成虫饲料中糖与奶粉不同配比对家蝇繁殖力及卵黄蛋白发生的影响

本文研究了成蝇饲料中糖与奶粉不同配比对家蝇繁殖力及卵黄蛋白发生的影响,旨在为家蝇Musca domestica大规模高效养殖提供技术支持。结果表明,随着成蝇饲料中奶粉比例提高,单雌产卵量逐渐提高,且差异显著,但当饲料中奶粉比例达到和超过80%以后,单雌产卵量又显著减少,其中以60%奶粉+40%白糖饲养雌蝇产卵量最高(1935.83粒/雌),用100%白糖饲养产卵量最低(328.17粒/雌)。当成蝇饲料中白糖的添加比例为20%~60%时,雌虫寿命和产卵期差异均不显著,但用100%奶粉或80%以上的白糖饲养,雌虫寿命和产卵期显著缩短;雄虫寿命则随饲料中白糖比例的增加而延长;奶粉与白糖配比的变化对雌蝇产卵前期无显著影响。卵孵化率随饲料中白糖比例的增加而下降,尤其当饲料中白糖比例超过60%以后,卵孵化率下降更为明显。进一步测定卵黄蛋白的结果表明,雌蝇取食含糖量越高的饲料,其每日合成的卵黄蛋白及羽化后15 d内合成的总卵黄蛋白也越少,尤其是雌蝇完全摄取白糖,其每日合成的卵黄蛋白均维持在一个较低的水平,而蛋白质补充过多(如奶粉在饲料中的比例超过60%),雌蝇每日合成的卵黄蛋白及羽化后15 d内合成的总卵黄蛋白,基本不随饲料中奶粉的增加而增加。综合以上结果,作者认为用60%奶粉+40%白糖作为家蝇成虫的营养补充料,其不仅能保持雌蝇良好的繁殖力,同时也能有效降低成蝇养殖成本。     

温度对蜘蛛繁殖力和实验种群的影响

从我国常见10科18种蜘蛛的温度实验数据中,找出温度对蜘蛛繁殖和实验种群增长影响的一般规律和不同种类蜘蛛繁殖的最佳温度。根据实验可以看出：大多数蜂蛛在1－35℃温区内均可产卵和孵化;由于蜘蛛种类不同,雌蛛产卵袋数最多、卵袋内含卵量最高、产卵间隔时间最短、产卵率最高、单雌产卵量最多、孵化率最高和种群增长指数最大的最适温度也表现出差异;大多数种类雌蛛产卵袋数最多的温区是25－32℃;产卵间隔最短的温区是30－37℃;产卵率最高的温区是25－32℃;孵化率最高的温区在25℃左右;种群数量增长最多的温区是25－29℃。温度与蜘蛛繁殖力的关系可以用曲线方程和数学模型表示。     

家蝇卵巢在体外培育中摄取卵黄蛋白

本文报道家蝇Musca domestica卵巢在体外培养条件下,摄取异硫氰萤光素标记的家蝇卵黄蛋白的特点。用Grace's培养液标记蛋白浓度为2mg/m1,在27℃条件下培养2小时,卵巢摄取量依赖于培养液中卵黄蛋白浓度和温度,摄取高峰在羽化后48小时,正值卵母细胞发育阶段进入6-8时期。培养液中加入JHIII,能促进摄取,JHIIl的浓度和摄取量无明显相关性。乌本苷、牛血清蛋白和叠氮钠显著抑制卵巢的摄取活动。     

家蝇卵黄蛋白基因启动子区的克隆与活性分析

从家蝇基因组文库中分离到含约1．7kb5’上游区的家蝇卵黄蛋白-1基因组基因序列,根据其5’上游序列,PCR扩增出大小不同的4个启动子片段,分别插入到切除了CMV启动子的pCMV-GFP质粒中的绿色荧光蛋白报告基因上游,构建了pMYP1-GFP、pMYP2-GFP、pMYP3-GFP和pMYP4-GFP4个重组质粒。另将 684／ 7、 1165／ 7这两个启动子片段用SpeⅠ和HindⅢ双酶切,去除包含CAAT／TATA盒的 302／ 7序列区后,分别构建了pMYP5-GFP和pMYP6-GFP两个重组质粒。通过电转移实验和荧光检测表明。 684／ 7、 1165／ 7、 1616／ 7这3个启动子片段具有转录活性,而 684／ 7启动子片段的转录活性最强, 296／ 7、 684／ 302、 1165／ 302这3个启动子片段无转录活性。上述实验结果表明。 302／ 7序列区为启动子的核心部分。 302到 1616之间存在调控性启动子或增强子等其他一些顺式元件。细胞转染实验证实,6种启动子片段在BHK-21和Sf9细胞中都未表现出可检测的转录活性,说明家蝇卵黄蛋白基因启动子具有组织或细胞特异性。     

