Contribution of horizontal gene transfer to the functionality of microbial biofilm on a macroalgae

Horizontal gene transfer (HGT) is thought to be an important driving force for microbial evolution and niche adaptation and has been show in vitro to occur frequently in biofilm communities. However, the extent to which HGT takes place and what functions are being transferred in more complex and natural biofilm systems remains largely unknown. To address this issue, we investigated here HGT and enrichment of gene functions in the biofilm community of the common kelp (macroalgae) Ecklonia radiata in comparison to microbial communities in the surrounding seawater. We found that HGTs in the macroalgal biofilms were dominated by transfers between bacterial members of the same class or order and frequently involved genes for nutrient transport, sugar and phlorotannin degradation as well as stress responses, all functions that would be considered beneficial for bacteria living in this particular niche. HGT did not appear to be driven by mobile gene elements, indicating rather an involvement of unspecific DNA uptake (e.g. natural transformation). There was also a low overlap between the gene functions subject to HGT and those enriched in the biofilm community in comparison to planktonic community members. This indicates that much of the functionality required for bacteria to live in an E. radiata biofilm might be derived from vertical or environmental transmissions of symbionts. This study enhances our understanding of the relative role of evolutionary and ecological processes in driving community assembly and genomic diversity of biofilm communities.Subject terms: Biofilms, Metagenomics     

Impact of genomics on microbial food safety

Genome sequences are now available for many of the microbes that cause food-borne diseases. The information contained in pathogen genome sequences, together with the development of themed and whole-genome DNA microarrays and improved proteomics techniques, might provide tools for the rapid detection and identification of such organisms, for assessing their biological diversity and for understanding their ability to respond to stress. The genomic information also provides insight into the metabolic capacity and versatility of microbes; for example, specific metabolic pathways might contribute to the growth and survival of pathogens in a range of niches, such as food-processing environments and the human host. New concepts are emerging about how pathogens function, both within foods and in interactions with the host. The future should bring the first practical benefits of genome sequencing to the field of microbial food safety, including strategies and tools for the identification and control of emerging pathogens.     

Removal and transfer of viruses on food contact surfaces by cleaning cloths

Contamination of food contact surfaces with pathogens is considered an important vehicle for the indirect transmission of food-borne diseases. Five different cleaning cloths were assessed for the ability to remove viruses from food contact surfaces (stainless steel surface and nonporous solid surface) and to transfer viruses back to these surfaces. Cleaning cloths evaluated include two different cellulose/cotton cloths, one microfiber cloth, one nonwoven cloth, and one cotton terry bar towel. Four viral surrogates (murine norovirus [MNV], feline calicivirus [FCV], bacteriophages PRD1 and MS2) were included. Removal of FCV from stainless steel was significantly greater (P ≤ 0.05) than that from nonporous solid surface, and overall removal of MNV from both surfaces was significantly less (P ≤ 0.05) than that of FCV and PRD1. Additionally, the terry towel removed significantly fewer total viruses (P ≤ 0.05) than the microfiber and one of the cotton/cellulose cloths. The cleaning cloth experiments were repeated with human norovirus. For transfer of viruses from cloth to surface, both cellulose/cotton cloths and microfiber transferred an average of 3.4 and 8.5 total PFU, respectively, to both surfaces, and the amounts transferred were significantly different (P ≤ 0.05) from those for the nonwoven cloth and terry towel (309 and 331 total PFU, respectively). There was no statistically significant difference (P > 0.05) in the amount of virus transfer between surfaces. These data indicate that while the cleaning cloths assessed here can remove viruses from surfaces, some cloths may also transfer a significant amount of viruses back to food contact surfaces.     

Impact of a synbiotic food on the gut microbial ecology and metabolic profiles

Background  

The human gut harbors a diverse community of microorganisms which serve numerous important functions for the host wellbeing. Functional foods are commonly used to modulate the composition of the gut microbiota contributing to the maintenance of the host health or prevention of disease. In the present study, we characterized the impact of one month intake of a synbiotic food, containing fructooligosaccharides and the probiotic strains Lactobacillus helveticus Bar13 and Bifidobacterium longum Bar33, on the gut microbiota composition and metabolic profiles of 20 healthy subjects.     

