Insecticidal Activity and HPLC Correlation of Thuringiensin from Fermentation and Two-phase Aqueous Separation Processes

Thuringiensin produced by Bacillus thuringiensis subsp. darmstadiensis was further subjected to a two-phase aqueous separation system. A modified HPLC method and a test for quantitative pathogenicity using the house fly Musca domestica were used for analysis of thuringiensin. Within a realistic range of dosages, more effect was observed in the pupal stage than in the larval stage. The percentage effective control rate (ECR) was calculated by (100-percentage emergence); malformed and non-reproductive adults were considered as emerged. Pupal mortality, pupal weight, and ECR after feeding the three-day-old larvae were the measured response criteria for bioassay. The EC₅₀ of thuringiensin for pupae mortality was 1·64 μg ml^-1 diet, and 0·83 μg ml^-1 for mortality of adults. Insecticidal activity of the broth increased with fermentation time-course from 9th to 21st hour. The bioassay curve constructed with three-hour sampling interval during the fermentation course had good correlation to thuringiensin content as determined by the HPLC method. In the two-phase aqueous separation system, a maximum of 96·7% ECR was achieved with the bottom salt layer, compared to a value of 46·7% with the upper PEG layer. These results suggest that thuringiensin, prepared through a fermentation and recovery process, is suitable for pest control. © 1997 SCI.     

Assessment of potassium speciation in soil using traditional single leaching and modified sequential extraction processes

Purpose

The bioavailability of potassium (K) depends on its speciation distribution in the soil. Different methods are commonly used to estimate K speciation including traditional single leaching (TSL) and sequential extraction process (SEP). However, K speciation is largely affected by soil pretreatment methods. The effects of both TSL and SEP soil pretreatment methods were evaluated.

Materials and methods

The TSL method classifies K speciation content based on bioavailability, while the SEP classifies the metal speciation based on the effects of environmental conditions. These two methods, together with a modified sequential single leaching (SSL) scheme, were used to evaluate five types of soil including soil without potassium fertilization, soil with long-term K fertilization, alkaline soil, red soil, and forest soil. The soil samples were gathered randomly at depths varying up to 30 cm before being dried in air at room temperature. The samples were then ground and mixed before passing through a sieve (10 mesh or 100 mesh) in order to perform K speciation analysis via the modified SSL technique, the TSL method, and the four schemes of SEP.

Results and discussion

Soil pretreatment influenced K speciation, with higher concentration in soil samples sieved through 100 mesh than through 10 mesh. In alkaline soil, potassium was observed to be associated with carbonate. For the various SEP schemes, K speciation was found to be greatest in the residual fraction, with only 3% observed in the carbonate, exchangeable, metal organic complex, or amorphous hydroxides of Fe or Mn. After following the first two steps of the SEP schemes, the available K was similar to that of the TSL method. Distribution of non-exchangeable K using the TSL method was comparable with the five combined SEP extraction steps which were all affected by environmental conditions.

Conclusions

Pretreatment affected K speciation distribution and total amount of metal in the soil. The 100 mesh sieve was more effective in estimating K soil speciation. The SEP method was acceptable for estimating K speciation, with the Krishnamurti et al. (Analyst 120:659–665, 1995) scheme as a useful appraisal of K bioavailability. Combination analyses using both TSL and SEP methods are useful techniques to enable a better understanding of K speciation transformation in soil.

     

Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Antcin B and its ester derivative from Antrodia camphorata induce apoptosis in hepatocellular carcinoma cells involves enhancing oxidative stress coincident with activation of intrinsic and extrinsic apoptotic pathway

The triterpenoids methylantcinate B (MAB) and antcin B (AB), isolated from the medicinal mushroom Antrodia camphorata , have been identified as strong cytotoxic agents against various type of cancer cells; however, the mechanisms of MAB- and AB-induced cytotoxicity have not been adequately explored. This study investigated the roles of caspase cascades, reactive oxygen species (ROS), DNA damage, mitochondrial disruption, and Bax and Bcl-2 proteins in MAB- and AB-induced apoptosis of hepatocellular carcinoma (HCC) HepG2 cells. Here, we showed that MAB and AB induced apoptosis in HepG2 cells, as characterized by increased DNA fragmentation, cleavage of PARP, sub-G1 population, chromatin condensation, loss of mitochondrial membrane potential, and release of cytochrome c. Increasing the levels of caspase-2, -3, -8, and -9 activities was involved in MAB- and AB-induced apoptosis, and they could be attenuated by inhibitors of specific caspases, indicating that MAB and AB triggered the caspase-dependent apoptotic pathway. Additionally, the enhanced apoptotic effect correlates with high expression of Fas, Fas ligand, as well as Bax and decreased protein levels of Bcl-(XL) and Bcl-2, suggesting that both the extrinsic and intrinsic apoptosis pathways were involved in the apoptotic processes. Incubation of HepG2 cells with antioxidant enzymes superoxide dismutase and catalase and antioxidants N-acetylcysteine and ascorbic acid attenuated the ROS generation and apoptosis induced by MAB and AB, which indicate that ROS plays a pivotal role in cell death. NADPH oxidase activation was observed in MAB- and AB-stimulated HepG2 cells; however, inhibition of such activation by diphenylamine significantly blocked MAB- and AB-induced ROS production and increased cell viability. Taken together, our results provide the first evidence that triterpenoids MAB and AB induced a NADPH oxidase-provoked oxidative stress and extrinsic and intrinsic apoptosis as a critical mechanism of cause cell death in HCC cells.     

