scholarly journals Prodigiosin inhibits bacterial growth and virulence factors as a potential physiological response to interspecies competition

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0253445
Author(s):  
Chee-Hoo Yip ◽  
Sobina Mahalingam ◽  
Kiew-Lian Wan ◽  
Sheila Nathan

Prodigiosin, a red linear tripyrrole pigment, has long been recognised for its antimicrobial property. However, the physiological contribution of prodigiosin to the survival of its producing hosts still remains undefined. Hence, the aim of this study was to investigate the biological role of prodigiosin from Serratia marcescens, particularly in microbial competition through its antimicrobial activity, towards the growth and secreted virulence factors of four clinical pathogenic bacteria (methicillin-resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, Salmonella enterica serovar Typhimurium and Pseudomonas aeruginosa) as well as Staphylococcus aureus and Escherichia coli. Prodigiosin was first extracted from S. marcescens and its purity confirmed by absorption spectrum, high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrophotometry (LC-MS/MS). The extracted prodigiosin was antagonistic towards all the tested bacteria. A disc-diffusion assay showed that prodigiosin is more selective towards Gram-positive bacteria and inhibited the growth of MRSA, S. aureus and E. faecalis and Gram-negative E. coli. A minimum inhibitory concentration of 10 μg/μL of prodigiosin was required to inhibit the growth of S. aureus, E. coli and E. faecalis whereas > 10 μg/μL was required to inhibit MRSA growth. We further assessed the effect of prodigiosin towards bacterial virulence factors such as haemolysin and production of protease as well as on biofilm formation. Prodigiosin did not inhibit haemolysis activity of clinically associated bacteria but was able to reduce protease activity for MRSA, E. coli and E. faecalis as well as decrease E. faecalis, Salmonella Typhimurium and E. coli biofilm formation. Results of this study show that in addition to its role in inhibiting bacterial growth, prodigiosin also inhibits the bacterial virulence factor protease production and biofilm formation, two strategies employed by bacteria in response to microbial competition. As clinical pathogens were more resistant to prodigiosin, we propose that prodigiosin is physiologically important for S. marcescens to compete against other bacteria in its natural soil and surface water environments.

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Gyan S. Sahukhal ◽  
Michelle Tucci ◽  
Hamed Benghuzzi ◽  
Gerri Wilson ◽  
Mohamed O. Elasri

Abstract Background The msaABCR operon regulates several staphylococcal phenotypes such as biofilm formation, capsule production, protease production, pigmentation, antibiotic resistance, and persister cells formation. The msaABCR operon is required for maintaining the cell wall integrity via affecting peptidoglycan cross-linking. The msaABCR operon also plays a role in oxidative stress defense mechanism, which is required to facilitate persistent and recurrent staphylococcal infections. Staphylococcus aureus is the most frequent cause of chronic implant-associated osteomyelitis (OM). The CA-MRSA USA300 strains are predominant in the United States and cause severe infections, including bone and joint infections. Results The USA300 LAC strain caused significant bone damage, as evidenced by the presence of severe bone necrosis with multiple foci of sequestra and large numbers of multinucleated osteoclasts. Intraosseous survival and biofilm formation on the K-wires by USA300 LAC strains was pronounced. However, the msaABCR deletion mutant was attenuated. We observed minimal bone necrosis, with no evidence of intramedullary abscess and/or fibrosis, along reduced intraosseous bacterial population and significantly less biofilm formation on the K-wires by the msaABCR mutant. microCT analysis of infected bone showed significant bone loss and damage in the USA300 LAC and complemented strain, whereas the msaABCR mutant’s effect was reduced. In addition, we observed increased osteoblasts response and new bone formation around the K-wires in the bone infected by the msaABCR mutant. Whole-cell proteomics analysis of msaABCR mutant cells showed significant downregulation of proteins, cell adhesion factors, and virulence factors that interact with osteoblasts and are associated with chronic OM caused by S. aureus. Conclusion This study showed that deletion of msaABCR operon in USA300 LAC strain lead to defective biofilm in K-wire implants, decreased intraosseous survival, and reduced cortical bone destruction. Thus, msaABCR plays a role in implant-associated chronic osteomyelitis by regulating extracellular proteases, cell adhesions factors and virulence factors. However additional studies are required to further define the contribution of msaABCR-regulated molecules in osteomyelitis pathogenesis.


