Increasing level of liquid pig manure reduces Eisenia andrei and Enchytraeus crypticus reproduction in subtropical soils.

Wastes generated in pig production are widely used as agricultural fertilizers. Nevertheless, such form of disposal supplies large amounts of waste in soils annually and can cause environmental pollution. The ecological risk of this practice to soil organisms has received little attention. Ecotoxicological tests are used to evaluate the toxicity of contaminants added to the soil biota. The aimed to evaluate the effect of liquid pig manure (LPM) on the reproduction of Eisenia andrei and Enchytraeus crypticus when applied in natural soils. LPM doses caused effects on earthworm reproduction in both soils, with EC50 of 112 and 150 m3 ha-1 in the Entisol and Nitosol, respectively. On enchytraeids, LPM had bigger effects, leading to EC50 of 17.7 and 45.0 m3 ha-1 in the Entisol and Nitosol, respectively. The results emphasize the importance to consider the ecological risks of LPM of conducting studies with natural soils and edaphic fauna as indicators.

Different diets with and without inclusion of antimicrobial additives alter the toxicity of swine manure to springtails and earthworms? / Diferentes dietas com e sem inclusão de aditivos antimicrobianos alteram a toxicidade do esterco de suíno para colêmbolos e minhocas?

The objective of this study was to measure the impact on part of soil fauna of application of non-stabilized (fresh) manure from post-weaning pigs fed diets, formulated with or without the use of dual-purpose wheat (15% inclusion), and with or without the use of antimicrobial growth-promoting additives (100 mg kg-1 doxycycline + 50 mg kg-1 colistin + 250 mg kg-1 Zn oxide). Two species of edaphic organisms were evaluated, the springtails Folsomia candida and the earthworms Eisenia andrei, using ecotoxicological avoidance behavior tests. The treatments were swine manure from: RR: Reference Ration; WR: Wheat Reference; RA: Reference Ration + Antimicrobial Additives; WA: Wheat Reference + Antimicrobial Additives. The doses of waste used for treatments were as follows: 0 (control), 5, 10, 20, 30, 40, 65 and 100 m³ ha-1 for the springtails, and 0 (control), 5, 10, 20, 30, 40 and 65 m³ ha-1 for the earthworms, applied in the Oxisol. The experimental design was completely randomized with four replicates. The use of non-stabilized swine manure did not affect the avoidance behavior of F. candida at any dose, regardless of the use of antimicrobial additives or wheat. For E. andrei, there was avoidance behavior at all treatments and doses used. These avoidance behaviors were related to the sensitivity of each species of soil organism. The avoidance behavior for earthworms was related to the doses of non-stabilized swine manure in soil and not to the various diets and/or the use of growth-promoting additives

O estudo foi realizado com objetivo de mensurar o impacto sobre parte da fauna do solo de aplicações de dejetos sem estabilização (frescos), provenientes de suínos na fase de creche arraçoados com diferentes dietas, formuladas com ou sem o uso do trigo de duplo propósito (15% de inclusão), e com ou sem o uso de aditivos antimicrobianos promotores de crescimento (100 mg kg-1 de doxiciclina + 50 mg kg-1de colistina + 250 mg kg-1 de óxido de Zn). Foram avaliadas duas espécies de organismos edáficos, colêmbolos Folsomia candida e minhocas Eisenia andrei, com o uso de ensaios ecotoxicológicos de comportamento de fuga. Os tratamentos foram dejetos de suínos provenientes de: RR: Ração Referência; WR: Trigo Referência; RA: Ração Referência + Aditivos Antimicrobianos; WA: Trigo Referência + Aditivos Antimicrobianos. As doses de dejetos utilizadas para os tratamentos foram: 0 (controle), 5, 10, 20, 30, 40, 65 e 100 m³ ha-1 para os colêmbolos, e 0 (controle), 5, 10, 20, 30, 40 e 65 m³ ha-1 para as minhocas, aplicadas em Latossolo Vermelho distrófico. O delineamento experimental foi inteiramente casualizado com quatro réplicas. A utilização de dejetos de suínos não estabilizados não afetou a fuga de F. candida em nenhuma das doses, independente do uso ou não de aditivos antimicrobianos ou de trigo. Já para E. andrei o comportamento foi inverso e houve fuga em todos os tratamentos e doses utilizados. Os comportamentos de fuga foram relacionados à sensibilidade de cada espécie de organismo edáfico. O comportamento de fuga para minhocas foi relacionado com as doses dos dejetos de suínos não estabilizados no solo e não às várias dietas e/ou ao uso dos aditivos promotores de crescimento

Impacts on reproduction of Enchytraeus crypticus in fertilized soils with chicken litter treated with synthetic and natural insecticide.

