Progeny of the maize weevil,Sitophilus zeamais,is affected by parental exposure to clove and cinnamon essential oils

The interest in and utilization of botanical insecticides, particularly essential oils, has become increasingly relevant to the control of insect pests. However, the potential ecotoxicological risks or flaws (including sublethal effects on the targeted pest generation and its subsequent progeny) of this pest control tool have been neglected frequently. Here, we evaluated the effects of sublethal exposure to clove, Syzygium aromaticum (L.) Merrill & Perry (Myrtaceae), and cinnamon, Cinnamomum verum J. Presl (Lauraceae), essential oils on adult (F0) maize weevils, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and the physiology (e.g., body mass, respirometry, and grain consumption) and population dynamics (e.g., daily emergence and sex ratio) of their progeny. Longevities of the parents were negatively affected by the essential oils in a concentration‐dependent manner. Parental sublethal exposure to clove oil (0.17 μl cm^?2) accelerated offspring emergence but delayed the emergence of females compared to males. Parents that were sublethally exposed to clove (0.17 μl cm^?2) or cinnamon (0.35 μl cm^?2) essential oils produced heavier offspring. Parental sublethal exposure to cinnamon essential oil accelerated offspring emergence (at 0.70 μl cm^?2), delayed female emergence (at 0.17 μl cm^?2), and enhanced grain consumption (at 0.35 and 0.70 μl cm^?2) of the progeny. Thus, our findings indicate that sublethal exposure to clove and cinnamon essential oils is capable of promoting transgenerational effects in S. zeamais that can negatively impact the control efficacy of such products.     

A comparison of isoprene and monoterpene emission rates from the perennial bioenergy crops short‐rotation coppice willow and Miscanthus and the annual arable crops wheat and oilseed rape

Biogenic volatile organic compounds (BVOC) emissions from bioenergy crops may differ from those of conventional crops. We compared emission rates of isoprene and a number of monoterpenes from the lignocellulosic bioenergy crops short‐rotation coppice (SRC) willow and Miscanthus, with the conventional crops wheat and oilseed rape. BVOC emission rates were measured via dynamic vegetation enclosure and GC‐MS analysis approximately monthly between April 2010 and August 2012 at a location in England and from SRC willow at two locations in Scotland. The largest BVOC emission rates were measured from willow in England and varied between years. Isoprene emission rates varied between μg g^?1 h^?1. Of the monoterpenes detected from willow, α‐pinene emission rates were highest (μg g^?1 h^?1), followed by μg g^?1 h^?1 for δ‐3‐carene, μg g^?1 h^?1 for β‐pinene and μg g^?1 h^?1 for limonene. BVOC emission rates measured in Scotland were much lower. Low emission rates of isoprene and α‐pinene were measured from Miscanthus in 2010 (μg g^?1 h^?1 and μg g^?1 h^?1, respectively) but were not detected in subsequent years. Emission rates from wheat of isoprene were negligible but relatively high for monoterpenes (μg g^?1 h^?1 and μg g^?1 h^?1 for α‐pinene and limonene, respectively). No significant emission rates of BVOCs were measured from oilseed rape. The measured emission rates followed a clear seasonal trend. Crude extrapolations based solely on data gathered here indicate that isoprene emissions from willow could correspond to 0.004–0.03% (UK) and 0.76–5.5% (Europe) of current global isoprene if 50% of all land potentially available for bioenergy crops is planted with willow.     

Chemical Composition,Antimicrobial and Cytotoxic Activity of Heracleum verticillatum Pančić and H. ternatum Velen. (Apiaceae) Essential Oils