2021 ◽  
Author(s):  
Eli COMPAORE ◽  
Moussa COMPAORE ◽  
Vincent OUEDRAOGO ◽  
Ablassé ROUAMBA ◽  
Martin KIENDREBEOGO

Abstract Background: Pseudomonas aeruginosa causes infections in human particularly immunocompromised patients with cystic fibrosis, severe burns and HIV, resulting in high morbidity and mortality. The pathogenic bacteria P aeruginosa produces virulence factors regulated by the mechanism called quorum sensing system. Objective: The aim of this study was to assess the anti-quorum sensing activity of Ageratum conyzoides extracts Method: Chromobacterium violaceum reporter strain CV026 was used to highlight any interference with bacterium QS and strains derived from P. aeruginosa PAO1 were used to reveal any interference with the expression of quorum sensing genes, and to assess any impact of extract on the kinetics of the production of pyocyanin, elastases and biofilm formation. Results: Hydro-methanolic extract at the sub-inhibitory concentration of 100 μg/mL reduced quorum sensing virulence factors production such as, pyocyanin, elastases, rhamnolipids and biofilm formation in P. aeruginosa PAO1 after 18 hours monitoring. Extract showed significant inhibition in HSL-mediated violacein production on C. violaceum CV026 after 48 hours monitoring. Biofilm formation was inhibited up to 32%. It affected QS gene expression in PAO1. The regulatory genes lasR / rhlR and the lasI synthases were most affected. At 8hours, hydro-methanolic extract reduced both QS gene to more than 30% (lasI/lasR and rhlI/R respectively 33.8% /30.2% and 36% /33.2%). RhlA and lasB genes have been relatively affected (13.4% and 28.9%). After 18 h, this extract reduced significantly the expression of regulatory 30 genes lasR (31%) and rhlR (39.6%) although synthases genes seemed to be less affected (lasI/21.2% and rhlI/11.6%). A limited impact was observed on the downstream genes (lasB /20.0% and rhlA /15.3%). No negative impact was observed on CV026 and PAO1 growth and cell viability. Our study also showed that A. conyzoides having ample amount of phenolics, flavonoids and triterpenoids. This phytochemical content could be one of the factors for showing anti quorum potential. Conclusion: Results indicate that hydro methanol 80 % extract from A. conyzoides could be a source of potential QS inhibition compounds.


2018 ◽  
Author(s):  
◽  
Johmari Logtenberg

Background: Healthcare-associated infections (HCAIs) are a global concern as they affect millions of people worldwide. Poor hygiene practices and the use of microbial contaminated medical equipment by healthcare workers (HCWs) are common contributing factors to the development of HCAIs, which result in additional hospital costs, prolonged hospital stays, development of antibacterial resistance and increases in mortality and morbidity. Because Chiropractic students (CSs) at the Durban University of Technology Chiropractic Day Clinic (DUT CDC) make use of diagnostic equipment during their consultations with patients, this study aimed to determine if the stethoscopes and sphygmomanometers that were used in the DUT CDC served as reservoirs for bacterial growth, including antibiotic-resistant bacteria, and to correlate the findings with the hygiene practices of CSs and the efficacy of disinfectants. Method: This quantitative study comprised of two phases: a phase one pre-test post-test design and a phase two cross-sectional descriptive questionnaire design. Phase one required the collection of bacterial samples from the stethoscopes and sphygmomanometers of 29 CSs before and after performing the physical assessments on new patients (58 samples). These bacterial samples were incubated and analysed. The bacterial isolates were enumerated, identified and, where appropriate, tested for antibiotic-resistance. The modified AOAC use dilution method was used to test the efficacy of the selected disinfectants. Phase two required 29 CSs to complete the research questionnaire. The data were initially captured onto Excel spreadsheets and subsequently analysed using IBM SPSS version 24.0 (p-value <0.05 was considered statistically significant), with the application of Spearman’s rank correlation, an one-way ANOVA evaluation, Tukey post hoc and paired t-tests. Results: Although the majority of the CSs was knowledgeable and regarded disinfection as important, only 13.8% applied adequate disinfection practices. The most common reasons that were stated were inadequate education or training, forgetfulness, lack of time, and disinfectant unavailability. Bacterial growth was present on 96.6% and 100.0% of the pre-test and post-test stethoscope samples respectively, and on 94.8% and 100.0% of the pre- and post-test sphygmomanometer samples respectively. The total colony-forming units (CFUs) for both the post- test readings were higher compared to their respective pre-test samples. Paired t-tests indicated significantly (p< 0.01) higher mean values for the post-stethoscope group only, with a greater distribution of the total CFUs for stethoscope samples at the diaphragms’ edge. The bacteria that were isolated from both sets of pre- and post-test samples consisted predominantly of coagulase negative staphylococci (CoNS), Micrococcus spp. and Staphylococcus aureus, while the minority consisted of Bacillus spp., Corynebacterium spp., coliforms and Escherichia coli. Overall, the majority of the bacteria was considered potentially pathogenic, except for the post-test sphygmomanometer sample. The disinfectant efficacy testing revealed results that were in contrast with the literature, which caused the researcher to question the validity and reliability of the modified AOAC use dilution method in this study. Chloramphenicol was the most effective antibiotic with a bacterial susceptibility rate of 95.7%, (Ciprofloxacin (93.2%), Vancomycin (80.8%), Amoxicillin (AMO) (69.5%) and Erythromycin (57.7%)). The resistance of Micrococcus spp. isolates to the various antibiotics was of concern. The high resistance levels of CoNS and S. aureus to AMO suggests the presence of methicillin-resistant Staphylococcus aureus and methicillin-resistant CoNS isolates. The most susceptible specie in general was Micrococcus spp. at 60.9%, followed by S. aureus at 59.6%, E. coli at 53.8%, and CoNS at 48.7%, while the least susceptible was coliforms at 36.9%. No correlations (p> 0.05) were identified between the mean CFU isolates from the CSs’ equipment and their reported average disinfection rate. Conclusions and recommendations: Both stethoscopes and sphygmomanometers were contaminated with non-pathogenic and potential pathogenic bacteria (some were resistant to multiple antibiotic classes). Although knowledgeable about equipment disinfection procedures, only 13.8% of the CSs reported disinfecting their pieces of equipment after examining every patient. The provision of adequate equipment disinfection education, the placement of visual reminders and accessible disinfectants will assist in improving the practice of adequate equipment disinfection. Moreover, equipment disinfection before and after every patient consultation will minimise cross- contamination and thus the risk of the development of HCAIs.


