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1.
本文对甜菜夜蛾核型多角体病毒的两个分离株(SeMNPV-M,SeMNPV-Z)的生物活性进行了比较,并对甜菜夜蛾核多角体病毒杀虫悬浮剂的田间应用效果进行了评估.病毒生物测定结果显示,SeNPV-M和SeNPV-Z感染三龄幼虫的半致死剂量(LD50)分别为195.8 PIBs/克饲料和242.4 PIBs/克饲料.使用6000PIB/克饲料的病毒剂量感染三龄幼虫,其半致死时间(LT50)分别为3.50d和3.68d.甜菜夜蛾病毒杀虫悬浮剂已在工厂生产,田问实验结果表明,甜菜夜蛾病毒杀虫悬浮剂可有效控制目标害虫的危害.  相似文献   

2.
为明确冬虫夏草线状菌体制剂的侵染力,进行了冬虫夏草线状菌体制剂对蝠蛾幼虫的感染试验。在相同条件下对蝠蛾幼虫进行人工接种,并对感染温度、菌剂用量和虫龄进行了优化。结果表明冬虫夏草线状菌体具有侵染蝠蛾幼虫的能力,优化后的感染条件为:菌体剂量为5 g(即5个标准剂量)、幼虫虫龄为3龄、培养温度为14℃。该结果为蝠蛾幼虫新型接种体的制备与开发提供了依据。  相似文献   

3.
本文对甜菜夜蛾核型多角体病毒的两个分离株(SeMNPV-M,SeMNPV-Z)的生物活性进行了比较,并对甜 菜夜蛾核多角体病毒杀虫悬浮剂的田间应用效果进行了评估。病毒生物测定结果显示,SeNPV-M和SeNPV-Z 感染三龄幼虫的半致死剂量(LD50)分别为195.8 PIBs/克饲料和242.4 PIBs/克饲料。使用6000PIB/克饲料的病 毒剂量感染三龄幼虫,其半致死时间(LT50)分别为3.50d和3.68d。甜菜夜蛾病毒杀虫悬浮剂已在工厂生产,田间 实验结果表明,甜菜夜蛾病毒杀虫悬浮剂可有效控制目标害虫的危害。  相似文献   

4.
小菜蛾Plutella xylostella是我国南方十字花科蔬菜上的重要害虫,已对田间常用的化学杀虫剂产生了严重的抗性。为寻找有效的小菜蛾生物防治措施,本实验研究了一株分离自家白蚁的玫烟色拟青霉Paecilomyces fumosoroseus (SCAU-PFCF01)对小菜蛾2~4龄幼虫的致病力。实验采用浸液法,供试浓度为1×103、1×104、1×105、1×106和1×107个孢子/mL。结果表明:随玫烟色拟青霉孢子浓度的升高,小菜蛾的感病死亡率增加,在浓度为1×107 /mL时,小菜蛾2、3和4龄幼虫的累计死亡率分别为96%、85%和80%。玫烟色拟青霉对小菜蛾各龄幼虫的致病力与供试龄期有关,其感病的敏感顺序为2龄、3龄和4龄。用时间 剂量 死亡率模型(time-dose-mortality model,TDM)对各龄幼虫的致病力数据进行模拟,所建模型均顺利通过Hosmer-Lemeshow拟合异质性检验,表明模型拟合良好,并由模型估计出了该菌株对小菜蛾各龄幼虫的致死剂量与致死时间。2龄幼虫接种后第7天、3龄幼虫接种后第5天、4龄幼虫接种后第4天的LC50估计值分别为1.17×104、1.44×104和5.21×104 /mL,LC90估计值分别为1.98×106、3.82×107和1.29×108 /mL。玫烟色拟青霉对小菜蛾幼虫的致死时间与浓度相关,供试各龄幼虫的LT50值随着孢子悬浮液浓度的增加而递减,在1×105~1×107 /mL的范围内,2龄幼虫的LT50值从3.16天降低到1.72天,3龄幼虫的LT50从3.21天降低到1.83天,4龄幼虫的LT50从3.69天降低到2.04天。即2龄幼虫致死所需的时间最短,其次为3龄幼虫,4龄幼虫致死所需的时间最长。结果显示了该株玫烟色拟青霉在小菜蛾的生物防治中具较强的应用潜力。  相似文献   