In this work, the chemical composition, antimicrobial and cytotoxic activity of Heracleum verticillatum Pan?i? and H. ternatum Velen . root, leaf, and fruit essential oils were investigated. The composition was analyzed by GC and GC/MS. Heracleum verticillatum and H. ternatum root oils were dominated by monoterpenes, mostly β‐pinene (23.5% and 47.3%, respectively). Heracleum verticillatum leaf oil was characterized by monoterpenes, mainly limonene (20.3%), and sesquiterpenes, mostly (E)‐caryophyllene (19.1%), while H. ternatum leaf oil by the high percentage of phenylpropanoids, with (Z)‐isoelemicin (35.1%) being dominant constituent. Both fruit oils contained the majority of aliphatic esters, mostly octyl acetate (42.3% in H. verticillatum oil and 49.0% in H. ternatum oil). The antimicrobial activity of the oils was determined by microdilution method against eight bacterial and eight fungal strains. The strongest effect was exhibited by H. verticillatum root oil, particularly against Staphylococcus aureus, Salmonella typhimurium (MICs = 0.14 mg/ml, MBCs = 0.28 mg/ml), and Trichoderma viride (MIC = 0.05 mg/ml, MFC = 0.11 mg/ml). Cytotoxic effect was determined by MTT test against malignant HeLa, LS174, and A549 cells (IC₅₀ = 5.9 – 146.0 μg/ml), and against normal MRC‐5 cells (IC₅₀ > 120.1 μg/ml). The best effect was exhibited by H. verticillatum root oil on A549 cells (IC₅₀ = 5.9 μg/ml), and H. ternatum root oil against LS174 cells (IC₅₀ = 6.7 μg/ml).     

Uptake kinetics and storage capacity of dissolved inorganic phosphorus and corresponding N:P dynamics in Ulva lactuca (Chlorophyta)

Dissolved inorganic phosphorus (DIP ) is an essential macronutrient for maintaining metabolism and growth in autotrophs. Little is known about DIP uptake kinetics and internal P‐storage capacity in seaweeds, such as Ulva lactuca (Chlorophyta). Ulva lactuca is a promising candidate for biofiltration purposes and mass commercial cultivation. We exposed U. lactuca to a wide range of DIP concentrations (1–50 μmol · L^?1) and a nonlimiting concentration of dissolved inorganic nitrogen (DIN ; 5,000 μmol · L^?1) under fully controlled laboratory conditions in a “pulse‐and‐chase” assay over 10 d. Uptake kinetics were standardized per surface area of U. lactuca fronds. Two phases of responses to DIP ‐pulses were measured: (i) a surge uptake (V_S ) of 0.67 ± 0.10 μmol · cm^?2 · d^?1 and (ii) a steady state uptake (V_M ) of 0.07 ± 0.03 μmol · cm^?2 · d^?1. Mean internal storage capacity (ISC_P ) of 0.73 ± 0.13 μmol · cm^?2 was calculated for DIP . DIP uptake did not affect DIN uptake. Parameters of DIN uptake were also calculated: V_S  = 12.54 ± 1.90 μmol · cm^?2 · d^?1, V_M  = 2.26 ± 0.86 μmol · cm^?2 · d^?1, and ISC_N  = 22.90 ± 6.99 μmol · cm^?2. Combining ISC and V_M values of P and N, nutrient storage capacity of U. lactuca was estimated to be sufficient for ~10 d. Both P and N storage capacities were filled within 2 d when exposed to saturating nutrient concentrations, and uptake rates declined thereafter at 90% for DIP and at 80% for DIN . Our results contribute to understanding the ecological aspects of nutrient uptake kinetics in U. lactuca and quantitatively evaluating its potential for bioremediation and/or biomass production for food, feed, and energy.     

Isolation of the New Antiplasmodial Butanolide,Malleastrumolide A,from Malleastrum sp. (Meliaceae) from Madagascar

An extract of Malleastrum sp. (Meliaceae) collected in Madagascar by the Madagascar International Cooperative Biodiversity Group was found to have antimalarial activity, with an IC ₅₀ value between 2.5 and 5 μg ml^?1. After purification by liquid‐liquid partition, chromatography on a Diaion open column, C ₁₈ SPE and C ₁₈ reversed phase HPLC , the new butanolide, malleastrumolide A, was isolated. The structure of malleastrumolide A was determined by mass spectrometry, NMR , and ECD . The double bond position was determined by cross‐metathesis and mass spectrometry. The compound has antiproliferative activity against the A2780 ovarian cancer cell line with an IC ₅₀ value of 17.4 μm and antiplasmodial activity against the drug‐resistant Dd2 strain of Plasmodium falciparum with an IC ₅₀ value of 2.74 μm .     

