Sampling plans for estimating pepper fruit damage levels by Oriental tobacco budworm,Helicoverpa assulta (Guenee), in hot pepper fields

Sequential sampling programs for the management of Oriental tobacco budworm, Helicoverpa assulta (Guenee), on red hot peppers were developed using the data of damaged pepper fruits by H. assulta. Taylor's power law indicated that the damaged pepper fruits were distributed randomly in hot pepper fields. A fixed-precision-level sequential sampling plan for classifying fruit damage density levels at a critical density of 2 damaged fruits per plant was developed to assist in decision making. The sequential classification sampling plan was evaluated using the operating characteristic (OC) and the average sample size (ASN) curves. The OC and ASN curves indicated that this sampling plan was robust and properly classified the population density. A resampling simulation demonstrated that average actual sampling precision value at D = 0.25 was ≤ 0.25. With sequential sampling for classifying the damaged fruit levels in terms of a critical density, sample size was fixed to 18 plants. The fixed-precision-level sequential sampling plan developed in this study should greatly enhance the monitoring efficacy and provide practical solutions suitable for reliable decision-making process in the management of H. assulta.     

Towards an optimal sampling strategy for Alexandrium catenella (Dinophyceae) benthic resting cysts

The study proposes methodological developments to optimize sampling strategy of resting cysts of Alexandrium catenella to estimate their abundance with a predefined error. This work also aims to provide information on spatial distribution of resting cysts in sediments. The distribution mode of A. catenella resting cysts related to the abundance variability was studied through sediment cores sampling on four different spatial scales and using Ludox CLX gradient density method. The quantification method underestimates by a factor of 2 the resting cysts abundance in one gram of sediment. Application of Taylor's power law allowed us to define a compromise between sampling effort and abundance estimation error. In the case of A. catenella resting cysts from Thau lagoon, the optimal sampling strategy consists of sampling 10 stations on a surface of 2 km² for a given coefficient of variability (C) of 15%, sampling 3 sediment cores at each station (C = 30%) and counting only one replicate by core (C = 18%). Results related to the application of Taylor's power law are closely dependent on resting cyst density and aggregation in a given sediment. In our area, A. catenella resting cysts are mainly observed in the upper 3 cm of sediment. Horizontally, their heterogeneity is lower on 10 cm² surface and tends to stabilize itself beyond a surface of 10 m². Each author has to carry out this pre-sampling effort for his own resting cysts-forming species, in his own area, in order to increase accuracy of resting cyst mapping.     

Modifying orchard planting sites conserves entomopathogenic nematodes,reduces weevil herbivory and increases citrus tree growth,survival and fruit yield

In Florida, a root weevil pest of citrus, Diaprepes abbreviatus, is more damaging and attains higher population density in some orchards on fine textured, poorly drained “flatwoods” soils than in those on the deep, coarse sandy soils of the central ridge. Previous research revealed that sentinel weevil larvae were killed by indigenous entomopathogenic nematodes (EPNs) at significantly higher rates in an orchard on the central ridge, compared to one in the flatwoods. We hypothesized that filling tree planting holes in a flatwoods orchard with sandy soil from the central ridge would provide a more suitable habitat for EPNs, thereby reducing weevil numbers and root herbivory. Fifty trees were planted in oversized planting holes filled with coarse sand and 50 trees were planted in native soil in a split plot design where whole plots were species of introduced EPNs and split plots were soil type. Each of Steinernema diaprepesi, Steinernema riobrave, Heterorhabditis indica, Heterorhabditis zealandica, or no EPNs were introduced into the rhizospheres in 10 plots of each soil type. During four years, EPN numbers in soil samples and the relative abundance of seven species of nematophagous fungi associated with nematodes were measured three times using real-time PCR. The efficacy of EPNs against sentinel weevil larvae was also measured three times by burying caged weevils in situ. EPN species richness (P = 0.001) and diversity (P = 0.01) were always higher in sand than native soil. Soil type had no effect on numbers of EPNs in samples, but EPNs were detected more frequently (P = 0.01) in plots of sandy soil than native soil in 2011. Two nematophagous fungi species, Paecilomyces lilacinus and Catenaria sp. were significantly more abundant in nematode samples from sandy soil on all three sampling dates. Efficacy of EPNs against weevil larvae was greater in sandy soil inoculated with S. diaprepesi (P = 0.03) in June 2010 and in all treatments in sandy soil in May 2011 (P = 0.03). Sixty-eight percent more adult weevils (P = 0.01) were trapped emerging from native soil during two years than from sandy soil. By May 2011, the cumulative number of weevils emerging from each plot was inversely related (P = 0.01) to the numbers of EPNs detected in plots and to EPN efficacy against sentinels. Three trees in sandy soil died as a result of root herbivory compared to 21 trees in native soil. Surviving trees in sandy soil had trunk diameters that were 60% larger (P = 0.001) and produced 85% more fruit (P = 0.001) than those in native soil. Although it is not possible to characterize all of the mechanisms by which the two soil treatments affected weevils and trees, substitution of sand for native soil was an effective means of conserving EPNs and shows promise as a cultural practice to manage D. abbreviatus in flatwoods citrus orchards with a history of weevil damage to trees.     

