Removal efficiency and toxicity reduction of 4-chlorophenol with physical, chemical and biochemical methods.

Chlorophenols are well-known priority pollutants and many different treatments have been assessed to facilitate their removal from industrial wastewater. However, an absolute and optimum solution still has to be practically implemented in an industrial setting. In this work, a series ofphysical, chemical and biochemical treatments have been systematically tested for the removal of 4-chlorophenol, and their results have been compared in order to determine the most effective treatment based on removal efficiency and residual by-product formation. Chemical treatments based on advanced oxidation processes (AOP) produced the best results on rate and extent of pollutant removal. The non-chemical technologies showed advantages in terms of complete (in the case of adsorption) or easy (enzymatic treatments) removal of toxic treatment by-products. The AOP methods led to the production of different photoproducts depending on the chosen treatment. Toxic products remained in most cases following treatment, though the toxicity level is significantly reduced with combination treatments. Among the treatments, a photochemical method combining UV, produced with a KrCl excilamp, and hydrogen peroxide achieved total removal of chlorophenol and all by-products and is considered the best treatment for chlorophenol removal.

Efficiency of KrCl excilamp (222 nm) for inactivation of bacteria in suspension.

AIMS: To examine the killing efficiency of UV KrCl excilamp against gram-positive and gram-negative bacteria. METHODS AND RESULTS: Vegetative cells of Bacillus cereus, Bacillus subtilis, Escherichia coli O157:H7, Staphylococcus aureus and Streptococcus pyogenes at initial populations from 10(2) to 10(7) colony-forming units (CFU) ml(-1) were treated by KrCl excilamp in sterile Ringer's solution with and without H(2)O(2). The number of viable cells was determined using spread plating techniques and nutrient agar method with subsequent incubation at 28 degrees C or 37 degrees C for 24 h. At estimated populations of 10(2)-10(5) CFU ml(-1) E. coli O157:H7 and Staph. aureus were the most sensitive and showed 100% disinfection within 15 s (29.2 mJ cm(-2)). Bacillus subtilis was more sensitive to UV treatment than B. cereus. The UV/H(2)O(2) inactivation rate coefficients within this population range were two times higher than those observed for UV treatment alone. No effect of H(2)O(2) was observed at 10(7) CFU ml(-1) for Bacillus sp. and Strep. pyogenes. CONCLUSIONS: The narrow-band UV radiation at 222 nm was effective in the rapid disinfection of bacteria in aqueous suspensions. SIGNIFICANCE AND IMPACT OF THE STUDY: KrCl excilamps represent UV sources which can be applied for disinfection of drinking water in advanced oxidation processes.

UV treatment of microorganisms on artificially-contaminated surfaces using excimer and microwave UV lamps.

An XeBr excilamp having a peak emission at 283 nm, and microwave UV lamps with peak emissions at 253.7 nm that also generate ozone, have been tested for ability to eradicate high populations of microbial vegetative cells and spores (of bacteria and fungi) artificially added to filter surfaces. The study examined the energy required to completely eradicate large populations on filter surfaces. It was found that both the excilamp and microwave UV lamps were effective at killing large populations on surfaces with killing efficiency dependant on the type of organism, and, whether present in its vegetative or spore forms. The main killing factor is UV radiation following short treatment times. It is considered that for longer irradiation periods that are required to facilitate complete destruction of surface microorganisms, ozone and other oxidising species produced by microwave UV lamps would act to enhance microbial destruction.

Canine leishmaniosis in Cyprus due to Leishmania infantum MON 1.

During a serological survey in 1996, a total of 601 dogs (group I) distributed all over the government controlled southern area of Cyprus was examined using an enzyme-linked immunosorbent assay (ELISA) for the presence of specific antibodies directed against soluble antigens of promastigote stages of Leishmania infantum. The overall seroprevalence rate in this group was 1.7%. A second group (group II) of dogs was selected from regions where seropositive dogs where determined within the first group. In group II specific anti-Leishmania antibodies could be detected in 30 of 301 dogs investigated (10%). The highest seroprevalence rates were found in the regions of Agios Georgios (26.2%) and Limnatis (12.2%). Ten parasite isolates from ten dogs (six with typical symptoms of canine leishmaniosis and four without symptoms) originating from five locations could be characterised by zymodeme analysis. All ten isolates were identified as L. infantum zymodeme MON 1.

Recovery of Rhodococcus biosurfactants using methyl tertiary-butyl ether extraction.

