted antibody production without toxicity in mice. The serum showed bactericidal effects on both strains. Additionally, both A. baumannii strains were found to form biofilms. Omp34 serum was observed to decrease biofilm formation, bacterial adherence, internalization, and proliferation in A549 cells. Furthermore, the use of anti-Omp34 serum enhanced the post-infection survival of the host cell. Pre-exposure of A549 cells to cytochalasin D reduced bacterial internalization, highlighting the role of actin polymerization in the invasion process. Microscopic analysis revealed various interactions, such as adherence, membrane alterations, vacuolization, apoptosis, and cellular damage. Anti-Omp34 serum-exposed A549 cells were protected and showed reduced damage. The findings reveal that A. baumannii can significantly multiply intracellularly within host cells. This suggests the bacterium's ability to establish an environment conducive to its replication by preventing fusion with degradative lysosomes and inhibiting acidification. This finding contributes to the understanding of A. baumannii's intracellular persistence and highlights the role of Omp34 in influencing apoptosis, autophagy, and bacterial adherence, which may impact the development of effective treatments against A. baumannii infections.

Effect of Acetic Acid on Biofilm Formation in Paracidovorax citrulli, Causal Agent of Bacterial Fruit Blotch.

The unique tissue structure of pathogenic bacteria biofilm plays an important role in its pathogenicity and bactericide resistance. Inhibition or destruction of biofilm formation of pathogenic bacteria is of great significance for the control of plant bacterial diseases. In this study, Paracidovorax citrulli was inoculated into KB medium containing acetic acid, and after shaking at 28°C and 55 r/min for 48 h, it was found that the content of extracellular polysaccharide, extracellular protein and extracellular DNA (eDNA) decreased with the increase of acetic acid concentration, which resulted in the decrease of biofilm formation, it is not even possible to form biofilms on plastic slides. When the final concentration of acetic acid in the culture medium was greater than or equal to 0.5 mg/mL, there was no biofilm on the plastic slides. Therefore, the use of acetic acid as an inhibitor of P. citrulli has a good potential for control of bacterial fruit blotch.

The Influence of Streptococcus mutans Biofilm Formation in a Polymicrobial Environment (Streptococcus gordonii & Porphyromonas gingivalis).

Biofilms play a vital role in the occurrence or worsening of an infectious disease. Streptococcus mutans is a bacterium with the ability to form biofilms that plays a key role in the development of infectious diseases such as dental caries. The formation of biofilms in S. mutans is mediated by quorum sensing. Inhibiting quorum sensing can be considered as one of the approaches to prevent caries. This study aims to investigate the ability of Streptococcus gordonii and Porphyromonas gingivalis bacteria to inhibit the formation of S. mutans biofilm. This research was conducted to analyze bacterial biofilm formation and metabolism. The bacteria used are S. mutans (serotype C), S. gordonii (ATCC 5165), and P. gingivalis (ATCC 33277). Biofilm formation was analyzed using the crystal violet assay. Bacterial metabolism was analyzed using the methylthiazol tetrazolium (MTT) assay. The results of the crystal violet assay indicate a decrease in biofilm formation in S. mutans when in the presence of S. gordonii and S. mutans in the presence of P. gingivalis. The results of the MTT assay show no significant change in the bacterial metabolism of S. mutans in the presence of S. gordonii and S. mutans in the presence of P. gingivalis. However, S. mutans with the presence of S. gordonii and P. gingivalis show an increase in biofilm formation and bacterial metabolism. S. gordonii and P. gingivalis are each capable of inhibiting the formation of S. mutans biofilm in a polymicrobial environment.

Nutmeg vs mace: Evaluation of functional, nutritional, antioxidant and antimicrobial traits to employ as a food preservative

