Application of a plant bioassay for the evaluation of ecotoxicological risks of heavy metals in sediments affected by mining activities

Purpose

The objective of this work was to evaluate the effectiveness of a plant bioassay (Phytotoxkit®) for screening ecotoxicological risks in sediments affected by mining activities.

Materials and methods

A total of 42 sediment samples affected by mining activities were studied, including 39 sediment samples from the Sierra Minera, Spain, an area affected by old extraction procedures, and three sediments from an area affected by opencast mining. These three samples were then mixed with limestone filler at 10, 20 and 30 %, providing nine stabilised samples. The total and soluble metal(loid) content (As, Cd, Cu, Fe, Pb and Zn) was determined in all samples, and the Phytotoxkit® bioassay was applied to determine the ecotoxicological effect of this procedure.

Results and discussion

The stabilised material had a neutral pH and low soluble metal(loid) concentration, similar to that of samples in which a natural attenuation process had taken place because of mixing with surrounding carbonate-rich materials. An ecotoxicological survey identified the low toxicity levels of the stabilised samples.

Conclusions

The applied bioassay is a good tool for screening metal(loid) contamination in areas affected by mining activities, since it provides information on both natural and simulated attenuation processes. The mixing of sediments with limestone filler could be applied to the remediation of zones affected by mining activities, because the toxicological effect on the tested organisms in the stabilised sediments was reduced significantly and the metal(loid) content was diminished.     

Quantification of mucosa-adhered microbiota of lambs and calves by the use of culture methods and fluorescent in situ hybridization coupled with flow cytometry techniques

The intestinal mucosa-associated microbiota could play important biological roles due to its close proximity with the animal host, but knowledge on its composition is still limited. The aim of this study was to characterize the microbial communities tightly associated with different parts (rumen, duodenum and colon) of the gastrointestinal tract (GIT) of healthy lambs and calves by using both cultural, and fluorescent in situ hybridization-flow cytometry (FCM-FISH) techniques. Lactic acid bacteria genera were one of the predominant bacteria detected in lambs and calves by both methodologies, possibly constituting an index of their healthy status. The levels of Lactobacillus were significantly higher (p<0.05) in the rumen and duodenum of lambs, and in the rumen of calves. The levels of Bifidobacterium were significantly higher (p<0.05) in the colon of both animal species and the rumen of lambs. Significant differences (p<0.05) were found in counts of other microbial groups (yeast, Enterococcus, Propionibacterium, Bacteroides, Clostridium and Enterobacteriaceae) at diverse GI sections depending on the animal species. In general, microbial counts follow the same trends regardless the applied technique. The most remarkable differences were found in detection levels of Bacteroides and Clostridium, which tended to be significantly higher (p<0.05) when analysed by FCM-FISH. This technique also allowed the detection of quantitatively important bacteria (sulphate-reducing bacteria, Atopobium and Coriobacterium), which are difficult to cultivate in selective medium. Therefore, FCM-FISH has been proven to be a sensitive high throughput approach that provides additional information to that obtained by traditional culture techniques about the complexity of the GI ecosystem of these animal species.     

Phenytoin as a novel anti-vitiligo weapon

Vitiligo is a psychologically devastating clinical conundrum which affects approximately 1% of the general population. The exact cause of the illness is an enigma, but several hypotheses about its pathogenesis are advanced.The autoimmune hypothesis proposes an autoimmune attack against melanocytes. Although anti-melanocyte autoantibodies have been demonstrated in vitiligo, recent research casts doubt on their pathogenic role and instead supports the involvement of cell-mediated autoimmune response in the pathobiology of this disorder, which is characterized by increase of suppressor T-cells and decrease of the helper/suppressor ratio in association with the presence of type-1 cytokine secreting cytotoxic T cells in the vicinity of disappearing melanocytes.The neural hypothesis proposes that increased release of norepinephrine, a melanocytotoxin, from the autonomic nerve endings in the microenvironment of melanocytes injures these cells. Moreover, norepinephrine induces the catecholamine degrading enzyme monoamine oxidase (MAO), which favors the formation of toxic levels of hydrogen peroxide in the vicinity of melanocytes.Another theory suggests that abnormal permeability of melanosome membrane, which normally prevents the diffusion of toxic melanin precursors into the cytoplasm, may cause melanocyte damage.Phenytoin, the widely-used anticonvulsant, has been employed both topically and systemically in the treatment of some dermatological disorders. The drug has been shown to significantly suppress mitogen-induced activation of lymphocytes and cytotoxic T lymphocyte activity and to polarize the immune response toward the type-2 pathway. It also significantly decreases suppressor T cells and increases the helper/suppressor ratio.At high concentrations, the drug inhibits the release of norepinephrine and the activity of MAO. Moreover, phenytoin is suggested to interact with membrane lipids, which may promote stabilization of the membranes.The hydantoin moiety of phenytoin exerts a direct stimulatory action on melanocytes; facial hyperpigmentation is a recognized side effect of orally administered phenytoin.Altogether, the above evidence suggests that phenytoin could be therapeutically effective against vitiligo. As phenytoin stimulates collagen production and inhibits its breakdown, its concomitant use with topical steroids could prevent steroid-induced skin atrophy while potentiating the anti-vitiligo effect of these agents.     

