Impact of Fe2+^{2+} and Shear Stress on the Development and Mesoscopic Structure of Biofilms—A Bacillus subtilis Case Study

Bivalent cations are known to affect the structural and mechanical properties of biofilms. In order to reveal the impact of Fe2+ ions within the cultivation medium on biofilm development, structure and stability, Bacillus subtilis biofilms were cultivated in mini-fluidic flow cells. Two different Fe2+ inflow concentrations (0.25 and 2.5 mg/L, respectively) and wall shear stress levels (0.05 and 0.27 Pa, respectively) were tested. Mesoscopic biofilm structure was determined daily in situ and non-invasively by means of optical coherence tomography. A set of ten structural parameters was used to quantify biofilm structure, its development and change. The study focused on characterizing biofilm structure and development at the mesoscale (mm-range). Therefore, biofilm replicates (n = 10) were cultivated and analyzed. Three hypotheses were defined in order to estimate the effect of Fe2+ inflow concentration and/or wall shear stress on biofilm development and structure, respectively. It was not the intention to investigate and describe the underlying mechanisms of iron incorporation as this would require a different set of tools applied at microscopic levels as well as the use of, i.e., omic approaches. Fe2+ addition influenced biofilm development (e.g., biofilm accumulation) and structure markedly. Experiments revealed the accumulation of FeO(OH) within the biofilm matrix and a positive correlation of Fe2+ inflow concentration and biofilm accumulation. In more detail, independent of the wall shear stress applied during cultivation, biofilms grew approximately four times thicker at 2.5 mg Fe2+/L (44.8 µmol/L; high inflow concentration) compared to the low Fe2+ inflow concentration of 0.25 mg Fe2+/L (4.48 µmol/L). This finding was statistically verified (Scheirer–Ray–Hare test, ANOVA) and hints at a higher stability of Bacillus subtilis biofilms (e.g., elevated cohesive and adhesive strength) when grown at elevated Fe2+ inflow concentrations

Removal of Diverse and Abundant ARGs by MF-NF Process from Pig Manure and Digestate

Antimicrobial resistances are emerging as one main threat to worldwide human health and are expected to kill 10 million people by 2050. Intensive livestock husbandry, along with biogas digestate, are considered as one of the biggest ARG reservoirs. Despite major concerns, little information is available on the diversity and abundance of various ARGs in small to large scale pig farms and biogas digestate slurry in Germany, followed by their consequent removal using microfiltration (MF)–nanofiltration (NF) process. Here, we report the identification and quantification of 189 ARGs in raw manure and digestate samples, out of which 66 ARGs were shared among manures and 53 ARGs were shared among both manure and digestate samples. The highest reported total ARG copy numbers in a single manure sampling site was 1.15 × 10$^8$ copies/100 µL. In addition, we found the absolute concentrations of 37 ARGs were above 10$^5$ copies/100 μL. Filtration results showed that the highly concentrated ARGs (except aminoglycoside resistance ARGs) in feed presented high log retention value (LRV) from 3 to as high as 5 after the MF-NF process. Additionally, LRV below 2 was noticed where the initial absolute ARG concentrations were ≤10$^3$ copies/100 μL. Therefore, ARG removal was found to be directly proportional to its initial concentration in the raw manure and in digestate samples. Consequently, some ARGs (tetH, strB) can still be found within the permeate of NF with up to 10$^4$ copies/100 μL

On-Line Monitoring of Biofilm Accumulation on Graphite-Polypropylene Electrode Material Using a Heat Transfer Sensor

Biofilms growing on electrodes are the heart piece of bioelectrochemical systems (BES). Moreover, the biofilm morphology is key for the efficient performance of BES and must be monitored and controlled for a stable operation. For the industrial use of BES (i.e., microbial fuel cells for energy production), monitoring of the biofilm accumulation directly on the electrodes during operation is desirable. In this study a commercially available on-line heat transfer biofilm sensor is applied to a graphite-polypropylene (C-PP) pipe and compared to its standard version where the sensor is applied to a stainless-steel pipe. The aim was to investigate the transferability of the sensor to a carbonaceous material (C-PP), that are preferably used as electrode materials for bioelectrochemical systems, thereby enabling biofilm monitoring directly on the electrode surface. The sensor signal was correlated to the gravimetrically determined biofilm thickness in order to identify the sensitivity of the sensor for the detection and quantification of biofilm on both materials. Results confirmed the transferability of the sensor to the C-PP material, despite the sensor sensitivity being decreased by a factor of approx. 5 compared to the default biofilm sensor applied to a stainless-steel pipe

