scholarly journals Development of a Rapid Assay for Determining the Relative Abundance of Bacteria

2005 ◽  
Vol 71 (12) ◽  
pp. 8481-8490 ◽  
Author(s):  
Arlene K. Rowan ◽  
Russell J. Davenport ◽  
Jason R. Snape ◽  
David Fearnside ◽  
Michael R. Barer ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Microbial Populations ◽  
Ammonia Oxidizing Bacteria ◽  
Bacterial Cells ◽  
Target Region ◽  
E Coli ◽  
Sandwich Hybridization ◽  
Sandwich Hybridization Assay ◽  
Rrna Sequences

ABSTRACT A sandwich hybridization assay for high-throughput, rapid, simple, and inexpensive quantification of specific microbial populations was evaluated. The assay is based on the hybridization of a target rRNA with differentially labeled capture and detector probes. Betaproteobacterial ammonia-oxidizing bacteria (AOB) were selected as the target group for the study, since they represent a phylogenetically coherent group of organisms that perform a well-defined geochemical function in natural and engineered environments. Reagent concentrations, probe combinations, and washing, blocking, and hybridization conditions were optimized to improve signal and reduce background. The detection limits for the optimized RNA assay were equivalent to approximately 103 to 104 and 104 to 105 bacterial cells, respectively, for E. coli rRNA and RNA extracted from activated sludge, by using probes targeting the majority of bacteria. Furthermore, the RNA assay had good specificity, permitted discrimination of rRNA sequences that differed by a 2-bp mismatch in the probe target region, and could distinguish the sizes of AOB populations in nitrifying and nonnitrifying wastewater treatment plants.

scholarly journals Ultrafiltration Process in Disinfection and Advanced Treatment of Tertiary Treated Wastewater

Membranes ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 221
Author(s):  
Rafał Tytus Bray ◽  
Katarzyna Jankowska ◽  
Eliza Kulbat ◽  
Aneta Łuczkiewicz ◽  
Aleksandra Sokołowska
Keyword(s):  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Microscopic Analysis ◽  
Fecal Coliforms ◽  
Treated Wastewater ◽  
Chemical Parameters ◽  
Virus Particles ◽  
E Coli ◽  
The One ◽  
Physical And Chemical

The paper presents the results of research on the use of ultrafiltration, using membranes of 200 and 400 kDa separation, for disinfection of municipal treated wastewater. The research was conducted on a fractional technical scale using real municipal treated wastewater from two large wastewater treatment plants treating most of the wastewater over the one-million polycentric Gdańsk agglomeration (1.2 million inhabitants). UF 200 kDa and UF 400 kDa processes enabled further improvement of the physical and chemical parameters of treated wastewater. Total phosphorus (to below 0.2 mg/L–UF 200 kDa, 0.13 mg/L–UF 400 kDa) and turbid substances (to below 0.2 mg/L, both membranes) were removed in the highest degree. COD was reduced efficiently (to below 25.6 mgO2/L–UF 200 kDa, 26.8 mgO2/L–UF 400 kDa), while total nitrogen was removed to a small extent (to 7.12 mg/L–UF 200 kDa and 5.7 mg/L–UF 400 kDa. Based on the reduction of indicator bacteria; fecal coliforms including E. coli (FC) and fecal enterococci (FE) it was found that the ultrafiltration is an effective method of disinfection. Not much indicator bacterial were observed in the permeate after processes (UF 200 kDa; FC—5 CFU/L; FE—1 CFU/L and UF 400 kDa; FC—70 CFU/L; FE—10 CFU/L. However, microscopic analysis of prokaryotic cells and virus particles showed their presence after the application of both membrane types; TCN 3.0 × 102 cells/mL–UF 200 kDa, 5.0 × 103 cells/mL–UF 400 kDa, VP 1.0 × 105/mL. The presence of potentially pathogenic, highly infectious virus particles means that ultrafiltration cannot be considered a sufficient disinfection method for treated wastewater diverted for reuse or discharged from high load wastewater treatment plants to recreational areas. For full microbiological safety it would be advisable to apply an additional disinfection method (e.g., ozonation).

