Anti-quorum Sensing and Protective Efficacies of Naringin Against Aeromonas hydrophila Infection in Danio rerio

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.

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In vitro and in vivo efficacy of rosmarinic acid on quorum sensing mediated biofilm formation and virulence factor production in Aeromonas hydrophila

Biofouling ◽

10.1080/08927014.2016.1237220 ◽

2016 ◽

Vol 32 (10) ◽

pp. 1171-1183 ◽

Cited By ~ 24

Author(s):

Kannan Rama Devi ◽

Ramanathan Srinivasan ◽

Arunachalam Kannappan ◽

Sivasubramanian Santhakumari ◽

Murugan Bhuvaneswari ◽

...

Keyword(s):

Quorum Sensing ◽

Virulence Factor ◽

Aeromonas Hydrophila ◽

Rosmarinic Acid ◽

Biofilm Formation ◽

In Vivo Efficacy ◽

Factor Production ◽

Virulence Factor Production

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Thymol Protects Channel Catfish from Aeromonas hydrophila Infection by Inhibiting Aerolysin Expression and Biofilm Formation

Microorganisms ◽

10.3390/microorganisms8050636 ◽

2020 ◽

Vol 8 (5) ◽

pp. 636 ◽

Cited By ~ 1

Author(s):

Jing Dong ◽

Lushan Zhang ◽

Yongtao Liu ◽

Ning Xu ◽

Shun Zhou ◽

...

Keyword(s):

Quorum Sensing ◽

Channel Catfish ◽

Aeromonas Hydrophila ◽

Biofilm Formation ◽

Cell Injury ◽

Opportunistic Pathogen ◽

Protective Effects ◽

Sensing System ◽

Quorum Sensing System

Aeromonas hydrophila is an opportunistic pathogen responsible for a number of diseases in freshwater farming. Moreover, the bacterium has been identified as a zoonotic pathogen that threatens human health. Antibiotics are widely used for treatments of infectious diseases in aquaculture. However, the abuse of antibiotics has led to the emergence of antimicrobial resistant strains. Thus, novel strategies are required against resistant A. hydrophila strains. The quorum sensing (QS) system, involved in virulence factor production and biofilm formation, is a promising target in identifying novel drugs against A. hydrophila infections. In this study, we found that thymol, at sub-inhibitory concentrations, could significantly reduce the production of aerolysin and biofilm formation by inhibiting the transcription of genes aerA, ahyI, and ahyR. These results indicate that thymol inhibits the quorum sensing system. The protective effects of thymol against A. hydrophila mediated cell injury were determined by live/dead assay and lactate dehydrogenase (LDH) release assay. Moreover, the in vivo study showed that thymol could significantly decrease the mortality of channel catfish infected with A. hydrophila. Taken together, these findings demonstrate that thymol could be chosen as a phytotherapeutic candidate for inhibiting quorum sensing system-mediated aerolysin production and biofilm formation in A. hydrophila.

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N-Acylhomoserine lactones involved in quorum sensing control the type VI secretion system, biofilm formation, protease production, and in vivo virulence in a clinical isolate of Aeromonas hydrophila

Microbiology ◽

10.1099/mic.0.031575-0 ◽

2009 ◽

Vol 155 (11) ◽

pp. 3518-3531 ◽

Cited By ~ 85

Author(s):

Bijay K. Khajanchi ◽

Jian Sha ◽

Elena V. Kozlova ◽

Tatiana E. Erova ◽

Giovanni Suarez ◽

...

Keyword(s):

Quorum Sensing ◽

Aeromonas Hydrophila ◽

Biofilm Formation ◽

Secretion System ◽

Protease Production ◽

Type Vi Secretion System ◽

Double Knockout ◽

Electron Microscopic ◽

Type Vi Secretion

In this study, we delineated the role of N-acylhomoserine lactone(s) (AHLs)-mediated quorum sensing (QS) in the virulence of diarrhoeal isolate SSU of Aeromonas hydrophila by generating a double knockout ΔahyRI mutant. Protease production was substantially reduced in the ΔahyRI mutant when compared with that in the wild-type (WT) strain. Importantly, based on Western blot analysis, the ΔahyRI mutant was unable to secrete type VI secretion system (T6SS)-associated effectors, namely haemolysin coregulated protein and the valine-glycine repeat family of proteins, while significant levels of these effectors were detected in the culture supernatant of the WT A. hydrophila. In contrast, the production and translocation of the type III secretion system (T3SS) effector AexU in human colonic epithelial cells were not affected when the ahyRI genes were deleted. Solid surface-associated biofilm formation was significantly reduced in the ΔahyRI mutant when compared with that in the WT strain, as determined by a crystal violet staining assay. Scanning electron microscopic observations revealed that the ΔahyRI mutant was also defective in the formation of structured biofilm, as it was less filamentous and produced a distinct exopolysaccharide on its surface when compared with the structured biofilm produced by the WT strain. These effects of AhyRI could be complemented either by expressing the ahyRI genes in trans or by the exogeneous addition of AHLs to the ΔahyRI/ahyR+ complemented strain. In a mouse lethality experiment, 50 % attenuation was observed when we deleted the ahyRI genes from the parental strain of A. hydrophila. Together, our data suggest that AHL-mediated QS modulates the virulence of A. hydrophila SSU by regulating the T6SS, metalloprotease production and biofilm formation.

