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Staphylococcus aureus is widely recognized as a highly hazardous pathogen that poses significant threats to food safety and public health. This study aimed to assess the prevalence, antimicrobial resistance, and genetic characteristics of S. aureus isolates recovered from 288 frozen flour and rice product samples in Shanghai, China, between September 2019 and May 2020. A total of 81 S. aureus isolates were obtained, representing 25 sequence types (STs), with ST7 being the most prevalent (17.28%, n = 14). The majority of S. aureus isolates (85.19%, n = 69) carried at least one enterotoxin gene, with the seg gene being the most frequently detected (51.85%, n = 42). Additionally, 12 isolates (14.81%) were identified as methicillin-resistant S. aureus (MRSA) through mecA gene detection. Notably, this study reported the presence of an ST398 MRSA isolate in frozen flour and rice products for the first time. All MRSA isolates displayed multidrug resistance, with the highest resistance observed against cefoxitin (100.00%), followed by penicillin (91.67%) and erythromycin (66.67%). Genomic analysis of the 12 MRSA isolates revealed the presence of twenty distinct acquired antimicrobial resistance genes (ARGs), eight chromosomal point mutations, and twenty-four unique virulence genes. Comparative genome analysis indicated close genetic relationships between these MRSA isolates and previously reported MRSA isolates from clinical infections, highlighting the potential transmission of MRSA through the food chain and its implications for public health. Significantly, the identification of three plasmids harboring ARGs, insertion sequences (ISs), the origin of transfer site (oriT), and the relaxase gene suggested the potential for horizontal transfer of ARGs via conjugative plasmids in S. aureus. In conclusion, this study revealed significant contamination of retail frozen flour and rice products with S. aureus, and provided essential data for ensuring food safety and protecting public health.

Biofilms enable pathogenic bacteria to survive in unfavorable environments. As biofilm-forming pathogens can cause rapid food spoilage and recurrent infections in humans, especially their presence in the food industry is problematic. Using chemical disinfectants in the food industry to prevent biofilm formation raises serious health concerns. Further, the ability of biofilm-forming bacterial pathogens to tolerate disinfection procedures questions the traditional treatment methods. Thus, there is a dire need for alternative treatment options targeting bacterial pathogens, especially biofilms. As clean-label products without carcinogenic and hazardous potential, natural compounds with growth and biofilm-inhibiting and biofilm-eradicating potentials have gained popularity as natural preservatives in the food industry. However, the use of these natural preservatives in the food industry is restricted by their poor availability, stability during food processing and storage. Also there is a lack of standardization, and unattractive organoleptic qualities. Nanotechnology is one way to get around these limitations and as well as the use of underutilized bioactives. The use of nanotechnology has several advantages including traversing the biofilm matrix, targeted drug delivery, controlled release, and enhanced bioavailability, bioactivity, and stability. The nanoparticles used in fabricating or encapsulating natural products are considered as an appealing antibiofilm strategy since the nanoparticles enhance the activity of the natural products against biofilms of foodborne bacterial pathogens. Hence, this literature review is intended to provide a comprehensive analysis of the current methods in nanotechnology used for natural products delivery (biofabrication, encapsulation, and nanoemulsion) and also discuss the different promising strategies employed in the recent and past to enhance the inhibition and eradication of foodborne bacterial biofilms.

Staphylococcus aureus is a versatile pathogen known to cause hospital- and community-acquired, foodborne, and zoonotic infections. The clinical infections by S. aureus cause an increase in morbidity and mortality rates and treatment costs, aggravated by the emergence of drug-resistant strains. As a multi-faceted pathogen, it is imperative to consolidate the knowledge on its pathogenesis, including the mechanisms of virulence regulation, development of antimicrobial resistance, and biofilm formation, to make it amenable to different treatment strategies. Nanomaterials provide a suitable platform to address this challenge, with the potential to control intracellular parasitism and multidrug resistance where conventional therapies show limited efficacy. In a nutshell, the first part of this review focuses on the impact of S. aureus on human health and the role of virulence factors and biofilms during pathogenesis. The second part discusses the large diversity of nanoparticles and their applications in controlling S. aureus infections, including combination with antibiotics and phytochemicals and the incorporation of antimicrobial coatings for biomaterials. Finally, the limitations and prospects using nanomaterials are highlighted, aiming to foster the development of novel nanotechnology-driven therapies against multidrug-resistant S. aureus.

