Pathogens

Editorial

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4 pages, 213 KiB

Open AccessEditorial

Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance

by Kidon Sung, Saeed Khan and Juhee Ahn

Pathogens 2023, 12(2), 352; https://doi.org/10.3390/pathogens12020352 - 20 Feb 2023

Cited by 1 | Viewed by 2196

Abstract

Bacteria can grow either as planktonic cells or as communities within biofilms [...] Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

Research

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13 pages, 324 KiB

Open AccessArticle

Antimicrobial Resistance and Virulence Characterization of Listeria monocytogenes Strains Isolated from Food and Food Processing Environments

by Patryk Wiśniewski, Arkadiusz Józef Zakrzewski, Anna Zadernowska and Wioleta Chajęcka-Wierzchowska

Pathogens 2022, 11(10), 1099; https://doi.org/10.3390/pathogens11101099 - 25 Sep 2022

Cited by 24 | Viewed by 4010

Abstract

Listeria monocytogenes is a particularly foodborne pathogen associated with listeriosis, which can be disseminated in food and food processing environments. This study aimed to determine the serotypes and characteristics of virulence factors and antibiotic resistance among 40 L. monocytogenes strains isolated from food [...] Read more.

Listeria monocytogenes is a particularly foodborne pathogen associated with listeriosis, which can be disseminated in food and food processing environments. This study aimed to determine the serotypes and characteristics of virulence factors and antibiotic resistance among 40 L. monocytogenes strains isolated from food (n = 27) purchased in Olsztyn (Warmia and Mazury region, Poland) and food processing environments in Poland (n = 13). Isolates were assigned to serotypes 1/2a, 1/2c, 3a, and 3c using polymerase chain reaction (PCR). The results showed that serotype 1/2a (66.7%) was the most prevalent among strains from food, and serotype 1/2c (53.8%) among strains from the food processing environments. Five different virulence factors (hlyA, prfA, inlB, luxS, sigB) were detected in all isolates from the food processing environments using PCR. The hlyA (100.0%), prfA (100.0%), and inlB (96.3%) were the most prevalent in food strains. Seven (25.9%) of the strains of food and ten (76.9%) strains from the food processing environments showed the ability to form biofilm. The tested isolates were subjected to antibiotic susceptibility testing against 12 antibiotics used in the mitigation of listeriosis, using the disk diffusion method. The most frequent were intermediate resistance and resistance to clindamycin. Twelve (92.3%) strains from the food processing environments, and twenty-three (85.2%) from food were non-susceptible to clindamycin. Generally, antibacterial resistance determinants (Lde, aadB, aac(3)-IIa(aacC2)^a, penA, mefA, lnuA, lnuB, sulI, sulII) were detected in sixteen (59.0%) strains from food and four (30.8%) from the food processing environments, by PCR. The most frequent were the mefA-lnuA (n = 7; 20.0%) and lnuA (n = 6; 17.1%) genotypes. From this research, we can conclude that virulent and antimicrobial-resistant strains of L. monocytogenes are present in food and the food processing environment in Poland, which may pose a potential health risk to consumers. Monitoring for the control of virulent and antimicrobial-resistant L. monocytogenes strains in the food system can contribute to effective planning and prevention of their spread. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

18 pages, 2525 KiB

Open AccessArticle

Evidence of Virulent Multi-Drug Resistant and Biofilm-Forming Listeria Species Isolated from Various Sources in South Africa

by Christ-Donald Kaptchouang Tchatchouang, Justine Fri, Peter Kotsoana Montso, Giulia Amagliani, Giuditta Fiorella Schiavano, Madira Coutlyne Manganyi, Giulia Baldelli, Giorgio Brandi and Collins Njie Ateba

Pathogens 2022, 11(8), 843; https://doi.org/10.3390/pathogens11080843 - 27 Jul 2022

Cited by 9 | Viewed by 3647

Abstract

Listeriosis is a foodborne disease caused by Listeria monocytogenes species and is known to cause severe complications, particularly in pregnant women, young children, the elderly, and immunocompromised individuals. The aim of this study was to investigate the presence of Listeria species in food [...] Read more.

