Lethal and mutagenic action of solar radiation on model microbiological test systems

Lethal, mutagenic and recombonogenic action of the solar radiation on the model microorganisms--phage T4, bacteria Escherichia coli and ascomycet Aspergillus nidulans--has been studied. A considerable lethal effect of the solar radiation on phage T4 and E. coli was found. An increasing of mutation frequency in E. coli and A. nidulans by sunlight was also revealed. Recombinogenic action of solar radiation has been demonstrated in the experiments with diploid A. nidulans strains. It was shown that the excision and postreplication repair systems took part in recovery of damages induced by sunlight. An important role of ultra-violet region (280-320 nm) solar radiation in induction of lethal and mutagenic effects was demonstrated for all investigated microorganisms.     

Effects of solar radiation on bacterial and fungal density on aquatic plant detritus

1. The effects of solar radiation on bacterial and fungal growth on aquatic macrophyte detritus were studied in a microcosm experiment. Senescent leaves of Phragmites australis were incubated for 63 days in shallow water in the shade under photosynthetically active radiation (PAR) together with ultraviolet radiation, or under filters removing either ultraviolet B (UVB) or both UVB and ultraviolet A (UVA). 2. Bacterial abundance and bacterial ³H-leucine incorporation in the water were measured, together with α- and β-D-glucosidase activity. In addition, bacterial abundance and fungal biomass associated with the litter were measured. 3. The results indicate that both PAR and UVA affect the micro-organisms involved in the decomposition of leaf litter. The α/β-D-glucosidase activity ratio was less than one in irradiated and more than one in shaded microcosms, suggesting a change in the substrate dissolved organic matter composition towards more β- than α-glycosidic linkages as a result of solar radiation. 4. Microcosms receiving UVB displayed a significantly higher β-D-glucosidase activity than shaded microcosms, and those exposed to PAR or PAR + UVA, demonstrating the potential importance of UVB radiation. 5. The free-living bacteria tended to be dominated by filamentous forms in microcosms subject to solar radiation, especially PAR, and attached microbial communities showed a greater tendency to be dominated by bacteria in irradiated microcosms than in shaded microcosms.     

Persistence and differential survival of fecal indicator bacteria in subtropical waters and sediments

Fecal coliforms and enterococci are indicator organisms used worldwide to monitor water quality. These bacteria are used in microbial source tracking (MST) studies, which attempt to assess the contribution of various host species to fecal pollution in water. Ideally, all strains of a given indicator organism (IO) would experience equal persistence (maintenance of culturable populations) in water; however, some strains may have comparatively extended persistence outside the host, while others may persist very poorly in environmental waters. Assessment of the relative contribution of host species to fecal pollution would be confounded by differential persistence of strains. Here, freshwater and saltwater mesocosms, including sediments, were inoculated with dog feces, sewage, or contaminated soil and were incubated under conditions that included natural stressors such as microbial predators, radiation, and temperature fluctuations. Persistence of IOs was measured by decay rates (change in culturable counts over time). Decay rates were influenced by IO, inoculum, water type, sediment versus water column location, and Escherichia coli strain. Fecal coliform decay rates were significantly lower than those of enterococci in freshwater but were not significantly different in saltwater. IO persistence according to mesocosm treatment followed the trend: contaminated soil > wastewater > dog feces. E. coli ribotyping demonstrated that certain strains were more persistent than others in freshwater mesocosms, and the distribution of ribotypes sampled from mesocosm waters was dissimilar from the distribution in fecal material. These results have implications for the accuracy of MST methods, modeling of microbial populations in water, and efficacy of regulatory standards for protection of water quality.     

TRANSFORMATION AND ALLELOPATHY OF NATURAL DISSOLVED ORGANIC CARBON AND TANNIC ACID ARE AFFECTED BY SOLAR RADIATION AND BACTERIA1