Biofilm-related nosocomial infections may cause a wide range of life-threatening infections. In this regard, Pseudomonas aeruginosa biofilm is becoming a serious health burden due to its capability to develop resistance to natural and synthetic drugs. The utilization of nanoparticles that inhibit biofilm formation is one of the major strategies to control infections caused by biofilm-forming pathogens. Hydroxyapatite (HA) is a synthetic ceramic material having properties similar to natural bones. Herein, a co-precipitation method followed by microwave treatment was used to synthesize HA nanoparticles (HANPs). The resulting HANPs were characterized using X-ray diffraction and transmission electron microscopy. Then, their antibiofilm properties against P. aeruginosa ATCC 10145 were examined in vitro. The needle-shaped HANPs were 30 and 90 nm long in width and length, respectively. The synthesized HANPs inhibited the biofilm formation of P. aeruginosa ATCC 10145 in a concentration-dependent manner, which was validated by light and confocal laser scanning microscopy. Hence, this study demonstrated that HANPs could be used to control the biofilm-related infections of P. aeruginosa.

Natural alternatives replacing artificial additives have gained much attention in the consumer’s view because of the growing search for clean label products that are devoid of carcinogenic and toxic effects. Plant polyphenols are considered as suitable alternative natural preservatives with antioxidant and antimicrobial properties. However, their uses in the food industry are undermined by a series of limitations such as low solubility and stability during food processing and storage, lack of standardization, and undesirable organoleptic properties. Different approaches in the use of polyphenols have been proposed in order to overcome the current hurdles related to food preservation. This review article specifically focuses on the antibacterial activity of plant-derived polyphenols as well as their applications as food preservatives, main challenges, and other trends in the food industry.

Aim Staphylococcus aureus causes several complicated infections. Despite decades of research on developing new antimicrobials, methicillin-resistant S. aureus (MRSA) remains a global health problem. Hence, there is a dire need to identify potent natural antibacterial compounds as an alternative to antimicrobials. In this light, the present work divulges the antibacterial efficacy and the action mechanism of 2-hydroxy-4-methoxybenzaldehyde (HMB) isolated from Hemidesmus indicus against S. aureus. Methods and Results Antimicrobial activity of HMB was assessed. HMB exhibited 1024 µg ml−1 as the minimum inhibitory concentration (MIC) and 2 × MIC as the minimum bactericidal concentration against S. aureus. The results were validated by spot assay, time kill, and growth curve analysis. In addition, HMB treatment increased the release of intracellular proteins and nucleic acid contents from MRSA. Additional experiments assessing the structural morphology of bacterial cells using SEM analysis, β-galactosidase enzyme activity, and the fluorescence intensities of propidium iodide and rhodamine123 dye divulged that the cell membrane as one of the targets of HMB to hinder S. aureus growth. Moreover, the mature biofilm eradication assay revealed that HMB dislodged nearly 80% of the preformed biofilms of MRSA at the tested concentrations. Further, HMB treatment was found to sensitize MRSA cells upon combining tetracycline treatment. Conclusions The present study suggests that HMB is a promising compound with antibacterial and antibiofilm activities and could act as a lead structure for developing new antibacterial drugs against MRSA. [ABSTRACT FROM AUTHOR]

