Phosphorus removal from urban wastewater through adsorption using biogenic calcium carbonate.

Phosphorus (P) removal from urban wastewater is increasingly relevant in the wastewater treatment sector. The present work aims to contribute to the study of the adsorption process as a P removal technology. Biogenic calcium carbonate from industrial eggshell waste prepared by milling and calcination was used as an adsorbent. Batch adsorption experiments were conducted using real wastewater with 40 mg P/L (orthophosphate), original pH 7.33, under stirring conditions (100 rpm). The adsorbent was characterized using SEM-EDS, XRD, and FTIR-ATR before and after adsorption. From an initial screening of calcination times (15, 30, 60, and 120 min) and considering a balance between P removal and energy saving, the adsorbent selected was eggshell calcined at 700 °C for 60 min. The Langmuir isotherms describe the experimental data with a maximum adsorption capacity of 4.57 mg P/g at 25 °C. The adsorption process reached equilibrium within 120 min for different dosages (5, 10, and 20 g/L at 25 °C). Batch experiments showed that SO42-, at a concentration of 2689 mg/L reduced the P adsorption selectivity for dosages ≤10 g/L at 25 °C. Characterization of the loaded adsorbent shows that P adsorption from real wastewater is mostly electrostatic attraction, with the contribution of ligand exchange and microprecipitation. The adsorption capacity and behavior of the selected adsorbent seem promising for P removal from urban wastewater compared with other low-cost adsorbents.

Effects of fresh and field-aged holm-oak biochar on As, Cd and Pb bioaccumulation in different rice growing environments.

Arsenic, Cd, and Pb environmental fate is influenced when the traditional permanent flooding rice production systems are replaced by water-saving and soil conservation practices, urging for additional strategies that avoid their bioaccumulation in rice grain. The aim of this two-years field study was to evaluate the effects of fresh and field-aged biochar on As, Cd, and Pb bioaccumulation, and on As speciation, in rice grain produced in different growing environments (flooding versus sprinkler and conventional tillage versus direct seeding). Biochar produced from holm-oak pruning residues (pyrolysis at 550 °C, 48 h), in a single application (28 Mg ha-1), reduced As bioaccumulation in rice grain in the permanent flooding system to non-quantifiable concentrations (e.g., from 0.178 mg kg-1 to <0.04 mg kg-1, for inorganic-As, respectively), an effect which remained under field-aging conditions, increasing rice commercial value. When adopting sprinkler irrigation, the undesirable increase in Cd bioaccumulation in rice, relatively to the anaerobic system, was counteracted by biochar application, reducing its bioaccumulation in kernels between 32 and 80 %, allowing a simultaneous control of metals and metalloids bioaccumulation in rice. The bioaccumulation of Pb was also prevented with biochar application, with a reduction in its concentration four- to 13-times, in all the management systems, relatively to the non-amended plots, under fresh biochar effects. However, Pb immobilization decreased with biochar field-aging, indicating that the biochar application may have to be repeated to maintain the same beneficial effect. Therefore, the present study shows that the implementation of sprinkler irrigation with holm-oak biochar could reduce the risk of heavy metals(loids) bioaccumulation in rice grains and, thereby, ensuring food safety aspects, particularly under fresh biochar effects.

Integrative toolbox to assess the quality of freshwater sediments contaminated with potentially toxic metals.

The Guadiana Basin is a transnational basin, presenting historical contamination with potentially toxic metals (PTM), which origin can be both natural and anthropogenic. This study explores the use of a set of observational, chemical and ecotoxicological assays with Heterocypris incongruens, Vibrio fischeri, Pseudokirchneriella subcapitata, Thamnocephalus platyurus, identifying the most sensitive to be included in a toolbox to analyze the quality of freshwater sediments related to this type of contamination. The study included the analysis of a reservoir and streams sediments of Guadiana basin, in two consecutive years with different climate conditions 2017 (dry year) and 2018 (normal year). The results showed high chemical variability along the basin, with greater contamination with PTM in the reservoir sediments. The calculated Enrichment Factors (EF) indicated high anthropogenic contamination by Cd, followed by Pb (EF > 1.5). The geoaccumulation index (Igeo) revealed that the sediments were severely polluted with Cd, and slightly polluted with Pb and Cu, inducing a higher sublethal toxicity to Heterocypris incongruens. Among the parameters evaluated, and after the use of multivariate statistical techniques, the toolbox for assessing sediments quality, in similar climate and geological conditions, should include the analysis of: meteorology, land use/cover in the area, granulometry, organic matter content, PTM concentrations, contamination indices (e.g., Igeo and EF), and sublethal bioassays with H. incongruens (total sediment analysis) and Vibrio fisheri luminescence inhibition (pore water analysis).

