Growth ofEnteromorpha linza in sewage effluent and sewage effluent-seawater mixtures

Enteromorpha linza (L.) J. Ag. was grown in full strength sewage effluent, various combinations of sewage effluent and seawater, and in natural seawater. It was found that full strength sewage effluent with a salinity of 14 supported best growth of the alga. After a 12 day cultivation period, growth ofE. linza in full strength sewage effluent and 75% sewage effluent- seawater mixture showed 3.5-fold and 2-fold increase in fresh weight over that grown in natural seawater; respectively. Uptake of PO _inf4 ^sup3– -P, NH₃-N and NO _inf3 ^sup– -N by cells ofE. linza was extremely efficient in all tested media. Data obtained from the experiments indicated that inorganic nitrogen rather than phosphorus was the limiting nutrient factor for growth ofE. linza in full strength sewage effluent and in other sewage effluent- seawater mixtures. NH₃-N at concentrations above 4.5 ppm was found to inhibit uptake of NO _inf3 ^sup– -N in the same culture medium by the algal cells. The fact that sewage grownE. linza contained comparatively much higher protein content (30.2% dry weight) than that grown in natural seawater (12.5% dry weight) leads to the conclusion that sewage grownE. linza could serve as an economically feasible feed for livestock in Hong Kong where the sewage is characterized by having a salinity of approximately 14. It is proposed that this multicellular green alga is a suitable algal species to serve the dual function of wastewater purification through the production of algal protein from sewage effluent having high salinities.     

Microbial transformation of nitroaromatic compounds in sewage effluent

The transformation of mono- and dinitroaromatic compounds was measured in sewage effluent maintained under aerobic or anaerobic conditions. Most of the nitrobenzene, 3- and 4-nitrobenzoic acids, and 3- and 4-nitrotoluenes and much of the 1,2- and 1,3-dinitrobenzenes disappeared both in the presence and absence of oxygen. Under anaerobiosis, 2,6-dinitrotoluene and 3,5-dinitrobenzoic acid disappeared slowly, but no loss was evident in 28 days in aerated sewage. Aromatic amines did not accumulate during the aerobic decomposition of the mononitro compounds. They did appear in nonsterile, but not in sterile, sewage incubated aerobically with the dinitro compounds and anaerobically with all the chemicals. Analysis by gas chromatography and combined gas chromatography-mass spectrometry showed that aniline was formed from nitrobenzene, toluidine was formed from 3- and 4-nitrotoluenes, and aminobenzoic acid was formed from 3- and 4-nitrobenzoic acids under anaerobiosis, and that nitroaniline was formed from 1,2- and 1,3-dinitrobenzenes, aminonitrotoluene resulted from 2,6-dinitrotoluene, and aminonitrobenzoic acid was a product of 3,5-dinitrobenzoic acid under both conditions. The isomeric forms of the metabolites were not established. Aniline, 4-toluidine, and 4-aminobenzoic acid added to sewage disappeared from aerated nonsterile, but not from sterile, sewage or sewage in the absence of oxygen. 2-Nitroaniline, 2-amino-3-nitrotoluene, and 2-amino-5-nitrobenzoic acid added to sewage persisted for at least 60 days in aerobic or anaerobic conditions. Gas chromatographic and gas chromatographic-mass spectrometric analyses demonstrated that acetanilide and 2-methylquinoline were formed from aniline, 4-methylformanilide and 4-methylacetanilide were formed from 4-toluidine, 2-methylbenzimidazole was a product of 2-nitroaniline, and unidentified benzimidazoles were formed from 2-amino-3-nitrotoluene in the absence of oxygen, and that 2-nitroacetanilide and 2-methyl-6-nitroacetanilide were formed from 2-nitroaniline and 2-amino-3-nitrotoluene, respectively, in the presence or absence of oxygen. It is suggested that the transformations of widely used nitroaromatic compounds should be further studied because of the persistence and possible toxicity of products of their metabolism.     

