Tomislav Ivanković

Surfactants in the Environment

Surfactants in the Environment Surfactants are a diverse group of chemicals that are best known for their wide use in detergents and other cleaning products. After use, residual surfactants are discharged into sewage systems or directly into surface waters, and most of them end up dispersed in different environmental compartments such as soil, water or sediment. The toxic effects of surfactants on various aquatic organisms are well known. In general, surfactants are present in the environment at levels below toxicity and in Croatia below the national limit. Most surfactants are readily biodegradable and their amount is greatly reduced with secondary treatment in wastewater treatment plants. The highest concern is the release of untreated wastewater or wastewater that has undergone primary treatment alone. The discharge of wastewater polluted with massive quantities of surfactants could have serious effects on the ecosystem. Future studies of surfactant toxicities and biodegradation are necessary to withdraw highly toxic and non-biodegradable compounds from commercial use and replace them with more environmentally friendly ones. Surfaktanti u Okolišu Surfaktanti ili površinski aktivne tvari raznolika su skupina molekula najpoznatijih po uporabi u sastavu deterdženata i ostalih sredstava za pranje i čišćenje. Nakon uporabe u kućanstvu ili industriji, surfaktanti se ispuštaju u kanalizacijski sustav ili izravno u površinske vode te većina surfaktanata završi raspršena u vodi, sedimentu ili tlu. Toksični utjecaj surfaktanata na vodne organizme dobro je istražen i opisan u literaturi. U većini slučajeva surfaktanti su u okolišu prisutni u koncentracijama nižim od toksične te nižim od maksimalne koncentracije dopuštene hrvatskim zakonskim odredbama. Većina surfaktanata klasificirana je kao biološki razgradiva i njihova se koncentracija znatno smanjuje biološkom obradom otpadne vode pa je najveći rizik za okoliš ispuštanje prethodno pročišćene ili nepročišćene otpadne vode. Takva otpadna voda opterećena visokim koncentracijama surfaktanata može nepovoljno utjecati na okoliš. Potrebno je proučavati toksičnost i biološku razgradnju surfaktanata u svrhu uklanjanja visoko štetnih i biološki nerazgradljivih surfaktanata iz komercijalne uporabe te njihovu zamjenu tvarima manje štetnim za okoliš.

Antibacterial activity of heavy metal-loaded natural zeolite

The antibacterial activity of natural zeolitized tuffs containing 2.60wt.% Cu(2+), 1.47 Zn(2+) or 0.52 Ni(2+) were tested. Antibacterial activities of the zeolites against Escherichia coli and Staphylococcus aureus were tested after 1h and 24h of exposure to 1g of the zeolite in 100mL of three different media, namely Luria Bertani, synthetic wastewater and secondary effluent wastewater. The antibacterial activities of the zeolites in Luria Bertani medium were significantly lower than those in the other media and negatively correlated with the chemical oxygen demand of the media. The Ni-loaded zeolite showed high leaching of Ni(2+) (3.44-9.13wt.% of the Ni(2+) loaded) and weak antibacterial activity in the effluent water. Since Cu-loaded zeolite did not leach Cu(2+) and the leaching of Zn(2+) from Zn-loaded zeolite was low (1.07-1.61wt.% of the Zn(2+) loaded), the strong antibacterial activity classified the Cu- and Zn-loaded zeolite as promising antibacterial materials for disinfection of secondary effluent water.

Toxicity of anionic and cationic surfactant to Acinetobacter junii in pure culture

The harmful effects of surfactants to the environment are well known. We were interested in investigating their potential toxicity in a pure culture of Acinetobacter junii, a phosphate (P)-accumulating bacterium. Results showed a high acute toxicity of sodium dodecyl sulfate (SDS) and hexadecyltrimethylammonium bromide (HDTMA) against A. junii. The estimated EC50 values of the HDTMA for the inhibition of CFUs in the pure culture of A. junii was 3.27 ± 1.12 × 10−7 mol L−1 and for the inhibition of the P-uptake rates 2.47 ± 0.51 × 10−6 mol L−1. For SDS, estimated EC50 values for the inhibition of CFUs in the pure culture of A. junii was 5.00 ± 2.95 × 10−6 mol L−1 and for the inhibition of the P-uptake rates 3.33 ± 0.96 × 10−4 mol L−1. The obtained EC50 values in the standardised yeast toxicity test using Saccharomyces cerevisiae were 3.03 ± 0.38 × 10−4 and 4.33 ± 0.32 × 10−5 mol L−1 for SDS and HDTMA, respectively. These results emphasized the need to control concentrations of surfactants entering the activated sludge system. The negative effects of these toxicants could greatly decrease populations of P-accumulating bacteria, as well as eukaryotic organisms, inhabiting activated sludge systems, which in turn could result in the decrease of the system efficiency.

The effect of mineral carrier composition on phosphate-accumulating bacteria immobilization.

