The use of bioassays for evaluating the toxicity of sewage sludge and sewage sludge-amended soil

Background and Aims  

In soils, the most commonly mentioned hazardous substances are metals. One of the sources of its accumulation is the application of sewage sludge. However, little information is available regarding the estimation of the toxicity of sewage sludge or soil treated with sewage sludge, even by means of a battery of bioassays. In this study an evaluation of a battery of bioassays was carried out for toxicity assessment of sewage sludge and sewage sludge-treated soil. The objectives of this study were a) to compare the sensitivity of the different bioassays for the toxicity determination of sewage sludge contaminated with metals and soil treated with this sewage sludge, b) to elaborate a procedure for the attribution of sewage sludge samples to hazard classes based on the ecotoxicological data, and c) to evaluate the suitability of elutriate bioassays and microbial toxicity tests for the assessment of sewage sludge-treated soil.     

New sources of resistance to race Ro1 of the Golden nematode (Globodera rostochiensis Woll.) among reputed duplicate germplasm accessions of Solanum tuberosum L. subsp. andigena (Juz. et Buk.) Hawkes in the VIR (Russian) and US Potato Genebanks

Cultivated Solanum tuberosum L. subsp. andigena is well known as a rich source of valuable traits for potato breeding, especially for resistance to diseases and pests. The golden potato cyst nematode, Globodera rostochiensis Woll., is considered to be one of todays most serious hindrances to potato production in Europe and North America. Thus, the breeding of new cultivars that have resistance to PCN is of great importance. The USPG (USA) and VIR (Russian) potato genebanks, as well as others, maintain many samples of primitive cultivated and wild potato species originating from Latin America. Many of these samples are assumed to be genetically duplicated because the material in both genebanks came from the same original source. A joint investigation of new genotypes of subsp. andigena forms resistant to potato cyst nematode (PCN) was carried out on samples of subsp. andigena at VIR with reputed duplicate samples at USPG. After careful screening, 14 samples which possessed resistance to PCN were identified. A high level of this resistance was transmitted to sexual progeny at a high frequency for all of the selections. Eleven of the accessions found to be resistant have reputed duplicates in USPG that were not previously known to be resistant. Thus, this research not only broadens the choice of parents available for resistance breeding, but identifies model materials for future research to test the parity of PCN resistance among reputed duplicate samples in the two genebanks.     

Effect of cooking on concentrations of β-estradiol and metabolites in model matrices and beef

Because beef food products are generally cooked prior to consumption, the behavior of chemicals in these cooked foods is important in estimating human exposure. The heat stability of the natural estrogen β-estradiol (β-E2) and its metabolites α-estradiol (α-E2), estrone (E1), and several catechol estrogens was examined in heated vegetable oil and aqueous solutions. The chemicals were also incorporated into regular and extra lean ground beef and subjected to cooking. E1 and E2 were stable in aqueous solutions at 100°C, whereas the catechol estrogens exhibited first-order decay curves with half-lives of 2-10 min. Their stability improved to the same level as the other test chemicals when an antioxidant was added to the solution, suggesting that their disappearance was due to oxidation rather than thermal degradation. E1 and E2 were also stable in heated vegetable oil (160-180°C), whereas catechol estrogen decreased 30-50% over the 2 h duration of the experiments. Chemical losses from cooked beef appear to be related to the fat content of the beef, with greater losses occurring in regular ground beef (25-30%), compared to extra lean ground beef (5-20%). This study shows that cooking reduces but does not eliminate the potential for dietary exposure to growth promoters in ground beef.     