Growth of water hyacinths in treated sewage effluent

Two thousand plants of the water hyacinth,Eichornia crassipes Solms., were introduced on April 11, 1971, into a series of five ponds, each 5000 sq. ft. in area and 2.6 ft. deep. Treated waste water effluent from the Ames sewage treatment plant filled the ponds and was added to pond 1 at 127 gallons per minute. By growth and vegetative reproduction, these plants increased to more than 500,000, and all five ponds were covered completely by July 26. On that date, the extrapolated estimate of total wet weight was 287 U.S. tons/ acre (645 metric tons/hectare; 64500g/m²). The estimate of oven dry weight was 13.2 U.S. tons/acre (29.7 metric tons/hectare; 2970g/m²). Ammonia and nitrate disappeared rapidly from the pond water, and phosphate concentrations were lowered appreciably. Evapotranspiration and seepage accounted for water losses of more than 0.5 inches per day. The potential economic values of this plant and its possible use in tertiary treatment to reduce N and P components in waste waters are discussed briefly.     

Microbial transformation of nitroaromatic compounds in sewage effluent

[This corrects the article on p. 1239 in vol. 45.].     

Metabolic removal of phosphate from sewage effluent

Eutrophication, or fertilization, has become a major water pollution problem associated with the discharge of mineral-rich sewage eflluent. A metabolic process to remove dissolved phosphate from sewage through the action of sewage microorganisms is under development. The process, unlike other proposed solutions to the problem, would not require tertiary treatment of the sewage. Laboratory studies have produced promising data. Early reports from municipal sewage treatment plants confirm the expectation that the process may be feasible for widespread use.     

Occurrence of Cryptosporidium oocysts and Giardia cysts in sewage in Norway

Samples of sewage influent from 40 sewage treatment works (STW) throughout Norway were examined for Cryptosporidium oocysts and Giardia duodenalis cysts. Both parasites were detected frequently (80% of STW were Cryptosporidium positive; 93% of STW were Giardia positive) and at maximum concentrations of > 20,000 parasites/liter. The data suggest giardiasis is more widespread, and/or occurs with greater infection intensity, than cryptosporidiosis in Norway. STW serving higher person equivalents were more likely to be positive and had higher parasite concentrations. Parasite concentrations were used to estimate the proportion of contributing populations that could be clinically infected. For Cryptosporidium, the highest estimates were up to 5 per 100,000 individuals for two populations in eastern Norway. For Giardia, the highest estimate was 40 infected per 100,000 persons (approximately five times the usual national annual average) contributing to an STW in western Norway. As this population experienced a large waterborne giardiasis outbreak 6 months after sampling, it can be speculated that regular challenge with Giardia may occur here. Most Giardia isolates in sewage influent were assemblage A, although some assemblage B isolates were detected. There was substantial heterogeneity, but most samples contained isolates similar to genotype A3. Removal efficiencies at two STW with secondary treatment processes were estimated to be approximately 50% for Cryptosporidium and > 80% for Giardia. An STW with minimal treatment had negligible removal of both parasites. Many STW in Norway have minimal treatment and discharge effluent into rivers and lakes, thus, risk of contamination of water courses by Cryptosporidium and Giardia is considerable.     

