Investigating alternative materials to EPDM for automatic taps in the context of Pseudomonas aeruginosa and biofilm control.

BACKGROUND: Automatic taps use solenoid valves (SVs) which incorporate a rubber (typically EPDM) diaphragm to control water flow. Contaminated SVs can be reservoirs of opportunistic pathogens such as Pseudomonas aeruginosa; an important cause of healthcare-associated infection. AIMS: To investigate the attachment and biofilm formation of P. aeruginosa on EPDM and relevant alternative rubbers to assess the impact on water hygiene in a laboratory model. METHODS: Biofilm formation on EPDM, silicone and nitrile rubber coupons was investigated using a CDC biofilm reactor. SVs incorporating EPDM or nitrile rubber diaphragms were installed on to an experimental water distribution system (EWDS) and inoculated with P. aeruginosa. P. aeruginosa water levels were monitored for 12-weeks. SVs incorporating diaphragms (EPDM, silicone or silver ion-impregnated silicone rubber), pre-colonized with P. aeruginosa, were installed and the effect of flushing as a control measure was investigated. The concentration of P. aeruginosa in the water was assessed by culture and biofilm assessed by culture and microscopy. FINDINGS: Bacterial attachment was significantly higher on nitrile (6.2 × 105 cfu/coupon) and silicone (5.4 × 105 cfu/coupon) rubber than on EPDM (2.9 ×105 cfu/coupon) (P<0.05, N = 17). Results obtained in vitro did not translate to the EWDS where, after 12-weeks in situ, there was no significant difference in P. aeruginosa water levels or biofilm levels. Flushing caused a superficial reduction in bacterial counts after <5 min of stagnation. CONCLUSION: This study did not provide evidence to support replacement of EPDM with (currently available) alternative rubbers and indicated the first sample of water dispensed from a tap should be avoided for use in healthcare settings.

Synthesis and characterization of antibacterial polylactic acid film incorporated with cinnamaldehyde inclusions for fruit packaging.

To maintain the quality of postharvest fruits continuously and meet the health requirements of consumers, a high barrier and long-lasting antibacterial polylactic acid film as packaging material was developed in this study. Polylactic acid-based antibacterial films incorporated with Cinnamaldehyde inclusions were used to achieve long-lasting antibacterial activity and improve the barrier properties. Cinnamaldehyde inclusions were prepared via the inclusion method and used as a sustained-release antibacterial agent and reinforcement to be incorporated into polylactic acid-based films within a concentration range of 0-30 wt%. The FT-IR spectrum demonstrated that the Cinnamaldehyde inclusions was physically interacting with PLA. The XRD results showed that the cinnamaldehyde inclusions at a concentration of 10 wt% enhanced the crystallinity of the antibacterial film. The oxygen and water vapor barrier properties of the film were respectively 14.29% and 12.38% higher than those of a pure PLA film. The tensile strength of the antibacterial film increased by 20%. And the antibacterial activity against Escherichia coli and Listeria monocytogenes was 100%. The release rate of cinnamaldehyde of the antibacterial film was slow and varied smoothly for 20 d.

Pectinolytic Bacterial Consortia Reduce Jute Retting Period and Improve Fibre Quality.

Jute fibre is the second most important fibre next to cotton. It is obtained from the bark of plant through microbial retting process. Here we report optimized microbial retting protocol that can lower retting period and produce high fibre quality. A total of 451 bacterial colonies have been isolated from five jute retting water samples in Bangladesh. Higher pectinolytic bacterial isolates were predominant in the later stage of jute retting. Out of these, 168 isolates have been screened by both semi-quantitative and quantitative pectinase, xylanase and cellulase enzyme assay. Among them, 144 isolates have been selected on the basis of extra cellular enzyme activity of these three enzymes. 16 s ribosomal gene sequencing analysis identified 2 phyla- Firmicutis (80.55%) and Proteobacteria (19.45%). To check the synergistic and antagonistic effect 10 selected isolates were tested in 167 different combinations. Three best combinations were identified that lowered retting period from 18-21 days to 10 days producing high quality fibre in both laboratory and field trial. This improved retting technology can be adopted in industrial scale for the production of quality jute fibre in a controlled condition in reduced water quantity without polluting the environment.