In the present study, we proposed methyl tertiary-butyl ether (MTBE) as a solvent for extraction of biosurfactants from Rhodococcus bacterial cultures. After comparison with other well known solvent systems used for biosurfactant extraction, it was found that MTBE was able to extract crude surfactant material with high product recovery (10 g/l), efficiency (critical micelle concentration (CMC), 130-170 mg/l) and good functional surfactant characteristics (surface and interfacial tensions, 29 and 0.9 mN/m, respectively). The isolated surfactant complex contained 10% polar lipids, mostly glycolipids possessing maximal surface activity. Ultrasonic treatment of the extraction mixture increased the proportion of polar lipids in crude extract, resulting in increasing surfactant efficiency. Due to certain characteristics of MTBE, such as relatively low toxicity, biodegradability, ease of downstream recovery, low flammability and explosion safety, the use of this solvent as an extraction agent in industrial scale biosurfactant production is feasible.

The effectiveness of a multi-spark electric discharge system in the destruction of microorganisms in domestic and industrial wastewaters.

The aim of this work was to investigate the effectiveness of a high voltage multi-spark electric discharge, with pulse energy of 1 Joule, in killing microorganisms in wastewater. Wastewater from primary treated effluent arising from domestic and industrial sources was abstracted for continuous pulsed discharge disinfection. The wastewater contained a large mixed population of microorganisms (approximately 10(7) CFU ml(-1) [10(9) CFU 100 ml(-1)] total aerobic heterotrophic bacteria) including vegetative cells and spores. The electrical conductivity of the wastewater ranged from 900-1400 microS cm(-1) and it was shown that a specific energy of 1.25-1.5 J cm(-3) was required to achieve 1 log reduction in bacterial (faecal coliforms/total aerobic heterotrophs) content. This is higher than that previously shown to reduce the population of E. coli in tap water of low conductivity, demonstrating the role of total wastewater constituents, including dissolved and particulate substances, water colour and the presence of microbial spores, in effective disinfection. The system can be engineered to eradicate microbial populations to levels governed by legislation by increasing treatment time or energy input.

Screening of dogs for Echinococcus granulosus coproantigen in a low endemic situation in Cyprus.

In the framework of an echinococcosis surveillance and control programme in Cyprus, a commercial enzyme-linked immunosorbent assay (ELISA) (CHEKIT ECHINOTEST) designed for the detection of Echinococcus granulosus and E. multilocularis coproantigens was used in 1997-2000 for the investigation of large numbers of dogs. Most of the animals originated from areas where approximately 0.2% of the dogs had been found to be infected with E. granulosus in previous (1993-1996) arecoline surveys. The sensitivity of the coproantigen test was 83%, as determined in 35 dogs naturally infected with this cestode species. The specificity was 98% in 97 randomly selected dogs from Cyprus, but it was reduced to 80% in a group of 50 dogs, infected with Taenia spp. A total of 6551 dogs (mainly of rural origin) was examined, including three large groups (N: 2928, 1761 and 1800) from the Government Control Area (GCA) in southern Cyprus and a small group (N: 62) from the Non-Government Control Area (NonGCA) in the northern part of the island. Among the dogs from the GCA, 184 (2.8%) tested positive for coproantigen; coproantigen prevalences were 2.6, 4.9 and 1.1% in these three groups, and of 62 dogs from the NonGCA 8.1% were positive. The calculated true prevalences of E. granulosus in the dog population of the GCA ranged between 0.0 and 3.58%. The predictive values of the test, based on a 0.2% prevalence, was >99.9% for negative results, but very low (7%) for coproantigen-positive results. However, the relatively small number of coproantigen-positive dogs can be treated with praziquantel or the results can further be confirmed by arecoline purging.

A comparison of five rapid direct toxicity assessment methods to determine toxicity of pollutants to activated sludge.

Five rapid direct toxicity assessment methods were used in three European partner countries to determine the toxicity of single toxicants, mixed toxicants and real industrial wastes. The final aim was to protect microbial degradation of organic wastes in biological treatment processes and hence enhance the quality of treated effluents to be discharged to the environment. Nitrification inhibition, Respirometry, Adenosine triphosphate luminescence and Enzyme inhibition were tested utilising activated sludge as the testing matrix. The Vibrio fischeri toxicity test was used as a surrogate to compare the various microbial bioassays. The IC50 (toxicant concentration eliciting a 50% inhibitory effect) was determined for a number of pollutants including single toxicants Cd, Cr, Cu, Zn, 3,5-dichlorophenol, toluene and linear alkylbenzenesulphonate (LAS); a standard mixture of metals and LAS; a standard mixture of organics and LAS, and 16 industrial effluents. The V. fischeri bioassay was also chosen in order to assess quality control of toxicant preparation during testing in the different laboratories of the partner countries. Comparisons of sensitivity, cost of implementation, cost per test, relevance, and ease of use were made. The most sensitive bioassays were V. fischeri and Nitrification inhibition, however, this depended in the main on the pollutant and mixtures tested. It is recommended that during assessment of wastewater toxicity a suite of tests be used rather than reliance on one particular test.