Poultry litter is used as a fertilizer due to the high concentration of nutrients. However, this material receives application of medicines, whether they are used to treat animals or the litter itself. Thus, the objective was to evaluate if the application in soils of poultry litter doses treated with two insecticides (cypermethrin and cinnamon oil) used to control of the Alphitobius diaperinus, affect the reproduction of Enchytraeus crypticus, and the insecticides effects over time, trough of ecotoxicological tests using soils with different textural classes an Entisol and an Oxisol. In the Oxisol the application of poultry litter stimulated the E. crypticus reproduction regardless of the insecticide and the collection time. In Entisol the poultry litter showed a toxicity at 10 t ha-1 when treated with cypermethrin and at 30 t ha-1 with cinnamon oil. Entisol results show that cypermethrin affects non-target edaphic organisms soon after spraying, but over time has reduced toxicity unlike cinnamon oil.

Eprinomectin antiparasitic affects survival, reproduction and behavior of Folsomia candida biomarker, and its toxicity depends on the type of soil.

The objective of this study was to evaluate the toxicity of the antiparasitic agent eprinomectin in two subtropical soils, using ecotoxicological lethality, reproduction and avoidance behavior tests with springtails (Folsomia candida). Eprinomectin concentrations were 0 (control), 0.5, 1, 2, 4, 8, 12, 16 and 20 mg kg-1 of dry soil combined with either Entisol or Oxisol soils. Statistically significant toxic effects of eprinomectin on springtails were observed in both soils. Eprinomectin was lethal starting at 8 mg kg-1 of dry soil in Entisol, and 20 mg kg-1 of dry soil in Oxisol, with effects less than 50% at lethal concentrations. Reductions in the reproduction rate of the springtails were also observed starting at 8 mg kg-1 of dry soil in Entisol, and 0.5 mg kg-1 of dry soil in Oxisol. ECrepr50 value calculated for Entisol was 4.38 ±â€¯0.62 mg kg-1 of dry soil; for Oxisol the ECrepr50 was above the highest tested concentration. For avoidance behavior, the effect occurred from 0.5 mg kg-1 of dry soil for both soils. In Entisol, all concentrations caused avoidance of more than 95%, and in Oxisol the ECavoi50 value was 1.33 ±â€¯0.83 mg kg-1 of dry soil. We conclude that eprinomectin affected survival, reproduction and caused avoidance behavior of F. candida in both soils. The toxic effects were greater as the concentration in the soils increased. The effects in Oxisol were less intense than those in Entisol with respect to the affected springtails. This discrepancy may be attributed to the different physicochemical characteristics of the soils that determine the retention capacity for eprinomectin; in particular, there are greater contents of clay, organic matter and cation exchange capacity in Oxisol.

Fish exposed to water contaminated with eprinomectin show inhibition of the activities of AChE and Na+/K+-ATPase in the brain, and changes in natural behavior.