Nutmeg and mace from Myristica fragrans Houtt. are renowned for their culinary significance and medicinal properties. This study was designed to analyze the functional properties, proximate and mineral composition of nutmeg and mace, phytochemical estimation, antioxidant and antimicrobial potential of nutmeg and mace essential oils and oleoresins. The study was further elaborated by testing their synergistic interaction with streptomycin and antibiofilm potential of the most effective treatment. Results revealed better functional properties, higher fiber (35.2%) and mineral (K, Mg, P, Ca, Mn, Zn) content in nutmeg than mace. Nutmeg and mace essential oils were isolated by hydrodistillation and their respective oleoresins were prepared by the Soxhlet extraction method. Phytochemical screening showed a greater phenolic content in mace oleoresin (43.85 mg GAE/g of dw) and terpenoid content in mace essential oil (75.4%). Among all the treatments, mace oleoresin revealed best antioxidant potential against mustard oil degradation during its accelerated oxidation at 70 °C when stored for 7–28 days with minimum increase in peroxide, anisidine, thiobarbituric acid and total oxidation values of mustard oil. Mace oleoresin also revealed the maximum antibacterial activity against Staphylococcus aureus, Bacillus spp., Escherichia coli and Yersinia enterocolitica (MIC values; 180–530 μg/ml). It also revealed synergistic interaction with streptomycin against all the test bacteria along with a notable decrease in biofilm formation (50% of MIC.) GC-MS studies of mace oleoresin showed the presence of terpenoids, phenylpropanoids, phenols and fatty acids and their esters. Hence, mace oleoresins may be used as a natural and safe food preservative.

Bacteriophage P2-71: a promising therapeutic against multidrug-resistant Proteus mirabilis in urinary tract infections.

Proteus mirabilis is a Gram-negative, rod-shaped bacterium widely found in natural environments. It is known for causing a range of severe illnesses in mammals, particularly urinary tract infections (UTIs). This study evaluates the therapeutic efficacy of phage P2-71 against Proteus mirabilis in vivo and in vitro environments. The in vitro therapeutic potential of bacteriophage P2-71 was assessed through the ability of phage to kill Proteus mirabilis by using a plate counting assay, and biofilm inhibition and biofilm lysis assays using a microtitre plate method. Additionally, an in vivo UTI model in C57BL/6Jmice was developed via urethral inoculation of the bacterium. Phage therapy was administered through urethral injection over a period of 5 days. Therapeutic outcomes were measured by analyzing bacterial load, phage titer, inflammatory markers, and histopathological changes in the urine, urogenital tissues, and spleen. In vitro, bacteriophage P2-71 achieved significant reductions in P. mirabilis concentrations, with log reductions of 1.537 and 0.7009 CFU/mL in laboratory and urine environments, respectively (p < 0.001). The phage also decreased biofilm formation by 34-49% and lysed 15-25% of mature biofilms at various multiplicities of infection (MOIs) (p < 0.001). In vivo, phage treatment significantly lowered bacterial concentrations in the urine on Days 1 and 3 (p < 0.0001), achieving a maximum reduction of 4.602 log₁₀ CFU/mL; however, its effectiveness diminished by Day 5 (p > 0.05). Concurrently, phage titers decreased over time. Importantly, phage treatment notably reduced bacterial load in the bladder, kidneys, and spleen (p < 0.001). Inflammatory markers such as IL-6, IL-1β, and TNF-α were significantly lower in the treatment group, especially in the bladder (p < 0.0001), indicating an effective reduction in inflammation. Histopathological analysis showed significant mitigation of tissue damage. The results demonstrated that bacteriophage P2-71 is a promising alternative therapy for UTIs caused by MDR Proteus mirabilis. This bacteriophage therapy offers a viable strategy for managing infections where traditional antimicrobials fail, highlighting its potential in clinical applications.

Antimicrobial Functionalization of Surfaces by a Chimeric Adhesive Protein.

The main aim of this work is to account for the prevention and control of microbial growth on surfaces of interest in medical technology. Surface modification is often achieved by physiotherapy or chemical treatments that can involve time-consuming steps, hazardous reagents, and harsh conditions. One of the ways to overcome these drawbacks is the use of surface-active proteins such as hydrophobins. They can form stable protein layers on different surfaces, serving as anchoring points for other molecules of interest. The fungal hydrophobin Vmh2, already exploited for its adhesive ability, has been fused with the antimicrobial peptide GKY20, forming the chimeric protein used herein for functionalizing polystyrene (PS) and bacterial cellulose (BC). As a natural biomass, BC has multiple advantages, including biodegradability, low cost, renewability, high purity, and excellent mechanical properties. The chimeric protein has been proven to successfully adhere to both surfaces. A strong decrease in biofilm formation on PS and a good bactericidal effect of BC have been demonstrated. These findings provide evidence of an alternative strategy to obtain functional composites using a green, easy process.

The Effect of the Lysine Acetylation Modification of ClpP on the Virulence of Vibrio alginolyticus.