5.
从枣尺蠖(Sucra jujuba Chu)幼虫虫尸中分离到一种核型多核体病毒。其多角体平面图象多为四、五边形,直径约0.8—2.0微米,多数1.5微米。多角体在0.05MNa_2co_3+0.05MNacl 溶液中降解,能游离释放出杆状有毒粒子,病毒粒子两端圆滑,其大小约285—300×48—55毫微米。室内和野外在自然温度条件下试验结果表明,该株病毒对枣尺蠖幼虫有很高的致病力,室内感染3龄幼虫,其 LC_(50)为4×10~(4·6)多角体/毫升。但对槐尺蠖、木橑尺蠖、桑尺蠖、银纹夜蛾、家蚕和柞蚕不致病。  相似文献   

6.
结合室内生测、温室盆栽和大棚内小区试验的方法,探讨了寒区昆虫病原线虫(entomopathogenic nematode,EPN)对我国北方越冬韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang的防治作用。室内生测的结果显示,在侵染剂量为400条侵染期线虫(IJs)每头3龄幼虫或蛹时,异小杆线虫Heterorhabditis bacteriophora ZT(HbZT,哈尔滨市植物园分离株)、H.bacteriophora LG(Hb-LG,辽宁分离株)、H.bacteriophora NJ(Hb-NJ,美国新泽西分离株)和斯氏线虫Steinernema carpocapsae All(Sc-All)对迟眼蕈蚊3龄幼虫的致死率于72 h后均达100%,而对蛹的寄生效果则是Hb-ZT线虫最高;Hb-ZT与Sc-All(1∶1)混用,对幼虫的室内寄生效果高于单独施用2个线虫的寄生效果;温室盆栽试验中喷施Hb-ZT与Sc-All(1∶1)的线虫混合液,施用剂量减半(50万IJs/m2),对韭蛆成虫羽化的控制效果稍高于单独施用2个线虫(100万IJs/m2)效果;不同时期防治韭蛆的试验结果表明,于韭菜棚内发现韭蛆成虫510 d内喷施线虫的防治效果较好;韭菜大棚内喷施剂量为50万IJs/m2的Hb-ZT与Sc-All线虫混合液对韭蛆幼虫防效达100%,增产作用与常规化学药剂(辛硫磷)相当。  相似文献   

7.
Ali MEHRVAR 《昆虫学报》2013,56(6):708-714
在室内条件下调查了几种荧光增白剂对采自伊朗East Azarbaijan的棉铃虫Helicoverpa armigera核型多角体病毒两个地域株(EAZ-I 和EAZ-II)对棉铃虫2龄幼虫的杀虫活性, 以提高这两个地域株的生物学活性。结果表明:与EAZ-II相比, EAZ-I的杀虫活性强,其对棉铃虫幼虫的LC50和 LT50 值低(分别为1.98×103 OB/mL和122.7 h)。本研究所用的所有荧光增白剂均能有效增强病毒的生物学活性,特别是0.2% 的Tinopal F-3543与EAZ-I 混用对棉铃虫幼虫的LC50值最低(5.16×102 OB/mL),与病毒单独应用相比活性增强了3.84倍。幼虫致死的相对速率测定结果表明,荧光增白剂提高了菌株的LT50值,其中Tinopal F-3543 的效果最佳。这些结果说明,在害虫综合治理中影响围食膜通透性的荧光增白剂与核型多角体病毒制剂混用是一种可供选择的重要方法。  相似文献   