Eco‐efficiency as a strategy for optimizing the sustainability of pest management

Agricultural industrialization and the subsequent reliance on pesticides has resulted in numerous unintended consequences, such as impacts upon the environment and by extension human health. Eco‐efficiency is a strategy for sustainably increasing production, while simultaneously decreasing these externalities on ecological systems. Eco‐efficiency is defined as the ratio of production to environmental impacts. It has been widely adopted to improve chemical production, but we investigate the challenges of applying eco‐efficiency to pesticide use. Eco‐efficiency strategies include technological innovation, investment in research and development, improvement of business processes, and accounting for market forces. These components are often part of integrated pest management (IPM) systems that include alternatives to pesticides, but its implementation is often thwarted by commercial realities and technical challenges. We propose the creation and adoption of an eco‐efficiency index for pesticide use so that the broad benefits of eco‐efficient strategies such as IPM can be more readily quantified. We propose an index based upon the ratio of crop yield to a risk quotient (RQ) calculated from pesticide toxicity. Eco‐efficiency is an operational basis for optimizing pest management for sustainability. It naturally favors adoption of IPM and should be considered by regulators, researchers, and practitioners involved in pest management. © 2019 Society of Chemical Industry     

Plants as de-worming agents of livestock in the Nordic countries: historical perspective, popular beliefs and prospects for the future.

Preparations derived from plants were the original therapeutic interventions used by man to control diseases (including parasites), both within humans and livestock. Development of herbal products depended upon local botanical flora with the result that different remedies tended to develop in different parts of the world. Nevertheless, in some instances, the same or related plants were used over wide geographic regions, which also was the result of communication and/or the importation of plant material of high repute. Thus, the Nordic countries have an ancient, rich and diverse history of plant derived anthelmintic medications for human and animal use. Although some of the more commonly used herbal de-wormers were derived from imported plants, or their products, many are from endemic plants or those that thrive in the Scandinavian environment. With the advent of the modern chemotherapeutic era, and the discovery, development and marketing of a seemingly unlimited variety of highly efficacious, safe synthetic chemicals with very wide spectra of activities, herbal remedies virtually disappeared from the consciousness--at least in the Western world. This attitude is now rapidly changing. There is a widespread resurgence in natural product medication, driven by major threats posed by multi-resistant pest, or disease, organisms and the diminishing public perceptions that synthetic chemicals are the panacea to health and disease control. This review attempts to provide a comprehensive account of the depth of historical Nordic information available on herbal de-wormers, with emphasis on livestock and to provide some insights on potentially rewarding areas of "re-discovery" and scientific evaluation in this field.     

烟草生产全程机械化作业研究

为了探索烟叶生产过程中各机械的性能及工作效率,2013年在福建省建阳市莒口镇河坝新村开展了烟叶生产全程机械化作业试点研究。结果表明,起垄环节,碎土率方面邵武华友履带式起垄机表现最好(89.9%),沟底积土方面松溪永顺表现最好;覆膜环节,各测试机型表现中等,在效率方面优于人工覆膜;移栽环节,漏栽率上江苏久泰移栽机高于江苏井关移栽机;培土环节,漳州南冠培土机操作简便,掉头灵活,单人操作,单行双侧作业,深受烟农喜爱;拔杆环节,龙岩诚德拔杆机单人操作,操作简便,动力较足,拔杆顺畅,拔杆带土较少,效果较好,适应性较强。     

Specific characters of 16S rRNA gene and 16S–23S rRNA internal transcribed spacer sequences of Xylella fastidiosa pear leaf scorch strains

Pear leaf scorch, the only Xylella fastidiosa-induced disease reported from Taiwan, was found in area where the variety Hengshan (Pyrus pyrifolia) was grown. Strains of pear leaf scorch Xyl. fastidiosa (XF-PLS) shared similarities to strains of other host origins in the requirement of complex medium and the exhibition of rippled cell walls, however, recent serological and molecular biology studies showed difference among them. Five strains of XF-PLS were compared with 20 other strains originally isolated from almond, oleander, pecan, plum, peach, mulberry, grapes, citrus, coffee, and sycamore by sequence analyses of the 16S rRNA gene and 16S–23S rRNA internal transcribed spacer region (ITS). When sequences of 16S rRNA gene based on fragment size of 1,537–1,540 bp were compared, the similarity index among 5 XF-PLS strains was 99.3–99.8%, whereas it was 97.8–98.6% between XF-PLS strains and strains from other hosts. When sequences of 16S–23S rRNA ITS based on fragment size of 510–540 bp were compared, the similarity index among 5 XF-PLS strains was 99.0–100%, whereas it was 80.7–82% between XF-PLS strains and strains from other hosts. Multiple sequence alignments led to the identification of 5 polymorphic nucleotides in the 16S rRNA gene among the 25 Xyl. fastidiosa strains, and there were considerable variations in the nucleotide sequences of 16S–23S rRNA ITS between XF-PLS and the other 20 Xyl. fastidiosa strains. The phylogenetic trees revealed that XF-PLS strains were separated from strains of other hosts. Strains of other hosts were divided into four subgroups: strains from (1) oleander, (2) grape, almond M23 and mulberry, (3) citrus and coffee, and (4) pecan, peach, plum, sycamore and almond M12. Results indicate that XF-PLS strains were not closely related to the above-mentioned strains from other hosts and could possibly belong to a new subspecies of Xyl. fastidiosa.