Plant-derived pectin nanocoatings to prevent inflammatory cellular response of osteoblasts following Porphyromonas gingivalis infection

Background: Bioengineered plant-derived Rhamnogalacturonan-Is (RG-Is) from pectins are potential candidates for surface nanocoating of medical devices. It has recently been reported that RG-I nanocoatings may prevent bacterial infection and improve the biocompatibility of implants. The aim of the study was to evaluate in vitro impact of bioengineered RG-I nanocoatings on osteogenic capacity and proinflammatory cytokine response of murine osteoblasts following Porphyromonas gingivalis infection. Methods: Murine MC3T3-E1 osteoblasts and isolated primary calvarial osteoblasts from C57BL/6J (B6J osteoblasts) mice were infected with P. gingivalis and incubated on tissue culture polystyrene plates with or without nanocoatings of unmodified RG-Is isolated from potato pulps (PU) or dearabinanated RG-Is (PA). To investigate a behavior of infected osteoblasts cultured on RG-Is cell morphology, proliferation, metabolic activity, mineralization and osteogenic and pro-inflammatory gene expression were examined. Results: Following P. gingivalis infection, PA, but not PU, significantly promoted MC3T3-E1 and BJ6 osteoblasts proliferation, metabolic activity, and calcium deposition. Moreover, Il-1b, Il-6, TNF-α, and Rankl gene expressions were downregulated in cells cultured on PU and to a higher extent on PA as compared to the corresponding control, whereas Runx, Alpl, Col1a1, and Bglap gene expressions were upregulated vice versa. Conclusion: Our data clearly showed that pectin RG-Is nanocoating with high content of galactan (PA) reduces the osteoblastic response to P. gingivalis infection in vitro and may, therefore, reduce a risk of inflammation especially in immunocompromised patients with rheumatoid or periodontal disorders

Wpływ polimerów zewnątrzkomórkowych na granulację osadu czynnego w warunkach tlenowych

Badano wpływ polimerów zewnątrzkomórkowych na proces biogranulacji. Po 7 dnia trwania eksperymentu obserwowano pojawianie się tlenowych biogranul osadu czynnego, Zaobserwowano, że wraz ze wzrostem produkcji substancji EPS następował wzrost średnic granul. Dominującym składnikiem substancji EPS w granulach były białka. W porównaniu do luźno związanych substancji EPS (LB-EPS), mocno związane EPS (TB-EPS) miały znaczący udział w formowaniu się granul. Wykazano, że polisacharydy stanowią ważny składnik strukturalny granul

Influence of calcium, magnesium and iron ions on the molecular mass of exoproteins during biogranulation

In this study, we performed the qualitative analysis of exoproteins during granule formation in the presence or in the absence of cations. The staining of thin granule cryosections showed that nucleic acids, proteins, polysaccharides and calcium cations were the dominant components of the granules. Proteins are the structural components associated with calcium ions. We determined changes in the proteomic profile and tightly bound extracellular polymeric substances (EPS) of the slime. The exopolymeric matrix containing the proteins was extracted using the Dowex resin method. Proteomic profile was analysed by SDS-PAGE method (sodium dodecyl sulphate polyacrylamide gel electrophoresis) using Coomassie blue staining in the samples of the aerobic granule matrix formed in the presence of multivalent cations and compared with that of the aerobic granules cultivated without cations. The results indicate that the granule matrix is predominantly composed of large and complex proteins that are tightly bound within the granular structure. The tightly bound extracellular polymeric substances (TB-EPS) may play a role in improved mechanical stability of aerobic granules. In the supernatant fraction of the sludge, only a small amount of free proteins in the medium molecular mass range was detected. The protein with high molecular mass ( 116 kDa) produced in the reactors with added Ca2+. Ca2+ had a considerable regulatory influence on production of extracellular proteins during aerobic granulation

Detection and characteristics of sulfamethoxazole-resistant bacteria in constructed wetlands treating sulfamethoxazole-rich wastewater