Silver nanotoxicity using a light-emitting biosensor Pseudomonas putida isolated from a wastewater treatment plant.

The effect of silver ions, nano- and micro-particles on a luminescent biosensor bacterium Pseudomonas putida originally isolated from activated sludge was assessed. The bacterium carrying a stable chromosomal copy of the lux operon (luxCDABE) was able to detect toxicity of ionic and particulate silver over short term incubations ranging from 30 to 240 min. The IC(50) values obtained at different time intervals showed that highest toxicity (lowest IC(50)) was obtained after 90 min incubation for all toxicants and this is considered the optimum incubation for testing. The data show that ionic silver is the most toxic followed by nanosilver particles with microsilver particles being least toxic. Release of nanomaterials is likely to have an effect on the activated sludge process as indicated by the study using a common sludge bacterium involved in biodegradation of organic wastes.

Comparison of alternative treatments for 4-chlorophenol removal from aqueous solutions: use of free and immobilized soybean peroxidase and KrCl excilamp.

The removal of 4-chlorophenol (4-CP) from industrial wastewater continues to be an important environmental issue and some interesting results have been obtained using oxidoreductive enzymes such as peroxidases and UV, generated by novel excilamps. In this study enzyme (free and immobilized soybean peroxidase) and UV (produced by a KrCl excilamp) were used to treat 4-CP solutions at concentrations ranging from 50 to 500 mg L(-1). It was shown that the excilamp can facilitate higher removal efficiencies in all cases with complete 4-CP elimination taking place between 5 and 90 min. The enzyme removed approximately 80% of the 4-CP concentrations in both the free and immobilized state up to concentrations of 250 mg L(-1). At 500 mg L(-1) the immobilized system shows much higher removal efficiency due to increased enzyme stability in the presence of higher formation of by-products.

Testing the toxicity of influents to activated sludge plants with the Vibrio fischeri bioassay utilising a sludge matrix.

To protect the bioceonosis within activated sludge, a method of predicting the toxic effect of influents to the biological treatment stage of waste water treatment plants, based on DIN method 38412 L 34, has been developed. A population of the luminescent marine bacterium Vibrio fischeri was incorporated into a sludge testing matrix derived from a model laboratory and real activated sludge plants. The sludge was challenged with different concentrations of pure toxicants and complex aqueous samples, and light output by V. fischeri monitored. The results were compared to toxicant testing in the absence of sludge (standard test). The modified method was found to be less sensitive for some toxicants tested than the standard DIN and other bioluminescent tests, but considered more realistic as it provides buffering and takes into account sorption which can affect the sensitivity of the test towards some compounds. The method is comparable in terms of ease of use, speed, reproducibility and cost effectiveness to standard V. fischeri luminescence methods.

Combination ecotoxicity and testing of common chemical discharges to sewer using the Vibrio fischeri luminescence bioassay.

In order to investigate possible synergistic or antagonistic (more or less than additive) toxicity effects, mixtures of chemicals were tested in water using a microbial bioassay. Ten toxicants (3,4-dichloroaniline, 3,5-dichlorophenol, cadmium, chromium, copper, Lindane, linear alkylbenzene sulphonate, pentachlorophenol, toluene, zinc) were chosen on the basis of their common occurrence in industrial effluents within local waste water treatment plants. These toxicants also cover a wide range of modes of toxic action, namely, polar and non-polar narcosis, membrane disruption, respiratory disruption, uncouplers of oxidative phosphorylation, biochemical disruption and enzyme inhibition. Efficient screening for possible combination toxicity between toxicants involved testing the chemicals both singly and in triplet combinations. The triplets were based on four replicates of a balanced incomplete block design (BIB). A standardised Vibrio fischeri rapid toxicity bioluminescence assay was used. The combinations tested showed that only one mixture was found to be significantly more toxic than expected from the pure single-toxicant results. Two triplets were significantly less toxic. Further tests on the more toxic triplet showed that the effect was due to only one of the 45 pairs originally screened. It is concluded that synergistic effects in combinations of toxicants are rather rare in bioluminescence systems utilising common effluents discharged to sewer.

