Immunological changes in canine peripheral blood leukocytes triggered by immunization with first or second generation vaccines against canine visceral leishmaniasis

In this study, we summarized the major phenotypic/functional aspects of circulating leukocytes following canine immunization with Leishvaccine and Leishmune?. Our findings showed that Leishvaccine triggered early changes in the innate immunity (neutrophils and eosinophils) with late alterations on monocytes. Conversely, Leishmune(?) induced early phenotypic changes in both, neutrophils and monocytes. Moreover, Leishvaccine triggered mixed activation-related phenotypic changes on T-cells (CD4+ and CD8+ and B-lymphocytes, whereas Leishmune(?) promoted a selective response, mainly associated with CD8+ T-cell activation. Mixed cytokine profile (IFN-γ/IL-4) was observed in Leishvaccine immunized dogs whereas a selective pro-inflammatory pattern (IFN-γ/NO) was induced by Leishmune? vaccination. The distinct immunological profile triggered by Leishvaccine and Leishmune? may be a direct consequence of the distinct biochemical composition of these immunobiological, i.e. complex versus purified Leishmania antigen along with Bacillus Calmette-Guérin (BCG) versus saponin adjuvant. Both immunobiologicals are able to activate phagocytes and CD8+ T-cells and therefore could be considered as a putative vaccines against canine visceral leishmaniasis (CVL).     

Predictive equations of maintenance energy requirement for healthy and chronically ill adult dogs

Maintenance energy requirement (MER) is the energy amount necessary for dogs to maintain their weight and body condition. Some factors can influence the MER, such as gender, age, neutering status and also diseases. The present retrospective study aimed to evaluate MER of adult dogs with several diseases and compare with the MER of healthy adult dogs, observing the influence of parameters such as body condition score (BCS), neutering status, gender, age, diagnosis and type of food on MER of these dogs. A total of 165 adult dogs with weight changes of ≤5% were included and divided in groups according to diagnosis. Mean MER for healthy dogs was 86.09 kcal/BW^0.75, which differed from NRC and FEDIAF recommendations for inactive adult dogs (p = .047). Lowest MERs were of the endocrinopathies (78.52 ± 19.32 kcal/BW^0.75), orthopaedic diseases (59.71 ± 19.30 kcal/BW^0.75) and neurologic diseases (78.83 ± 32.66 kcal/BW^0.75) groups. Gastrointestinal diseases (99.59 ± 20.36 kcal/BW^0.75), orthopaedic diseases (59.71 ± 19.30 kcal/BW^0.75) and neoplasia (95.61 ± 21.02 kcal/BW^0.75) groups were the only groups that differed from the mean MER of healthy adult dogs. Regarding BCS, for each increasing point in a 9-point scale, there was a decrease of 9.8 kcal/BW^0.75 on MER, independent of diagnosis (p < .0001; r² = .55). There was no difference regarding breed size gender and age, but neutered dogs presented lower MER (p = .031). Based on data obtained from the present study, it can be concluded that it is necessary to consider BCS, age, neutering status and diagnosis when calculating MER, both in healthy dogs and chronically ill dogs.     

Species diversity and seasonality of free-living ticks (Acari: Ixodidae) in the natural habitat of wild Marsh deer (Blastocerus dichotomus) in Southeastern Brazil

This study evaluated the presence and seasonal activity of free-living ticks in remaining marsh areas by the Paraná river, in Brazil. Eight field trips (once per season) for collection of ticks were performed during 2 years. Using CO2 traps, dragging, and visual inspection of vegetation, five free-living tick species were collected, in the following order of abundance: Amblyomma cajennense, Amblyomma dubitatum, Amblyomma triste, Amblyomma coelebs, and Amblyomma nodosum. The seasonal pattern of A. cajennense was characterized by the highest peaks for adult ticks in the summer/spring months, for nymphs in the winter and for larvae in the autumn and winter. A. dubitatum and A. triste presented similar seasonal patterns characterized by peaks of adult ticks in the autumn. Nymphs of A. dubitatum peaked in the winter of the first year and in the winter/spring of the second year. A. triste was the only species to be collected in significantly higher numbers in the marsh than in surrounding drier areas such as forest patches. Among domestic animals living close the marsh areas, horses were infested by Anocentor nitens, A. cajennense, and Boophilus microplus, bovines were infested solely by B. microplus, and dogs were infested by Rhipicephalus sanguineus. Adults of A. triste showed to be well adapted to the marsh environment. This result, at least partially, explains local previous observations on the association of A. triste with marsh deer, as this vertebrate host inhabits mainly the marsh area.     