家蝇卵黄蛋白基因的克隆与结构序列分析

家蝇卵黄蛋白基因编码的卵黄蛋白是家蝇胚胎发育的重要营养来源 .根据 3种家蝇卵黄蛋白cDNA保守序列设计引物 ,用PCR技术从家蝇基因组DNA中扩增到大小为 76 8bp的mdYP1基因的部分DNA片段 .经地高辛标记成特异性探针 ,从构建的家蝇基因组文库中筛选出一个阳性克隆 ,并从该克隆中分离到大小为 3991bp的mdYP1基因组基因 .序列分析显示 ,该基因组序列含有约1 6kb的 5′ 上游区和 1 0kb的 3′ 下游区 ,编码区由一个 6 1bp的内含子和大小分别为 2 2 2bp和10 2 8bp的 2个外显子组成 .5′ 上游区含有典型的CAAT TATA盒 .     

黄粉虫成虫繁殖力的研究

黄粉虫因其易于饲养和具有高营养价值,而成为动物蛋白饲料。提供良好的饲养条件,用科学方法充分发挥成虫的繁殖力,使雌虫多产高质量的卵,是黄粉虫饲养过程中的重要环节,因此也是本研究所要探讨的内容。     

温度对黄粉虫成虫繁殖的影响

黄粉虫是多种小型经济动物的常用优良饵料,本试验在20.1℃,24.0℃、28.5℃、31.7℃和36.5五种恒温下饲养该成虫结果,成虫寿命平均分别为63.0、54.2、38.8、38.0和26.1天;每雌平均产卵量则分别为200.3、207.3、122.3、115.8和81.2粒,成虫平均生产1g卵消耗麸皮量分别为2.66、2.00、2.19、2.14和4.15g。结果说明人工繁殖黄粉虫的成虫期温度以24℃为最适宜。     

家蝇卵黄蛋白基因的克隆与结构序列分析

解放军军需大学军事兽医研究所夏平安、江禹和中国农业大学生物学院刘维全等研究人员对此课题作了研究,他们认为家蝇卵黄蛋白基因编码的卵黄蛋白是家蝇胚胎发育的重要营养来源。根据3种家蝇卵黄蛋白cDNA保守序列设计引物,用PCR技术从家蝇基因组DNA中扩增到大小为768bp的mdyp1基因的部分DNA片段,     

家蝇性别,阳光及温度对灭蝇灯杀灭效果的影响

灭蝇灯是1种以黑光灯诱蝇并用高压电网击杀的灭蝇器械,为了提高灭蝇灯的杀灭效果及测试方法,我们进行这项实验。1材料与方法1．1灭蝇灯：实验用3种灭蝇灯均用220V市电,功率40W。灭蝇灯内设高压电网,其电压均大于1000V（1）MFCQ9100型灭蝇灯,53cm×13cm×36cm,由武汉市电视机配件厂生产;（2）豪华型依丽安灭蝇灯,50cm×10cm×37cm,珠海依丽安电子厂提供;（3）DY、M-Ⅱ型灭蝇灯,70cm×17Cm×46cm,由南京星晨公司提供。1．2试虫：家蝇MuscadomesticaL．由本单位饲养室提供。在研究性别影响的实验中用乙醚低温麻醉法分雌雄,…     

Evaluation of Beauveria bassiana applications against adult house fly, Musca domestica, in commercial caged-layer poultry facilities in New York state

Applications of a commercially produced Beauveria bassiana product, balEnce, were compared with pyrethrin treatments for the control of adult house flies in New York high-rise, caged-layer poultry facilities. An integrated fly management program, which included the release of house fly pupal hymenopteran parasitoids, was used at all facilities. Adult house fly populations were lower in B. bassiana-treated facilities during the spray and post-spray periods, as recorded on spot cards. Concurrently, the numbers of house fly larvae recovered in B. bassiana-treated facilities were less than one-half that of the pyrethrin-treated facilities. House fly pupal parasitism levels were low, but similar under both treatment regimes. The numbers of adult and larval Carcinops pumilio, a predatory beetle, recovered from B. bassiana-treated facilities were 43 and 66% greater than from the pyrethrin-treated facilities, respectively.     