The aim of this study was to test whether abiotic and biotic factors may affect allelopathic properties. Therefore, we investigated how solar radiation and bacteria influence allelopathic effects of the plant‐derived, polyphenolic tannic acid (TA) on microalgae. Using a block design, lake water samples with and without TA were exposed to solar radiation or kept in darkness with or without bacteria for 3 weeks. Dissolved organic carbon (DOC), specific size fractions of DOC analyzed by chromatography–organic carbon detection (LC‐OCD), and concentrations of total phenolic compounds (TPC) were measured to follow the fate of TA in lake water with natural DOC exposed to photolytic and microbial degradation. DOC and TPC decreased in dark‐incubated lake water with TA and bacteria indicating microbial degradation. In contrast, exposure to solar radiation of lake water with TA and bacteria did not decrease DOC. Chromatographic analyses documented an accumulation of DOC mean size fraction designated as humic substances (HS) in sunlit water samples with TA. The recalcitrance of the humic fraction indicates that photolytic degradation may contribute to a DOC less available for bacterial degradation. Subsequent growth tests with Desmodesmus armatus (Chodat) E. Hegewald showed low but reproducible difference in algal growth with lower algal growth rate cultured in photolytically and microbially degraded TA in lake water than cultured in respective dark treatments. This finding highlights the importance of photolytic processes and microbial degradation influencing allelopathic effects and may explain the high potential of allelochemicals for structuring the phytoplankton community composition in naturally illuminated surface waters.     

Decontamination of drinking water by direct heating in solar panels

A device was developed for direct heating of water by solar radiation in a flow-through system of copper pipes. An adjustable thermostat valve prevents water below the chosen temperature from being withdrawn. The results show that it is possible to eliminate coliform and thermotolerant coliform bacteria from naturally contaminated river water by heating to temperatures of 65 °C or above. Artificial additions of Salmonella typhimurium , Streptococcus faecalis and Escherichia coli to contaminated river water were also inactivated after heating to 65 °C and above. The total viable count could be reduced by a factor of 1000. The heat-resistant bacteria isolated from the Mlalakuva River (Tanzania) were spore-forming bacteria which exhibited greater heat resistance than commonly used test bacteria originating from countries with colder climates. To provide a good safety margin it is recommended that an outlet water temperature of 75 °C be used. At that temperature the daily production was about 50 l of decontaminated water per m² of solar panel, an amount that could be doubled by using a heat exchanger to recycle the heat.     

Solar and temporal effects on Escherichia coli concentration at a Lake Michigan swimming beach

Studies on solar inactivation of Escherichia coli in freshwater and in situ have been limited. At 63rd St. Beach, Chicago, Ill., factors influencing the daily periodicity of culturable E. coli, particularly insolation, were examined. Water samples for E. coli analysis were collected twice daily between April and September 2000 three times a week along five transects in two depths of water. Hydrometeorological conditions were continuously logged: UV radiation, total insolation, wind speed and direction, wave height, and relative lake level. On 10 days, transects were sampled hourly from 0700 to 1500 h. The effect of sunlight on E. coli inactivation was evaluated with dark and transparent in situ mesocosms and ambient lake water. For the study, the number of E. coli samples collected (n) was 2,676. During sunny days, E. coli counts decreased exponentially with day length and exposure to insolation, but on cloudy days, E. coli inactivation was diminished; the E. coli decay rate was strongly influenced by initial concentration. In situ experiments confirmed that insolation primarily inactivated E. coli; UV radiation only marginally affected E. coli concentration. The relationship between insolation and E. coli density is complicated by relative lake level, wave height, and turbidity, all of which are often products of wind vector. Continuous importation and nighttime replenishment of E. coli were evident. These findings (i) suggest that solar inactivation is an important mechanism for natural reduction of indicator bacteria in large freshwater bodies and (ii) have implications for management strategies of nontidal waters and the use of E. coli as an indicator organism.     

O3浓度升高和太阳辐射减弱对小麦根际土壤微生物功能多样性的影响

采用开顶箱(OTC)法和遮光网技术,设置100 n L/L臭氧熏气与3个辐射减弱梯度结合,模拟臭氧浓度升高和太阳辐射减弱的复合大气背景。用BIOLOG生态测试板,采用孔平均颜色变化率法(AWCD)测定冬小麦根际土壤微生物利用不同碳源的能力,计算微生物群落多样性指数,对不同碳源的利用率进行了主成分分析。两年试验结果显示,臭氧熏气与太阳辐射减弱复合作用,降低了土壤微生物对碳源的利用速度和利用总量;除了聚合物以外其它碳源利用率显著降低;对土壤微生物多样性没有直接的影响;对碳源降解的抑制效应大于增强的O3与减弱的太阳辐射两因素各自的单独作用。太阳辐射减弱20%,一定程度上增加了对聚合物类的分解。O3熏气条件下太阳辐射减弱,糖类、胺类代谢变异度较高,受环境影响较大。     

