Antibacterial potential of essential oils against planktonic and sessile cells of Escherichia coli isolated from diarrhea cases in swine

Escherichia coli is a pathogen associated with infections in piglets in the post-weaning phase, its pathogenicity is related to the animal's susceptibility to bacterial enterotoxins. The objective of the present study was to determine the EOs activity against E. coli strain, in the form planktonic and sessile. Although the Disc-Diffusion tests to determine the Minimum Inhibitory Concentration, do not fully corroborate with the other analyzes of this study, it was noticed bacteria inhibition. The EOs were prepared at 0.4%, 0.8% and 1.0% for tests. The tested EOs were effective against E. coli planktonic cells (p<0.05). As for the sessile cells, the most significant result was inhibition and 100% sessile cells at the concentration of 1.0% of Cymbopogon citratus EO. Although there was resistance in some treatments, the tested EOs demonstrated inhibition capacity, constituting promising alternatives for the control of E. coli, especially of planktonic cells.

Optimization of Propidium Monoazide qPCR (Viability-qPCR) to Quantify the Killing by the Gardnerella-Specific Endolysin PM-477, Directly in Vaginal Samples from Women with Bacterial Vaginosis

Quantification of the number of living cells in biofilm or after eradication treatments of biofilm, is problematic for different reasons. We assessed the performance of pre-treatment of DNA, planktonic cells and ex vivo vaginal biofilms of Gardnerella with propidium monoazide (PMAxx) to prevent qPCR-based amplification of DNA from killed cells (viability-qPCR). Standard PMAxx treatment did not completely inactivate free DNA and did not affect living cells. While culture indicated that killing of planktonic cells by heat or by endolysin was complete, viability-qPCR assessed only log reductions of 1.73 and 0.32, respectively. Therefore, we improved the standard protocol by comparing different (combinations of) parameters, such as concentration of PMAxx, and repetition, duration and incubation conditions of treatment. The optimized PMAxx treatment condition for further experiments consisted of three cycles, each of: 15 min incubation on ice with 50 µM PMAxx, followed by 15 min-long light exposure. This protocol was validated for use in vaginal samples from women with bacterial vaginosis. Up to log2.2 reduction of Gardnerella cells after treatment with PM-477 was documented, despite the complex composition of the samples, which might have hampered the activity of PM-477 as well as the quantification of low loads by viability-qPCR.

Bactericidal efficacy of slightly acidic electrolyzed water (SAEW) against Listeria monocytogenes planktonic cells and biofilm on food-contact surfaces

In the present study, the bactericidal efficacy of slightly acidic electrolyzed water (SAEW) against L. monocytogenes planktonic cells and biofilm on food-contact surfaces including stainless steel and glass was systematically evaluated. The results showed that SAEW (pH of 5.09 and available chlorine concentration (ACC) of 60.33 mg/L) could kill L. monocytogenes on food-contact surfaces completely in 30 s, whose disinfection efficacy is equal to that of NaClO solutions (pH of 9.23 and ACC of 253.53 mg/L). The results showed that long exposure time and high ACC contributed to the enhancement of the disinfection efficacy of SAEW on L. monocytogenes on food-contact surfaces. Moreover, the log reduction of SAEW treatment presented an increasing tendency within the prolonging of treatment time when SAEW was used to remove the L. monocytogenes biofilm formed on stainless steel and glass surfaces, which suggested that SAEW could remove L. monocytogenes biofilm effectively and its disinfection efficacy is equal to (in case of stainless steel) or higher than (in case of glass) that of high ACC of NaClO solutions. In addition, the results of the crystal violet staining and scanning electron microscopy (SEM) also demonstrated that SAEW treatment could remove the L. monocytogenes biofilm on food-contact surfaces.

The Effect of Antibiotics on Planktonic Cells and Biofilm Formation Ability of Collected Arcobacter-like Strains and Strains Isolated within the Czech Republic

The purpose of this study was to test the in vitro effects of ampicillin, ciprofloxacin, clindamycin, erythromycin, gentamicin, and tetracycline on planktonic cells of Arcobacter-like microorganisms and on their biofilm formation ability. The minimum inhibitory concentrations (MICs) were determined by the microdilution method. Further, biofilm formation ability in the presence of various concentrations of antibiotics was evaluated by a modified Christensen method. Most of the 60 strains exhibited high susceptibility to gentamicin (98.3%), ciprofloxacin (95.0%), and erythromycin (100.0%). High level of resistance was observed to clindamycin and tetracycline with MIC50 and MIC90 in range of 4–32 mg/L and 32–128 mg/L, respectively. Combined resistance to both clindamycin and tetracycline was found in 38.3% of tested strains. In general, higher biofilm formation was observed especially at lower concentrations of antibiotics (0.13–2 mg/L). However, a significant decrease in biofilm formation ability of Pseudarcobacter defluvii LMG 25694 was exhibited with ampicillin and clindamycin at concentrations above 32 or 8 mg/L, respectively. Biofilm formation represents a potential danger of infection and also a risk to human health, in particular due to antimicrobial-resistant strains and the ability to form a biofilm structure at a concentration that is approximately the MIC determined for planktonic cells.

