Biofilm microbial community structure in an urban lake utilizing reclaimed water

Analyses of biofilm community structure may potentially be employed for aquatic ecosystem health assessment, however, to date, biofilm diversity within urban lakes using reclaimed water has not been examined. Accordingly, the microbial community diversity and structure of biofilms from the surface of multiple matrices with varying roughness (0.1, 1.0 and 10.0 μm) were characterized using a suite of molecular techniques including scanning electron microscopy, genetic fingerprinting and phospholipid-derived fatty acid analyses. Samples were largely comprised of inorganic particles, algae and numerous bacterial species; 12 phospholipid-derived fatty acid (PLFA) types were identified, significantly less than typically associated with sewage. Both growth matrix surface roughness and biofilm growth phase were shown to concur with significantly different microbial quantity and community structures. Gram-negative bacteria bacillus i15:03OH and 18:0 were the dominant bacterial genera, collectively comprising ≈75 % of identified PLFA species content. Calculated species diversity (H) and species dominance (D) exhibited identical correlational patterns with measured water quality parameters; significant positive correlations were exhibited with respect to Mg2, while significant negative correlations were found for NO3, TP, BOD, COD, SP, PO4, SO4 and pH. Results indicate that analyses of biofilm formation and structure could be effectively used to undertake integrated assessments of the ecological health of lake systems using reclaimed water. Further work is required to elucidate the optimum conditions for sample collection and analytical interpretation

Reconstruction of auto-tissue-engineered lamellar cornea by dynamic culture for transplantation: a rabbit model.

To construct an auto-tissue-engineered lamellar cornea (ATELC) for transplantation, based on acellular porcine corneal stroma and autologous corneal limbal explants, a dynamic culture process, which composed of a submersion culture, a perfusion culture and a dynamic air-liquid interface culture, was performed using appropriate parameters. The results showed that the ATELC-Dynamic possessed histological structure and DNA content that were similar to native lamellar cornea (NLC, p>0.05). Compared to NLC, the protein contents of zonula occludens-1, desmocollin-2 and integrin β4 in ATELC-Dynamic reached 93%, 89% and 73%, respectively. The basal cells of ATELC-Dynamic showed a better differentiation phenotype (K3-, P63+, ABCG2+) compared with that of ATELC in static air-lift culture (ATELC-Static, K3+, P63-, ABCG2-). Accordingly, the cell-cloning efficiency of ATELC-Dynamic (9.72±3.5%) was significantly higher than that of ATELC-Static (2.13±1.46%, p<0.05). The levels of trans-epithelial electrical resistance, light transmittance and areal modulus variation in ATELC-Dynamic all reached those of NLC (p>0.05). Rabbit lamellar keratoplasty showed that the barrier function of ATELC-Dynamic was intact, and there were no signs of epithelial shedding or neovascularization. Furthermore, the ATELC-Dynamic group had similar optical properties and wound healing processes compared with the NLC group. Thus, the sequential dynamic culture process that was designed according to corneal physiological characteristics could successfully reconstruct an auto-lamellar cornea with favorable morphological characteristics and satisfactory physiological function

Physiological function of ATELC-Dynamic in a rabbit lamellar keratoplasty model.

<p>(A) Postoperative observation of lamellar keratoplasty. (B) The light transmittance of transplanted lamellar cornea over the wavelength range of 300–800 nm at 20 days. (C) DiO-labeled seeding cells at 7 days, and the expression of collagen III at 20 days.</p

The major components of the basement membrane in APCS (↓: basement membrane components, green: collagen IV, laminin, fibronectin and collagen VII, red: collagen I).

<p>The major components of the basement membrane in APCS (↓: basement membrane components, green: collagen IV, laminin, fibronectin and collagen VII, red: collagen I).</p

The construction process of auto-tissue-engineered lamellar cornea.

<p>(A) Submersion culture (5 days). (B) Perfusion culture (3 days). (C) Dynamic air-liquid interface culture (with perfusion culture on the bottom surface, 3 days). (D) Dynamic air-liquid interface culture (with a dry chamber on the bottom surface, 12 hours). (E) Macroscopic view of ATELC-Dynamic in each stage of the construction process.</p

The changes in histomorphology and DNA content during the construction process (▴: connection between neighbor cells, *: cellular nucleus,↓: desmosome).

<p>(A) ATELC-Dynamic, after submersion culture. (B) ATELC-Dynamic, after perfusion culture. (C) ATELC-Dynamic, after dynamic air-liquid interface culture. (D) Negative control: ATELC-Static. (E) Positive control: NLC. (F) DNA contents of the NLC, ATELC-Dynamic, and ATELC-Static groups.</p

Biofilm microbial community structure in an urban lake utilizing reclaimed water

Analyses of biofilm community structure may potentially be employed for aquatic ecosystem health assessment, however, to date, biofilm diversity within urban lakes using reclaimed water has not been examined. Accordingly, the microbial community diversity and structure of biofilms from the surface of multiple matrices with varying roughness (0.1, 1.0 and 10.0 μm) were characterized using a suite of molecular techniques including scanning electron microscopy, genetic fingerprinting and phospholipid-derived fatty acid analyses. Samples were largely comprised of inorganic particles, algae and numerous bacterial species; 12 phospholipid-derived fatty acid (PLFA) types were identified, significantly less than typically associated with sewage. Both growth matrix surface roughness and biofilm growth phase were shown to concur with significantly different microbial quantity and community structures. Gram-negative bacteria bacillus i15:03OH and 18:0 were the dominant bacterial genera, collectively comprising ≈75 % of identified PLFA species content. Calculated species diversity (H) and species dominance (D) exhibited identical correlational patterns with measured water quality parameters; significant positive correlations were exhibited with respect to Mg2, while significant negative correlations were found for NO3, TP, BOD, COD, SP, PO4, SO4 and pH. Results indicate that analyses of biofilm formation and structure could be effectively used to undertake integrated assessments of the ecological health of lake systems using reclaimed water. Further work is required to elucidate the optimum conditions for sample collection and analytical interpretation