Effect of surface roughness, biofilm coverage and biofilm structure on the electrochemical efficiency of microbial cathodes

Biofilms of Geobacter sulfurreducens were formed under chronoamperometry at −0.5 V and −0.6 V vs. Ag/AgCl on stainless steel cathodes and tested for fumarate reduction. Increasing the surface roughness Ra from 2.0 μm to 4.0 μm increased currents by a factor of 1.6. The overall current density increased with biofilm coverage. When the current density was calculated with respect to the biofilm-coated area only, values up to 280 A/m² were derived. These values decreased with biofilm coverage and indicated that isolated cells or small colonies locally provide higher current density than dense colonies. Steel composition affected the current values because of differences in biofilm structure and electron transfer rates. Biofilms formed under polarisation revealed better electrochemical characteristics than biofilm developed at open circuit. This work opens up new guidelines for the design of microbial cathodes: a uniform carpet of isolated bacteria or small colonies should be targeted, avoiding the formation of large colonies.     

Impact of prescribed cleaning and disinfectant use on microbial contamination in the home

Aims: To identify and quantify the presence of Escherichia coli, Staphylococcus aureus, Salmonella, hepatitis A and norovirus in households and to assess the effect of chlorine and quaternary ammonium–based disinfectants following a prescribed use. Methods and Results: Eleven sites distributed in kitchen, bathroom, pet and children′s areas of two groups of 30 homes each: (i) a nonprescribed disinfectant user group and (ii) a disinfectant protocol user group. During the 6‐week study, samples were collected once a week except for week one when sample collection occurred immediately before and after disinfectant application to evaluate the disinfectant protocol. The concentration and occurrence of bacteria were less in the households with prescribed use of disinfectants. The greatest reductions were for E. coli (99%) and Staph. aureus (99·9999%), respectively. Only two samples were positive for HAV, while norovirus was absent. Disinfection protocols resulted in a significant (P < 0·05) microbial reduction in all areas of the homes tested compared to homes not using a prescribed protocol. Conclusions: The study suggests that disinfectant product application under specific protocol is necessary to achieve greater microbial reductions. Significance and Impact of the Study: Prescribed protocols constitute an important tool to reduce the occurrence of potential disease‐causing micro‐organisms in households.     

Availability of glucose and light modulates the structure and function of a microbial biofilm

We have studied the differences in the organic matter processing and biofilm composition and structure between autoheterotrophic and heterotrophic biofilm communities. Microbial communities grown on artificial biofilms were monitored, following incubation under light and dark conditions and with or without the addition of glucose as a labile organic compound. Glucose addition greatly affected the microbial biofilm composition as shown by differences in 16S rRNA gene fingerprints. A significant increase in β-glucosidase and peptidase enzyme activities were also observed in glucose-amended biofilms incubated in the dark, suggesting an active bacterial community. Light enhanced the algal and bacterial growth, as well as higher extracellular enzyme activity, thereby indicating a tight algal–bacterial coupling in biofilms incubated under illumination. In these biofilms, organic compounds excreted by photosynthetic microorganisms were readily available for bacterial heterotrophs. This algal–bacterial relationship weakened in glucose-amended biofilms grown in the light, probably because heterotrophic bacteria preferentially use external labile compounds. These results suggest that the availability of labile organic matter in the flowing water and the presence of light may alter the biofilm composition and function, therefore affecting the processing capacity of organic matter in the stream ecosystem.     