Sampling of Bemisia tabaci adults using a pre-programmed autonomous pest control robot

An autonomous robot (Cabbot) was built to control insect pests in a plastic greenhouse and utilized to sample adult whiteflies (Bemisia tabaci) on paprika plants (Capsicum annum var. angulosum). To accomplish this, a sampling device consisting of an air compressor for pest agitation, a sticky trap (100 mm × 150 mm) and an image processing system for pest identification were installed on the Cabbot. The sampling precision of the Cabbot (D = 0.16) was higher than that of the sticky trap (D = 0.19) when sampling of adult white flies was conducted in caged pots. The Cabbot could also collect a substantial number of individuals within a short duration (3 minutes). The collection efficiency (i.e., percent of samples collected at least n (1 in this case) individuals to the total sample number) of insects by the Cabbot was markedly high at low population size, showing approximately 30% when the population size was ≤ 16 individuals per plant. The sampling precision and collection efficiency suggested that the Cabbot is effective in local, short-term sampling and could be used for early warning of pest occurrences.     

Variation of soil nutrients and particle size under different vegetation types in the Yellow River Delta

The Yellow River Delta wetland, located at the southern coast of Bohai Gulf, provides important ecosystem services such as flood control, water purification, biodiversity conservation, nutrient removal and carbon sequestration, shoreline stabilization, tourism attraction and wetland products maintains in the Yellow River Delta. This study assessed how agricultural activities in a reclamation wetland changed soil pH, soil electric conductivity, soil nutrient and soil particle size as compared to natural vegetation by using a combination of field experiments and lab analysis. The vegetation type included adjacent alfalfa field (Medicago sativa), cotton field (Gossypium spp.), Chinese tamarisk shrub (Tamarix chinensis), and reed marsh (Phragmites sage). The results indicated that the soil pH was higher (pH > 8) in alfalfa field and cotton field, and alfalfa field and reed marsh had significant function in reducing soil salt content, soil electric conductivity of alfalfa field at 0–30 cm were 140.38 ± 14.36, 114.48 ± 14.36, 125.30 ± 11.37 μs/cm. The effect of different vegetation types on soil nutrient was significant (P < 0.05). Soil organic matter at 0–10 cm in Chinese tamarisk shrub and reed marsh was 21.66 ± 3.82 g/kg and 16.51 ± 4.60 g/kg, which was higher than that of alfalfa field (10.47 ± 2.36 g/kg) and cotton field (9.82 ± 1.27 g/kg), but soil total nitrogen content in alfalfa field was the highest, which is significantly higher than that of cotton field, Chinese tamarisk shrub and reed marsh(P < 0.05), the content of soil total nitrogen at 0–10 cm and 10–20 cm was 7.67 ± 0.38 g/kg and 5.97 ± 0.51 g/kg, respectively, while the content of available P and available K was reversed. The difference of soil particle size between layers was not significant (P > 0.05), the sand content of Chinese tamarisk shrub soils in 0–10 cm was the highest, the next was alfalfa field and cotton field, and the content of silt and clay in reed marsh was higher than the others. The correlation and significant degree between soil particle size and soil nutrient was related with vegetation types, and soil organic matter was significantly positively correlated with soil silt and clay content on the alfalfa field. The results demonstrated that wetland cultivation was one of the most important factors influencing on the nutrient fate and reserves in soil, which could lead to rapid nutrient release and slow restoration through abandon cultivation. Consequently, compared with cotton field, alfalfa field is more favorable to sustainable management of wetland cultivation in the Yellow River Delta. It should be considered in wetland restoration projects planning.     