Chlorination of indicator bacteria and viruses in primary sewage effluent

Wastewater disinfection is used in many countries for reducing fecal coliform levels in effluents. Disinfection is therefore frequently used to improve recreational bathing waters which do not comply with microbiological standards. It is unknown whether human enteric viruses (which are responsible for waterborne disease) are simultaneously inactivated alongside fecal coliforms. This laboratory study focused on the chlorination of primary treated effluent with three doses (8, 16, and 30 mg/liter) of free chlorine as sodium hypochlorite. Seeding experiments showed that inactivation (>5 log(10) units) of Escherichia coli and Enterococcus faecalis was rapid and complete but that there was poor inactivation (0.2 to 1.0 log(10) unit) of F(+)-specific RNA (FRNA) bacteriophage (MS2) (a potential virus indicator) at all three doses. However, seeded poliovirus was significantly more susceptible (2.8 log(10) units) to inactivation by chlorine than was the FRNA bacteriophage. To ensure that these results were not artifacts of the seeding process, comparisons were made between inactivation rates of laboratory-seeded organisms in sterilized sewage and inactivation rates of organisms occurring naturally in sewage. Multifactorial analysis of variance showed that there was no significant difference (P > 0.05) between the inactivation rates for seeded and naturally occurring FRNA bacteriophage. However, laboratory-grown poliovirus was inactivated much more rapidly than were naturally occurring, indigenous enteroviruses (P < 0.001). This may reflect differences in the way indigenous virus is presented to the disinfectant. Inactivation rates for indigenous enteroviruses were quite similar to those seen for FRNA bacteriophage at lower doses of chlorine. These results have significance for the effectiveness of chlorination as a sewage treatment process, particularly where virus contamination is of concern, and suggest that FRNA bacteriophage would be an appropriate indicator of such viral inactivation under field conditions.     

Detection of Giardia in Sewage Effluent1

Giardia sp. cysts were found at levels of 4,000–450,000/378,500 liters (100,000 gallons) in sewage effluents from three of seven sewage treatment plants in Sangamon County, Illinois, in June, July, and August 1981. Effluent from the positive plants is discharged into Lake Springfield (the present source of the city of Springfield's water supply) or the Sangamon River.     

Response of pondcypress growth rates to sewage effluent application

Cores were collected from dominant pondcypress trees growing in a swamp that had received sewage effluent for 7 yr and a nearby control swamp to determine the combined effects of changes in nutrient supply and hydrologic regime on tree growth. The cores were used to measure two indices of tree growth: basal area increment (BAI) and relative basal area increment (RBAI, which accounts for differences in growth due to the size of teh tree) between 1970–1983 while one swamp remained untreated and the other received weekly additions of sewage effluent from 1974–1981. Throughout the whole period, the mean BAI and RBAI of pond-cypress trees in the untreated swamp remained unchanged, ranging between 5.55–6.38 cm² yr^–1 and 1.09–1.27% yr^–1, respectively. In contrast, trees in the treated swamp increased their BAI approximately two-fold from 7.40 cm² yr^–1 prior to treatment to 14.83 cm² yr^–1 after the onset of treatment and maintained this rate of growth in the 2 yr period after cessation of treatment. Relative basal area increment showed a similar response, but the proportional increase due to treatment was less (1.5-fold factor) than for BAI. The response of pondcypress trees to the sewage effluent differed depending upon whether the trees were located in the deep or shallow water zones. Trees in the deep zone of the treated swamp had lower BAIs and RBAIs than those in the shallow zone during the treatment period, whereas in pre- and post-treatment periods growth indices were equal in both zones. No significant differences in growth between deep and shallow zones were observed during all three time periods in the control swamp.     

An assessment of a natural wetland receiving sewage effluent

An assessment of the performance of a wetland dominated by opportunistic weeds in removing nutrients from a secondary sewage effluent was carried out at Thredbo in Kosciusko National Park. Water quality sampling of the inflow and outflow showed a reduction in turbidity, pH and conductivity with passage through the wetland. Dissolved oxygen levels also decreased. In summer 1982, the phosphorus toad of 1.8 kg P day^-1 was reduced by 44% as wastewater moved through the wetland, and the nitrogen load of 6.4 kg N day^-1 by 65%. Winter retentions were 10% and 14% of inflowing loads, respectively. In 1983 there was a net release of phosphorus and reduced retention of nitrogen. This was regarded as being a result of physical disturbance of the wetland. Vegetation downstream of the effluent inflow was dominated by the opportunistic weeds Epilobium sarmantaceum and Rumex crispus. Upstream, Carex gaudichaudiana and Baeckea gunniana dominated. Marked seasonal changes in vigour and biomass were recorded. Plants, but not sediments, downstream of the inflow contained more nutrients than those upstream. Seed bank analysis showed more seeds downstream than upstream. A moist treatment promoted more seed germination than a flooded treatment. E. sarmantaceum and to a lesser extent R. crispus dominated in each treatment. In an effort to promote greater water retention and replacement of the weed species, channels in the wetland have been blocked with gravel and planted with Phragmites australis and Schoenoplectus validus.     

