Phytoremediation of metals using lemongrass (Cymbopogon citratus (D.C.) Stapf.) grown under different levels of red mud in soil amended with biowastes

Due to hostile condition of red mud (RM), its utilization for vegetation is restricted. Therefore, RM with biowastes as soil amendment may offer suitable combination to support plant growth with reduced risk of metal toxicity. To evaluate the effects of RM on soil properties, plant growth performance, and metal accumulation in lemongrass, a study was conducted using different RM concentrations (0, 5, 10, and 15% w/w) in soil amended with biowastes [cow dung manure (CD) or sewage-sludge (SS)]. Application of RM in soil with biowastes improved organic matter and nutrient contents and caused reduction in phytoavailable metal contents. Total plant biomass was increased under all treatments, maximally at 5% RM in soil with SS (91.4%) and CD (51.7%) compared to that in control (no RM and biowastes). Lemongrass acted as a potential metal-tolerant plant as its metal tolerance index is >100%. Based on translocation and bioconcentration factors, lemongrass acted as a potential phytostabilizer of Fe, Mn, and Cu in roots and was found efficient in translocation of Al, Zn, Cd, Pb, Cr, As, and Ni from roots to shoot. The study suggests that 5% RM with biowastes preferably SS may be used to enhance phytoremediation potential of lemongrass.     

DNA isolation protocol for red seaweed (rhodophyta)

We report a DNA isolation protocol for red seaweed. The method is a modification of the Dellaporta et al. (1983) protocol for land plants. Our simplified version can be used to process large sample numbers and to minimise polysaccharide co-isolation. The protocol was applied to 12 red seaweed species as well as one green alga and one land plant. The protocol yields about 5 μg of high molecular weight DNA from 10 mg of dried material, with no RNA. No sign of degradation was observed after agarose gel electrophoresis for both freshly extracted DNA and DNA stored for 18 months at 4°C. DNA isolated by our protocol was suitable for genomic library construction (tested for one species), endonuclease restriction, and PCR amplification for all species.     

Modelling sand/mud transport and morphodynamics in the Seine river mouth (France): an attempt using a process-based approach

The mouth of the Seine River estuary (France) has undergone marked morphological evolution over several decades mainly due to engineering works aimed at improving access to Rouen and Le Havre harbours. The intertidal areas are decreasing in size and the lower estuary is accumulating sediment and prograding. In order to understand and better describe the major morphological behaviours of the estuary, a morphodynamic numerical model was developed within the Seine-Aval program. At the end of the 1st part of the research program, a validated fine sediment transport model (3D) was available (Le Hir et al., 2001b). As the present morphological study addresses medium-term issues (a few decades), and because of the need to investigate impacts of local structures or events, we chose to use the so-called “process-based approach” starting from the existing model. First, the existing model was upgraded to account for (suspended) sand transport, and to achieve coupling between morphological changes and sediment transport. Erodability of the sediment accounts for the respective proportions of mud and sand. Simulations starting from an arbitrary surficial sediment cover show that the model is able to reproduce realistic sediment patterns. For example, it is able to change the sediment nature on the intertidal flat near Le Havre from sand to mud. Observed structures of suspended sediment are also reproduced: fine particles mainly follow the turbidity maximum whereas significant concentrations of sand grains in suspension are found where the hydrodynamic stresses are intense. Concerning morphodynamics, simulations with real forcing over one year are discussed. The effect of waves on the bathymetric evolution of the mouth is shown and the sensitivity of morphodynamics to the coupling procedure is tested.     

Parasites reduce territorial behaviour in red grouse (Lagopus lagopus scoticus)

Hypotheses for the cause of population cycles have focused on the role of intrinsic mechanisms such as spacing behaviour and extrinsic mechanisms such as parasitism and predation. This paper examines the interaction between the two dominant hypotheses in the cycles of red grouse, parasitism and spacing behaviour. The influence of the caecal nematode, Trichostrongylus tenuis, on male aggressiveness was investigated using two approaches. First, the territorial behaviour of a group of male grouse experimentally treated with an anthelmintic to reduce parasite intensities was compared with a control group with natural intensities of infection. Second, the response of treated and control males to a novel conspecific territorial intruder were recorded using playback tests. Treated males, with reduced levels of parasitism, won significantly more territorial contests than control males and produced more aggressive behaviour in response to the playback recordings. These results show that parasite removal enhanced aggressive behaviour. Implications of these results on red grouse population dynamics are discussed.     

