Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its t...Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its toxicity are very few.This study investigated the toxicity of the O.cf.ovata(TIO991)isolated from Weizhou Island in the South China Sea by using methanol and chloroform to extract toxic compounds from the algal cells cultured indoor.Experiments on mouse acute toxicity showed that the crude methanol extract(CME)of O.cf.ovata caused the death of mice in 16–18 min.Furthermore,CME inhibited the cell reproduction of human neuroblastoma cells(BE(2)-M17 cells)by Cell Counting Kit-8 with a dose-and time-effect relationship and caused cell death in the form of cell necrosis.We found that CME had strong hemolytic activity and was significantly inhibited by ouabain,indicating that CME might contain palytoxins.By contrast,the crude chloroform extract of O.cf.ovata was relatively weak in toxicity as obtained in our experiments on mouse acute toxicity,cytotoxicity,and hemolytic activity.This suggests that the algae may raise the potential threat to marine ecosystems and public health.展开更多
[ Objective] The phytotoxicity effect of quinclorac on peanut succeeding seedlings was studied. [ Method] Peanut was taken as the indicator crop, plant height and fresh weight were adopted as bioassay indicator, the b...[ Objective] The phytotoxicity effect of quinclorac on peanut succeeding seedlings was studied. [ Method] Peanut was taken as the indicator crop, plant height and fresh weight were adopted as bioassay indicator, the biological activity of quinclorac on peanut was determined by the method of adding quinclorac in the soil, and the residue dynamic of quinclorac in paddy soil was determined. [Result] The linear correlation equations of peanut plant height and fresh weight with the concentration range of 0.7 -8.0 mg/kg separately were y = 11.235x ＋3.818 6, R^2 = 0.969 1 ; y = 5.973 3x ＋ 6.532 8, R^2 = 0.988 2. There would be no residual phytotoxicity effect when peanut was going to be planted in the same block in the second year. [Condusion] The bioassay method was simple and exact with good repeatability.展开更多
A simple in vivo bioassay suitable for the routine quality control testing of a new erythropoiesis stimulating protein was developed.Subcutaneous administration of the new erythropoiesis stimulating protein to Balb/c ...A simple in vivo bioassay suitable for the routine quality control testing of a new erythropoiesis stimulating protein was developed.Subcutaneous administration of the new erythropoiesis stimulating protein to Balb/c mice in a single dose resulted in a dose-dependent increase in the number of circulating reticulocytes.Within the erythropoiesis stimulating protein dose range of 3.125 to 200 ng per mouse,there is a strong linear relationship between the dose and reticulocyte counts in the treated mice.This linear relationship allows us to determine the biological potency of the testing erythropoiesis stimulating protein preparation relative to a reference standard using parallel line assay.Accuracy,precision,dose variation and blood collection time of this method were analyzed in order to choose doses in the linear range that are suitable for setting up a useful,precise,and economical bioassay.展开更多
Several natural and synthetic retinoids (vitamin-A derived analogies) were examined for their potential anti-cancer activity in both in vivo animal models and a novel in vitro human keratinocyte clonal growth bioassay...Several natural and synthetic retinoids (vitamin-A derived analogies) were examined for their potential anti-cancer activity in both in vivo animal models and a novel in vitro human keratinocyte clonal growth bioassay system. The natural retinoids included all-trans-retinoic (RA), 13-cis-retinoic acid, 4-oxoretinoic acid, and retinol. Among the synthetic retinoids tested were all trans N-(4-hydroxy(phenyl)retinamide, 3-substituted oxoretinoic acids, and 13 cis-N-ethylretinamide. The animal models employed were: 1) vitamin A-deficient hamster tracheal organ assay (HTOC);2) the benzo(α)pyrene-induced squamous metaplasia in a hamster tracheal organ system (BP-HTOC);3) the mouse skin tumor promoter (TPA)-induced ornithine decarboxylase enzyme assay(ODC);4) the mouse skin papilloma (MPA) assay;and 5) a novel retinoid bioassay in which retinoids display IC<sub>50</sub> values to inhibit clonal growth of NHK. All-trans-RA, 4-oxoretinoic acid and retinol were consistently more active than any of the synthetic derivatives in all bioassays tested. A statistical model was developed and significant positive correlations were found between: 1) ED<sub>50</sub> values in the HTOC system and reduction in TPA-induced ODC enzyme activity;2) tumors per animal in the MPA bioassay and suppression of TPA-induced ODC activity;and 3) a positive correlation between suppression of tumors per animal in the MPA assay, and retinoid inhibition of keratinocyte clonal growth. Test retinoids, were tested for their capacity to inhibit the clonal growth of a squamous carcinoma cell line (SCC-25), which were found to be 2 - 3 logs less sensitive for each tested retinoid than the corresponding activity against NHK cells. Antineoplastic retinoid drugs were reviewed.展开更多
Wastewater reclamation and reuse has been proved to be an effective way to relieve the fresh water crisis.However, toxic contaminants remaining in reclaimed water could lead to potential risk for reuse, and the conven...Wastewater reclamation and reuse has been proved to be an effective way to relieve the fresh water crisis.However, toxic contaminants remaining in reclaimed water could lead to potential risk for reuse, and the conventional water quality standards have difficulty guaranteeing the safety of reclaimed water.Bioassays can vividly reflect the integrated biological effects of multiple toxic substances in water as a whole, and could be a powerful tool for evaluating the safety of reclaimed water.Therefore, in this study, the advantages and disadvantages of using bioassays for evaluating the safety of reclaimed water were compared with those of conventional water quality standards.Although bioassays have been widely used to describe the toxic effects of reclaimed water and treatment efficiency of reclamation techniques, a single bioassay cannot reflect the complex toxicity of reclaimed water, and a battery of bioassays involving multiple biological effects or in vitro tests with specific toxicity mechanisms would be recommended.Furthermore, in order to evaluate the safety of reclaimed water based on bioassay results, various methods including potential toxicology, the toxicity unit classification system, and a potential eco-toxic effects probe are summarized as well.Especially, some integrated ranking methods based on a bioassay battery involving multiple toxicity effects are recommended as useful tools for evaluating the safety of reclaimed water, which will benefit the promotion and guarantee the rapid development of the reclamation and reuse of wastewater.展开更多
