Biomedical subjects

G Dalhammar

Publications and source records attributed to G Dalhammar.

At least 19 recordsLinked to original sources

PubMed2005

Single-step ion exchange purification of the coagulant protein from Moringa oleifera seed.

The coagulant protein from Moringa oleifera (MO) seed was purified using a single-step batch ion exchange (IEX) method. Adsorption and elution parameters were optimized. Impact of the purification on the reduction of organic and nutrient release to the water was studied. The matrix was equilibrated using ammonium acetate buffer, and the optimum ionic strength of NaCl for elution was 0.6 M. The time for adsorption equilibrium was between 90 and 120 min. Maximum adsorption capacity of the matrix, estimated with the Langmuir model, was 68 mg protein/g adsorbent. The purified protein does not release organic and nutrient loads to the water, which are the main concerns of the crude extract. This work suggests that a readily scalable single-step IEX purification method can be used to produce the coagulant protein and it can be carried out with locally available facilities. This will promote the use of MO in large water treatment plants and other industries.

Chromatography, Ion Exchange↗

PubMed2005

Removal of micro-organisms in a small-scale hydroponics wastewater treatment system.

AIMS: To measure the microbial removal capacity of a small-scale hydroponics wastewater treatment plant. METHODS AND RESULTS: Paired samples were taken from untreated, partly-treated and treated wastewater and analysed for faecal microbial indicators, i.e. coliforms, Escherichia coli, enterococci, Clostridium perfringens spores and somatic coliphages, by culture based methods. Escherichia coli was never detected in effluent water after >5.8-log removal. Enterococci, coliforms, spores and coliphages were removed by 4.5, 4.1, 2.3 and 2.5 log respectively. Most of the removal (60-87%) took place in the latter part of the system because of settling, normal inactivation (retention time 12.7 d) and sand filtration. Time-dependent log-linear removal was shown for spores (k = -0.17 log d(-1), r(2) = 0.99). CONCLUSIONS: Hydroponics wastewater treatment removed micro-organisms satisfactorily. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigations on the microbial removal capacity of hydroponics have only been performed for bacterial indicators. In this study it has been shown that virus and (oo)cyst process indicators were removed and that hydroponics can be an alternative to conventional wastewater treatment.

Clostridium perfringens↗

PubMed2004

Biological nitrogen and organic matter removal from tannery wastewater in pilot plant operations in Ethiopia.

The objective of this study was to set-up a pilot plant and to evaluate its effectiveness for biological nitrogen and organic matter removal from tannery wastewater in Ethiopia. A pilot wastewater treatment plant consisting of a predenitrification-nitrification process was constructed and operated for 6 months. This was fed with a raw tannery wastewater obtained from the Modjo Tannery located 70 km south of the capital, Addis Ababa. Up to 98% total nitrogen and chemical oxygen demand, and 95% ammonium nitrogen removal efficiencies were achieved in the system. The average effluent ammonium nitrogen ranged from 8.4 mg l(-1) to 86.0 mg l(-1), whereas the average effluent for nitrate nitrogen ranged from 2.9 mg l(-1) to 4.4 mg l(-1). The average values of denitrification and nitrification rates determined by nitrate and ammonium uptake rates (NUR and AUR) were 8.0 mg NO3-N [g volatile suspended solids (VSS)](-1) h(-1) and 5.4 mg NH4-N (g VSS)(-1) h(-1), respectively, demonstrating that the treatment processes of the pilot plant were effective. Further studies of the effect of chromium III on AUR showed 50% inhibition at a concentration of 85 mg l(-1), indicating that this metal was not causing process inhibition during performance operations. Thus, the predenitrification-nitrification process was found to be efficient for simultaneous removal of nitrogen and organic substrates from tannery wastewaters.

Chromium↗

PubMed2003

A small scale hydroponics wastewater treatment system under Swedish conditions.