Removal improvement of bacteria (Escherichia coli and enterococci) in maturation ponds using baffles

2012 ◽  
Vol 65 (4) ◽  
pp. 589-595 ◽  
Author(s):  
A. Ouali ◽  
H. Jupsin ◽  
J. L. Vasel ◽  
L. Marouani ◽  
A. Ghrabi
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Treatment Plant ◽  
E Coli ◽  
Conventional Activated Sludge ◽  
In Series ◽  
Hydrodynamic Study ◽  
Rhodamine Wt ◽  
Engineering Intervention

Korba wastewater treatment plant is a conventional activated sludge followed by three maturation ponds (MP1, MP2, MP3) in series acting as a tertiary treatment. The first study of wastewater treatment plants showed that the effluent concentration of Escherichia coli and enterococci at the outlet of the (MP3) varies between 103 and 104CFU/100 ml. After the hydrodynamic study conducted by Rhodamine WT which showed short-circuiting in the MP1, two baffles were introduced in the first maturation pond (MP1) to improve the hydrodynamic and the sanitary performances. The second hydraulic study showed that the dispersion number ‘d’ was reduced from 1.45 to 0.43 by this engineering intervention and the Peclet number was raised from 0.69 to 2.32. The hydraulic retention time was increased by 14 h. Because of well-designed baffles, the removal efficiency of E. coli and enterococci was raised between 0.2 and 0.7 log units for the first maturation pond.

scholarly journals ESKAPE Bacteria and Extended-Spectrum-β-Lactamase-Producing Escherichia coli Isolated from Wastewater and Process Water from German Poultry Slaughterhouses

2020 ◽  
Vol 86 (8) ◽  
Author(s):  
Mykhailo Savin ◽  
Gabriele Bierbaum ◽  
Jens Andre Hammerl ◽  
Céline Heinemann ◽  
Marijo Parcina ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Resistance Genes ◽  
Wastewater Treatment Plants ◽  
Process Water ◽  
Resistant Bacteria ◽  
Content Type ◽  
E Coli ◽  
Extended Spectrum ◽  
Clinically Relevant Bacteria

ABSTRACT The wastewater of livestock slaughterhouses is considered a source of antimicrobial-resistant bacteria with clinical relevance and may thus be important for their dissemination into the environment. To get an overview of their occurrence and characteristics, we investigated process water (n = 50) from delivery and unclean areas as well as wastewater (n = 32) from the in-house wastewater treatment plants (WWTPs) of two German poultry slaughterhouses (slaughterhouses S1 and S2). The samples were screened for ESKAPE bacteria (Enterococcus spp., Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) and Escherichia coli. Their antimicrobial resistance phenotypes and the presence of extended-spectrum-β-lactamase (ESBL), carbapenemase, and mobilizable colistin resistance genes were determined. Selected ESKAPE bacteria were epidemiologically classified using different molecular typing techniques. At least one of the target species was detected in 87.5% (n = 28/32) of the wastewater samples and 86.0% (n = 43/50) of the process water samples. The vast majority of the recovered isolates (94.9%, n = 448/472) was represented by E. coli (39.4%), the A. calcoaceticus-A. baumannii (ACB) complex (32.4%), S. aureus (12.3%), and K. pneumoniae (10.8%), which were widely distributed in the delivery and unclean areas of the individual slaughterhouses, including their wastewater effluents. Enterobacter spp., Enterococcus spp., and P. aeruginosa were less abundant and made up 5.1% of the isolates. Phenotypic and genotypic analyses revealed that the recovered isolates exhibited diverse resistance phenotypes and β-lactamase genes. In conclusion, wastewater effluents from the investigated poultry slaughterhouses exhibited clinically relevant bacteria (E. coli, methicillin-resistant S. aureus, K. pneumoniae, and species of the ACB and Enterobacter cloacae complexes) that contribute to the dissemination of clinically relevant resistances (i.e., blaCTX-M or blaSHV and mcr-1) in the environment. IMPORTANCE Bacteria from livestock may be opportunistic pathogens and carriers of clinically relevant resistance genes, as many antimicrobials are used in both veterinary and human medicine. They may be released into the environment from wastewater treatment plants (WWTPs), which are influenced by wastewater from slaughterhouses, thereby endangering public health. Moreover, process water that accumulates during the slaughtering of poultry is an important reservoir for livestock-associated multidrug-resistant bacteria and may serve as a vector of transmission to occupationally exposed slaughterhouse employees. Mitigation solutions aimed at the reduction of the bacterial discharge into the production water circuit as well as interventions against their further transmission and dissemination need to be elaborated. Furthermore, the efficacy of in-house WWTPs needs to be questioned. Reliable data on the occurrence and diversity of clinically relevant bacteria within the slaughtering production chain and in the WWTP effluents in Germany will help to assess their impact on public and environmental health.