Associations of soil iron with citrus tree decline and variability of sand,soil water,pH, magnesium and Diaprepes abbreviatus root weevil: Two-site study

The hypothesis of associations of environmental soil heterogeneity with citrus tree decline and Diaprepes abbreviatus (L.) root weevil variability was tested in two flatwoods fields of ‘Hamlin’ orange trees (Citrus sinensis (L.) Osb.). Studies were conducted on a loamy, poorly drained Mollisol in Osceola County, central Florida in 2002, and on a sandy, poorly drained Spodosol in DeSoto County, south-west Florida during 2001–2003. Adult weevils were monitored using 50 Tedders traps arranged in a 34 m × 25 m grid at the Osceola site, and using 100 identical traps in a 30 m × 15 m grid at the DeSoto site. Soil water content (SWC), texture, pH, Ca, Mg, Fe, Cu and other nutrients were measured at each trap. Soil was strongly acidic (pH 4.9 ± 0.4) at the Osceola site but near neutral (pH 6.6 ± 0.4) at the DeSoto site. The Mehlich-I extractable soil Mg and Ca were correlated to soil pH and SWC in both soils, and extractable Fe was related to pH, SWC and Mg in the Spodosol (0.30 < R² < 0.65, P < 0.01). The weevil density was high in areas low in soil Mg and Ca in the acidic Mollisol, but high in areas with high soil pH, and Mg and low sand content in the near neutral Spodosol (P < 0.05). Tree decline was associated with soil Fe concentrations >40 mg kg⁻¹ in the Mollisol (P < 0.01). Weevil density was low at a soil pH between 5.7 and 6.2. The range of spatial dependence of weevil population, soil pH, SWC, Fe, Mg and sand varied between 60 and 100 m in the Mollisol and the Spodosol. Soil-weevil-tree simple and multivariate linear models were established to put into practices for predicting and controlling the weevil population and tree decline in the future. Differences in site characteristics suggested the need for site-specific weevil and citrus tree management.     

Alexandrium pacificum Litaker sp. nov (Group IV): Resting cyst distribution and toxin profile of vegetative cells in Bizerte Lagoon (Tunisia,Southern Mediterranean Sea)

A high spatial resolution sampling of Alexandrium pacificum cysts, along with sediment characteristics (% H₂O, % organic matter (OM), granulometry), vegetative cell abundance and environmental factors were investigated at 123 study stations in Bizerte Lagoon (Tunisia). Morphological examination and ribotyping of cells obtained from a culture called ABZ1 obtained from a cyst isolated in lagoon sediment confirmed that the species was A. pacificum. The toxin profile from the ABZ1 culture harvested during exponential growth phase was simple and composed of the N-sulfocarbamoyl toxins C1 (9.82 pg toxin cell⁻¹), the GTX6 (3.26 pg toxin cell⁻¹) and the carbamoyl toxin Neo-STX (0.38 pg toxin cell⁻¹). The latter represented only 2.8% of the total toxins in this strain.High abundance of A. pacificum cysts correlated with enhanced percentages of water and organic matter in the sediment. In addition, sediment fractions of less than 63 μm were examined as a favorable potential seedbed for initiation of future blooms and outbreaks of A. pacificum in the lagoon. A significant difference in the cyst distribution pattern was recorded among the lagoon's different zones, with the higher cyst abundance occurring in the inner waters. Also, no correlation due to the specific hydrodynamics of the lagoon was observed in the spatial distribution of A. pacificum cysts and vegetative cells.     

