CRISPR/cas系统及其在家禽上的应用研究进展

CRISPR/cas是一种获得性免疫防御系统,广泛存在于细菌和古细菌中。Strep tococcus pyogenes cas9(Spcas9)作为目前研究最多、最清楚的效应蛋白,因其酶活性高和靶点广而被应用于生命科学各个领域,但Spcas9分子量大、脱靶率高等缺点在一定程度上限制了其应用。随着CRISPR系统技术被深入开发,一些新效应蛋白被发现,本文就CRISPR/cas系统分类、原理、新效应蛋白发现及cas9(S.Pyogenes)多种变构体技术开发在及家禽上的应用进行综述,为相关领域的研究提供参考。     

桑蚕卵中酯酶A_4的研究

取产后两天内25℃保存的桑蚕卵为材料,精制酯酶A_4。将滞育卵中精制的酶液置5℃下冷藏,冷藏初期酶活性极低,大致在12～13日达最大值。以后迅速降低,体外保存的温度条件与酯酶A_4活性有关,较高的保存温度能促使酶活性峰提早出现:而低温保护能延缓酶的活化进程。酯酶A_4精制过程中,通过Sephadex G-25凝胶过滤,将大部分物质分段与酶分离排除,在5℃条件下,酶活性峰出现以前的任何时候加入排除分段物质,都可以停止酶的活性化,只有当重新除去所加的排除分段物质时,酶的活性化进程才得以继续。     

春制越年家蚕卵乙胚子冷藏方法研究

研究结果表明，春制越年家蚕卵丝乙胚子冷藏处理后，其实用孵化率〉９２．３％蚁蚕绝食致半数时数≥１１０ｈ，不良卵发生率〈１０％，发病率〈８．６％，茧质优良，产量平稳，在有效冷藏期限内随时出库催青饲育均安全可靠。     

鸡蛋消毒方法的对比研究

采用5种消毒方法对鸡蛋蛋壳表面细菌进行杀菌效果试验,结果表明,各处理组之间抑菌效果差异显著(P＜0.05),杀菌效果优劣依次为开水浸烫＞甲醛熏蒸＞过氧乙酸熏蒸＞高锰酸钾浸泡＞过氧乙酸浸泡,开水浸烫操作技术难度较大,容易引起蛋壳破裂,适宜实际生产的消毒方法为过氧乙酸熏蒸和甲醛熏蒸.将消毒处理后的鸡蛋在温度25℃,相对湿度60%～80%贮存20天,开水浸烫处理组鸡蛋仍保持较鲜的品质,其余各组鸡蛋品质较差.结果表明,常规消毒方法(除开水浸烫法)只能抑制蛋壳表面微生物的生长繁殖,不能延缓鸡蛋品质下降达到保鲜鸡蛋的作用.     

Effects of in ovo injection of chrysin,quercetin and ascorbic acid on hatchability,somatic attributes,hepatic oxidative status and early post‐hatch performance of broiler chicks

An experiment was conducted to evaluate the effects of in ovo injection of chrysin, quercetin and ascorbic acid on hatchability, somatic attributes, hepatic antioxidant status and early post‐hatch growth performance of broiler chicks. Four hundred and eighty embryonated broiler breeder eggs containing live 18‐day‐old embryos were divided into six groups of 80 eggs each. One group remained intact and served as a control group (i), whereas the other five groups were injected with the prepared injection solutions as follows: (ii) 0.05 ml distilled water; (iii) 0.05 ml distilled water containing 6 mg ascorbic acid; (iv) 0.05 ml dimethyl sulfoxide (DMSO); (v) 0.05 ml DMSO containing 4.5 mg quercetin; and (vi) 0.05 ml DMSO containing 4.5 mg chrysin. The hatchability rate, hatching weight, residual yolk sac weight, yolk sac‐free body weight, liver weight, hepatic glutathione peroxidase and total superoxide dismutase activities, as well as malondialdehyde concentrations, were not affected by the injected solutions. There were no differences between chicks hatched from the control and in ovo injected eggs in weight gain, feed intake and feed conversion ratio from 0 to 11 days of age. However, the specific contrast performed between the in ovo injected groups and intact eggs revealed that in ovo injection significantly increased hatchability rate (p = .0493). This finding also implies that our injection procedure was harmless. In conclusion, the intra‐egg injection of chrysin, quercetin or ascorbic acid at the injection rates used in this study did not have a significant effect on hatchability, somatic characteristics, early growth performance and hepatic antioxidant status of broiler chicks. However, the overall hatchability was higher in the in ovo injected eggs as compared to non‐injected ones. These findings also confirmed the harmlessness of the procedure developed for in ovo injection in this study.     

