Mycobacteria in drinking water distribution systems: ecology and significance for human health

In contrast to the notorious pathogens Mycobacterium tuberculosis and M. leprae, the majority of the mycobacterial species described to date are generally not considered as obligate human pathogens. The natural reservoirs of these non-primary pathogenic mycobacteria include aquatic and terrestrial environments. Under certain circumstances, e.g., skin lesions, pulmonary or immune dysfunctions and chronic diseases, these environmental mycobacteria (EM) may cause disease. EM such as M. avium, M. kansasii, and M. xenopi have frequently been isolated from drinking water and hospital water distribution systems. Biofilm formation, amoeba-associated lifestyle, and resistance to chlorine have been recognized as important factors that contribute to the survival, colonization and persistence of EM in water distribution systems. Although the presence of EM in tap water has been linked to nosocomial infections and pseudo-infections, it remains unclear if these EM provide a health risk for immunocompromised people, in particular AIDS patients. In this regard, control strategies based on maintenance of an effective disinfectant residual and low concentration of nutrients have been proposed to keep EM numbers to a minimum in water distribution systems.     

Influence of phosphorus on biofilm formation in model drinking water distribution systems

Aims:  This study investigated the effects of phosphorus on biofilm formation via annular reactor systems in terms of biofilm cell growth, exopolysaccharide (EPS) production, biofilm structure and cell metabolic potential.
Methods and Results:  Drinking water biofilms were developed in annular reactors with supplement of carbon and different levels of phosphorus. The biofilm formation was monitored over a period of 30 days. Biofilm related parameters were examined by various methods, which included heterotrophic plate count, total carbohydrate content, confocal laser scanning microscopy and GN2 microplate assay. Our results showed that phosphorus addition can promote the biofilm cell growth (cell count increased about 1 log with addition of 30 and 300 μg l⁻¹ of phosphorus). However, the addition of 30 and 300 μg l⁻¹ of phosphorus caused 81% and 77% decrease in EPS production, respectively. The results of biofilm structure analysis showed that the addition of 30 and 300 μg l⁻¹ of phosphorus can induce thicker and less homogeneous biofilms with more biomass. Furthermore, the addition of 30 and 300 μg l⁻¹ of phosphorus dramatically increased the biofilm cell metabolic potential. The addition of 3 μg l⁻¹ of phosphorus was found to have minor effects on the parameters examined.
Conclusions:  The results indicate phosphorus addition to drinking water distribution system (DWDS) has a complicated effect on the biofilm formation.
Significance and Impact of the Study:  As the addition of phosphorus at certain levels can affect the biofilm growth in DWDS, care should be taken when phosphate-based corrosion inhibitors are used in the DWDS.     

Biofilm formation in water cooling systems

Biofilm formation on stainless steel samples immersed in cooling water has been evaluated by exposing metal samples to cooling seawater for 30 days. Anaerobic bacteria were then at 1.6 × 10⁶/cm², with sulphate-reducing species predominating. Aerobic bacteria and fungi were 2600 and 140/cm², respectively. After 60 days, numbers of aerobic microorganisms remained constant whereas the count of anaerobic microorganisms had increased to 1.8×10⁹/cm². Scanning electron microscopy showed the presence of morphologically different microorganisms in deposits and as a mucilaginous net. No signs of corrosion were detected on the stainless steel surface.The authors are with the Departamento de Engenharia Bioquimica Centro de Tecnologia, Bloco E. Universidade Federal do Rio de Janeiro Ilha do Fundão, 21941-900 Rio de Janeiro, Brazil     

Formation of biofilms in drinking water distribution networks,a case study in two cities in Finland and Latvia