Photodynamic Effect of 5,10,15,20-Tetrakis[4-(3-N,N-dimethylaminopropoxy)phenyl]chlorin towards the Human Pathogen Candida albicans under Different Culture Conditions

Photocytotoxic activity sensitized by 5,10,15,20-tetrakis[4-(3-N,N-dimethylaminopropoxy)phenyl]chlorin (TAPC) was investigated in Candida albicans under different culture conditions. Planktonic cells incubated with 2.5 μM TAPC were eradicated after 5 min irradiation with white light. Studies in the presence of reactive oxygen species scavengers indicated the involvement of mainly a type II mechanism. Furthermore, cell growth of C. albicans was suppressed in the presence of 5 μM TAPC. A decrease in pseudohyphae survival of 5 log was found after 30 min irradiation. However, the photokilling of this virulence factor reached a 1.5 log reduction in human serum. The uptake of TAPC by pseudohyphae decreased in serum due to the interaction of TAPC with albumin. The binding constant of the TAPC-albumin complex was ~104 M−1, while the bimolecular quenching rate constant was ~1012 s−1 M−1, indicating that this process occurred through a static process. Thus, the photoinactivation of C. albicans was considerably decreased in the presence of albumin. A reduction of 2 log in cell survival was observed using 4.5% albumin and 30 min irradiation. The results allow optimizing the best conditions to inactivate C. albicans under different culture conditions.

Antimicrobial Resistance in Pseudomonas aeruginosa Biofilms

Pseudomonas aeruginosa (PA) is part of a group of common nosocomial pathogens that exhibit multidrug resistance, thus proving to be a significant threat to healthcare. This study analyzes the ability of four commonly used antibiotics to observe eradication of the PA biofilm growth. Ceftazidime (CAZ), Tobramycin (TOB), Ofloxacin (OFLX), Meropenem (MEM), were tested against overnight cultures of PA strain PA01. The minimal inhibitory concentrations (MIC) of planktonic cells for all the four antibiotics were determined using broth microdilution while the minimal bactericidal concentrations (MBCs) were determined by colony count after antibiotic treatment and regrowth. Biofilm growth inhibition was performed by treating cells with antibiotic at the time of inoculation while eradication was determined by adding antibiotics 24 hours after inoculation, allowing mature biofilm formation, followed by the measurement of absorbance. PA planktonic cells exhibited the highest susceptibility to MEM compared to overnight grown PA biofilm which demonstrated resistance to CAZ, complete sensitivity to ofloxacin, and minimal sensitivity to TOB and MEM. PA biofilm displayed dose-dependent sensitivity to TOB, MEM and OFLX, and a significant level of resistance to CAZ during the inhibition phase. However, in the eradication phase, PA showed significant resistance to TOB followed by CAZ while PA biofilm showed sensitivity at higher concentrations of MEM. Our study exhibits that PA strain PA01 is resistant to ceftazidime in both planktonic and biofilm phases. While ofloxacin proved to be the most effective even at lower concentrations when compared with other antibiotics, tobramycin was most effective at higher concentrations for eradicating and inhibiting PA biofilms.

Biofilm formation and antagonistic activity of Lacticaseibacillus rhamnosus (PTCC1712) and Lactiplantibacillus plantarum (PTCC1745)

Currently, the health benefits of probiotic bacteria are well known, and this has taken up a great deal of space in food science and health, both research and operational. On the other hand, anti-biofilm properties on food pathogens in the food and pharmaceutical industries have created an attractive challenge. This study aimed to describe the inhibitory activity of cell-free supernatants (CFS), planktonic cells, and biofilm form of lactobacilus strains (L. rhamnosus and L. plantarum) against food pathogens such as Pseudomonas aeruginosa and Listeria monocytogenes. Anti-bacterial activities of the CFS of lactobacillus strains were assessed by the microplate method and via violet staining. Evaluation of the antagonistic activity of planktonic cells and biofilm of LAB were performed by the spread plate method. The results showed the incubation time of 48 h was the best time to produce biofilm. Although the planktonic states reduce the pathogens bacterial about 1 –1.5 log, but in biofilm forms, decreased L. monocytogenes about 4.5 log compared to the control, and in the case of P. aeruginosa, a growth reduction of about 2.13 log was observed. Furthermore, biofilm formation of L. monocytogenes in the presence of L. rhamnosus cell-free supernatant was more weakly than L. plantarum CFS, but their CFS effect on reducing the bacterial population of P. aeruginosa was the same. According to the study, biofilm produced by probiotic strains can be considered a new approach for biological control. Also, cell-free supernatant can be used as postbiotic in the food and pharmaceutical industries.