Acetylation modification has become one of the most popular topics in protein post-translational modification (PTM) research and plays an important role in bacterial virulence. A previous study indicated that the virulence-associated caseinolytic protease proteolytic subunit (ClpP) is acetylated at the K165 site in Vibrio alginolyticus strain HY9901, but its regulation regarding the virulence of V. alginolyticus is still unknown. We further confirmed that ClpP undergoes lysine acetylation (Kace) modification by immunoprecipitation and Western blot analysis and constructed the complementation strain (C-clpP) and site-directed mutagenesis strains including K165Q and K165R. The K165R strain significantly increased biofilm formation at 36 h of incubation, and K165Q significantly decreased biofilm formation at 24 h of incubation. However, the acetylation modification of ClpP did not affect the extracellular protease (ECPase) activity. In addition, we found that the virulence of K165Q was significantly reduced in zebrafish by in vivo injection. To further study the effect of lysine acetylation on the pathogenicity of V. alginolyticus, GS cells were infected with four strains, namely HY9901, C-clpP, K165Q and K165R. This indicated that the effect of the K165Q strain on cytotoxicity was significantly reduced compared with the wild-type strain, while K165R showed similar levels to the wild-type strain. In summary, the results of this study indicate that the Kace of ClpP is involved in the regulation of the virulence of V. alginolyticus.

Effect of intraspecies interactions on Pseudomonas aeruginosa biofilm formation and gene expression

Pseudomonas aeruginosa is a common pathogen of acute and chronic infections that can increase risk to burned patients and those with chronic wounds, especially when forming biofilms. These complex ecosystems are formed by several bacteria that communicate, cooperate, and compete with each other. To enhance its survival capacity in such a competitive environment, P. aeruginosa produces secondary metabolites with a range of biological activities, among other factors. Here, we evaluated how such bioactive molecules mediate intraspecies interactions to modulate P. aeruginosa biofilm formation. An overall decrease in biofilm formation on the surface of stainless steel coupons was observed for both dual-strain and single-strain biofilms formed in the presence of secondary metabolites extracted from other P. aeruginosa strains. Modulation of biofilm formation by these metabolites during P. aeruginosa intraspecies interactions suggests a role for these molecules in strain competition. The most likely mechanism by which it occurs might be modulation of iron uptake. Among the genes analyzed, those related to import (tonB1) and biosynthesis (pvdS) of the siderophore pyoverdine were the most downregulated during exposure to the metabolite extracts. Exploring P. aeruginosa intraspecies interactions during biofilm formation might be an interesting approach to understand its lifestyle better and, thus, the mechanisms involved in the species’ biofilm formation.

Different Roles of Dioxin-Catabolic Plasmids in Growth, Biofilm Formation, and Metabolism of Rhodococcus sp. Strain p52.

Microorganisms harbor catabolic plasmids to tackle refractory organic pollutants, which is crucial for bioremediation and ecosystem health. Understanding the impacts of plasmids on hosts provides insights into the behavior and adaptation of degrading bacteria in the environment. Here, we examined alterations in the physiological properties and gene expression profiles of Rhodococcus sp. strain p52 after losing two conjugative dioxin-catabolic megaplasmids (pDF01 and pDF02). The growth of strain p52 accelerated after pDF01 loss, while it decelerated after pDF02 loss. During dibenzofuran degradation, the expression levels of dibenzofuran catabolic genes on pDF01 were higher compared to those on pDF02; accordingly, pDF01 loss markedly slowed dibenzofuran degradation. It was suggested that pDF01 is more beneficial to strain p52 under dibenzofuran exposure. Moreover, plasmid loss decreased biofilm formation, especially after pDF02 loss. Transcriptome profiling revealed different pathways enriched in upregulated and downregulated genes after pDF01 and pDF02 loss, indicating different adaptation mechanisms. Based on the transcriptional activity variation, pDF01 played roles in transcription and anabolic processes, while pDF02 profoundly influenced energy production and cellular defense. This study enhances our knowledge of the impacts of degradative plasmids on native hosts and the adaptation mechanisms of hosts, contributing to the application of plasmid-mediated bioremediation in contaminated environments.

Lactoferrin Affects the Viability of Bacteria in a Biofilm and the Formation of a New Biofilm Cycle of Mannheimia haemolytica A2.