It has been difficult to perform dissolution test on solid preparations of traditional Chinese medicines （TCMs）. TCMs are different from chemical drugs in that their chemical compositions are complicated. The measur...It has been difficult to perform dissolution test on solid preparations of traditional Chinese medicines （TCMs）. TCMs are different from chemical drugs in that their chemical compositions are complicated. The measurement method based on chemical approach alone is incomplete. In order to solve this problem, in this study a bioassay-based dissolution test was developed. Microcalorimetry was used to obtain growth power-time curves and biothermodynamic parameters of Staphylococcus aureus inhibited by the solution of ShuangHuangLian （SHL） tablet, which was dissolved in phosphate buffer （pH 6.8） for different times. The results of the bioassay-based dissolution test of SHL tablet demonstrated that the bioassay method might be a promising alternative for its quality control.展开更多
An assessment method based on three toxicity tests （algae growth inhibition, daphnia immobilization and larval fish toxicity） was used to screen the biological safety of reclaimed water which was reused as sole repl...An assessment method based on three toxicity tests （algae growth inhibition, daphnia immobilization and larval fish toxicity） was used to screen the biological safety of reclaimed water which was reused as sole replenishment for scenic water system in a park （SOF Park） in northern China. A total of 24 water samples were collected from six sites of water system in the SOF Park in four different seasons. The results indicated that： （1） the reclaimed water directly discharged from a reclamation treatment plant near the SOF Park as influent of park had relatively low biological safety （all samples were ranked as C or D）; （2） the biological safety of reclaimed water was improved greatly with the ecological reclamation treatment processes composing of artificial wetland system and followed oxidation pond system; （3） the biological safety of reclaimed water in the main lake of SOF Park kept at a health status during different seasons （all samples were ranked as A）; （4） there was some certain correlation （R2= 0.5737） between the sum of toxicity scores and dissolved organic carbon for the studied water samples. It was concluded that the assessment method was reliable to screen the safety of reclaimed water reused as scenic water, and the reclaimed water with further ecological purification processes such as artificial wetland and oxidation pond system can be safely reused as scenic water in park.展开更多
Objective Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on industrial discharges, while the i...Objective Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on industrial discharges, while the impact of livestock wastes as a source of endocrine disrupters in aquatic environments has been rarely elucidated. In order to investigate the contribution of environmental estrogens from livestock, the estrogenic activity in water samples from a farm wastewater treatment plant was analyzed by a recombinant yeast screening method. Methods The extracts prepared from 15 selected water samples from the farm wastewater treatment plant, among which 6 samples were from pre-treatment section （influents） and 9 from post-treatment section （effluents）, were analyzed for estrogenic activity by cellar bioassay. Yeast cells transfected with the expression plasmid of human estrogen receptor and the Lac Z reporter plasmid encoding β-galactossidase, were used to measure the estrogen-like compounds in the farm wastewater treatment plant. Results The wastewater samples from influents showed a higher estrogenic potency than the effluent samples showing a low induction of β-galactossidase relative to solvent control condition. By comparison with a standard curve for 17β-estradiol （E2）, estrogenic potency in water samples from the influents was calculated as E2-equivalent and ranged from 0.1 to 150 pM E2-equivalent. The estrogenic potency in water samples from the effluents was significantly lower than that in the influents, and 7 water samples had less detectable limit in the total of 9 samples. Conclusion Yeast bioassay of estrogenic activity in most of the samples from the farm wastewater after disposal by traditional sewage treatment showed negative results.展开更多
Aim： To study the pharmacokinetic （PK） properties in rabbits treated with N-Ile^1-Thr^2-63-desulfato-r-hirudin （rH） newly developed in China by means of bioassay in order to provide preclinical experiment basis f...Aim： To study the pharmacokinetic （PK） properties in rabbits treated with N-Ile^1-Thr^2-63-desulfato-r-hirudin （rH） newly developed in China by means of bioassay in order to provide preclinical experiment basis for its development as a novel anticoagulant agent. Methods： rH plasma concentration was determined using bioassay based on ex vivo antithrombin activity of rH. Normal rabbits received iv rH 4.0, 2.0 and 1.0 mg/kg or sc rH 2.0 mg/kg, respectively. The rabbits with acute severe renal failure were given iv rH 2.0 mg/kg. Results： The bioassay described in this paper met requirements for study of PK in rabbits. The major PK parameters after iv dosing were as follows： t1/2p 58.4-59 min. Vd 0.09-0.12 L/kg, CL 0.0035-0.0040 L/（kg·min）; AUC were proportional to the doses, ti/2 and CL did not change significantly with the doses. The sc bioavailability reached 94%. The rabbits suffering from acute severe renal failure presented 11-fold longer t1/2p and 13-fold greater.4 UC than normal healthy rabbits. Conclusion： rH exhibited rapid elimination, distribution was only limited to extracellular space and good absorption from sc site. The excretion ofrH by kidneys played a very important role in the elimination of rH. The PK ofrH could be described by the two- and one-compartment model after iv and sc dosing, respectively, and folio,wed linear kinetics.展开更多
Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku;Phyllostachys bambusoides cv. Madake;P. nigra cv. Hachiku;Sasa kurilensis cv. Chishimazasa, which are of d...Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku;Phyllostachys bambusoides cv. Madake;P. nigra cv. Hachiku;Sasa kurilensis cv. Chishimazasa, which are of different classification or of different ecological distributions, using the “Sandwich Method”, which assays the dried leaves on growth of lettuce seedlings. Only small difference of activity was found among the four bamboo species. In addition, “Protoplast Co-culture Method” for assay of allelopathy in a 50 μL liquid medium using a 96 well culture plate, was applied to the suspension cultures of the four bamboo species. Protoplasts were isolated from two-week cultured suspension cells of four bamboo species using Cellulase RS and Pectolyase Y-23 in 0.6 M mannitol. At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory. Inhibitory effect of S. kurilensis was the strongest among four bamboo species. Furthermore, highly inhibitory effects of S. kurilensis protoplasts on yellow color accumulation of lettuce protoplasts were clearly observed by analysis of a scanned digital image of a 96-well culture plate. Differences and causes of the allelopathic activities were discussed comparing with other plant species studied using the same assay methods.展开更多