Constructed wetlands (CW) are one of the biological wastewater treatment systems that reflect the natural processes occurring in swamps. Constructed wetlands use microbiological and physico-chemical processes as well as plant metabolism in order to purify wastewater. In such treatment systems, the role of microorganisms is crucial. In this experiment, synthetic communal wastewater containing sulfamethoxazole (SMX; chemotherapeutic) at a concentration of 5 mg/l was applied in the CW systems, both unplanted and planted with Phalaris arundinacea , also known as reed canary grass. Fourteen Gram-positive SMX-resistant bacteria strains were isolated from the CW column fillings and the plant rhizosphere. All of these were identified as representatives of Bacillus sp . based on 16S rRNA sequencing. Despite this molecular identification, the isolates differed significantly in their biochemical features. All 14 isolates presented resistance toward sulfamethoxazole and all 14 strains possessed a sul1 gene, while only 4 gave positive results in sul2 and 3 in sul3 PCR tests. None of the isolated strains possessed all three sul genes. A PCR-DGGE based analysis of the presence of SMX-resistant bacteria in the CW community was undertaken. It was found that none of the isolates represented the dominant genotype in the bacterial community

Detection and characteristics of sulfamethoxazole-resistant bacteria in constructed wetlands treating sulfamethoxazole-rich wastewater

Constructed wetlands (CW) are one of the biological wastewater treatment systems that reflect the natural processes occurring in swamps. Constructed wetlands use microbiological and physico-chemical processes as well as plant metabolism in order to purify wastewater. In such treatment systems, the role of microorganisms is crucial. In this experiment, synthetic communal wastewater containing sulfamethoxazole (SMX; chemotherapeutic) at a concentration of 5 mg/l was applied in the CW systems, both unplanted and planted with Phalaris arundinacea , also known as reed canary grass. Fourteen Gram-positive SMX-resistant bacteria strains were isolated from the CW column fillings and the plant rhizosphere. All of these were identified as representatives of Bacillus sp . based on 16S rRNA sequencing. Despite this molecular identification, the isolates differed significantly in their biochemical features. All 14 isolates presented resistance toward sulfamethoxazole and all 14 strains possessed a sul1 gene, while only 4 gave positive results in sul2 and 3 in sul3 PCR tests. None of the isolated strains possessed all three sul genes. A PCR-DGGE based analysis of the presence of SMX-resistant bacteria in the CW community was undertaken. It was found that none of the isolates represented the dominant genotype in the bacterial community

Qualitative variability in microbial community of constructed wetlands used for purifying wastewater contaminated with pharmaceutical substances

Pharmaceutical substances and their residues are increasingly present in the environment. Therefore, attempts at their removal are made by using different processes. Increasingly important among these processes are those modeled on natural phenomena which occur in wetland ecosystems, called technical scale constructed wetlands. Microbial degradation is an important process in these constructed wetlands. The biodegradation of chemicals often involves a complex series of biochemical reactions and usually varies with the microorganisms involved. The objectives of this study were to determine the impact of sulfamethoxazole and diclofenac on ammonia oxidizing bacteria and other parameters of wastewater in the microcosm of down-flow constructed wetlands. The Spearman correlation coefficient attained negative values in the case of comparison of the Shannon biodiversity index and the parameters of purified wastewater. This dependence was pronounced. In the case of pharmaceutical substances dosed with wastewater, the Spearman correlation coefficient assumed positive values. The highest value assumed by the Spearman correlation coefficient (0.9) was for the removal of diclofenac and Shannon index values for the planted columns, with a very high relationship. For unplanted columns, this value equaled 0.6. For sulfamethoxazole, the value for planted columns was 0.7, and for unplanted -0.7. The presence of plants did not have an impact on the Shannon biodiversity index