Listeriosis is a foodborne disease caused by Listeria monocytogenes species and is known to cause severe complications, particularly in pregnant women, young children, the elderly, and immunocompromised individuals. The aim of this study was to investigate the presence of Listeria species in food and water using both biochemical and species-specific PCR analysis. L. monocytogenes isolates were further screened for the presence of various antibiotic resistance, virulence, and biofilm-forming determinants profiles using phenotypic and genotypic assays. A total of 207 samples (composed of meat, milk, vegetables, and water) were collected and analyzed for presence of L. monocytogenes using species specific PCR analysis. Out of 267 presumptive isolates, 53 (19.85%) were confirmed as the Listeria species, and these comprised 26 L. monocytogenes, 3 L. innocua, 2 L. welshimeri, and 1 L. thailandensis. The remaining 21 Listeria species were classified as uncultured Listeria, based on 16SrRNA sequence analysis results. A large proportion (76% to 100%) of the L. monocytogenes were resistant to erythromycin (76%), clindamycin (100%), gentamicin (100%), tetracycline (100%), novobiocin (100%), oxacillin (100%), nalidixic acid (100%), and kanamycin (100%). The isolates revealed various multi-drug resistant (MDR) phenotypes, with E-DA-GM-T-NO-OX-NA-K being the most predominant MDR phenotypes observed in the L. monocytogenes isolates. The virulence genes prfA, hlyA, actA, and plcB were detected in 100%, 68%, 56%, and 20% of the isolates, respectively. In addition, L. monocytogenes isolates were capable of forming strong biofilm at 4 °C (%) after 24 to 72 h incubation periods, moderate for 8% isolates at 48 h and 20% at 72 h (p < 0.05). Moreover, at 25 °C and 37 °C, small proportions of the isolates displayed moderate (8–20%) biofilm formation after 48 and 72 h incubation periods. Biofilm formation genes flaA and luxS were detected in 72% and 56% of the isolates, respectively. These findings suggest that proper hygiene measures must be enforced along the food chain to ensure food safety. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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9 pages, 2708 KiB

Open AccessArticle

Antibiofilm Activity of β-Lactam/β-Lactamase Inhibitor Combination against Multidrug-Resistant Salmonella Typhimurium

by Nana Nguefang Laure and Juhee Ahn

Pathogens 2022, 11(3), 349; https://doi.org/10.3390/pathogens11030349 - 13 Mar 2022

Cited by 3 | Viewed by 2778

Abstract

This study was designed to assess the effect of β-lactam/β-lactamase inhibitor combinations on the inhibition of biofilm formation of Salmonella Typhimurium. The anti-planktonic and anti-biofilm activities of ampicillin (AMP), ceftriaxone (CEF), and combination treatments of antibiotics and sulbactam (AMP + SUL and CEF [...] Read more.

This study was designed to assess the effect of β-lactam/β-lactamase inhibitor combinations on the inhibition of biofilm formation of Salmonella Typhimurium. The anti-planktonic and anti-biofilm activities of ampicillin (AMP), ceftriaxone (CEF), and combination treatments of antibiotics and sulbactam (AMP + SUL and CEF + SUL) were evaluated against antibiotic-sensitive S. Typhimurium ATCC 19585 (ST^AS) and clinically isolated multidrug-resistant (MDR) S. Typhimurium CCARM 8009 (ST^MDR). Compared to the control, the minimum inhibitory concentrations (MICs) of AMP against ST^AS and CEF against ST^MDR were decreased from 32 to 16 μg/mL and 0.25 to 0.125 μg/mL, respectively, in the presence of SUL. The numbers of ST^MDR treated with AMP + SUL and CEF + SUL were effectively reduced by more than 2 logs after 4 h of incubation at 37 °C. The β-lactamase activities of ST^AS and ST^MDR treated with AMP and CEF were reduced from 3.3 to 2.6 μmol/min/mL and from 8.3 to 3.4 μmol/min/mL, respectively, in the presence of SUL. The biofilm cell numbers of ST^AS and ST^MDR were reduced at all treatments after 24 h of incubation at 37 °C. The biofilm cell numbers of ST^AS and ST^MDR were reduced by more than 2 logs in the presence of SUL compared to the AMP and CEF alone. The lowest relative fitness level was 0.6 in ST^AS treated with AMP + SUL, while no significant differences in the relative fitness were observed in ST^MDR. This study suggests that β-lactamase inhibitors (BLIs) could be used for controlling biofilm formation of β-lactamase-producing multidrug-resistant S. Typhimurium. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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16 pages, 3411 KiB