Novel Nitrospira-like bacteria as dominant nitrite-oxidizers in biofilms from wastewater treatment plants: diversity and in situ physiology

2000 ◽  
Vol 41 (4-5) ◽  
pp. 85-90 ◽  
Author(s):  
H. Daims ◽  
P.H. Nielsen ◽  
J.L. Nielsen ◽  
S. Juretschko ◽  
M. Wagner
Keyword(s):  
Wastewater Treatment ◽  
In Situ Hybridization ◽  
16S Rrna ◽  
Fluorescent In Situ Hybridization ◽  
Wastewater Treatment Plants ◽  
Combined Application ◽  
Fundamental Understanding ◽  
Unculturable Bacteria ◽  
Rrna Sequences

The frequency and distribution of putatively nitrite-oxidizing, Nitrospira- like bacteria in nitrifying biofilms from two reactors receiving wastewater with different ammonia and salt concentrations were observed by fluorescent in situ hybridization. For this purpose, new 16S rRNA-directed oligonucleotide probes targeting the bacterial phylum Nitrospira and the three main lineages within this phylum were developed and evaluated. The diversity of Nitrospira-like bacteria in the reactors was additionally investigated by retrieval and comparative analysis of full 16S rRNA sequences from the biofilms. We found that, despite of the differences in the influent composition, Nitrospira-like bacteria form dominant populations in both reactors. In addition, first insights into the physiology of these still unculturable bacteria were obtained by the incubation of active biofilm samples with radioactively labeled substrates followed by the combined application of fluorescent in situ hybridization and microautoradiography. The results are discussed in consideration of the frequently observed dominance of Nitrospira-like bacteria in nitrifying bioreactors. Consequently, high priority should be assigned to future studies on the ecology and physiology of these organisms in order to increase our fundamental understanding of nitrogen cycling and to enable knowledge-driven future improvements of nitrifying wastewater treatment plants.

scholarly journals Removal of antibiotic resistant E. coli in two Norwegian wastewater treatment plants and by nano- and ultra-filtration processes

2017 ◽  
Vol 77 (4) ◽  
pp. 1115-1126 ◽  
Author(s):  
Carsten Ulrich Schwermer ◽  
Pawel Krzeminski ◽  
Aina Charlotte Wennberg ◽  
Christian Vogelsang ◽  
Wolfgang Uhl
Keyword(s):  
Wastewater Treatment ◽  
Membrane Filtration ◽  
Municipal Wastewater ◽  
Wastewater Treatment Plants ◽  
Considerable Change ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Physical Chemical ◽  
Ultra Filtration

Abstract The effectivity of different treatment stages at two large wastewater treatment plants (WWTPs) located in Oslo, Norway, to remove antibiotic resistant Escherichia coli from municipal wastewater was investigated. The WWTPs were effective in reducing the total cultivable E. coli. The E. coli in WWTP samples were mainly resistant to ampicillin (6–27%) and trimethoprim-sulfamethoxazole (5–24%), and, to a lesser extent, tetracycline (3–14%) and ciprofloxacin (0–7%). In the first WWTP, a clear decrease in the percentage of E. coli resistant to these antibiotics was found, with the main removal occurring during physical/chemical treatment. In the second WWTP, the percentage of cultivable resistant E. coli did not display a considerable change. During laboratory-scale membrane filtration of WWTP effluents using ultrafiltration (UF) and nanofiltration (NF) membranes, all E. coli, including those resistant to antibiotics, were removed completely. The results imply that UF and NF processes are potent measures to remove antibiotic resistant bacteria (ARB) during post-treatment of WWTP effluents, thus reducing the potential spread of antibiotic resistance in the receiving aquatic environment.