Impact of Livestock Farming on Nitrogen Pollution and the Corresponding Energy Demand for Zero Liquid Discharge

Intensive livestock farming has negatively impacted the environment by contributing to the release of ammonia and nitrous oxide, groundwater nitrate pollution and eutrophication of rivers and estuaries. The nitrogen footprint calculator has predicted the large impact of meat production on global nitrogen loss, but it could not form the relationship between meat production and the corresponding manure generation. Here we report on the formation of direct relationships between beef, pork and poultry meat production and the corresponding amount of nitrogen loss through manure. Consequently, the energy demand for ammonium nitrogen recovery from manure is also reported. Nitrogen loss to the environment per unit of meat production was found directly proportional to the virtual nitrogen factors. The relationship between total nitrogen intake and the corresponding nitrogen loss per kg of meat production was also found linear. Average nitrogen loss due to manure application was calculated at 110 g kg$^{−1}$ for poultry. The average nitrogen loss increased to 190 and 370 g-N kg$^{−1}$ for pork and beef productions, respectively. Additionally, 147 kg ammonium nitrogen was calculated to be recovered from 123 m$^{3}$ of manure. This corresponded to 1 Mg of beef production. The recovery of ammonium nitrogen was reduced to 126 and 52 kg from 45 and 13 m3 of pork and poultry manure, respectively. The ammonium nitrogen recovery values were calculated with respect to 1 Mg of both pork and poultry meat productions. Consequently, the specific energy demand of ammonium nitrogen recovery from beef manure was noticed at 49 kWh kg$^{−1}$, which was significantly 57% and 69% higher than that of pork and poultry manure, respectively

Removal of microplastics from waters through agglomeration-fixation using organosilanes—effects of polymer types, water composition and temperature

Due to the fact, that microplastics are a global environmental problem, new ways for their removal from water, soil and air need to be developed. New materials in combination with easy to implement technologies for microplastic removal come into the focus of scientific studies and engineering, especially for application in water resources. In our comparative case study, the effects of water composition and temperature on the agglomeration-fixation reaction of microplastics using organosilanes were examined. We compared biologically treated municipal wastewater, seawater and demineralized water at temperatures ranging from 7.5–40 °C. Temperature variations and tested water compositions showed no negative effect on microplastic removal. The residues of the organosilanes remaining in the water after the fixation process were monitored using ICP-OES and DOC measurements. Only one of the organosilanes tested showed no dissolved residues in the waters. Microplastic encompasses a multitude of different types of polymers with different properties and surface chemistries. Therefore, we compared the efficiency of the process for polyethylene, polypropylene, polyamide, polyester, and polyvinylchloride as examples of common polymer types with different chemical compositions. A strong effect of the polarity of microplastics and organosilanes on removal efficiency was observed. The organic groups of organosilanes can be chemically adapted to different polymer types

Impact of log(Kow) Value on the Extraction of Antibiotics from River Sediments with Pressurized Liquid Extraction

The quantification of antibiotics (ABs) in sediments is an analytical challenge, but at the same time, it is indispensable to understand the fate of ABs in aquatic systems such as rivers. The aim of this study was to develop a comprehensive method to determine 19 ABs classified as macrolides, sulfonamides, fluoroquinolones, tetracyclines, clindamycin and trimethoprim in river sediments, using a combination of pressurized liquid extraction and solid phase extraction with the separation and detection with liquid chromatography coupled with mass spectrometry. Our results showed that the physical-chemical properties (e.g., log(Kow) value) of the analytes affected the extraction efficiency. Therefore, we propose to order ABs based on their log(Kow) values instead of traditional classification (macrolides, sulfonamides etc.) to select a suitable extraction solvent. ABs with log(Kow) values below zero (mainly fluoroquinolones and tetracyclines) were difficult to extract with all of the tested protocols compared to ABs with a log(Kow) larger than zero. After comparing different extraction protocols for ABs from solid and sediments, we concluded that recoveries in the range of 0.8 to 64.8% could be achieved for ABs with a log(Kow) value larger than zero using a mixture of acetonitrile and 50 mM phosphoric acid (50/50, v/v) in two extraction cycles at 100 °C

Improving the Cathodic Biofilm Growth Capabilities of Kyrpidia spormannii EA-1 by Undirected Mutagenesis