Marine biofilm bacterial community response and carbon steel loss following Deepwater Horizon spill contaminant exposure.

Steel marine structures provide foci of biodiversity when they develop into artificial reefs. Development begins with deposition of a biofilm. The effects of contaminants from oil spills on biofilm microbiomes, microbially-induced corrosion (MIC) and metal loss may impact preservation of marine metal structures. A microcosm experiment exposed biofilms on carbon steel disks (CSDs) to crude oil, dispersant, and dispersed oil to address their impacts on bacterial composition and metal loss and pitting. Biofilm diversity increased over time in all exposures. Community composition in dispersant and dispersed oil treatments deviated from the controls for the duration of a 12-week experiment. As biofilms matured, Pseudomonadaceae increased while Rhodobacteraceae decreased in abundance in dispersed oil treatments compared to the controls and dispersant treatments. Greatest mass loss and deepest pitting on CSDs were observed in dispersed oil treatments, suggesting impacts manifest as a consequence of increased MIC potential on carbon steel.

Multispecies biofilm formation by the contaminating microbiota in raw milk.

Biofilms can be formed on the surfaces of dairy processing equipment and are a potential source of product contamination. This study evaluated the diversity of multispecies biofilms formed on stainless steel (SS) due to the contaminating microbiota in raw milk. Samples of raw milk were used: one was fresh milk and the other maintained in refrigerated bulk tanks for up to 48 h. The mesophilic aerobic contamination was ∼104 CFU ml-1 in fresh milk and 106 CFU ml-1 in bulk milk. SS coupons were kept immersed in the milk at 7 ±2 °C for 10 days, and every two days, the raw milk was changed for samples of the same origin collected on the current day. After incubation for 10 days, sessile cells in the biofilm reached 105 CFU cm-2 in the presence of fresh milk, and 106 CFU cm-2 in the presence of bulk milk. The genetic diversity analysis showed that Gammaproteobacteria and Bacilli predominated in the biofilms throughout the incubation of both milk samples and these biofilms showed a reduction in diversity over time. The main classes of bacteria found in these biofilms have representatives of great importance since many of them have spoilage potential.

The corrosion process caused by the activity of the anaerobic sporulated bacterium Clostridium celerecrescens on API XL 52 steel.

The microbial corrosion of oil and gas pipes is one of the problems occurring in the oil industry. Various mechanisms explaining microbial corrosion have been demonstrated. Commonly, biocorrosion is attributed to sulfate-reducing bacteria. Also, it has recently been reported that microbial species can connect their electron transport system to metal electrodes. In this research, two spore-forming bacteria isolated in different years from a gas pipeline were identified by biochemical techniques and by 16S rDNA amplification, sequencing, and comparison with the NCBI database. Isolates were also compared between them using molecular techniques as the restriction patterns, unique for 16S rDNA (ARDRA), and the profile of the amplified bit from the genomic DNA, using an unspecific primer (RAPD). The results obtained showed that both isolates corresponded to Clostridium celerecrescens with a 99% similarity according to the sequence reported on the NCBI database. Also, the ARDRA and RAPD electrophoretic profiles of both strains were identical, and no plasmids were found in the strains. Thus, it can be settled that this bacterium is persistent in the environment prevailing in gas pipelines. Also, it was demonstrated that the bacterial secretion of organic acids contributes to the pitting and general biocorrosion of API XL 52 steel. The rates of corrosion obtained, approximately after 40 days, were correlated with the presence and metabolic activity of C. celerecrescens on the metallic surfaces.

Estimating Bacterial Concentrations in Fibrous Substrates Through a Combination of Scanning Electron Microscopy and ImageJ.