Detection of Giardia lamblia by immunofluorescence

High-titer immune sera to cysts of Giardia lamblia, produced in guinea pigs, were labeled with fluorescein isothiocyanate. The resulting conjugates were used to detect G. lamblia in stool specimens by fluorescence microscopy. The sera also reacted with cysts of Chilomastix mesnili, but the two organisms could be differentiated by their size.     

Simultaneous detection of Cryptosporidium parvum and Giardia in sewage sludge by IC-PCR

AIMS: The aim of this study was to develop a method based on immunomagnetic capture and polymerase chain reaction (IC-PCR assay) for detection of Cryptosporidium parvum and Giardia intestinalis in sewage sludge. METHODS AND RESULTS: The detection limit of the IC-PCR assay for both organisms was 625 oocysts and cysts ml(-1). By hybridization of PCR products the sensitivity could be increased to 125 oocysts and cysts ml(-1). Forty-four sludge samples from 12 wastewater treatment plants were examined. The samples positive for Giardia (9 out of 44) were from eight wastewater plants and the C. parvum genotype 2 samples (3 out of 44) originated from different sewage works. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: IC-PCR offers the possibility to distinguish between Cryptosporidium and Giardia genotypes. This assay can be used to monitor the presence of these organisms in a community and to determine contamination of sludge used as soil amendment.     

Effects of sewage waste stabilization lagoon effluent on stream invertebrates

The ecological impact of discharge to streams of domestic sewage waste stabilization lagoon effluent was investigated. Benthic invertebrates were compared upstream and downstream of discharges to eight New Zealand streams where effluent dilution ranged from 6- to 484-fold. The percentage of common invertebrate taxa whose density changed significantly (ANOVA, P<0.05) downstream declined in proportion with the log of the effluent dilution (r=-0.87) and increased with downstream increase in benthic respiration (r=0.91) and several intercorrelated indicators of organic enrichment (log biochemical oxygen demand, r=0.91; log suspended solids (SS), r=0.84). However, these changes in invertebrate densities did not always reflect degraded community structure. The nature and direction of changes suggests a subsidy-stress gradient of responses. Increases in SS of > 4 g m^-3 were associated with significant changes in density of > 50 percent of the common taxa and > 50 percent reduction of the densities of the sensitive Ephemeroptera, Plecoptera, and Trichoptera (EPT). However, EPT densities increased by up to 50 percent at lower organic solids loadings. No general relationships were found between relative densities of functional feeding groups and metabolic or water quality variables. These findings confirm that early warning of enrichment stress is more easily seen at the species level than at the functional level.     

Virus movement in soil columns flooded with secondary sewage effluent.

