The use of sewage sludge as soil amendment. The need for an ecotoxicological evaluation

Background, aim, and scope  Sewage sludge use in agriculture should be limited by the presence of metals and other persistent environmental pollutants. The present study aims to contribute for the definition of a test battery of ecotoxicological assays that allows a proper ecotoxicological characterization of sludges, providing information on their potential hazard and identified “safe” application levels. Materials and methods  Three sludges from distinct sources (urban, olive-processing, and electroplating industries) were tested using avoidance and reproduction tests with earthworms (Eisenia andrei) and springtails (Folsomia candida) and plant growth tests with turnips (Brassica rapa) and oats (Avena sativa). Different soil–sludge mixture concentrations mimicking recommended/realistic field dosages were tested. Results  Only the sludge from the electroplating industry induced an avoidance response from the earthworms (EC₅₀ = 0.4 t/ha) and collembolans (no observed effect concentration (NOEC) = 15 t/ha). This sludge was the only sludge responsible for any effect on the reproductive output of the earthworms (EC₅₀ = 7.74 t/ha). Regarding collembolans, none of the sludges tested caused any significant decrease in reproduction. In higher plant tests, the two industrial sludges were toxic, causing a decrease growth in both species. The EC₂₀ values determined for B. rapa were 20.3 and 24.2 t/ha and for A. sativa 14.7 and 16.2 t/ha for sludges from olive-processing and electroplating industries, respectively. Discussion  The metal loadings of the different test sludges could partially explain the results obtained. The toxicity of the test sludge from electroplating industry observed on the tested invertebrates and plants could be explained by the high amount of total chromium from which 22.3% was in the most toxic oxidation state—Cr(VI). However, the toxicity caused by the sludge from the olive-processing industry in the test plants could be attributed to the presence of other compounds (not measured in this study) since the metal content was not high enough to induce such an effect. The absence of toxicity showed by the urban test sludge was in agreement with its low levels of metals. Conclusions  The response of the different test organisms and end points varied according to the sludge type. The urban sludge was non-toxic whereas the sludge from the electroplating industry caused a toxic effect on almost all parameters measured (avoidance behavior of both test organisms, reproduction of earthworms, and growth of both plant species). Sludge from the olive-processing industry only caused a toxic effect on growth of both plant species. By analyzing the sensitivity of the different parameters for the most toxic sludge, it was found that avoidance and reproduction were more sensitive than plant growth, whereas plant seed germination was not sensitive at all. Recommendations and perspectives  The ecotoxicological evaluation of wastes can be used as an environmental safety control of sludge use in agriculture. A tiered approach could be adopted for this purpose, incorporating avoidance tests in the first tier (screening level) and reproduction and plant growth tests in a second tier. But more evidence aiming to define the most suitable ecotoxicological test battery for specific sludges with a different contamination profile is still needed.     

Conserving genomic variability in large mammals: Effect of population fluctuations and variance in male reproductive success on variability in Yellowstone bison

Loss of genetic variation through genetic drift can reduce population viability. However, relatively little is known about loss of variation caused by the combination of fluctuating population size and variance in reproductive success in age structured populations. We built an individual-based computer simulation model to examine how actual culling and hunting strategies influence the effective population size (N_e) and allelic diversity in Yellowstone bison over 200 years (～28 generations). The N_e for simulated populations ranged from 746 in stable populations of size 2000 up to 1165 in fluctuating populations whose census size fluctuates between 3000 and 3500 individuals. Simulations suggested that ～93% of allelic diversity, for loci with five alleles will be maintained over 200 years if the population census size remains well above ～2000 bison (and if variance in male reproductive success is high). However for loci with 20 alleles, only 83% of allelic diversity will be maintained over 200 years. Removal of only juveniles (calves and yearlings) resulted in longer generation intervals which led to higher maintenance of allelic diversity (96%) after 200 years compared to the culling of adults (94%) when the mean census size was 3250 (for loci with five alleles). These simulations suggest that fluctuations in population census size do not necessarily accelerate the loss of genetic variation, at least for the relatively large census size and growing populations such as in Yellowstone bison. They also suggest that the conservation of high allelic diversity (>95%) at loci with many alleles (e.g., ?5) will require maintenance of a populations size greater than approximately 3250 and removal of mainly or only juveniles.     

