Comparison of approaches towards ecotoxicity evaluation for the application of dredged sediment on soil

Purpose

The toxicity of 36 dredged sediments from the Czech Republic was investigated using a large battery of bioassays. The aim of this study was to investigate the feasibility of ecotoxicity testing in general and of individual bioassays more specific and to investigate how the results of bioassays are determined by the physicochemical properties of sediment samples and/or sediment contamination.

Material and methods

In 2008 and 2010, 36 sediment samples were collected from rivers and ponds and from sediment heaps in different parts of the Czech Republic. Both their physicochemical properties and their levels of contamination with POPs and heavy metals were analyzed. The ecotoxicities of the sediments were evaluated using the four bioassays from the new Czech directive 257/2009 Coll. concerning the application of dredged sediments on agricultural land (Enchytraeus crypticus reproduction, Folsomia candida reproduction, Lactuca sativa root elongation, and potential ammonium oxidation). The results of the four directive bioassays were compared with the results of other soil bioassays (Caenorhabditis elegans mortality, Eisenia fetida avoidance and reproduction) and eluate bioassays (Daphnia magna immobilization, Pseudokirchneriella subcapitata growth inhibition test, and Vibrio fischeri luminescence).

Results and discussion

We demonstrate that the battery suggested in Czech directive 257/2009 Coll. is highly effective in identifying toxic samples; these bioassays clearly revealing different types of toxicity and different exposure routes. Shorter alternative bioassays may be added especially when fast toxicity identification is needed. Eluate bioassays identified samples potentially hazardous to aquatic ecosystems. Their inclusion into the assessment scheme should be considered if the goal of assessment is also the protection of aquatic ecosystems. The results of our multivariate analysis show that specific physicochemical properties and contamination may affect bioassay responses. C. elegans was the most sensitive bioassay to physicochemical properties and also to organic contamination, while eluate bioassays were sensitive to heavy metal pollution.

Conclusions

Most effects detected by the bioassays could not be explained by the levels of toxicants measured or by the natural characteristics of sediments. Our results show that bioassays are irreplaceable in dredged sediment risk assessment because they complement information provided by chemical analyses.     

Software for estimating reference evapotranspiration using limited weather data

The FAO-56 Penman-Monteith combination equation (FAO-56 PM) has been recommended as the standard equation for estimating reference evapotranspiration (ET₀). The FAO-56 PM equation requires the numerous weather data that are not available in the most of the stations. The main goal of this paper is to present the software for estimating reference evapotranspiration, focusing on the feature of using limited weather data. This is simple Windows-based and user-friendly software provides methods to estimate extra-terrestrial radiation, maximum sunshine hours, daily net radiation and daily/monthly ET₀. The program is written in C# and includes comprehensive technical documentation. The software is available for free download.The weather data for this study were obtained from CIMIS for Davis weather station. The reduced-set FAO-56 PM approaches and adjusted Hargreaves equation were compared to the full-set FAO-56 PM equation. The FAO-56 reduced-set PM ET₀ estimates were in closest agreement with FAO-56 full-set PM ET₀ estimates. The adjusted Hargreaves equation (AHARG) was found to be in very good agreement with the full-set FAO-56 PM. This program is the first software facilitating calculation of ET₀ only with air temperature parameter.     

Genetic variability of <Emphasis Type="Italic">Plum pox virus</Emphasis> isolates in the Czech Republic

Plum pox virus (PPV), the causal agent of Sharka disease, is an important pathogen of stone fruit trees. In this study, 24 new Czech PPV isolates from five different orchards were collected and characterized, molecularly. PPV-D isolates were identified in all orchards studied; whereas PPV-Rec isolates were identified in only two of them. A phylogenetic analysis on (Cter) NIb-(Nter) CP was performed. Three Czech PPV-D isolates BOH11CZ, BOH12CZ, and BOH13CZ diverged into a significantly separated cluster. PPV-Rec isolates formed a fairly homogenous group. However, the Bohutice and the Lipov PPV-Rec isolates clustered in two significantly separated branches.     

Evaluation of a serodiagnostic test using Ascaris suum haemoglobin for the detection of roundworm infections in pig populations

The significant economical consequences of infections with Ascaris suum in pigs are already well documented. However, due to the subclinical nature of the disease and the lack of practical diagnostic means, ascariasis often remains undiagnosed. Here we describe the development and evaluation of a novel indirect ELISA using the purified A. suum haemoglobin (AsHb) molecule as an antigen. Initial validation using sera from 190 pigs experimentally infected twice a week with A. suum and Trichuris suis (25 and 5eggskg(-1)day(-1) respectively) demonstrated that the AsHb ELISA is able to detect long-term exposure to A. suum with a high sensitivity and specificity (99.5% and 100.0% respectively). Furthermore, this serological technique proved to be more sensitive than faecal examination on week 7 and 14 of the experiment (99.5% and 100% compared to 59.5% and 68.4% respectively). Cross-reactivity caused by T. suis infection was shown to be limited after analysing sera from pigs with an experimental T. suis mono-infection. Seroconversion was shown to occur from week 6 onwards in pigs receiving 100 A. suum eggs 5 times a week. Preliminary testing of the ELISA on six randomly selected farms confirmed the results obtained in the artificial infection trials, showing a higher sensitivity of the serologic method compared to faecal examination. Finally, the ELISA was used to investigate Ascaris infection rates on 101 conventional Flemish pig farms. The results showed that on 38.6% of the farms less than 20% of the tested samples were seropositive, while in 19.8% of the farms 80-100% of all pigs were seropositive. The results of this study suggest that the AsHb ELISA could provide pig farmers and veterinarians with an easier and more sensitive way to estimate the overall prevalence of A. suum on their farm.     