Understanding the stoichiometric limitation of herbivore growth: the importance of feeding and assimilation flexibilities

Ecological stoichiometry suggests that herbivore growth is limited by phosphorus when this element in the diet is < 8.6 μg P mg C^?1 (C : P atomic ratio > 300). However, in nature, it is not necessarily related to the relative phosphorus content in diets. This may be the result of complex feeding and assimilation responses to diets. We examined these possibilities using herbivorous plankton fed mono‐specific and mixed algae varying in phosphorus content of 1.6 to 8.1 μg P mg C^?1. The herbivores showed a 10‐fold growth rate difference among the diets. Growth rates related poorly with phosphorus content in the diets (r² = 0.07), better with P ingestion rate (r² = 0.41) and best with phosphorus assimilation rate (r² = 0.69). Inclusion of assimilation rates for carbon and fatty acids increased 7% of the explained growth variance. These results indicate that the feeding and assimilation flexibilities play pivotal roles in acquiring a deficient element and in regulating growth rate.     

Invasive submerged freshwater macrophytes are more plastic in their response to light intensity than to the availability of free CO2 in air‐equilibrated water

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Environmental costs and benefits of growing Miscanthus for bioenergy in the UK

Planting the perennial biomass crop Miscanthus in the UK could offset 2–13 Mt oil eq. yr^?1, contributing up to 10% of current energy use. Policymakers need assurance that upscaling Miscanthus production can be performed sustainably without negatively impacting essential food production or the wider environment. This study reviews a large body of Miscanthus relevant literature into concise summary statements. Perennial Miscanthus has energy output/input ratios 10 times higher (47.3 ± 2.2) than annual crops used for energy (4.7 ± 0.2 to 5.5 ± 0.2), and the total carbon cost of energy production (1.12 g CO₂‐C eq. MJ^?1) is 20–30 times lower than fossil fuels. Planting on former arable land generally increases soil organic carbon (SOC) with Miscanthus sequestering 0.7–2.2 Mg C4‐C ha^?1 yr^?1. Cultivation on grassland can cause a disturbance loss of SOC which is likely to be recovered during the lifetime of the crop and is potentially mitigated by fossil fuel offset. N₂O emissions can be five times lower under unfertilized Miscanthus than annual crops and up to 100 times lower than intensive pasture. Nitrogen fertilizer is generally unnecessary except in low fertility soils. Herbicide is essential during the establishment years after which natural weed suppression by shading is sufficient. Pesticides are unnecessary. Water‐use efficiency is high (e.g. 5.5–9.2 g aerial DM (kg H₂O)^?1, but high biomass productivity means increased water demand compared to cereal crops. The perennial nature and belowground biomass improves soil structure, increases water‐holding capacity (up by 100–150 mm), and reduces run‐off and erosion. Overwinter ripening increases landscape structural resources for wildlife. Reduced management intensity promotes earthworm diversity and abundance although poor litter palatability may reduce individual biomass. Chemical leaching into field boundaries is lower than comparable agriculture, improving soil and water habitat quality.     

Strain variability in fatty acid composition of Chattonella marina (Raphidophyceae) and its relation to differing ichthyotoxicity toward rainbow trout gill cells