Evaluation of the fertilizer potential of Chlorella vulgaris and Scenedesmus obliquus grown in agricultural drainage water from maize fields.

Producing microalgae with agricultural drainage water (ADW) allows recycling water and nutrients, with the production of a biofertilizer, avoiding receiving waters' contamination. Chlorella vulgaris and Scenedesmus obliquus were cultivated using ADW and standard media supplementation and presented higher productivities, relatively to the control industrial growth medium (using freshwater). Selected strains were grown outdoors in pilot flat panel photobioreactors, reaching 2.20 g L-1 for S. obliquus and 1.15 g L-1 for C. vulgaris, and degrading herbicides in the ADW to non-quantifiable concentrations. The potential of the C. vulgaris and S. obliquus suspensions to replace 50% of nitrogen (N) mineral fertilization of lettuce (0.5 g pot-1) was evaluated through a pot trial, also using a 2-times (1.0 g pot-1) and 5-times (2.5 g pot-1) higher dose, applied 31 days before lettuce transplanting. Even the lower dose of N, applied via C. vulgaris or S. obliquus suspensions, was able to provide significantly higher lettuce fresh matter yield, relatively to the mineral fertilized control. Soil enzymatic activities were improved, with significantly higher dehydrogenase, ß-glucosidase, and acid phosphatase activities for the 2.5 g pot-1 dose, more marked for S. obliquus, which was also able to increase soil organic matter content. Both the non-fertilized control and microalgae fertilized pots led to similar soil electrical conductivities, 3-fold lower than in the N-mineral fertilized pots, evidencing the capacity of microalgae fertilizers to avoid soil secondary salinization. Results suggest benefits from using ADW from maize cultivation to produce C. vulgaris or S. obliquus suspensions, that can be further used as liquid organic slow-release fertilizer.

Characterization of preantral follicle clustering and neighborhood patterns in the equine ovary.

Understanding the transition from quiescent primordial follicles to activated primary follicles is vital for characterizing ovarian folliculogenesis and improving assisted reproductive techniques. To date, no study has investigated preantral follicle crowding in the ovaries of livestock or characterized these crowds according to follicular morphology and ovarian location (portions and regions) in any species. Therefore, the present study aimed to assess the crowding (clustering and neighborhood) patterns of preantral follicles in the equine ovary according to mare age, follicular morphology and developmental stage, and spatial location in the ovary. Ovaries from mares (n = 8) were collected at an abattoir and processed histologically for evaluation of follicular clustering using the Morisita Index and follicular neighborhoods in ovarian sections. Young mares were found to have a large number of preantral follicles with neighbors (n = 2,626), while old mares had a small number (n = 305). Moreover, young mares had a higher number of neighbors per follicle (2.6 ± 0.0) than old mares (1.2 ± 0.1). Follicle clustering was shown to be present in all areas of the ovary, with young mares having more clustering overall than old mares and a tendency for higher clustering in the ventral region when ages were combined. Furthermore, follicles with neighbors were more likely to be morphologically normal (76.5 ± 6.5%) than abnormal (23.5 ± 6.5%). Additionally, morphologically normal activated follicles had increased odds of having neighbors than normal resting follicles, and these normal activated follicles had more neighbors (2.6 ± 0.1) than normal resting follicles (2.3 ± 0.1 neighbors). In the present study, it was demonstrated that preantral follicles do crowd in the mare ovary and that clustering/neighborhood patterns are dynamic and differ depending on mare age, follicular morphology, and follicular developmental stage.

Effect of retained placenta and clinical mastitis on reproduction parameters, immune response, and steroidogenic receptors gene expression in postpartum crossbred dairy cows.

The purpose of this study was to evaluate the effect of retained placenta (RP) and clinical mastitis (CM) on the reproductive efficiency of crossbred dairy cows during the postpartum period and the effect in some innate immune system indicators. For this, two experiments were carried out. In the first, a total of 232 cows were evaluated and divided as: healthy control (n = 184), RP (n = 22), and CM (n = 26) groups. The RP and CM was evaluated until 30 days postpartum (DPP) and reproductive rates were measured. In experiment 2, cows were divided in control (n = 10), RP (n = 10), and CM (n = 30) groups. Between 40 and 50 DPP, clinical, gynecological examination and endometrial cytobrush were performed to evaluate subclinical endometritis (SE) and gene expression of interleukins 1ß (IL-1ß) and 6 (IL-6), chemokine ligand 5 (CCL5), estrogen α (ESR1), and progesterone (PGR) receptors by qRT-PCR analysis. In experiment 1, the conception rate at 1st artificial insemination (AI) was lower in RP and CM groups and pregnancy rate at 150 days decreased in CM group. Calving-to-1st AI interval and days open were shorter in healthy cows. In experiment 2, the occurrence of SE was 26.7% and higher in RP and CM groups. The expression of IL-1ß increased in RP and CM groups, while IL-6 was less expressed in RP group. The CCL5, ESR1, and PGR were similar between groups. In conclusion, cows with RP and CM had their reproductive efficiency negatively affected and had they initial pro-inflammatory response improved by the increase of IL-ß.