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Interaction of Acinetobacter baumannii with Human Serum Albumin: Does the Host Determines the Outcome?

Antibiotics ◽

10.3390/antibiotics10070833 ◽

2021 ◽

Vol 10 (7) ◽

pp. 833

Author(s):

Camila Pimentel ◽

Casin Le ◽

Marisel R. Tuttobene ◽

Tomas Subils ◽

Krisztina M. Papp-Wallace ◽

...

Keyword(s):

Antibiotic Resistance ◽

Human Serum Albumin ◽

Quorum Sensing ◽

Serum Albumin ◽

Human Serum ◽

Acinetobacter Baumannii ◽

Biofilm Formation ◽

Model Systems ◽

Expression Levels ◽

Multiple Drugs

Acinetobacter baumannii has become a serious threat to human health due to its extreme antibiotic resistance, environmental persistence, and capacity to survive within the host. Two A. baumannii strains, A118 and AB5075, commonly used as model systems, and three carbapenem-resistant strains, which are becoming ever more dangerous due to the multiple drugs they can resist, were exposed to 3.5% human serum albumin (HSA) and human serum (HS) to evaluate their response with respect to antimicrobial resistance, biofilm formation, and quorum sensing, all features responsible for increasing survival and persistence in the environment and human body. Expression levels of antibiotic resistance genes were modified differently when examined in different strains. The cmlA gene was upregulated or downregulated in conditions of exposure to 3.5% HSA or HS depending on the strain. Expression levels of pbp1 and pbp3 tended to be increased by the presence of HSA and HS, but the effect was not seen in all strains. A. baumannii A118 growing in the presence of HS did not experience increased expression of these genes. Aminoglycoside-modifying enzymes were also expressed at higher or lower levels in the presence of HSA or HS. Still, the response was not uniform; in some cases, expression was enhanced, and in other cases, it was tapered. While A. baumannii AB5075 became more susceptible to rifampicin in the presence of 3.5% HSA or HS, strain A118 did not show any changes. Expression of arr2, a gene involved in resistance to rifampicin present in A. baumannii AMA16, was expressed at higher levels when HS was present in the culture medium. HSA and HS reduced biofilm formation and production of N-Acyl Homoserine Lactone, a compound intimately associated with quorum sensing. In conclusion, HSA, the main component of HS, stimulates a variety of adaptative responses in infecting A. baumannii strains.

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Virulence Factors in Coagulase-Negative Staphylococci

Pathogens ◽

10.3390/pathogens10020170 ◽

2021 ◽

Vol 10 (2) ◽

pp. 170

Author(s):

Angela França ◽

Vânia Gaio ◽

Nathalie Lopes ◽

Luís D. R. Melo

Keyword(s):

Antibiotic Resistance ◽

Virulence Factors ◽

Resistance Genes ◽

Biofilm Formation ◽

Antibiotic Resistance Genes ◽

Coagulase Negative Staphylococci ◽

Healthcare Associated ◽

Major Pathogens

Coagulase-negative staphylococci (CoNS) have emerged as major pathogens in healthcare-associated facilities, being S. epidermidis, S. haemolyticus and, more recently, S. lugdunensis, the most clinically relevant species. Despite being less virulent than the well-studied pathogen S. aureus, the number of CoNS strains sequenced is constantly increasing and, with that, the number of virulence factors identified in those strains. In this regard, biofilm formation is considered the most important. Besides virulence factors, the presence of several antibiotic-resistance genes identified in CoNS is worrisome and makes treatment very challenging. In this review, we analyzed the different aspects involved in CoNS virulence and their impact on health and food.

