Detection of Listeria monocytogenes in cheese with the magnetic immuno-polymerase chain reaction assay.

A new detection system, the magnetic immuno-polymerase chain reaction (PCR) assay (MIPA) has been developed to detect Listeria monocytogenes in food. This method separates Listeria cells from PCR-inhibitory factors present in enrichment broths containing food samples by using magnetic beads coated with specific monoclonal antibodies (MAbs). The separated bacteria were lysed, and the supernatant containing the bacterial DNA was subjected to the PCR. Detection of L. monocytogenes in three naturally contaminated cheese samples with two different MAbs and PCR primers specific for the gene encoding the delayed-hypersensitivity factor showed that with MAb 55 all three samples were positive whereas with MAb A two samples were positive. A further improvement of the method was obtained by using a PCR step based on the listeriolysin O gene. A MIPA employing MAb 55 and the listeriolysin O gene primer set detected L. monocytogenes after 24 h of culture in Listeria Enrichment Broth samples from Port Salut artificially contaminated with 40 CFU/25 g. We could detect 1 CFU of L. monocytogenes per g of cheese after a second enrichment for 24 h in Fraser broth. The analysis time including both enrichments is approximately 55 h.     

The Reeuwijk lakes: A five years water quality study in an eutrophic ecosystem

The Reeuwijk Lakes (The Netherlands) present a typical example of eutrophication in the lower Rhine catchment area. In 1986 restoration of these lakes started by reducing the external P-loading. Two lakes, Lake Elfhoeven and Lake Nieuwenbroek, differing in P-load and residence time were selected for monitoring water quality parameters before (1983–1985) and after (1986–1987) these restoration measures. Reduction of the external P-loading did not result in lower P-concentrations in both lakes. In contrast, P and N increased. This may have been caused by an increase in diffuse discharges. However, seasonal cycles of P and N point to a strong internal loading of nutrients. The concentrations of chlorophyll a and carotene decreased, indicating a lower phytoplankton biomass. However, as C-phycocyanine concentrations increased the relative abundance of cyanobacteria became higher. Seston concentrations and zooplankton densities did not change. Transparency in the lakes slightly decreased after P-reduction and is far too low for the development of any vegetation of submerged waterplants. The typical differences between both lakes remained after restoration measures. The inverse relationship between the concentrations of chlorophyll a and total phosphorus at the two sampled stations remained constant. The differences in phytoplankton composition and the zooplankton biomass give a plausible explanation for this inverse relationship, between the two stations. Restoring the lakes after four decades of P-loading can presumably, not simply be done by lowering the external P-load alone. Supplementary in-lake measures may accelerate the restoration process.     

Characterization of Nitrosomonas europaea immobilized in calcium alginate

Nitrosomonas europaea cells have been immobilized in calcium alginate and the resulting preparation was used as a biocatalyst for the oxidation of NH⁺₄ to NO^?₂. Characterization of this immobilized biocatalyst was done according to the guidelines recommended by the Working Party on Immobilized Biocatalysts of the European Federation of Biotechnology. The most important indications obtained from the results are: (a) at low concentrations of substrate, either ammonium ions or oxygen, diffusion limitation will play a role; (b) inhibition by nitrite ions accumulating in the support is not rapidly controlling the efficiency of the immobilized cells; (c) accumulation of hydrogen ions is a rate-limiting factor, especially in unbuffered solutions; (d) the activity of immobilized N. europaea can increase as a result of growth in the support under conditions which would cause washout of free cells. This last result shows the potential of immobilized N. europaea for nitrification of wastewater. The development of a system applying a cheaper and more stable support is, however, a prerequisite for this application.     

Mammary carcinogenesis in different rat strains after irradiation and hormone administration

Radiation carcinogenesis of the rat mammary gland was investigated with the objective of investigating the combined effect of oestrogen administration and irradiation. Three rat strains, Sprague-Dawley, Wistar WAG/Rij and Brown Norway, with different susceptibilities to the induction of mammary cancer, have been irradiated with X-rays and mono-energetic neutrons. Increased hormone levels were obtained by subcutaneous implantation of pellets with oestradiol-17 beta (E2). The tumour incidence results were corrected for competing risks and were analysed with a continuous failure time distribution. The latency period for the hormone-treated animals is considerably shorter than for animals with normal endocrinological levels. Administration of the hormone results in an appreciable increase in the proportion of rats with malignant tumours. At the level of hormone administration applied in this study, radiation and hormones appear to produce an additive effect. The effect of hormone administration and irradiation for mammary tumourigenesis is equal for hormone administration one week prior to, or 12 weeks after irradiation. The RBE values for induction of mammary carcinomas after irradiation with 0.5 MeV neutrons have a maximum value of 20 and are not strongly dependent on the hormone levels.     

