Evaluation of frameworks for ecotoxicological hazard classification of waste.

A new harmonized EU regulation for the classification of waste came into effect on 1st June 2015, in which the criteria and assessment methods for the classification of hazardous waste are harmonized with other internationally agreed-upon systems for hazard classification of chemicals (CLP). However, criteria and guidance for the assessment of ecotoxicological hazard (Hazard Property 14, HP14) are still lacking for waste classification. This paper have evaluated and compared two HP14 classification frameworks: (i) a calculation method (summation) for mixtures, and (ii) leaching tests. The two frameworks were evaluated by surveying and evaluating ecotoxicological data for Cu, Zn, K and Ca species in bottom ash from incinerated waste, together with geochemical speciation modelling. Classification based on the summation method proved to be highly sensitive to the choice of speciation and ecotoxicological classification. This results in a wide range of critical concentrations triggering hazardous classification (in particular for Cu and Zn). Important parameters governing the availability of toxic elements, such as transformation from one species to another and complexation on organic or inorganic sorbents, are not accounted for. Geochemical modelling revealed that a testing strategy built on CLP based leaching tests (liquid/solid ratio (L/S)⩾10,000, pH range 5.5-8.5) avoids bias and is superior to the summation method with respect to both precision and accuracy. A testing strategy built on leaching tests, designed for risk assessment purposes, (L/S ratio of 10, natural pH of the ash) severely underestimate the hazard associated with the presence of toxic compounds (Cu and Zn), while simultaneously falsely indicate a hazardousness due to the presence of non-toxic compounds (Ca and K). However, the testing methods adopted by CLP are problematic from a practical and functional point of view. To conclude, the L/S ratio and pH were found to be critical for hazard classification based on leaching test methods. Further studies are needed to develop a relevant, practical and functional testing strategy for HP14 hazardous waste classification.

Influence of leaching conditions for ecotoxicological classification of ash.

The Waste Framework Directive (WFD; 2008/98/EC) states that classification of hazardous ecotoxicological properties of wastes (i.e. criteria H-14), should be based on the Community legislation on chemicals (i.e. CLP Regulation 1272/2008). However, harmonizing the waste and chemical classification may involve drastic changes related to choice of leaching tests as compared to e.g. the current European standard for ecotoxic characterization of waste (CEN 14735). The primary aim of the present study was therefore to evaluate the influence of leaching conditions, i.e. pH (inherent pH (∼10), and 7), liquid to solid (L/S) ratio (10 and 1000 L/kg) and particle size (<4 mm, <1 mm, and <0.125 mm), for subsequent chemical analysis and ecotoxicity testing in relation to classification of municipal waste incineration bottom ash. The hazard potential, based on either comparisons between element levels in leachate and literature toxicity data or ecotoxicity testing of the leachates, was overall significantly higher at low particle size (<0.125 mm) as compared to particle fractions <1mm and <4mm, at pH 10 as compared to pH 7, and at L/S 10 as compared to L/S 1000. These results show that the choice of leaching conditions is crucial for H-14 classification of ash and must be carefully considered in deciding on future guidance procedures in Europe.

An ecotoxicological evaluation of aged bottom ash for use in constructions.

Municipal and Industrial Solid Waste Incineration (MISWI) bottom ash is mainly deposited in landfills, but natural resources and energy could be saved if these ash materials would be used in geotechnical constructions. To enable such usage, knowledge is needed on their potential environmental impact. The aim of this study was to evaluate the ecotoxicity of leachates from MISWI bottom ash, aged for five years, in an environmental relevant way using a sequential batch leaching method at the Liquid/Solid-ratio interval 1-3, and to test the leachates in a (sub)chronic ecotoxicity test. Also, the leachates were characterized chemically and with the technique of diffusive gradients in thin films (DGTs). By comparing established ecotoxicity data for each element with chemically analysed and labile concentrations in the leachates, potentially problematic elements were identified by calculating Hazard Quotients (HQ). Overall, our results show that the ecotoxicity was in general low and decreased with increased leaching. A strong correspondence between calculated HQs and observed toxicity over the full L/S range was observed for K. However, K will likely not be problematic from a long-term environmental perspective when using the ash, since it is a naturally occurring essential macro element which is not classified as ecotoxic in the chemical legislation. Although Cu was measured in total concentrations close to where a toxic response is expected, even at L/S 3, the DGT-analysis showed that less than 50% was present in a labile fraction, indicating that Cu is complexed by organic ligands which reduce its bioavailability.

Prevailing effectiveness of the 2009 influenza A(H1N1)pdm09 vaccine during the 2010/11 season in Sweden.