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Attenuation of Aeromonas hydrophila Infection in Carassius auratus by YtnP, a N-acyl Homoserine Lactonase from Bacillus licheniformis T-1

Antibiotics ◽

10.3390/antibiotics10060631 ◽

2021 ◽

Vol 10 (6) ◽

pp. 631

Author(s):

Mengfan Peng ◽

Wentao Tong ◽

Zhen Zhao ◽

Ling Xiao ◽

Zhaoyue Wang ◽

...

Keyword(s):

Virulence Factors ◽

Bacillus Licheniformis ◽

Aeromonas Hydrophila ◽

Biofilm Formation ◽

Carassius Auratus ◽

Quorum Quenching ◽

Inhibition Rate ◽

Sequence Identity

In this experiment, the quorum quenching gene ytnP of Bacillus licheniformis T-1 was cloned and expressed, and the effect against infection of Aeromonas hydrophila ATCC 7966 was evaluated in vitro and vivo. The BLAST results revealed a 99% sequence identity between the ytnP gene of T-1 and its homolog in B.subtilis sub sp. BSP1, and the dendroGram showed that the similarity in the YtnP protein in T-1 was 100% in comparison with B.subtilis 3610, which was categorized as the Aidc cluster of the MBL family. The AHL lactonase activity of the purified YtnP was detected as 1.097 ± 0.7 U/mL with C6-HSL as the substrate. Otherwise, purified YtnP protein could significantly inhibit the biofilm formation of A.hydrophila ATCC 7966 with an inhibition rate of 68%. The MIC of thiamphenicol and doxycycline hydrochloride against A. hydrophila reduced from 4 μg/mL and 0.5 μg/mL to 1 μg/mL and 0.125 μg/mL, respectively, in the presence of YtnP. In addition, YtnP significantly inhibited the expression of five virulence factors hem, ahyB, ast, ep, aerA of A. hydrophila ATCC 7966 as well (p < 0.05). The results of inhibition on virulence showed a time-dependence tendency, while the strongest anti-virulence effects were within 4–24 h. In vivo, when the YtnP protein was co-injected intraperitoneally with A. hydrophila ATCC 7966, it attenuated the pathogenicity of A. hydrophila and the accumulated mortality was 27 ± 4.14% at 96 h, which was significantly lower than the average mortality of 78 ± 2.57% of the Carassius auratus injected with 108 CFU/mL of A. hydrophila ATCC 7966 only (p < 0.001). In conclusion, the AHL lactonase in B. licheniformis T-1 was proven to be YtnP protein and could be developed into an agent against infection of A. hydrophila in aquaculture.

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Influence of Glucose Concentrations on Biofilm Formation, Motility, Exoprotease Production, and Quorum Sensing in Aeromonas hydrophila

Journal of Food Protection ◽

10.4315/0362-028x.jfp-12-321 ◽

2013 ◽

Vol 76 (2) ◽

pp. 239-247 ◽

Cited By ~ 51

Author(s):

IQBAL KABIR JAHID ◽

NA-YOUNG LEE ◽

ANNA KIM ◽

SANG-DO HA

Keyword(s):

Quorum Sensing ◽

Aeromonas Hydrophila ◽

Biofilm Formation ◽

Opportunistic Pathogen ◽

Homoserine Lactone ◽

Protease Production ◽

Acyl Homoserine Lactone ◽

Human Pathogen ◽

Initial Biofilm ◽

Motility Inhibition

Aeromonas hydrophila recently has received increased attention because it is opportunistic and a primary human pathogen. A. hydrophila biofilm formation and its control are a major concern for food safety because biofilms are related to virulence. Therefore, we investigated biofilm formation, motility inhibition, quorum sensing, and exoprotease production of this opportunistic pathogen in response to various glucose concentrations from 0.05 to 2.5% (wt/vol). More than 0.05% glucose significantly impaired (P < 0.05) quorum sensing, biofilm formation, protease production, and swarming and swimming motility, whereas bacteria treated with 0.05% glucose had activity similar to that of the control (0% glucose). A stage shift biofilm assay revealed that the addition of glucose (2.5%) inhibited initial biofilm formation but not later stages. However, addition of quorum sensing molecules N-3-butanoyl-DL-homoserine lactone and N-3-hexanoyl homoserine lactone partially restored protease production, indicating that quorum sensing is controlled by glucose concentrations. Thus, glucose present in food or added as a preservative could regulate acyl-homoserine lactone quorum sensing molecules, which mediate biofilm formation and virulence in A. hydrophila.