Detection and quantification of provitamin D2 and vitamin D2 in hop (Humulus lupulus L.) by liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry

In this work, ergosterol and ergocalciferol were identified for the first time in hop. In addition, in this article, a simple and reliable analytical methodology for analysis of these compounds in different commercial forms of hop is presented. The performance of the method was assessed by the evaluation of parameters such as absolute recovery (higher than 70%), repeatability (lower than 3 %), linearity ( r(2) > 0.9988) and limits of detection (ranging from 0.034 for ergocalciferol to 0.058 mg/L for ergosterol) and quantification (ranging from 0.113 for ergocalciferol to 0.195 mg/L for ergosterol). On the basis of standard additions applied with the optimized procedure and high-performance liquid chromatography with diode array detection, it appears that the Nugget hop plant (crop 2006) contains 1.84 +/- 0.09 microg/g of ergosterol and 1.95 +/- 0.05 microg/g of ergocalciferol. The identity of the compounds was confirmed by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry in the positive ion mode. The presence of ergosterol here reported should have great potential for the assessment of hop as related to the fungal contamination proportion and hence the quality of this raw material.     

Biodegradation of biodiesel and toluene under nitrate-reducing conditions and the impact on bacterial community structure

Purpose

Biodiesel is a renewable fuel that can be mixed with toluene and be accidentally released into anoxic ecosystems and impact soil microbial communities. Therefore, the aim of the present work was to examine, under nitrate-reduction conditions, the biodegradation of toluene in the presence of two different types of biodiesel (sunflower and rapeseed), and their impact on the bacterial community structure.

Materials and methods

Sediment samples were spiked individually with toluene, biodiesel, and their blends in laboratory-designed microcosms. Sunflower oil biodiesel was produced in the laboratory, while rapeseed oil biodiesel was a commercial product. Degradation of biodiesels and blends was monitored by directly measuring the substrate or indirectly by determining nitrate removal during the course of the experiment. Denitrification rates were estimated with the acetylene inhibition technique. Denaturing gradient gel electrophoresis was used to assess impacts on the bacterial community structure exposed to biodiesels, blends, and toluene.

Results and discussion

The results of this study showed that toluene and biodiesel were completely degraded within 10 days. Biodiesel significantly affected the bacterial community structure at a similar magnitude, independently of its origin. Additionally, toluene impacted the bacterial community and denitrification process to a lower extent than biodiesel and a clear decrease in the relative bacterial richness and diversity was shown in samples with biodiesel and blends. To the best of our knowledge, this is one of the first reports describing degradation of biodiesel alone and blends under nitrate-reducing conditions, and also the effects of these compounds on the denitrification process. In addition, due to the recently discovered “oxygenic denitrification” process, the acetylene inhibition technique and nitrous oxide quantification may not be the most adequate tool to estimate denitrification rates. Further detailed studies are advised to understand whether the identified bacterial community shift impacts ecosystem functions.

Conclusions

Our results help to understand the biodegradation of toluene, biodiesel, and their blends in sediments under nitrate-reducing conditions and might be important in implementing bioremediation strategies in anoxic environments.

     

Potential environmental impact of technosols composed of gossan and sulfide-rich wastes from São Domingos mine: assay of simulated leaching

Purpose

The aim of this study was to evaluate (i) the ability of two Technosols, prepared with gossan or sulfide-rich wastes plus mixtures of organic/inorganic amendments, to improve the characteristics of the wastes by the analysis of the variation of elemental concentrations in their simulated leachates, and (ii) the potential environmental risk of these Technosols evaluated through the concentrations of the elements leached from the tailings containing the two wastes, considering their mass in the São Domingos mine.

