Category Archives: Biofilm

Research – Nanoparticle Coatings on Glass Surfaces to Prevent Pseudomonas fluorescens AR 11 Biofilm Formation

MDPI

Abstract

Microbial colonization of surfaces is a sanitary and industrial issue for many applications, leading to product contamination and human infections. When microorganisms closely interact with a surface, they start to produce an exo-polysaccaridic matrix to adhere to and protect themselves from adverse environmental conditions. This type of structure is called a biofilm. The aim of our work is to investigate novel technologies able to prevent biofilm formation by surface coatings. We coated glass surfaces with melanin-ZnO2, melanin-TiO2, and TiO2 hybrid nanoparticles. The functionalization was performed using cold plasma to activate glass-substrate-coated surfaces, that were characterized by performing water and soybean oil wetting tests. A quantitative characterization of the antibiofilm properties was done using Pseudomonas fluorescens AR 11 as a model organism. Biofilm morphologies were observed using confocal laser scanning microscopy and image analysis techniques were used to obtain quantitative morphological parameters. The results highlight the efficacy of the proposed surface coating to prevent biofilm formation. Melanin-TiO2 proved to be the most efficient among the particles investigated. Our results can be a valuable support for future implementation of the technique proposed here in an extended range of applications that may include further testing on other strains and other support materials.

Research – The Role of Biofilms in the Pathogenesis of Animal Bacterial Infections

MDPI

Abstract

Biofilms are bacterial aggregates embedded in a self-produced, protective matrix. The biofilm lifestyle offers resilience to external threats such as the immune system, antimicrobials, and other treatments. It is therefore not surprising that biofilms have been observed to be present in a number of bacterial infections. This review describes biofilm-associated bacterial infections in most body systems of husbandry animals, including fish, as well as in sport and companion animals. The biofilms have been observed in the auditory, cardiovascular, central nervous, digestive, integumentary, reproductive, respiratory, urinary, and visual system. A number of potential roles that biofilms can play in disease pathogenesis are also described. Biofilms can induce or regulate local inflammation. For some bacterial species, biofilms appear to facilitate intracellular invasion. Biofilms can also obstruct the healing process by acting as a physical barrier. The long-term protection of bacteria in biofilms can contribute to chronic subclinical infections, Furthermore, a biofilm already present may be used by other pathogens to avoid elimination by the immune system. This review shows the importance of acknowledging the role of biofilms in animal bacterial infections, as this influences both diagnostic procedures and treatment.

Research – Bacterial biofilm reduction by 275 and 455 nm light pulses emitted from light emitting diodes

Wiley Online

Biofilm of antibiotic resistant bacteria. Rod-shaped and spherical bacteria. Escherichia coli, Pseudomonas, Mycobacterium tuberculosis, Klebsiella, Staphylococcus aureus, MRSA. 3D illustration

Abstract

Eradication of biofilms from the food contact surfaces is a challenging task, owing to their increased resistance to sanitizers and regular cleaning practices. The treatment with the light pulses emitted from the light emitting diode (LED) is an emerging surface decontamination technology, that can produce the antibiofilm effect by photodynamic inactivation. The objective of this study was to understand the antibiofilm efficacy of the 275 (Ultraviolet-C [UV-C]) and 455 nm (Blue) light pulses emitted from the LEDs against single and dual-species biofilms of Salmonella Typhimurium ATCC13311 and Aeromonas australiensis 03-09 on stainless steel (SS) coupons formed at different time. The biofilm formation by S. Typhimurium was improved when grown with A. australiensis in dual-species culture. Both 275 and 455 nm light pulses showed significant antibiofilm activity against S. Typhimurium and A. australiensis in single and dual-species biofilms. For instance, the 275 nm LED treatment of surfaces of SS coupons with 1.8 J/cm2 dose on each surface, produced reductions of 4.24 and 3.9 log (CFU/cm2) in single (cell attachment) and dual-species biofilms of S. Typhimurium, and reductions of 4.45 and 4.99 log (CFU/cm2) in single and dual-species biofilms of A. australiensis. However, the susceptibility of A. australiensis toward 455 nm LED treatments was influenced by the presence of S. Typhimurium in the dual-species biofilm. The confocal laser scanning microscopy images revealed significant cell membrane damage in the dual-species biofilms by the LED treatments with 275 and 455 nm light pulses. Overall, several factors like surface temperature increase, strains used, treatment dose, treatment time, and incubation period of biofilms influenced the inactivation efficacy of the 275 and 455 nm LED treatments against the biofilms formed on SS coupons. This study provides an insight into the inactivation efficacy of LED light pulses against bacterial biofilms on food grade SS surfaces.

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

MDPI

Bacteria can grow either as planktonic cells or as communities within biofilms. The biofilm growth mode is the dominant lifestyle of most bacterial species and 40–80% of microorganisms are associated with biofilms [1]. Biofilm is a sessile community that is irreversibly attached to a substratum or interface or to other members of the community [2]. It is surrounded by extracellular polymeric substances (EPS) that include extracellular polysaccharides, extracellular DNA, lipids, proteins, and other elements [3]. Biofilm formation is a complex but well-regulated process that can be classified into five distinct stages [4]. In the first stage, planktonic bacteria attach to a surface. Salmonella species, Listeria monocytogenesCampylobacter jejuni, or Escherichia coli have specific structures on the surface of the bacteria, such as flagella, curli, fimbriae, and pili, which help the bacteria attach [5].
The second stage is the adhesion step, which includes an initial reversible adhesion resulting in loose adhesion and a subsequent irreversible adhesion resulting in more stable adhesion. The third stage is to secrete EPS and form microcolonies. This is followed by biofilm maturation, which produces large amounts of EPS to grow in size and build three-dimensional structures. The final stage is the stage in which the biofilm is dispersed, releasing the planktonic cells and initiating the formation of a new biofilm at another location.
Microbial cells living within biofilms are protected from various environmental stresses such as desiccation, osmotic changes, oxidative stress, metal toxicity, radiation, antibiotics, disinfectants, and the host immune system [6]. Biofilms are much less sensitive to antimicrobial agents than planktonic cells, and several mechanisms contribute to their resistance to antimicrobials [7]. The exopolysaccharide matrix prevents the entry of antimicrobial agents by reducing diffusion and acting as a primary barrier [8]. Most antimicrobial agents kill rapidly dividing cells more effectively, but slow growth of biofilms leads to resistance [9]. Changes in metabolic activity within biofilms, genetic changes of antimicrobial resistant determinants in target cells, extrusion of antimicrobial agents using efflux pumps, and the presence of persistent cells also contribute to antimicrobial resistance [10].