Conventional signal-based microanalytical techniques for estimating bacterial concentrations are often susceptible to false signals. A visual quantification, therefore, may compliment such techniques by providing additional information and support better management decisions in the event of outbreaks. Herein, we explore a method that combines electron microscopy (EM) and image-analysis techniques and allows both visualization and quantification of pathogenic bacteria adherent even to complex nonuniform substrates. Both the estimation and imaging parameters were optimized to reduce the estimation error ( E, %) to close to ±5%. The method was validated against conventional microbiological techniques such as the use of optical density, flow cytometry, and quantitative real-time PCR (qPCR). It could easily be tailored to estimate different species of pathogens, such as Escherichia coli O157, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis, and Bacillus anthracis, on samples similar to those in real-time contamination scenarios. The present method is sensitive enough to detect ∼100 bacterial CFU/mL but has the potential to estimate even lower concentrations with increased imaging and computation times. Overall, this imaging-based method may greatly complement any signal-based pathogen-detection technique, especially in negating false signals, and therefore may significantly contribute to the field of analytical microbiology and biochemistry.

Microorganisms @ materials surfaces in aircraft: Potential risks for public health? - A systematic review.

BACKGROUND: Civil air travel is increasingly recognized as an important potential source for the rapid spread of infectious diseases that were geographically confined in the past, creating international epidemics with great health and socio-economic impact. OBJECTIVE: The objective of this systematic review is to elucidate the correlations of materials surfaces (composition, structure, properties) and microbial dependences on them in aircraft. METHODS: The review was prepared according to PRISMA guidelines. Based on a systematic search for studies published before 30 June 2018 in English, we selected and reviewed the contamination, tenacity, and transmission of microorganisms related to specific surfaces within the aircraft cabin. We also reviewed the chemical composition and properties of these surface materials applied within aircraft. RESULTS: From a total of 828 records 15 articles were included for further analysis in this systematic review, indicating that the aircraft interior surfaces in seat areas (tray tables, armrests, seat covers) and lavatories (door knob handles, toilet flush buttons) are generally colonized by various types of potentially hazardous microorganisms. CONCLUSIONS: The interior surfaces in seat and lavatory areas could pose higher health risks by causing infections due to their relatively high microbial contamination compared with other interior surfaces. The classification, chemical composition, surface structures and physicochemical properties of materials surfaces have a varied effect on the adhesion, colonization, tenacity and potential transmission of microorganisms within the aircraft cabin. Strategies are proposed for the interruption of surface-related infection chains in the aircraft field.

Microbial network of the carbonate precipitation process induced by microbial consortia and the potential application to crack healing in concrete.

Current studies have employed various pure-cultures for improving concrete durability based on microbially induced carbonate precipitation (MICP). However, there have been very few reports concerned with microbial consortia, which could perform more complex tasks and be more robust in their resistance to environmental fluctuations. In this study, we constructed three microbial consortia that are capable of MICP under aerobic (AE), anaerobic (AN) and facultative anaerobic (FA) conditions. The results showed that AE consortia showed more positive effects on inorganic carbon conversion than AN and FA consortia. Pyrosequencing analysis showed that clear distinctions appeared in the community structure between different microbial consortia systems. Further investigation on microbial community networks revealed that the species in the three microbial consortia built thorough energetic and metabolic interaction networks regarding MICP, nitrate-reduction, bacterial endospores and fermentation communities. Crack-healing experiments showed that the selected cracks of the three consortia-based concrete specimens were almost completely healed in 28 days, which was consistent with the studies using pure cultures. Although the economic advantage might not be clear yet, this study highlights the potential implementation of microbial consortia on crack healing in concrete.

Microbial fouling and corrosion of carbon steel in deep anoxic alkaline groundwater.

Understanding the corrosion of carbon steel materials of low and intermediate level radioactive waste under repository conditions is crucial to ensure the safe storage of radioactive contaminated materials. The waste will be in contact with the concrete of repository silos and storage containers, and eventually with groundwater. In this study, the corrosion of carbon steel under repository conditions as well as the microbial community forming biofilm on the carbon steel samples, consisting of bacteria, archaea, and fungi, was studied over a period of three years in a groundwater environment with and without inserted concrete. The number of biofilm forming bacteria and archaea was 1,000-fold lower, with corrosion rates 620-times lower in the presence of concrete compared to the natural groundwater environment. However, localized corrosion was detected in the concrete-groundwater environment indicating the presence of local microenvironments where the conditions for pitting corrosion were favorable.