The objective of this study was to evaluate whether antiparasitic eprinomectin may be an environmental contaminant in water compartment in low concentrations, negatively affecting neurotransmission and, consequently, the natural behavior of the jundiá (Rhamdia quelen). Fish were randomly allocated in tanks and exposed for 24 and 48 h to eprinomectin concentrations in water [0.0 (Control), 1.124 (T1), 1.809 (T2) and 3.976 (T3) µg L-1], followed by 48 h of recovery in eprinomectin-free water, in order to evaluate the behavioral parameters, levels of reactive oxygen species (ROS) in the brain, as well as cerebral enzymatic activities of acetylcholinesterase (AChE) and of the sodium-potassium ATPase pump (Na+/K+-ATPase). Especially at the two highest concentrations of eprinomectin (T2 and T3), the fish showed alterations in natural behavior, particularly hyperlocomotion and longer time on the surface. Furthermore, at these same concentrations, cerebral ROS levels increased and cerebral AChE activity decreased. At the highest concentration (T3) cerebral Na+/K+-ATPase activity was reduced. Increased ROS and impairment of AChE and Na+/K+-ATPase enzymes in the brain may have contributed directly to behavioral changes, due to neuronal damage and synapse impairment. Even after 48 h in water without eprinomectin, behavioral changes and neurotoxic effects were observed in fish, suggesting residual effects of the antiparasitic. In conclusion, eprinomectin even in low concentrations may be a hazardous environmental contaminant for aquatic organisms, as it causes brain damage and affects the natural behavior of fish.

Ecotoxicological assessment of Fluazuron: effects on Folsomia candida and Eisenia andrei.

The cattle production in Brazil has increased considerably in the last years, mainly due to the control of parasite infestation of the animals, which cause loss of productivity to the sector. Fluazuron is an active ingredient (a.i.) of the benzoylurea class used to control ticks in cattle. As this a.i. has been found unchanged in animal feces, which may present a risk to edaphic organisms, this study aimed to assess the effects of fluazuron on survival, reproduction, and behavior of the soil invertebrates Folsomia candida and Eisenia andrei, through ecotoxicological assays. We carried out bioassays in a tropical artificial soil (TAS) spiked with increasing doses of the insecticide. Earthworm mortality was found only at the highest tested fluazuron concentration (LOEC = 160 mg a.i. kg-1 dry soil and NOEC = 80 mg kg-1), while the reproduction of F. candida and E. andrei was reduced at lower fluazuron concentrations (EC50 = 4.48 mg kg-1 and EC50 = 20.8 mg kg-1, respectively). Avoidance behavior was detected for both species at lower concentrations than those that caused impacts on reproduction, indicating that the substance may affect the soil habitat function. Since the possible adverse effects of fluazuron on edaphic fauna are still unknown or neglected, this study also warns about the possible harmful effect of veterinary pharmaceutical products on edaphic fauna.

Evaluation of cytotoxicity, genotoxicity and ecotoxicity of nanoemulsions containing Mancozeb and Eugenol.

Mancozeb is a fungicide widely used in agriculture, mostly against the pathogen Glomerella cingulata responsible for the rot of ripe grape, but presents high toxicity. Strategies are sought to reduce the toxicity of this fungicide and alternative treatments are welcome. An alternative could be the use of clove oil, which has Eugenol as its major compound, and has antifungal potential against G. cingulata, however, Eugenol is susceptible to degradation processes which may compromise its efficacy. The nanoencapsulation of Mancozeb and Eugenol is a possible strategy to overcome the limitations of toxicity, solubility and instability of these compounds. Therefore, the objective of this study is to develop nanoemulsions containing Mancozeb (0.1 mg/mL) and Eugenol (33 mg/mL), isolated or associated, and evaluate the safety of these formulations through cytotoxicity, genotoxicity and ecotoxicity tests. Nanoemulsions were developed by the spontaneous emulsification method, cytotoxicity and genotoxicity were evaluated in healthy human cells through MTT, Dichlorofluorescein diacetate and Picogreen tests, and ecotoxicity assessment was carried out using the chronic toxicity test in springtails. After preparation, the physicochemical characterization of the nanoemulsions were performed which presented mean particle size between 200 and 300 nm, polydispersity index less than 0.3, negative zeta potential and acid pH. The nanoencapsulation was able to avoid the reduction of the cell viability caused by Mancozeb, while Eugenol was shown to be safe for cell use in both free and nanostructured forms, however the association of the two active compounds showed toxicity in the higher doses of Mancozeb. In the ecotoxicity tests, both free Mancozeb and Eugenol forms presented high toxic potential for soil, whereas the nanoencapsulation of these compounds did not cause a reduction in number of springtails. Therefore, from the tests performed, it was possible to observe that nanoencapsulation of Mancozeb and Eugenol is a safe alternative for the application of these compounds mainly in agriculture.

Disentangling the effects of the aqueous matrix on the potential toxicity of liquid pig manure in sub-tropical soils under semi-field conditions.