Detection of a Lactobacillus substance that inhibits Escherichia coli

Recent studies have shown that certain lactobacilli strains have the ability to interfere with the adherence and growth of uropathogenic bacteria. This interaction is believed to be important in the maintenance of a normal urogenital flora and in the prevention of infection in females. In the present study, Lactobacillus casei ssp. rhamnosus GR-1 and Lactobacillus acidophilus 76 were found to exert an inhibitory effect on pyelonephritogenic mutant Escherichia coli Hu 734 and E. coli ATCC 25922. The bioactivity of the inhibitor produced by strain GR-1 was retained under pH buffered conditions and was bactericidal. The bioactive substance was heat labile, not precipitated by up to 80% ammonium sulphate, and extractable in chloroform. The data indicated that the inhibitor is not lactic acid or hydrogen peroxide and has a molecular weight greater than 12,000-14,000. Human urine supported production of the inhibitor and reduced and delayed outgrowth of the E. coli. The ability of L. casei GR-1 and possibly other lactobacilli strains to produce inhibitors of uropathogenic bacteria may have clinical importance and significance in the microbial ecology of the urogenital tract.     

Influence of solar ultraviolet radiation on photosynthesis and motility of marine phytoplankton

Abstract: The effects of solar ultraviolet radiation (UV) on carbon uptake, oxygen evolution and motility of marine phytoplankton were investigated in coastal waters at Kristineberg Marine Research Station on the west coast of Sweden (58° 30'N, 11° 30'E). The mean irradiances at noon above the water surface during the investigation period were: photosynthetic active radiation (PAR, 400–700 nm) 1670 μmol m⁻² s⁻¹; ultraviolet-A radiation (UV-A, 320–400 nm) 35.9 W m⁻² and ultraviolet-B radiation (UV-B, 280–320 nm) 1.7 W m⁻². UV-B radiation was much more attenuated with depth in the water column than were PAR and UV-A radiation. UV-B radiation could not be detected at depths greater than 100–150 cm. Inhibition of carbon uptake by UV-A and UV-B in natural phytoplankton populations was greatest at 50 cm depth and the effects of UV-B were greater than those of UV-A. At depths greater than 50 cm there was almost no effect of ultraviolet radiation on carbon uptake. PAR, UV-A and UV-B decreased oxygen evolution by the dinoflagellate Prorocentrum minimum . Inhibition of oxygen evolution was greater after 4 h than 2 h but it was not possible to distinguish the negative effects of the different light regimes. The motility of P. minimum was not affected by PAR, UV-A and UV-B. The importance of exposure of phytoplankton to different light regimes before being exposed to natural solar radiation is discussed.     

Effect of physiological age on radiation resistance of some bacteria that are highly radiation resistant.

Physiological age-dependent variation in radiation resistance was studied for three bacteria that are highly radiation resistant: Micrococcus radiodurans, Micrococcus sp. isolate C-3, and Moraxella sp. isolate 4. Stationary-phase cultures of M. radiodurans and isolate C-3 were much more resistant to gamma radiation than were log-phase cultures. This pattern of relative resistance was reversed for isolate 4. Resistance of isolate 4 to UV light was also greater during log phase, although heat resistance and NaCl tolerance after heat stress were greater during stationary phase. Radiation-induced injury of isolate 4 compared with injury of Escherichia coli B suggested that the injury process, as well as the lethal process, was affected by growth phase. The hypothesis that growth rate affects radiation resistance was tested, and results were interpreted in light of the probable confounding effect of methods used to alter growth rates of bacteria. These results indicate that dose-response experiments should be designed to measure survival during the most resistant growth phase of the organism under study. This timing is particularly important when extrapolations of survival results might be made to potential irradiation processes for foods.     