A biofilm model developed to investigate survival and disinfection of Mycobacterium mucogenicum in potable water

Water in healthcare environments can be a source for healthcare-associated infections (HAI). However, information on the exposure risk to opportunistic pathogens in potable water distribution systems (PWDS) is lacking. Laboratory studies characterizing the interaction of opportunistic pathogens with biofilms are needed to understand their role in water systems within healthcare facilities. A stable, repeatable, PWDS multi-species biofilm model comprising Sphingomonas paucimobilis, Methylobacterium sp., Delftia acidovorans, and Mycobacterium mucogenicum was developed in the CDC Biofilm Reactor (CBR), reaching 6 log₁₀ CFU cm^?2 within 6 days. The model was used to investigate the interaction of the opportunistic pathogen M. mucogenicum with the other species, and to determine the efficacy of monochloramine (NH₂Cl) as a disinfectant against 2-week-old biofilms. Addition of 1 or 2 mg l^?1 NH₂Cl resulted in the same or an increased log density of viable M. mucogenicum in the biofilm while inactivating some of the Proteobacteria. Although M. mucogenicum preferentially resided in the biofilm, NH₂Cl exposure caused release of viable M. mucogenicum from the biofilm into the water. Additional studies with this model should determine if sodium hypochlorite has a comparative effect and if other nontuberculous mycobacteria (NTM) respond to NH₂Cl similarly.     

The effects of cleaning and disinfection in reducing Salmonella contamination in a laboratory model kitchen

AIMS: To establish a laboratory model to compare the effectiveness of detergent-based disinfection procedures for reducing cross-contamination risks during handling of contaminated chicken. METHODS AND RESULTS: During handling of chickens, artificially contaminated with Salmonella enteritidis PT4, the organism was widely spread to hands, cloths, and hand- and food-contact surfaces. Hygiene procedures were assessed on the basis of their ability to reduce the number of recoverable salmonellas to <1 CFU. Although detergent-based cleaning using a typical bowl-wash routine without rinsing produced some risk reduction (from 100 to 61.4% of contaminated surfaces), it was insufficient to consistently restore surfaces to a hygienic state. By combining detergent-based cleaning with a rinsing step or with hypochlorite at 500 ppm (of available chlorine) some further reduction in microbial risk was achieved, but was not considered satisfactory for food hygiene purposes. By contrast the risk reduction produced by hypochlorite at 5000 ppm was highly significant and was sufficient to reduce the number of contaminated surfaces to 2.9%. CONCLUSIONS: A key step in achieving a hygienic state through detergent-based cleaning is rinsing but even this will not produce a 'hygienic' result for difficult surfaces such as the chopping board or the dishcloth. Disinfectant compounds should be considered in order to reduce the potential for foodborne cross infection within the home environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Although tests are available to determine the performance of disinfectants, there are no quantitative procedures available to compare the risk reduction achieved by disinfection with that produced by detergent-based procedures. This study describes a reproducible laboratory method which can be used to differentiate the effectiveness of different hygiene procedures for reducing cross-contamination risks during food handling.     

Viability of a marine microbial biofilm

The formation of a microbial biofilm on glass surfaces arranged in lamellar piles parallel with circulating sea water (3 cm·sec^–1) was studied. The increase in dry weight, protein content, nucleotide content (ATP, ADP), and diatoms was followed over a period of 62 days. Dry weight and protein were estimates of the total biofilm development, whereas the nucleotide measurements revealed the viability of the biofilm and reflected the dynamics in the community structure.     

A prototype model structure for mixed microbial populations in homogeneous food products

An important factor which has not been included in many models in the field of predictive microbiology is the influence of a background of microflora in a food product. It is however generally known that the growth of a microorganism as a pure culture can be substantially different from its growth in a mixed culture, due to microbial interactions. Because of the importance of these interactions and the lack of suitable modeling techniques in the field of predictive microbiology to describe them, the potential of models in other research fields-namely ecology-to deal with interactions is explored in previous work of the authors. However, a model structure for microbial growth in food products cannot simply be copied from those elaborated in ecology. The structure of a predictive growth model is indeed typical, primarily due to the explicit modeling of a lag phase. The current paper proposes a prototype model structure for growth of mixed microbial populations in homogeneous food products. The model is able to describe a lag phase and reduces to a classical predictive growth model in the special case of single-species growth.     