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Inhibition of Quorum Sensing and Biofilm Formation of Esculetin on Aeromonas Hydrophila

Frontiers in Microbiology ◽

10.3389/fmicb.2021.737626 ◽

2021 ◽

Vol 12 ◽

Author(s):

Bing Sun ◽

Huaizhi Luo ◽

Huan Jiang ◽

Zhennan Wang ◽

Aiqun Jia

Keyword(s):

Quorum Sensing ◽

Aeromonas Hydrophila ◽

Biofilm Formation ◽

Laser Scanning ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Pcr Analysis ◽

Swarming Motility ◽

Gene Expression Levels ◽

Good Agreement

Quorum sensing (QS) and biofilm formation inhibition activity of esculetin on Aeromonas hydrophila SHAe 115 were evaluated. Exposure to esculetin at 25, 50, and 100μg/ml significantly inhibited the production of protease and hemolysin, the formation of biofilms and attenuated the swarming motility of A. hydrophila SHAe 115. Biofilm forming inhibition was also observed through confocal laser scanning microscopy and scanning electron microscope. Quantitative real-time PCR analysis indicated that genes positively related to QS and biofilm formation were downregulated to varying degrees, while gene (litR) negatively related to biofilm formation was significantly upregulated. The phenotypic results were in good agreement with gene expression levels. These results indicated that esculetin would be a potential QS inhibitor for A. hydrophila.

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Association among biofilm formation, virulence gene expression, and antibiotic resistance in Proteus mirabilis isolates from diarrhetic animals in northeast China

10.21203/rs.2.16502/v2 ◽

2020 ◽

Author(s):

Yadong Sun ◽

Shanshan Wen ◽

Lili Zhao ◽

Qiqi Xia ◽

Yue Pan ◽

...

Keyword(s):

Gene Expression ◽

Antibiotic Resistance ◽

Biofilm Formation ◽

Northeast China ◽

Virulence Gene ◽

Drug Susceptibility ◽

Multidrug Resistant ◽

Virulence Gene Expression ◽

Biofilm Production ◽

High Level

Abstract Background The aim of this study was to investigate the association among biofilm formation, virulence gene expression, and antibiotic resistance in P. mirabilis isolates collected from diarrhetic animals (n = 176) in northeast China between September 2014 and October 2016. Results Approximately 92.05% of the isolates were biofilm producers, whereas 7.95% of the isolates were non-producers. The prevalence of virulence genes in biofilm producers was significantly higher than that in non-producers. Biofilm production was significantly associated with the expression of ureC , zapA , rsmA , hmpA , mrpA , atfA , and pmfA ( P < 0.05). Drug susceptibility tests revealed that approximately 76.7% of the isolates were multidrug-resistant (MDR) and extensively drug-resistant (XDR). Biofilm production was significantly associated with resistance to doxycycline, tetracycline, sulfamethoxazole, kanamycin, and cephalothin ( P < 0.05). Although the pathogenicity of the biofilm producers was stronger than that of the non-producers, the biofilm-forming ability of the isolates was not significantly associated with morbidity and mortality in mice ( P > 0.05). Conclusion Our findings suggested that a high level of multidrug resistance in diarrhetic animals infected with P. mirabilis in northeast China.The results of this study indicated that the positive rates of the genes expressed by biofilm-producing P. mirabilis isolates were significantly higher than those expressed by non-producing isolates.

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Antimicrobial Resistance and Biofilm Formation of Pseudomonas aeruginosa

The International Arabic Journal of Antimicrobial Agents ◽

10.3823/846 ◽

2020 ◽

Vol 10 (2) ◽

Author(s):

Abdelraouf A Elmanama ◽

Suhaila Al-Sheboul ◽

Renad I Abu-Dan

Keyword(s):

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Quorum Sensing ◽

Bacterial Adhesion ◽

Biofilm Formation ◽

Bacterial Infections ◽

Developmental Stages ◽

Infection Site ◽

Major Virulence Factor

Abstract Pseudomonas aeruginosa threatens patient’s care. It is considered as the most complicated health care associated pathogen to be eliminated from infection site. The biofilm forming ability of P. aeruginosa, being a major virulence factor for most pathogenic microorganism, protects it from host immunity and contribute to antibiotic resistance of this organism. It is estimated that about 80% of infectious diseases are due to biofilm mode of growth. Biofilm forming ability of bacteria imparts antimicrobial resistance that leads to many persistent and chronic bacterial infections. The world is becoming increasingly under the threat of entering the “post-antibiotic era”, an era in which the rate of death from bacterial infections is higher than from cancer. This review focus on P. aeruginosa biofilm forming ability; definition, developmental stages, and significance. In addition, the quorum sensing and the antibiotic resistance of this pathogen is discussed. Keywords: Biofilm; bacterial adhesion; Pseudomonas aeruginosa; antimicrobial resistance; quorum sensing.

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