Materials and methods

Composite samples of two São Domingos mining wastes (gossan wastes—GW; sulfide-rich wastes—SW) were collected. Amendment mixtures, containing different organic/inorganic wastes (from green agriculture, distillation of Ceratonia siliqua and Arbutus unedo fruits, and limestone quarry), were applied at 12, 30, and 60 g/kg. Two sets of microcosm assays were performed under controlled conditions in greenhouse and monitored during 7 and 13 months for GW and SW, respectively. Materials from each pot/treatment (<5 cm of depth) were collected after 1, 4, 7, and 13 months of incubation and used to obtain simulated leachates (DIN extraction). The analytical parameters evaluated in the leachates were pH, electrical conductivity, and element concentrations using flame atomic absorption spectrometry for cations and graphite furnace atomic absorption spectrometry, ionic chromatography, and UV-VIS for elements that normally occur as anions in aqueous solution.

Results and discussion

Simulated leachates from SW had lower pH and higher concentrations of potentially hazardous elements than leachates from GW. The concentrations of As in leachates from GW-Technosols were higher than those in leachates from GW-control but <0.6 mg/kg. In GW-Technosols leachates, the pH and the concentrations of some nutrients (e.g., Ca and phosphates) also increased when compared to the control. In the SW-Technosols, the pH of the leachates increased only in the first month as long as limestone was present. In these leachates, a general decrease of the concentrations of some elements was observed (e.g., As, sulfate, Fe, Pb), especially in the first month. A clear influence of the dose and type of amendments was not observed during the experimental time span for both wastes.

Conclusions

The concentrations of elements like Al, Ca, Fe, and Pb in the leachates are controlled by both the pH of the solutions and the concentrations of phosphates and sulfates. The extensive mass of the studied mine wastes contribute to the release to the neighboring environment of considerable amounts of potentially hazardous elements. The rehabilitation of the mine wastes by the conception of Technosols, especially with sulfide-rich wastes, can reduce significantly their environmental impact.

     

Human disturbance affects enzyme activity,microbial biomass and organic carbon in tropical dry sub-humid pasture and forest soils

ABSTRACT

The Brazilian dry forest occupies an area of about 1 million km² approximately 46% of which has been deforested. Many studies have been done on the effects of this on productivity and soil chemical attributes. However, little is known about soil enzymatic activity, which is sensitive to environmental changes. The objective of this study was to evaluate the effects of deforestation using different levels of human disturbance on soil enzyme activities, organic carbon content, microbial biomass, and microbial community. We studied areas covered with forest (TDF), old grass (OG), or new grass (NG). Soils from NG had increased microorganisms, which restored important processes related to carbon, sulfur, and nitrogen cycling, so that they resembled those in the forested area. The results of this study showed that the conversion of forest to pasture with a high level of human disturbance could decrease the activities of β-glucosidase, urease, alkaline phosphatase, and fluorescein diacetate in the soil by up to 87%, 66%, 62%, and 58%, respectively. These findings suggest that human disturbance can cause substantial changes in the enzymatic activity and microbial community in the soil. We suggest that maintaining grass pasture with low human disturbance should have fewer impacts on soil quality.     

Risk assessment of Arbutus unedo L. fruits from plants growing on contaminated soils in the Panasqueira mine area,Portugal

Purpose

In the Panasqueira mine area, Arbutus unedo L. (arbutus tree) grows on soils developed on waste materials and on soils impacted by mining activity. The arbutus berry brandy is considered a product with economic value. The aims of this study were to evaluate the biogeochemical impact of the mining activity on soils and arbutus trees, to assess the possible risks associated with human consumption of the fruits and the derived brandy, and to evaluate the potential of the arbutus tree in phytostabilization.