This study investigated the cytotoxicity of gemcitabine using the marine ciliate Euplotes vannus as the test organism.The median lethal concentrations(LC50 values)were determined using acute toxicity tests within an e...This study investigated the cytotoxicity of gemcitabine using the marine ciliate Euplotes vannus as the test organism.The median lethal concentrations(LC50 values)were determined using acute toxicity tests within an exposure time of 30 min with 0,6,12,24,and 48 mg mL^-1 gemcitabine.The median inhibition effect(IC50 value)on the growth of the ciliate cells was examined using chronic toxicity tests within 5 days(120 h)after exposure for 30 min with 0,0.7,3.5,7,and 14 mg mL^-1 gemcitabine.The 30-min LC50value was 10.66-mg mL 1.The LC50 values decreased with increasing exposure times and well fitted to the toxicity curve equation LC50=10.93+28.4e^-0.19t(R2=0.93;P<0.05,t=exposure time).The IC50 value for growth rates was 7.05 mg mL^-1,and the inhibition effect on growth rates well fitted to the model equation r%=0.8681e^-0.0782Cgem(r%means growth rate with inhibition by gemcitabine,Cgem means concentrations of gemcitabine,R^2=0.99 and P<0.05).The LC50 values of a wide range of gemcitabine concentrations could therefore be predicted for any given exposure time.These results suggest that the clinical dose of gemcitabine(20 mg mL^-1)was higher than the 30-min LC50 value,which was almost the same as the 6-min LC50 value(19.88 mg mL^-1)for E.vannus cells.The results also demonstrate that E.vannus can be used as a robust test organism for bioassays of chemotherapeutic drugs during short exposure periods.展开更多
Intensive agriculture activities,industrialization and growing numbers of wastewater treatment plants along river banks collectively contribute to the elevated levels of neurotoxic pollutants in natural water reservoi...Intensive agriculture activities,industrialization and growing numbers of wastewater treatment plants along river banks collectively contribute to the elevated levels of neurotoxic pollutants in natural water reservoirs across Europe.We established an in vitro bioassay based upon neural stem cells isolated from the subventricular zone of the postnatal mouse to evaluate the neurotoxic potential of raw wastewater,treated sewage effluent,groundwater and drinking water.The toxic potential of water samples was evaluated employing viability,proliferation,differentiation and migration assays.We found that raw wastewater could reduce the viability and proliferation of neural stem cells,and decreased the neuronal and astrocyte differentiation,neuronal neurite growth,astrocyte growth and cell migration.Treated sewage water also showed inhibitory effects on cell proliferation and migration.Our results indicated that relatively high concentrations of nitrogenous substances,pesticides,mercuric compounds,bisphenol-A,and phthalates,along with some other pollutants in raw wastewater and treated sewage water,might be the reason for the neuroinhibitory effects of these water samples.Our model successfully predicted the neurotoxicity of water samples collected from different sources and also revealed that the incomplete removal of contaminants from wastewater can be problematic for the developing nervous system.The presented data also provides strong evidence that more effective treatments should be used to minimize the contamination of water before release into major water bodies which may be considered as water reservoirs for human usage in the future.展开更多
Dried leaves of Prunus yedoensis and P. lannesiana (50 mg) showed strong inhibitory allelopathic activities, e.g., more than 97% growth inhibition of lettuce seedling using the sandwich method. Similarly, among suspen...Dried leaves of Prunus yedoensis and P. lannesiana (50 mg) showed strong inhibitory allelopathic activities, e.g., more than 97% growth inhibition of lettuce seedling using the sandwich method. Similarly, among suspension cultures induced from leaves and peduncles of two Prunus species, we found the strongest inhibitory allelopathic activities of protoplasts of leaf-origin suspension cells of P. yedoensis, when the protoplast co-culture method for bioassay of allelopathy was applied with lettuce as a recipient plant. Effects of two putative allelochemicals, abscisic acid and coumarin, on both protoplast cultures of lettuce and P. yedoensis were investigated. Coumarin inhibited the growth of lettuce protoplasts from low concentrations, while abscisic acid stimulated. Abscisic acid inhibited the protoplast growth of P. yedoensis from low concentrations, while coumarin did not, but inhibited only at a high concentration (1 mM). Contents of abscisic acid in protoplasts were measured using small scale purification and Enzyme Linked Immno Sorbent Assay, and contents of coumarin in leaf-origin susepension cells of P. yedoensis were measured using Gas Chromatography-Mass Spectrometry. Coumarin was more likely the allelochemical causing the strong inhibitory allelopathic activities of P. yedoensis in the protoplast co-culture bioassay. Effectiveness of the protoplast co-culture bioassay method of allelopathy was discussed.展开更多
The second-instar healthy larvae of Clostera. anastomosis were collected in the artificial woodland of poplar in Shuangcheng Town, Heilongjiang Province, China. The dead larvae of C. anastomosis infected by granulosis...The second-instar healthy larvae of Clostera. anastomosis were collected in the artificial woodland of poplar in Shuangcheng Town, Heilongjiang Province, China. The dead larvae of C. anastomosis infected by granulosis virus （GV） of Clostera anastomosis were grinded to obtain GV. The GV viral pesticide was diluted to seven concentrations, 1.58×10^3PIB·mL^-1, 1,58×10^4PIB·mL^-1, 1.58×10^5PIB·mL^-1 1.58×10^6PIB·mL^-1, 1,58×10^7PIB·mL^-1; 1.58×10^8PIB·mL^-1 and 1.58×10^9PIB·mL^-1 and the fresh poplar leaves were dipped in the seven concentrations liquids to feed the larvae. After nine days the mortality of larvae was investigated. The minimum corrected mortality （7.32%） of larvae was observed at concentrations of 1.58×10^3PIB·mL^-1 and the maximal mortality （97,36%） was observed at concentration of 1.58×10^9PIB·mL^-1. The regression equation between the logarithm of the virus concentration and the mortality was y= 1.946＋0.558x The LC50 was 2.97×10^5PIB·mL^-1. The LT50 for the virus concentration of 1.58×10^5, 1.58×10^6, 1.58×10^7, 1,58×10^58, 1.58×10^9 PIB·mL^-1 were 8.55d, 6.89d, 5.9d, 4.65d, and 4.08d, respectively, shorting gradually with the concentration increasing, It is concluded that the toxicity of Clostera anastomosis GV is very strong and as a kind of insecticides it has big potential in practical application.展开更多
An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing ...An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.展开更多
Allelopathy was defined in 1996 by IAS （international allelopathy society） as any process involving secondary metabolites produced by plant, algae, bacteria and fungi that influences the growth and development of ag...Allelopathy was defined in 1996 by IAS （international allelopathy society） as any process involving secondary metabolites produced by plant, algae, bacteria and fungi that influences the growth and development of agricultural and biological systems. Rice allelopathy against weeds was reported since 1989, which offers an integrated weed management with substantially reduced herbicide usage. Application of allelopathic rice cultivars is thought a resources conservation and environmental friendly way of weed bio-control, and could promote the sustainable development of agriculture. Screening or evaluating the allelopathic potential rice variety is the first step. In this paper, a new bioassay method was set up by the allelopathic potential of 9 rice lines on the target weed barnyardgrass （Echinochloa crusgalli）, and comparing with bioassay methods such as relay seeding in filter paper （RSF） and relay seeding in agar （RSA）. The results indicated that three methods had a same tendency in evaluating the allelopathic potential of rice; there existed a significant difference among different bioassay methods, and an interaction between bioassay methods and rice lines. The method of root exudates （RE） with the highest value and a correlation efficiency of 0.98 was considered as ideal bioassay method for evaluation of allelopathic potential.展开更多