Host age, sex, and pathogen exposure level as factors in the susceptibility of Musca domestica to Entomophthora muscae

Adult Musca domestica were exposed to conidial showers from cadavers that were killed by Entomophthora muscae. Of the 4 host ages (14 days, 7 days, 3 days and <1 day), older flies were more likely to die without postmortem signs of infection, while younger flies were more likely to die with signs and produce primary conidia. The youngest fly group also succumbed to infection significantly sooner than older flies. Total mortality did not differ for the 2 sexes, but females died significantly more often with signs in one trial. Males died significantly earlier than did females, but differences were not significant when the sexes were paired by weight. Increased levels of pathogen exposure significantly increased both total mortality and the number of flies dying with signs; it also decreased incubation period. The potential significance of these findings in the epidemiology of the disease is discussed.

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Zusammenfassung Die Mortalitätsrate von Musca domestica nach Behandlung mit Entomophthora muscae Sporen wurde in Laborversuchen in Abhängigkeit von Wirtsalter,-geschlecht und Expositionsdauer festgestellt. Fliegen verschiedenen Alters (14 Tage, 7 Tage, 3 Tage und jünger als 1 Tag) wurden gemeinsam in 237 ml Kartonbehältern gehalten. Die Behälter besassen Netzdeckel, auf denen frisch an einer Entomophthora Infektion gestorbene Fliegen ausgelegt wurden. Auf diese Weise wurden die Versuchstiere 8 Stunden (low exposure) bzw. 48 Stunden (high exposure) lang mit Entomophthora Sporen kontaminiert. Mit Hilfe von am Boden der Versuchsgefässe ausgelegten Objektträgern wurde die relative Menge Entomophthora Sporen festgestellt, der die Versuchstiere ausgesetzt waren. Der Totenfall wurde täglich für die Dauer von 10 Tagen registriert; dabei wurde zwischen Konidienträgern (external signs) und Leichen ohne Konidien (no external signs) unterschieden.Die Versuchsgruppen, die 48 Stunden lang mit Entomophthora Sporen behandelt worden waren, wiesen im Vergleich zu den kürzer behandelten Versuchsgruppen höhere Mortalität und einen höheren Anteil an Konidienträgern auf. Darüber hinaus wurde das Auftreten von Konidienträgern bei den länger behandelten Gruppen früher beobachtet. Im Vergleich zu unbehandelten Kontrollgruppen erhöhte die Behandlung mit Entomophthora Sporen auch die Rate nicht konidientragender Leichen.Bei gleicher Expositionsdauer zeigten Weibchen und Männchen ähnliche Mortalitätsraten. Ein höherer Anteil Konidienträger bei den Weibchen wurde bei einem von zwei entsprechenden Versuchen beobachtet. Konidienträger traten bei Weibchen später als bei Männchen auf. In Bezug auf die Inkubationszeit konnte kein geschlechsspezifischer Unterschied festgestellt werden, wenn Versuchsgruppen mit gleichem Gewicht verglichen wurden. Bei beiden Geschlechtern erfolgte die Bildung von Konidien bei grösseren Individuen später als bei kleineren Individuen.Das Alter der Versuchstiere beeinflusste sowohl die Mortalitätsrate als auch die Häufigkeit und das zeitliche Auftreten von Konidienträgern. Jüngere Fliegen wiesen einen höheren Anteil Konidienträger auf; bei Fliegen mit einem Alter von weniger als einem Tag wurde das Auftreten von Konidienträgern früher beobachtet als bei älteren Individuen.Die untersuchten Faktoren können auch für die Epidemiologie von Entomophthora freilebender Musca domestica Populationen eine wichtige Rolle spielen. In Populationen mit niedrigerem Durchschnittsalter wird sich eine Infektion rascher ausbreiten, da jüngere Fliegen eine kürzere Inkubationszeit aufweisen und auch nach kürzerer Kontaminationsdauer Konidien an den getöteten Individuen ausgebildet werden. Darüber hinaus wird sich eine Infektion dann rascher ausbreiten, wenn die Population mit einer grösseren Sporenmenge infiziert wird, da dann eine kürzere Inkubationzeit und ein höherer Anteil Konidienträger zu erwarten ist.