PHARMACON ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 834
Author(s):  
Eunike Pelealu ◽  
Defny S. Wewengkang ◽  
Surya Sumantri Abdullah

ABSTRACTSponges are one of the biota components that make up coral reefs which are quite widely distributed. The metabolite content in the sponge can ward off and inhibit the pathogenic bacteria that interfere with it. This study aims to determine the activity of inhibiting bacterial growth from the extract and fraction of Leucetta chagosensis sponge against the growth of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. The samples were extracted using the maceration method with 95% ethanol solvent and then fractionated using 3 solvents with different polarity levels, namely methanol, n-hexane and chloroform. Activity test using the disk diffusion agar method of Kirby and Bauer. Only the MeOH fraction was able to inhibit the growth of E. coli bacteria with an average inhibition zone of 6.88 mm. Whereas for S.aureus bacteria extracts and all fractions showed activity to inhibit bacterial growth with an average inhibition zone of EtOH (6.61 mm), CHCI3 (6.68 mm), n-hexane (7.83 mm) and MeOH (8.00 mm), respectively. All activities that are shown are categorized as weak (weak).Keywords: Antibacterial, Leucetta chagosensis, Staphylococcus aureus, Escherichia coli ABSTRAKSpons merupakan salah satu komponen biota penyusun terumbu karang yang penyebarannya cukup luas. Kandungan metabolit yang ada di dalam spons dapat menangkal dan menghambat bakteri patogen pengganggunya.  Penelitian ini bertujuan untuk melihat aktivitas menghambat pertumbuhan bakteri dari ekstrak dan fraksi spons Leucetta chagosensis. terhadap pertumbuhan bakteri Gram positif Staphylococcus aureus dan Gram negatif Escherichia coli. Sampel di ekstraksi menggunakan metode maserasi dengan pelarut etanol 95% lalu di fraksinasi dengan menggunakan 3 pelarut dengan tingkat kepolaran yang berbeda yaitu metanol, n-heksan dan kloroform. Uji aktivitas menggunakan metode disk diffusion agar Kirby dan Bauer.  Hanya fraksi MeOH yang mampu menghambat pertumbuhan bakteri E.coli dengan zona hambat rata-rata 6,88 mm. Sedangkan terhadap bakteri S.aureus ekstrak dan semua fraksi menunjukan aktifitas menghambat pertumbuhan bakteri dengan rata-rata zona hambat masing-masing EtOH (6,61 mm), CHCl3 (6,68 mm), n-Heksan (7,83 mm), dan MeOH (8,00 mm). Semua aktivitas yang ditunjukan dikategorikan lemah (weak).Kata kunci : Antibakteri, Leucetta chagosensis, Staphylococcus aureus,  Escherichia coli


2019 ◽  
Author(s):  
Brittney D. Gimza ◽  
Maria I. Larias ◽  
Bridget G. Budny ◽  
Lindsey N. Shaw