The aim of this study was to develop and validate a simple,sensitive,precise and cost-effective onelevel agar diffusion（5＋1） bioassay for estimation of potency and bioactivity of Levofioxacin in pharmaceutical prep...The aim of this study was to develop and validate a simple,sensitive,precise and cost-effective onelevel agar diffusion（5＋1） bioassay for estimation of potency and bioactivity of Levofioxacin in pharmaceutical preparation which has not yet been reported in any pharmacopoeia.Among 16 microbial strains.Bacillus pumilus ATCC-14884 was selected as the most significant strain against Levofioxacin.Bioassay was optimized by investigating several factors such as buffer pH,inoculums concentration and reference standard concentration.Identification of Levofioxacin in commercial sample Levoflox tablet was done by FTIR spectroscopy.Mean potency recovery value for Levofioxacin in Levoflox tablet was estimated as 100.90%.A validated bioassay method showed linearity（r^2 = 0.988）,precision（Interday RSD=1.05%,between analyst RSD=1.02%） and accuracy（101.23%,RSD=0.72%）.Bioassay was correlated with HPLC using same sample and estimated potencies were 100.90% and 99.37%.respectively.Results show that bioassay is a suitable method for estimation of potency and bioactivity of Levofioxacin pharmaceutical preparations.展开更多
Objective To develop an ICR （female） mouse bioassay （MBA） for toxicity confirmation and evaluation of neurotoxins （brevetoxins）-contaminated shellfish. Methods Brevetoxins （BTX-B） as a causative agent of neuro...Objective To develop an ICR （female） mouse bioassay （MBA） for toxicity confirmation and evaluation of neurotoxins （brevetoxins）-contaminated shellfish. Methods Brevetoxins （BTX-B） as a causative agent of neurotoxic shellfish poisoning （NSP） under different shellfish matrices were intraperitoneally injected at different doses into mice to study their toxic effects and to differentiate the range of lethal and sublethal dosages. Their sensitivity and specificity were analyzed with 2 competitive ELISA kits for quantitative determination of standard BTX-B and dihydroBTX-B under different shellfish matrix-diluent combinations. Detection rates of MBA and two antibody-based assays for BTX-B from field NSP-positive shellfish samples were compared. Results BTX-B could be detected in shellfish tissues at concentration of 50-400 μg/100 g under shellfish matrix-Tween-saline media, which were appropriate to identify toxic shellfish at or above the regulatory limit （80 μg/100 g shellfish tissues）. The LD 50 identified was 455 g/kg for BTX-B under general shellfish matrices （excluding oyster matrices） dissolved in Tween-saline. The presence of shellfish matrices, of oyster matrices in particular, retarded the occurrence of death and toxicity presentation in mice. Two antibody-based assays, even in the presence of different shellfish matrix-diluent combinations, showed acceptable results in quantifying BTX-B and dihydroBTX-B well below the regulatory limit. Conclusion The two ELISA analyses agree favorably （correlation coefficient, r 0.96; Student＇s t-tests, P〉0.05） with the developed bioassay.展开更多
A method of extracting and purifying Cry1Ab protein(Bt toxin) from Cry1Ab transgenic rice was established. Most of the Bt toxin present in the tissue of Cry1Ab transgenic rice was extracted effectively with a solution...A method of extracting and purifying Cry1Ab protein(Bt toxin) from Cry1Ab transgenic rice was established. Most of the Bt toxin present in the tissue of Cry1Ab transgenic rice was extracted effectively with a solution of 50 mmol/LNa_2CO_3 and NaHCO_3. The crude protein containing Bt toxin was obtained after the pretreatment of Cry1Ab transgenic rice with ultra-filtration, ammonium sulfate precipitation and centrifugation. The dialysed crude protein was futher separated on DEAE Sephadex A-50 columns and Sephadex G-150 columns. The protein bound on DEAE Sephadex A-50 gel was eluted with buffer solution B(10 mmol/L tris-HCl buffer+1.0 mmol/L EDTA, pH=8.0) mixed with 0.1, 0.3, 0.5 and 0.8 mol/L NaCl in a discontinuous gradient elution mode. The peak of the Bt toxin eluted from the columns was identified by ELISA and bioassayed with larvae of tobacco hornworms and silkworms. The purity and the bioactivity of the Bt toxin were determined by means of SDS-PAGE and larvicidal assay, respectively. The purity of the Bt toxin obtained by this method is high, and its insecticidal activity is retained after the toxin is purified.展开更多
The joint action between chlorsulfuron and haloxyfop R was evaluated by bioassay with wheat and corn,respectivly.The dose response curve derived from wheat bioassay showed that the inhibition of haloxyfop R to whea...The joint action between chlorsulfuron and haloxyfop R was evaluated by bioassay with wheat and corn,respectivly.The dose response curve derived from wheat bioassay showed that the inhibition of haloxyfop R to wheat root growth wasn't affected by the increasing rate of chlorsulfuron.It indicated that chlorsulfuron had no antagonism to haloxyfop R.Meanwhile,the variation analysis of corn bioassay indicated that these two herbicides had joint action on inhibition to corn primary root growth.The joint action was evaluated as additive action by using Isobole Method.So chlorsulfuron and haloxyfop R could be used as tank mixture.展开更多
基金Supported by the Special Research for the Science and Technology Basic Resources Investigation Program of China(No.2018FY100200)the National Natural Science Foundation of China(No.42176206)the Natural Science Foundation of Shandong Province(No.ZR2021MD071)。
文摘Ostreopsis cf.ovata is a marine benthic dinoflagellate in tropical and temperate seas and can produce potent toxic compounds.The existence of O.cf.ovata has been found in the Chinese coastal areas,but studies on its toxicity are very few.This study investigated the toxicity of the O.cf.ovata(TIO991)isolated from Weizhou Island in the South China Sea by using methanol and chloroform to extract toxic compounds from the algal cells cultured indoor.Experiments on mouse acute toxicity showed that the crude methanol extract(CME)of O.cf.ovata caused the death of mice in 16–18 min.Furthermore,CME inhibited the cell reproduction of human neuroblastoma cells(BE(2)-M17 cells)by Cell Counting Kit-8 with a dose-and time-effect relationship and caused cell death in the form of cell necrosis.We found that CME had strong hemolytic activity and was significantly inhibited by ouabain,indicating that CME might contain palytoxins.By contrast,the crude chloroform extract of O.cf.ovata was relatively weak in toxicity as obtained in our experiments on mouse acute toxicity,cytotoxicity,and hemolytic activity.This suggests that the algae may raise the potential threat to marine ecosystems and public health.