Quorum sensing (QS) is a signaling mechanism governed by bacteria used to converse at inter- and intra-species levels through small self-produced chemicals called N-acylhomoserine lactones (AHLs). Through QS, bacteria regulate and organize the virulence factors' production, including biofilm formation. AHLs can be degraded by an action called quorum quenching (QQ) and hence QQ strategy can effectively be employed to combat biofilm-associated bacterial pathogenesis. The present study aimed to identify novel bacterial species with QQ potential. Screening of Palk Bay marine sediment bacteria for QQ activity ended up with the identification of marine bacterial isolate 28 (MSB-28), which exhibited a profound QQ activity against QS biomarker strain Chromobacterium violaceum ATCC 12472. The isolate MSB-28 was identified as Psychrobacter sp. through 16S-rRNA sequencing. Psychrobacter sp. also demonstrated a pronounced activity in controlling the biofilm formation in different bacteria and biofilm-associated virulence factors' production in P. aeruginosa PAO1. Solvent extraction, heat inactivation, and proteinase K treatment assays clearly evidence the enzymatic nature of the bioactive lead. Furthermore, AHL's lactone ring cleavage was confirmed with experiments including ring closure assay and chromatographic analysis, and thus the AHL-lactonase enzyme production in Psychrobacter sp. To conclude, this is the first report stating the AHL-lactonase mediated QQ activity from marine sediment bacteria Psychrobacter sp. Future work deals with the characterization, purification, and mass cultivation of the purified protein and should pave the way to assessing the feasibility of the identified protein in controlling QS and biofilm-mediated multidrug resistant bacterial infections in mono or multi-species conditions. [ABSTRACT FROM AUTHOR]

It is now well known that the quorum sensing (QS) mechanism coordinates the production of several virulence factors and biofilm formation in most pathogenic microorganisms. Aeromonas hydrophila is a prime pathogen responsible for frequent outbreaks in aquaculture settings. Recent studies have also continuously reported that A. hydrophila regulates virulence factor production and biofilm formation through the QS system. In addition to the presence of antibiotic resistance genes, biofilm-mediated antibiotic resistance increases the severity of A. hydrophila infections. To control the bacterial pathogenesis and subsequent infections, targeting the QS mechanism has become one of the best alternative methods. Though very few compounds were identified as QS inhibitors against A. hydrophila , to date, the screening and identification of new and effective natural QS inhibitors is a dire necessity to control the infectious A. hydrophila. The present study endorses naringin (NA) as an anti-QS and anti-infective agent against A. hydrophila. Initially, the NA showed a concentration-dependent biofilm reduction against A. hydrophila. Furthermore, the results of microscopic analyses and quantitative virulence assays displayed the promise of NA as a potential anti-QS agent. Subsequently, the downregulation of ahh1 , aerA , lip and ahyB validate the interference of NA in virulence gene expression. Furthermore, the in vivo assays were carried out in zebrafish model system to evaluate the anti-infective potential of NA. The outcome of the immersion challenge assay showed that the recovery rate of the zebrafish has substantially increased upon treatment with NA. Furthermore, the quantification of the bacterial load upon NA treatment showed a decreased level of bacterial counts in zebrafish when compared to the untreated control. Moreover, the NA treatment averts the pathogen-induced histoarchitecture damages in vital organs of zebrafish, compared to their respective controls. The current study has thus analyzed the anti-QS and anti-infective capabilities of NA and could be employed to formulate effective treatment measures against A. hydrophila infections. [ABSTRACT FROM AUTHOR]