Bioprinting: an assessment based on manufacturing readiness levels.

Over the last decade, bioprinting has emerged as a promising technology in the fields of tissue engineering and regenerative medicine. With recent advances in additive manufacturing, bioprinting is poised to provide patient-specific therapies and new approaches for tissue and organ studies, drug discoveries and even food manufacturing. Manufacturing Readiness Level (MRL) is a method that has been applied to assess manufacturing maturity and to identify risks and gaps in technology-manufacturing transitions. Technology Readiness Level (TRL) is used to evaluate the maturity of a technology. This paper reviews recent advances in bioprinting following the MRL scheme and addresses corresponding MRL levels of engineering challenges and gaps associated with the translation of bioprinting from lab-bench experiments to ultimate full-scale manufacturing of tissues and organs. According to our step-by-step TRL and MRL assessment, after years of rigorous investigation by the biotechnology community, bioprinting is on the cusp of entering the translational phase where laboratory research practices can be scaled up into manufacturing products specifically designed for individual patients.

Biomachining: metal etching via microorganisms.

The use of microorganisms to remove metal from a workpiece is known as biological machining or biomachining, and it has gained in both importance and scientific relevance over the past decade. Conversely to mechanical methods, the use of readily available microorganisms is low-energy consuming, and no thermal damage is caused during biomachining. The performance of this sustainable process is assessed by the material removal rate, and certain parameters have to be controlled for manufacturing the machined part with the desired surface finish. Although the variety of microorganisms is scarce, cell concentration or density plays an important role in the process. There is a need to control the temperature to maintain microorganism activity at its optimum, and a suitable shaking rate provides an efficient contact between the workpiece and the biological medium. The system's tolerance to the sharp changes in pH is quite limited, and in many cases, an acid medium has to be maintained for effective performance. This process is highly dependent on the type of metal being removed. Consequently, the operating parameters need to be determined on a case-by-case basis. The biomachining time is another variable with a direct impact on the removal rate. This biological technique can be used for machining simple and complex shapes, such as series of linear, circular, and square micropatterns on different metal surfaces. The optimal biomachining process should be fast enough to ensure high production, a smooth and homogenous surface finish and, in sum, a high-quality piece. As a result of the high global demand for micro-components, biomachining provides an effective and sustainable alternative. However, its industrial-scale implementation is still pending.

Molecular Analysis of Bacterial Microbiota on Brazilian Currency Note Surfaces.

Currency notes have been implicated as a vehicle for transmitting community-acquired bacterial infections. However, the overall diversity of the bacterial population residing on banknotes is still unknown in Brazil. In this study, we aimed to investigate the overall bacterial population from 150 different Brazilian Rial (R$) notes in circulation using a culture-independent Illumina massively parallel sequencing approach of the 16S rRNA genes. Samples were randomly collected from three different street markets or "feiras" in the metropolitan region of São Paulo. Taxonomical composition revealed the abundance of Proteobacteria phyla, followed by Firmicutes and Streptophyta, with a total of 1193 bacterial families and 3310 bacterial genera. Most of these bacterial genera are of human, animal, and environmental origins. Also, our analysis revealed the presence of some potential pathogenic bacterial genera including Salmonella, Staphylococcus, and Klebsiella. The results demonstrate that there is a tremendous diversity of bacterial contamination on currency notes, including organisms known to be opportunistic pathogens. One of the factors that may contribute to the richness of bacterial diversity in currency notes is personal hygiene. Thus, our results underscore the need to increase public awareness of the importance of personal hygiene of money handlers who also handle food.

Corrosion behaviour and biocorrosion of galvanized steel water distribution systems.