Lipid profiles of three strains (Mexico, Australia, Japan) of Chattonella marina (Subrahmanyan) Hara et Chihara were studied under defined growth (phosphate, light, and growth phase) and harvest (intact and ruptured cells) conditions. Triacylglycerol levels were always <2%, sterols <7%, free fatty acids varied between 2 and 33%, and polar lipids were the most abundant lipid class (>51% of total lipids). The major fatty acids in C. marina were palmitic (16:0), eicosapentaenoic (EPA, 20:5ω3), octadecatetraenoic (18:4ω3), myristic (14:0), and palmitoleic (16:1ω7c) acids. Higher levels of EPA were found in ruptured cells (21.4–29.4%) compared to intact cells (8.5–25.3%). In general, Japanese N‐118 C. marina was the highest producer of EPA (14.3–29.4%), and Mexican CMCV‐1 the lowest producer (7.9–27.1%). Algal cultures, free fatty acids from C. marina, and the two aldehydes 2E,4E‐decadienal and 2E,4E‐heptadienal (suspected fatty acid‐derived products) were tested against the rainbow trout fish gill cell line RTgill‐W1. The configuration of fatty acids plays an important role in ichthyotoxicity. Free fatty acid fractions, obtained by base saponification of total lipids from C. marina showed a potent toxicity toward gill cells (median lethal concentration, LC₅₀ (at 1 h) of 0.44 μg · mL^?1 in light conditions, with a complete loss of viability at >3.2 μg · mL^?1). Live cultures of Mexican C. marina were less toxic than Japanese and Australian strains. This difference could be related to differing EPA content, superoxide anion production, and cell fragility. The aldehydes 2E,4E‐decadienal and 2E,4E‐heptadienal also showed high impact on gill cell viability, with LC₅₀ (at 1 h) of 0.34 and 0.36 μg · mL^?1, respectively. Superoxide anion production was highest in Australian strain CMPL01, followed by Japanese N‐118 and Mexican CMCV‐1 strains. Ruptured cells showed higher production of superoxide anion compared to intact cells (e.g., 19 vs. 9.5 pmol · cell^?1 · hr^?1 for CMPL01, respectively). Our results indicate that C. marina is more ichthyotoxic after cell disruption and when switching from dark to light conditions, possibly associated with a higher production of superoxide anion and EPA, which may be quickly oxidized to produce more toxic derivates, such as aldehydes.     

Development and evaluation of ForestGrowth‐SRC a process‐based model for short rotation coppice yield and spatial supply reveals poplar uses water more efficiently than willow

Woody biomass produced from short rotation coppice (SRC) poplar (Populus spp.) and willow (Salix spp.) is a bioenergy feedstock that can be grown widely across temperate landscapes and its use is likely to increase in future. Process‐based models are therefore required to predict current and future yield potential that are spatially resolved and can consider new genotypes and climates that will influence future yield. The development of a process‐based model for SRC poplar and willow, ForestGrowth‐SRC, is described and the ability of the model to predict SRC yield and water use efficiency (WUE) was evaluated. ForestGrowth‐SRC was parameterized from a process‐based model, ForestGrowth for high forest. The new model predicted annual above ground yield well for poplar (r² = 0.91, RMSE = 1.46 ODT ha^?1 yr^?1) and willow (r² = 0.85, RMSE = 1.53 ODT ha^?1 yr^?1), when compared with measured data from seven sites in contrasting climatic zones across the United Kingdom. Average modelled yields for poplar and willow were 10.3 and 9.0 ODT ha^?1 yr^?1, respectively, and interestingly, the model predicted a higher WUE for poplar than for willow: 9.5 and 5.5 g kg^?1 respectively. Using regional mapped climate and soil inputs, modelled and measured yields for willow compared well (r² = 0.58, RMSE = 1.27 ODT ha^?1 yr^?1), providing the first UK map of SRC yield, from a process‐based model. We suggest that the model can be used for predicting current and future SRC yields at a regional scale, highlighting important species and genotype choices with respect to water use efficiency and yield potential.     

Pharmacokinetics of oxytetracycline in yellow catfish [Pelteobagrus fulvidraco (Richardson, 1846)] with a single and multiple‐dose oral administration

A pharmacokinetic study of oxytetracycline (OTC) following a single (100 mg kg^?1) or a multi‐dose (100 mg kg^?1 for 5 days) oral administration was carried out in yellow catfish, Pelteobagrus fulvidraco. After oral administration at 25°C, a one‐compartment model was developed. The absorption half‐life (t_1/2(ka)) was 3.92, 1.44, 2.75, and 3.34 h in plasma, muscle, liver, and kidney after the single dose, and 0.35, 0.22, 0.42, 0.32 h after the multi‐dose, respectively. The order of peak concentration (C_max) was liver > kidney > plasma > muscle, at 3.48 μg g^?1, 2.90 μg g^?1, 1.46 μg ml^?1, and 1.39 μg g^?1 after the single dose, and 14.02 μg g^?1, 8.51 μg g^?1, 4.17 μg ml^?1, and 3.84 μg g^?1 after the multi‐dose, respectively. The elimination half‐lives (t_1/2(ke)) of OTC in plasma, muscle, liver, and kidney were calculated to be 7.64, 26.29, 19.08, and 10.61 h after the single dose, and 47.54, 70.99, 49.87, and 47.73 h after the multi‐dose, respectively. The results suggest that OTC was absorbed faster after the multi‐dose than after the single dose, suggesting that OTC could be more efficacious after the multi‐dose and more effective in the control bacterial diseases in aquaculture, with the side effects of longer withdrawal periods.     