Candida albicans is considered an exclusive etiologic agent of candidiasis, a very common fungal infection in human. The expression of virulence factors contributes highly to the pathogenicity of C. albicans. These factors include biofilm formation, yeast-to-hyphal transition, adhesins, aspartyl proteases, and phospholipases secretion. Moreover, resistance development is a critical issue for the therapeutic failure of antifungal agents against systemic candidiasis. To circumvent resistance development, the present study investigated the virulence targeted therapeutic activity of the phyto-bioactive compound morin against C. albicans. Morin is a natural compound commonly found in medicinal plants and widely used in the pharmaceutical and cosmetic products/industries. The present study explicated the significant inhibitory potential of morin against biofilm formation and other virulence factors' production, such as yeast-hyphal formation, phospholipase, and exopolymeric substances, in C. albicans. Further, qPCR analysis confirmed the downregulation of biofilm and virlence-related genes in C. albicans upon morin treatment, which is in correspondence with the in vitro bioassays. Further, the docking analysis revealed that morin shows strong affinity with Hwp-1 protein, which regulates the expression of biofilm and hyphal formation in C. albicans and, thereby, abolishes fungal pathogenicity. Moreover, the anti-infective potential of morin against C. albicans -associated systemic candidiasis is confirmed through an in vivo approach using biomedical model organism zebrafish (Danio rerio). The outcomes of the in vivo study demonstrate that the morin treatment effectively rescues animals from C. albicans infections and extends their survival rate by inhibiting the internal colonization of C. albicans. Histopathology analysis revealed extensive candidiasis-related pathognomonic changes in the gills, intestine, and kidney of animals infected with C. albicans , while no extensive abnormalities were observed in morin-treated animals. The results evidenced that morin has the ability to protect against the pathognomonic effect and histopathological lesions caused by C. albicans infection in zebrafish. Thus, the present study suggests that the utilization of morin could act as a potent therapeutic medication for C. albicans instigated candidiasis. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis (SE) is an opportunistic nosocomial pathogen that accounts for recalcitrant device-related infections worldwide. Owing to the growing interest in plants and their secondary metabolites targeting bacterial adhesion, this study was intended to uncover the anti-biofilm potential of Hemidesmus indicus and its major constituent 2-hydroxy-4-methoxybenzaldehyde (HMB) against SE. The minimum biofilm inhibitory concentration (MBIC) of H. indicus root extract and HMB were found to be 500 and 250 µg ml−1, respectively. The results of time-dependent biofilm inhibition and mature biofilm disruption assays confirmed that HMB targets initial cell adhesion. Furthermore, interference by HMB in the expression of adhesin genes (icaA, aap and bhp) and biofilm components was associated with an increased susceptibility of SE to oxidative stress and antibiotics. To conclude, this study reports for the first time HMB as a potential drug against SE biofilms. [ABSTRACT FROM AUTHOR]

Abstract In humans, the occurrence of bacterial communities in the form of biofilm is considered as a major intrinsic factor accountable for a variety of stubborn infections. Staphylococcus aureus and S. epidermidis have gained considerable attention in clinical settings owing to the formation of intractable and long-lasting biofilms in medical device. The current study has been designed to explain the biofilm inhibitory efficacy of geraniol and cefotaxime combination (GCC) against S. epidermidis and methicillin-resistant S. aureus (MRSA). Biofilm biomass quantification assay was performed to evaluate the antibiofilm activity of GCC against S. epidermidis and MRSA. The minimal biofilm inhibitory concentration of GCC was found to be 100 μg/ml of geraniol and 2 μg/ml of cefotaxime. Further, microscopic analyses ascertained the devastating potential of GCC on the test pathogens' biofilm formation. Besides biofilm inhibition, GCC also suppressed the production of extracellular polymeric substance, slime and staphyloxanthin. More, GCC significantly increased the susceptibility of the test pathogens towards human blood. Further, the results of real time PCR analysis and in vivo assay using Caenorhabditis elegans unveiled the anti-biofilm potentials of GCC. Thus, the present study demonstrates the significant use of polytherapy treatment approaches to overcome the biofilm associated infections of Staphylococcus spp. [ABSTRACT FROM AUTHOR]

Urinary tract infections are the utmost common bacterial infections caused by Proteus mirabilis, Pseudomonas aeruginosa, Escherichia coli, and Serratia marcescens. These uropathogens resist the action of several antibiotics due to their ability to form biofilms. Most of these bacterial pathogens use the quorum sensing (QS) machinery to co-ordinate their cells and regulate several virulence factors and biofilm formation. On the other hand, the anti-quorum sensing (anti-QS) and antibiofilm potential of silver nanoparticles have been well reported against certain bacterial pathogens, but to the best of our knowledge, no report is available against the pathogenicity of uropathogens in particular S. marcescens and P. mirabilis. Therefore, the present study is primarily focused on the anti-QS and antibiofilm potential of Piper betle-based synthesized silver nanoparticles (PbAgNPs) against S. marcescens and P. mirabilis. Initially, the silver nanoparticles were synthesized by the aqueous extract of P. betle and characterized by UV-absorbance spectroscopy, XRD, FT-IR, SEM, TEM, and DLS. The synthesized silver nanoparticles were assessed for their anti-QS activity and the obtained results revealed that the PbAgNPs inhibited the QS-mediated virulence factors such as prodigiosin, protease, biofilm formation, exopolysaccharides and hydrophobicity productions in uropathogens. The gene expression analysis divulged the downregulation of fimA, fimC, flhD, and bsmB genes in S. marcescens and flhB, flhD, and rsbA genes in P. mirabilis, respectively. The in vivo Caenorhabditis elegans assays revealed the non-toxic and anti-adherence efficiency of PbAgNPs. Furthermore, the non-toxic effect of PbAgNPs was also confirmed through peripheral blood mononuclear cells and normal lung epithelial cells. Therefore, the contemporary study demonstrates the use of PbAgNPs as a possible alternative toward conventional antibiotics in controlling QS and biofilm-related uropathogen infections. [ABSTRACT FROM AUTHOR]