Identification,potential inoculum sources and pathogenicity of botryosphaeriaceous species associated with grapevine dieback disease in New Zealand

This study investigated the prevalence and identity of botryosphaeriaceous dieback pathogens in necrotic grapevines tissues in New Zealand vineyards, and other woody hosts growing nearby. The presumptive identities of the isolates by conidial and cultural morphology were confirmed with ITS sequence data as Neofusicoccum australe, N. luteum, N. parvum and Diplodia seriata. They were isolated predominantly from necrotic stems of grapevine and other hosts, but also from leaves, flowers and wood debris of grapevines. Inoculation with conidia and mycelium of multiple isolates of each species onto excised and attached green shoots and trunks of five grapevine varieties, Cabernet sauvignon, Chardonnay, Pinot noir, Riesling, and Sauvignon blanc, showed that all varieties became infected to a similar extent. All species except D. seriata were pathogenic, irrespective of the host source, with N. luteum being the most and D. mutila the least pathogenic (P < 0.05). On trunks, N. parvum caused cankers and the other pathogenic species caused die-back when the inoculated vines became winter-dormant. Conidia were produced from green shoot lesions and die-back wood, which indicates potential inoculum sources for vineyard infection.     

Artificial long-day photoperiod in the subtropics increases milk production in goats giving birth in late autumn

Two experiments were conducted to determine whether exposure to a photoperiod of artificial long days in autumn increased milk yield in subtropical goats milked once (Exp. I) or twice daily (Exp. II). In Exp. I, starting at d 10 of lactation, 1 group of does was kept under naturally decreasing photoperiod (DD1X; n = 8), whereas the other group was submitted to an artificial photoperiod of long days (LD1X; n = 8; 16 h light:8 h darkness). The kids were weaned 28 d after parturition, and dams were manually milked once daily. Milk yield and milk components (fat, protein, and lactose) were assessed up to 140 d of lactation. From d 0 to 28 of lactation (suckling phase), mean daily milk yield did not differ between DD1X and LD1X goats (2.3 ± 0.2 kg vs. 2.4 ± 0.2 kg; P = 0.717). However, between d 29 and 84 (early milking phase), mean daily milk yield was greater in LD1X does than in DD1X does (2.6 ± 0.1 kg vs. 2.1 ± 0.1 kg; P = 0.001). Finally, between d 85 and 140 (late milking phase), mean daily milk yield was greater in LD1X goats than in DD1X goats (P ≤ 0.05) only during the first 2 wk. In Exp. II, one group of goats was exposed to a photoperiod of naturally decreasing days (DD2X; n = 8) and another group was submitted to an artificial photoperiod of long days (LD2X; n = 7). In both groups, kids were weaned on d 28 of lactation and the dams were manually milked twice daily. During the nursing phase, mean daily milk yield did not differ between the DD2X and LD2X groups (2.5 ± 0.3 kg vs. 2.6 ± 0.2 kg; P = 0.767). In the early milking phase, mean daily milk yield was greater in LD2X than in DD2X goats (3.3 ± 0.2 kg vs. 2.8 ± 0.2 kg; P = 0.022), whereas during the late milking phase, milk yield did not differ between the 2 groups (P = 0.946). In both experiments, milk composition was not significantly influenced by exposure to long-day photoperiod. We conclude that, in subtropical female goats that start lactation in late autumn, exposure to an artificial long-day photoperiod stimulates milk yield, even if goats are milked once daily. In addition, combining exposure to long days with twice-daily milking will increase further milk yield in such goats without affecting milk components.     

In Vivo and In Vitro Sperm Interaction with Oviductal Epithelial Cells of Llama

Sperm reservoirs in South American Camelids would be crucial for successful fertilization. Since ovulation occurs approximately 36 h after mating, the maintenance of the sperm viability in the oviduct waiting for the ovum is a critical reproductive event. Our study aimed at determining whether the isthmus or the utero tubal junction (UTJ) could function as a sperm reservoir in llama by means of in vivo and in vitro experiments. For the in vivo experiments, the oviducts of adult females with a dominant follicle bigger than 7 mm were examined for the presence of sperm at 6, 18, 24, 28 and 35 h after mating. The results using scanning and transmission electron microscopy showed ultrastructural differences between isthmus and UTJ with respect to (1) predominance of secretory cells in the UTJ and ciliated cells in the isthmus epithelium and (2) cytoplasmic bulbous projection of the secretory cells in the UTJ. Sperm adhered by a mucus‐like substance were seen only in the UTJ at 6, 18, 24 and 28 h post‐mating. Lack of sperm adhered to oviductal mucosa was observed around ovulation (35 h). In vitro experiments demonstrated higher ability of UTJ epithelial cell explants with respect to isthmus explants to bind sperm in a co‐cultured system. The anatomical features and the presence of a sperm bonding agent in the UTJ together with the in vitro differential binding of sperm to UTJ explants strongly suggest that both may be feasible mechanisms that facilitate sperm storage in this oviductal region in llama.     

Model identification of the bundle thickness dynamics in drum friction spinning

The dynamics of the friction yarn formation process is described in terms of the dynamic behavior of the process state variables. Production efficiency and product quality of friction spinning are influenced by these process state variables. This study was performed with the intention of setting up a dynamic model based on mass balance in order to delineate the dynamic behavior of the cross-sectional area of the in-process bundle on the friction drum surface and to attain the distribution profiles of the cross-sectional area in transient state and in steady state as well. Then, simulation results were compared with those of the experiment, while the input variation was used as a test signal. Theoretical model shows that the bundle thickness on the friction drum has the dynamic characteristics of an integrating system with interval due to the finite length of the input fleece zone, and is in good correspondence with the results of the experiment. A periodic change in the fleece feeding rate led to the fact that there was a specific range of the fluctuation wavelength in the fleece feeding rate, whereas a change in the in-process bundle thickness completely disappeared or became locally maximized in certain wavelength areas. This could lead to a magnification or inaction of the perturbations with the corresponding wavelength in the bundle thickness; a bundle thickness irregularity. Experiments also identified that the theoretical model accurately described the fiber accumulation dynamics in a friction spinning process.