Detection of antigenic proteins expressed by lymphocystis virus as vaccine candidates in olive flounder, Paralichthys olivaceus (Temminck & Schlegel)

Although the major capsid proteins (MCPs) of lymphocystis disease virus (LCDV) have been characterized, little is known about the host-derived immune response to MCPs and other LCDV antigenic proteins. To identify antigenic proteins of LCDV that could be used as vaccine candidates in olive flounder, Paralichthys olivaceus, we analysed the viral proteins responsible for its virulence by applying immuno-proteomics. LCDV proteins were separated by one-dimensional gel electrophoresis, transferred to polyvinylidene difluoride membrane, and probed with homogeneous P. olivaceus antisera elicited by LCDV natural infection and vaccination with formalin-killed LCDV. Four immune-reactive proteins were obtained at 68-, 51-, 41- and 21 kDa using antisera collected from natural infection while two proteins at 51- and 21 kDa exhibited response to antisera from vaccinated fish, indicating that the latter two proteins have vaccine potential. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nanoelectrospray MS/MS, the 51 and 21 kDa proteins were identified as MCP and an unknown protein, respectively.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae

A study was conducted to evaluate the probiotic properties of endogenous rainbow trout microbiota against pathogenic Lactococcus garvieae. A total of 335 bacterial strains were isolated from rainbow trout and screened for antagonistic activity against L. garvieae using an agar spot assay. Antagonistic strains were grouped by PCR amplification of repetitive bacterial DNA elements (rep‐PCR) and identified by 16S rRNA gene sequence analysis. The results revealed that the antagonistic strains belonged to the genera Lactobacillus, Lactococcus and Leuconostoc. Further probiotic characteristics, such as specific growth rate, doubling time, resistance to biological barriers, antibiotic resistance, hydrophobicity and production of antimicrobial substances, were also studied. These strains were able to survive low pH and high bile concentrations, showed good adherence characteristics and a broad spectrum of antibiotic resistance. The antagonistic efficacy was maintained after sterile filtration and was sensitive to proteinase K, indicating that proteinaceous extracellular inhibitory compounds were at least partially responsible for pathogen antagonism. Based on these results, these strains should be further studied to explore their probiotic effects in challenge experiments in vivo. This study shows clear evidence that the indigenous trout‐associated microbiota may provide a defensive barrier against L. garvieae.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

基于神经网络预测的无线传感器网络田间射频信号路径损耗

为解决应用无线传感器网络技术监测农田信息时无法快速预测射频信号路径损耗的问题,基于神经网络理论研究了田间路径损耗与其影响因素间的关系。试验中选取915和2 470 MHz 2个载波频率,在冬小麦的不同生长阶段测量射频信号在田间各影响因素作用下的路径损耗,建立和验证基于神经网络的射频信号田间路径损耗预测模型。所建立模型模拟值与实测值的相关系数为0.92,应用建立的神经网络预测田间射频信号路径损耗并与实测值对比,最大预测误差绝对值为4.186 dB,最大预测标准差为2.759 dB,预测准确度为94.2%。所建立的BP网络可以对田间射频信号路径损耗进行预测。     

Toxicological effects of TiO2 and ZnO nanoparticles in soil on earthworm Eisenia fetida

Nanoparticles (NPs) of TiO₂ and ZnO are receiving increasing attention due to their widespread applications. To evaluate their toxicities to the earthworm Eisenia fetida (Savigny, 1826) in soil, artificial soil systems containing distilled water, 0.1, 0.5, 1.0 or 5.0 g kg⁻¹ of NPs were prepared and earthworms were exposed for 7 days. Contents of Zn and Ti in earthworm, activities of antioxidant enzymes, DNA damage to earthworm, activity of cellulase and damage to mitochondria of gut cells were investigated after acute toxicity test. The results from response of the antioxidant system combined with DNA damage endpoint (comet assay) indicated that TiO₂ and ZnO NPs could induce significant damage to earthworms when doses were greater than 1.0 g kg⁻¹. We found that Ti and Zn, especially Zn, were bioaccumulated, and that mitochondria were damaged at the highest dose in soil (5.0 g kg⁻¹). The activity of cellulase was significantly inhibited when organisms were exposed to 5.0 g kg⁻¹ of ZnO NPs. Our study demonstrates that both TiO₂ and ZnO NPs exert harmful effects to E. fetida when their levels are higher than 1.0 g kg⁻¹ in soil and that toxicity of ZnO NPs was higher than TiO₂.     