Rosmarinic acid (RA) was assessed for its quorum sensing inhibitory (QSI) potential against Aeromonas hydrophila strains AH 1, AH 12 and MTCC 1739. The pathogenic strains of A. hydrophila were isolated from infected zebrafish and identified through biochemical analysis and amplification of a species- specific gene (rpsL). The biofilm inhibitory concentration (BIC) of RA against A. hydrophila strains was found to be 750 µg ml-1. At this concentration, RA reduced the QS mediated hemolysin, lipase and elastase production in A. hydrophila. In FT-IR analysis, RA treated A. hydrophila cells showed a reduction in cellular components. Gene expression analysis confirmed the down-regulation of virulence genes such as ahh1, aerA, lip and ahyB. A. hydrophila infected zebrafish upon treatment with RA showed increased survival rates. Thus, the present study demonstrates the use of RA as a plausible phytotherapeutic compound to control QS mediated biofilm formation and virulence factor production in A. hydrophila. [ABSTRACT FROM AUTHOR]

The prevalence of antimicrobial resistance reduces the effectiveness of antimicrobial drugs in preventing and treating infectious diseases caused by pathogenic organisms, such as bacteria, fungi, and viruses. Because of the burgeoning growth of microbes with antimicrobial-resistant traits, there is a dire need to identify and develop novel and effective antimicrobial agents to treat infections from antimicrobial-resistant strains. The marine environment is rich in ecological biodiversity and can be regarded as an untapped resource for prospecting novel bioactive compounds. Therefore, exploring the marine environment for antimicrobial agents plays a significant role in drug development and biomedical research. Several earlier scientific investigations have proven that bacterial diversity in the marine environment represents an emerging source of structurally unique and novel antimicrobial agents. There are several reports on marine bacterial secondary metabolites, and many are pharmacologically significant and have enormous promise for developing effective antimicrobial drugs to combat microbial infections in drug-resistant pathogens. In this review, we attempt to summarize published articles from the last twenty-five years (1996–2020) on antimicrobial secondary metabolites from marine bacteria evolved in marine environments, such as marine sediment, water, fauna, and flora. [ABSTRACT FROM AUTHOR]