Combined use of olive mill waste compost and sprinkler irrigation to decrease the risk of As and Cd accumulation in rice grain.

Sprinkler irrigation has been successfully introduced in rice production as an alternative to the traditional flooding system, allowing water savings and the reduction of As accumulation in the grain. However, the same conditions can increase Cd mobility and grain accumulation, an effect that needs to be countered. A 3-year field experiment was set-up in a Mediterranean region (Extremadura, Spain), to evaluate how the application of compost from olive mill waste (single application, 80 t ha-1), influences the accumulation of As and Cd in the grain under different irrigation regimes. Accumulation of As in the grain was always lower in the sprinkler irrigation when compared with the flooding irrigation, reaching a 5-fold difference in the third year. Compost application did not evidence a clear effect on the As accumulation in the rice grain, but highly significant negative correlations (p < 0.001) were obtained between As content in the grain (total, inorganic, and organic) and the humification parameters in the soil, evidencing the importance of using a mature and stable organic amendment to avoid As accumulation in the grain. Cadmium accumulation in the rice grain decreased in each treatment where compost was applied, relatively to the non-treated counterpart (e.g., from 0.080 to <0.010 mg kg-1, in direct seeding with sprinkler irrigation, in the third year). There were no significant differences in the total inorganic As between treatments with or without compost application, but it was possible to observe an increase in the predominance of the organic As over the more toxic inorganic As, when compost was applied, allowing a decrease in the risk associated to As accumulation. Therefore, the aerobic cultivation of rice, with the simultaneous application of an adequate source of organic matter, can be considered a good solution to cope with the risk of accumulation of As and Cd in the rice grain.

Early COVID-related Acute Kidney Injury Recovery May Course with Hydroelectrolytic Disorders in Patients With High Risk of Insensible Fluid Loss

BackgroundAcute Kidney Injury (AKI) may occur in more than 30% of COVID hospitalized patients, and renal recovery is poorly described. AimWe aimed to evaluate the renal short-term recovery profile of COVID-related AKI (COV+) compared to COVID-unrelated AKI (COV-). Designcase-control retrospective single-center study MethodsAll patients admitted to the Hospital das Clinicas, University of Sao Paulo, who recovered AKI from April to June of 2020 (COV+, n=98) and from August to October of 2019 (COV-, n=50) were analyzed. Recovery was defined by spontaneous serum creatinine drop or withdrawal of dialysis. Serum electrolytes were analyzed during the first five days of recovery. ResultsAmong 333 COV+ patients, 98 recovered from AKI (29.4%), while 50 of 177 COV-patients recovered (28.2%). The COV- group presented higher prevalence of chronic morbidities, while the COV+ group had a worse acute clinical course requiring vasoactive drugs (VAD), mechanical ventilation (MV) and dialysis. COVID-19 diagnosis was associated with need of mecxhanical ventilation, dialysis, presence of fever, and higher use of any diuretic drug during first days of recovery. The presence of fever and mechanical ventilation were the predictors associated with intravascular volume depletion surrogates (daily progressive rising in sodium levels and elevation in serum urea: creatinine ratio). Neither COVID-19 nor diuretics use seem to be independent risk factors for this. ConclusionsIntravascular volume depletion surrogates are more common in short-term AKI-recovery of patients presenting fever and mechanical ventilation, commons features in SARS-CoV2 infection.

From wastewater to fertilizer products: Alternative paths to mitigate phosphorus demand in European countries.

Phosphorus (P) is a non-renewable resource, irreplaceable for life and food production, and currently considered a Critical Raw Material to the European Union (EU). Due to concerns about the rate of consumption and limited reserves in countries with sensitive geopolitical contexts, it is urgent to recover P from urban and industrial flows. Indeed, the municipal wastewater treatment plants (WWTP) are considered relevant sources with several hot spots, especially sewage sludge with estimated recovery efficiencies of >80%. The most promising recovery strategies are based on thermal treatments (e.g., incineration of sludge) following by wet-chemical or thermo-chemical leaching, precipitation, and adsorption. The direct application of sludge on soil is no longer a primary route for P reintegration in the value-chain for countries as Switzerland, Germany, and The Netherlands. In fact, Switzerland and Austria paved the way for implementing P recovery legislation, focusing on recovery from raw sewage sludge or ashes. Indeed, industrial technologies with sludge ash as input show high recovery efficiencies (Ashdec® and Leachphos® with 98 and 79%) and lower environmental impacts, whereas Pearl® technology has about 12% recovery efficiency with wastewater as input. After all, struvite emerges as the most recovered product with recent access to the internal market of EU fertilisers and similar growth performance compared to triple-super-phosphate. However, several studies leave open the possibility of introducing loaded adsorbents with P as soil amendments as a new alternative to conventional desorption. Briefly, P recovery should be a compromise between efficiency, environmental impacts, and economic revenues from the final products.