Bullet-derived lead in tissues of the wild boar (Sus scrofa) and red deer (Cervus elaphus)

Bullet-derived lead in game food products is an important source of human contamination. Careless removal of tissues from around the bullet pathway in the animal body results in elevated lead doses being ingested by humans. To assess bullet-derived lead contamination of soft game tissues, muscle tissue samples were collected from ten wild boars and ten red deer immediately after they had been shot. The samples were collected from around the entry and exit wounds, from around the bullet pathway at different sites along its length, and from a distance of about 5, 15, 25, and 30 cm from the bullet track. The individuals examined differed in the lead contents in their tissues surrounding the entry and exit wounds and at different sites along the bullet pathway. One of the animals showed as much lead as 1,095.9 mg kg⁻¹ wet weight in the tissue surrounding the bullet track near the entry wound, 736.0 mg kg⁻¹ being recorded around the exit wound.     

Palynological evidence for Late Miocene-Pliocene vegetation evolution recorded in the red clay sequence of the central Chinese Loess Plateau and implication for palaeoenvironmental change

In northern China, the Late Miocene-Pliocene red clay in the eastern Loess Plateau fills a gap of climate records between the well-known loess-soil sequences of the last 2.6 Ma and the Miocene loess-soil sequences from the western Loess Plateau. Previous studies indicate that the red clay is also of wind-blown origin, covering the period from ∼ 7-8 to ∼ 2.6 Ma. The red clay therefore provides a good archive to reconstruct paleoecological succession and paleoclimate change. In this study, a palynological investigation was conducted on the late Miocene-Pliocene red clay sequence at Xifeng, central Loess Plateau, which provides new insights into the nature of the evolution of vegetation and climate change from ∼ 6.2 to ∼ 2.4 Ma. Our results show that during this period the central Loess Plateau region was covered mainly by a steppe vegetation, indicating long lasting dry climatic condition. Three vegetational zones were recognized during this period. Zone A (∼ 6.2 to ∼ 5.8 Ma) is characterized by a steppe ecosystem; Zone B (∼ 5.8 to ∼ 4.2 Ma) is characterized by a significant increase of temperate forest plants, indicating a relatively humid regional climate; Zone C (∼ 4.2 to ∼ 2.4 Ma) indicates a typical steppe ecosystem. The vegetation shift at about 4.5-3.7 Ma, when the temperate forest plants decrease, the vegetation gradually changed to typical grassland and even to desert steppe. This is interpreted to represent a drying event. The uplift of the Tibetan Plateau at about 4.5 Ma that resulted in the intensification of the monsoon reversal is thought to have played an important role in this significant ecological change. High-latitude cooling may have partially contributed to the climate shift during ∼ 4.5 to ∼ 3.7 Ma in the Loess Plateau region, and most likely was the driving force for the ecological shift at about 3.7 Ma.     

Use of the FlowCAM for semi-automated recognition and enumeration of red tide cells (Karenia brevis) in natural plankton samples

Early detection is the most effective way to mitigate the effects of harmful algal blooms (HAB). Cell counts based on examination of microplankton samples using settling chambers and visual inspection with an inverted microscope are tedious and time consuming, and counting precision is generally poor at low cell densities. The FlowCAM is a continuous imaging flow cytometer designed to characterize particles in the microplankton size range (20–200 μm diameter). In this study we examined the ability of the FlowCAM to improve routine monitoring protocols for HAB species by automatically recording information on size and fluorescence per cell. This will eliminate the need to examine cells outside the ranges of these measurements for our target species, Karenia brevis. We also tested the ability of image comparison software to match images of cells in mixed assemblages to images of the target species. For simple mixtures of cultured dinoflagellates, the ability of the image matching software to discriminate target cells varied greatly depending on how similar the two species were in size and shape. When target cells were added to natural plankton samples, the image recognition software correctly identified 80–90% of the target cells, but misidentified 20–50% of non-target cells in the size range of the target species. We conclude that the FlowCAM is less tedious and time-consuming than microscopy, allowing for examination of more cells for greater counting precision. The cell recognition software helps reduce the numbers of cells that must be screened, but images must still be examined by a trained operator to identify the HAB species of interest.     