The formation of biofilms in drinking water distribution networks is a significant technical, aesthetic and hygienic problem. In this study, the effects of assimilable organic carbon, microbially available phosphorus (MAP), residual chlorine, temperature and corrosion products on the formation of biofilms were studied in two full-scale water supply systems in Finland and Latvia. Biofilm collectors consisting of polyvinyl chloride pipes were installed in several waterworks and distribution networks, which were supplied with chemically precipitated surface waters and groundwater from different sources. During a 1-year study, the biofilm density was measured by heterotrophic plate counts on R2A-agar, acridine orange direct counting and ATP-analyses. A moderate level of residual chorine decreased biofilm density, whereas an increase of MAP in water and accumulated cast iron corrosion products significantly increased biofilm density. This work confirms, in a full-scale distribution system in Finland and Latvia, our earlier in vitro finding that biofilm formation is affected by the availability of phosphorus in drinking water.     

Elasticity and physico-chemical properties during drinking water biofilm formation

Atomic force microscope techniques and multi-staining fluorescence microscopy were employed to study the steps in drinking water biofilm formation. During the formation of a conditioning layer, surface hydrophobic forces increased and the range of characteristic hydrophobic forces diversified with time, becoming progressively complex in macromolecular composition, which in return triggered irreversible cellular adhesion. AFM visualization of 1 to 8 week drinking water biofilms showed a spatially discontinuous and heterogeneous distribution comprising an extensive network of filamentous fungi in which biofilm aggregates were embedded. The elastic modulus of 40-day-old biofilms ranged from 200 to 9000 kPa, and the biofilm deposits with a height >0.5 μm had an elastic modulus <600 kPa, suggesting that the drinking water biofilms were composed of a soft top layer and a basal layer with significantly higher elastic modulus values falling in the range of fungal elasticity.     

Bio-stabilization of drinking water in supply systems as observed in the tropics

Changes in the bacterial quality of drinking water were examined in the supply system of tropical Dar es Salaam, Tanzania. The water treatment fulfilled WHO standard guidelines. However, the water quality allowed for significant regrowth of bacteria, demonstrating that the WHO guidelines are insufficient to check whether or not the drinking water has an acceptable regrowth potential. An experimental pipe system was examined on site. A growth model was established, based on a zero order bacterial detachment kinetics for the biofilms colonizing the inner surface of the pipes, with a release rate factor R, and a first order growth kinetics in the water flowing in the pipes with a growth rate factor . The rate factors were estimated to be R=1.9×10⁸ m^-2 h^-1 and =2.5 h^-1. It was concluded that growth in the biofilm played a dominant role in the bio-stabilization processes, while the initial microbial quality of the water works water and the growth in the water phase were of minor importance. The bio-stabilization process is understood herein as the reduction in the concentration of nutrients and assimilable organic carbon in the water, and thus as the reduction of the growth potential of the water. Bacterial growth and recontamination were examined during storage at the user site. The results showed that heterotrophic bacteria grew, while faecal coliforms decayed during storage. It was concluded that the users handled the water hygienically.The authors are with the Technical University of Denmark, IMT, CDC-Build, 208 DK-2800 Lyngby, Denmark     

生物膜形成机理及影响因素探究

生物膜是一种依附于载体材料的特殊微生物聚集体,其大量存在于自然环境中,并在水质净化、废水处理等领域广泛应用.本文介绍了生物膜形成基本原理,综述了有关载体界面性质、胞外多聚物(EPS)关键组分对生物膜形成及其稳定性的影响,并对各学科交叉研究生物膜提供技术思路.     