Fate of prions in soil: Degradation of recombinant prion in aqueous extracts from soil and casts of two earthworm species

Prions represent the active agent in transmissible spongiform encephalopathy (TSE) diseases and can remain infective to mammals even after prolonged periods in soil. The influence of mesofauna on prion dispersal and degradation in soil, however, remains unknown. In this study the effect of earthworms on the retention/dissemination of TSEs in soil was evaluated using a model recombinant prion protein (recPrP) and aqueous extracts from soil and fresh casts of two earthworm species, Lumbricus terrestris and Aporrectodea caliginosa. Our results showed that earthworm gut-derived enzymes did not enhance the degradation of recPrP in comparison to soil, even though non-prion related proteolytic activity was higher in fresh worm excrements than in soil samples. Complete degradation of recPrP occurred in the aqueous extracts from all samples within up to 6 days at +15 °C. The proteolytic enzymes responsible for degrading recPrP were inhibited by aprotinin and leupeptin and studies in pure cultures suggested these were most probably of soil microbial origin.     

Lethal Effects of Pulsed High-Voltage Discharge on Marine Plankton and Escherichia coli

Ballast seawater is considered globally as a major vector for invasions of non-indigenous organisms. Several technologies have been tested for their ability to remove organisms from ballast water. In the present study, we constructed a novel pulsed high-voltage discharge (PHVD) system that could operate in either high current mode with several hundred amperes or shockwave generating mode with relatively lower current in seawater. In laboratory-scale experiments, the PHVD system with shockwave-generating mode was found to be more effective in killing zooplankton (1.9- to 4.0-fold) and phytoplankton (3.3-fold) than high current mode at discharge with 300–500 pulses at 7.1 kV. Further experiments were carried out at different voltages and pulse-numbers to examine effects of the shockwave-generating PHVD system on viabilities of one zooplankton larva, two phytoplankton species, and an indicator bacterium suspended in seawater in a static chamber. For zooplankton, live cells were not detected at discharge with 400 pulses at 13 kV. For phytoplankton, the initial live cells of a dinoflagellate was decreased by 77?±?0.5%, and the initial chl a concentration of a diatom was decreased by 76?±?6% at discharge with 700 pulses at 13 kV. For an indicator bacterium Escherichia coli, live cells were not detected at discharges with 200 or 700 pulses at 13 kV. Measurements of ATP content of organisms showed congruent results with those obtained by the above methods, suggesting it may be a rapid method for evaluating treatment efficiency. Though further scale-up studies are necessary, these results suggest that the PHVD system have a high potential for applying to ballast seawater treatment.     

Viable seeds and spores of weed species in peat soil under pineapple cultivation*

Soil samples were taken from twenty-two fields in a peat area in W. Malaysia where pineapples had been grown for different periods. The numbers of viable seeds and spores in the top 15 cm of fields under cultivation for 6 months, 2, 6 and 10 years varied from 66–122 millions/ha. Of this total, 90% were spores of Nephrolepis biserrata and eight other ferns. The total numbers of seeds of the two sedges, one grass and nine dicotyledons recorded, ranged from 0–7 to 6.3 millions/ha. Les semences et spores viables de mauvaises herbes en sol tourbeux dans des plantations d'ananas Des échantillons de sols ont été prélevés dans vingt-deux champs dans une zone tourbeuse en Malaisie occidentale oü les ananas ont été cultivés pendant des périodes différentes. Les nombres de semences et spores viables dans les 15 premiers centimétres du sol dans les champs mis en culture depuis 6 mois, 2, 6 et 10 ans ont varié de 66 ä 122 millions/ha. Sur ce total, 90 %étaient des spores de Nephrolepis biserrata et de huit autres fougéres. Le nombre total de semences des deux roseaux, de la graminee et des neuf dicotylédones dont la présence dut notée, fut compris entre, 0,7 et 6,3 millions/ha. Lebensfähigen Unkrautsamen und sporen im Moorboden unter Ananas In einem Moor in W. Malaysia wurden aus 22 Feldern die unterschiedlich lang unter Ananas standen, Bodenproben entnommen. Die Zahl an lebensfähigen Samen und Sporen in den obersten 15 cm der Felder, die 6 Monate, 2, 6 und 10 Jahre lang unter Ananas standen, schwankte von 66 bis 122 Millionen je ha. Von dieser gesamten Menge waren 90% Sporen von Nephrolepis biserrata und acht anderen Farnen. Die Gesamtzahl an Samen von zwei Seggen, einem Gras und neun dikotylen Arten, belief sich auf 0,7 bis 6,3 Millionen jeha.     

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