Biochemical Properties and Anti-Biofilm Activity of Chitosan-Immobilized Papain

Chitosan, the product of chitin deacetylation, is an excellent candidate for enzyme immobilization purposes. Here we demonstrate that papain, an endolytic cysteine protease (EC: 3.4.22.2) from Carica papaya latex immobilized on the matrixes of medium molecular (200 kDa) and high molecular (350 kDa) weight chitosans exhibits anti-biofilm activity and increases the antimicrobials efficiency against biofilm-embedded bacteria. Immobilization in glycine buffer (pH 9.0) allowed adsorption up to 30% of the total protein (mg g chitosan⁻¹) and specific activity (U mg protein⁻¹), leading to the preservation of more than 90% of the initial total activity (U mL⁻¹). While optimal pH and temperature of the immobilized papain did not change, the immobilized enzyme exhibited elevated thermal stability and 6–7-fold longer half-life time in comparison with the soluble papain. While one-half of the total enzyme dissociates from both carriers in 24 h, this property could be used for wound-dressing materials design with dosed release of the enzyme to overcome the relatively high cytotoxicity of soluble papain. Our results indicate that both soluble and immobilized papain efficiently destroy biofilms formed by Staphylococcus aureus and Staphylococcus epidermidis. As a consequence, papain, both soluble and immobilized on medium molecular weight chitosan, is capable of potentiating the efficacy of antimicrobials against biofilm-embedded Staphylococci. Thus, papain immobilized on medium molecular weight chitosan appears a presumably beneficial agent for outer wound treatment for biofilms destruction, increasing antimicrobial treatment effectiveness.     

Functional Characterization of a New GH107 Endo-α-(1,4)-Fucoidanase from the Marine Bacterium Formosa haliotis

Fucoidans from brown macroalgae are sulfated fucose-rich polysaccharides, that have several beneficial biological activities, including anti-inflammatory and anti-tumor effects. Controlled enzymatic depolymerization of the fucoidan backbone can help produce homogeneous, defined fucoidan products for structure-function research and pharmaceutical uses. However, only a few endo-fucoidanases have been described. This article reports the genome-based discovery, recombinant expression in Escherichia coli, stabilization, and functional characterization of a new bacterial endo-α-(1,4)-fucoidanase, Fhf1, from Formosa haliotis. Fhf1 catalyzes the cleavage of α-(1,4)-glycosidic linkages in fucoidans built of alternating α-(1,3)-/α-(1,4)-linked l-fucopyranosyl sulfated at C2. The native Fhf1 is 1120 amino acids long and belongs to glycoside hydrolase (GH) family 107. Deletion of the signal peptide and a 470 amino acid long C-terminal stretch led to the recombinant expression of a robust, minimized enzyme, Fhf1Δ470 (71 kDa). Fhf1Δ470 has optimal activity at pH 8, 37–40 °C, can tolerate up to 500 mM NaCl, and requires the presence of divalent cations, either Ca²⁺, Mn²⁺, Zn²⁺ or Ni²⁺, for maximal activity. This new enzyme has the potential to serve the need for controlled enzymatic fucoidan depolymerization to produce bioactive sulfated fucoidan oligomers.     

Toxicity of biofungicide Timorex 66 EC to Cladobotryum dendroides and Agaricus bisporus

Twenty Cladobotryum dendroides isolates from Serbian Agaricus bisporus farms collected during 2003–2007, and the F56 strain of A. bisporus were tested in vitro for sensitivity to tea tree oil (Timorex 66 EC), a biofungicide, in comparison with prochloraz–manganese (Octave WP). The efficacies of tea tree oil and prochloraz–manganese were evaluated in a mushroom growing room, after application at standard product application rates and a combination of the two at respective proportion of 20:80%. Tea tree oil was considerably less toxic than prochloraz–manganese in vitro to C. dendroides isolates (ED₅₀ 112.9–335.8 mg l⁻¹) and A. bisporus F56 strain (98.0 mg l⁻¹), although neither fungicide was lethal to the pathogen. The biological efficiency of tea tree oil was higher than in treatments with the reference formulation of prochloraz–manganese. In our in vivo trials, no negative interference of the biofungicide with A. bisporus physiology was observed. Tea tree oil applied at the standard product application rate caused a significant reduction in cobweb disease levels in the A. bisporus growing room. Timorex 66 EC should be tested further in combination with other biofungicides to investigate the effectiveness of various mixtures for A. bisporus disease control.     

Performance of legume–grass mixtures in the West Balkan region

Three perennial legumes (alfalfa, red clover and birdsfoot trefoil) and four cool-season perennial grasses (orchardgrass, tall fescue, Italian ryegrass and red fescue) were grown in legume–grass combinations and in pure stands of individual species, at three locations in the West Balkan region (Novi Sad, Banja Luka and Pristina) in the period from 2012 to 2015. The study evaluated dry matter yield, legume–grass–weed proportion and forage quality. High annual forage yield of legume–grass mixtures can be obtained with proper selection of species and an appropriate legume–grass ratio. However, high and stable yield, particularly in the case of grasses, depends on the amount and schedule of precipitation as well as the cutting time. The mixtures and legume pure stands achieved better forage production both per cutting and on the annual basis and had better forage quality than grass pure stands.     