皖南黄肉种鸡种蛋中禽白血病病毒的感染状态检测

本研究首次通过检测种蛋中禽白血病病毒(ALV)评价了皖南黄父母代肉种鸡的ALV感染状态.收集该鸡群的120枚鸡蛋,取40枚鸡蛋孵化至9～11 d后分别制备CEF,培养10 d,取细胞上清用ELISA试剂盒检测ALV p27抗原.另取80枚鸡蛋卵白直接检测p27抗原,同时分别将80枚鸡蛋卵白接种CEF(DF1),培养10 d后取细胞上清检测p27抗原.比较p27阳性检出率的结果表明,受精蛋孵化9～11 d后制备CEF,再培养10 d的细胞上清检出率(10/36)高于直接检测卵白(17/80)与卵白接种CEF(DF1)的细胞上清(7/80).将经DF1培养后p27抗原阳性样品,用针对ALV-J的单抗JE9做IFA,ALV-J的阳性率为6/7.检测该80枚鸡蛋的卵黄抗体,ALV-J、ALV-AB的抗体阳性率分别为18/80、1/80.结果表明,该皖南黄父母代肉种鸡群存在的外源性ALV感染主要为ALV-J.该研究为ALV的净化提供了可行性检测方法.     

Littoral spawning habitats of three southern Arctic charr (Salvelinus alpinus L.) populations

Abstract Arctic charr populations in southern latitudes are nonmigratory, with all life‐stages limited to freshwater lakes and in‐ or out‐flowing tributaries. Although many of these populations are reported to also spawn in lake littorals, little is known about the physical characteristics of putative spawning grounds. A total of 23 discrete spawning sites within three Irish lakes were located by fyke netting of spawning adults and snorkelling in littoral habitats. Spawning sites were found to be long, narrow strips running parallel to the shore at a maximum depth of 124 cm. Spawning sites were limited to areas of coarse mineral substrate with an adequate (c. 8 cm) depth of clean interstitial spaces. In individual lakes, combined areas of spawning sites made up 0.4–0.7% of available littoral. Egg densities varied considerably between sites (33–900·eggs m^?2) and were significantly correlated with gradient and width of spawning sites. No evidence of redd digging was found. The shallow, localised and restricted nature of spawning grounds makes such populations vulnerable to anthropogenically induced postoviposition changes in surface water level, eutrophication processes such as increased lake sedimentation and elevated nutrient status.     

Nutritional significance of amino acids,vitamins and minerals as nutraceuticals in poultry production and health – a comprehensive review

Nutraceuticals have gained immense importance in poultry science recently considering the nutritional and beneficial health effects of their constituents. Besides providing nutritional requirements to birds, nutraceuticals have beneficial pharmacological effects, for example, they help in establishing normal physiological health status, prevent diseases and thereby improve production performance. Nutraceuticals include amino acids, vitamins, minerals, enzymes, etc. which are important for preventing oxidative stress, regulating the immune response and maintaining normal physiological, biochemical and homeostatic mechanisms. Nutraceuticals help in supplying nutrients in balanced amounts for supporting the optimal growth performance in modern poultry flocks, and as a dietary supplement can reduce the use of antibiotics. The application of antibiotic growth enhancers in poultry leads to the propagation of antibiotic-resistant microbes and drug residues; therefore, they have been restricted in many countries. Thus, there is a demand for natural feed additives that lead to the same growth enhancement without affecting the health. Nutraceuticals substances have an essential role in the development of the animals’ normal physiological functions and in protecting them against infectious diseases. In this review, the uses of amino acids, vitamins and minerals as well as their mode of action in growth promotion and elevation of immune system are discussed.     

Residue Detection of Tetracyclines in Eggs by High Performance Liquid Chromatography

The study was aimed to establish the method of simultaneous detection of four kinds of tetracyclines,including oxytraoycline,tetracycline,chlortetracycline and doxyeycline in eggs by high performance liquid chromatography (HPLC).The eggs samples were detected with HPLC after through the procedure of extraction of 0.1 mol/L Na₂EDTA-Mcllvaine buffered solution,purification of Oasis MAX solid-phase extraction column,gradient elution of taking acetonitrile-0.01 mol/mL trifluoroacetic acid as mobile phase,separation with reverse liquid chromatography.The results showed that the calibration curves of four kinds of tetracyclines were linear correlated in the concentration range of 30 to 1 000 ng/g which the limits of detection was 10 ng/g and the limit of quantification was 50 ng/g.The average recoveries were 67.3% to 81.4%,relative standard deviations of intraday was 2.7% to 14.3%,relative standard deviations of inter-day was 0.9% to 5.0%.This mothod established in this study could simultaneously detect oxytetracycline,tetracycline,chlortetracycline and doxycycline drug residues in egg and it had high sensitivity,accuracy and precision.The method was rapid and simple to operate.