8.
将从山东省东营分离到的1株传染性法氏囊病病毒(IBDV)野毒(暂命名 IBDV SDDY株,经鉴定该株与 IB DV STC株具有较高的同源性)经 SPF 鸡胚传代,然后转为细胞培养,取第 20 代、21 代、25 代毒,以 1 倍剂量(3000TCID50/0.2mL )和5倍剂量不同的免疫剂量,分别在7日龄、14日龄对商品代海蓝褐蛋鸡进行免疫,并于免疫前用 IBD ELISA试剂盒检测 IBDV母源抗体水平,于 35 日龄用传染性法氏囊病病毒超强毒 (very virulent In fectious Bursal Disease Virus, vvIBDV )GX 8/99攻毒,在攻毒前再次检测IBDV的抗体水平,攻毒后观察记录各分组鸡的致病率和死亡率,并计算免疫器官体重指数,观察免疫器官的组织损伤情况。试验结果表明:3 代毒都具有较高免疫原性,但是20代毒仍具有较大的毒性,7 日龄接种会引起法氏囊的萎缩,造成持续的组织损伤;21 代毒、25代毒保护率高,无免疫抑制,是比较理想的疫苗来源;7日龄免疫较14日龄免疫更易造成组织损伤和免疫抑制,14日龄免疫较为合适。  相似文献   

9.
利用生物信息学软件对编码中蜂囊状幼虫病毒(Chinese sacbrood virus,CSBV)VP2结构蛋白的密码子进行优化来提高VP2蛋白表达量,并进行免疫原性研究确定VP2蛋白的免疫原性。通过对17株囊状幼虫病毒(SBV)和7株中蜂囊状幼虫病毒(CSBV)的VP2基因序列进行比对,选取代表毒株(GenBank:HM237361.1)CSBV-LN株VP2基因作为参考序列,并用在CSBV所有毒株中保守性相对较强的氨基酸替换CSBV-LN株VP2对应位置的氨基酸,通过在线软件(http://www.jcat.de/和http://genomes.urv.es/OPTIMIZER/)对其密码子进行优化,构建原核表达质粒pET-28a-optiVP2,并转化入BL21(DE3)中,进行诱导表达。表达产物经SDS-PAGE和Westren blot鉴定正确后,将表达蛋白纯化并免疫至白来杭母鸡,对免疫鸡的抗体水平和淋巴细胞增值指数进行检测,研究CSBV VP2蛋白的免疫原性。同时,收集免疫后鸡蛋制备卵黄抗体,经纯化再与CSBV相互作用后,接种于3日龄蜜蜂幼虫,观察接种后幼虫死亡率,研究卵黄抗体中和CSBV病毒能力。结果显示,优化后的optiVP2能够在大肠杆菌中高效表达,其表达产物具有良好免疫原性,表达产物接种白来杭母鸡可诱导产生抗体,制备的卵黄抗体具有中和CSBV的作用。本实验实现了CSBV结构基因VP2在原核系统的高效表达,鸡免疫实验表明其具有良好免疫原性,并且可诱导鸡产生保护作用中和抗体,为进一步开发防治CSBV的生物制剂提供了帮助。  相似文献   

10.
家蚕对马尾松毛虫质型多角体病毒的敏感性   总被引:2,自引:0,他引:2  
用虫体克隆技术,对马尾松毛虫质型多角体病毒湖南株(DpCPV-HN)进行了分离纯化,鉴定为质型多角体病毒1型。以家蚕春蕾×镇珠杂种F1代及自交的F2代4或5日龄幼虫进行毒力测定,以纯化的家蚕质型多角体病毒对F1代幼虫的毒力测定为对照。结果表明:家蚕品种春蕾×镇珠对家蚕质型多角体病毒敏感,马尾松毛虫质型多角体病毒湖南株能引起其感染发病;马尾松毛虫质型多角体病毒湖南株感染家蚕品种春蕾×镇珠F1代幼虫和F2代幼虫28天后的半致死剂量(LD50)分别为885个和18个CPB(质多角体),前者为后者的49倍。马尾松毛虫质型多角体病毒湖南株感染后的家蚕,其结茧率、化蛹率、羽化率、全茧量、茧层量和单蛾产卵数均有所下降,全茧量、茧层量、茧层率和单蛾产卵数与病毒感染剂量之间无显著关联。  相似文献   