城市生活饮用水中非结核分枝杆菌调查

目的 调查上海市生活饮用水中非结核分枝杆菌的状况及主要菌种分布。方法 应用过滤法收集细菌, 并在改良罗氏培养基上培养; 通过16S rRNA 测序鉴定菌种。结果 上海市生活饮用水中非结核分枝杆菌的检出率为16. 7% , 其中自来水厂原水、出厂水和居民生活饮用终端水的检出率分别为60% 、25% 和10. 3% 。分离鉴定的非结核分枝杆菌菌种为戈登分枝杆菌及偶发分枝杆菌, 分别占90% 及10% 。结论 在上海市居民生活饮用水系统中存在非结核分枝杆菌, 因此, 应当采取有效的控制方法以保护公众的健康。     

Influence of microbial interactions and EPS/polysaccharide composition on nutrient removal activity in biofilms formed by strains found in wastewater treatment systems

The study of biofilm function, structure and microbial interactions might help to improve our understanding of biofilm wastewater treatment processes. However, few reports specifically address the influence of interactions within multispecies biofilms on microbial activity and biofilm composition. Thus, the relationship between biofilm formation, denitrification activity, phosphorus removal and the composition of extracellular polymeric substances (EPS), exopolysaccharides and the bacterial community was investigated using biofilms of denitrifying and phosphorus removing strains Comamonas denitrificans 110, Brachymonas denitrificans B79, Aeromonas hydrophila L6 and Acinetobacter calcoaceticus ATCC23055. Denitrification activity within the biofilms generally increased with the amount of biofilm while phosphorus removal depended on bacterial growth rate. Synergistic effects of co-growth on denitrification (B. denitrificans B79 and A. hydrophila L6) and phosphorus removal (C. denitrificans 110 with either A. calcoaceticus or A. hydrophila L6) were observed. B. denitrificans B79 was highly affected by interspecies interactions with respect to biofilm formation, denitrification activity and EPS composition, while C. denitrificans 110 remained largely unaffected. In some of the dual and quadruple strain biofilms new exopolysaccharide monomers were detected which were not present in the pure strain samples.     

The study of fungi in drinking water

The occurrence of fungi in drinking water has received increased attention in the last decades, and fungi are now generally accepted as drinking water contaminants. The knowledge about the occurrence and diversity of fungi in water has increased considerably from a low knowledge base. However, the relevance of waterborne fungi for water quality and human health is poorly understood and still conflicting. Scientific reports on effective treatment against fungi in water are also few. This article presents a review of the literature on fungal water studies, including some general results, and considerations of significance, limits, contradictions, precautions, and practical consequences.     

Aluminium oxide nanoparticles inhibit EPS production,adhesion and biofilm formation by multidrug resistant Acinetobacter baumannii

Abstract

Acinetobacter baumannii is a biofilm forming multidrug resistant (MDR) pathogen responsible for respiratory tract infections. In this study, aluminium oxide nanoparticles (Al₂O₃ NPs) were synthesized and characterized by TEM and EDX and shown to be spherical shaped nanoparticles with a diameter < 10?nm. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) for the Al₂O₃ NPs ranged between 125 and 1,000?µg ml^?1. Exposure to NPs caused cellular membrane disruption, indicated by an increase in cellular leakage of the contents. Biofilm inhibition was 11.64 to 70.2%, whereas attachment of bacteria to polystyrene surfaces was reduced to 48.8 to 51.9% in the presence of NPs. Nanoparticles also reduced extracellular polymeric substance production and the biomass of established biofilms. The data revealed the non-toxic nature of Al₂O₃ NPs up to a concentrations of 120?µg ml^?1 in HeLa cell lines. These results demonstrate an effective and safer use of Al₂O₃ NPs against the MDR A. baumannii by targeting biofilm formation, adhesion and EPS production.     

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems

Abstract

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm² (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, “the biofilm accumulation potential”, represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.     