     

Map position of Aldox (Aldehyde-oxidase) and Adh (Alcohol dehydrogenase) loci on autosome II of Musca domestica L.

The Aldox and Adh structural loci of Musca domestica L. belong to autosome II. They code for the enzymes aldehyde oxidase and alcohol dehydrogenase. Both these enzymes have allelic variants with specific electrophoretic mobility, which, on cellogel, are seen as single bands. The Aldox and Adh loci encompass a large map interval, which includes the morphological markers ar, cm, and car. The recombination frequencies between these five loci indicate the alignment Aldox-ar-cm-car-Adh.

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Localisation chez Musca domestica L. des gènes Aldox et Adh codant les enzymes ald\;ehyde oxydase (AO) et alcool déshydrogénase (ADH)

Résumé Chez la mouche, les enzymes AO et ADH, observées par électrophorèse sur cellogel, présentent toutes 2 des formes alléliques exprimées par des bandes ayant une mobilité anodique propre.Les gènes structuraux Aldox et Adh, codant ces formes, sont liés entre eux et situés sur le chromosome 2. Ils se recombinent avec une fréquence élevée d'interchange; ils sont donc séparés par un intervalle important dans lequel sont compris les caractères visibles ar, cm, car. La fréquence des recombinaisons entre caractères visibles et gènes enzymatiques indique l'ordre suivant sur ce segment du deuxième chromosome de la mouche: Aldox, ar, cm, car, Adh.

     

Knockdown resistance (kdr) to DDT and pyrethroid insecticides maps to a sodium channel gene locus in the housefly (Musca domestica)

The voltage-sensitive sodium channel is generally regarded as the primary target site of dichlorodiphenyl-trichloro-ethane (DDT) and pyrethroid insecticides, and has been implicated in the widely reported mechanism of nerve insensitivity to these compounds. This phenomenon is expressed as knockdown resistance (kdr) and has been best characterised in the housefly where several putative alleles, including the more potent super-kdr factor, have been identified. We report the isolation of cDNA clones containing part of a housefly sodium channel gene, designated Msc, which show close homology to the para sodium channel of Drosophila (99% amino acid identity within the region of overlap). Using Southern blots of insect DNA, restriction fragment length polymorphisms (RFLPs) at the Msc locus were identified in susceptible, kdr and super-kdr housefly strains. These RFLPs showed tight linkage to resistance in controlled crosses involving these strains, thus providing clear genetic evidence that kdr, and hence pyrethroid mode of action, is closely associated with the voltage-sensitive sodium channel.     

Identification and cloning of a key insecticide-metabolizing glutathione S-transferase (MdGST-6A) from a hyper insecticide-resistant strain of the housefly Musca domestica

Strains of the housefly, Musca domestica, highly resistant to organophosphate (OP) and other insecticides are known because they overproduce glutathione S-transferases (GSTs). Previous work has shown that overproduction in these strains involved numerous isozymes with glutathione conjugating activities (Pesticide Biochem. Physiol., 25 (1986) 169; Mol. General Genetics, 227 (1991) 355; J. Biol. Chem., 267 (1992) 1840; Mol. General Genetics, 245 (1994) 236; J. Mol. Evol., 43 (1996) 236). The current work describes the purification and identification of a M. domestica GST isozyme (pI 7.1) broadly specific for substrates from a housefly strain, Cornell-HR, that is highly resistant against OP-insecticides, and the isolation of two new MdGST genes using the antibody made against it. This isozyme, which was identified from amongst more than 20 isoelectric forms of GSTs of the same subunit size, was highly active for conjugating GSH to the model substrate 3,4-dichloronitrobenzne (DCNB). When expressed in Escherichia coli, one of the cloned GSTs, MdGST-6A, produces an enzyme that conjugates glutathione to the insecticides methyl parathion and lindane. On indication that it was the most active isozyme toward several xenobiotics among several MdGSTs tested, we advance the notion that MdGST-6A probably plays an important role in M. domestica Cornell-HR's resistance towards OP-insecticides. MdGST-6A and a second closely related one found in this work, MdGST-6B, are members of the traditional insect class I family (theta-class) and share the greatest homologies with a cluster of Drosophila GSTs on locus 55. In addition to having the unusually broad substrate specificity, the sequence of the new group of enzymes reveals that it has a highly diverged hydrophobic motif in its active site as compared to other class I GSTs from insects.     