The effects of fimbriate and non-fimbriate strains of Escherichia coli on the oxidative reactions of neutrophils from a variety of species.

5 fimbriate strains and 5 non-fimbriate strains of Escherichia coli were tested for their ability to enhance the cyanide insensitive oxygen consumption of human, bovine and ovine blood neutrophils in the absence of serum opsonins. Neutrophils from all 3 animal species showed significantly increased oxygen consumption when challenged with fimbriate strains. Only ovine neutrophils were stimulated by non-fimbriate strains although to a lesser extent than by fimbriate strains. In the presence of serum the stimulation of ovine neutrophils was the same whether fimbriate or non-fimbriate strains were used.

Identification of Rhodococcus equi using the polymerase chain reaction.

Two regions in the gene coding for 16S rRNA in Rhodococcus equi were selected as species-specific primer sequences for the polymerase chain reaction (PCR). PCR using these primers was tested against 10 strains of R. equi (including the type strain) and gave positive results for all but was negative for all other tested species of Rhodococcus; representatives of the most closely related genera and a number of other bacterial species. This method could therefore be used to identify this species which can infect the lungs or other organs of horses, pigs, humans and other animals.

Alkanotrophic Rhodococcus ruber as a biosurfactant producer.

In this report we examined the structure and properties of surface-active lipids of Rhodococcus ruber. Most historical interest has been in the glycolipids of Rhodococcus erythropolis, which have been extensively characterised. R. erythropolis has been of interest due to its great metabolic diversity. Only recently has the metabolic potential of R. ruber begun to be explored. One major difference in the two species is that most R. ruber strains are able to oxidise the gaseous alkanes propane and butane. In preparation for investigation of the effects of gas metabolism on biosurfactant production, we set out to characterise the biosurfactants produced during growth on liquid n-alkanes and to compare these with R. erythropolis glycolipids.

Pulsed high voltage electric discharge disinfection of microbially contaminated liquids.

AIMS: To examine the use of a novel multielectrode slipping surface discharge (SSD) treatment system, capable of pulsed plasma discharge directly in water, in killing micro-organisms. METHODS AND RESULTS: Potable water containing Escherichia coli and somatic coliphages was treated with pulsed electric discharges generated by the SSD. The SSD system was highly efficient in the microbial disinfection of water with a low energy utilization (eta approximately 10-4 kW h l-1). CONCLUSIONS: The SSD treatment was effective in the destruction of E. coli and its coliphages through the generation of u.v. radiation, ozone and free radicals. SIGNIFICANCE AND IMPACT OF THE STUDY: The non-thermal treatment method can be used for the eradication of micro-organisms in a range of contaminated liquids, including milk, negating the use of pasteurization. The method utilizes multipoint electric discharges capable of treating large volumes of liquid under static and flowing regimes.

Identification and environmental detection of Rhodococcus species by 16S rDNA-targeted PCR.

Bacteria of the genus Rhodococcus can degrade a wide range of organic pollutants and catalyse many useful biotransformations. There is a need for improved tests to identify Rhodococcus species. PCR-based methods for species identification offer advantages in terms of speed and accuracy over traditional methods and can allow direct detection of microbes in environmental samples., PCR tests, using primers targeted at species-specific sequences in the 16S rRNA gene, were successfully developed for R. globerulus, R. erythropolis, R. opacus and R. ruber. These tests gave positive results with all or most strains of target species but did not generally cross-react with other species. Cases of apparent cross-reaction were shown to be due to prior misclassification of strains of R. opacus as R. erythropolis and of strains of R. ruber as R. rhodochrous. A simple and rapid method for the extraction and purification of DNA from soil was developed and successfully applied to the PCR detection of indigenous R. erythropolis in an environmental sample. Cell lysis in the samples was achieved by lysozyme and sarkosyl treatment, aided by freeze-thaw cycles. Removal of humic compounds inhibitory to PCR was accomplished by CTAB treatment with solvent extraction and, if necessary, passage of extracts through Sepharose CL-6B in a spun-column format. Extracts prepared using a tris-EDTA buffer were much clearer than those prepared using a sodium phosphate buffer, indicating lower levels of humic compounds. A detection limit of 104 cfu g-1 of soil was achieved and the use of a secondary PCR allowed detection of 1 cfu g-1.