文摘[ Objective] The phytotoxicity effect of quinclorac on peanut succeeding seedlings was studied. [ Method] Peanut was taken as the indicator crop, plant height and fresh weight were adopted as bioassay indicator, the biological activity of quinclorac on peanut was determined by the method of adding quinclorac in the soil, and the residue dynamic of quinclorac in paddy soil was determined. [Result] The linear correlation equations of peanut plant height and fresh weight with the concentration range of 0.7 -8.0 mg/kg separately were y = 11.235x ＋3.818 6, R^2 = 0.969 1 ; y = 5.973 3x ＋ 6.532 8, R^2 = 0.988 2. There would be no residual phytotoxicity effect when peanut was going to be planted in the same block in the second year. [Condusion] The bioassay method was simple and exact with good repeatability.
文摘A simple in vivo bioassay suitable for the routine quality control testing of a new erythropoiesis stimulating protein was developed.Subcutaneous administration of the new erythropoiesis stimulating protein to Balb/c mice in a single dose resulted in a dose-dependent increase in the number of circulating reticulocytes.Within the erythropoiesis stimulating protein dose range of 3.125 to 200 ng per mouse,there is a strong linear relationship between the dose and reticulocyte counts in the treated mice.This linear relationship allows us to determine the biological potency of the testing erythropoiesis stimulating protein preparation relative to a reference standard using parallel line assay.Accuracy,precision,dose variation and blood collection time of this method were analyzed in order to choose doses in the linear range that are suitable for setting up a useful,precise,and economical bioassay.
文摘Several natural and synthetic retinoids (vitamin-A derived analogies) were examined for their potential anti-cancer activity in both in vivo animal models and a novel in vitro human keratinocyte clonal growth bioassay system. The natural retinoids included all-trans-retinoic (RA), 13-cis-retinoic acid, 4-oxoretinoic acid, and retinol. Among the synthetic retinoids tested were all trans N-(4-hydroxy(phenyl)retinamide, 3-substituted oxoretinoic acids, and 13 cis-N-ethylretinamide. The animal models employed were: 1) vitamin A-deficient hamster tracheal organ assay (HTOC);2) the benzo(α)pyrene-induced squamous metaplasia in a hamster tracheal organ system (BP-HTOC);3) the mouse skin tumor promoter (TPA)-induced ornithine decarboxylase enzyme assay(ODC);4) the mouse skin papilloma (MPA) assay;and 5) a novel retinoid bioassay in which retinoids display IC<sub>50</sub> values to inhibit clonal growth of NHK. All-trans-RA, 4-oxoretinoic acid and retinol were consistently more active than any of the synthetic derivatives in all bioassays tested. A statistical model was developed and significant positive correlations were found between: 1) ED<sub>50</sub> values in the HTOC system and reduction in TPA-induced ODC enzyme activity;2) tumors per animal in the MPA bioassay and suppression of TPA-induced ODC activity;and 3) a positive correlation between suppression of tumors per animal in the MPA assay, and retinoid inhibition of keratinocyte clonal growth. Test retinoids, were tested for their capacity to inhibit the clonal growth of a squamous carcinoma cell line (SCC-25), which were found to be 2 - 3 logs less sensitive for each tested retinoid than the corresponding activity against NHK cells. Antineoplastic retinoid drugs were reviewed.
基金supported by the National Key R&D Program of China(Nos.2016YFE0118800,2018YFC0406301).
文摘Wastewater reclamation and reuse has been proved to be an effective way to relieve the fresh water crisis.However, toxic contaminants remaining in reclaimed water could lead to potential risk for reuse, and the conventional water quality standards have difficulty guaranteeing the safety of reclaimed water.Bioassays can vividly reflect the integrated biological effects of multiple toxic substances in water as a whole, and could be a powerful tool for evaluating the safety of reclaimed water.Therefore, in this study, the advantages and disadvantages of using bioassays for evaluating the safety of reclaimed water were compared with those of conventional water quality standards.Although bioassays have been widely used to describe the toxic effects of reclaimed water and treatment efficiency of reclamation techniques, a single bioassay cannot reflect the complex toxicity of reclaimed water, and a battery of bioassays involving multiple biological effects or in vitro tests with specific toxicity mechanisms would be recommended.Furthermore, in order to evaluate the safety of reclaimed water based on bioassay results, various methods including potential toxicology, the toxicity unit classification system, and a potential eco-toxic effects probe are summarized as well.Especially, some integrated ranking methods based on a bioassay battery involving multiple toxicity effects are recommended as useful tools for evaluating the safety of reclaimed water, which will benefit the promotion and guarantee the rapid development of the reclamation and reuse of wastewater.