11.
Virus yield produced by dead larvae of balsam fir sawfly, Neodiprion abietis (Harris) (Hymenoptera: Diprionidae), that had been infected at four different larval stages (second, third, fourth, or fifth instar) with two virus concentrations (10(5) polyhedral inclusion bodies (PIB) /ml or 10(7) PIB/ml), were analyzed and compared to determine the effects of instar and amount of virus inoculum on virus production. The results indicate that both larval stage and inoculation dosage significantly affect virus yield. On average, each dead larva produced 1.36-12.21 x 10(7) PIB, depending upon larval age and virus concentration of inoculation. Although each dead larva produced more PIB when it was inoculated in the fourth or fifth stage, inoculation of these larvae did not result in the highest virus yield because of low larval mortality. In terms of net virus return, third instars would maximize virus yield when they are inoculated with a virus concentration that can cause 95-100% larval mortality.  相似文献   

12.
目的寻找一种在最短时间内能为假丝酵母菌的鉴定提供最多信息的简易培养方法。方法在同一个平板内用分区划线法、多点接种法及小培养对标本进行培养,分离假丝酵母菌。结果一个平板内同时使用三种方法接种培养。可从分区划线法、多点接种法观察到菌落的全貌(如颜色、质地、形态)、种类、数量及生长特性等。可用显微镜从小培养中看到假丝酵母菌的孢子和菌丝的结构。结论由于假丝酵母菌种类繁多导致假丝酵母菌临床感染表现不同,临床诊断和治疗在很大程度上依赖于实验室的检查结果,因此寻找快速有效的鉴定方法至关重要。一个平板内同时用三种方法接种标本进行培养,可大大提高假丝酵母菌的检出率及鉴定效率,并能判定分离出的假丝酵母菌是目的菌株还是污染菌。  相似文献   

13.
Screening for cassava mosaic begomoviruses (CMBs)-resistance using grafting and whitefly inoculation was performed with local and improved cassava. The onset of symptom appearance and the evolution of Cassava mosaic disease (CMD) varied in function of genotypes and virus inoculation techniques used. Grafting position using cassava as scion or rootstock does not affect CMD display and evolution. No relation was established between the number of whiteflies feeding on each genotype and viral inoculation technique tested. Detopping of young leaves induces triggering effect on CMD expression. PCR and ELISA confirmed the EACMV-UG's preferential transmission by whitefly. Hypothesis of virus replication and cultivars's susceptibility were supported by virus increasing particles in infected cassava. Cultivars Mvuazi (TMSI 95/0528) and 96/1089A are suggested field immune to CMBs; Disanka (TMSI 95/0211), Yauma, Timolo, Bangi, Mahungu (TMS 92/297), Mvuama (TMS 83/138), Lueki (TMS 91/377) and Zizila (MV 99/0038) are CMD-resistant; whereas Ponjo, Lofiongi, Ngonga and Mboloko are susceptible. Our results showed that resistant genotypes may express CMD under high inoculum pressure such as grafting.  相似文献   

14.
The morphological, biochemical, and physiological characteristics of a phosphate solubilizing and antagonistic bacterial strain, designated as B0, isolated from a sub-alpine Himalayan forest site have been described. The isolate is gram negative, rod shaped, 0.8 x 1.6 microm in size, and psychrotrophic in nature that could grow from 0 to 35 degrees C (optimum temp. 25 degrees C). It exhibited tolerance to a wide pH range (3-12; optimum 8.0) and salt concentration up to 4% (w/v). Although it was sensitive to kanamycin, gentamicin, and streptomycin (<10 microg mL(-1)), it showed resistance to higher concentrations of ampicillin, penicillin, and carbenicillin (>1000 microg mL(-1)). The isolate showed maximum similarity with Pseudomonas putida based on 16S rRNA analysis. It solubilized tricalcium phosphate under in vitro conditions. The phosphate solubilization was estimated along a temperature range (4-28 degrees C), and maximum activity (247 microg mL(-1)) was recorded at 21 degrees C after 15 days of incubation. The phosphate solubilizing activity coincided with a concomitant decrease in pH of the medium. The isolate also exhibited antifungal activity against phytopathogenic fungi in Petri dish assays and produced chitinase, ss-l,3-glucanase, salicylic acid, siderophore, and hydrogen cyanide. The plant growth promotion and antifungal properties were demonstrated through a maize-based bioassay under greenhouse conditions. Although the bacterial inoculation was found to result in significant increment in plant biomass, it stimulated bacterial and suppressed fungal counts in the rhizosphere. The present study is important with respect to enumerating microbial diversity of the colder regions as well as understanding the potential biotechnological applications of native microbes.  相似文献   