The goal of this study was to determine the dynamics and yield of immobilization of the phosphate-accumulating bacterium Acinetobacter junii on mineral carriers. As mineral carriers natural clinoptilolite tuff from Turkey (T) and Serbia (S) and natural bentonite (TER), in original and magnesium (Mg)-exchanged form were used. The key feature which determined the extent of immobilization of A. junii was the type of carrier; the immobilization yield decreased in order T>TER>S. The number of immobilized cells was significantly higher for the Mg-exchanged carriers when compared to their original counterparts (95 and 75 x 10(8)CFU g(-1) for T, 74 and 58 x 10(8)CFU g(-1) for TER, 19 and 6 x 10(8)CFU g(-1) for S). The Mg-exchanged T and S displayed a prolonged biofilm growth up to 24h, while the original counterparts reached the mature biofilm after 12h of incubation. Both forms of TER reached the mature biofilm after 24h of incubation, due to swelling property of the material. The number of immobilized cells correlated significantly negatively with particle size of the carrier, indicating that particle size is another important feature which determined the extent of immobilization. The Mg-exchange of original carriers resulted in significant increase of the zeta potential. When all of the materials were compared, the increase of the zeta potential of carriers correlated negatively with the number of immobilized cells, suggesting that the zeta potential of material is not a crucial factor which determined the immobilization of cells.

Toxicity of dodecylpyridinium and cetylpyridinium clorides against phosphate-accumulating bacterium

The antibacterial effect of cationic surfactants against the pure culture of phosphate (P)-accumulating bacterium Acinetobacter junii was investigated. The estimated EC50 values of the N-dodecylpyridinium chloride (DPC) for growth inhibition was 1.4±0.5 × 10−6 mol L−1 and for the inhibition of the P-uptake rates 7.3±2.6 × 10−5 mol L−1. The estimated EC50 values of the N-cetylpyridinium chloride (CPC) for growth inhibition was 4.9±1.3 × 10−7 mol L−1 and for the inhibition of the P-uptake rates 7.7±2.9 × 10−6 mol L−1. This suggests the importance of controlling the amounts of cationic surfactants in influent of the wastewater treatment systems in order to avoid the possible failure of the biological P removal from wastewaters.

Selective immobilization of Acinetobacter junii on the natural zeolitized tuff in municipal wastewater

The immobilization of desired bacteria onto material was usually performed in synthetic media. The aim of this study was to test the immobilization of phosphate (P)-accumulating bacteria Acinetobacter junii onto natural zeolitized tuff (NZ) in the raw or sterilized municipal wastewater containing the common bacteria Escherichia coli and Enterococcus faecalis and the performance of immobilized A. junii in the same type of wastewater. In the sterilized wastewater which contained the mixture of A. junii, E. coli and E. faecalis, the A. junii was selectively immobilized onto NZ in significantly higher numbers than E. coli and E. faecalis. The A. junii added in the form of bioparticles to the wastewater containing E. coli and E. faecalis, multiplied and removed P from wastewater. The P removal from wastewater was a function of biomass of P-accumulating bacteria and not the amount of NZ or bioparticles used. The performance of A. junii was significantly better in membrane filtered than in autoclaved wastewater. The experiments that were performed in raw non sterilized wastewater showed that A. junii can be successfully immobilized onto NZ in competition with natively present heterotrophic bacteria, retain its metabolic activity and successfully remove P from such water, which makes this technology feasible from biotechnological aspect.

Requirement of Acinetobacter junii for magnesium, calcium and potassium ions

The goal of this study was to determine the concentrations of Mg, Ca and K ions required for the formation of metabolically active population of phosphate (P)-accumulating bacterium Acinetobacter junii. The availability of Mg, Ca and K originating from natural minerals in the conditions of severe shortage of these cations was tested. In the case of shortage of Mg, Ca and K ions in wastewater the P removal was absent due to the decay of A. junii. In the cases of Mg or K shortage in wastewater the P removal was negligible due to the decay of A. junii, while Ca was not essential for the examined bacterium. The minimal required concentrations of Mg and K in synthetic wastewater were 0.64 mg Mg/mg P and 0.50 mg K/mg P. The natural zeolitized tuffs and bentonite, either in Mg, Ca or K form, successfully replaced the lack of Mg, Ca, K and trace metals in wastewater. The requirement of A. junii for examined cations was in the order: Mg>K>Ca.

The fate of carbapenem-resistant bacteria in a wastewater treatment plant

Wastewater treatment plants have been considered potential sources of antibiotic resistance gene exchange and release into the environment. The aim of our study was to quantify environmental and human-associated carbapenem-resistant bacterial populations (CRBPs) across wastewater treatment stages and correlate bacterial counts to physicochemical and other bacteriological parameters in order to see their behaviour in wastewater and sludge and their potential dissemination in the environment. Samples were taken from five sites (treatment stages) of the largest Croatian wastewater treatment plant (20 per site) over 10 months of monitoring. CRBPs were found at all wastewater treatment stages save for the lime-treated, stabilised sludge, which underlines the importance of effluent and digested sludge disinfection. Secondary sludge settling removed 99% of CRBP from the effluent, but the relative proportion of CRBP in the total bacterial count significantly increased in the effluent (0.0020%) and digested sludge (0.0019%) compared to the influent (0.0006%), indicating selection for resistant bacteria in these settings. CRBP counts did not correlate with measured carbapenem concentrations in wastewater, which suggests that antibiotic concentrations were not the reason for CRBP selection. Negative correlation between activated sludge retention time and CRBP indicated that their number could be reduced by increasing the retention time during secondary treatment. Despite the indications that WWTPs select for antibiotic-resistant bacteria, wastewater treatment is very efficient in reducing their absolute numbers, and proper effluent and sludge disinfection can significantly reduce dissemination of antibiotic-resistant bacteria into the environment.