Galvanized steel tubes are a popular mean for water distribution systems but suffer from corrosion despite their zinc or zinc alloy coatings. First, the quality of hot-dip galvanized (HDG) coatings was studied. Their microstructure, defects, and common types of corrosion were observed. It was shown that many manufactured tubes do not reach European standard (NBN EN 10240), which is the cause of several corrosion problems. The average thickness of zinc layer was found at 41µm against 55µm prescribed by the European standard. However, lack of quality, together with the usual corrosion types known for HDG steel tubes was not sufficient to explain the high corrosion rate (reaching 20µm per year versus 10µm/y for common corrosion types). Electrochemical tests were also performed to understand the corrosion behaviours occurring in galvanized steel tubes. Results have shown that the limiting step was oxygen diffusion, favouring the growth of anaerobic bacteria in steel tubes. EDS analysis was carried out on corroded coatings and has shown the presence of sulphur inside deposits, suggesting the likely bacterial activity. Therefore biocorrosion effects have been investigated. Actually sulphate reducing bacteria (SRB) can reduce sulphate contained in water to hydrogen sulphide (H2S), causing the formation of metal sulphides. Although microbial corrosion is well-known in sea water, it is less investigated in supply water. Thus, an experimental water main was kept in operation for 6months. SRB were detected by BART tests in the test water main.

Antifouling strategies and corrosion control in cooling circuits.

Biofouling and corrosion phenomena dramatically reduce the functionality of industrial cooling circuits, especially in marine environments. This study underlines the effectiveness of a low level chlorination treatment of seawater to prevent biological fouling and biocorrosion. Reported examples emphasize the reaction of chlorine with bromide, ammonia and organic compounds in seawater and the effectiveness of a treatment performed in such a way to guarantee a residual concentration lower than 3µM at the outlet of the condensers. In a brief review of antifouling strategies, alternatives to chlorination and the monitoring approach able to optimize the treatments are also reported. An integrated, on-line system based on electrochemical probes (Biox system and a linear polarization resistance probe) demonstrated to be sufficient to monitor in real time: corrosion, biofilm growth and chemical treatments based on chlorine or alternative oxidant products (chlorine dioxide, etc.). A careful electrochemical monitoring and the optimized treatments help the plant operators of industrial cooling circuits prevent the decay of the equipment performance, allowing at the same time the control of the halogenated by-products formation.

The study of marine corrosion of copper alloys in chlorinated condenser cooling circuits: the role of microbiological components.

The present paper reports the on-line monitoring of corrosion behavior of the CuNi 70:30 and Al brass alloys exposed to seawater and complementary offline microbiological analyses. An electrochemical equipment with sensors specifically set for industrial application and suitable to estimate the corrosion (by linear polarization resistance technique), the biofilm growth (by the BIOX electrochemical probe), the chlorination treatment and other physical-chemical parameters of the water has been used for the on-line monitoring. In order to identify and better characterize the bacteria community present on copper alloys, tube samples were collected after a long period (1year) and short period (2days) of exposition to treated natural seawater (TNSW) and natural seawater (NSW). From the collected samples, molecular techniques such as DNA extraction, polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE) and identification by sequencing were performed to better characterize and identify the microbial biodiversity present in the samples. The monitoring data confirmed the significant role played by biofouling deposition against the passivity of these Cu alloys in seawater and the positive influence of antifouling treatments based on low level dosages. Molecular analysis indicated biodiversity with the presence of Marinobacter, Alteromonas and Pseudomonas species.

Influence of respiratory substrate in carbon steel corrosion by a Sulphate Reducing Prokaryote model organism.