Impact of initial biofilm growth on the anode impedance of microbial fuel cells

Electrochemical impedance spectroscopy (EIS) was used to study the behavior of a microbial fuel cell (MFC) during initial biofilm growth in an acetate-fed, two-chamber MFC system with ferricyanide in the cathode. EIS experiments were performed both on the full cell (between cathode and anode) as well as on individual electrodes. The Nyquist plots of the EIS data were fitted with an equivalent electrical circuit to estimate the contributions of various intrinsic resistances to the overall internal MFC impedance. During initial development of the anode biofilm, the anode polarization resistance was found to decrease by over 70% at open circuit and by over 45% at 27 microA/cm(2), and a simultaneous increase in power density by about 120% was observed. The exchange current density for the bio-electrochemical reaction on the anode was estimated to be in the range of 40-60 nA/cm(2) for an immature biofilm after 5 days of closed circuit operation, which increased to around 182 nA/cm(2) after more than 3 weeks of operation and stable performance in an identical parallel system. The polarization resistance of the anode was 30-40 times higher than that of the ferricyanide cathode for the conditions tested, even with an established biofilm. For a two-chamber MFC system with a Nafion 117 membrane and an inter-electrode spacing of 15 cm, the membrane and electrolyte solution dominate the ohmic resistance and contribute to over 95% of the MFC internal impedance. Detailed EIS analyses provide new insights into the dominant kinetic resistance of the anode bio-electrochemical reaction and its influence on the overall power output of the MFC system, even in the high internal resistance system used in this study. These results suggest that new strategies to address this kinetic constraint of the anode bio-electrochemical reactions are needed to complement the reduction of ohmic resistance in modern designs.     

Effect of antifoam agents and efficiency of cleaning procedures on the cross-flow filtration of microbial suspensions

Summary The influence of several biocompatible antifoam agents on the performance of ultrafiltration membranes for yeast cell concentration is described. Flux rates of water solutions and cell suspensions decreased in the presence of the antifoam agents. The anti-foam fouling effect was cumulative. Water was ineffective as a cleaning substance, while 0.1 NaOH and ethanol were used successfully for membrane washing.     

Effects of temperature variability on community structure in a natural microbial food web

Climate change research has demonstrated that changing temperatures will have an effect on community‐level dynamics by altering species survival rates, shifting species distributions, and ultimately, creating mismatches in community interactions. However, most of this work has focused on increasing temperature, and still little is known about how the variation in temperature extremes will affect community dynamics. We used the model aquatic community held within the leaves of the carnivorous plant, Sarracenia purpurea, to test how food web dynamics will be affected by high temperature variation. We tested the community response of the first (bacterial density), second (protist diversity and composition), and third trophic level (predator mortality), and measured community respiration. We collected early and late successional stage inquiline communities from S. purpurea from two North American and two European sites with similar average July temperature. We then created a common garden experiment in which replicates of these communities underwent either high or normal daily temperature variation, with the average temperature equal among treatments. We found an impact of temperature variation on the first two, but not on the third trophic level. For bacteria in the high‐variation treatment, density experienced an initial boost in growth but then decreased quickly through time. For protists in the high‐variation treatment, alpha‐diversity decreased faster than in the normal‐variation treatment, beta‐diversity increased only in the European sites, and protist community composition tended to diverge more in the late successional stage. The mortality of the predatory mosquito larvae was unaffected by temperature variation. Community respiration was lower in the high‐variation treatment, indicating a lower ecosystem functioning. Our results highlight clear impacts of temperature variation. A more mechanistic understanding of the effects that temperature, and especially temperature variation, will have on community dynamics is still greatly needed.     

