甘蔗黄叶病毒与花叶病毒CP基因RNAi载体构建与转化甘蔗研究

甘蔗黄叶病和甘蔗花叶病是我国甘蔗最主要的病毒病,感病品种产量下降和品质变劣,传统方法难以防治。RNA干扰技术使培育抗病毒甘蔗品种成为可能。本研究根据甘蔗黄叶病毒和侵染甘蔗的高粱花叶病毒结构与功能特性,广泛收集不同来源病毒分离物CP基因序列,经过比对选取保守序列作为干扰序列。通过在序列两端添加酶切位点,合成并连接成双价甘蔗黄叶病和花叶病毒干扰序列,然后构建到中间载体p HANNIBAL上,形成双价RNAi干扰结构,最后插入到p ART27上形成干扰表达载体。利用基因枪轰击甘蔗品种福农15号愈伤组织进行转化,经G418筛选获得抗性再生植株。通过提取DNA和RNA分析,证实双价RNAi干扰结构不仅以不同拷贝整合到甘蔗基因组中,而且得到了转录表达。     

Optimum Level of Dietary n‐3 Highly Unsaturated Fatty Acids for Juvenile Sea Cucumber,Apostichopus japonicus

A feeding trial was conducted to estimate the optimum level of dietary n‐3 highly unsaturated fatty acids (HUFAs) for juvenile sea cucumber, Apostichopus japonicas, based on growth performance and fatty acid compositions. Diets with five n‐3 HUFAs levels (0.15, 0.22, 0.33, 0.38, and 0.46%) were fed to sea cucumber juveniles (1.97 ± 0.01 g) once a day for 60 d. The sea cucumbers fed diets containing 0.22% n‐3 HUFAs showed significantly (P < 0.05) higher body weight gain, feed efficiency, and protein efficiency ratio than the sea cucumbers fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.33, 0.38, and 0.46% n‐3 HUFAs. The sea cucumbers fed diets containing 0.46% n‐3 HUFAs showed significantly (P < 0.05) higher eicosapentaenoic acid and saturated fatty acid than the sea cucumber fed diets containing 0.15% n‐3 HUFAs, but not significantly different (P > 0.05) from those of sea cucumbers fed diets containing 0.22, 0.33, and 0.38% n‐3 HUFAs. The results of growth performance and n‐3 HUFA compositions of body wall indicated that the optimum level of dietary n‐3 HUFAs for juvenile sea cucumber is between 0.22 and 0.46%.     

益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究

本试验旨在研究益生菌大肠杆菌Nissle 1917(Ec N)抗逆性能、猪肠上皮细胞黏附率及抑菌效果。采用体外法对Ec N进行生长曲线绘制和耐酸、耐胆盐、耐热性能的测定;以猪肠上皮细胞IPEC-J2细胞为体外细胞模型,考察了Ec N对该细胞的黏附率以及对致病菌大肠杆菌K88的黏附抑制率;同时通过蛋白质印迹法检测了Ec N对IPEC-J2细胞β-防御素-2和Toll样受体4的水平的影响。结果表明:1)Ec N对高酸、高胆盐和高温环境具有一定耐受能力。2)Ec N对IPEC-J2细胞的黏附作用以对数期最佳,黏附率达33.96%,显著高于迟缓期、稳定期和衰亡期(P0.05)。3)Ec N对致病菌大肠杆菌K88具有良好的抑制效果,黏附抑制率达87.84%。4)Ec N还能上调IPEC-J2细胞β-防御素-2和Toll样受体4水平。结果提示,益生菌Ec N具有较好的抗逆性能,能够良好地黏附猪肠上皮细胞,对致病菌大肠杆菌K88具有良好的抑制作用。     

Effect of ammonia and nitrite on vigour,survival rate,moulting rate of the blue swimming crab <Emphasis Type="Italic">Portunus pelagicus</Emphasis> zoea

The effects of ammonia and nitrite on vigour, survival rate, moulting rate of zoea of blue swimming crab, Portunus pelagicus, were studied. A total of five nitrite-N treatments (26.67, 53.34, 106.68, 213.36, 426.72 mg/l) and a control (no nitrite-N added) were set up for the acute nitrite-N toxicity experiment; a total of five ammonia-N treatments (8.43, 16.86, 33.72, 67.44, 134.88 mg/l) and a control (no ammonia-N added) were set up for the acute ammonia-N toxicity experiment. The results showed that the vigour, survival rate and moulting rate of zoea of the blue swimming crabs exposed to over 53.34 mg/l were significantly different (P < 0.05) from the control group. The zoea LC₅₀ values (mg/l) of nitrite-N were 179.47, 76.56, 66.70, 37.49, 25.01, 25.35, 25.34 mg/l for 24, 36, 48, 60, 72, 84, 96 h, respectively. The vigour, survival rate and moulting rate of zoea of the blue swimming crabs exposed to over 16.86 mg/l were significantly different (P < 0.05) from the control group. The zoea LC₅₀ values (mg/l) of ammonia-N were 51.04, 39.62, 38.72, 24.43, 16.90, 13.42, 11.16 mg/l for 24, 36, 48, 60, 72, 84, 96 h, respectively. The zoeae are highly sensitive to ammonia and nitrite, and the toxicity of ammonia and nitrite on Portunus pelagicus decrease with development of this crab.     