Enrichment of endophytic Actinobacteria in roots and rhizomes of <i>Miscanthus × giganteus</i> plants exposed to diclofenac and sulfamethoxazole

This study investigates how wastewater containing 2 mg l(-1) of sulfamethoxazole (SMX) and 2 mg l(-1) of diclofenac (DCF) affects the composition of bacterial communities present in the roots and rhizomes of Miscanthus x giganteus plants grown in laboratory-scale constructed wetlands. Bacterial communities in plant roots and rhizomes were identified in treated and control samples by 16S rRNA amplicon sequencing. Moreover, bacterial endophytes were isolated in R2A and 1/10 869 media and screened for their ability to metabolize SMX and DCF in liquid medium by HPLC. Our results show significant changes in the abundance of main genera, namely Sphingobium and Streptomyces between control and treated plants. Around 70% of the strains isolated from exposed plants belonged to the phylum Actinobacteria and were classified as Streptomyces, Microbacterium, and Glycomyces. In non-exposed plants, Proteobacteria represented 43.5% to 63.6% of the total. We identified 17 strains able to remove SMX and DCF in vitro. From those, 76% were isolated from exposed plants. Classified mainly as Streptomyces, they showed the highest SMX (33%) and DCF (41%) removal efficiency. These isolates, alone or in combination, might be used as bio-inoculants in constructed wetlands to enhance the phytoremediation of SMX and DCF during wastewater treatment

The effect of temperature on the efficiency of industrial wastewater nitrification and its (geno)toxicity

The paper deals with the problem of the determination of the effects of temperature on the efficiency of the nitrification process of industrial wastewater, as well as its toxicity to the test organisms. The study on nitrification efficiency was performed using wastewater from one of Polish chemical factories. The chemical factory produces nitrogen fertilizers and various chemicals. The investigated wastewater was taken from the influent to the industrial mechanical-biological wastewater treatment plant (WWTP). The WWTP guaranteed high removal efficiency of organic compounds defined as chemical oxygen demand (COD) but periodical failure of nitrification performance was noted in last years of the WWTP operation. The research aim was to establish the cause of recurring failures of nitrification process in the above mentioned WWTP. The tested wastewater was not acutely toxic to activated sludge microorganisms. However, the wastewater was genotoxic to activated sludge microorganisms and the genotoxicity was greater in winter than in spring time. Analysis of almost 3 years’ period of the WWTP operation data and laboratory batch tests showed that activated sludge from the WWTP under study is very sensitive to temperature changes and the nitrification efficiency collapses rapidly under 16°C. Additionally, it was calculated that in order to provide the stable nitrification, in winter period the sludge age (SRT) in the WWTP should be higher than 35 days

Removal of diclofenac and sulfamethoxazole from synthetic municipal waste water in microcosm downflow constructed wetlands: Start-up results

<div><p>ABSTRACT</p><p>The objectives of this study were to investigate the start-up removal of pharmaceutical compounds diclofenac and sulfamethoxazole in microcosm downflow constructed wetlands and their effect on the performance of the studied constructed wetlands, and also to assess the effect of plants on the removal of these compounds. The experimental system that was used in this 86-day experiment consisted of 24 columns filled up to 70 cm with predominantly sandy material. Four types of columns were used (six replicates) depending on the presence of plants (<i>Phalaris arundinacea</i> L. var. <i>picta</i> L.) and the presence of pharmaceutical compounds in the influent. The influent was synthetic municipal waste water to which a mixture of 5 mg/L of diclofenac and 5 mg/L of sulfamethoxazole was added. The observed removal of diclofenac was moderate (approx. 50%) and the removal of sulfamethoxazole was relatively low (24–30%). It was found that the removal of diclofenac and sulfamethoxazole was not affected by the vegetation. The presence of diclofenac and sulfamethoxazole in the influent had significant effect on the effluent concentration of N-NO₃ and the water loss in the columns, which in both cases were lower than in the control columns. The scope for further research was discussed.</p></div