Red mud (RM)-Induced enhancement of iron plaque formation reduces arsenic and metal accumulation in two wetland plant species

Human activities have resulted in arsenic (As) and heavy metals accumulation in paddy soils in China. Phytoremediation has been suggested as an effective and low-cost method to clean up contaminated soils. A combined soil-sand pot experiment was conducted to investigate the influence of red mud (RM) supply on iron plaque formation and As and heavy metal accumulation in two wetland plant species (Cyperus alternifolius Rottb., Echinodorus amazonicus Rataj), using As and heavy metals polluted paddy soil combined with three rates of RM application (0, 2%, 5%). The results showed that RM supply significantly decreased As and heavy metals accumulation in shoots of the two plants due to the decrease of As and heavy metal availability and the enhancement of the formation of iron plaque on the root surface and in the rhizosphere. Both wetland plants supplied with RM tended to have more Fe plaque, higher As and heavy metals on roots and in their rhizospheres, and were more tolerant of As and heavy metal toxicity. The results suggest that RM-induced enhancement of the formation of iron plaque on the root surface and in the rhizosphere of wetland plants may be significant for remediation of soils contaminated with As and heavy metals.     

A suitable extractant for assessing plant-available copper in different soils (peaty,red and alluvial)

Summary Relative efficiency of five chemical extractants for the extraction of available Cu in four different soils and its uptake by rice seedlings was studied in a pot culture experiment. The Cu extracted by 0.1N HCl showed significant relationship with plant uptake whereas the values for other extractants except 0.05M EDTA did not approach the level of significance. Among the soil properties studied, pH and CaCO₃ correlated negatively and organic matter and CEC positively with Cu uptake by rice plants.     

Selenium concentration in liver and kidney of free living animals (roe and red deer) from West Pomerania (Poland)

The aim of this study was to determine concentrations of selenium in the liver and kidneys of roe deer and red deer from West Pomerania, depending on the season. Altogether, samples from 169 animals were collected (96 from roe deer and 73 from red deer) in 2003–2007. The mean concentration of selenium in the liver of red deer and roe deer was 0.37 μg/g and 0.62 μg/g dry weight, respectively. In kidneys, Se concentration was 2.72 μg/g d.w. in red deer and 2.99 μg/g d.w. in roe deer. In roe deer, liver selenium concentration in autumn was significantly higher than in winter (P < 0.05) and spring (P < 0.01) and significantly lower in spring than in summer (P < 0.05); likewise, kidney selenium concentration was higher in autumn than in summer. In deer, no statistically significant season-related differences were observed for liver selenium concentrations. In red deer kidneys, selenium concentration was the lowest in summer, significantly lower than in autumn and winter. Low selenium concentrations in the analyzed tissues show that the animals live in areas deficient in this element.     

High genetic diversity and low population structure in red foxes (Vulpes vulpes) from Croatia

The red fox (Vulpes vulpes) is a highly adaptable omnivorous mammal distributed across all continents on the northern hemisphere. Although the red fox is present throughout Europe, where it plays an important ecological and socio-economic role not only as a game species but also as a rabies reservoir, few studies have examined its population-level mitochondrial DNA variability. In this study, 27 mitochondrial DNA control region haplotypes were identified in 229 red fox samples taken from four regions in Croatia. Haplotype diversity of Croatian red foxes (0.901) was found to be among the highest of all European red fox populations studied to date. Genetic differentiation among regions was quite low, and statistically significant estimates of differentiation were obtained only when comparing the population from the peninsular region of Istria with the three continental populations. It seems that landscape barriers like rivers and small mountains do not restrict gene flow among foxes in the continental part of Croatia, while the combination of a narrow land bridge and altitudes exceeding 1000 m limit fox migration between Istria and the rest of the continent. Better understanding of small-scale population structure will require analysis of highly variable nuclear markers like microsatellites.     