Secondary sewage effluent containing about 3 X 10(4) plaque-forming units of polio virus type 1 (LSc) per ml was passed through columns 250 cm in length packed with calcareous sand from an area in the Salt River bed used for ground-water recharge of secondary sewage effluent. Viruses were not detected in 1-ml samples extracted from the columns below the 160-cm level. However, viruses were detected in 5 of 43 100-ml samples of the column drainage water. Most of the viruses were adsorbed in the top 5 cm of soil. Virus removal was not affected by the infiltration rate, which varied between 15 and 55 cm/day. Flooding a column continuosly for 27 days with the sewage water virus mixture did not saturate the top few centimeters of soil with viruses and did not seem to affect virus movement. Flooding with deionized water caused virus desorption from the soil and increased their movement through the columns. Adding CaCl2 to the deionized water prevented most of the virus desorption. Adding a pulse of deionized water followed by sewage water started a virus front moving through the columns, but the viruses were readsorbed and none was detected in outflow samples. Drying the soil for 1 day between applying the virus and flooding with deionized water greatly reduced desorption, and drying for 5 days prevented desorption. Large reductions (99.99% or more) of virus would be expected after passage of secondary sewage effluent through 250 cm of the calcareous sand similar to that used in our laboratory columns unless heavy rains fell within 1 day after the application of sewage stopped. Such virus movement could be minimized by the proper management of flooding and drying cycles.     

A new sponge tray bioreactor in primary treated sewage effluent treatment

The new attached growth sponge tray bioreactor (STB) was evaluated at different operating conditions for removing organics and nutrients from primary treated sewage effluent. This STB was also assessed when using as a pre-treatment prior to micro-filtration (MF) for reducing membrane fouling. At a short hydraulic retention time (HRT) of 40 min, the STB could remove up to 92% of DOC and 40-56% of T-N and T-P at an organic loading rate (OLR) of 2.4 kg COD/m³ sponge day. This OLR is the best for the STB as compared to the OLRs of 0.6, 1.2 and 3.6 kg COD/m³ sponge day. At 28 mL/min of flow velocity (FV), STB achieved the highest efficiencies with 92% of DOC, 87.4% of T-P, and 54.8% of T-N removal. Finally, at the optimal OLR and FV, the STB could remove almost 90% of organic and nutrient, significantly reduce membrane fouling with HRT of only 120 min.     

Virus movement in soil columns flooded with secondary sewage effluent.

Secondary sewage effluent containing about 3 X 10(4) plaque-forming units of polio virus type 1 (LSc) per ml was passed through columns 250 cm in length packed with calcareous sand from an area in the Salt River bed used for ground-water recharge of secondary sewage effluent. Viruses were not detected in 1-ml samples extracted from the columns below the 160-cm level. However, viruses were detected in 5 of 43 100-ml samples of the column drainage water. Most of the viruses were adsorbed in the top 5 cm of soil. Virus removal was not affected by the infiltration rate, which varied between 15 and 55 cm/day. Flooding a column continuosly for 27 days with the sewage water virus mixture did not saturate the top few centimeters of soil with viruses and did not seem to affect virus movement. Flooding with deionized water caused virus desorption from the soil and increased their movement through the columns. Adding CaCl2 to the deionized water prevented most of the virus desorption. Adding a pulse of deionized water followed by sewage water started a virus front moving through the columns, but the viruses were readsorbed and none was detected in outflow samples. Drying the soil for 1 day between applying the virus and flooding with deionized water greatly reduced desorption, and drying for 5 days prevented desorption. Large reductions (99.99% or more) of virus would be expected after passage of secondary sewage effluent through 250 cm of the calcareous sand similar to that used in our laboratory columns unless heavy rains fell within 1 day after the application of sewage stopped. Such virus movement could be minimized by the proper management of flooding and drying cycles.     

Denitrification in a South Louisiana wetland forest receiving treated sewage effluent