Open AccessArticle

Role of Efflux Pump-Mediated Antibiotic Resistance in Quorum Sensing-Regulated Biofilm Formation by Salmonella Typhimurium

by Jirapat Dawan, Yinyue Li, Feng Lu, Xinlong He and Juhee Ahn

Pathogens 2022, 11(2), 147; https://doi.org/10.3390/pathogens11020147 - 24 Jan 2022

Cited by 22 | Viewed by 4011

Abstract

This study was designed to assess the influence of efflux pump activity on the biofilm formation in Salmonella Typhimurium. Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (ST^WT) and clinically isolated S. Typhimurium CCARM 8009 (ST^CI) were treated with [...] Read more.

This study was designed to assess the influence of efflux pump activity on the biofilm formation in Salmonella Typhimurium. Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (ST^WT) and clinically isolated S. Typhimurium CCARM 8009 (ST^CI) were treated with ceftriaxone (CEF), chloramphenicol (CHL), ciprofloxacin (CIP), erythromycin (ERY), norfloxacin (NOR), and tetracycline (TET) in autoinducer-containing media in the absence and presence of phenylalanine-arginine β-naphthylamide (PAβN) to compare efflux pump activity with biofilm-forming ability. The susceptibilities of ST^WT and ST^CI were increased in the presence of PAβN. ERY+PAβN showed the highest decrease in the minimum inhibitory concentration (MIC) of ERY from 256 to 2 μg/mL against ST^WT and ST^CI. The antimicrobial activity of NOR against planktonic cells was significantly increased in the presence of PAβN, showing the lowest numbers of ST^WT (3.2 log CFU/cm²), and the TET+PAβN effectively inhibited the growth of ST^CI (5.2 log CFU/cm²). The lowest biofilm-forming abilities were observed at NOR+PAβN against ST^WT (biofilm-forming index, BFI < 0.41) and CEF+PAβN against ST^CI (BFI = 0.32). The bacteria swimming motility and relative fitness varied depending on the antibiotic and PAβN treatments. The motility diameters of ST^WT were significantly decreased by NOR+PAβN (6 mm) and TET+PAβN (15 mm), while the lowest motility of ST^CI was observed at CIP+PAβN (8 mm). The significant decrease in the relative fitness levels of ST^WT and ST^CI was observed at CIP+PAβN and NOR+PAβN. The PAβN as an efflux pump inhibitor (EPI) can improve the antimicrobial and anti-biofilm efficacy of antibiotics against S. Typhimurium. This study provides useful information for understanding the role of efflux pump activity in quorum sensing-regulated biofilm formation and also emphasizes the necessity of the discovery of novel EPIs for controlling biofilm formation by antibiotic-resistant pathogens. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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23 pages, 5681 KiB

Open AccessArticle

Antimicrobial and Anti-Biofilm Activity of Polymyxin E Alone and in Combination with Probiotic Strains of Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 against Clinical Isolates of Selected Acinetobacter spp.: A Preliminary Study

by Munaf AL-Dulaimi, Ammar Algburi, Alyaa Abdelhameed, Maria S. Mazanko, Dmitry V. Rudoy, Alexey M. Ermakov and Michael L. Chikindas

Pathogens 2021, 10(12), 1574; https://doi.org/10.3390/pathogens10121574 - 2 Dec 2021

Cited by 18 | Viewed by 3859

Abstract

Acinetobacter spp., the nosocomial pathogen, forms strong biofilms and is resistant to numerous antibiotics, causing persistent infections. This study investigates the antibacterial and anti-biofilm activity of polymyxin E alone and in combination with the cell-free supernatants (CFS) of the tested probiotic bacilli, Bacillus [...] Read more.