The dairy sector in the Azores Islands: possibilities and main constraints towards increased added value.

The Azores archipelago is the most suitable region for dairy production in Portugal, representing 30% of the overall Portuguese dairy production. It has a production system characterized by an average milk yield of 6216 kg/cow/year, and the predominance of pasture-based feeding and cows that have longer productive lives and lower incidence of metabolic/production diseases, such as acidosis or mastitis. The biggest problem with the Azores Islands dairy sector is the cost of transport, as the main markets are located in continental Portugal, over 1500 km away, and local dairy products have to compete with dairy products produced in mainland Portugal and in the rest of the European Union. Herein, the evolution of the dairy sector in the Azores Islands from 2007 to 2017 is presented. A SWOT (Strength, Weakness, Opportunity, and Threat) analysis was performed to find potential solutions to increase the value of the Azorean dairy sector. The most relevant solution considered was the valorization of the dairy production through three major aspects: higher milk quality, namely, better organoleptic properties; lower carbon footprint (not considering transport costs); and higher levels of animal welfare. Three examples are shown of such valorization: protected denomination of origin (PDO) cheeses, the "happy cows" program, and the production of an organic milk, from the Terceira Island. Some of these programs are relatively recent, so, it will be interesting to see how their sales and acceptance by consumers evolve, particularly under the current economic framework.

Leptospirosis in slaughtered cows in the Triangulo Mineiro, Minas Gerais: prevalence, serological profile and renal lesions / Leptospirose em vacas abatidas no Triângulo Mineiro, Minas Gerais: prevalência, perfil sorológico e lesões renais

Leptospirosis is a zoonosis that affects several species of domestic and wild animals and is an important cause of economic losses in cattle in Brazil. In this study, we determined the prevalence of bovine leptospirosis in the Triângulo Mineiro region, Minas Gerais, Brazil, identified the most frequent serovars of Leptospira interrogans, and examine the renal pathological changes associated with the disease. Samples of blood serum and kidney fragments from 100 bovine females were collected in cattle abattoir. In the serological investigation 48% of the cows were positive. The serovars for which there were more reactive animals were Wolffi (24%), Hardjo (21%) and Hebdomadis (18%). Among the positive samples, 14/48 showedantibody titers greater than 1:100, and 70.83% of the seropositive animals responded to more than one Leptospira interrogans serovar. Only one farm did not have seropositive cows and in nine farms studied, six (66.66%) presented seropositive animals to the Hebdomadis serovar. At the histopathological examination, the most frequent microscopic lesions in positive animals were hyalinization (81.25%), congestion (81.25%) and hydropic degeneration (70.83%). However, these histopathological alterations were also found in kidneys of animals negative to serology, such as hyalinization (80.77%), congestion (48.07%) and hydropic degeneration (55.77%) and these findings are unrelated to positivity. Histopathological examination of the kidneys is not indicated to replace the serological diagnosis of leptospirosis, and may be used only as a complementary examination. Despite the low frequency of seropositive animals in the Triângulo Mineiro region, the disease is present in a large number of farms. Noteworthy is the high frequency of serovar Hebdomadis and it can be considered an emerging serovar in the region. The evaluation of the frequency of this serovar in other regions becomes important, and once verified should result in the recommendation of the inclusion of this serovar in the leptospirosis control.

