Colonization and spreading dynamics of Lactiplantibacillus plantarum spoilage isolates on wet surfaces.

The role of lactic acid bacteria, including Lactiplantibacillus plantarum, in food spoilage is well recognized, while the behavior of these non-motile bacteria on wet surfaces, such as those encountered in food processing environments has gained relatively little attention. Here, we observed a fast colony spreading of non-motile L. plantarum spoilage isolates on wet surfaces via passive sliding using solid BHI agar media as a model. We investigated the effect of physical properties of agar hydrogel substrate on the surface spreading of six L. plantarum food isolates FBR1-6 and a model strain WCFS1, using increasing concentrations of agar from 0.25 up to 1.5% (w/v). Our results revealed that L. plantarum strain FBR2 spreads significantly on low agar concentration plates compared to the other strains studied here (with a factor of 50-60 folds higher surface coverage), due to the formation of very soft biofilms with high water content that can float on the surface. The fast-spreading of FBR2 colonies is accompanied by an increased number of cells, elongated cell morphology, and a higher amount of extracellular components. Our finding highlights colonization dynamics and the spreading capacity of non-motile bacteria on surfaces that are relatively wet, thereby revealing an additional hitherto unnoticed parameter for non-motile bacteria that may contribute to contamination of foods by fast surface spreading of these bacteria in food processing environments.

Cation exchange membrane behaviour of extracellular polymeric substances (EPS) in salt adapted granular sludge.

This paper aims to elucidate the role of extracellular polymeric substances (EPS) in regulating anion and cation concentrations and toxicity towards microorganisms in anaerobic granular sludges adapted to low (0.22 M of Na+) and high salinity (0.87 M of Na+). The ion exchange properties of EPS were studied with a novel approach, where EPS were entangled with an inert binder (PVDF-HFP) to form a membrane and characterized in an electrodialysis cell. With a mixture of NaCl and KCl salts the EPS membrane was shown to act as a cation exchange membrane (CEM) with a current efficiency of ∼80%, meaning that EPS do not behave as ideal CEM. Surprisingly, the membrane had selectivity for transport of K+ compared to Na+ with a separation factor ( [Formula: see text] ) of 1.3. These properties were compared to a layer prepared from a model compound of EPS (alginate) and a commercial CEM. The alginate layer had a similar current efficiency (∼80%.), but even higher [Formula: see text] of 1.9, while the commercial CEM did not show selectivity towards K+ or Na+, but exhibited the highest current efficiency of 92%. The selectivity of EPS and alginate towards K+ transport has interesting potential applications for ion separation from water streams and should be further investigated. The anion repelling and cation binding properties of EPS in hydrated and dehydrated granules were further confirmed with microscopy (SEM-EDX, epifluorescence) and ion chromatography (ICP-OES, IC) techniques. Results of specific methanogenic activity (SMA) tests conducted with 0.22 and 0.87 M Na+ adapted granular sludges and with various monovalent salts suggested that ions which are preferentially transported by EPS are also more toxic towards methanogenic cells.

Biological treatment of produced water coupled with recovery of neutral lipids.

Produced water (PW) is the largest waste stream generated by oil and gas industry. It is commonly treated by physical-chemical processes due to high salt content and poor biodegradability of water insoluble compounds, such as n-alkanes. N-alkanes can represent a major fraction of organic contaminants within PW. In this study the possibility of simultaneous n - alkane biodegradation and production of neutral lipids in a concentrated PW stream with A. borkumenis SK2 as the sole reactor inoculum was investigated. N-alkane removal efficiency up to 99.6%, with influent alkane COD of 7.4 g/L, was achieved in a continuously operated reactor system. Gas chromatography results also showed that the majority of other non-polar compounds present in the PW were biodegraded. Biodegradation of n-alkanes was accompanied by simultaneous production of neutral lipids, mostly wax ester (WE)-alike compounds. We demonstrate, that under nutrient limited conditions and 108.9 ±â€¯3.3 mg/L residual n-alkane concentration the accumulation of extracellular WE-alike compounds can be up to 12 times higher compared to intracellular, reaching 3.08 grams per litre of reactor volume (g/Lreactor) extracellularly and 0.28 g/Lreactor intracellularly. With residual n-alkane concentration of 311.5 ±â€¯34.2 mg/L accumulation of extracellular and intracellular WE-alike compounds can reach up to 6.15 and 0.91 g/Lreactor, respectively. To the best of our knowledge simultaneous PW treatment coupled with production of neutral lipids has never been demonstrated before.