The Hermes transposon of Musca domestica and its use as a mutagen of Schizosaccharomyces pombe

Transposon mutagenesis allows for the discovery and characterization of genes by creating mutations that can be easily mapped and sequenced. Moreover, this method allows for a relatively unbiased approach to isolating genes of interest. Recently, a system of transposon based mutagenesis for Schizosaccharomyces pombe became available. This mutagenesis relies on Hermes, a DNA transposon from the house fly that readily integrates into the chromosomes of S. pombe. The Hermes system is distinct from the retrotransposons of S. pombe because it efficiently integrates into open reading frames. To mutagenize S. pombe, cells are transformed with a plasmid that contains a drug resistance marker flanked by the terminal inverted repeats of Hermes. The Hermes transposase expressed from a second plasmid excises the resistance marker with the inverted repeats and inserts this DNA into chromosomal sites. After S. pombe with these two plasmids grow 25 generations, approximately 2% of the cells contain insertions. Of the cells with insertions, 68% contain single integration events. The protocols listed here provide the detailed information necessary to mutagenize a strain of interest, screen for specific phenotypes, and sequence the positions of insertion.     

苹果乙烯不敏感基因EIN2片段的克隆及序列分析

分别以苹果果实总DNA和cDNA为模板,采用PCR、RT-PCR方法扩增、克隆乙烯不敏感基因(ethyleneinsensitive 2,EIN2),并利用生物信息学方法分析其核苷酸序列和蛋白质结构。结果表明:(1)以DNA和cDNA为模板的扩增结果完全相同,扩增的EIN2基因片段为4 378bp,尚未发现有内含子,开放阅读框全长3 282bp,编码1 093个氨基酸;苹果EIN2相对分子质量为118.9kD,等电点为5.52,其蛋白可能为脂溶性疏水蛋白。(2)所克隆苹果EIN2基因编码的氨基酸序列与拟南芥(AAD41077.1)、碧桃(ACY78397.1)和葡萄(CAN66374.1)EIN2基因编码的氨基酸序列一致性分别为52%、79%、62%。(3)构建的EIN2基因进化树显示,拟南芥、小盐芥、甜瓜、杨毛果EIN2基因亲缘关系较近,聚为一类;葡萄为一类;蒺藜苜蓿为一类;碧桃、矮牵牛、西红柿聚为一类;苹果单独为一类。而且苹果EIN2基因与碧桃等同源基因的亲缘关系相对较近,与拟南芥、小盐芥同源基因的亲缘关系相对较远。     

Seasonal Prevalence ofEntomophthora muscaeand Introduction ofEntomophthora schizophorae(Zygomycotina: Entomophthorales) inMusca domestica(Diptera: Muscidae) Populations on California Dairies

Two entomopathogenic fungi in theEntomophthora muscaespecies complex that infect house flies were used in this study:E. muscaeFresenius (16–17 nuclei/conidium) occurred naturally at four southern California dairies, whileEntomophthora schizophoraeKeller and Wilding (4–8 nuclei/conidium) did not. During the first year of the study, onset of measurableE. muscaeinfections occurred between September and November but varied among sites. At least 20% of the flies at all four dairies were infected by November, and infection at one site exceeded 70%. During the fall epizootic period, infection levels were inversely related to temperature. Average weekly temperatures higher than 17–20°C and maximum daily temperatures higher than 26–28°C were statistically correlated with low infection levels. In the second year,E. schizophoraewas introduced by releasing diseased flies at two dairies (four times at one dairy and three times at the other).E. schizophoraewas recovered for a brief time in the house fly population after the first two releases at one site but not at the second site.