小型猪乳化异氟醚全身麻醉效果监测

为探讨乳化异氟醚进行小型猪全身麻醉麻醉效果,选取14头巴马猪以2.80 mL.kg-1.h-1进行静脉麻醉,在麻醉过程中对生理反射、镇痛、镇静、肌松,呼吸和循环系统进行监测。结果表明,乳化异氟醚可以使小型猪维持良好外科麻醉状态,麻醉过程中小型猪一般生理指标、循环系统指标未发生显著变化(P>0.05),呼吸系统指标与正常比较有显著性改变(P<0.05或P<0.01),但均在安全范围内,小型猪麻醉过程无异常表现。乳化异氟醚能满足临床手术麻醉需要,可以作为小型猪全身麻醉药物推广应用。     

鹦哥岭自然保护区海南锯腿树蛙种群分布以及繁殖生物学研究

2011年2月~2012年7月,对海南鹦哥岭自然保护区内的海南锯腿树蛙(Rhacophorus hainanus)种群分布进行了调查,用野外观察与室内饲养的方法对其繁殖生物学进行了初步研究。结果表明:海南锯腿树蛙分布在鹦哥岭自然保护区内海拔800m以上的溪流边或是季节性水潭附近;该蛙具有明显的性二型特点;繁殖季节为每年的3月至6月,繁殖地为季节性水潭,繁殖包括鸣叫求偶、抱对、产卵三个过程,每次产卵量为126±77枚,蛙卵约7d孵化为蝌蚪,57d蝌蚪变态成为幼蛙。该蛙唇齿式为I:4+4/1+1:II,与云南部分蝌蚪标本以及越南采集的蝌蚪标本有差异。     

Molecular identification and relative abundance of microorganisms in douchi koji and salted egg white sufu during processing

A novel animal protein-based douchi koji-inoculated steamed salted egg white sufu (SEWS) has been developed. This study determined the relative abundance of microorganisms in the douchi koji and semi-finished (5-day fermentation) and finished (5-day fermentation and 14-day ripening) SEWS by using 16S and 18S ribosomal DNA and gene-cloning methods. The results revealed that Bacillus spp. and Aspergillus oryzae were dominant in the douchi koji. In the semi-finished SEWS, the percentages of Bacillus amyloliquefaciens and Bacillus subtilis were considerably lower, whereas those of Enterococcus and Staphylococcus were substantially higher. In the finished SEWS, Bacillus spp. became dominant again and A. oryzae was the only fungus detected. In conclusion, by using molecular techniques, microbial population dynamics in SEWS can be evaluated. During processing, the relative abundance of microorganisms in SEWS changed and Bacillus spp. and A. oryzae remained dominant. This study provides crucial information for designing starter cultures for producing SEWS.     

Association between weight change during initial chemotherapy and clinical outcome in dogs with multicentric lymphoma

The majority of the known prognostic factors in dogs with lymphoma have been evaluated before treatment commences or at the time of diagnosis. Prognostic factors evaluated during the initial phase of treatment are less described but may provide important clinical information. In this retrospective study, 82 canine lymphoma patients were categorized according to the weight change between diagnosis and after 5 weeks of chemotherapy. Dogs that gained greater than 5% or lost greater than 5% of initial body weight were categorized as increased‐ or decreased‐weight groups, respectively. Those in which weight changed less than 5% were categorized as the maintained‐weight group. The median progression‐free survival (PFS) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 226, 256 and 129 days, respectively. The decreased‐weight group had significantly shorter PFS than the increased and maintained groups (P = .023, P = .003, respectively). The median survival time (ST) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 320, 339 and 222 days, respectively. There was no significant difference in ST among the three groups (P = .128). In Cox‐regression results, weight change group and initial body weight were significant risk factors associated to PFS (P = .007, P = .001, respectively) while only patient's initial body weight was a significant risk factor to ST (P = .013). In conclusion, evaluation of initial body weight and weight changes over time can provide valuable information regarding PFS and ST in dogs with multicentric lymphoma.     

射频识别技术在畜禽养殖中的应用与展望

随着畜禽规模养殖的发展,高度集约化养殖模式将影响畜禽的健康状况、疫病防控和福利水平。一种无接触、无应激的射频识别（Radio Frequency Identifi cation,RFID）技术应运而生,该技术可以在无损状态下采集畜禽的行为、生理生化和养殖环境指标,从而为评估集约化养殖的效果和影响提供基础数据。文章讨论了三种常见类型的RFID系统在畜禽养殖中的应用,系统地比较了各类系统的优缺点及适宜应用场景,进一步提出RFID在畜禽养殖中的未来应用方向,以期为深化畜禽信息采集的研究提供参考。     

Comparison of efficacy and toxicity of doxorubicin and mitoxantrone in combination chemotherapy for canine lymphoma

Forty-four dogs with multicentric lymphoma were treated using a cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) induction protocol or treated using a cyclophosphamide, mitoxantrone, vincristine, and prednisolone (CMOP) induction protocol. There was no statistical difference in signalment and the presence of historical negative prognostic factors between the groups. The median progression-free survival (PFS) in the CHOP and CMOP groups were 222 d and 162 d, respectively (P = 0.75). The median survival time (MST) of dogs in CHOP and CMOP groups were 318 d and 242 d, respectively (P = 0.63). Anorexia and diarrhea episodes were significantly higher in the CHOP group than in the CMOP group (P = 0.02 and P = 0.01, respectively). These results suggest that the CMOP protocol provides similar PFS, MST and causes fewer side effects compared to the CHOP protocol. Therefore, the CMOP protocol may be another treatment choice for canine multicentric lymphoma.