The potential of osmolytes and their precursors to alleviate osmotic stress of anaerobic granular sludge.

Increasing amounts of saline (waste)water with high concentrations of organic pollutants are generated globally. In the anaerobic (waste)water treatment domain, high salt concentrations are repeatedly reported to inhibit methanogenic activity and strategies to overcome this toxicity are needed. Current research focuses on the use of potential osmolyte precursor compounds for osmotic stress alleviation in granular anaerobic sludges upon exposure to hypersalinity shocks. Glutamic acid, aspartic acid, lysine, potassium, gelatine, and tryptone were tested for their potential to alleviate osmotic stress in laboratory grown and full - scale granular sludge. The laboratory grown granular sludge was adapted to 5 (R5) and 20 (R20) g Na+/L. Full-scale granular sludge was obtained from internal circulation reactors treating tannery (waste)water with influent conductivity of 29.2 (Do) and 14.1 (Li) mS/cm. In batch experiments which focused on specific methanogenic activity (SMA), R5 granular sludge was exposed to a hypersalinity shock of 20 g Na+/L. The granular sludge of Do and Li was exposed to a hypersalinity shock of 10 g Na+/L with sodium acetate as the sole carbon source. The effects on R20 granular sludge were studied at the salinity level to which the sludge was already adapted, namely 20 g Na+/L. Dosing of glutamic acid, aspartic acid, gelatine, and tryptone resulted in increased SMA compared to only acetate fed batches. In batches with added glutamic acid, the SMA increased by 115% (Li), 35% (Do) and 9% (R20). With added aspartic acid, SMA increased by 72% (Li), 26% (Do), 12% (R5) and 7% (R20). The addition of tryptone resulted in SMA increases of 36% (R5), 17% (R20), 179% (Li), and 48% (Do), whereas added gelatine increased the SMA by 30% (R5), 14% (R20), 23% (Li), and 13% (Do). The addition of lysine, meanwhile, gave negative effects on SMA of all tested granular sludges. Potassium at sea water Na/K ratio (27.8 w/w) had a slight positive effect on SMA of Do (7.3%) and Li (10.1%), whereas at double the sea water ratio (13.9% w/w) had no pronounced positive effect. R20 granular sludge was also exposed to hyposalinity shock from 20 down to 5 g Na+/L. Glutamate and N-acetyl-ß-lysine were excreted by microbial consortium in anaerobic granular sludge adapted to 20 g Na+/L upon this exposure to hyposalinity. A potential consequence when applying these results is that saline streams containing specific and hydrolysable proteins can be anaerobically treated without additional dosing of osmolytes.

Fast anaerobic sludge granulation at elevated salinity.

It is commonly accepted that high salt concentrations negatively affect microbial activity in biological wastewater treatment reactors such as upflow anaerobic sludge blanket (UASB) reactors. Microbial aggregation in such reactors is equally important. It is well documented that anaerobic granules, when exposed to high salinity become weak and disintegrate, causing wash-out, operational problems and decreasing process performance. In this research, the possibility of microbial granule formation from dispersed biomass was investigated at salinity levels of 5 and 20 g Na+/L. High removal efficiencies of soluble influent organics were achieved at both salinity levels and this was accompanied by fast and robust formation of microbial granules. The process was found to be stable for the entire operational period of 217 days. As far as we know this is the first time it has been demonstrated that stable granule formation is possible at a salinity level as high as 20 g Na+/L. Methanosaeta was identified as the dominant methanogen at both salinity levels. Streptococcus spp. and bacteria belonging to the family Lachnospiraceae were identified as the dominant microbial population at 5 and 20 and g Na+/L, respectively.