In situ degradability of seven plant protein supplements in heifers fed high concentrate diets with different forage to concentrate ratio

Four Holstein heifers (297.5 ± 27.7 kg BW) fed high concentrate diets were used in a crossover experiment in order to characterize the rumen fermentation pattern, and to estimate by the in situ method rumen degradation kinetics of alfalfa hay and seven plant protein supplements: solvent-extracted soybean meal, solvent-extracted sunflower meal, peas (Pisum sativum L.), lupin seeds (Lupinus sp.), broadbean (Vicia faba L.), horsebean (Vicia faba L. var equina) and vetch (Vicia sativa L.), in high concentrate diets with different forage to concentrate ratio. Heifers were fitted with a ruminal cannula. The experiment was performed in two 30-day periods, 15 days of diet adaptation and 15 days of sampling. At each period, heifers were offered one of two total mixed rations (12:88 versus 30:70 forage to concentrate ratio), two heifers per diet, on ad libitum basis. After the first period, heifers switched treatments. Intake of dry matter (DM), organic matter, crude protein and neutral detergent fibre (NDF), expressed as kg/day, did not differ between treatments, but DM intake, expressed as g/kg metabolic body weight (BW), was higher in the 12:88 diet. Average rumen pH was 6.0 in both diets, and the time pH was below 5.8, which is considered as a critical threshold for fibre degradation, was the same for both treatments (10.4 ± 1.6 h). Average ammonia nitrogen and volatile fatty acid (VFA) concentrations did not differ between treatments and individual VFA proportions were typical of high concentrate diets. Average effective degradability of DM (0.62 ± 0.02) and NDF (0.25 ± 0.03) of alfalfa hay were low and no differences were detected between treatments. The same extent of NDF degradation, together with the same proportions of VFA would indicate that both diets had the same fibrolytic activity. Forage to concentrate ratio did not affect rumen nitrogen degradability of any protein supplements incubated in situ. Corrected effective degradability for small particle losses of sunflower meal (0.78) was higher than legume seeds, which were not statistically different between each other and ranged from 0.63 to 0.66. Soybean meal had the lowest degradability value (0.61). These nitrogen degradation values must be considered more valid for beef cattle formulation of high concentrate diets than data obtained with forage diets.     

Validation of an electrospray ionization LC/MS/MS method for quantitative analysis of vincristine in human plasma samples

Vincristine is a natural vinca alkaloid widely used in paediatric cancer treatment. Vincristine pharmacokinetics has been already studied, but few data are available in paediatric populations. A sensitive and specific liquid chromatography–tandem mass spectrometry (LC/MS/MS) method was developed for the quantification of vincristine in plasma in order to investigate pharmacokinetics in a paediatric population. Two hundred microliters of plasma was added to vinblastine, used as internal standard. Chromatographic separation was achieved on a C8 HPLC column (Phenomenex Luna 50 mm × 2.0 mm, 3.0 μm) with a mobile phase gradient at a flow rate of 0.2 ml/min. Quantification was performed using the transition of 825.4 → 765.4 (m/z) for vincristine and 811.4 → 751.4 (m/z) for vinblastine. Chromatographic separation was achieved in 8 min. The limit of quantification was 0.25 ng/ml with a precision of 10.2% and an accuracy of 99.6%. The calibration curve was linear up to 50.0 ng/ml. Intra-day precision and accuracy ranged from 6.3% to 10% and from 91.9% to 100.8%, respectively. Inter-assay precision and accuracy ranged from 3.8% to 9.7% and from 93.5% to 100.5%, respectively. No significant matrix effect was observed for vincristine. A rapid, specific and sensitive LC/MS/MS method for quantification of vincristine in human plasma was developed and is now successfully applied for pharmacokinetic studies in paediatric patients.     