15.
A phosphate solubilizing and antagonistic bacterial strain, isolated from a Western Ghat forest soil in Kerala province, India (designated as NII-0906), showed cold tolerance and grew from 10 to 37°C (optimum temperature 30°C). It was a Gram-positive, rod shaped, 0.8–1.6 μm in size, and exhibited tolerance to a wide pH range (5–12; optimum 7.0) and salt concentration up to 7% (w/v). The isolate showed maximum similarity with Exiguobacterium marinum TF-80T based on 16S rRNA analysis. It solubilized tricalcium phosphate under in vitro conditions. The phosphate solubilization was estimated along a temperature range (5–40°C), and maximum activity (84.7 μg mL−1 day−1) was recorded at 30°C after 10 days of incubation. The phosphate solubilizing activity coincided with a concomitant decrease in pH of the medium. The isolate also exhibited antifungal activity against phytopathogenic fungi in Petri dish assays and produced siderophore and hydrogen cyanide. The strain’s plant growth promotion properties were demonstrated through a cowpea-based bioassay under greenhouse conditions. The bacterial inoculation resulted in significant increment in plant root, stem and as well as in plant biomass. Further, scanning electron microscopic study revealed the root colonization in cowpea. These results could offer potential perspective for the strain to be used as plant growth-promoting rhizobacteria, which could be used as an inoculant for regional crops.  相似文献   

16.
The mean incubation period (time from inoculation with virus to first appearance of iridescence) was used as an indication of the rate of replication of Tipula iridescent virus (TIV) in Tipula oleracea larvae. The mean incubation period and survival time (time from inoculation with virus to death) were compared with the mean instar duration at a series of temperatures. In most stages of the insect the optimum temperature for the replication of TIV and the temperature for the shortest mean survival time coincided with the peak temperature (the temperature for the fastest development of the insect stage). The peak temperature for T. oleracea does not appear to be the same for each stage, and the optimum temperature for TIV replication appears to be closely linked to the peak temperature of the infected stage. The optimum temperature (the temperature at which most individuals survived from hatching to the adult stage) of the insect was 20°C. Tipula iridescent virus replicated in T. oleracea larvae and pupae at 3° and 27°C, which are near the temperature limits for the insect. Incubation periods and survival times in TIV-inoculated larvae incubated in the field were much longer in winter than in summer.  相似文献   

17.
A new strain of Friend recombinant mink cell focus-inducing retrovirus, FMCF -1-E, was found to induce leukemias in NFS and IRW mice. Although the isolate was obtained from a stock of FMCF -1 ( Troxler et al., J. Exp. Med. 148:639-653, 1978), FMCF -1-E was distinguishable from FMCF -1 by oligonucleotide fingerprinting and antigenic analysis, using monoclonal antibodies. These analyses suggested that FMCF -1-E is a distinct FMCF isolate rather than a simple variant of FMCF -1. After neonatal inoculation, the latency for leukemia induction was 3 to 8 months. A similar long latency was also seen when Friend murine leukemia virus 57 was inoculated into adult (6-week-old) IRW mice. However, sequential inoculation of FMCF -1-E at birth followed by Friend murine leukemia 57 at 6 weeks of age led to a shortened latency period (2.5 to 4 months). Only neonatal inoculation of Friend murine leukemia virus 57 was able to induce a more rapid appearance of leukemia. The leukemia cell type in the majority of cases, regardless of virus inoculation protocol, was erythroid, but occasional myeloid, lymphoid, and mixed leukemias were also observed. In contrast to NFS and IRW mice, BALB/c mice were resistant to leukemia induction by FMCF -1-E and also showed some transient resistance to leukemia induction by Friend murine leukemia virus 57.  相似文献   