The biotechnological usage of carbon dioxide has become a relevant aim for future processes. Microbial electrosynthesis is a rather new technique to energize biological CO$_{2}$ fixation with the advantage to establish a continuous process based on a cathodic biofilm that is supplied with renewable electrical energy as electron and energy source. In this study, the recently characterized cathodic biofilm forming microorganism Kyrpidia spormannii strain EA-1 was used in an adaptive laboratory evolution experiment to enhance its cathodic biofilm growth capabilities. At the end of the experiment, the adapted cathodic population exhibited an up to fourfold higher biofilm accumulation rate, as well as faster substratum coverage and a more uniform biofilm morphology compared to the progenitor strain. Genomic variant analysis revealed a genomically heterogeneous population with genetic variations occurring to various extends throughout the community. Via the conducted analysis we identified possible targets for future genetic engineering with the aim to further optimize cathodic growth. Moreover, the results assist in elucidating the underlying processes that enable cathodic biofilm formation

Desalination of Seawater Using Cationic Poly(acrylamide) Hydrogels and Mechanical Forces for Separation

In this study, the ability of cationic poly(acrylamide-co-(3-acrylamidopropyl)trimethylammonium chloride) hydrogels to desalinate seawater is explored, where the salt separation is based on the partial rejection of mobile salt ions by the fixed charges along the polymer backbone. Water absorbency measurements reveal that artificial seawater-containing divalent ions (Mg$^{2+}$, Ca$^{2+}$, and SO$_{4}$$^{2-}$) drastically decrease the swelling capacity of previously employed anionic poly(acrylic acid-co-sodium acrylate) hydrogels, whereas no influence on the swelling behavior of the synthesized cationic hydrogels is found. The swelling behavior and mechanical properties are studied by varying the degree of crosslinking and degree of ionization systematically in the range of 1–5 and 25–75 mol%, respectively. Finally, artificial seawater (c$_{sea}$= 0.171 mol L$^{-1}$) is desalinated in a custom-built press setup with an estimated efficiency of E$_{m³}$= 17.6 kWh m$^{-3}$ by applying an external pressure on the swollen hydrogels

Investigation of biofilm growth within a monodisperse porous medium under fluctuating water level assessed by means of mri

Microorganisms settle in diverse partially saturated porous media in the form of biofilms. The alteration of hydraulic properties and diffusive transport processes occurs simultaneously with biofilm growth in porous media. Imaging methods offer the ability to directly visualize and quantify alterations on the pore scale. However, imaging methods have mainly observed biofilm growth in completely saturated porous media. The current study used magnetic resonance imaging (MRI) to dynamically visualize biofilm growth within a porous medium under alternating drainage and flushing events. Prior to the MRI experiments, the sample was cultivated for 6 days within a porous medium consisting of 2 mm glass spheres. Starting from day 6, growth was monitored using MRI over a period of 7 days. The approach allowed for a visualization of all fractions (biofilm, water, air, and porous material) after drainage as well as flushing events. Biofilm was found to preferentially grow within permanently wetted areas situated next to pore throats. Furthermore, an increase in the water retention and connectivity of the liquid phase was found. The largest liquid cluster covered 11% (day 6) and 91% (day 12) of the total retained water, suggesting that biofilm growth might improve diffusive transport processes within partially saturated porous medi

MF–NF Treatment Train for Pig Manure: Nutrient Recovery and Reuse of Product Water

The livestock industry negatively impacts the environment by producing high organic and mineral loaded manure and wastewater. On the contrary, manure is also considered as the major focal point of resource recovery. The microfiltration (MF) process in manure treatment is well known for being the least complex and highly energy efficient. However, the major fraction of the dissolve nutrients easily bypasses the MF membranes. In this research work, we reported the efficiency of using MF–nanofiltration (NF) treatment train in a dead-end filtration system for the treatment of raw manure. The objectives were to produce nutrient rich separate streams in reduced volumes and a particle and pathogen-free product water. MF removed TSS above 98% and the COD and phosphorus (P) retention were noticed above 60 and 80%, respectively, within a reduced MF concentrate volume, which accounted for 40% of the initial feed volume. The NF of MF permeate by NF270 showed most promising results by concentrating overall 50 and 70% of the total nitrogen (TN) and potassium (K) within a reduced NF concentrate volume, which accounted for 30% of the initial MF feed volume. Finally, the MF–NF treatment train of raw pig manure could produce a particle-free product water that can be reused in farms to wash barns, to irrigate nearby cultures, or can be applied to specific fields based on the demand