Distribution patterns of and sampling plans forTetranychus urticae Koch (Acarina: Tetranychidae) on roses

I sampled Tetranychus urticae Koch (Acarina: Tetranychidae) regularly from four rose gardens in Kyoto and Nara Prefectures in 1988–1990. When mite density was low, T. urticae showed an uneven vertical distribution, being more abundant in the lower third and absent from the upper third of the plants. Mite density was less variable within than between plants, suggesting that a sampling plan which includes more leaves from different plants rather than from different levels of a plant is favourable. Spatial distribution of T. urticae was nonrandom and followed the negative binomial distribution. In addition, both the Taylor's power law and the Iwao's patchiness regression described the distribution well. An empty-sample method for estimating mite density from the proportion of empty sampling units was developed. Sampling plans for determining the sample size required to reach a predetermined precision level, based on this method and by directly counting the mites, were designed. The counting method was more accurate than the empty-sample method. However, when the time factor was taken into account, the latter was more favourable, because it was faster than the former at a density range of 1.5–300 mites leaf⁻¹, which was most commonly encountered in the field.     

The historical dendroarchaeology of two log structures at the Wynnewood State Historic Site,Castalian Springs,Tennessee, USA

Tree rings of eastern red cedar (Juniperus virginiana L.) were examined from cores extracted from two log cabins located at the Wynnewood State Historic Site in Castalian Springs, Sumner County, Tennessee. One cabin was reportedly built by the first explorer in the area, Isaac Bledsoe, sometime between 1772 and his death in 1793. The second cabin was known as Spencer's Cabin after the first settler of the region, Thomas Sharp Spencer, who lived in the immediate vicinity from 1776 to 1779. The goal of this research was to determine the probable construction year(s) for both cabins and determine whether Bledsoe and Spencer did indeed build these structures. Forty-one cores were extracted from Bledsoe's Cabin, and 30 were used for crossdating and building a floating chronology using COFECHA. The Bledsoe's Cabin chronology was then statistically and graphically crossdated using the eastern red cedar reference tree-ring chronology (ITRDB #TN031) from Norris Dam, Tennessee. We found a statistically significant correlation (r = 0.42, t = 4.18, n = 85, p < 0.0001) between the Bledsoe's Cabin chronology and the reference chronology, anchoring the chronology between 1720 and 1804, with nearly all cores indicating tree harvesting between February and April 1805. Twenty-two cores were extracted from Spencer's Cabin, and 17 were used to build a floating chronology for the cabin. Again, we found a statistically significant correlation (r = 0.44, t = 4.85, n = 100, p < 0.00001) with the reference chronology, which anchored the Spencer's Cabin chronology between 1726 and 1825. All trees appear to have been harvested between February and August in 1826. Therefore, neither structure was built by its historical namesake. No known historical documents suggest who the potential builders were, although the property was owned between ca. 1797 and 1829 by General James Winchester. He and his family, however, never resided on the Wynnewood property because Winchester had built a large multi-room structure in nearby Gallatin, Tennessee, by 1802.     

Assessment of spatial distribution and cysts resources of Artemia in late autumn in Dangxiong Co salt lake

Based on the vertically interval sampling in 25 sampling sites in Dangxiong Co salt lake in 2011, a preliminary investigation on population spatial distribution and cysts resources of Artemia in the lake has been conducted. The study achieves four new progresses. First, the average density of Artemia and Artemia cysts in the lake is 4.157 × 10³ ind. m⁻² and 8.069 × 10⁴ ind. m⁻², respectively. Among Artemia, the adults account for 60.31%; Second, different from other salt lakes in horizontal distribution, the Artemia population mainly distributes in the open water, only a little in the shallow water, and there is no distribution in the longshore area in the north part and the estuary region; Third, in vertical distribution, 44.24% of individuals intensively distribute in the upper water layer within 2.0 m, especially 0–0.2 m, where the average density of Artemia and cysts are maximum (129.488 ind. L⁻¹ and 5.728 ind. L⁻¹, respectively). A decrease of distribution density is accompanied by an increase of water depth basically, the percentage of Artemia and cysts decrease to 0.68% and 4.60%, respectively; Fourth, the cysts resources of 14.96 t in the lake are assessed using contour map. 66.35% of them distributed in 0.0–2.0 m water layer and 49.06% concentrate in the 18.21% areas of the central water; Fifth, quantity of cysts suitable for development in the lake is 2.399 ± 0.320 t, with an upper limit of 0.879 t. The study can provide a reference for the sustainable development and exploitation of Artemia in Dangxiong Co salt lake.     