AbstractA primary function of the extracellular proteases of Staphylococcus aureus is to control the progression of infection by selectively modulating the stability of virulence factors. Consequently, a regulatory network exists to titrate protease abundance/activity, to influence accumulation, or lack thereof, of individual virulence factors. Herein, we comprehensively map this system, exploring regulation of the four protease loci by known and novel factors. In so doing, we determine that seven major elements (SarS, SarR, Rot, MgrA, CodY, SaeR, and SarA) form the primary network of control, with the latter three being the most powerful. We note that expression of aureolysin is largely repressed by these factors, whilst the spl operon is subject to the strongest upregulation of any protease loci, particularly by SarR and SaeR. Furthermore, when exploring scpA expression, we find it to be profoundly influenced in opposing fashions by SarA (repressor) and SarR (activator). We also present the screening of >100 regulator mutants of S. aureus, identifying 7 additional factors (ArgR2, AtlR, MntR, Rex, XdrA, Rbf, and SarU) that form a secondary circuit of protease control. Primarily these elements serve as activators, although we reveal XdrA as a new repressor of protease expression. With the exception or ArgR2, each of the new effectors appear to work through the primary network of regulation to influence protease production. Collectively, we present a comprehensive regulatory circuit that emphasizes the complexity of protease regulation and suggest that its existence speaks to the importance of these enzymes to S. aureus physiology and pathogenic potential.ImportanceThe complex regulatory role of the proteases necessitates very tight coordination and control of their expression. Whilst this process has been well studied, a major oversight has been the consideration of proteases as a single entity, rather than 10 enzymes produced from four different promoters. As such, in this study we comprehensively characterized the regulation of each protease promoter, discovering vast differences in the way each protease operon is controlled. Additionally, we broaden the picture of protease regulation using a global screen to identify novel loci controlling protease activity, uncovering a cadre of new effectors of protease expression. The impact of these elements on the activity of proteases and known regulators was characterized producing a comprehensive regulatory circuit that emphasizes the complexity of protease regulation in Staphylococcus aureus.


Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 980
Author(s):  
Tyler J. Gerken ◽  
Marilyn C. Roberts ◽  
Philip Dykema ◽  
Geoff Melly ◽  
Darren Lucas ◽  
...  

Staphylococcus aureus are human facultative pathogenic bacteria and can be found as contaminants in the environment. The aim of our study was to determine whether methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) isolated from coastal beach and river waters, anchialine pools, sand, and wastewater on the island of Hawaiʻi, Hawaiʻi, are a potential health risk. Samples were collected from three regions on Hawaiʻi Island from July to December 2020 during the COVID-19 pandemic and were characterized using whole-genome sequencing (WGS). From WGS data, multilocus sequence typing (MLST), SCCmec type, antimicrobial resistance genes, virulence factors, and plasmids were identified. Of the 361 samples, 98.1% were positive for Staphylococcus spp. and 7.2% were S. aureus positive (n = 26); nine MRSA and 27 MSSA strains were characterized; multiple isolates were chosen from the same sample in two sand and seven coastal beach water samples. The nine MRSA isolates were multi-drug resistant (6–9 genes) sequence type (ST) 8, clonal complex (CC) 8, SCCmec type IVa (USA300 clone), and were clonally related (0–16 SNP differences), and carried 16–19 virulence factors. The 27 MSSA isolates were grouped into eight CCs and 12 STs. Seventy-eight percent of the MSSA isolates carried 1–5 different antibiotic resistance genes and carried 5–19 virulence factors. We found S. aureus in coastal beach and river waters, anchialine pools, and sand at locations with limited human activity on the island of Hawaiʻi. This may be a public health hazard.


2005 ◽  
Vol 52 (3) ◽  
pp. 633-638 ◽  
Author(s):  
Adam Dubin ◽  
Paweł Mak ◽  
Grzegorz Dubin ◽  
Małgorzata Rzychon ◽  
Justyna Stec-Niemczyk ◽  
...  

The increasing antibiotic resistance of pathogenic bacteria calls for the development of alternative antimicrobial strategies. Possible approaches include the development of novel, broad-spectrum antibiotics as well as specific targeting of individual bacterial virulence factors. It is impossible to decide currently which strategy will prove more successful in the future since they both promise different advantages, but also introduce diverse problems. Considering both approaches, our laboratory's research focuses on the evaluation of hemocidins, broad-spectrum antibacterial peptides derived from hemoglobin and myoglobin, and staphostatins, specific inhibitors of staphopains -- Staphylococcus aureus secreted proteases that are virulence factors regarded as possible targets for therapy. The article summarizes recent advances in both fields of study and presents perspectives for further development and possible applications.


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