Sixty per cent of the Swedish population received the monovalent AS03-adjuvanted pandemic influenza vaccine in the autumn of 2009. We assessed the age-specific effectiveness of this pandemic vaccine against hospitalisation with laboratory-confirmed influenza A(H1N1)pdm09 during the season 2010/11, in the age group from six months to 64 years in Sweden. The screening method was applied to available surveillance data. Our results suggest a prevailing effectiveness of 72% (95% confidence interval (CI): 63­80%) with the highest effectiveness among children, six months to nine years-old (92%, 95%CI: 80­97%). However, there were limitations in data quality and study design due to the lack of systematic recording of administered vaccinations, which underline the importance of preparing for an evaluation when planning for large public health actions. Despite these limitations, we believe the results reflect true, high prevailing vaccine effectiveness. Indeed, there were fewer deaths caused by influenza and the impact of influenza on intensive care units was less severe during the 2010/11 season in Sweden than in countries with lower pandemic vaccination coverage. The association between the pandemic vaccine and narcolepsy has increased the importance of assessing the risks and benefits of the vaccination; studies on the effectiveness and the duration of protection are needed for this.

Improved understanding of key elements governing the toxicity of energy ash eluates.

Ash from incinerated waste consists mainly of a complex mixture of metals and other inorganic elements and should be classified based on its inherent hazardous effects according to EUs Waste Framework Directive. In a previous study, we classified eight eluates from ash materials from Swedish incineration plants, both chemically and ecotoxicologically (using bacteria, algae, crustacean and fish). Based on measured concentrations in the eluates together with literature acute toxicity data on the crustacean Nitocra spinipes we identified six elements (i.e. Zn, Cu, Pb, Al, K and Ca) potentially responsible for the observed ecotoxicity. However, comparing the used test methods with N. spinipes, the acute test was relatively insensitive to the eluates, whereas the (sub)chronic test (i.e. a partial life cycle test, investigating larval development ratio) was very sensitive. The overall aim of this follow-up study was to verify if the pinpointed elements could be responsible for the observed (sub)chronic toxicity of the eluates. Individual effect levels (i.e. NOEC values) for these six elements were therefore generated using the (sub)chronic test. Our results show that for six of the eight eluates, the observed ecotoxicity can be explained by individual elements not classified as ecotoxic (Al, K and Ca) according to chemical legislation. These elements will not be considered using summation models on elements classified as ecotoxic in solid material for the classification of H-14, but will have significant implications using ecotoxicological test methods for this purpose.

An ecotoxicological approach for hazard identification of energy ash.

Within the EU, ash should be classified by its inherent hazardous effects under criterion H-14 (ecotoxic) in the Directive on waste (2008/98/EC). Today, however, there are no harmonized quantitative criterions for such a classification, but it is stated that biological test systems can be used. In this study seven ash materials were leached and characterized, both biologically and chemically. The objectives were to evaluate if (a) clear concentration-response relationships could be achieved for the selected toxicity tests (bacteria, algae, crustacean and fish), (b) some test(s) are generally more sensitive and (c) the toxic responses were consistent with the chemical analyzes. Interestingly, our results indicate that high concentrations of non-hazardous components (Ca, K) influenced the toxicity of almost all ash eluates, whereas hazardous components (e.g. Zn, Pb) only influenced the toxicity of the eluates ranked as most hazardous. If considering both hazardous and non-hazardous substances, the observed toxic responses were relatively consistent with the chemical analyzes. Our results further showed that the (sub)chronic tests were much more sensitive than the acute tests. However, the use of extrapolation factors to compensate for using the less sensitive acute tests will likely lead to either over- or underestimations of toxicity. Our recommendation is therefore that classification of waste according to H-14 should be based on (sub)chronic test data. Finally, given that treatment of the eluates prior to toxicity testing has a major significance on the concentration and speciation of released substances, further studies are needed in order to propose a relevant testing scheme.

Fibrin glue (Tisseel) added with sodium hyaluronate in microvascular anastomosing.

To test the effect on the handling properties of a two component fibrin glue, the viscosity was increased with sodium hyaluronate and the glue was applied to a microvascular anastomosis in rats. The femoral artery of each rat was anastomosed with three conventional sutures and then sealed with fibrin glue. Three glues with different viscosities were tested: original Tisseel fibrin glue (Immuno AG, Vienna); Tisseel with 0.9% sodium chloride added to the fibrinogen component; and Tisseel with a high molecular weight sodium hyaluronate (10 mg/ml, Healon, Pharmacia, Sweden) added to the fibrinogen component. The increased viscosity of the fibrin glue to which hyaluronate had been added resulted in a significantly higher patency rate 20 minutes after completion of the anastomosis (p < 0.01), and reduced the amount of fibrin that entered the vessels.

Determination of biomechanical characteristics of restrictive adhesions and of functional impairment after flexor tendon surgery: a methodological study of rabbits.

Formation of restrictive adhesions is one of the main obstacles in rehabilitation following hand surgery. Most experimental work, however, involves only a macroscopic and/or histologic evaluation of the amount of adhesions, and their functional characteristics are poorly described. The aim of this study was to develop an experimental technique for characterization of the biomechanical properties of the finger-tendon unit. An instrument was developed for continuous and simultaneous recording of tensile load, tendon excursion and angular rotation in the distal interphalangeal joint of rabbit digits. Utilizing this instrument, it was revealed that the first 50 degrees of flexion required virtually no tensile load either in unoperated digits or immediately after tenorrhaphy. Thereafter, the load required to obtain further flexion was progressively increased. The strength of adhesions, determined 2 weeks after tenorrhaphy, was best expressed as the maximum tensile load recorded before 50 degrees of flexion was reached. This measurement could also be used to register the strength of the tendon repair and to detect partial tendon rupture during the measurement. The technique allows both adequate measurements of the strength of the adhesions and of the tendon gliding ability after flexor tendon surgery.

