Capillary and venous Babesia canis rossi parasitaemias and their association with outcome of infection and circulatory compromise

This observational study of 100 dogs naturally infected with Babesia canis rossi determined whether severity of parasitaemia was associated with outcome of infection and documented the relative distribution of parasitised red blood cells (pRBC) in capillary and venous circulation. The association between increased parasitaemias and outcome with a clinically compromised circulation was also investigated. Outcome was defined as either hospitalisation with death, or hospitalisation with eventual recovery or treatment as an outpatient. Dogs were enrolled if large babesias were found on stained thin capillary blood smears made from an ear prick. Thin venous smears were prepared from jugular or cephalic blood. Parasitaemias were manually counted and expressed as the percent pRBC. Ten dogs died, 50 recovered after hospitalisation and 40 were treated as outpatients. Venous sampling site did not affect venous parasitaemia (P=0.6). Both capillary and venous parasitaemias of dogs that died were significantly higher than those of dogs that recovered after hospitalisation (P=0.002) and dogs that were treated as outpatients (P<0.0001). When assessing the whole group, capillary parasitaemia (median 0.61%, range <0.05-71.6%, interquartile range (IQR) 0.22-3.75%) was significantly higher than venous parasitaemia (median 0.14%, range 0-30.6%, IQR 0.046-0.52%) with P<0.0001. The 21 dogs with a clinically compromised circulation were more likely to die (P<0.0001) and had significantly higher capillary (median 5.98%, range 0.09-71.6%, IQR 2.44-19.41%) and venous (median 2.81%, range <0.05-30.6%, IQR 0.17-9.03%) parasitaemias than the 79 dogs with a clinically normal circulation (capillary median parasitaemia 0.38%, range <0.05-12.87%, IQR 0.16-1.42%; venous median parasitaemia 0.096%, range 0-6.13%, IQR <0.05-0.33%; P<0.0001). This study shows that high parasitaemia is significantly associated with death in B c rossi infected dogs. The previous clinical suspicion that capillary parasitaemias are usually higher than venous parasitaemias is confirmed. Thus capillary samples are the most appropriate diagnostic samples. Prior observations that a clinically compromised circulation is associated with death are confirmed. Despite the highly significant association between compromised circulation and higher parasitaemia, it is thought unlikely that parasite burden is the sole trigger for circulatory collapse.     

Loss of low molecular weight dissolved organic carbon (DOC) and nitrogen (DON) in H2O and 0.5 M K2SO4 soil extracts

Soil extracts are routinely used to quantify dissolved organic nutrient concentrations in soil. Here we studied the loss and transformation of low molecular weight (LMW) components of DOC (¹⁴C-glucose, 1 and 100 μM) and DON (¹⁴C-amino acid mixture, 1 and 100 μM) during extraction of soil (0-6 h) with either distilled water or 0.5 M K₂SO₄. The extractions were performed at 20 °C, at 4 °C, or in the presence of an inhibitor of microbial activity (HgCl₂ and Na-azide). We showed that both glucose and amino acids became progressively lost from solution with increasing shaking time. The greatest loss was observed in H₂O extracts at 1 μM for both substances (>90% loss after 15 min). Lower temperature (4 °C) and presence of K₂SO₄ both resulted in reduced loss rates. The presence of microbial inhibitors effectively eliminated the loss of glucose and amino acids. We conclude that microbial transformation of LMW-DOC and DON during H₂O or K₂SO₄ extraction of soil may affect the estimation of their concentrations in soil. This finding has significant implications for methods that rely on chemical extractions to estimate LMW-C components of DOC and DON.     

Atmospheric deposition to remote receptors III. Statistical analysis of precipitation data from the ILWAS-network

A statistical analysis of the precipitation data base from the Integrated Lake-Watershed Acidification Study (ILWAS) is presented. Included in this analysis are: equivalence of distributional form of the concentrations, ion pair concentration analysis, regression by site and season, precipitation type, and multiple regressions. The analysis emphasizes the ions: H, NH₄, SO₄, and NO₃. Variability in H is explained largely through SO₄ and NO₃; spatial variability for the species pairs SO₄-H, NO₃-H, SO₄-NH₄; and NO₃-NH₄ does not suggest significant differences between sites on a seasonal basis.     

Modeling of throughfall chemistry and indirect measurement of dry deposition

Precipitation over a forested watershed is altered by interaction with plant surfaces which act as a filter for airborne gases and particles. This results in a major transfer to the forest floor of materials captured, washed, and leached from the forest canopy. Sequential samples of wetfall and sequential samples of throughfall under deciduous and coniferous trees were collected and chemically analyzed for major anions and cations. A simple washoff, mixing model based on leaf area index was used to simulate throughfall chemistry and to decouple foliar exudation from dry deposition. Model results gave excellent predictions of the measured sequential throughfall using estimated values of dry deposition. The model can also be used to calculate dry deposition, if the sequential throughfall data and wetfall data are used as input variables.     

Agricultural suitability of degraded Acrisols and Lixisols of former tea lands in Sri Lanka

Physical and chemical soil degradation hinder the introduction of mixed cropping systems on former tea lands in the Central Highlands of Sri Lanka. For instance on two typical soil catenas soil properties such as texture, pH, acidity parameters, pedogenic oxides and available plant nutrients indicating fertility are presented. Obviously Al³⁺ occupies a larger part of the exchange complex and accounts for more soil acidity than H⁺. Al toxicity is hence a major restricting factor for plant growth in the area. The investigated data also permit an evaluation of soil erodibility and a soil fertility assessment according to the Fertility Capability Classification System (Sanchez et al., 1982). Agricultural measures to reduce soil acidity and cover the lime requirements of the soils are advised. Additional soil conservation measures should diminish soil erosion in order to improve the sustainability of the cropping systems.     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Cross-modality of texture and aroma perception is independent of orthonasal or retronasal stimulation

To assess the influence of orthonasal and retronasal stimulation on cross-modal interactions between texture and flavor perception of food, a series of experiments have been conducted. Healthy human subjects were exposed to strawberry aroma pulses delivered by a computer-controlled stimulator based on air dilution olfactometry. Just prior to exposure to the aroma, the human subjects consumed water, custard, or protein gels with different textures without any added aroma. The aroma was delivered as a sequence of aroma pulses, in either an orthonasal or a retronasal fashion. The retronasal presentation of aroma with concomitant presentation of texture is thought to more closely mimick the in vivo flavor release of semisolid food products as compared to orthonasal stimulation. The time between oral consumption of the food, including swallowing, and the exposure to the aroma varied between 0.5 and 6.5 s. The subjects rated the intensity of the strawberry aroma. It was observed that the intensity of aroma decreased with increasing firmness of the food that was consumed. Aroma pulses delivered 6.5 s after swallowing were perceived as being more intense as compared to aroma pulses delivered immediately after swallowing. In conjunction with late delivery, the effect cross-modal interactions apparently decreased. Significantly higher odor intensities were reported for the aroma stimuli supplied orthonasally in comparison to retronasal administration. The observed cross-modal effect of texture on aroma intensity was not significantly altered by the mode of aroma delivery, i.e., orthonasal or retronasal stimulus administration.