Inadequate application of liquid pig manure (LPM) may pose risks to the soil due to the potential contaminants that exists, as well as by the large water input that can originate excessive moisture. By using Terrestrial Model Ecosystems, this study aimed to evaluate the effect of application of LPM (82% moisture) using the application rates of 20, 50 and 150 m3 ha-1 and also of the corresponding amount of water to understand the origin of effects on the soil fauna of two sub-tropical soils. In general, the results obtained for the two soil types indicated that LPM (150 m3 ha-1) changed the composition of soil fauna, with an increase in the abundance of insect larvae and dipterans, but a decrease in the number of earthworms and enchytraeids. Microbial biomass, soil respiration and the nutrients Mg, K and P increased with the amount of LPM supplied to the soil. When analysing the effects of adding large volumes of water via the LPM, results showed that application did not originate significant effects on the parameters measured. The differences obtained when comparing both treatments were mainly attributed to the organic load brought by the LPM and not to the amount of water without LPM.

Ecotoxicological effects of fipronil, neem cake and neem extract in edaphic organisms from tropical soil.

Veterinary medicines are widely applied for the treatment and prevention of animal diseases. Consequently, animal manure contains significant amounts of environmental pollutants that are potential sources of environmental pollution when inappropriately applied in soils. This work aimed to evaluate ecotoxicological effects of doses of commercial fipronil, neem cake and neem extract in the survival and reproduction of earthworms (Eisenia andrei), enchytraeidae (Enchytraeus crypticus) and springtails (Folsomia candida) in Oxisol and tropical artificial soil (TAS). Applications of fipronil, neem cake and extract in soil were carried out according to standardized ISO methodologies by using a random experimental design with five replicates. Toxic effects of fipronil for springtails in Oxisol and TAS were observed with LC50 of 0.26 mg kg-1 (0.18-0.35 mg kg-1) and 0.29 mg kg-1 (0.22-0.37 mg kg-1), respectively. It was not observed significant toxic effects of fipronil for earthworms and enchytraeidae in both soils. However, significant amounts of juvenile earthworm and adult enchytraeidae decreased in fipronil doses higher than 10 mg kg-1. Neem cake and extract were not toxic for earthworms and enchytraeidae but, significant amounts of juvenile springtails decreased in neem cake doses from 500 to 1000 mg kg-1. It can be concluded that the use of veterinary medicines containing synthetic compounds for preventing diseases in animals needs to be controlled to avoid environmental pollution after applying manure in soil. Veterinary medicines containing natural compounds as neem cake and extract are eco-friendly and could be efficiently applied in soil in a sustainable way.

Ecotoxicological evaluation of swine manure disposal on tropical soils in Brazil.

Swine production in Brazil results in a great volume of manure that normally is disposed of as agricultural fertilizer. However, this form of soil disposal, generally on small farms, causes the accumulation of large amounts of manure and this results in contaminated soil and water tables. To evaluate the effects of increasing concentrations of swine manure on earthworms, several ecotoxicological tests were performed using Eisenia andrei as test organism in different tropical soils, classified respectively as Ultisol, Oxisol, and Entisol, as well as Tropical Artificial Soil (TAS). The survival, reproduction and behavior of the earthworms were evaluated in experiments using a completely randomized design, with five replications. In the Ultisol, Oxisol and TAS the swine manure showed no lethality, but in the Entisol it caused earthworm mortality (LOEC=45 m(3)ha(-1)). In the Entisol, the waste reduced the reproductive rate and caused avoidance behavior in E. andrei (LOEC=30 m(3)ha(-1)) even in lower concentrations. The Entisol is extremely sandy, with low cation exchange capacity (CEC), and this may be the reason for the higher toxicity on soil fauna, with the soil not being able to hold large amounts of pollutants (e.g. toxic metals), but leaving them in bioavailable forms. These results should be a warning of the necessity to consider soil parameters (e.g. texture and CEC) when evaluating soil contamination by means of ecotoxicological assays, as there still are no standards for natural soils in tropical regions. E. andrei earthworms act as indicators for a soil to support disposal of swine manure without generating harm to agriculture and ecosystems.