Effect of physiological age on radiation resistance of some bacteria that are highly radiation resistant

Physiological age-dependent variation in radiation resistance was studied for three bacteria that are highly radiation resistant: Micrococcus radiodurans, Micrococcus sp. isolate C-3, and Moraxella sp. isolate 4. Stationary-phase cultures of M. radiodurans and isolate C-3 were much more resistant to gamma radiation than were log-phase cultures. This pattern of relative resistance was reversed for isolate 4. Resistance of isolate 4 to UV light was also greater during log phase, although heat resistance and NaCl tolerance after heat stress were greater during stationary phase. Radiation-induced injury of isolate 4 compared with injury of Escherichia coli B suggested that the injury process, as well as the lethal process, was affected by growth phase. The hypothesis that growth rate affects radiation resistance was tested, and results were interpreted in light of the probable confounding effect of methods used to alter growth rates of bacteria. These results indicate that dose-response experiments should be designed to measure survival during the most resistant growth phase of the organism under study. This timing is particularly important when extrapolations of survival results might be made to potential irradiation processes for foods.     

Modulation of the bacterial response to spectral solar radiation by algae and limiting nutrients

SUMMARY 1. The response of bacterial production (measured as [³H]TdR incorporation rate) to spectral solar radiation was quantified experimentally in an oligotrophic high-mountain lake over 2 years. Bacterial responses were consistent: ultraviolet-B (UVB) was harmful, whereas ultraviolet-A (UVA) + photosynthetically active radiation (PAR) and PAR enhanced bacterial activity. Full sunlight exerted a net stimulatory effect on bacterial activity in mid-summer but a net inhibitory effect towards the end of the ice-free period.
2. Experiments were undertaken to examine whether the bacterial response pattern depended on the presence of algae and/or was modulated by the availability of a limiting inorganic nutrient (phosphorus, P). In the absence of algae, [³H]TdR incorporation rates were significantly lower than when algae were present under all light treatments, and the consistent bacterial response was lost. This suggests that the bacterial response to spectral solar radiation depends on fresh-C released from algae, which determines the net stimulatory outcome of damage and repair in mid-summer.
3. In the absence of algae, UVB radiation inhibited bacteria when they were strongly P-deficient (mean values of N : P ratio: 46.1), whereas it exerted no direct effect on bacterial activity when they were not P-limited.
4. P-enrichment of lake water markedly altered the response of bacteria to spectral solar radiation at the end of ice-free period, when bacteria were strongly P-deficient. Phosphorus enrichment suppressed the inhibitory effect of full sunlight that was observed in October, both in whole lake water (i.e. including algae) and in the absence of algae. This indicates that the bacterial P-deficiency, measured as the cellular N : P ratio, was partly responsible for the net inhibitory effect of full sunlight, implying a high bacterial vulnerability to UVB.     

RecA Expression in Response to Solar UVR in the Marine Bacterium Vibrio natriegens

Solar ultraviolet radiation may produce daily stress on marine and estuarine communities as cells are damaged and repair that damage. Reduction in the earth's stratospheric ozone layer has increased awareness of the potential effects that ultraviolet radiation may have in the environment, including how marine bacteria respond to changes in solar radiation. We examined the use of the bacterial RecA protein as an indicator of the potential of bacteria to repair DNA damage caused by solar UV irradiation using the marine bacterium Vibrio natriegens as a model. RecA is universally present in bacteria and is a regulator protein for the so-called Dark Repair Systems, which include excision repair, postreplication recombinational repair, and mutagenic or SOS repair. Solar UVB and UVA both reduced V. natriegens viability in seawater microcosms. After exposure to unfiltered solar radiation or radiation in which UVB was blocked, survival dropped below 1%, whereas visible light from which UVA and UVB had been filtered had no effect on survival. Using a RecA-specific antibody for detection, RecA protein was induced by solar radiation in a diel pattern in marine microcosms conducted in the Gulf of Mexico. Peak induction was observed at dusk each day. Although RecA expression was correlated with the formation of UVB-induced cyclobutyl pyrimidine dimers, longer wavelength UVA radiation also induced recA gene expression. Our results demonstrate that RecA-regulated, light-independent repair is an important component in the ability of marine bacteria to survive exposure to solar ultraviolet radiation and that RecA expression is a useful monitor of bacterial repair after exposure to solar UVR.     