Pre‐slaughter mortality among turkeys related to their transport

Overall mortality among turkeys transported for slaughter in the Czech Republic from 2009 to 2014 was 0.147%. The lowest mortality (0.023%) was found in turkeys transported for distances up to 50 km; longer distances were associated with increasing death rates, with the highest losses (0.543%) recorded for distances from 201 to 300 km. Differences were also found when comparing transport‐related mortality rates according to the season of the year. The highest mortality (0.228%) was associated with transports carried out in summer, whereas the lowest death rates were found in turkeys transported for slaughter in winter (0.105%) and autumn (0.113%). According to the ambient temperatures in which transports associated with the highest death rates among turkeys were carried out (14 to 21°C), it appears that heat stress can be a major factor in the birds’ mortality. However, low ambient temperatures (i.e. below ?2°C) also appear to increase transport‐related mortality, as shown by our results. In addition to shortening the lengths of transport distances, appropriate consideration of prevailing climatic conditions and the sensitivity of turkeys to high and low temperatures during transport may also lead to a reduction in the number of turkeys dying during transport for slaughter.     

ELISA for sulfonamides and its application for screening in water contamination

Two enzyme-linked immunosorbent assays (ELISAs) were tested for their suitability for detecting sulfonamides in wastewater from various stages in wastewater treatment plants (WWTPs), the river into which the wastewater is discharged, and two swine-rearing facilities. The sulfamethoxazole ELISA cross-reacts with several compounds, achieving detection limits of <0.04 microg/L for sulfamethoxazole (SMX), sulfamethoxypyridine, sulfachloropyridine, and sulfamethoxine, whereas the sulfamethazine (SMZ) ELISA is more compound specific, with a detection limit of <0.03 microg/L. Samples from various stages of wastewater purifications gave 0.6-3.1 microg/L by SMX-ELISA, whereas river samples were approximately 10-fold lower, ranging from below detection to 0.09 microg/L. Swine wastewater samples analyzed by the SMX-ELISA were either at or near detectable limits from one facility, whereas the other facility had concentrations of approximately 0.5 microg/L, although LC-MS/MS did not confirm the presence of SMX. Sulfamethazine ELISA detected no SMZ in either WWTP or river samples. In contrast, wastewater samples from swine facilities analyzed by SMZ-ELISA were found to contain approximately 30 microg/L [piglet (50-100 lb) wastewater] and approximately 7 microg/L (market-weight hog wastewater). Sulfamethazine ELISA analyses of wastewater from another swine facility found concentrations to be near or below detection limits. A solid phase extraction method was used to isolate and concentrate sulfonamides from water samples prior to LC-MS/MS multiresidue confirmatory analysis. The recoveries at 1 microg/L fortification ranged from 42 +/- 4% for SMZ to 88 +/- 4% for SMX ( n = 6). The ELISA results in the WWTPs were confirmed by LC-MS/MS, as sulfonamide multiresidue confirmatory analysis identified SMX, sulfapyridine, and sulfasalazine to be present in the wastewater. Sulfamethazine presence at one swine-rearing facility was also confirmed by LC-MS/MS, demonstrating the usefulness of the ELISA technique as a rapid and high-throughput screening method.     

Cultivation of flax in spoil‐bank clay: Mycorrhizal inoculation vs. high organic amendments

A greenhouse experiment was focused on the application of arbuscular mycorrhizal fungi (AMF) in effective crop production during reclamation of coal‐mine spoil banks. The aim of the study was to find out whether mycorrhizal inoculation improves growth of flax (Linum usitatissimum L.) and whether it can compensate for high doses of expensive organic amendment. Flax was planted in original spoil‐bank gray Miocene clay amended with organic matter used for spoil‐bank reclamation (mixture of composted urban waste and lignocellulose papermill waste). This amendment was applied in four descending doses equivalent to the application of 500, 200, 100, and 0 t ha^–1. The plants received either a mixed inoculum of three AMF species (Glomus mosseae, G. claroideum, and G. intraradices) or were left uninoculated. Growth of flax was significantly increased by mycorrhizal inoculation in pure clay (by 60%) as well as in clay with all amendment doses (by 119% on average). Also, the addition of organic matter increased plant growth but, interestingly, the biomass production was comparable regardless the dose of amendment. Our results suggest that similar yields are attainable with only one fifth of the usual fertilization dose, which can significantly decrease costs related to the reclamation of spoil banks. If mycorrhizal inoculation is applied together with the optimized fertilization, growth of flax planted in spoil‐bank clay can be further improved.