An in vivo method for studies of traumatic vasospasm.

A study was undertaken to develop a model for investigations of traumatically induced vasospasm. The left ear of the rabbit was denervated under short anesthesia. With the animal under light sedation, spasm of the central ear artery was repeatedly induced with a clip applying forceps. To evaluate the vasospasm, the vessel was transilluminated with cold light, and the internal diameter was continuously measured. The spasm was assessed in terms of its duration, intensity (reduction of initial diameter) and severity (integrated change in diameter over time). Repetitive application of the forceps on the same vessel segment shortened the spasm duration. With prolonged pinch duration, the spasm duration was lengthened. The preparation was stable for at least 3 hours. This model was effective for manipulating small vessels and producing spasm and is of potential value for studying the treatment of vasospasm by topical local intra-arterial and systemic methods.

A self-migration method for preparation of sperm for in-vitro fertilization.

Human sperm samples (n = 211) were prepared for in-vitro fertilization (IVF) and embryo transfer by a self-migration procedure in Earle's medium containing highly purified hyaluronic acid (Hya) (MW 3,000,000) included to increase the viscosity of the medium. The method resulted in the recovery of a significantly higher percentage of motile spermatozoa compared with the traditional centrifugation method, 87.5 +/- 0.9% versus 76.1 +/- 1.3% (P less than 0.001). When comparing media with and without Hya in the self-migration method for preparation of normal sperm samples, the media containing Hya resulted in the recovery of a significantly higher percentage of motile spermatozoa, 89.0 +/- 0.8% versus 73.8 +/- 2.0% (P less than 0.001). In a group of 80 consecutive couples entering our IVF programme, sperm samples from 44 of the men were allocated at random for the self migration method in medium containing Hya and sperm samples from 36 men for preparation by centrifugation and swim-up. Significantly more pregnancies were achieved in the group prepared in medium containing Hya. It is concluded that self-migration of sperm in a medium containing Hya is simple and rapid, and results in a high recovery of motile spermatozoa which can be used for in-vitro insemination of human oocytes with favourable results.

Rheology of hyaluronate.

Solutions containing high molecular weight hyaluronate at concentrations around 10 mg/ml exhibit interesting rheological properties due to formation of a highly entangled network of flexible polysaccharide molecules. We have performed an extensive study of the rheological properties of hyaluronate solutions as a function of concentration and molecular weight. In this paper we review some basic rheological concepts, and discuss the rheological properties of hyaluronate solutions at high concentrations and medium to high molecular weights (1-5 million). The bulk viscosity (zero shear viscosity) of hyaluronate solutions is strongly dependent both on concentration and molecular weight. A 2-fold increase in concentration or molecular weight results in a 10-fold increase in bulk viscosity. For application in body compartments, the concentration of hyaluronate cannot be increased much above 10 mg/ml due to the highly non-ideal colloid osmotic behaviour of hyaluronate. High viscosity hyaluronate solutions must therefore be based on high molecular weight material.

Shear flow characteristics of sodium hyaluronate. Relationship to performance in anterior segment surgery.

In this study, methods were developed for the in vitro evaluation of the surgical performance characteristics of viscoelastic fluids, such as sodium hyaluronate (Healon). Sodium hyaluronate exhibited superior surgical performance to chondroitin-6-sulfate. The superior performance of sodium hyaluronate resulted in part from its high viscosity, which is 20 times greater than that of chondroitin-6-sulfate at shear rates on the order of 10 s-1. The gel-like character of sodium hyaluronate as evidenced by the creep flow behavior, was greater than that of chondroitin-6-sulfate and was important for maintaining depth in the anterior chamber. It was shown that a threshold of 80 poise for the shear viscosity (at approximately 10 s-1 was needed for useful performance in surgery. It was also shown that 0.42 USP units of hyaluronidase per 1.0 mg sodium hyaluronate produces a 90% decrease in the shear viscosity (at approximately 10 s-1) within approximately 2 1/2 hours. Use of sodium hyaluronate in conjunction with hyaluronidase would allow sodium hyaluronate to remain highly viscous during surgery, but would gradually become less viscous to facilitate aqueous outflow after surgery.

Intra-articular injection of high-molecular hyaluronic acid. An experimental study on normal adult rabbit knee joints.

High-molecular hyaluronic acid was injected repeatedly into normal knee joints of adult rabbits. Histologically the articular cartilage was not affected by this treatment. The content of chondroitin sulphate, assessed by a histochemical method, was not altered and neither was the water content of the hyaline articular cartilage. On the other hand, in the synovial membrane of joints treated with hyaluronic acid a diffuse infiltration of plasma cells and lymphocytes was observed, indicating that the hyaluronic acid administered exerts its action via the synovial membrane.