A treatment plant using conventional biological treatment combined with hydroponics and microalgae is constructed in a greenhouse in the area of Stockholm, Sweden. The treatment plant is built for research purposes and presently treats 0.559 m3 of domestic wastewater from the surrounding area per day. The system uses anoxic pre-denitrification followed by aerobic tanks for nitrification and plant growth. A microalgal step further reduces phosphorus, and a final sand filter polishes the water. During a three week period in July 2002 the treatment capacity of this system was evaluated with respect to removal of organic matter, phosphorus and nitrogen. 90% COD removal was obtained early in the system. Nitrification and denitrification was well established with total nitrogen reduction of 72%. Phosphorus was removed by 47% in the process. However, higher phosphorus removal values are expected as the microalgal step will be further developed. The results show that acceptable treatment can be achieved using this kind of system. Further optimisation of the system will lead to clean water as well as valuable plants to be harvested from the nutrient rich wastewater.

Facility Design and Construction↗

PubMed2003

Performance of a small wastewater stabilisation pond system in tropical climate in Dar es Salaam, Tanzania.

Waste stabilisation ponds (WSP) are widely used in Tanzania. Their traditional design focuses on BOD and pathogen removal, but nutrient removal is equally important. WSP performance was evaluated to come up with information that would be used to evaluate pond performance. Samples were analysed twice a month for six months. Results showed total and faecal coliform removal by 4 log units (99.96 and 99.98% respectively). There was partial reduction of COD, BOD, (46% and 27% respectively), conductivity and total dissolved solids (32.6 and 32.4% respectively). Variation for the nutrients like inorganic phosphorus, ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen was not consistent and there was practically no reduction, although there is some nitrification taking place. From this study it may be concluded that BOD and nutrients may not be useful to evaluate pond performance. Instead parameters such as conductivity, total dissolved solids, coliform bacteria, dissolved oxygen, pH and chlorophyll would be suitable for that purpose, due to their consistent variation within cells of the pond system. But BOD and nutrient removal are important and have to be improved to enhance treatment in the WSP.

Chlorophyll↗

PubMed2001

Development of nitrification inhibition assays using pure cultures of Nitrosomonas and Nitrobacter.

Restricted requirements for nitrogen reduction at wastewater treatment plants have increased the need for assays determining the inhibition of nitrification. In this paper, two new assays studying ammonia oxidation and nitrite oxidation, respectively, are presented. As test organisms, pure cultures of Nitrosomonas and Nitrobacter isolated from activated sludge are used. The assays are performed in test tubes where the bacteria are incubated with the compound or wastewater to be tested. The nitrification rate is measured during 4 h and compared with reference samples. The test organisms were characterised with respect to temperature, pH and cell activity. Optimum temperature was 35 degrees C for Nitrosomonas and 38 degrees C for Nitrobacter; optimum pH was 8.1 for Nitrosomonas and 7.9 for Nitrobacter. There was a linear relationship between the nitrification rate and the cell concentration in the studied interval. The cell activity decreased slightly with storage time. A significant level of inhibition was calculated to 11% for the Nitrosomonas assay, and to 9% for the Nitrobacter assay. The assays are applicable to determination of nitrification inhibition in samples of industrial waste waters or influents of treatment plants, or chemical substances likely to be found in wastewater.

Ammonia↗

PubMed2001

Determination of hydrocarbons in old creosote contaminated soil using headspace solid phase microextraction and GC-MS.

Headspace solid phase microextraction (HS-SPME) has been used together with GC-MS to analyze organic substances directly in a soil, heavily contaminated with PAHs/creosote (approximately 300 mg/kg soil), from an old gaswork site in Stockholm, Sweden. The HS-SPME results, both qualitative and quantitative, were compared with traditional liquid extraction using ethyl acetate/hexane (20:80). It was shown that the concentrations determined with HS-SPME at 60 degrees C correlated well, for compounds containing up to two and three aromatic rings (naphthalenes, acenaphthene, acenaphthylene and fluorenes, while a lower concentration was obtained for phenanthrene, anthracene, fluoranthene and pyrene. The total concentrations for each compound determined with HS-SPME ranged from 2 to 25 microg/g soil. Quantification was done using standard addition of compounds directly to the soil samples. The bioavailable fraction of the compounds in the contaminated soil at 20 degrees C was analyzed using external calibration by spiking sterile uncontaminated sand (same texture and particle size as the contaminated soil but without a heavily sorbed organic fraction) with hydrocarbon standards in different concentrations. Storage of exposed fibers at 20 degrees C showed that analysis should be done within two days to make qualitative measurements and earlier (as soon as possible) for quantitative determinations.