Enumeration of viable Escherichia coli by real-time PCR with propidium monoazide

2012 ◽  
Vol 66 (10) ◽  
pp. 2065-2073 ◽  
Author(s):  
N. Yokomachi ◽  
J. Yaguchi
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Real Time ◽  
Real Time Pcr ◽  
Wastewater Treatment Plants ◽  
Propidium Monoazide ◽  
E Coli ◽  
Cell Counts ◽  
Viable Cells ◽  
Heat Treated

A photo-inducible DNA-binding dye, propidium monoazide (PMA), was used to distinguish viable and dead Escherichia coli cells. Microscopic observations using a combination of the dyes 4′,6-diamidino-2-phenylindole and PMA indicated that PMA stained only dead cells, with membrane damage, red. Mixtures of viable and heat-treated E. coli cells were subjected to real-time polymerase chain reaction (PCR) with PMA treatment. Viable cell counts were linearly related to real-time PCR threshold cycle values for PMA-treated cells in the mixtures of viable and heat-treated cells, as long as the ratio of dead cells to viable cells was no greater than 10. In the wastewater treatment plants, total, viable and culturable E. coli were enumerated by real-time PCR, real-time PCR coupled with PMA treatment and the most probable number method using EC-MUG medium, respectively. The concentrations of viable E. coli in the wastewater treatment plants were much higher than those of culturable cells. In addition, viable cells were even more chlorine resistant than culturable ones.

Long-term effects of engineered nanoparticles on enzyme activity and functional bacteria in wastewater treatment plants

2015 ◽  
Vol 72 (1) ◽  
pp. 99-105 ◽  
Author(s):  
Xiong Zheng ◽  
Haining Huang ◽  
Yinglong Su ◽  
Yuanyuan Wei ◽  
Yinguang Chen
Keyword(s):  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Engineered Nanoparticles ◽  
Ammonia Oxidizing Bacteria ◽  
Long Term Effects ◽  
Gene Expressions ◽  
Key Enzymes ◽  
Functional Bacteria ◽  
Wide Range

Abstract The pervasive use of engineered nanoparticles (NPs) in a wide range of fields raises concerns about their potential environmental impacts. Previous studies confirmed that some NPs had already entered wastewater treatment plants (WWTPs). Wastewater nutrient removal depends on the metabolisms of activated sludge bacteria and their related key enzymes. Therefore, this study compared the possible influences of Al2O3, SiO2, TiO2, and ZnO NPs on the key enzymes activities and microbial community structures involved in wastewater treatment facilities. It was found that long-term exposure to these NPs significantly affected the microbial communities and changed the relative abundances of key functional bacteria, such as ammonia-oxidizing bacteria. Also, the gene expressions and catalytic activities of essential enzymes, such as ammonia monooxygenase, nitrite oxidoreductase, nitrate reductase, and nitrite reductase, were decreased, which finally resulted in a lower efficiency of biological nitrogen removal.

scholarly journals Simultaneous Fluorescent Gram Staining and Activity Assessment of Activated Sludge Bacteria

2002 ◽  
Vol 68 (10) ◽  
pp. 4772-4779 ◽  
Author(s):  
Scott Forster ◽  
Jason R. Snape ◽  
Hilary M. Lappin-Scott ◽  
Jonathan Porter
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Flow Cytometric Analysis ◽  
Quantitative Measure ◽  
Bacterial Cells ◽  
Gram Positive ◽  
Gram Negative ◽  
Highly Active ◽  
Gram Staining

ABSTRACT Wastewater treatment is one of the most important commercial biotechnological processes, and yet the component bacterial populations and their associated metabolic activities are poorly understood. The novel fluorescent dye hexidium iodide allows assessment of Gram status by differential absorption through bacterial cell walls. Differentiation between gram-positive and gram-negative wastewater bacteria was achieved after flow cytometric analysis. This study shows that the relative proportions of gram-positive and gram-negative bacterial cells identified by traditional microscopy and hexidium iodide staining were not significantly different. Dual staining of cells for Gram status and activity proved effective in analyzing mixtures of cultured bacteria and wastewater populations. Levels of highly active organisms at two wastewater treatment plants, both gram positive and gram negative, ranged from 1.5% in activated sludge flocs to 16% in the activated sludge fluid. Gram-positive organisms comprised <5% of the total bacterial numbers but accounted for 19 and 55% of the highly active organisms within flocs at the two plants. Assessment of Gram status and activity within activated sludge samples over a 4-day period showed significant differences over time. This method provides a rapid, quantitative measure of Gram status linked with in situ activity within wastewater systems.

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