A leptospirose é uma zoonose que afeta várias espécies de animais domésticos e silvestres e importante causa de prejuízos à bovinocultura nacional. Objetivou-se determinar a prevalência da leptospirose e identificar os sorovares de Leptospira interrogans mais freqüentes na região do Triângulo Mineiro, além de avaliar microscopicamente as lesões renais e correlacionar estas lesões com sorovares específicos. Foram coletadas amostras de soro sanguíneo e fragmentos de rim de 100 fêmeas bovinas em abatedouro e, destas 100 amostras, 48% foram positivas. Os sorovares para os quais houve mais animais reagentes foram Wolffi (24%), hardjo (21%) e Hebdomadis (18%). Dentre as amostras positivas, 14/48 apresentaram títulos de anticorpos aglutinantes superiores a 1:100 e notou-se que 70,83% dos animais soropositivos reagiram a mais de um sorovar de Leptospira interrogans. Somente uma propriedade não possuía vacas sororreagentes para leptospirose e das nove propriedades estudadas, seis (66,66%) apresentaram animais soropositivos ao sorovar Hebdomadis. Ao exame histopatológico, as alterações microscópicas mais encontradas em animais positivos foram hialinização (81,25%), congestão (81,25%) e degeneração hidrópica (70,83%). Porém, essas alterações histopatológicas também foram encontradas em rins de animais negativos à sorologia, como hialinização (80,77%), congestão (48,07%) e degeneração hidrópica (55,77%) e estes achados não apresentaram correlação com positividade. O exame histopatológico dos rins não é indicado para substituir o diagnóstico sorológico da leptospirose, podendo ser utilizado somente como um exame complementar. Apesar da frequência de animais sororreagentes ser baixa na região do Triângulo Mineiro, a doença está presente em um grande número depropriedades. Chama a atenção a alta frequência do sorovar Hebdomadis, podendo ser considerado um sorovar emergente na região. A avaliação da frequência deste sorovar em outras regiões se torna importante, pois uma vez verificada, deve resultar na recomendação da inclusão deste sorovar no controle da leptospirose.

Spatial and temporal dynamics of irrigation water quality under drought conditions in a large reservoir in Southern Portugal.

Water is a scarce resource in the Mediterranean region where adverse climatic conditions promoting water shortages tend to increase with climate change. Under water scarcity conditions and high atmosphere evaporative demand, the risks of decreased water quality, and land salinization are major threats to the sustainability of irrigated agriculture in this region. The assessment of the quality of irrigation water is increasingly important to ensure the maintenance of long-term salt balance at a crop, farm, and regional scale. This study is focused on the spatial and temporal variability of water quality for irrigation in the Alqueva reservoir (Southern Portugal). The assessment was performed every 2 months during a distinctive drought year (2017) and included inorganic ions (Na+, Ca2+, Mg2+, K+, NH4+, Cl-, F-, SO42-, NO3-, and NO2-), pH, and electric conductivity (ECW) of water. Sodium adsorption ratio (SAR) was calculated, and potential soil permeability problems were estimated. The assessment showed significantly higher values of the physicochemical parameters in the most upstream sites, located near tributaries inflows, and an upward trend in ion concentrations throughout the year, with significantly higher concentrations of Na+, Mg2+, Cl-, and SO42, registered through May to November, reflecting the severe drought felt in the summer, autumn, and winter. The evaluation of water quality for irrigation indicated a slight to moderate risk of reduced infiltration rates, which should be considered whenever sprinkler irrigation is used, mainly in fine-textured soils, which are prevalent in the irrigated area. The multivariate statistical approach, using principal component analysis and factor analysis, identified two principal components related to salinity and nutrient concentrations. The cluster analysis revealed three groups of similarity between samples pointing to a more time- than space-controlled pattern. Overall, the temporal dynamics of the water physicochemical parameters could indicate that an abnormal annual distribution of precipitation and temperature may distort seasonal differences. To prevent water and soil degradation, a more frequent assessment of the water quality should be considered, allowing for the selection of appropriate soil and water management measures in irrigated areas.

Use of wastes from the pulp and paper industry for the remediation of soils degraded by mining activities: Chemical, biochemical and ecotoxicological effects.

Fly ash (FA) from biomass combustion and biological sludge (S), both wastes from the pulp and paper industry, were granulated in different proportions (90% FA+10% S, and 70% FA+30% S w/w, dry weight basis, dw) and used to recover the functionality of soils affected by mining activities (Aljustrel, Iberian Pyrite Belt), with and without the application of municipal solid waste compost (MSWC). Application doses of both mixtures were 2.5, 5.0 and 10% (w/w, dw). These materials corrected soil acidity to circumneutral values and increased extractable P and K concentrations. A significant increase in soil organic matter (from 0.6 to 0.8-1.5% w/w, dw) and N content (from 0.04 to 0.09-0.12% w/w, dw) was also observed, but only when MSWC was applied. The soil was already heavily contaminated with Cu, Pb and Zn and the application of amendments did not increase their pseudo-total concentrations. The CaCl2 extractable fractions of both Cu and Zn decreased to very low values. The improvement in soil quality, compared to fertilizer only treatment, was further evidenced by the increase in some soil enzymatic activities (dehydrogenase, ß-glucosidase and cellulase), with a better response for the granules with the higher proportion of biological sludge, as well as by the decrease in the soil-water extract toxicity towards different organisms (Daphnia magna, Thamnocephalus platyurus, and Pseudokirchneriella subcapitata). Agrostis tenuis germinated and grew during the first month only in the amended pots, but, after that, a considerable phytotoxic effect was evident. This was mainly attributed to salt stress or to some specific ionic toxicity. In conclusion, to establish a long-term plant cover in mining soils amended with biomass ash-based materials, the selection of plants with higher resistance to salinity and/or the stabilization of the amendments, to reduce their soluble salt content, is recommended.

