基于转录组测序的大口黑鲈驯食相关SNP开发及其与生长性状的关联分析

大口黑鲈为肉食性鱼类,传统养殖过程中使用大量的冰鲜鱼为饵料,冰鲜鱼的使用不仅增加了养殖成本,而且多余的冰鲜鱼残饵还会给环境带来严重的污染。为了保护环境并降低养殖成本必须减少冰鲜鱼的使用量,本实验以选育适合人工配合饲料饲喂大口黑鲈为研究目标,用人工配合饲料作为饵料驯化大口黑鲈幼鱼,在驯食后14和16 d,测定其体质量、全长、体高等生长指标。同时在大口黑鲈转录组测序分析获得的SNP位点中,选择其所在序列的基因功能与能量代谢有相关性的7个SNP位点进行SNaPshot分型,用SPSS 19.0进行卡方分析标记与生长性状的相关性。结果发现,序列Unigene031044中的C1332G位点、Unigene085384中的A741G位点和Unigene022319中的A284G位点在体质量、全长及体高性状上存在显著差异。这3个SNP位点所在的基因经预测分别与雌二醇17β脱氢酶12B (hsd17b12b)基因、长链酰基辅酶A合成酶家族成员1(acsl1)基因和琥珀酸脱氢酶组装因子2(sdhaf2)基因具有很高的同源性,推测上述3个SNP位点的变异可能影响了基因表达产物对脂肪酸代谢或三羧酸循环的调节功能,与大口黑鲈对人工配合饲料的利用能力存在密切的相关性。本研究筛选得到的与驯食相关的SNP位点为大口黑鲈功能基因多态性的进一步研究,及加快大口黑鲈食性改良与驯化育种的遗传研究提供了科学的参考依据。     

野生动物用狂犬病口服疫苗复合免疫佐剂与诱饵的筛选及其对免疫效果的影响

本研究旨在筛选最佳的复合佐剂和口服疫苗诱饵组方,并将筛选出的复合佐剂口服免疫试验小鼠,检测免疫小鼠血清IgG和肠道IgA抗体效价。采用牛肉粉、鸡肉粉、鱼肉粉、奶粉、血粉为主要原料,以细菌脂多糖、氢氧化锌、枸杞多糖、甘露低聚糖为疫苗免疫佐剂,通过正交试验方法将复合免疫佐剂和口服疫苗诱饵与狂犬病SRV₉病毒混匀后口服免疫试验小鼠,并对免疫动物血清IgG抗体效价进行检测。优化的复合佐剂组方是细菌脂多糖100 μg、氢氧化锌0.8 mg、枸杞多糖50 mg、甘露低聚糖10 mg,免疫小鼠血清IgG抗体含量为3.24 mg/L,肠道IgA抗体含量为1.56 ng/mL;在疫苗诱饵的投喂试验中,狼、狐狸、犬、猫对鱼肉味诱饵的采食率最高,其次是血味和奶味诱饵;诱饵与狂犬病SRV₉病毒混匀后口服免疫小鼠28 d后,血清IgG平均值为1.20 U/L。本试验筛选出口服疫苗复合免疫佐剂,经筛选制备的鱼肉味诱饵具备高采食性,适合野生食肉类动物口服疫苗的制备。可增加狂犬病SRV₉病毒的免疫效应,为进一步研发重组减毒狂犬病口服疫苗提供了必要的技术支撑。     

Nicotinoid and pyrethroid insecticide resistance in houseflies (Diptera: Muscidae) collected from Florida dairies

BACKGROUND: The housefly, Musca domestica L., continues to be a major pest of confined livestock operations. Houseflies have developed resistance to most chemical classes, and new chemistries for use in animal agriculture are increasingly slow to emerge. Five adult housefly strains from four Florida dairy farms were evaluated for resistance to four insecticides (beta‐cyfluthrin, permethrin, imidacloprid and nithiazine). RESULTS: Significant levels of tolerance were found in most field strains to all insecticides, and in some cases substantial resistance was apparent (as deduced from comparison with prior published results). At the LC₉₀ level, greater than 20‐fold resistance was found in two of the fly strains for permethrin and one fly strain for imidacloprid. Beta‐cyfluthrin LC₉₀ resistance ratios exceeded tenfold resistance in three fly strains. The relatively underutilized insecticide nithiazine had the lowest resistance ratios; however, fourfold LC₉₀ resistance was observed in one southern Florida fly strain. Farm insecticide use and its impact on resistance selection in Florida housefly populations are discussed. CONCLUSION: Housefly resistance to pyrethroids is widespread in Florida. Imidacloprid resistance is emerging, and tolerance was observed to both imidacloprid and nithiazine. If these insecticides are to retain efficacy, producer use must be restrained. Copyright © 2009 Society of Chemical Industry     