Acinetobacter spp., the nosocomial pathogen, forms strong biofilms and is resistant to numerous antibiotics, causing persistent infections. This study investigates the antibacterial and anti-biofilm activity of polymyxin E alone and in combination with the cell-free supernatants (CFS) of the tested probiotic bacilli, Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 against the selected Acinetobacter spp. starins. Three isolates of Acinetobacter spp., designated as Acinetobacter spp. isolate 1; Acinetobacter spp. isolate 2, and Acinetobacter spp. isolate 3, were collected from patients with burns, wounds, and blood infections, respectively. Bacterial identification and antibiotic susceptibility testing were conducted using the VITEK2 system. Auto-aggregation and coaggregation of the tested bacilli strains with the selected Acinetobacter spp. isolates were evaluated. A disk diffusion assay was used to identify the microorganism’s susceptibility to the selected antibiotics, alone and in combination with the CFS of the bacilli. The MIC and MBIC (minimum inhibitory and minimum biofilm inhibitory concentrations) of polymyxin E combined with bacilli CFS were determined. Acinetobacter spp. isolates were (i) sensitive to polymyxin E, (ii) able to form a strong biofilm, and (iii) resistant to the tested antibiotics and the CFS of tested bacilli. Significant inhibition of biofilm formation was noticed when CFS of the tested bacilli were combined with polymyxin E. The bacilli CFS showed synergy with polymyxin E against planktonic cells and biofilms of the isolated pathogens. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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Review

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18 pages, 986 KiB

Open AccessEditor’s ChoiceReview

The Use of Natural Methods to Control Foodborne Biofilms

by Michelle Marie Esposito and Sara Turku

Pathogens 2023, 12(1), 45; https://doi.org/10.3390/pathogens12010045 - 27 Dec 2022

Cited by 7 | Viewed by 3454

Abstract

Biofilms are large aggregates of various species of bacteria or other microorganisms tightly attached to surfaces through an intricate extracellular matrix. These complex microbial communities present quite the challenge in the food processing industry, as conditions such as raw meats and diverse food [...] Read more.

Biofilms are large aggregates of various species of bacteria or other microorganisms tightly attached to surfaces through an intricate extracellular matrix. These complex microbial communities present quite the challenge in the food processing industry, as conditions such as raw meats and diverse food product content in contact with workers, drains, machinery, and ventilation systems, make for prime circumstances for contamination. Adding to the challenge is the highly resistant nature of these biofilm growths and the need to keep in mind that any antimicrobials utilized in these situations risk health implications with human consumption of the products that are being processed in these locations. For that reason, the ideal means of sanitizing areas of foodborne biofilms would be natural means. Herein, we review a series of innovative natural methods of targeting foodborne biofilms, including bacteriocins, bacteriophages, fungi, phytochemicals, plant extracts, essential oils, gaseous and aqueous control, photocatalysis, enzymatic treatments, and ultrasound mechanisms. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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15 pages, 1623 KiB

Open AccessReview

Biofilm through the Looking Glass: A Microbial Food Safety Perspective

by Sapna Chitlapilly Dass and Rong Wang

Pathogens 2022, 11(3), 346; https://doi.org/10.3390/pathogens11030346 - 12 Mar 2022

Cited by 26 | Viewed by 5247

Abstract

Food-processing facilities harbor a wide diversity of microorganisms that persist and interact in multispecies biofilms, which could provide an ecological niche for pathogens to better colonize and gain tolerance against sanitization. Biofilm formation by foodborne pathogens is a serious threat to food safety [...] Read more.

Food-processing facilities harbor a wide diversity of microorganisms that persist and interact in multispecies biofilms, which could provide an ecological niche for pathogens to better colonize and gain tolerance against sanitization. Biofilm formation by foodborne pathogens is a serious threat to food safety and public health. Biofilms are formed in an environment through synergistic interactions within the microbial community through mutual adaptive response to their long-term coexistence. Mixed-species biofilms are more tolerant to sanitizers than single-species biofilms or their planktonic equivalents. Hence, there is a need to explore how multispecies biofilms help in protecting the foodborne pathogen from common sanitizers and disseminate biofilm cells from hotspots and contaminate food products. This knowledge will help in designing microbial interventions to mitigate foodborne pathogens in the processing environment. As the global need for safe, high-quality, and nutritious food increases, it is vital to study foodborne pathogen behavior and engineer new interventions that safeguard food from contamination with pathogens. This review focuses on the potential food safety issues associated with biofilms in the food-processing environment. Full article

(This article belongs to the Special Issue Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance)

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