粤北山地常绿阔叶林自然干扰后冠层结构与林下光照动态

以粤北车八岭2008年受冰灾破坏的山地常绿阔叶林为研究对象,设置2 hm2固定样地开展连续3a(2008—2010年)的群落调查,并采用半球面影像技术(Hemispherical photography)获取冠层结构和林下光照指标,分析灾后森林演替过程中冠层结构和林下光照的动态。研究发现:1)灾后森林恢复过程中,样地林下光照(直射光、散射光和总光照)均随林冠开度的减少、叶面积指数的增加而减少;2)从2008到2010年,各年度冠层结构和林下光照的差异均极显著(P<0.0001),但年间差异程度有逐年减少的趋势;3)灾后森林恢复前3a,林下直射光对林下总光照的贡献大于散射光,其时空波动性也大于散射光;4)林冠开度对冠层结构的反映程度比叶面积指数高,冠层结构对林下散射光的影响比对直射光大。灾后林木先是迅速生长然后生长速度缓慢下来并逐渐稳定,随森林逐渐郁闭林下光照也随之减少,其中林冠开度用于评价冠层结构动态的效果更佳,林下直射光比散射光的时空变化更复杂。     

Effects of Experimental Lead Pollution on the Microbial Communities Associated with <Emphasis Type="Italic">Sphagnum fallax</Emphasis> (Bryophyta)

Ecotoxicological studies usually focus on single microbial species under controlled conditions. As a result, little is known about the responses of different microbial functional groups or individual species to stresses. In an aim to assess the response of complex microbial communities to pollution in their natural habitat, we studied the effect of a simulated lead pollution on the microbial community (bacteria, cyanobacteria, protists, fungi, and micrometazoa) living on Sphagnum fallax. Mosses were grown in the laboratory with 0 (control), 625, and 2,500 microg L(-1) of Pb(2+) diluted in a standard nutrient solution and were sampled after 0, 6, 12, and 20 weeks. The biomasses of bacteria, microalgae, testate amoebae, and ciliates were dramatically and significantly decreased in both Pb addition treatments after 6, 12, and 20 weeks in comparison with the control. The biomass of cyanobacteria declined after 6 and 12 weeks in the highest Pb treatment. The biomasses of fungi, rotifers, and nematodes decreased along the duration of the experiment but were not significantly affected by lead addition. Consequently, the total microbial biomass was lower for both Pb addition treatments after 12 and 20 weeks than in the controls. The community structure was strongly modified due to changes in the densities of testate amoebae and ciliates, whereas the relative contribution of bacteria to the microbial biomass was stable. Differences in responses among the microbial groups suggest changes in the trophic links among them. The correlation between the biomass of bacteria and that of ciliates or testate amoebae increased with increasing Pb loading. We interpret this result as an effect on the grazing pathways of these predators and by the Pb effect on other potential prey (i.e., smaller protists). The community approach used here complements classical ecotoxicological studies by providing clues to the complex effect of pollutant-affecting organisms both directly and indirectly through trophic effects and could potentially find applications for pollution monitoring.     

An in situ enclosure experiment to test the solar UVB impact on plankton in a high-altitude mountain lake. II. Effects on the microbial food web

We studied the impact of ambient levels of solar ultraviolet B (UVB) radiation on the planktonic microbial food web (viruses, heterotrophic bacteria, heterotrophic nanoflagellates and ciliates) of a high-mountain lake (2417 m above sea level) under in situ conditions for 16 days. Enclosures of 1 m³ receiving either the full sunlight spectrum or sunlight without UVB radiation were suspended at the lake surface. We found that the abundance of heterotrophic nanoflagellates was always lower in the +UVB treatment than in the -UVB one. In addition, bacterial consumption, measured by the disappearance of fluorescently labelled bacteria, was significantly (P < 0.05) reduced in the +UVB treatment. The abundance of non-filamentous bacteria (<10 m long) was also lower in the +UVB treatment, suggesting a direct effect of UVB on their growth. This was supported by the significantly (P < 0.05) lower cell-specific activity ([³H])thymidine incorporation) found on the fifth day of the experiment. In contrast, UVB radiation had no effect on filamentous bacteria (>1 m long), which represented only a small fraction of the total abundance (<4%), but up to 70% of the total bacterial biovolume. Ciliates, mainly Urotricha pelagica and Urotricha furcata, were less impacted by UVB radiation, although the net growth rate during the first week of the experiment was lower in the +UVB treatment than in the -UVB one (0.22 and 0.39 day^-1, respectively). The abundance of virus-like particles during the first week of the experiment was higher in the -UVB treatment. After reaching the maximum value for the interaction viruses x bacteria, viral abundance decreased dramatically (by 85%) in both treatments with a decay rate of 0.017 h^-1. This study illustrates the complexity in assessing the impact of UVB radiation when more than one trophic level is considered and indicates the existence of different sensitivity to UVB radiation among components of the microbial food web.      