A new fire ant (Hymenoptera: Formicidae) bait base carrier for moist conditions

BACKGROUND: A new water‐resistant fire ant bait (T‐bait; cypermethrin 0.128%) consisting of dried distillers grains with solubles (DDGS) as a carrier was developed and evaluated against a standard commercial bait (Advion^®; indoxacarb 0.045%) under both laboratory and field conditions. RESULTS: When applying the normal T‐bait or Advion^® in the laboratory, 100% of Solenopsis invicta Buren worker ants were killed within 4 days. However, when the T‐bait and Advion^® were wetted, 70.6 and 39.7% of the ants were killed respectively. Under field conditions, dry T‐bait and dry Advion^® had almost the same efficacy against ant colonies. However, when T‐bait and Advion^® came in contact with water, the former's ability to kill S. invicta colonies in the field was only marginally reduced, while Advion^® lost virtually all of its activity. In addition, DDGS was also shown to be compatible with a number of other insecticides, such as d‐allethrin, permethrin and pyrethrin. CONCLUSION: Based on its properties of remaining attractive to the fire ants when wetted, combined with its ant‐killing abilities both in the laboratory and in the field, T‐bait is an efficient fire ant bait, especially under moist conditions. Copyright © 2010 Society of Chemical Industry     

4种饵料对玉足海参幼体生长和发育的影响

为了解单一饵料或混合投喂对玉足海参(Holothuria leucospilota)幼体生长的影响,采用2种单胞藻(牟氏角毛藻、球等鞭金藻)和2种酵母(酿酒酵母、海洋红酵母)单一或组合搭配投喂玉足海参耳状幼体,测定并比较海参幼体的体长、胃宽、水体腔长、变态成活率等指标。试验结果显示：单一饵料组中,牟氏角毛藻对玉足海参幼体体长的生长及变态存活最有利,而球等鞭金藻对其脏器(胃、水体腔)的发育最有利;综合所有处理组的投喂效果看,混合投喂组比单一饵料组更有利于玉足海参幼体的生长和存活,其中酿酒酵母+海洋红酵母的混合投喂组合得到了最高的体长、胃宽和水体腔长特定生长率,牟氏角毛藻+海洋红酵母投喂组的变态成活率最高,变态发育速度最快。结果表明,在玉足海参幼体培养早期适合以混合酵母为主投喂,后期则可较多投喂牟氏角毛藻和海洋红酵母。     

Effect of dietary carbohydrate to lipid ratios on growth,digestive enzyme and blood metabolites of juvenile Brazilian sardines,Sardinella brasiliensis (Steindachner, 1879)

The limited availability of live bait for capturing skipjack tuna, Katsuwonus pelamis, is a bottleneck to increasing tuna production in many parts of the world. Therefore, a nutrition trial was performed to contribute to the production of the Brazilian sardine, Sardinella brasiliensis, for use as live bait. This study determined the best dietary carbohydrate to lipid ratio (CHO:L) for juvenile Brazilian sardines based on growth performance, feed utilisation, body composition, blood metabolites and digestive enzyme activity. Six isoenergetic and isonitrogenous diets were formulated with increased CHO:L ratios (2.05, 3.41, 4.15, 5.11, 5.80 and 6.72). Each diet was randomly assigned to triplicate groups of 100 fish with mean initial body weight of 2.97 ± 0.51 g, which were fed four times a day to apparent satiation. Survival was not affected by differences in diet, however, a low CHO:L ratio stimulated growth. Juveniles fed with a rich‐carbohydrate diet inhibit feed intake and protein intake. Body lipid increased as dietary lipid increased and was inversely correlated to body moisture. The diets did not affect the juvenile's blood metabolites. Alkaline and acid protease activities were not significantly different, but lipase and amylase responded positively to the dietary lipids and carbohydrates. Using segmented regression, the optimum CHO:L ratio for maximum weight gain of juvenile Brazilian sardines was estimated to be 3.41, which contain approximately 300 g kg^?1 carbohydrate and 88 g kg^?1 lipid.     