Factors supporting harmful macroalgal blooms in flowing waters: A 2-year study in the Lower Ain River,France

A two-year study was conducted to explore summer development of macroalgae and their total phosphorus and nitrogen content at three stations in a broad and clear French carbonate river. Water discharge, temperature and insolation, each with a different time lag, as well as substrates and nutrients were examined in order to explain macroalgal biomass variability. Twenty-four macroalgae genera were recorded with Spirogyra, Cladophora, Vaucheria and Oedogonium as abundant. Through redundancy analysis the macroalgal community composition exhibited significant differences, between the sampling sites and also from one year to the next. Water discharge (time-lag = 5 days) and temperature (time-lag = 20 days) both significantly explained macroalgal biomass variability, highlighting differences in the time lag of the macroalgal community's ecological response to environmental changes. Spatial segregation was observed within the wide riverbed due to habitat variability, allowing co-occurrence in the development of ecologically different taxa within each sampling site. The high nitrate concentrations as compared with the particular low phosphorus concentrations led to especially high DIN/SRP ratios (248 ± 103, n = 18). The N/P ratios in algal tissues were high (25 ± 16, n = 26) and indicated P-limitation. The differences in DIN/SRP and N/P ratios suggest additional nutrient sources than open water such as groundwater inputs.     

Effect of Gynostemma Pentaphyllum Polysaccharide on boar spermatozoa quality following freezing–thawing

Gynostemma Pentaphyllum Polysaccharide (GPP) was added at concentrations of 0.25, 0.5, 1.0, 1.5 and 2.0 mg/ml to the extenders used to freeze boar semen and its effects on the quality of frozen–thawed sperm were assessed. The sperm motility was significantly higher in the extenders containing 0.25 and 0.5 mg/ml GPP, as compared to other groups (P < 0.05). The extender supplemented with 0.5 mg/ml GPP favored the highest intact membrane and intact acrosome percentages in comparison with other groups (P < 0.05), respectively. The mitochondrial activity was significantly higher at the concentrations of 0.25, 0.5 and 1.0 mg/ml GPP than that of other treatments, and the control group (P < 0.05). In biochemical assays, the extender supplemented with 0.25 and 0.5 mg/ml GPP significantly improved SOD levels, compared to other groups (P > 0.05). However, the extenders supplemented with GPP did not cause significant differences in levels of CAT and GSH-Px, compared to the control (P > 0.05). In summary, GPP exhibited a dose-related response and the lower concentration produced greater protective effect. According to the standard semen quality parameters and antioxidant activities measured in this study, the concentration of 0.5 mg/ml GPP caused a beneficial cryoprotective effects on the quality of frozen–thawed boar semen. It is proposed that an extender containing 0.5 mg/ml GPP could be used as cryoprotective medium of better efficiency.     

Primer caso de fungemia asociada a catéter por Candida nivariensis en la Península Ibérica

BackgroundIn recent years the incidence of candidemia caused by non-albicans Candida species has been increasing. Two cryptic species have been described within the Candida glabrata complex, Candida nivariensis and Candida bracarensis, which may be troublesome in laboratory identification and have lower susceptibility to fluconazole.AimsTo describe the first isolation of C. nivariensis in the Iberian Peninsula from a patient suffering from a catheter-related fungemia.Case reportAn 81-year-old man was hospitalized for surgical treatment of an intestinal fistula that was associated to a severe malnutrition. Cultures of the patient's central venous catheter tip and blood yielded white colonies in BD CHROMagar Candida^® medium, which could not be identified by conventional microbiological methods. Although intravenous fluconazole was administered, blood cultures continued being positive 5 days later. The MIC values of the isolate were as follows: 1 μg/ml for amphotericin B, 0.015 μg/ml for anidulafungin, 0.125 μg/ml for caspofungin, 0.015 μg/ml for micafungin, 4 μg/ml for fluconazole, 0.25 μg/ml for itraconazole, 0.25 μg/ml for posaconazole, and 0.03 μg/ml for voriconazole. Antifungal treatment was changed to intravenous caspofungin for 2 weeks. The intestinal fistula was surgically treated. There was no evidence of relapse during the following month, and the patient was discharged. The isolate was identified as C. nivariensis based on DNA sequencing of the ITS regions of rRNA.ConclusionsC. nivariensis should be regarded as an emerging pathogen which requires molecular methods for a definitive identification. Our patient was successfully treated with caspofungin.     