Biological Availability↗

PubMed2001

Comamonas denitrificans sp. nov., an efficient denitrifying bacterium isolated from activated sludge.

To find a biomarker for denitrification in activated sludge, five denitrifying strains isolated from three wastewater treatment plants were studied. These strains were selected from among 1,500 isolates for their excellent denitrifying properties. They denitrify quickly and have no lag phase when switching from aerobic to anoxic conditions. All strains have the cd1-type of nitrite reductase. The strains are Gram-negative rods and they all grow as filamentous chains when cultivated in liquid solution. The strains differ in colony morphology when grown on nutrient agar. Almost full-length 16S rDNA sequences were determined and phylogenetic analysis revealed that these strains are positioned among members of the genus Comamonas in the beta-subclass of the Proteobacteria. Signature nucleotides and bootstrap percentages were also analysed to verify this position. Strains 110, 123T, 2.99g, 5.38g and P17 were < or = 96.7% similar to known strains, but > or = 99.7% similar to each other, as judged from their 16S rDNA sequences, and grouped tightly together in the phylogenetic tree. Sequence motifs in the 16S rRNA gene were also found, suggesting the monophyletic origin of these strains. Nevertheless, some strains differed from the others, for example strain 110 branches early from the other strains and 5.38g is phenotypically more inert. Therefore, it is proposed that strains 110, 123T, 2.99g and P17 are classified into a new species, Comamonas denitrificans sp. nov., while the taxonomic status of strain 5.38g will have to await the outcome of further studies. The type strain of Comamonas denitrificans is 123T (ATCC 700936T).

Anti-Bacterial Agents↗

PubMed2000

Application of an amperometric immunosensor for the enumeration of Nitrobacter in activated sludge.

A competitive immunosensor using a monoclonal antibody has been developed for the enumeration of Nitrobacter in activated sludge and other environmental samples. Its cross-reactivity was tested against a number of bacterial strains and isolates. All strains of the nitrite-oxidising genera Nitrobacter and Nitrococcus reacted strongly with the monoclonal antibody. The nitrite-oxidising Nitrospira moscoviensis, as well as the ammonia oxidising bacteria and the heterotrophic bacteria tested, did not show any affinity towards the antibody in the immunosensor. The numbers of Nitrobacter were analysed in sludge samples from several wastewater treatment plants in Sweden. Detectable amounts were found in all samples. This study shows the adequacy of using this immunosensor for the enumeration of Nitrobacter in natural environments.

Antibodies, Bacterial↗

PubMed2000

Biological degradation of selected hydrocarbons in an old PAH/creosote contaminated soil from a gas work site.

An old PAH/creosote contaminated soil (total approximately 300 microg PAH/g soil) from a former gas work site in Stockholm, Sweden, has been treated at 20 degrees C with the addition of various nutrients and inoculated with bacteria (isolated from the soil) to enhance the degradation of selected hydrocarbons. Microcosm studies showed that the soil consisted of two contaminant fractions: one available, easily degraded fraction and a strongly sorbed, recalcitrant one. The bioavailable fraction, monitored by headspace solid phase microextraction, contained aromatics with up to three rings, and these were degraded within 20 days down to non-detectable levels (ng PAH/g soil) by both the indigenous bacteria and the externally inoculated samples. The nutrient additives were: a minimal medium (Bushnell-Haas), nitrate, nitrite, potting soil (Anglamark, Sweden), sterile water and aeration with Bushnell-Haas medium. After 30 days treatment most of the sorbed fractions were still present in the soil. Stirring or mechanical mixing of the soil slurries had the greatest effect on degradation, indicating that the substances were too strongly sorbed for the microorganisms. When stirring the choice of nutrient seemed less important. For the non-stirred samples the addition of nitrate with the bacterial inoculum showed the best degradation, compared to the other non-stirred samples. At the end of the experiments, accumulations of metabolites/degradation products, such as 9H-fluorenone, 4-hydroxy-9H-fluorenone, 9,10-phenanthrenedione and 4H-cyclopenta[def]phenanthrenone were detected. The metabolite 4-hydroxy-9H-fluorenone increased by several orders of magnitude during the biological treatments. Microbial activity in the soil was measured by oxygen consumption and carbon dioxide production.