Respiratory diseases in ruminants are responsible for enormous economic losses for the dairy and meat industry. The main causative bacterial agent of pneumonia in ovine is Mannheimia haemolytica A2. Due to the impact of this disease, the effect of the antimicrobial protein, bovine lactoferrin (bLf), against virulence factors of this bacterium has been studied. However, its effect on biofilm formation has not been reported. In this work, we evaluated the effect on different stages of the biofilm. Our results reveal a decrease in biofilm formation when bacteria were pre-incubated with bLf. However, when bLf was added at the start of biofilm formation and on mature biofilm, an increase was observed, which was visualized by greater bacterial aggregation and secretion of biofilm matrix components. Additionally, through SDS-PAGE, a remarkable band of ~80 kDa was observed when bLf was added to biofilms. Therefore, the presence of bLf on the biofilm was determined through the Western blot and Microscopy techniques. Finally, by using Live/Dead staining, we observed that most of the bacteria in a biofilm with bLf were not viable. In addition, bLf affects the formation of a new biofilm cycle. In conclusion, bLf binds to the biofilm of M. haemolytica A2 and affects the viability of bacteria and the formation a new biofilm cycle.

Diverse polysaccharide production and biofilm formation abilities of clinical Klebsiella pneumoniae.

Klebsiella pneumoniae infections have become a growing threat for human health. The lack of understanding of the relationship between antibiotic resistance, mucoviscosity, and biofilm formation in clinical isolates impedes our abilities to effectively predict K. pneumoniae infection outcomes. These traits are also associated with fitness in natural populations and more specifically within a host. The Multidrug-Resistant Organism Repository and Surveillance Network offers a unique opportunity into the genetic and phenotypic variabilities in the K. pneumoniae isolates encountered in the clinics today. To this end, we compared the genetic profiles of these isolates with the phenotypic biofilm formation abilities, percent mucoviscosity, and growth rates. We found most isolates formed limited biofilm, although a select group of isolates could form extremely robust biofilms. Variation in biofilm formation could not be explained by difference in growth rate, suggesting specific genetic and physical determinants. Interestingly, the most mucoid strains in the populations were lacking the genetic element regulating the mucoid phenotype and three of these isolates were able to form robust biofilms. There was a significant phenotype-genotype correlation with decreased biofilm formation and an insertion sequence in the transcriptional activator of the type III fimbrial system. Finally, confocal microscopy highlighted the structural and spatial heterogeneity of biofilm among the most robust biofilm formers not detected by traditional methods. The combination of phenotypic, genomic and image analyses allowed us to reveal an unexpected phenotypic diversity and an intricate relation between growth, mucoviscosity and specific virulence-associated genetic determinants.

Genomic and induction evidence for bacteriophage contributions to sargassum-bacteria symbioses

BackgroundSymbioses between primary producers and bacteria are crucial for nutrient exchange that fosters host growth and niche adaptation. Yet, how viruses that infect bacteria (phages) influence these bacteria-eukaryote interactions is still largely unknown. Here, we investigate the role of viruses on the genomic diversity and functional adaptations of bacteria associated with pelagic sargassum. This brown alga has dramatically increased its distribution range in the Atlantic in the past decade and is predicted to continue expanding, imposing severe impacts on coastal ecosystems, economies, and human health.ResultsWe reconstructed 73 bacterial and 3963 viral metagenome-assembled genomes (bMAGs and vMAGs, respectively) from coastal Sargassum natans VIII and surrounding seawater. S. natans VIII bMAGs were enriched in prophages compared to seawater (28% and 0.02%, respectively). Rhodobacterales and Synechococcus bMAGs, abundant members of the S. natans VIII microbiome, were shared between the algae and seawater but were associated with distinct phages in each environment. Genes related to biofilm formation and quorum sensing were enriched in S. natans VIII phages, indicating their potential to influence algal association in their bacterial hosts. In-vitro assays with a bacterial community harvested from sargassum surface biofilms and depleted of free viruses demonstrated that these bacteria are protected from lytic infection by seawater viruses but contain intact and inducible prophages. These bacteria form thicker biofilms when growing on sargassum-supplemented seawater compared to seawater controls, and phage induction using mitomycin C was associated with a significant decrease in biofilm formation. The induced metagenomes were enriched in genomic sequences classified as temperate viruses compared to uninduced controls.ConclusionsOur data shows that prophages contribute to the flexible genomes of S. natans VIII-associated bacteria. These prophages encode genes with symbiotic functions, and their induction decreases biofilm formation, an essential capacity for flexible symbioses between bacteria and the alga. These results indicate that prophage acquisition and induction contribute to genomic and functional diversification during sargassum-bacteria symbioses, with potential implications for algae growth.1vVC6YGWLCm_DZc419o4RCVideo

MutL Significantly Regulates the Formation of Biofilms in Bacillus amyloliquefaciens YT1