Effect of UV Radiation on the Bacterivory of a Heterotrophic Nanoflagellate

The effects of UV-B radiation on the heterotrophic nanoflagellate Bodo saltans (Kinetoplastida) were examined under controlled conditions with artificial UV sources and also under natural solar radiation in an oligotrophic lake. In both types of experiments, the characteristic elongated cell morphology of this flagellate changed into a spherical one. This effect was due to UV-B but also to UV-A radiation, and after 4 h of exposure at 0.5 m of depth, 99% (UV-B plus UV-A plus photosynthetically active radiation) and 69% of the cells (UV-A plus photosynthetically active radiation) were spherical. At 6 m of depth where only 10% of the UV-B (305 nm) at the surface was measured, no significant effect was observed. The spherical cells were nonmotile, but before the morphological change took place, the swimming speed was ca. 3.5 times lower in the plus-UV-B treatment. The negative relation between the abundance of spherical cells and the average ingestion of fluorescently labeled bacteria per cell indicates that these cells are not able to feed upon bacteria. In bacterivory experiments lasting for 6 h, the total number of grazed bacteria was up to 70% lower in the plus-UV-B treatment than in the control without UV-B. This resulted in a positive feedback between UV-B and bacterial growth. The high sensitivity of B. saltans to solar UV-B and UV-A radiation strongly reduces its ability to live near the surface at times of high UV radiation.     

OLAND生物脱氮系统中硝化菌群16S rDNA的DGGE分析

为了考察生物脱氮系统中硝化菌群（氨氧化菌和亚硝酸氧化菌）的种群多样性及硝化菌群随溶解氧降低的种群变化规律,并建立一套行之有效的用于自养生物脱氮系统中功能微生物菌群的快速分子检测技术,采用DGGE（变性梯度凝胶电泳）分子检测技术对硝化菌群的16SrDNA的特异性PCR扩增产物进行了分析,结果表明：OLAND生物脱氮系统中氨氧化菌和亚硝酸氧化菌随溶解氧的降低表现出了不同的种群变化规律,氨氧化菌种群多样性受溶解氧的影响非常大,而非亚硝酸氧化菌的种群多样性比较单一,且不受溶解氧的影响。结合FISH（全细胞荧光原位杂交）分析结果表明,在OLAND限氧稳定运行后期,亚硝化单胞菌属（Nitrosomonas）是主要的氨氧化菌,占OLAND限氧亚硝化阶段反应器中总细菌数的72.5%左右。     

Nitrogen Fixation in Microbial Mat and Stromatolite Communities from Cuatro Cienegas,Mexico

Nitrogen fixation (nitrogenase activity, NA) of a microbial mat and a living stromatolite from Cuatro Cienegas, Mexico, was examined over spring, summer, and winter of 2004. The goal of the study was to characterize the diazotrophic community through molecular analysis of the nifH gene and using inhibitors of sulfate reduction and oxygenic and anoxygenic photosynthesis. We also evaluated the role of ultraviolet radiation on the diazotrophic activity of the microbial communities. Both microbial communities showed patterns of NA with maximum rates during the day that decreased significantly with 3-3,4-dichlorophenyl-1′,1′-dimethylurea, suggesting the potential importance of heterocystous cyanobacteria. There is also evidence of NA by sulfur-reducing bacteria in both microbial communities suggested by the negative effect exerted by the addition of sodium molybdate. Elimination of infrared and ultraviolet radiation had no effect on NA. Both microbial communities had nifH sequences that related to group I, including cyanobacteria and purple sulfur and nonsulfur bacteria, as well as group II nitrogenases, including sulfur reducing and green sulfur bacteria.     

Role of bacteria and protozoa in the removal of Escherichia coli from estuarine waters.

The removal of Escherichia coli from estuarine water was investigated. The survival of E. coli was dependent on the presence of protozoan predators and not on the presence of lytic bacteria. When indigenous protozoa were removed from estuarine water by filtration, the destruction of coliform populations was negligible. In studies designed to prevent the growth of indigenous bacterial populations without affecting protozoan populations, coliform destruction was significant.