Biodiesel production via transesterification of palm olein using waste mud crab (Scylla serrata) shell as a heterogeneous catalyst

A recent rise in crab aquaculture activities has intensified the generation of waste shells. In the present study, the waste shells were utilized as a source of calcium oxide to transesterify palm olein into methyl esters (biodiesel). Characterization results revealed that the main component of the shell is calcium carbonate which transformed into calcium oxide when activated above 700 °C for 2 h. Parametric studies have been investigated and optimal conditions were found to be methanol/oil mass ratio, 0.5:1; catalyst amount, 5 wt.%; reaction temperature, 65 °C; and a stirring rate of 500 rpm. The waste catalyst performs equally well as laboratory CaO, thus creating another low-cost catalyst source for producing biodiesel. Reusability results confirmed that the prepared catalyst is able to be reemployed up to 11 times. Statistical analysis has been performed using a Central Composite Design to evaluate the contribution and performance of the parameters on biodiesel purity.     

Development of microsatellite markers and their utilization in genetic diversity analysis of cultivated and wild populations of the mud carp （ Cirrhina molitorella）

Microsatellite markers have been increasingly used in genetic studies on fishery species because of their high applicability in selective breeding programs.Here we reported the development of microsatellite markers and their utilization in mud carp(Cirrhina molitorella).An (CA)15 enriched library has been constructed for mud carp,using the magnetic beads enrichment procedure.Sequence analysis of 60randomly picked positive colonies indicate that 56 (93.3%) of the colonies contain microsatellites.Microsatellite polymorphism was as-sessed using 10 mud carp individuals,and 12 microsatellite loci turned out to be polymorphic.We utilized these loci to study the genetic diversity of a wild population (WM) and a cultured population (CM) of the mud carp.A total of 109 alleles were detected with an average of 9.08 alleles per locus.The mean value of the observed heterozygosity of WM and CM was 0.6361 and 0.6417,respectively,and sig-nificant decrease of genetic diversity in CM was not observed.The genetic distance between the two populations was 0.1546 and the value of Gsr was 0.0473.This showed that there existed a slight genetic differentiation between WM and CM.     

Comparison of different osmolalities and egg-yolk composition in processing media for the cryopreservation of red wolf (Canis rufus) sperm

Successful cryopreservation of sperm and the maintenance of a sperm-based genome resource bank have been identified as priorities for the recovery of the endangered red wolf (Canis rufus). The objectives were to improve sperm processing and to determine the relative timing of damage to red wolf sperm during freezing and thawing. Fresh ejaculates (n=37) from adult red wolves (n=15, aged 2-13 y) were collected via electroejaculation and subjected to cooling, freezing and thawing in four TRIS-egg-yolk extender treatments varying in osmolality ( approximately 305 mOsm versus approximately 350 mOsm) and egg-yolk composition (0.8 microm-filtered versus unfiltered). Ejaculates were evaluated for sperm percentage motility, forward progressive motion, and morphological characteristics immediately upon collection and following extension, cooling (prior to freezing) and thawing. Although no single treatment consistently produced superior results, sperm suspended in approximately 305 mOsm extenders exhibited slight losses in motility post-thawing (13 and 7%). Also, sperm suspended in approximately 350 mOsm extenders tended to have slower rates of decline in motility in vitro post-thawing than those stored in approximately 305 mOsm extenders (P=0.55). Finally, extenders incorporating unfiltered egg yolk exhibited a slightly larger ratio of absent to partial acrosomes than did sperm frozen in extenders prepared with clarified egg yolk. For approximately 350 mOsm extenders, most motility loss occurred during the cooling rather than freezing and thawing. In conclusion, these data contribute to knowledge regarding cryopreservation of red wolf sperm.     

The use of the gonadotropin-releasing hormone analog deslorelin for short-term contraception in red pandas (Ailurus fulgens)

Red pandas (Ailurus fulgens) are threatened with extinction owing to habitat loss, exacerbated by their unique ecology and low fecundity. Regional breeding programs manage captive red panda populations. Recommendations not to breed may be made for various reasons, including genetic overrepresentation of certain individuals. No recommendations have been published on the use of contraception for red pandas. This article discusses the use of the GnRH analog deslorelin as a reversible method of contraception in both male and female pandas. The mean time from last contraception to conception was 3 years with a 4.6-mg deslorelin implant. The average dose of GnRH implant received was 1.09 mg/kg (range, 0.88–1.32). Males returned to breeding sooner than females. No reproductive side effects were noted with up to three consecutive annual GnRH implants.     