The effect of turbulent flow and surface roughness on biofilm formation in drinking water

There is considerable interest in both Europe and the USA in the effects of microbiological fouling on stainless steels in potable water. However, little is known about the formation and effects of biofilms, on stainless steel in potable water environments, particularly in turbulent flow regimes. Results are presented on the development of biofilms on stainless steel grades 304 and 316 after exposure to potable water at velocities of 0.32, 0.96 and 1.75 m s⁻¹. Cell counts on slides of stainless steel grades 304 and 316 with both 2B (smooth) and 2D (rough) finishes showed viable and total cell counts were higher at the higher flow rates of 0.96 and 1.75 m s⁻¹, compared to a flow rate of 0.32 m s⁻¹. Extracellular polysaccharide levels were not significantly different (P< 0.05) between each flow rate on all stainless steel surfaces studied. higher levels were found at the higher water velocities. the biofilm attached to stainless steel was comprised of a mixed bacterial flora including Acinetobacter sp, Pseudomonas spp, Methylobacterium sp, and Corynebacterium/Arthrobacter spp. Epifluorescence microscopy provided evidence of rod-shaped bacteria and the formation of stands, possibly of extracellular material attached to stainless steel at high flow rates but not at low flow rates. Received 04 February 1998/ Accepted in revised form 12 February 1999     

Potential for biofilm development in drinking water distribution systems

Regrowth of micro-organisms in drinking water distribution systems is caused by the utilisation of biodegradable compounds which are either present in treated water or originate from materials in contact with drinking water. In the Netherlands most drinking water is distributed without disinfectant residual and regrowth is limited by achieving biostable drinking water. A combination of methods is used to assess the biostability of drinking water. These methods are: (1) determination of the concentration of easily assimilable organic carbon (AOC); and (2) assessment of the biofilm formation rate (BFR). Assimilated organic carbon concentrations in drinking water in the Netherlands range from a few μg C/l in slow sand filtrates and in ground water supplies to values of ～ 50 μg C/l in supplies using ozonation in water treatment. Biofilm formation rate values were found to range from < 1 pg ATP/cm(2)/d in supplies using anaerobic ground water as the source. Increase of heterotrophic plate counts is limited at AOC values below 10 μg C/l. At BFR values below 10 pg ATP/cm(2)/d the risk of exceeding the guideline value for aeromonads (90 percentile < 200 c.f.u./100 ml) is less than 20%. Calculations based on the decrease of the AOC concentration observed in distributions systems confirm that very low concentrations of AOC can cause considerable biofilm formation on the pipe wall. The methods for assessing the biostability of drinking water combine with the assessment of the Biofilm Formation Potential of materials in contact with drinking water, thus providing a framework, the Unified Biofilm Approach, for evaluating the biostability of drinking water and materials.     

Efficacy of silver-releasing rubber for the prevention of Pseudomonas aeruginosa biofilm formation in water

Abstract

The aim of the present study was to evaluate the efficacy of Elastoguard? silver-releasing rubber in preventing Pseudomonas aeruginosa biofilm formation in water. Biofilm formation by P. aeruginosa under various conditions in an in vitro model system was compared for silver-releasing and conventional rubber. Under most conditions tested, the numbers of sessile cells attached to silver-releasing rubber were considerably lower with reference to conventional rubber, although the effect diminished with increasing volumes. The release of silver also resulted in a decrease in planktonic cells. By exposing both materials simultaneously to conditions for biofilm growth, it became obvious that the antibiofilm effect was due to a reduction in the number of planktonic cells, rather than to contact-dependent killing of sessile cells. The data demonstrate that the use of silver-releasing rubber reduces P. aeruginosa biofilm in water and reduces the number of planktonic cells present in the surrounding solution.     

李斯特菌生物被膜形成的调控

单核细胞增生李斯特菌(Listeria monocyiogenes)能引起人和动物脑膜炎、败血症、流产和单核细胞增多等症状,临床发病率在美国和欧洲等西方发达国家大约为2-8例/10万人,死亡率20％-30％或更高,被WHO列为关系食品卫生安全的重要病源细菌之一一[1-2].该菌能在多数固体表面形成生物被膜,在食品生产、加工、运输和保藏过程中,一旦发生细菌感染并形成生物被膜便难以将其彻底清除,严重威胁着食品卫生安全[3],但其生物被膜形成的具体分子机制尚不清楚[4].