Effects of laundry wastewater on benthic algae in ditches in the Netherlands

Summary A modified, artificial substrate method was used to study the effects of laundry wastewater on benthic algae in a polder in the western part of the Netherlands. In a polluted ditch a clear chemical gradient was found with regard to the parameters conductivity, B.O.D., C.O.D., NH₄, ortho-P, total-P and Cl, showing decreasing values with increasing distance from the polluting source. pH and oxygen values showed the opposite. Algae, such asEuglena viridis, Nitzschia palea andOscillatoria spp. were positively correlated with the amount of pollution, while others, such asChrysochaete brittannica, Chrysophaera gallica, Cocconeis placentula andSynedra ulna showed a negative correlation with the amount of pollution. The results of algal counts are evaluated by means of diffenent methods,e.g. similarity coefficients, species abundance curves, diversity and saprobity indices. In different seasons benthic algal biomass proved to be a good indicator of pollution, except very close to the polluting source. The role of algal periodicity, influencing species composition, diversity and saprobity values, and biomass growth, has been discussed. The need of revising the saprobity system for stagnant ditches in the Netherlands, using artificial substrates, has been stressed.     

Habitat heterogeneity affects predation of European pine sawfly cocoons

Habitat heterogeneity is thought to affect top‐down control of herbivorous insects and contribute to population stability by providing a more attractive microhabitat for natural enemies, potentially leading to reduced population fluctuations. Identifying the parameters that contribute to habitat heterogeneity promoting top‐down control of herbivorous insects by natural enemies could facilitate appropriate management decisions, resulting in a decreased risk of pest insect outbreaks because of a higher level of predation. In our study, we measured the top‐down pressure exerted by small mammals on the cocoons of a notorious pest insect in pine forests, the European pine sawfly (Neodiprion sertifer), which is known to be regulated by small mammal predation. The forest stands used differed in heterogeneity measured in terms of differences in tree diversity and density, understory vegetation height, presence/absence, and density of dead wood. We found higher predation in more dense spots within forest stands. Further, the effect of dead wood on sawfly cocoon predation depended on the pine proportion in forest stands. The addition of dead wood in a manipulation experiment had a slight positive effect on cocoon predation, while dead wood removal caused a clear decrease in predation rate, and the decrease was more pronounced when the proportion of pine increased. Our results show that habitat heterogeneity affects predation by generalist predators on herbivorous insects. This knowledge could be applied to reduce the risk of insect outbreaks by applying management methods that increase heterogeneity in perennial systems such as forests and orchards, thus decreasing the levels of insect damage.     

Kinetic comparisons of mesophilic and thermophilic aerobic biomass

Kinetic parameters describing growth and decay of mesophilic (30°C) and thermophilic (55°C) aerobic biomass were determined in continuous and batch experiments by using oxygen uptake rate measurements. Biomass was cultivated on a single soluble substrate (acetate) in a mineral medium. The intrinsic maximum growth rate (μ _max) at 55°C was 0.71±0.09 h⁻¹, which is 1.5 times higher than the μ _max at 30°C (0.48±0.11 h⁻¹). The biomass decay rates increased from 0.004 h⁻¹ at 30°C to 0.017 h⁻¹ at 55°C. Monod constants were very low for both types of biomass: 9±2 mg chemical oxygen demand (COD) l⁻¹at 30°C and 3±2 mg COD l⁻¹at 55°C. Theoretical biomass yields were similar at 30 and 55°C: 0.5 g biomass COD (g acetate COD)⁻¹. The observed biomass yields decreased under both temperature conditions as a function of the cell residence time. Under thermophilic conditions, this effect was more pronounced due to the higher decay rates, resulting in lower biomass production at 55°C compared to 30°C. Electronic Publication     

Development of a test method to assess the sludge reduction potential of aquatic organisms in activated sludge.

This article shows the development of a quantitative sludge reduction test method, which uses the sludge consuming aquatic worm Lumbriculus variegatus (Oligochaeta, Lumbriculidae). Essential for the test are sufficient oxygen supply and the presence of a non-stirred layer of sludge for burrowing of the organisms. The test eliminates the unwanted effects of the organisms' movements, so-called bioturbation, on oxygen transport and (therefore) on sludge reduction. We used fresh untreated activated sludge grown on sewage, in order to stay close to the daily practice of sludge treatment. By separating sludge and worms, sludge reduction and worm growth are quantified independently and accurately. Predation by L. variegatus approximately doubles the decay rate of activated sludge. A minimum ratio of initial worm to sludge biomass (W(0)/S(0)) of about 0.4g worm/g sludge dry mass is required. Under the test conditions 20-40% of the predated sludge is converted into worm biomass. Our test is simple, reproducible and accurate and is done with equipment generally available in any laboratory, yielding results within a few days. The test can also be used to assess the application of mixtures of different aquatic organisms, but does not provide enough information for the design of a sludge treatment reactor.     

Apoptosis and p53 expression in rat adjuvant arthritis

The kinetics of apoptosis and the apoptosis-regulating gene p53 in adjuvant arthritis (AA) were investigated to assess the value of the AA rat model for testing apoptosis-inducing therapies. Very few terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end-labeling (TUNEL)-positive cells were detected during the early phases of AA, but on day 23 (chronic arthritis) the percentage of TUNEL-positive cells was significantly increased. Expression of p53 in synovial tissue gradually increased from days 5-23, which was markedly higher than p53 levels in rheumatoid arthritis (RA) synovium. Significant apoptosis only occurs late in rat AA and is concordant with marked p53 overexpression, making it useful model for testing proapoptotic therapies, but rat AA is not the best model for p53 gene therapy because dramatic p53 overexpression occurs in the latter stages of the disease.