Colonization of diatoms and bacteria on artificial substrates in the northeastern coastal Adriatic Sea

Abstract Every surface that is immersed in seawater becomes rapidly covered with an unavoidable biofilm. Such biofilm formation, also known as fouling, is a complex multistage process and not yet thoroughly investigated. In this study, the succession of diatoms and bacteria was investigated during a one month exposure on an artificial substrate of plexiglass (polymer of methyl methacrylate) mounted above the seafloor at a depth of 5 m. For biofilm analyses, the fouling was investigated using selective agar plates, epifluorescence, light and electronic microscopy, as well as high performance liquid chromatography (HPLC) pigment analysis. During biofilm development, the abundance of all biofilm components increased and reached maximum values after a one month exposure. In the bacterial community, heterotrophic marine bacteria were dominant and reached 1.96 ± 0.79 × 104 colony forming units (CFU) cm-2. Despite the fact that faecal coliforms and intestinal enterococci were detected in the water column, faecal coliforms were not detected in the biofilm and intestinal enterococci appeared after one month of exposure but in the negligible number of 60 ± 10 CFU cm-2. The phototrophic component of the biofilm was dominated by diatoms and reached a concentration of 6.10 × 105 cells cm-2, which was supported by pigment analysis with fucoxanthin as dominant pigment in a concentration up to 110 ng cm-2. The diatom community was dominated by Cylindrotheca closterium and other pennate benthic diatoms. A detailed taxonomic analysis by electronic microscopy revealed 30 different taxa of diatoms. The study confirmed that a plexiglass surface in a marine environment is susceptible to biofouling within 30 days of contact. Furthermore, the co lonization process sequence firstly involved bacteria and cyanobacteria, and secondly diatoms, which together formed a primary biofilm in the sea.

Alkaline disinfection of urban wastewater and landfill leachate by wood fly ash

Abstract Wood fly ash is an industrial by-product of the combustion of different wood materials and is mostly disposed of as waste on landfills. In our preliminary experiments, wood ash exhibited antibacterial activity against urban wastewater bacteria and we focused on wood fly ash as a potential substrate for wastewater disinfection. The addition of ash at a concentration of 10 g L-1 (1 %) caused an instant increase of pH in urban wastewater and landfill leachate. High pH (10.1-12.7) inactivated bacterial populations in the wastewater and the removal of faecal coliforms and intestinal enterococci after 6 h of contact was 100 % (below the detection limit; <1 CFU per mL) with the most efficient ash sample (ash from combustion of beech) both in urban wastewater and landfill leachate. Properly chosen wood fly ash, i.e. one that tends to increase the pH to the greatest extent, proved to be a very effective disinfection substrate. Considering that water treated with wood ash has a high pH and needs to be neutralised before discharge, ash would be suitable for disinfection of leachates when smaller volumes are treated Sažetak Leteći pepeo iz drvne industrije je nusproizvod koji nastaje spaljivanjem različitog drvnog materijala i većinom završi na odlagalištima otpada. Budući da je u preliminarnim ispitivanjima leteći pepeo pokazao antibakterijsko djelovanje prema bakterijama iz otpadne vode, pokusi su usmjereni istraživanju pepela kao potencijalnog supstrata za dezinfekciju otpadne vode. Dodatak pepela u koncentraciji od 10 g L-1 (1 %) uzrokovao je trenutno povećanje vrijednosti pH komunalne i procjedne otpadne vode s odlagališta otpada. Visoki pH (10,1-12,7) uništio je bakterije u otpadnoj vodi te je korištenjem najučinkovitijega pepela (dobivenog spaljivanjem bukve) postignuto 100-postotno uklanjanje (odnosno manje od mogućnosti detekcije; <1 CFU mL-1) fekalnih koliforma i crijevnih enterokoka iz komunalne i procjedne otpadne vode nakon šest sati kontakta. Odgovarajući pepeo, odnosno onaj koji uzrokuje najveće povećanje vrijednosti pH, pokazao se kao vrlo učinkovit supstrat za dezinfekciju. Uzimajući u obzir činjenicu da otpadna voda tretirana pepelom ima povišen pH, te da je vrijednost pH potrebno neutralizirati prije ispuštanja u prirodni prijemnik, leteći drvni pepeo bio bi pogodan za dezinfekciju procjedne otpadne vode zbog manjih volumena koji zahtijevaju obradu