广藿香叶盘法高效再生体系主要影响因素的研究

广藿香是重要的芳香药用植物,利用基因工程技术对广藿香进行品种改良,需要建立一个高效的广藿香植株再生体系。该研究以广藿香无菌苗叶片为材料,将叶盘外植体分别置于不同条件下培养,观察、统计其再生植株的数量及生长状况。通过研究15~50 d的苗龄、第2~4节上的叶及培养基中2,4-D、NAA、BA和KT的浓度和配比等因素对广藿香叶盘再生植株的影响,在此基础上优化培养条件,建立广藿香高效再生体系。结果表明:广藿香无菌苗的苗龄、叶片在茎上的着生位置以及培养基中的植物生长调节物质浓度和配比都对广藿香的植株再生有显著影响;优化培养条件为以培养30 d的广藿香无菌苗顶芽下第2对展开叶片切割的叶盘为外植体,在含0.1 mg·L-1NAA和0.5 mg·L-1BA的MS培养基中培养28 d,叶盘的不定芽发生频率达到100%,单个叶盘的平均再生芽数为96.5个,经生根培养及温室炼苗,再生植株的移栽成活率达到96%。广藿香叶盘植株再生体系的建立为其基因转化研究及优良品种的快速繁育奠定了基础。     

Variability in iodine in temperate seaweeds and iodine accumulation kinetics of Fucus vesiculosus and Laminaria digitata (Phaeophyceae,Ochrophyta)

The biogeochemistry of iodine in temperate coastal ecosystems is largely mediated by macroalgae, which act as a major biological sink and source of iodine. Their capacity to accumulate, retain and release iodine has been associated with abiotic and biotic stressors, but quantitative information is limited. We evaluated the seasonal iodine retention capacity of eleven macroalgal species belonging to different systematic groups, collected from two sites in Ireland. Iodine accumulation and retention were then further quantified in Fucus vesiculosus and Laminaria digitata in relation to I^? concentrations in seawater and temperature. In general, iodine contents were ~10¹–10² μmol · (g dw)^?1 for Laminariales, 10⁰–10¹ μmol · (g dw)^?1 for Fucales, 10^?1–10⁰ μmol · (g dw)^?1 for Rhodophyta, and 10^?1 μmol · (g dw)^?1 for Chlorophyta. Typically, algal iodine contents were above average in winter and below average in summer. Iodine accumulation in F. vesiculosus and L. digitata depended on I^? availability and followed the Michaelis‐Menten kinetic. The ratio of maximum accumulation rate to half accumulation coefficient (ρ_max: K _t) was 2.4 times higher for F. vesiculosus than for L. digitata , suggesting that F. vesiculosus was more efficient in iodine accumulation. Both species exhibited a temperature‐dependent net loss of iodine, and only an exposure to sufficient external I^? concentrations compensated for this loss. This study revealed that both environmental (e.g., I^? in seawater, temperature) and organismal (e.g., the status of the iodine storage pool) variables determine retention and variability in iodine in temperate seaweeds.     

Soil respiration is stimulated by elevated CO2 and reduced by summer drought: three years of measurements in a multifactor ecosystem manipulation experiment in a temperate heathland (CLIMAITE)