Impact of anodophilic biofilm bioelectroactivity on the denitrification behavior of air-cathode microbial fuel cells

Generally, high bioelectroactivity of anodophilic biofilm favors high power generation of microbial fuel cell (MFC); however, it is not clear whether it can promote denitrification of MFC synchronously. In this study, we studied the impact of anodophilic biofilm bioelectroactivity on the denitrification behavior of air-cathode MFC (AC-MFC) in steady state and found that high bioelectroactivity of anodophilic biofilm not only favored high power generation of the AC-MFC, but also promoted the growth of denitrifers at the anodes and strengthened denitrification. Anodophilic biofilms of AC-MFC with various bioelectroactivity were acclimated at conditions of open circuit (OC), R_ext of 1000 Ω and 20 Ω (denoted as AC-MFC-OC, AC-MFC-1000Ω, and AC-MFC-20Ω, respectively) and performed for over 100 days. Electrochemical tests and microbial analysis results showed that the anode of the AC-MFC-20Ω delivered higher current response of both oxidation and denitrification and had higher abundance of electroactive bacteria than the AC-MFC-OC, AC-MFC-1000Ω, demonstrating a higher bioelectroactivity of the anodophilic biofilms. Moreover, these electroactive bacteria favored the accumulation of denitrifers, like Thauera and Alicycliphilus, probably by consuming trace oxygen through catalyzing oxygen reduction. The AC-MFC-20Ω not only delivered a 61.7% higher power than the AC-MFC-1000Ω, but also achieved a stable and high denitrification rate constant (k_DN) of 1.9 h^?1, which was 50% and 40% higher than that of the AC-MFC-OC and AC-MFC-1000Ω, respectively. It could be concluded that the high bioelectroactivity of the anodophilic biofilms not only favored high power generation of the AC-MFC, but also promoted the enrichment of denitrifers at the anodes and strengthened denitrification. This study provided an effective method for enhancing power generation and denitrification performance of the AC-MFC synchronously.     

A kinetic model for microbial growth on solid hydrocarbons

A kinetic model has been presented to explain the growth of microorganism on solid hydrocarbons. The model is based on the assumption that metabolite produced by the growing cells helps the dissolution of the solid substrate in the aqueous medium. The linear behavior of the growth curve predicted by the model is verified experimentally.     

Effect of magnetic hyperthermia on the structure of biofilm and cellular viability of a food spoilage bacterium

This work evaluated the effect of magnetic hyperthermia (MH) on planktonic cells and biofilms of a major food spoilage bacterium Pseudomonas fluorescens and its performance compared to a conventional direct heating (DH) technique. The results showed that MH had a greater and faster bactericidal effect, promoting a significant reduction in cell viability (≥3 Log CFU) in planktonic and biofilm cells, and leading to a complete eradication of planktonic cells at 55 °C (after only ~8 min). Accordingly, when comparing the same final temperatures, MH was more harmful to the integrity of cell membranes than DH, as observed in confocal laser scanning microscope images. Additionally, scanning electron microscope images revealed that exposure to MH had promoted modifications of the bacterial cell surface as well as of the structure of the biofilm. These results present the possibility of using MH out of the biomedical field as a potential disinfection method in food-related environments.     

Effect of bulk liquid BOD concentration on activity and microbial community structure of a nitrifying,membrane-aerated biofilm

Membrane-aerated biofilms (MABs) are an effective means to achieve nitrification and denitrification of wastewater. In this research, microsensors, fluorescence in situ hybridization (FISH), and modeling were used to assess the impact of bulk liquid biological oxygen demand (BOD) concentrations on the activity and microbial community structure of nitrifying MABs. With 1 g m⁻³ BOD in the bulk liquid, the nitrification rate was 1.3 g N m⁻² day⁻¹, slightly lower than the 1.5 g N m⁻² day⁻¹ reported for no bulk liquid BOD. With bulk liquid BOD concentrations of 3 and 10 g m⁻³, the rates decreased to 1 and 0.4 g N m⁻² day⁻¹, respectively. The percent denitrification increased from 20% to 100% when the BOD increased from 1 to 10 g m⁻³ BOD. FISH results indicated increasing abundance of heterotrophs with increasing bulk liquid BOD, consistent with the increased denitrification rates. Modeling was used to assess the effect of BOD on nitrification rates and to compare an MAB to a conventional biofilm. The model-predicted nitrification rates were consistent with the experimental results. Also, nitrification in the MAB was much less sensitive to BOD inhibition than the conventional biofilm. The MAB achieved concurrent nitrification and denitrification, whereas little denitrification occurred in the conventional biofilm.