Chloroplast DNA variation and phylogeographic patterns in the Chinese endemic marsh herb Sagittaria potamogetifolia

We examined chloroplast DNA (cpDNA) atpB–rbcL intergenic spacer sequences variation within Sagittaria potamogetifolia, an endangered and endemic marsh herb in China. Sequence data were obtained from 54 individuals in six extant populations of the species. Sequences appeared to evolve neutral (Tajima's criterion D = −1.59826, 0.1 > P > 0.05 and Fu and Li's tests D* = −1.44484, P > 0.1; F* = −1.83446, P > 0.1). Eleven haplotypes were identified in S. potamogetifolia. A relatively high level of haplotype diversity (h = 0.0.699) and low level of nucleotide diversity (pi = 0.0035 ± 0.0020) were detected in S. potamogetifolia. Pairwise comparisons of F_st and Nm deduced from cpDNA variation suggested no significant genetic differentiation between populations of S. potamogetifolia excepted for the WY-1 population. Low genetic differentiation among populations and also among regions was consistently indicated by both hierarchical analyses of molecular variance (AMOVA) and the structure of a neighbor-joining tree. Lack of population differentiation between populations or between regions in cpDNA sequences may be due to effects of lower substitution rates or lineage sorting. In the minimum spanning network, all tip haplotypes except for the haplotype J were unique to a particular population, while the interior nodes except for the haplotype E were widespread (haplotype A). From nested clade analysis (NCA), the evolutionary events such as restricted gene flow with isolation by distance and allopatric fragmentation were inferred to responsible for the current distribution of S. potamogetifolia populations, as well as their genetic diversity.     

Liquid chromatography/tandem mass spectrometry method for the simultaneous determination of vardenafil and its major metabolite,N-desethylvardenafil,in human plasma: Application to a pharmacokinetic study

A rapid and sensitive LC–MS/MS method for the determination of vardenafil and its major metabolite, N-desethylvardenafil, in human plasma using sildenafil as an internal standard was developed and validated. The analytes were extracted from 0.25-mL aliquots of human plasma by liquid–liquid extraction, using 1 mL of ethyl acetate. Chromatographic separation was carried on a Luna C₁₈ column (50 mm × 2.0 mm, 3 μm) at 40 °C, with an isocratic mobile phase consisting of 10 mM ammonium acetate (pH 5.0) and acetonitrile (10:90, v/v), a flow rate of 0.2 mL/min, and a total run time of 2 min. Detection and quantification were performed using a mass spectrometer in the selected reaction-monitoring mode with positive electrospray ionization at m/z 489.1 → 151.2 for vardenafil, m/z 460.9 → 151.2 for N-desethylvardenafil, and m/z 475.3 → 100.1 for the internal standard (IS), respectively. This assay was linear over a concentration range of 0.5–200 ng/mL with a lower limit of quantification of 0.5 ng/mL for both vardenafil and N-desethylvardenafil. The coefficient of variation for the assay precision was <13.6%, and the accuracy was >93.1%. This method was successfully applied to a pharmacokinetic study after oral administration of vardenafil 20 mg tablet in Korean healthy male volunteers.     