Although denitrification has the potential to reduce nitrate (NO ₃ ^– ) pollution of surface waters, the quantification of denitrification rates is complex because it requires differentiation from other mechanisms and is highly variable in both space and time. This study first measured potential denitrification rates at a wetland forest site in south Louisiana before receipt of secondary wastewater effluent, and then, following 30 months of effluent application, landscape gradients of dissolved nitrate (NO ₃ ^– ) and nitrous oxide (N₂O) were measured. A computer model was developed to quantify N transformations. Floodwater NO ₃ ^– and N₂O concentrations were higher in the forest receiving effluent than in the adjacent control forest. Denitrification rates of NO ₃ ^– -amended soil cores ranged from 0.03–0.45 g N m^–2 d^–1 with an overall mean of 0.10 g N m^–2 d^–1. Effluent N is being applied at a rate of approximately 0.034 g N m^–2 d^–1, with approximately 95% disappearing along a 1 km transect. In the treatment forest, floodwater NO ₃ ^– concentrations decreased from 1000 M at the inflow point to 50 M along the 1 km transect. Nitrous oxide concentrations increased from 0.25 M to 1.2 M within the first 100 m, but decreased to 0.1 M over the next 900 m. The initial increase in N₂O was presumably a result ofin situ denitrification. Model analyses indicated that denitrification was directly associated with nitrification and was limited by the availability of NO ₃ ^– produced by nitrification. Due to different redox potential optima, coupling of nitrification and denitrification was a function of a balance of environmental conditions that was moderately favorable to both processes. N removal efficiency was largely dependent on the proportion of effluent NH ₄ ⁺ to NO ₃ ^– . When NH ₄ ⁺ /NO ₃ ^– was 1, average N removal efficiency ranged from 95–100%, but ratios that were >1 reduced average efficiencies to as low as 57%. Actual effluent NH ₄ ⁺ /NO ₃ ^– loading ratios at this site are approximately 0.2 and are consistently <1.     

Detection of Giardia lamblia by immunofluorescence.

High-titer immune sera to cysts of Giardia lamblia, produced in guinea pigs, were labeled with fluorescein isothiocyanate. The resulting conjugates were used to detect G. lamblia in stool specimens by fluorescence microscopy. The sera also reacted with cysts of Chilomastix mesnili, but the two organisms could be differentiated by their size.     

Identification of co-mutagenic chlorinated harmans in final effluent from a sewage treatment plant

Harman (1-methyl-9H-pyrido[3,4-b]indole) reacted readily with sodium hypochlorite in an aqueous medium to give the mono-chlorinated derivatives, which reportedly have greater co-mutagenic activity than harman in the presence of o-toluidine toward Salmonella typhimurium TA 98 with S9 mix. Mono-chlorinated harmans were detected by concentration using blue rayon (BR) and GC/MS analysis in the final effluent from a sewage treatment plant in Shizuoka, Japan. The amounts adsorbed for 24h were 1-45ng/gBR for mono-chlorinated harman and 110-730ng/gBR for harman.     

Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage

The protozoan pathogens Giardia lamblia and Cryptosporidium parvum are major causes of waterborne enteric disease throughout the world. Improved detection methods that are very sensitive and rapid are urgently needed. This is especially the case for analysis of environmental water samples in which the densities of Giardia and Cryptosporidium are very low. Primers and TaqMan probes based on the beta-giardin gene of G. lamblia and the COWP gene of C. parvum were developed and used to detect DNA concentrations over a range of 7 orders of magnitude. It was possible to detect DNA to the equivalent of a single cyst of G. lamblia and one oocyst of C. parvum. A multiplex real-time PCR (qPCR) assay for simultaneous detection of G. lamblia and C. parvum resulted in comparable levels of detection. Comparison of DNA extraction methodologies to maximize DNA yield from cysts and oocysts determined that a combination of freeze-thaw, sonication, and purification using the DNeasy kit (Qiagen) provided a highly efficient method. Sampling of four environmental water bodies revealed variation in qPCR inhibitors in 2-liter concentrates. A methodology for dealing with qPCR inhibitors that involved the use of Chelex 100 and PVP 360 was developed. It was possible to detect and quantify G. lamblia in sewage using qPCR when applying the procedure for extraction of DNA from 1-liter sewage samples. Numbers obtained from the qPCR assay were comparable to those obtained with immunofluorescence microscopy. The qPCR analysis revealed both assemblage A and assemblage B genotypes of G. lamblia in the sewage. No Cryptosporidium was detected in these samples by either method.