基金National Natural Science Foundation of China (Grant No. 81073069)
文摘It has been difficult to perform dissolution test on solid preparations of traditional Chinese medicines （TCMs）. TCMs are different from chemical drugs in that their chemical compositions are complicated. The measurement method based on chemical approach alone is incomplete. In order to solve this problem, in this study a bioassay-based dissolution test was developed. Microcalorimetry was used to obtain growth power-time curves and biothermodynamic parameters of Staphylococcus aureus inhibited by the solution of ShuangHuangLian （SHL） tablet, which was dissolved in phosphate buffer （pH 6.8） for different times. The results of the bioassay-based dissolution test of SHL tablet demonstrated that the bioassay method might be a promising alternative for its quality control.
基金supported by the National High-Tech Research and Development Program (863) of China (No. 2008AA062502,2009AA063901)the Crucial Special Project:National Water Pollution Control and Management Science (No. 2008ZX07314-003: 008)the National Natural Science Foundation of China (No. 20877090,50938004)
文摘An assessment method based on three toxicity tests （algae growth inhibition, daphnia immobilization and larval fish toxicity） was used to screen the biological safety of reclaimed water which was reused as sole replenishment for scenic water system in a park （SOF Park） in northern China. A total of 24 water samples were collected from six sites of water system in the SOF Park in four different seasons. The results indicated that： （1） the reclaimed water directly discharged from a reclamation treatment plant near the SOF Park as influent of park had relatively low biological safety （all samples were ranked as C or D）; （2） the biological safety of reclaimed water was improved greatly with the ecological reclamation treatment processes composing of artificial wetland system and followed oxidation pond system; （3） the biological safety of reclaimed water in the main lake of SOF Park kept at a health status during different seasons （all samples were ranked as A）; （4） there was some certain correlation （R2= 0.5737） between the sum of toxicity scores and dissolved organic carbon for the studied water samples. It was concluded that the assessment method was reliable to screen the safety of reclaimed water reused as scenic water, and the reclaimed water with further ecological purification processes such as artificial wetland and oxidation pond system can be safely reused as scenic water in park.
基金the Natural Science foundation of Jiangsu Education Bureau (03KJB610168)
文摘Objective Environmental estrogens at an elevated concentration are known to produce adverse effects on human and animal life. However, the majority of researches have been focused on industrial discharges, while the impact of livestock wastes as a source of endocrine disrupters in aquatic environments has been rarely elucidated. In order to investigate the contribution of environmental estrogens from livestock, the estrogenic activity in water samples from a farm wastewater treatment plant was analyzed by a recombinant yeast screening method. Methods The extracts prepared from 15 selected water samples from the farm wastewater treatment plant, among which 6 samples were from pre-treatment section （influents） and 9 from post-treatment section （effluents）, were analyzed for estrogenic activity by cellar bioassay. Yeast cells transfected with the expression plasmid of human estrogen receptor and the Lac Z reporter plasmid encoding β-galactossidase, were used to measure the estrogen-like compounds in the farm wastewater treatment plant. Results The wastewater samples from influents showed a higher estrogenic potency than the effluent samples showing a low induction of β-galactossidase relative to solvent control condition. By comparison with a standard curve for 17β-estradiol （E2）, estrogenic potency in water samples from the influents was calculated as E2-equivalent and ranged from 0.1 to 150 pM E2-equivalent. The estrogenic potency in water samples from the effluents was significantly lower than that in the influents, and 7 water samples had less detectable limit in the total of 9 samples. Conclusion Yeast bioassay of estrogenic activity in most of the samples from the farm wastewater after disposal by traditional sewage treatment showed negative results.
基金Project (No. 96-C02-01-05A) supported by the National NaturalScience Foundation of China
文摘Aim： To study the pharmacokinetic （PK） properties in rabbits treated with N-Ile^1-Thr^2-63-desulfato-r-hirudin （rH） newly developed in China by means of bioassay in order to provide preclinical experiment basis for its development as a novel anticoagulant agent. Methods： rH plasma concentration was determined using bioassay based on ex vivo antithrombin activity of rH. Normal rabbits received iv rH 4.0, 2.0 and 1.0 mg/kg or sc rH 2.0 mg/kg, respectively. The rabbits with acute severe renal failure were given iv rH 2.0 mg/kg. Results： The bioassay described in this paper met requirements for study of PK in rabbits. The major PK parameters after iv dosing were as follows： t1/2p 58.4-59 min. Vd 0.09-0.12 L/kg, CL 0.0035-0.0040 L/（kg·min）; AUC were proportional to the doses, ti/2 and CL did not change significantly with the doses. The sc bioavailability reached 94%. The rabbits suffering from acute severe renal failure presented 11-fold longer t1/2p and 13-fold greater.4 UC than normal healthy rabbits. Conclusion： rH exhibited rapid elimination, distribution was only limited to extracellular space and good absorption from sc site. The excretion ofrH by kidneys played a very important role in the elimination of rH. The PK ofrH could be described by the two- and one-compartment model after iv and sc dosing, respectively, and folio,wed linear kinetics.
文摘Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku;Phyllostachys bambusoides cv. Madake;P. nigra cv. Hachiku;Sasa kurilensis cv. Chishimazasa, which are of different classification or of different ecological distributions, using the “Sandwich Method”, which assays the dried leaves on growth of lettuce seedlings. Only small difference of activity was found among the four bamboo species. In addition, “Protoplast Co-culture Method” for assay of allelopathy in a 50 μL liquid medium using a 96 well culture plate, was applied to the suspension cultures of the four bamboo species. Protoplasts were isolated from two-week cultured suspension cells of four bamboo species using Cellulase RS and Pectolyase Y-23 in 0.6 M mannitol. At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory. Inhibitory effect of S. kurilensis was the strongest among four bamboo species. Furthermore, highly inhibitory effects of S. kurilensis protoplasts on yellow color accumulation of lettuce protoplasts were clearly observed by analysis of a scanned digital image of a 96-well culture plate. Differences and causes of the allelopathic activities were discussed comparing with other plant species studied using the same assay methods.