Skin and coat traits in sheep in Brazil and their relation with heat tolerance

The objective of this study was to evaluate and compare physical parameters in groups of sheep with different phenotypic characteristics in the Centre-west region of Brazil. Five groups of sheep, with nine animals per group, were selected, three groups of Santa Inês animals with different coat colours (white, brown and black), one group with crossbred animals (Santa Inês × Bergamasca) and one group with animals of the Bergamasca breed. The following traits were evaluated: coat thickness, number and length of hair, pigmentation level in the coat and the skin as well as the percentage area of sweats glands in the skin tissue, carried out by histological analysis. The number of hairs and the area of sweats glands were not significantly different between the evaluated groups. The Bergamasca breed showed low pigmentation of the skin and long hairs. The levels of pigmentation of the hair and of the skin were highly correlated. Between the Santa Inês groups, the group with white hair showed the better parameters for heat adaptation, while the brown hair group showed the lower heat adaptation when compared with another hair breed groups.     

Effects of exogenous amylase on the in vitro digestion kinetics of whole-crop maize silages made from flint or dent grain type at different phenological stages grown in tropical condition

The effect of exogenous amylase on the in vitro rumen digestion kinetics of whole-crop maize silage made from dent (RB9004) or flint grain type (RB9308) was evaluated at different phenological stages: soft dough (SOD), early dent (EAD), ½ milkline (½M) and ¾ milkline (¾M). Forage was harvested from 70 to 110 days after sowing. Two rumen-cannulated cows receiving or not exogenous amylase (0.7 g/kg dry matter—DM, provided to achieve 396 kilo Novo units of amylase activity/kg of TMR DM) were used as donor of ruminal fluid. The in vitro gas production kinetics was evaluated according to a dual-pool logistic model. The chemical composition and gas production kinetics were affected by the hybrid and phenological stages. The flint hybrid had lower range for chemical analysis among physiological stages. Harvesting at ½M and ¾M improved DM content, bromatological composition and silage quality parameters compared to dent or flint types. Amylase (i) increased methane (CH₄) production and in vitro dry matter digestibility (IVDMD) in ½M stage, (ii) improved digestion kinetics by reducing lag time and increasing total gas production and fermentation rates of non-fibrous carbohydrates (NFC) and fibrous carbohydrates (FC), and (iii) increased extent and fermentation rate of NFC and increased fermentation rate of FC fraction in whole-crop maize silages produced from dent or flint types in all phenological stages. Harvesting between ½M and ¾M is the best phenological stage to improve chemical composition and silage quality parameters. Exogenous amylase showed improvements on fibre digestion of silages at ½M and ¾M phenological stages in both grain types of corn.     

INFLUENCE OF NITROGEN FERTILIZATION ON NICKEL ACCUMULATION AND CHEMICAL COMPOSITION OF COFFEE PLANTS DURING FRUIT DEVELOPMENT