A maze-lover's dream: Burrow architecture,natural history and habitat characteristics of Ansell's mole-rat (Fukomys anselli)

The Ansell's mole-rat (Fukomys anselli, Bathyergidae) is a small-sized social subterranean rodent whose distribution is confined to the Lusaka area in Zambia. It is an established model species for various laboratory studies, but until now the knowledge of its biology under natural conditions has been limited. Here, we provide the first comprehensive natural history and ecological data on a free living population from Miombo woodland. The Ansell's mole-rat lives in groups of up to 13 individuals (mean 9.7 ± 2.5; N = 9 complete, fully established family groups) with a single breeding pair per group. The family groups occupy very large and complex burrow systems consisting of 0.5–2.8 km of tunnels (mean = 1.2 ± 0.6 km, N = 15), which are densely branched and reticulated especially around nests. Burrow systems of neighbouring family groups were often interconnected by a freely passable tunnel, which has never before been documented in any African mole-rats. Mole-rat food density in the study area was relatively low (10.0 ± 8.4 m^?2) but its biomass was large (457 ± 889 g m^?2). This, together with a relatively workable soil (cone resistance = 529 ± 26 N cm^?2 at the end of June, with 10% soil humidity), indicates moderate ecological conditions, which disagrees with the aridity food-distribution hypothesis (AFDH) considering small body size and sociality in bathyergids adaptations to harsh environments.     

Mechanical properties of the patellar tendon in adults and children

It is not currently known how the mechanical properties of human tendons change with maturation in the two sexes. To address this, the stiffness and Young's modulus of the patellar tendon were measured in men, women, boys and girls (each group, n=10). Patellar tendon force (F_pt) was calculated from the measured joint moment during a ramped voluntary isometric knee extension contraction, the antagonist knee extensor muscle co-activation quantified from its electromyographical activity, and the patellar tendon moment arm measured from magnetic resonance images. Tendon elongation was imaged using the sagittal-plane ultrasound scans throughout the contraction. Tendon cross-sectional area was measured at rest from ultrasound scans in the transverse plane. Maximal F_pt and tendon elongation were (mean±SE) 5453±307 N and 5±0.5 mm for men, 3877±307 N and 4.9±0.6 mm for women, 2017±170 N and 6.2±0.5 mm for boys and 2169±182 N and 5.9±0.7 mm for girls. In all groups, tendon stiffness and Young's modulus were examined at the level that corresponded to the maximal 30% of the weakest participant's F_pt and stress, respectively; these were 925–1321 N and 11.5–16.5 MPa, respectively. Stiffness was 94% greater in men than boys and 84% greater in women than girls (p<0.01), with no differences between men and women, or boys and girls (men 1076±87 N/mm; women 1030±139 N/mm; boys 555±71 N/mm and girls 561.5±57.4 N/mm). Young's modulus was 99% greater in men than boys (p<0.01), and 66% greater in women than girls (p<0.05). There were no differences in modulus between men and women, or boys and girls (men 597±49 MPa; women 549±70 MPa; boys 255±42 MPa and girls 302±33 MPa). These findings indicate that the mechanical stiffness of tendon increases with maturation due to an increased Young's modulus and, in females due to a greater increase in tendon cross-sectional area than tendon length.     

The effect of wave exposure on the morphology of Ecklonia radiata

This study examined the consistency of the effect of wave exposure on the morphology of Ecklonia radiata, a small kelp, across a broad geographic range (>1100 km). Fifteen morphological characters were measured on individuals from sites of low (2.5 ± 0.8 S.E.) and high (12.2 ± 2.0 S.E.) wave exposure (Baardseth's index) within six locations in southwestern Australia. With the exception of lateral width (P = 0.0001), none of the morphological characters were consistently statistically different (P > 0.06) between high and low wave exposure and correlations with wave exposure were generally weak (r < 0.60). While 12 of the 15 characters were statistically different (P < 0.008) between sites of different exposure within at least one location, the direction of difference (significant or not) was opposite between some locations for all but one character (lateral width). ANOSIM was not able to separate thalli from high and low exposure when all locations were pooled (Clarke's R = 0.127), but within each location most sites showed better separation (0.126 < Clarke's R < 0.829). Despite the lack of statistical differences, trends suggested that E. radiata responds to exposure by having drag-reducing (small size, narrow laterals and blades, low spinosity) and strength-increasing (relatively large holdfast, thick stipe and thick blades and lamina) morphological traits, as observed for several other kelps in small-scale studies. We conclude that while wave exposure does have an effect on kelp morphology, the effect is not independent of other location-specific processes.