基金supported by the National Natural Science Foundation of China (Nos. 31672308 and 40206021)
文摘This study investigated the cytotoxicity of gemcitabine using the marine ciliate Euplotes vannus as the test organism.The median lethal concentrations(LC50 values)were determined using acute toxicity tests within an exposure time of 30 min with 0,6,12,24,and 48 mg mL^-1 gemcitabine.The median inhibition effect(IC50 value)on the growth of the ciliate cells was examined using chronic toxicity tests within 5 days(120 h)after exposure for 30 min with 0,0.7,3.5,7,and 14 mg mL^-1 gemcitabine.The 30-min LC50value was 10.66-mg mL 1.The LC50 values decreased with increasing exposure times and well fitted to the toxicity curve equation LC50=10.93+28.4e^-0.19t(R2=0.93;P<0.05,t=exposure time).The IC50 value for growth rates was 7.05 mg mL^-1,and the inhibition effect on growth rates well fitted to the model equation r%=0.8681e^-0.0782Cgem(r%means growth rate with inhibition by gemcitabine,Cgem means concentrations of gemcitabine,R^2=0.99 and P<0.05).The LC50 values of a wide range of gemcitabine concentrations could therefore be predicted for any given exposure time.These results suggest that the clinical dose of gemcitabine(20 mg mL^-1)was higher than the 30-min LC50 value,which was almost the same as the 6-min LC50 value(19.88 mg mL^-1)for E.vannus cells.The results also demonstrate that E.vannus can be used as a robust test organism for bioassays of chemotherapeutic drugs during short exposure periods.
基金the Higher Education Commission(HEC),Pakistan(Ref.PD/OS-II/Batch-VI/Germany/2015/77159/17755)the University of Applied Sciences Kaiserslautern,Germany for providing financial support to execute the present project。
文摘Intensive agriculture activities,industrialization and growing numbers of wastewater treatment plants along river banks collectively contribute to the elevated levels of neurotoxic pollutants in natural water reservoirs across Europe.We established an in vitro bioassay based upon neural stem cells isolated from the subventricular zone of the postnatal mouse to evaluate the neurotoxic potential of raw wastewater,treated sewage effluent,groundwater and drinking water.The toxic potential of water samples was evaluated employing viability,proliferation,differentiation and migration assays.We found that raw wastewater could reduce the viability and proliferation of neural stem cells,and decreased the neuronal and astrocyte differentiation,neuronal neurite growth,astrocyte growth and cell migration.Treated sewage water also showed inhibitory effects on cell proliferation and migration.Our results indicated that relatively high concentrations of nitrogenous substances,pesticides,mercuric compounds,bisphenol-A,and phthalates,along with some other pollutants in raw wastewater and treated sewage water,might be the reason for the neuroinhibitory effects of these water samples.Our model successfully predicted the neurotoxicity of water samples collected from different sources and also revealed that the incomplete removal of contaminants from wastewater can be problematic for the developing nervous system.The presented data also provides strong evidence that more effective treatments should be used to minimize the contamination of water before release into major water bodies which may be considered as water reservoirs for human usage in the future.
文摘Dried leaves of Prunus yedoensis and P. lannesiana (50 mg) showed strong inhibitory allelopathic activities, e.g., more than 97% growth inhibition of lettuce seedling using the sandwich method. Similarly, among suspension cultures induced from leaves and peduncles of two Prunus species, we found the strongest inhibitory allelopathic activities of protoplasts of leaf-origin suspension cells of P. yedoensis, when the protoplast co-culture method for bioassay of allelopathy was applied with lettuce as a recipient plant. Effects of two putative allelochemicals, abscisic acid and coumarin, on both protoplast cultures of lettuce and P. yedoensis were investigated. Coumarin inhibited the growth of lettuce protoplasts from low concentrations, while abscisic acid stimulated. Abscisic acid inhibited the protoplast growth of P. yedoensis from low concentrations, while coumarin did not, but inhibited only at a high concentration (1 mM). Contents of abscisic acid in protoplasts were measured using small scale purification and Enzyme Linked Immno Sorbent Assay, and contents of coumarin in leaf-origin susepension cells of P. yedoensis were measured using Gas Chromatography-Mass Spectrometry. Coumarin was more likely the allelochemical causing the strong inhibitory allelopathic activities of P. yedoensis in the protoplast co-culture bioassay. Effectiveness of the protoplast co-culture bioassay method of allelopathy was discussed.
文摘The second-instar healthy larvae of Clostera. anastomosis were collected in the artificial woodland of poplar in Shuangcheng Town, Heilongjiang Province, China. The dead larvae of C. anastomosis infected by granulosis virus （GV） of Clostera anastomosis were grinded to obtain GV. The GV viral pesticide was diluted to seven concentrations, 1.58×10^3PIB·mL^-1, 1,58×10^4PIB·mL^-1, 1.58×10^5PIB·mL^-1 1.58×10^6PIB·mL^-1, 1,58×10^7PIB·mL^-1; 1.58×10^8PIB·mL^-1 and 1.58×10^9PIB·mL^-1 and the fresh poplar leaves were dipped in the seven concentrations liquids to feed the larvae. After nine days the mortality of larvae was investigated. The minimum corrected mortality （7.32%） of larvae was observed at concentrations of 1.58×10^3PIB·mL^-1 and the maximal mortality （97,36%） was observed at concentration of 1.58×10^9PIB·mL^-1. The regression equation between the logarithm of the virus concentration and the mortality was y= 1.946＋0.558x The LC50 was 2.97×10^5PIB·mL^-1. The LT50 for the virus concentration of 1.58×10^5, 1.58×10^6, 1.58×10^7, 1,58×10^58, 1.58×10^9 PIB·mL^-1 were 8.55d, 6.89d, 5.9d, 4.65d, and 4.08d, respectively, shorting gradually with the concentration increasing, It is concluded that the toxicity of Clostera anastomosis GV is very strong and as a kind of insecticides it has big potential in practical application.