Nutritional and physiological significance of micronutrients in coffee plants, especially with regard to nickel (Ni) is still unknown. The dynamics of nitrogen (N), phosphorus (P), potassium (K) and Ni accumulation in coffee fruits, as well as their relationships with total soluble protein, amino acids, reducing sugars, and starch content during coffee fruit development (green, ripe, and dry fruits), were investigated. Coffee trees received three N fertilizer rates (0, 150, and 300 kg of N ha^?1) as ammonium sulfate split into three applications per year. Nitrogen fertilization increased reducing sugars and starch concentrations in ripe fruits. In contrast, green fruits showed the highest amino acid and Ni concentrations. Fruit Ni concentration decreased in both green and ripe fruits as N rates increased; thus, indicating the possibility of either a N-associated dilution effect on Ni concentration or that Ni uptake by roots and/or transport to developing fruit was limiting. Plant nutritional status and fruit development stage influenced the coffee grain chemical composition. Furthermore, the variation in reducing sugars and starch content was more closely linked to the stage of fruit development than to N supply. A supposed relationship among the decreased of caffeine, starch, amino acids, and proteins with Ni content during green fruit development suggests a fundamental role for Ni in coffee fruit ripening. The interaction between N and Ni metabolism during fruit ripening might influence the chemical parameters involved in the coffee grain quality. This is the first report documenting changes in Ni concentrations of coffee fruit as a function of N fertilization rates and the development stage, but further research is needed to better understand the significance of N-Ni interaction in developing coffee fruit.     

Encapsulated nitrate replacing soybean meal changes in vitro ruminal fermentation and methane production in diets differing in concentrate to forage ratio

The objective of this study was to evaluate the effects of using encapsulated nitrate product (ENP) replacing soybean meal in diets differing in concentrate to forage ratio on ruminal fermentation and methane production in vitro using a semi‐automatic gas production technique. Eight treatments were used in a randomized complete design with a 2 × 4 factorial arrangement: two diet (20C:80F and 80C:20F concentrate to forage ratio) and four levels of ENP addition (0%, 1.5%, 3.0%, and 4.5% of DM) replacing soybean meal. There was a diet × ENP interaction (p = 0.02) for methane production. According to ENP addition, diets with 80C:20F showed more intense reduction on methane production that 20C:80F. A negative linear effect was observed for propionate production with ENP addition in diet with 80C:20F and to the relative abundance of methanogens Archaea, in both diet. The replacement of soybean meal by ENP in levels up to 3% of DM inhibited methane production due to a reduction in the methanogens community without affecting the organic matter degradability. However, ENP at 4.5% of DM level affected fiber degradability, abundance of cellulolytic bacteria, and propionic acid production, indicating that this level of inclusion is not recommended for ruminant production.     

Effect of red propolis extract isolated or encapsulated in nanoparticles on the in vitro culture of sheep preantral follicle: Impacts on antrum formation,mitochondrial activity and glutathione levels

This study aimed to characterize the hydroethanolic extract of red propolis (HERP) and nanoparticles containing HERP for using as an additive in the culture medium of isolated ovine preantral follicles. HERP was characterized by high‐performance liquid chromatography (HPLC) and determination of flavonoid content, and the nanoparticles by the mean particle diameter, polydispersity index (PI) and encapsulation efficiency (EE). The effect of HERP (10 and 20 ηg/ml—HERP10 and HERP20 groups) and nanoparticles (NP10 and NP20 groups) on isolated secondary follicles cultured in vitro for 12 days was observed by morphological evaluation, oxidative stress markers (reactive oxygen species—ROS and glutathione—GSH) and active mitochondria. HPLC showed formononetin as the major compound in the HERP (63.92 ± 0.21 μg/mL). The content of flavonoids ranged from 2.14% to 11.00%. The nanoparticles showed mean diameter of 287.5 ± 3.9 and 479 ± 18.1 ηm; PI of 0.117 ± 0.018 and 0.316 ± 0.039; and EE of 67.15% and 41%, respectively, for the NP10 and NP20 groups. After 12 days of culture, HERP20 and NP20 increased (p < 0.05) the percentage of normal follicles compared to NP10. HERP20 showed significantly higher percentages of antrum formation than control medium (MEM) and NP10 (p < 0.05). HERP20 also showed an increase (p < 0.05) in mitochondrial activity compared to the other treatments, except NP20 (p > 0.05), and increased GSH levels (p < 0.05) compared to MEM and HERP10. In conclusion, use of HERP (20 ηg/ml) on in vitro culture of isolated ovine preantral follicles can increase antrum formation, mitochondrial activity and GSH levels.