Biodegradation, Environmental↗

PubMed1999

Aerobic degradation of a hydrocarbon mixture in natural uncontaminated potting soil by indigenous microorganisms at 20 degrees C and 6 degrees C.

A hydrocarbon mixture containing p-xylene, naphthalene, Br-naphthalene and straight aliphatic hydrocarbons (C14 to C17) was aerobically degraded without lag phase by a natural uncontaminated potting soil at 20 degrees C and 6 degrees C. Starting concentrations were approximately 46 ppm for the aromatic and 13 ppm for the aliphatic compounds. All aliphatic hydrocarbons were degraded within 5 days at 20 degrees C, to levels below detection (ppb levels) but only down to 10% of initial concentration at 6 degrees C. Naphthalene was degraded within 12 days at 20 degrees C and unaffected at 6 degrees C. At 20 degrees C p-xylene was degraded within 20 days, but no degradation occurred at 6 degrees C. Br-naphthalene was only removed down to 30% of initial concentration at 20 degrees C, with no significant effect at 6 degrees C. The biodegradation was monitored with head space solid-phase microextraction and gas chromatography-mass spectrometry.

Aerobiosis↗

PubMed1998

Biological degradation of diesel fuel in water and soil monitored with solid-phase micro-extraction and GC-MS.

Solid-phase micro-extraction (SPME) was used for monitoring degradation of hydrocarbons in diesel-fuel-contaminated (1% v/v) water and soil. Natural soil bacteria with and without external addition of inoculum were used. Directly after a 10-s exposure of the sample, the polydimethylsiloxane fibre was injected into the GC-MS. This method strongly reduced the time of analysis compared to a conventional liquid/liquid extraction. A comparison of SPME and pentane extraction of diesel oil was made and found to be consistent. The degradation of diesel fuel in water was monitored for 10 weeks using SPME. After 5 weeks all hydrocarbons were degraded except for the decahydronaphthalenes. These compounds were approximately 3% of the total hydrocarbons in the diesel oil used and remained undegraded throughout the study although none of the chemical or physical parameters was limiting. In the soil study the degradation of diesel fuel in normal soil was completed after 3 weeks, when the only remaining substances were decahydronaphthalenes. All samples were compared to sterile references to make up for evaporation losses. SPME proved to be a fast and reliable method to monitor changes in concentrations of semivolatile organic compounds.

Alkanes↗

PubMed1998

An amperometric enzyme-linked immunosensor for Nitrobacter.

A new amperometric enzyme-linked immunoassay for specific enumeration of Nitrobacter has been developed. This assay uses an electrode made of glassy carbon, on which the immunological reaction is carried out. The method is based on a competitive immunoassay principle, utilising monoclonal primary antibody and alkaline-phosphatase-labelled secondary antibody. The enzyme substrate 5-bromo-4-chloro-3-indolyl phosphate generates an electroactive product which is amperometrically detected. The effects of different parameters on the performance of the sensor have been studied. Quantitative detection of Nitrobacter using the immunosensor has been compared to a previously developed enzyme-linked immunosorbent assay showing compatible results. In addition, the overall assay time can be shortened with this new sensor. A detection limit of approximately 3 x 10(6) Nitrobacter cells/ml was obtained.

Antibodies, Monoclonal↗

PubMed1996

Field diagnosis of onchocerciasis in an area of high versus low endemicity: evaluation of the Dot Blot Assay.

Parasitological examination of skin snips is the most widely used diagnostic method for onchocerciasis, but it is associated with inconvenience and low sensitivity. We describe an inexpensive antibody-based dot blot assay (DBA) for the detection of Onchocerca volvulus infection. A field evaluation of this method was performed in the onchocerciasis endemic country Ghana by testing 370 individuals living in a highly onchocerciasis endemic area and 122 in an area of low endemicity. Sera from individuals with other filarial infections were also tested. The DBA was able to detect 95% of the parasitologically confirmed infected individuals in the highly endemic area. Cross-reactivity occurred with a minority of the sera from individuals infected with other filarial worms. The DBA was as good as or superior to presently available diagnostic tests, and it also fulfilled the criteria for a good screening method.

Adolescent↗

PubMed1990

Molecular characterization of immune inhibitor A, a secreted virulence protease from Bacillus thuringiensis.