Sulphate Reducing Prokaryotes (SRP) are an important group of microorganisms involved in biocorrosion processes. Sulphide production is recognized as a fundamental cause of corrosion and nitrate is often used as treatment. The present work analyses the influence of respiratory substrates in the metal, from off-shore installations, SRP influenced corrosion, using Desulfovibrio desulfuricans ATTC 27774 as model organism, since this can switch from sulphate to nitrate. Open Circuit Potential over 6days in different conditions was measured, showing an increase around 200 and 90mV for the different media. Tafel plots were constructed allowing Ecorr and jcorr calculations. For SRP in sulphate and nitrate media Ecorr values of -824 and -728mV, and jcorr values of 2.5 and 3.7µAcm(-2), respectively, were attained indicating that in nitrate, the resultant corrosion rate is larger than in sulphate. Also, it is shown that the equilibrium of sulphide in the solution/gas phases is a key factor to the evolution of corrosion Nitrate prevents pitting but promotes general corrosion and increases the corrosion potential and iron dissolution 40 times when compared to sulphate. Our results demonstrate that nitrate injection strategy in oil fields has to be considered carefully as option to reduce souring and localized corrosion.

Impact of Desulfovibrio alaskensis biofilms on corrosion behaviour of carbon steel in marine environment.

Sulfate reducing prokaryotes are associated with the steel deterioration. They build heterogeneous biofilms, capable of accelerating corrosion processes. In this study metabolic activity and the biofilm development of Desulfovibrio alaskensis were correlated to electrochemical response of carbon steel surface. In the exponential growth phase sulfide concentration reached its maximum of about 10mM. This phenomenon was responsible for the parallel increase in the corrosion potential (Ecorr) up to -720mV (vs. SCE). Subsequently, during the intensive biofilm formation and development another Ecorr peak (-710mV vs. SCE) occurred. Decrease in Ecorr was registered during the biofilm maturation and kept stable, being 20mV lower than in the control. While carbon steel was protected from the microbial attachment and exposed to metabolic products, only one potential maximum (-730mV vs. SCE) was recorded. Here Ecorr variations coincided with sulfide concentration changes and kept at 120mV lower vs. the control. Weight loss examinations revealed corrosion rates, which did not exceed 0.05mm/y. Confocal microscopy suggested the importance of extracellular proteins in the biofilm formation. Above 150 proteins were detected in the EPS matrix. Surface effects of biofilm and metabolic products were visualised, revealing the role of attached microorganisms in the localised corrosion.

Combined geochemical and electrochemical methodology to quantify corrosion of carbon steel by bacterial activity.

The availability of respiratory substrates, such as H2 and Fe(II,III) solid corrosion products within nuclear waste repository, will sustain the activities of hydrogen-oxidizing bacteria (HOB) and iron-reducing bacteria (IRB). This may have a direct effect on the rate of carbon steel corrosion. This study investigates the effects of Shewanella oneidensis (an HOB and IRB model organism) on the corrosion rate by looking at carbon steel dissolution in the presence of H2 as the sole electron donor. Bacterial effect is evaluated by means of geochemical and electrochemical techniques. Both showed that the corrosion rate is enhanced by a factor of 2-3 in the presence of bacteria. The geochemical experiments indicated that the composition and crystallinity of the solid corrosion products (magnetite and vivianite) are modified by bacteria. Moreover, the electrochemical experiments evidenced that the bacterial activity can be stimulated when H2 is generated in a small confinement volume. In this case, a higher corrosion rate and mineralization (vivianite) on the carbon steel surface were observed. The results suggest that the mechanism likely to influence the corrosion rate is the bioreduction of Fe(III) from magnetite coupled to the H2 oxidation.

Influence of hydrogen-oxidizing bacteria on the corrosion of low carbon steel: Local electrochemical investigations.

Low carbon steel has been considered a suitable material for component of the multi-barrier system employed on the geological disposal of high-level radioactive waste (HLW). A non negligible amount of dihydrogen (H2) is expected to be produced over the years within the geological repository due to the anoxic corrosion of metallic materials and also to the water radiolysis. The influence of the activity of hydrogen-oxidizing bacteria (HOB) and iron-reducing bacteria (IRB) on carbon steel corrosion is considered in this study because of the high availability of energetic nutriments (H2, iron oxides and hydroxides) produced in anoxic disposal conditions. Local electrochemical techniques were used for investigating the activity of IRB as a promoter of local corrosion in the presence of H2 as electron donor. A local consumption of H2 by the bacteria has been evidenced and impedance measurements indicate the formation of a thick layer of corrosion products.