基金Supported by a thesis research project of General Administration of Quality Supervision, Inspection and Quarantine of China (No. 2010IK168)
文摘An inter-laboratory comparison of the AOAC mouse bioassay for paralytic shellfish poisoning (PSP) toxicity in shellfish was carried out among 25 Chinese laboratories to examine the overall performance for PSP testing in China, and to analyze the main factors affecting the performance of this method. The toxic scallop Patinopecten yessoensis collected from coast of Bohai Sea, China, was used as a test sample in the comparison study. The results were reported and evaluated using robust statistical methods. The z scores showed that 80%, 8%, and 12% of laboratories reported satisfactory results, unsatisfactory results, and questionable results, respectively. This evaluation demonstrates that the PSP mouse bioassay is an appropriate method for screening and testing PSP toxicity in shellfish. However, it was found that the experience and skill of technicians, as well as the body weight and health status of mice being used significantly affected the accuracy of the method.
文摘Allelopathy was defined in 1996 by IAS （international allelopathy society） as any process involving secondary metabolites produced by plant, algae, bacteria and fungi that influences the growth and development of agricultural and biological systems. Rice allelopathy against weeds was reported since 1989, which offers an integrated weed management with substantially reduced herbicide usage. Application of allelopathic rice cultivars is thought a resources conservation and environmental friendly way of weed bio-control, and could promote the sustainable development of agriculture. Screening or evaluating the allelopathic potential rice variety is the first step. In this paper, a new bioassay method was set up by the allelopathic potential of 9 rice lines on the target weed barnyardgrass （Echinochloa crusgalli）, and comparing with bioassay methods such as relay seeding in filter paper （RSF） and relay seeding in agar （RSA）. The results indicated that three methods had a same tendency in evaluating the allelopathic potential of rice; there existed a significant difference among different bioassay methods, and an interaction between bioassay methods and rice lines. The method of root exudates （RE） with the highest value and a correlation efficiency of 0.98 was considered as ideal bioassay method for evaluation of allelopathic potential.
文摘The aim of this study was to develop and validate a simple,sensitive,precise and cost-effective onelevel agar diffusion（5＋1） bioassay for estimation of potency and bioactivity of Levofioxacin in pharmaceutical preparation which has not yet been reported in any pharmacopoeia.Among 16 microbial strains.Bacillus pumilus ATCC-14884 was selected as the most significant strain against Levofioxacin.Bioassay was optimized by investigating several factors such as buffer pH,inoculums concentration and reference standard concentration.Identification of Levofioxacin in commercial sample Levoflox tablet was done by FTIR spectroscopy.Mean potency recovery value for Levofioxacin in Levoflox tablet was estimated as 100.90%.A validated bioassay method showed linearity（r^2 = 0.988）,precision（Interday RSD=1.05%,between analyst RSD=1.02%） and accuracy（101.23%,RSD=0.72%）.Bioassay was correlated with HPLC using same sample and estimated potencies were 100.90% and 99.37%.respectively.Results show that bioassay is a suitable method for estimation of potency and bioactivity of Levofioxacin pharmaceutical preparations.
文摘Objective To develop an ICR （female） mouse bioassay （MBA） for toxicity confirmation and evaluation of neurotoxins （brevetoxins）-contaminated shellfish. Methods Brevetoxins （BTX-B） as a causative agent of neurotoxic shellfish poisoning （NSP） under different shellfish matrices were intraperitoneally injected at different doses into mice to study their toxic effects and to differentiate the range of lethal and sublethal dosages. Their sensitivity and specificity were analyzed with 2 competitive ELISA kits for quantitative determination of standard BTX-B and dihydroBTX-B under different shellfish matrix-diluent combinations. Detection rates of MBA and two antibody-based assays for BTX-B from field NSP-positive shellfish samples were compared. Results BTX-B could be detected in shellfish tissues at concentration of 50-400 μg/100 g under shellfish matrix-Tween-saline media, which were appropriate to identify toxic shellfish at or above the regulatory limit （80 μg/100 g shellfish tissues）. The LD 50 identified was 455 g/kg for BTX-B under general shellfish matrices （excluding oyster matrices） dissolved in Tween-saline. The presence of shellfish matrices, of oyster matrices in particular, retarded the occurrence of death and toxicity presentation in mice. Two antibody-based assays, even in the presence of different shellfish matrix-diluent combinations, showed acceptable results in quantifying BTX-B and dihydroBTX-B well below the regulatory limit. Conclusion The two ELISA analyses agree favorably （correlation coefficient, r 0.96; Student＇s t-tests, P〉0.05） with the developed bioassay.
基金Supported by the National Science Foundation of China(No.2 0 1770 2 1and 2 0 2 770 31)
文摘A method of extracting and purifying Cry1Ab protein(Bt toxin) from Cry1Ab transgenic rice was established. Most of the Bt toxin present in the tissue of Cry1Ab transgenic rice was extracted effectively with a solution of 50 mmol/LNa_2CO_3 and NaHCO_3. The crude protein containing Bt toxin was obtained after the pretreatment of Cry1Ab transgenic rice with ultra-filtration, ammonium sulfate precipitation and centrifugation. The dialysed crude protein was futher separated on DEAE Sephadex A-50 columns and Sephadex G-150 columns. The protein bound on DEAE Sephadex A-50 gel was eluted with buffer solution B(10 mmol/L tris-HCl buffer+1.0 mmol/L EDTA, pH=8.0) mixed with 0.1, 0.3, 0.5 and 0.8 mol/L NaCl in a discontinuous gradient elution mode. The peak of the Bt toxin eluted from the columns was identified by ELISA and bioassayed with larvae of tobacco hornworms and silkworms. The purity and the bioactivity of the Bt toxin were determined by means of SDS-PAGE and larvicidal assay, respectively. The purity of the Bt toxin obtained by this method is high, and its insecticidal activity is retained after the toxin is purified.
文摘The joint action between chlorsulfuron and haloxyfop R was evaluated by bioassay with wheat and corn,respectivly.The dose response curve derived from wheat bioassay showed that the inhibition of haloxyfop R to wheat root growth wasn't affected by the increasing rate of chlorsulfuron.It indicated that chlorsulfuron had no antagonism to haloxyfop R.Meanwhile,the variation analysis of corn bioassay indicated that these two herbicides had joint action on inhibition to corn primary root growth.The joint action was evaluated as additive action by using Isobole Method.So chlorsulfuron and haloxyfop R could be used as tank mixture.