The gene for the secreted neutral metalloprotease, immune inhibitor A (InA), from Bacillus thuringiensis var. alesti has been cloned and sequenced. The deduced amino acid sequence has been confirmed by partial amino acid sequencing. The central part of the amino acid sequence showed similarity to the active site in thermolysin. Southern and Western blots show that InA-related sequences are common among other B. thuringiensis subspecies. In Western blots, 17 out of 25 tested species gave a positive signal. Culture filtrates from subspecies expressing InA were toxic when injected in Trichoplusia ni larvae, whereas filtrate from a strain negative in Western blot had no effect when injected. The LD50 dose of purified InA protein injected in T. ni larvae was 12.5 +/- 2.5 ng per mg of larval body weight.

Amino Acid Sequence↗

PubMed1987

Transformation of vegetative cells of Bacillus thuringiensis by plasmid DNA.

Plasmid DNA-mediated transformation of vegetative cells of Bacillus thuringiensis was studied with the following two plasmids: pBC16 coding for tetracycline resistance and pC194 expressing chloramphenicol resistance. A key step was the induction of competence by treatment of the bacteria with 50 mM Tris hydrochloride buffer (pH 8.9) containing 30% sucrose. Transformation frequency was strongly influenced by culture density during the uptake of DNA and required the presence of polyethylene glycol. Growth in a minimal medium supplemented with Casamino Acids gave 35 times more transformants than growth in a rich medium. The highest frequencies were obtained with covalently closed circular DNA. With all parameters optimized, the frequency was 10(-3) transformants per viable cell or 10(4) transformants per microgram of DNA. Cells previously frozen were also used as recipients in transformation experiments; such cells gave frequencies similar to those obtained with freshly grown cells. The procedure was optimized for B. thuringiensis subsp. gelechiae, but B. thuringiensis subsp. kurstaki, B. thuringiensis subsp. galleriae, B. thuringiensis subsp. thuringiensis, and B. thuringiensis subsp. israelensis were also transformed. Compared with protoplast transformation, our method is much faster and 3 orders of magnitude more efficient per microgram of added DNA.

Bacillus thuringiensis↗

PubMed1984

Characterization of inhibitor A, a protease from Bacillus thuringiensis which degrades attacins and cecropins, two classes of antibacterial proteins in insects.

The insect pathogen Bacillus thuringiensis produces an exoprotease, inhibitor A, at the beginning of the stationary growth phase. In vitro, the enzyme selectively destroys cecropins and attacins, two antibacterial proteins found in immune hemolymph from Hyalophora cecropia. The specificity of this enzyme was investigated using cecropin A(1-33) and HPLC for separation and characterization of the fragments obtained. A maximum of 12 different peptides were produced and their positions in the known sequence of cecropin A(1-33) were deduced from their amino acid compositions. The enzyme did not show a stringent requirement for a specific amino acid sequence at the cleavage site but prefers a hydrophobic residue on the C-terminal side. The specificity of the enzyme is explained in terms of the open structure of the cecropins and a pronounced inability of inhibitor A to attack globular proteins.

Amino Acids↗

PubMed1979

Virulence factors in Bacillus thuringiensis: purification and properties of a protein inhibitor of immunity in insects.

We have previously shown that Bacillus thuringiensis subsp. alesti, serotype 3, produces two extracellular inhibitors of the immune system of Saturniid pupae (designated inhibitors A and B; Edlund et al., 1976). Starting from the culture supernatant of a new mutant of B. thuringiensis with a decreased extracellular proteolytic activity, we have now purified immune inhibitor A(InA). The procedure described consists of three steps: ultrafiltration, precipitation with ammonium sulphate and chromatography on hydroxylapatite. Purified InA gave a single band on polyacrylamide gel electrophoresis using either a gel concentration of 7.5% (w/v) and reducing and denaturing conditions or a gradient gel and native conditions. In both cases the apparent molecular weight was 78 000. A certain amount of proteolytic activity was always co-purified with InA but the two activities could be dissociated by heat or EDTA treatment. Antiserum against purified InA gave only one sharp precipitation band on immunodiffusion against InA with or without EDTA. InA inhibited the in vitro killing of Escherichia coli by immune haemolymph but did not affect the killing of Bacillus subtilis. InA was toxic for Drosophila when injected into the abdomen of adult male flies.

Animals↗