酵母双杂交技术筛选与绵羊微管解聚蛋白相互作用的蛋白

采用TRIzol法提取绵羊卵巢组织总RNA,运用RT–PCR技术扩增绵羊微管解聚蛋白基因(STMN1)的编码区序列;通过同源重组技术构建重组诱饵表达载体p GBKT7–STMN1,鉴定该诱饵蛋白在酵母双杂交系统中的自激活活性、细胞毒性和表达情况;利用诱饵质粒p GBKT7–STMN1从绵羊卵巢组织细胞的酵母双杂交cDNA文库筛选互作的宿主蛋白。结果显示：成功构建重组诱饵质粒p GBKT7–STMN1,并对酵母细胞无自激活活性和细胞毒性作用,且能在酵母细胞中正常表达;筛选获得12个与STMN1相互作用的克隆,经测序比对分析和点对点验证,得到CREB和FOXM1共2个互作的宿主蛋白,这2个蛋白可能参与调控细胞的增殖、周期、凋亡等过程。     

4种饵料微藻对水产养殖废水的净化效果研究

为探究饵料微藻在养殖废水中的生长能力以及去除氮、磷的效果,选用牟氏角毛藻(Chaetoceros muelleri)、青岛大扁藻(Platymonas helgolandica)、绿色巴夫藻(Pavlova viridis)和海水小球藻(Chlorella vulgaris)等4种微藻,在室内光照培养系统中进行了微藻净化养殖废水试验,试验时长为11 d。结果表明,绿色巴夫藻对NH_4~+-N和NO_2~--N去除效果最佳,最大去除率分别为88.35%和97.43%;海水小球藻对废水中PO_4~(3-)-P的去除率最高,为93.29%;就生长情况而言,绿色巴夫藻平均相对生长率最高,达到0.239 cells·mL~(-1)·d~(-1)。因此,在本试验条件下,可认为绿色巴夫藻是治理养殖废水的优势藻种。     

家白蚁高效诱饵的研制及诱效试验

为了研制白蚁高效诱饵剂，在室内用胶卷盒装各种锯木粉及添加剂，置于干燥器内，测定家白蚁对各种饵料的趋性，在室外把饵料盒埋在诱集箱外，42d后称重.室内试验松木粉及杉木粉中加入添加剂后,对白蚁的引诱力提高7.9及24.9倍；室外试验饵料中加入添加剂后，可提高白蚁对其取食量1.22～6.48倍.含萜类物过多的松、杉木的锯木粉，经烘、煮等处理后也可提高白蚁对它们的喜食程度.     

Assessing insecticide resistance and aversion to methomyl-treated toxic baits in Musca domestica L (Diptera: Muscidae) populations in southern California

Progeny of house flies (Musca domestica L) from ten California poultry operations, three dairies and one horse-riding facility were tested for methomyl- and muscalure-treated bait resistance using up to three different assays: a topical assay, a no-choice feeding assay and a choice feeding assay. LD50 resistance factors from the topical assay, compared with a locally-derived susceptible colony, ranged between 1 and 4. LC50 resistance factors from the no-choice feeding assay ranged mostly between 2 and 5, with one value of 18. Measurable LT50 resistance ratios for female flies in the choice feeding assay ranged from 43 to 159; two populations had <10% mortality at 48h and could not be measured. LT50 resistance ratios for male flies in the choice feeding assay ranged from 26 to 96, and one population was too resistant to measure. A behavioral assay tested the feeding preference of male and female flies provided a dish of sugar and a dish of methomyl- and muscalure-treated bait. Of eight strains tested, females from seven strains and males from six strains showed significant preference for sugar over bait. Behavioral factors appear to be important in the severe resistance of house flies to baits in California.