Isolation and characterization of 12 novel microsatellite loci for the red panda (Ailurus fulgens)

The information on dispersal patterns and mating systems of red pandas is quite important for the understanding of the genetic diversity and divergence of this species. And microsatellite marker is an ideal tool to analyze dispersal patterns and mating systems. Thus, we describe in this paper the isolation and characterization of 12 microsatellite loci in the red panda from genomic DNA-enriched libraries. These loci were highly polymorphic with numbers of alleles per locus in 24 individuals ranging from 2 to 14, observed heterozygosity from 0.143 to 0.864 and expected heterozygosity from 0.297 to 0.872. All loci except for RP6 locus followed Hardy–Weinberg expectations. No significant linkage association was found among all these loci. The 12 novel polymorphic microsatellite loci will be of use in studying dispersal patterns and mating systems of red pandas.     

Sucking and allosucking duration in farmed red deer (Cervus elaphus)

Sucking duration in ungulates does not only mean milk transfer, but is also associated with maternal care in general. It seems to be a reflection of offspring demand rather than solely milk transfer rate. Thus, the objective of this study was to discriminate between sucking and allosucking (i.e. sucking non-maternal hind) behaviour in red deer according to the sucking duration.We hypothesized that: (1) calves should suck longer from their mothers than allosuck from non-maternal hinds; (2) sucking duration of calves frequently nursed by a particular non-maternal hind should be longer than that of calves occasionally allonursed; (3) sucking duration should be longer for bouts including one calf than two or more calves sucking simultaneously; (4) male calves should suck and allosuck longer than female calves; and (5) primiparous hinds should nurse and allonurse longer than multiparous hinds. We observed sucking behaviour of 25 hinds and their 38 calves (from birth until the youngest calf reached one month of age) in two seasons. We recorded 1730 sucking bouts, of which 11.62% in the first season and 4.37% in the second season were non-filial. The duration of filial sucking was significantly longer than non-filial sucking. A large individual variance in the incidence of non-filial sucking in both the calves and hinds was found. Therefore, the non-filial hind–calf pairs were categorized in two clusters according to the frequency of nursing non-filial calves for one hind in relationship to all nursing events for this hind by a cluster analysis (PROC CLUSTER, SAS). We used a general linear mixed model, GLMM (PROC MIXED, SAS) to test the influence of hind relationship to the nursed calf (filial, frequently allosucking non-filial, or occasionally allosucking non-filial pair). Sucking duration of occasionally allosucking non-filial calves was only marginally different from that of filial calves. There was no difference between the two groups of non-filial calves. Multiple sucking bouts were shorter than those with one calf. Male calves sucked longer than female calves; however, the greatest difference was recorded between frequently allosucking non-filial pairs of both sexes. Frequently allosucking non-filial males sucked the longest and differently from occasionally allosucking non-filial males. Frequently allosucking non-filial females sucked the shortest and differently from filial calves of both sexes. It is more likely that allosucking seems to be more important for male rather than female calves.Therefore, it is concluded that allosucking calves differ in their sucking behaviour and two types of allosuckers (frequent and occasional) should be taken into account when analyzing allosuckling behaviour.     

Glutathione reductase activity and pyridoxine (pyridoxamine) phosphate oxidase activity in the red cell

The red-cell enzymes, glutathione reductase (FAD-dependent) and pyridoxine (pyridoxamine) phosphate oxidase (FMN-dependent), were studied in control subjects. The wide range the glutathione reductase activity correlated inversely with the percentage stimulation by FAD added in vitro, and with pyridoxine (pyridoxamine) phosphate oxidase activity. Both enzymes were stimulated after ingestion of riboflavin. The results support the suggestion that the rate of metabolism of riboflavin in the red cell controls the activity of both enzymes, and the rate of red-cell metabolism of vitamin B-6.