This study investigated the impact of predicted future climatic and atmospheric conditions on soil respiration (R_S) in a Danish Calluna‐Deschampsia‐heathland. A fully factorial in situ experiment with treatments of elevated atmospheric CO₂ (+130 ppm), raised soil temperature (+0.4 °C) and extended summer drought (5–8% precipitation exclusion) was established in 2005. The average R_S, observed in the control over 3 years of measurements (1.7 μmol CO₂ m^?2 sec^?1), increased 38% under elevated CO₂, irrespective of combination with the drought or temperature treatments. In contrast, extended summer drought decreased R_S by 14%, while elevated soil temperature did not affect R_S overall. A significant interaction between elevated temperature and drought resulted in further reduction of R_S when these treatments were combined. A detailed analysis of short‐term R_S dynamics associated with drought periods showed that R_S was reduced by ~50% and was strongly correlated with soil moisture during these events. Recovery of R_S to pre‐drought levels occurred within 2 weeks of rewetting; however, unexpected drought effects were observed several months after summer drought treatment in 2 of the 3 years, possibly due to reduced plant growth or changes in soil water holding capacity. An empirical model that predicts R_S from soil temperature, soil moisture and plant biomass was developed and accounted for 55% of the observed variability in R_S. The model predicted annual sums of R_S in 2006 and 2007, in the control, were 672 and 719 g C m^?2 y^?1, respectively. For the full treatment combination, i.e. the future climate scenario, the model predicted that soil respiratory C losses would increase by ~21% (140–150 g C m^?2 y^?1). Therefore, in the future climate, stimulation of C storage in plant biomass and litter must be in excess of 21% for this ecosystem to not suffer a reduction in net ecosystem exchange.     

Effects of desiccation and rewetting on the release and decomposition of dissolved organic carbon from benthic macroalgae

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Insecticidal and feeding deterrent activities of essential oils in the cabbage looper,Trichoplusia ni (Lepidoptera: Noctuidae)

Ten essential oils were tested against the cabbage looper, Trichoplusia ni larvae for contact, residual and fumigant toxicities and feeding deterrent effects. Against third instar T. ni, Syzygium aromaticum (LD₅₀ = 47.8 μg/larva), Thymus vulgaris (LD₅₀ = 52.0 μg/larva) (the two positive controls) and Cinnamomum glanduliferum (LD₅₀ = 76.0 μg/larva) were the most toxic via topical application. Litsea pungens (LD₅₀ = 87.1 μg/larva), Ilex purpurea (LD₅₀ = 94.0 μg/larva), Cinnamomum cassia (LD₅₀ = 101.5 μg/larva) and Litsea cubeba (LD₅₀ = 112.4 μg/larva) oils were equitoxic. Thymus vulgaris (LC₅₀ = 4.8 mg/ml) and S. aromaticum (LC₅₀ = 6.0 mg/ml) oils were the most toxic in residual bioassays. Cymbopogon citratus (LC₅₀ = 7.7 mg/ml) and C. cassia (LC₅₀ = 8.5 mg/ml) oils were equitoxic followed by Cymbopogon nardus (LC₅₀ = 10.1 mg/ml) in this bioassay. The remaining five oils showed little or no residual effects. In a fumigation bioassay, L. cubeba (LC₅₀ = 16.5 μl/l) and I. purpurea (LC₅₀ = 22.2 μl/l) oils were the most toxic. Cinnamomum glanduliferum (LC₅₀ = 29.7 μl/l) and Sabina vulgaris (LC₅₀ = 31.2 μl/l) oils were equitoxic. Interestingly, S. aromaticum did not exhibit any fumigant toxicity. Cymbopogon citratus, C. nardus and C. cassia strongly deterred feeding by third instar T. ni (DC₅₀s = 26.9, 33.8 and 39.6 μg/cm², respectively) in a leaf disc choice bioassay. The different responses of T. ni larvae to the oils in different bioassays suggest that these essential oils exhibit different modes of action. Based on their comparable efficacy with essential oils already used as active ingredients in many commercial insecticides (i.e. clove oil and thyme oil), some of these essential oils may have potential as botanical insecticides against T. ni.     

Large‐scale screening and characterisation of Lemna aequinoctialis and Spirodela polyrhiza strains for starch production

• Duckweed is considered a promising feedstock for bioethanol production due to its high biomass and starch production. Selection of duckweed strains with high starch accumulation is essential for application of duckweeds to bioethanol production. Geographic differentiation had a large influence on genetic diversity of duckweeds.
• Biomass production, starch content and starch amount in geographically isolated strains of 20 Lemna aequinoctialis and Spirodela polyrhiza were calculated to evaluate their potential for bioethanol production. The influence of different collection time, culture medium and NaCl concentration on starch accumulation of the best strains were analysed.
• The results showed that biomass production, starch content and starch production of duckweeds demonstrated clonal dependency. The best strain was L. aequinoctialis 6000, with biomass production of 15.38 ± 1.47 g m^?2, starch content of 28.68 ± 1.10% and starch production of 4.39 ± 0.25 g m^?2. Furthermore, starch content of L. aequinoctialis 6000 was highest after 8 h of light, tap water was the best medium for starch induction, and NaCl did not induce starch accumulation.
• This study suggests duckweed biomass production and starch production demonstrate clonal dependency, indicating that extensive clonal comparisons will be required to identify the most suitable isolates for duckweed selective breeding for bioethanol.