Exploring the Use of Species Sensitivity Distributions to Define Protective Limits for the Use of Organic Wastes as Soil Amendments.

The use of organic wastes as soil amendments can be an important measure to improve soil quality and reduce waste accumulation and landfilling. However, the potential contaminant loads of such wastes, can be a source of environmental concern. Consequently, legislation has been developed to regulate the use of these wastes in agricultural soils. However, the regulations only consider chemical parameters, which are insufficient to establish the level of environmental risk. A possible solution is the use of species sensitivity distributions (SSDs), employing ecotoxicological data from test batteries that could be incorporated into legislation. In the present study, 2 different hazardous concentrations affecting 5 and 50% of the soil community (HC5 and HC50, respectively) were determined using ecotoxicological data (effect concentrations, 10 and 50% [EC10 and EC50, respectively]) for 5 different wastes. The results demonstrate that, as expected, current legislative thresholds do not translate to environmental risk/protection and that SSDs may be an important tool allowing the simple inclusion and interpretation of ecotoxicological data from test batteries in legislation. On the other hand, SSDs must be used with caution because there are still doubts about their actual value in risk prediction and about which estimates provide adequate protection. For instance, the use of HC50EC10 values is not recommended; these values overlap with the more conservative HC5EC50 data, highlighting the fact that the use of lower effect concentrations may not always provide the most protective approach. Also, hazardous concentrations need to be calibrated at the field or semifield level, to verify environmental protection in different soils/environments and the adequacy of standard test organisms. Environ Toxicol Chem 2019;38:1569-1576. © 2019 SETAC.

Short communication: Proinflammatory gene expression relative to the collection technique of endometrial samples from cows with and without subclinical endometritis.

Uterine inflammation negatively affects reproductive performance and is an important cause of infertility and subfertility in dairy cows. Several studies have investigated the use of gene expression in endometrial samples collected by biopsy or cytology to evaluate the inflammatory response of the cow uterus. This study aimed to compare the expression of the CCL5, CXCL8, IL6, and IL1B genes in the bovine endometrium according to the site of sample collection [caruncular (C) or intercaruncular (IC)], the collection method (biopsy or cytology), and the category of inflammation based on endometrial cytology (zero, medium, or high) in subclinical endometritis. The reproductive tracts of dairy cows were collected from a slaughterhouse, and punch-biopsy samples of endometrial tissues were obtained from both regions (C and IC). Endometrial cells from these regions were collected with the cytobrush technique and then used for the analysis of mRNA expression by quantitative PCR. After counting polymorphonuclear cells (PMN) by endometrial cytology, 20 uteri with an ovary at stage I (d 1-4 of estrous cycle) were categorized into 3 groups. Uteri with 0% PMN (n = 10) were assigned to group zero, uteri with 5 to 15% PMN (n = 5) to group medium (12.2 ± 1.6% PMN), and uteri with >15% PMN (n = 5) to group high (53.8 ± 32.9% PMN). All data were analyzed with 2-way ANOVA with Bonferroni multiple comparison post test. The results from gene transcripts demonstrated that the region (C or IC) of the endometrial biopsy had no influence on any of the degrees of inflammatory reaction observed. However, gene expression was more elevated in the endometrium of cows with greater inflammation compared with those without inflammation (CCL5, CXCL8, IL6, IL1B) and those with medium inflammation (CCL5, IL6). Expression of the genes evaluated did not differ between the endometrium without inflammation and with medium inflammation. However, in the high inflammation group, all genes were comparatively more expressed in samples collected by cytology relative to those derived from biopsies for both anatomical regions. In conclusion, gene expression did not differ between the C and IC tissue. Samples collected from animals with greater inflammation had greater gene expression than those with zero or medium inflammation. In addition, cytology samples had greater gene expression than biopsy samples in the high inflammation group.

Assessment of the environmental impact of an abandoned mine using an integrative approach: A case-study of the "Las Musas" mine (Extremadura, Spain).