Materials and methods

Soil samples (10–15 cm deep) developed on waste materials, on schists affected by seepage water or treatment plant effluents and on colluvium-alluvium materials were characterized (fraction <2 mm) for pH, particle size distribution, organic carbon (C_org), cation exchange capacity (CEC) and NPK by classical methodologies. Plant (A. unedo) samples (roots, leaves and twigs, and fruits) were collected at the same sites as the sampled soils, washed with tap and distilled water and dried at 40 °C. The elements’ concentrations in soils (total fraction—four-acid digestion and available fraction—diethylenetriaminepentaacetic acid extraction), plants (ashing followed by acid digestion) and brandy samples produced with fruits collected on contaminated and non-contaminated sites were determined by inductively coupled plasma atomic emission spectroscopy.

Results and discussion

The soils are mainly acid, silty loam, with variable values for C_org, CEC and NPK. They are contaminated with As (158–7,790 mg/kg), Cd (0.6–79 mg/kg), Cu (51–4,080 mg/kg), W (19–1,450 mg/kg) and Zn (142–12,300 mg/kg). The available fraction of the soils is quite variable between <0.04 and 76 % of the total, depending on the element. Trace elements’ concentrations, in leaves and twigs, are within the normal range for plants, except for Cd and Zn that, in some samples, are above the normal values, but without phytotoxic symptoms. Trace elements’ concentrations in fruits are low. The calculated hazard quotient for all trace elements in arbutus berry was <0.1. In the brandy, elemental concentrations are within the legal standards, except for Pb, whose higher concentrations may result from distillery equipment.

Conclusions

According to the EC 466/2001 legislation and with a hazard quotient of <1, the arbutus berry consumption does not constitute health risks for humans. The fruits can be used to produce local brandy. The concentration of copper in brandy is within the range established by the Portuguese legislation. Arbutus unedo can be used in the phytostabilization programs in the Panasqueira area, for it is a pioneer species and a non-accumulator of trace elements.     

Inter-population variation on the accumulation and translocation of potentially harmful chemical elements in Cistus ladanifer L. from Brancanes,Caveira, Chança,Lousal, Neves Corvo and São Domingos mines in the Portuguese Iberian Pyrite Belt

Purpose

This study aimed to compare the variation on the accumulation and translocation of potentially harmful chemical elements and nutrients (As, Ca, Cu, Fe, K, Mg, Mn, Ni, Pb and Zn) in Cistus ladanifer L. belonging to populations growing in different mine areas from the Portuguese Iberian Pyrite Belt (Brancanes, Caveira, Chança, Lousal, Neves Corvo, São Domingos). These mines are abandoned (except Neves Corvo that is still operating) and have different contamination levels.

Materials and methods

Composite samples of soils (n?=?31), developed on different mine wastes and/or host rock, and C. ladanifer plants (roots and shoots) were collected in the mine areas. Soils were characterized for pH, NPK and organic C, by classical methodologies. Soils (total fraction—four acid digestion, and available fraction—extracted with aqueous solution of diluted organic acids, simulating rizosphere conditions) and plants (ashing followed by acid digestion) elemental concentrations were determined by ICP. Soil–plant transfer and translocation coefficients were calculated. Principal components analysis in both ways, the classical method and a second approach with adaptations used mostly in multivariate statistical processes control data, were done in order to compare the plants populations.

Results and discussion

Soils had large heterogeneity in their characteristics. Caveira, Lousal, Neves Corvo and São Domingos soils showed the highest total concentrations of As, Cu, Pb and Zn. Independently of the mine, available fractions of elements were low. Intra- and inter-population variations in accumulation and translocation of elements were evaluated. Plants were not accumulators of the majority of the analysed elements. Nutrients were mainly translocated from roots to shoots, while trace elements were stored in roots (except in Neves Corvo for As and Pb, and São Domingos for As). Elements concentrations in plant populations from Lousal, Chança and São Domingos did not present much variation. Brancanes soils and plants presented strong differences compared to other areas.

Conclusions

Cistus ladanifer plants are able to survive in mining areas with polymetallic contamination at different elements concentrations in total and available fraction. This species presented variations inter- and intra-populations in accumulation and translocation of chemical elements; however, all studied populations, except Brancanes, can belong to the same population cluster.