Research – Exploring Possible Ways to Enhance the Potential and Use of Natural Products through Nanotechnology in the Battle against Biofilms of Foodborne Bacterial Pathogens

MDPI

Abstract

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.

Research – Microbial Dynamics in Mixed-Culture Biofilms of Salmonella Typhimurium and Escherichia coli O157:H7 and Bacteria Surviving Sanitation of Conveyor Belts of Meat Processing Plants

MDPI

Abstract

Biofilm formation can lead to the persistence of Salmonella Typhimurium (ST) and E. coli O157:H7 (O157). This study investigated the impact of meat processing surface bacteria (MPB) on biofilm formation by O157 (non-biofilm former; NF) and ST (strong biofilm former; BF). MPB were recovered from the contacting surfaces (CS), non-contacting surfaces (NCS), and roller surfaces (RS) of a beef plant conveyor belt after sanitation. O157 and ST were co-inoculated with MPB (CO), or after a delay of 48 h (IS), into biofilm reactors containing stainless steel coupons and incubated at 15 °C for up to 144 h. Coupons were withdrawn at various intervals and analyzed by conventional plating and 16S rRNA gene amplicon sequencing. The total bacterial counts in biofilms reached approximately 6.5 log CFU/cm2, regardless of MPB type or development mode. The mean counts for O157 and ST under equivalent conditions mostly did not differ (p > 0.05), except for the IS set at 50 h, where no O157 was recovered. O157 and ST were 1.6 ± 2.1% and 4.7 ± 5.0% (CO) and 1.1 ± 2.2% and 2.0 ± 2.8% (IS) of the final population. Pseudomonas dominated the MPB inocula and biofilms, regardless of MPB type or development mode. Whether or not a pathogen is deemed BF or NF in monoculture, its successful integration into complex multi-species biofilms ultimately depends on the presence of certain other residents within the biofilm.

Research – Antibacterial and Anti-Biofilm Properties of Diopside Powder Loaded with Lysostaphin

MDPI

Abstract

Background: Diopside-based ceramic is a perspective biocompatible material with numerous potential applications in the field of bone prosthetics. Implantable devices and materials are often prone to colonization and biofilm formation by pathogens such as Staphylococcus aureus, which in the case of bone grafting leads to osteomyelitis, an infectious bone and bone marrow injury. To lower the risk of bacterial colonization, implanted materials can be impregnated with antimicrobials. In this work, we loaded the antibacterial enzyme lysostaphin on diopside powder and studied the antibacterial and antibiofilm properties of such material to probe the utility of this approach for diopside-based prosthetic materials. Methods: Diopside powder was synthesized by the solid-state method, lysostaphin was loaded on diopside by adsorption, the release of lysostaphin from diopside was monitored by ELISA, and antibacterial and anti-biofilm activity was assessed by standard microbiological procedures. Results and conclusions: Lysostaphin released from diopside powder showed high antibacterial activity against planktonic bacteria and effectively destroyed 24-h staphylococcal biofilms. Diopside-based materials possess a potential for the development of antibacterial bone grafting materials.

Research – The Use of Natural Methods to Control Foodborne Biofilms

MDPI

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 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.

Research – Exploring the Diversity of Biofilm Formation by the Food Spoiler Brochothrix thermosphacta

MDPI

Abstract

Brochothrix thermosphacta is considered as a major spoiler of meat and seafood products. This study explores the biofilm formation ability and the biofilm structural diversity of 30 multi-origin B. thermosphacta strains using a set of complementary biofilm assays (biofilm ring test, crystal violet staining, and confocal laser scanning microscopy). Two major groups corresponding to low and high biofilm producers were identified. High biofilm producers presented flat architectures characterized by high surface coverage, high cell biovolume, and high surface area.

Research – Life and death of an ‘altruistic’ bacterium

Science Daily

Biofilms, complex communities of bacteria, abound around us: on the surface of cheese where they give off flavors and aromas, in streams where they form the slimy substance on rocks, on our teeth where they form plaque.

Living in a biofilm provides numerous advantages to bacteria: things like resource sharing, shelter from predators, and increased resistance to toxic compounds such as antibiotics.

But having the option to leave the biofilm when environmental conditions deteriorate can be a plus for bacteria, too, allowing them to relocate to a more hospitable environment.

”For the bacterium Caulobacter crescentus, the biofilm becomes a kind of prison in perpetuity: once cells are attached to a surface through a strong adhesive at one end of the cell, they cannot leave the biofilm,” said Yves Brun, a professor in the Department of Microbiology, Infectious Diseases and Immunology at Université de Montréal.

”However, when these attached cells divide, their unattached ‘daughter’ cells have a choice of joining the biofilm or swimming away.”