The mine abandonment is generally associated with the release of potentially toxic metals into the environment, which may depend on metals speciation, soil properties and climate conditions. The goal of the present work was to assess the environmental impact of the abandoned Pb-Zn mine "Las Musas" (Spain) using an integrative approach. The impact on soils and surface waters was performed using: chemical parameters, quantification of potentially toxic metals (Cd, Cu, Pb and Zn), and ecotoxicological responses using lethal and sub-lethal bioassays with organisms' representative of different trophic level ((soil: Eisenia fetida (mortality and reproduction test); Latuca sativa and Lollium perenne (seedling emergence); and water: Vibrio fischeri (luminescence inhibition), Daphnia magna (immobility and reproduction test), Thamnocephalus platyurus (mortality), Pseudokirchneriella subcapitata (growth inhibition)). The results showed soils with neutral to slight alkaline pH (7.64-8.18), low electric conductivity (125-953 µS/cm) and low organic matter levels (0.20-1.85%). For most of the soil samples, Pb was the only metal which surpassed the limit proposed by the Canadian soil quality guidelines, with values ranging from 42.2 to 181.4 mg/kg. The ecotoxicological results showed that the soils with the highest levels of Pb induced a decrease on E. fetida reproduction and on L. sativa germination, indicating negative impacts on the habitat function. The analysis of the surface waters showed levels of Zn surpassing the legal limit adopted from the Water Framework Directive (37.0 to 69.0 µg/L). The ecotoxicological results highlight the importance of bioassays that evaluate the behavior of species, when assessing the risk of mining areas with non-acid soils and waters with high nutrients/organic matter concentrations and low concentrations of potentially toxic metals. The results indicated a moderate environmental risk from potentially toxic metals, at the areas analyzed around the Azuaga mine.

Three-dimensional levitation culture improves in-vitro growth of secondary follicles in bovine model.

RESEARCH QUESTION: Does a three-dimensional culture system based on magnetic levitation with nanoparticles assembly maintain the follicular structure and viability with adequate growth rates leading to oocyte maturation after long-term culture? DESIGN: Randomized-controlled trial of treatments in a bovine model. Secondary follicles (n = 213) isolated from bovine ovaries were cultured in a two-dimensional system (two-dimensional control) or three-dimensional levitation system with different concentrations (three-dimensional 50 µl/ml, 100 µl/ml and 200 µl/ml) of magnetic nanoparticles. Follicular growth (diameter, daily growth and growth patterns), morphology (normal, degenerated and extruded follicles), antrum formation, oocyte viability and chromatin configuration were assessed. RESULTS: Secondary follicles of three-dimensional 200-µl/ml treatment showed higher viability, antrum formation and lower degeneration rates than two-dimensional control. Also, follicles cultured in the three-dimensional 200-µl/ml treatment presented a most homogenous daily growth rate as shown by the lowest variance and standard deviation. Compared with the two-dimensional control, the proportion of non-growing and slow-growing follicles were 3.8-fold lower and 1.6-fold higher, respectively, in the three-dimensional 200-µl/ml treatment. After in-vitro maturation, the three-dimensional 200-µl/ml had a greater proportion of viable oocytes (1.7-fold) and meiotic resumption rates (2.4-fold) than the two-dimensional control treatment. CONCLUSION: The three-dimensional levitation culture system improves the viability of in-vitro development of bovine secondary follicles, antrum formation and lower extrusion and degeneration rates and adequate growth rate leading to relevant oocyte viability and meiotic resumption after in-vitro maturation. This approach does not require a specific medium, and has the potential as an alternative method to in-vitro follicle culture in several species, including humans.

Prevalence of inflamatory processes in the reproductive tracts of crossbred dairy cows / Prevalência de processos inflamatórios no sistema reprodutivo de vacas leiteiras mestiças

This study evaluated the prevalence of cervicitis and endometritis and their interrelations in crossbred dairy cows that were more than 60 days in milk (DIM), and the efficiency of the cytology technique (cytobrush) and histology as diagnostic methods for these diseases. The reproductive tracts (n=149) derived from cows were collected from a slaughterhouse and grossly evaluated to determine uterine involution and the phase of the estrous cycle. Subsequently, cervical and uterine cytological evaluations were done to characterize the inflammatory response as cervicitis (with a neutrophilic count greater than 5%) or endometritis (with more than 6% neutrophils). Additionally, randomly obtained samples from the cervix and uterus were collected for histological evaluation. Cytological evaluation revealed that the frequency of cervicitis was 6% (9/149), and endometritis was diagnosed in 8.1% (12/149) of the samples; both inflammatory reactions were diagnosed 2.1% (3/149) in three of these. Histological evaluation revealed that of the cows diagnosed with cervicitis by cytology, 66.6% (6/9) had no inflammatory reaction in the cervical mucosa, and 33.3% (3/9) had mild inflammation in this region. Histological evaluation of the uterine horn revealed that of the 12 cows diagnosed with endometritis by cytology, 50% (6/12) of these had a histological diagnosis of mild and moderate inflammatory responses, 8.3% (1/12) of these demonstrated a severe inflammatory response, while no inflammatory reaction was identified in 41.7% (5/12) of the endometrial biopsies evaluated. In conclusion, the endometrial and cervical inflammatory response diagnosed by the cytobrush technique persisted in 12.1% of the cows. Comparatively, the frequency of the inflammatory responses at the cervix and uterus was reduced by histological analysis relative to cytological evaluation with the cytobrush technique.