Aggregated mode of bacterial growth in the form of biofilm is a prime reason for nonspecific antibiotic resistance and several persistent and chronic infections in clinical settings. Hence, targeting the initial adherence, early & mature biofilms of the bacterial pathogens would be a suitable alternative to control multidrug-resistant bacterial infections. The root bark of Hemidesmus indicus is a well-known in the Ayurvedic medicinal system. In the present study, antibiofilm potential of methanolic extract of H. indicus root (HI) was evaluated against nosocomial bacterial pathogens such as methicillin-resistant and -susceptible Staphylococcus aureus and Streptococcus pyogenes. The minimum biofilm inhibitory concentration (MBIC) of HI was found to be 300 and 150 μg/ml for S. aureus and S. pyogenes , respectively. Further results from microscopic analyses and in vitro bioassays validated the biofilm inhibitory potential of HI. HI at MBIC failed to eradicate the preformed biofilms of the test pathogens. Chromatographic analysis divulges the presence of several functional metabolites in the HI responsible for its bioactivity. Thus, the present study suggests that HI targets the biofilm formation of the test pathogens by hindering the initial adherence of the cells to form biofilm. The extract of HI root can be used as a suitable alternate to treat biofilm mediated infection and would be considered in drug formulations to control the onset of biofilm mediated bacterial infections. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Piper betle , a tropical creeper plant belongs to the family Piperaceae . The leaves of this plant have been well known for their therapeutic, religious and ceremonial value in South and Southeast Asia. It has also been reported to possess several biological activities including antimicrobial, antioxidant, antinociceptive, antidiabetic, insecticidal and gastroprotective activities and used as a common ingredient in indigenous medicines. In Indian system of ayurvedic medicine, P. betle has been well recognized for its antiseptic properties and is commonly applied on wounds and lesions for its healing effects. Aim of the study To evaluate the anti-quorum sensing (anti-QS) and antibiofilm efficacy of P. betle and its bioactive metabolite phytol against Serratia marcescens . Materials and methods The P. betle ethyl acetate extract (PBE) was evaluated for its anti-QS efficacy against S. marcescens by assessing the prodigiosin and lipase production at 400 and 500 µg ml −1 concentrations. In addition, the biofilm biomass quantification assay was performed to evaluate the antibiofilm activity of PBE against S. marcescens . Besides, the influence of PBE on bacterial biofilm formation was assessed through microscopic techniques. The biofilm related phenomenons like exopolysaccharides (EPS) production, hydrophobicity and swarming motility were also examined to support the antibiofilm activity of PBE. Transcriptional analysis of QS regulated genes in S. marcescens was also done. Characterization of PBE was done by separation through column chromatography and identification of active metabolites by gas chromatography -mass spectrometry. The major compounds of active fractions such as hexadecanoic acid, eugenol and phytol were assessed for their anti-QS activity against S. marcescens . Further, the in vitro bioassays such as protease, biofilm and HI quantification were also carried out to confirm the anti-QS and antibiofilm potential of phytol in PBE. Results PBE inhibits QS mediated prodigiosin pigment production in S. marcescens, which confirmed its anti-QS potential against S. marcescens . At 500 µg ml −1 concentration, PBE significantly inhibited the production of protease, lipase, biofilm and EPS to the level of 71%, 68%, 65% and 43% in S. marcescens , respectively. Further, their antibiofilm efficacy was confirmed through microscopic techniques. In addition, PBE effectively inhibited the hydrophobicity and swarming motility. Additionally, the results of qPCR analysis validated the downregulation of QS genes. Chromatographic techniques the presence of hexadecanoic acid, eugenol and phytol in PBE and the potential bioactive compound with anti-QS activity was identified as phytol. In vitro assays with phytol evidenced the potent inhibition of QS-controlled prodigiosin, protease, biofilm and hydrophobicity in S. marcescens , without exerting any deleterious effect on its growth. Conclusion This study demonstrates the promising anti-QS and antibiofilm activities of PBE and its active metabolite phytol, and confirms the ethnopharmacological applications of these leaves against S. marcescens infections. [ABSTRACT FROM AUTHOR]

• Rice water was selected as the cheapest carbon source for formulating rice water medium in order to optimize biosurfactant synthesis by Pseudomonas aeruginosa. • The in vitro and in vivo antifungal efficacy and biocontrol strategy of the biosurfactant against Fusarium wilt of Abelmoschus esculentus were demonstrated. • The identification of the rhl gene, TLC, FT-IR, HPLC, and GC–MS analyses suggested the biosurfactant was rhamnolipid. • The biosurfactant that has been produced will aid in large-scale commercialization and will replace the usage of chemical surfactants in agricultural wastes. Biosurfactants, in comparison to chemical surfactants, have a greater impact on various applications due to their stability, degradability, and other physicochemical characteristics. Increasing yield and lowering production costs are critical to enhancing biosurfactant efficiency. Because biosurfactant production costs are determined by the least expensive substrate, agro-based industrial waste is one of the most promising economic strategies. Therefore, in this study we aimed to achieve the biosurfactant production in Pseudomonas aeruginosa by low cost rice medium using Response surface methodology (RSM) and assessed their competence in controlling the Fusarium wilt of Abelmoschus esculentus. Initially, the selected strain P. aeruginosa PBS29 was confirmed using 16S rRNA sequencing. Then, the 20% (v/v) of rice water was chosen as the cheapest carbon source for formulating rice water medium to optimize biosurfactant production using Central Composite Design. Enhanced biosurfactant yield of 9.35 g/l was attained through RSM by 0.59 fold higher than preliminary analysis. The model was significant with a regression coefficient of 0.98. The optimal condition was identified as 1.18% (w/v) nitrogen source [glutamic acid], pH 6.8, temperature at 37.4 °C, 2.5% (v/v) inoculum size, and 167.9 rpm agitation. In-vitro antifungal activity and biocontrol strategy of the biosurfactant demonstrated against Fusarium wilt of Abelmoschus esculentus at 100 μg/ml concentration by both soil drenching and foliar spray by pot trial. Furthermore, the detection of rhl gene, TLC, FT-IR, HPLC and GC–MS analyses suggested that the biosurfactant is rhamnolipid. Overall, in this study, the biosurfactant was synthesized using inexpensive rice water and substantiated its safe use as a biocontrol agent, which will help large-scale commercialization and interchange chemical surfactant used in agricultural wastes. [ABSTRACT FROM AUTHOR]