The purpose of this study is to discover and excavate more key factors and signaling pathways that regulate the formation intensity of biofilms and to fully reveal the possible models affecting biofilm formation. By using gene homologous recombination and bioinformatics technology, a MutL protein-directed deletion mutant strain was successfully constructed. The growth status of the mutant strain was observed, and it was confirmed that, except for the change in cell morphology, there were no significant differences in growth and reproduction between the mutant strain and the wild-type strain. By using the induced biofilm formation technique, the significant decrease in biofilm formation in the MutL mutant strain was successfully verified. The plate confrontation test confirmed that the inhibitory ability of the mutant strain against rice blast fungus was not significantly different from that of the WT strain. The colonization ability of the mutant strain on rice stems was tested, and it was confirmed that the colonization ability of the mutant strain was significantly lower than that of the WT strain. In terms of the prevention and control effect of rice blast disease, the mutant strain showed a significant decrease. By using transcriptomic big data, the gene and pathway expression differences between the mutant strain and the WT strain during biofilm formation were analyzed. The analysis revealed no significant correlation with the previously reported spo0A and tapA-sipW-tasA pathways. The key factor capB of the polyglutamic acid signaling pathway, which affects the formation of the biological model, was found to have a significant decrease in expression. A mechanical hypothesis was proposed: MutL may participate in regulating the formation intensity of Bacillus biofilms by regulating the formation of glutamic acid to polyglutamic acid.

Novel trans-Resveratrol Derivatives: Design, Synthesis, Antibacterial Activity, and Mechanisms.

Rice bacterial leaf blight and rice bacterial leaf streak have induced tremendous damage to production of rice worldwide. To discover an effective novel antibacterial agent, a series of novel trans-resveratrol (RSV) derivatives containing 1,3,4-oxadiazole and amide moieties were designed and synthesized for the first time. Most of them showed excellent antibacterial activities against Xanthomonas oryzae pv oryzicola and Xanthomonas oryzae pv oryzae. Especially, compound J12 had the best inhibitory with the half-maximal effective concentration values of 4.2 and 5.0 mg/L, respectively, which were better than that of RSV (63.7 and 75.4 mg/L), bismerthiazol (79.5 and 89.6 mg/L), and thiodiazole copper (105.4 and 112.8 mg/L). Furthermore, compound J12 had an excellent control effect against rice bacterial leaf streak and rice bacterial leaf blight, with protective activities of 46.2 and 42.1% and curative activities of 44.5 and 41.7%, respectively. Preliminary mechanisms indicated that compound J12 could not only remarkably decrease biofilm formation, extracellular polysaccharide production, and the synthesis of extracellular enzymes but also destroy bacterial cell surface morphology, thereby reducing the pathogenicity of bacteria. In addition, compound J12 could increase the activity of defense-related enzymes and affect the expression of multiple pathogenic-related genes including plant-pathogen interaction, the MAPK signaling pathway, and phenylpropanoid biosynthesis, and this could improve the defense of rice against rice bacterial leaf streak infection. The present work indicates that the RSV derivatives can be used as promising candidates for the development of antibacterial agents.

Synthetic Peptides Capable of Potent Multigroup Staphylococcal Quorum Sensing Activation and Inhibition in Both Cultures and Biofilm Communities.

The pathogen Staphylococcus epidermidis uses a chemical signaling process, i.e., quorum sensing (QS), to form robust biofilms and cause human infection. Many questions remain about QS in S. epidermidis, as it uses this intercellular communication pathway to both negatively and positively regulate virulence traits. Herein, we report synthetic multigroup agonists and antagonists of the S. epidermidis accessory gene regulator (agr) QS system capable of potent superactivation and complete inhibition, respectively. These macrocyclic peptides maintain full efficacy across the three major agr specificity groups, and their activity can be "mode-switched" from agonist to antagonist via subtle residue-specific structural changes. We describe the design and synthesis of these non-native peptides and demonstrate that they can appreciably decrease biofilm formation on abiotic surfaces, underscoring the potential for agr agonism as a route to block S. epidermidis virulence. Additionally, we show that both the S. epidermidis agonists and antagonists are active in S. aureus, another common pathogen with a related agr system, yet only as antagonists. This result not only revealed one of the most potent agr inhibitors known in S. aureus but also highlighted differences in the mechanisms of agr agonism and antagonism between these related bacteria. Finally, our investigations reveal unexpected inhibitory behavior for certain S. epidermidis agr agonists at sub-activating concentrations, an observation that can be leveraged for the design of future probes with enhanced potencies. Together, these peptides provide a powerful tool set to interrogate the role of QS in S. epidermidis infections and in Staphylococcal pathogenicity in general.