Chemical contamination in upper horizon of Haplic Chernozem as a transformation factor of its physicochemical properties

Purpose

The effect of Cu, Zn, and Pb high rates on the physical properties and organic matter of Haplic Chernozem (Clayic) (A1 horizon 0–20 cm) under model experimental conditions was studied.

Materials and methods

In a model experiment, soil samples of Haplic Chernozem (Clayic) were artificially contaminated with 2000 mg/kg of Cu, Zn, and Pb acetates added separately. The particle-size fraction, the microaggregates distribution, the structural status, the total content and fractional and group composition of organic matter, physico-mechanical properties were determined in soil without metals and soil contaminated with metals.

Results and discussion

At the soil contamination with Cu, Zn, and Pb, the content of organo-mineral colloids increased, which results to the increasing of the clay fraction content by 4.5% compared to the control. The analysis of the microaggregate size composition of the studied soil shows that the content of coarser aggregates (1–0.25 mm) increases and the content of finer (0.05–0.001 mm) aggregates decreases after the addition of HMs and correspond to the HMs series: Cu → Zn → Pb. A significant decrease in the coefficient of water stability in the control from 3.0 to 1.4–1.5 in the contaminated treatments. The structural status (estimated from total agronomically valuable aggregates) changes from excellent to good. The addition of Cu, Zn, and Pb to the soil affects the quantitative composition of organic matter. The contents of free and sesquioxide-bound humic acids and free fulvic acids increased. The contamination with Zn and Pb causes the aliphatization of organic matter.

Conclusions

Under conditions of model experiment, the contamination of Haplic Chernozem (Clayic) with high rates of Cu, Zn, and Pb leads to changes of the microaggregates distribution, the structural status, and the qualitative composition of organic matter.

     

Efficient reduction of chitosan molecular weight by high-intensity ultrasound: underlying mechanism and effect of process parameters

The degradation of chitosan by high-intensity ultrasound (HIU) as affected by ultrasound parameters and solution properties was investigated by gel permeation chromatography coupled with static light scattering. The molecular weight, radius of gyration, and polydispersity of chitosan were reduced by ultrasound treatment, whereas chitosan remained in the same random coil conformation and the degree of acetylation did not change after sonication. The results demonstrate that (1) the degradation of chitosan by ultrasound is primarily driven by mechanical forces and the degradation mechanism can be described by a random scission model; (2) the degradation rate is proportional to M w (3); and (3) the degradation rate coefficient is affected by ultrasound intensity, solution temperature, polymer concentration, and ionic strength, whereas acid concentration has little effect. Additionally, the data indicate that the degradation rate coefficient is affected by the degree of acetylation of chitosan and independent of the initial molecular weight.     

Dynamics of insecticide resistance in Bemisia tabaci: a case study with the insect growth regulator pyriproxyfen

The dynamics of pyriproxyfen resistance in Bemisia tabaci (Gennadius) have been studied intensively in cotton fields and greenhouses in Israel. High resistance to pyriproxyfen evolved one year after its introduction for use on flowers in greenhouses, after three successive applications. After ten years of pyriproxyfen use in cotton fields, with only one application per season, a high level of resistance was observed, but its rate of development differed among localities. It is hypothesized that these differences reflect temporal and spatial variations in the availability of alternative host plants for B tabaci, and that pyriproxyfen resistance evolved most readily in geographically isolated areas. It is also likely that the occurrence and development of resistance is partially biotype-related. Pyriproxyfen use has ceased in high-resistance localities, but in areas with low frequencies of resistance, cotton growers have continued to achieve acceptable control of B tabaci with pyriproxyfen. Due to the absence of applications of pyriproxyfen in some cotton fields, resistance levels tended to decline between 1998 and 2001. Laboratory experiments support the hypothesis that this decline reflects, in part, fitness costs associated with pyriproxyfen resistance.