Quorum Sensing (QS) mechanism, a bacterial density-dependent gene expression system, governs the Serratia marcescens pathogenesis through the production of virulence factors and biofilm formation. The present study demonstrates the anti-quorum sensing (anti-QS), antibiofilm potential and in vivo protective effect of phytol, a diterpene alcohol broadly utilized as food additive and in therapeutics fields. In vitro treatment of phytol (5 and 10 μg/ml) showed decreasing level of biofilm formation, lipase and hemolysin production in S. marcescens compared to their respective controls. More, microscopic analyses confirmed the antibiofilm potential of phytol. The biofilm related phenomenons such as swarming motility and exopolysccharide productions were also inhibited by phytol. Furthermore, the real-time analysis elucidated the molecular mechanism of phytol which showed downregulation of fimA, fimC, flhC, flhD, bsmB, pigP , and shlA gene expressions. On the other hand, the in vivo rescue effect of phytol was assessed against S. marcescens associated acute pyelonephritis in Wistar rat. Compared to the infected and vehicle controls, the phytol treated groups (100 and 200 mg/kg) showed decreased level of bacterial counts in kidney, bladder tissues and urine samples on the 5th post infection day. As well, the phytol treatment showed reduced level of virulence enzymes such as lipase and protease productions compared to the infected and vehicle controls. Further, the infected and vehicle controls showed increasing level of inflammatory markers such as malondialdehyde (MDA), nitric oxide (NO) and myeloperoxidase (MPO) productions. In contrast, the phytol treatment showed decreasing level of inflammatory markers. In histopathology, the uninfected animal showed normal kidney and bladder structure, wherein, the infected animals showed extensive infiltration of neutrophils in kidney and bladder tissues. In contrast, the phytol treatment showed normal kidney and bladder tissues. Additionally, the toxic effect of phytol (200 mg/kg) was assessed by single dose toxicity analysis. No changes were observed in hematological, biochemical profiles and histopathological analysis of vital organs in phytol treated animals compared to the untreated controls. Hence, this study suggested the potential use of phytol for its anti-QS, antibiofilm and anti-inflammatory properties against S. marcescens infections and their associated inflammation reactions.

Rosmarinic acid (RA) was assessed for its quorum sensing inhibitory (QSI) potential against Aeromonas hydrophila strains AH 1, AH 12 and MTCC 1739. The pathogenic strains of A. hydrophila were isolated from infected zebrafish and identified through biochemical analysis and amplification of a species-specific gene (rpsL). The biofilm inhibitory concentration (BIC) of RA against A. hydrophila strains was found to be 750 μg ml -1 . At this concentration, RA reduced the QS mediated hemolysin, lipase and elastase production in A. hydrophila. In FT-IR analysis, RA treated A. hydrophila cells showed a reduction in cellular components. Gene expression analysis confirmed the down-regulation of virulence genes such as ahh1, aerA, lip and ahyB. A. hydrophila infected zebrafish upon treatment with RA showed increased survival rates. Thus, the present study demonstrates the use of RA as a plausible phytotherapeutic compound to control QS mediated biofilm formation and virulence factor production in A. hydrophila.