O objetivo deste estudo foi determinar a prevalência de cervicite e endometrite citológicas e suas inter-relações em vacas leiteiras mestiças com mais de 60 dias pós-parto (DPP), bem como avaliar as técnicas de citologia (cytobrush) e histologia como métodos diagnósticos para estas doenças. Foram utilizados 149 tratos reprodutivos de vacas com mais de 60 DPP de abatedouro e avaliados macroscopicamente para determinar a involução uterina e a fase do ciclo estral. Posteriormente, realizou-se citologia cervical e uterina para o diagnóstico de inflamação utilizando como ponto de corte, contagem de polimorfonucleares (PMN) superior a 5% e 6%, respectivamente, para cervicite e endometrite citológica. Também foram coletadas amostras de cérvix e útero para histologia. Do total de vacas, identificaram-se 6% (9/149) com cervicite e 8,1% (12/149) com endometrite citológica, sendo três destes animais (2,1%) com ambas as inflamações. Histologicamente, dos animais com cervicite citológica, 66,6% (6/9) não apresentaram infiltrado inflamatório na mucosa cervical e 33,3% (3/9) apresentaram inflamação leve. A histologia uterina mostrou que de 12 vacas com endometrite citológica, 50% (6/12) apresentaram infiltrados inflamatórios leves a moderados, 8,3% (1/12) grave e 41,7% (5/12) não tinham Inflamação endometrial. Concluiu-se que a inflamação endometrial e cervical, por meio da técnica de citologia, persistiu em 12,1% das vacas com mais de 60 DPP. Quando se utiliza a histologia como método de diagnóstico, a detecção de inflamação cervical e uterina é inferior ao detectado pela técnica de citologia por meio do esfregaço endometrial (cytobrush).

Organic wastes as soil amendments - Effects assessment towards soil invertebrates.

Using organic wastes, as soil amendments, is an important alternative to landfilling with benefits to soil structure, water retention, soil nutrient and organic matter concentrations. However, this practice should be monitored for its environmental risk due to the frequent presence, of noxious substances to soil organisms. To evaluate the potential of eight organic wastes with different origins, as soil amendments, reproduction tests with four soil invertebrate species (Folsomia candida, Enchytraeus crypticus, Hypoaspis aculeifer, Eisenia fetida) were performed using gradients of soil-waste mixtures. Results obtained demonstrated that contaminant concentrations required by current legislation might not be a protective measure for the soil ecosystem, as they do not properly translate the potential toxicity of wastes to soil invertebrates. Some wastes with contaminant loadings below thresholds showed higher toxicity than wastes with contaminants concentrations above legal limits. Also, test organism reproduction was differently sensitive to the selected wastes, which highlights the need to account for different organism sensitivities and routes of exposure when evaluating the toxicity of such complex mixtures. Finally this study shows that when combining chemical and ecotoxicological data, it is possible to postulate on potential sources of toxicity, contributing to better waste management practices and safer soil organic amendment products.

Engaging in a Keyhole Concept for the Management of Ruptured and Unruptured Aneurysms.

OBJECTIVE: Many new endovascular devices have been used under the guidance of the International Subarachnoid Aneurysm Trial. Clipping still offers higher occlusion rates, and its technique continues to evolve, resulting in smaller exposures and reduced manipulation to brain tissue. We sought to evaluate the routine use of the minisphenoidal approach to manage intracranial aneurysms in a high-volume institution. METHODS: We retrospectively reviewed our database of patients with aneurysm from October 2013 to May 2016. Data were originally collected prospectively. The minisphenoidal approach has been progressively replacing the pterional approach for managing aneurysms in our department. Occlusion rates for ruptured and unruptured aneurysms were analyzed using late follow-up angiograms. Functional outcome assessment and the impact on quality of life were also measured. RESULTS: We performed 124 minisphenoidal craniotomies in 117 patients to clip 147 aneurysms. Patient mean age was 53.9 years. Seventy patients (59.8%) presented with subarachnoid hemorrhage. Middle cerebral artery aneurysms represented 48% of the total number of aneurysms; posterior communicating artery aneurysms represented 24%. The minisphenoidal craniotomy was helpful in managing superior cerebellar artery aneurysms and 1 ruptured orbitofrontal artery aneurysm. We achieved an occlusion rate of 97.8%, with a mean follow-up of 13.2 months. Favorable outcomes were achieved for 79% of patients with subarachnoid hemorrhage and for 98% of unruptured patients. CONCLUSIONS: Evolution of endovascular techniques has paved the way for minimizing surgical exposures. Routine use of the minisphenoidal approach for managing ruptured, unruptured, and previously coiled aneurysms is safe and provides adequate exposure with robust occlusion rates.