18.
Ge J  Deng G  Wen Z  Tian G  Wang Y  Shi J  Wang X  Li Y  Hu S  Jiang Y  Yang C  Yu K  Bu Z  Chen H 《Journal of virology》2007,81(1):150-158
H5N1 highly pathogenic avian influenza virus (HPAIV) has continued to spread and poses a significant threat to both animal and human health. Current influenza vaccine strategies have limitations that prevent their effective use for widespread inoculation of animals in the field. Vaccine strains of Newcastle disease virus (NDV), however, have been used successfully to easily vaccinate large numbers of animals. In this study, we used reverse genetics to construct a NDV that expressed an H5 subtype avian influenza virus (AIV) hemagglutinin (HA). Both a wild-type and a mutated HA open reading frame (ORF) from the HPAIV wild bird isolate, A/Bar-headed goose/Qinghai/3/2005 (H5N1), were inserted into the intergenic region between the P and M genes of the LaSota NDV vaccine strain. The recombinant viruses stably expressing the wild-type and mutant HA genes were found to be innocuous after intracerebral inoculation of 1-day-old chickens. A single dose of the recombinant viruses in chickens induced both NDV- and AIV H5-specific antibodies and completely protected chickens from challenge with a lethal dose of both velogenic NDV and homologous and heterologous H5N1 HPAIV. In addition, BALB/c mice immunized with the recombinant NDV-based vaccine produced H5 AIV-specific antibodies and were completely protected from homologous and heterologous lethal virus challenge. Our results indicate that recombinant NDV is suitable as a bivalent live attenuated vaccine against both NDV and AIV infection in poultry. The recombinant NDV vaccine may also have potential use in high-risk human individuals to control the pandemic spread of lethal avian influenza.  相似文献   

19.
Potato virus X (PVX) isolated from the potato leaf and tuber samples which were collected from various fields in Damavand and Ardabil. The initial isolations of the virus were made from potato by mechanical inoculation on Gomphrena globosa L. and Chenopodium spp. that produce local lesion, and then it causes mosaic on Nicotiana spp. and Datura stramonium L. An isolate of the virus inoculated to Nicotiana glutinosa L. and it was maintained throughout the work. Sap from infected N. glutinosa was ineffective after dilution to 10-6, 10 minutes at 70 degrees and 10 weeks at room temperature. The virus was readily purified from infected leaves and the best protocol was Moreira & Jones 1980 than the other 2 methods of Fribourg 1975 and Shepard & Shalla 1972. Antisera were prepared against native, degraded proteins and micro precipitin test showed that both antisera had a 1/512 titer. Precipitin lines with D - Protein antiserum was better of the native protein antiserum in agar double diffusion test than treated with SDS. The isolate of the virus was not transmitted by none of 2 species of Cuscuta but transmitted from infected leaves to healthy plants with sap inoculation without using Carburandum. This isolate showed positive reaction with gamaglubulin in kate received from CIP centre.  相似文献   

20.
The rate of increase and doubling time of the HOB clone of Autographa californica nuclear polyhedrosis virus (AcMNPV-HOB) in neonate Trichoplusia ni larvae was determined by measuring the increase in viral DNA through time following inoculation with average doses of 50 or 17,400 occlusion bodies per larva. Changes in total DNA and viral DNA through time were followed by fluorescence spectroscopy and quantitative slot-blot DNA:DNA hybridization, respectively. Total DNA content (i.e., larval DNA and viral DNA) of larvae infected with the intermediate dose lagged behind that of noninfected larvae 30 hr post-inoculation (p.i), reached a maximum at 51 hr p.i., and stayed constant thereafter. The total DNA content of larvae inoculated with the high dose lagged behind that of the control group from 18 hr p.i. and increased slowly until death of the larvae (ca. 48 hr p.i.). The amount of viral DNA in larvae inoculated with the intermediate dose increased exponentially between 15 and 42 hr p.i., reached a maximum at 48 hr p.i., and stayed constant until 68 hr p.i., by which time most larvae had died. The amount of viral DNA in larvae inoculated with the high dose did not increase exponentially; initially the rate of increase was the same as that for larvae inoculated with the intermediate dose but became progressively lower after 13 hr p.i. Calculations of the rate of increase for AcMNPV-HOB in neonate T. ni larvae inoculated with the intermediate dose and incubated at 29 degrees C resulted in a value of 0.264 hr-1 (doubling time: 2.63 hr).(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

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