     

Effects of human chorionic gonadotropin (hCG) and handling stress on spermiation of silver perch Leiopotherapon plumbeus (Kner, 1864)

This study determined the effect of human chorionic gonadotropin (hCG) and handling stress on the spermiation and milt response of silver perch Leiopotherapon plumbeus based on the measurement of spermatocrit, sperm density, and milt production. Compared to saline‐injected fish, the mean spermatocrit (or packed sperm) of hCG‐treated fish was significantly lower at 18 h (47.9%) and 30 h (40.2%) post‐injection while mean sperm density was significantly lower at 30 h post‐injection (3.6 × 10⁶ cells μl^?1) but not at 18 h. At 18 h (1.8 μl g‐BW^?1) and 30 h (2.5 μl g‐BW^?1) post‐injection, mean milt production of hCG‐treated fish was significantly higher than in the saline group. Milt consistency was also thinner in the hCG‐treated group. Mean sperm density of handled fish (18.0 × 10⁶ cells μl^?1) was significantly lower than control fish (23.4 × 10⁶ cells μl^?1). However, mean sperm density of handled plus saline‐injected (16.2 × 10⁶ cells μl^?1) and handled plus hCG‐treated fish (8.4 × 10⁶ cells μl^?1) was significantly lower than in the control goup. Having thicker milt consistency, mean spermatocrit and milt production of handled (77.5%; 1.1 μl g‐BW^?1, respectively) and handled plus saline‐injected fish (75.4%; 1.1 μl g‐BW^?1, respectively) were not significantly different from the control fish (76.2%; 1.3 μl g‐BW^?1, respectively). Handled plus hCG‐treated fish had the lowest mean sperm density (8.4 × 10⁶ cells μl^?1) and spermatocrit (54.7%), but had the highest mean milt production (5.5 μl g‐BW^?1) among the treatment groups. These results demonstrate that the hCG injection effectively induces spermiation and milt expression and that handling‐related stress negatively affects such responses. The spermatocrit method may be used to assess the spermiation and milt response of silver perch.     

Composition and Biological Activity of Picea pungens and Picea orientalis Seed and Cone Essential Oils

The increasing consumption of natural products lead us to discover and study new plant materials, such as conifer seeds and cones, which could be easily available from the forest industry as a waste material, for their potential uses. The chemical composition of the essential oils of Picea pungens and Picea orientalis was fully characterized by GC and GC/MS methods. Seed and cone oils of both tree species were composed mainly of monoterpene hydrocarbons, among which limonene, α‐ and β‐pinene were the major, but in different proportions in the examined conifer essential oils. The levorotary form of chiral monoterpene molecules was predominant over the dextrorotary form. The composition of oils from P. pungens seeds and cones was similar, while the hydrodistilled oils of P. orientalis seeds and cones differed from each other, mainly by a higher amount of oxygenated derivatives of monoterpenes and by other higher molar mass terpenes in seed oil. The essential oils showed mild antimicrobial action, however P. orientalis cone oil exhibited stronger antimicrobial properties against tested bacterial species than those of P. pungens. Effects of the tested cone essential oils on human skin fibroblasts and microvascular endothelial cells (HMEC‐1) were similar: in a concentration of 0 – 0.075 μl/ml the oils were rather safe for human skin fibroblasts and 0 – 0.005 μl/ml for HMEC‐1 cells. IC₅₀ value of Picea pungens oils was 0.115 μl/ml, while that of Picea orientalis was 0.105 μl/ml. The value of IC₅₀ of both oils were 0.035 μl/ml for HMEC‐1 cells. The strongest effect on cell viability had the oil from Picea orientalis cones, while on DNA synthesis the oil from Picea pungens cones.