The Ugd, a capsular polysaccharide synthesis protein, regulates the bacterial motility in Vibrio alginolyticus

Vibrio alginolyticus is one of the most common opportunistic pathogens in marine animals and humans. In this study, A transposon mutation library of the V. alginolyticus E110 was used to identify motility-related genes, and we found three flagellar and one capsular polysaccharide (CPS) synthesis-related genes were linked to swarming motility. Then, gene deletion and complementation further confirmed that CPS synthesis-related gene ugd is involved in the swarming motility of V. alginolyticus. Phenotype assays showed that the Δugd mutant reduced CPS production, decreased biofilm formation, impaired swimming ability, and increased cytotoxicity compared to the wild-type strain. Transcriptome analysis showed that 655 genes (15%) were upregulated and 914 genes (21%) were downregulated in the Δugd strain. KEGG pathway and heatmap analysis revealed that genes involved in two-component systems (TCSs), chemotaxis, and flagella assembly pathways were downregulated in the Δugd mutant. On the other hand, genes involved in pathways of human diseases, biosynthesis ABC transporters, and metabolism were upregulated in the Δugd mutant. The RT-qPCR further validated that ugd-regulated genes are associated with motility, biofilm formation, virulence, and TCSs. These findings imply that ugd may be an important player in the control of some physiological processes in V. alginolyticus, highlighting its potential as a target for future research and potential therapeutic interventions.

Inhibition of some multidrug-resistant bacteria using prepared essential oil Nanoemulsion formulas and their mode of action

Inhibition of some multidrug-resistant bacteria using prepared essential oil Nanoemulsion formulas and their mode of action

The bacillithiol pathway is required for biofilm formation in Staphylococcus aureus

Staphylococcus aureus is a major human pathogen that can cause infections that range from superficial skin and mucosal infections to life threatening disseminated infections. S. aureus can attach to medical devices and host tissues and form biofilms that allow the bacteria to evade the host immune system and provide protection from antimicrobial agents. To counter host-generated oxidative and nitrosative stress mechanisms that are part of the normal host responses to invading pathogens, S. aureus utilizes low molecular weight (LMW) thiols, such as bacillithiol (BSH). Additionally, S. aureus synthesizes its own nitric oxide (NO), which combined with its downstream metabolites may also protect the bacteria against specific host responses. We have previously shown that LMW thiols are required for biofilm formation in Mycobacterium smegmatis and Pseudomonas aeruginosa. Here, we show that the S. aureus bshC mutant strain, which is defective in the last step of the BSH pathway and lacks BSH, is impaired in biofilm formation. We also identify a possible S-nitrosobacillithiol reductase (BSNOR), similar in sequence to an S-nitrosomycothiol reductase found in M. smegmatis and show that the putative S. aureus bsnoR mutant strain has reduced levels of BSH and decreased biofilm formation. Our studies also show that NO plays an important role in biofilm formation and that acidified sodium nitrite severely reduces biofilm thickness. These studies provide insight into the roles of oxidative and nitrosative stress mechanisms on biofilm formation and indicate that BSH and NO are key players in normal biofilm formation in S. aureus.

Elongation Factor P is Required for Processes Associated with Acinetobacter Pathogenesis

Antibiotic resistance is one of the world’s fastest-growing and most prevalent problems today. The influx of antibiotics within our environment from inadequate antibiotic stewardship has led to a surge of drug-resistant microorganisms. The CDC has classified Carbapenem-resistant Acinetobacter (CRA) as an urgent threat within this crisis. New drug development is imperative to combat infections caused by drug-resistant pathogens such as CRA. Bacterial translation, the process of protein synthesis by the ribosome, is a common target for new antibiotic development. Elongation factor P (EF-P) is a universally conserved translation factor required for antibiotic resistance in many bacteria. In this study, we assessed the importance of EF-P in processes associated with Acinetobacter pathogenesis. In the absence of EF-P, Acinetobacter baylyi displays decreased biofilm formation, surface-associated motility, and resistance to beta-lactams and carbapenems. This data holds hope for future drug development targeting EF-P in pathogens closely related to A. baylyi. KEYWORDS: Acinetobacter baylyi; Translation; Ribosome; Elongation Factor P; Polyproline; Biofilm; Surface Associated Motility; Antibiotic Resistance