貉源犬瘟热病毒CDV-ZC株分离鉴定、全基因测序与分析

应用非洲绿猴肾细胞(Vero)从山东省诸城市疑似犬瘟热(canine distemper,CD)感染貉的肝脏、脾脏、肺脏等组织病料的研磨上清液中分离出1株病毒。分离株经Vero细胞传至第4代出现典型的细胞病变效应(CPE),经毒力测定、血清学、RT-PCR检测及测序、回归动物试验证明为犬瘟热病毒(Canine distemper virus,CDV),命名为CDV诸城分离株(CDV-ZC)。采用RT-PCR方法分段克隆分离株的全基因序列并测序,测序成功的各序列依次拼接得到全基因序列并进行序列比对。结果表明,CDV-ZC株全基因(KJ994343)与标准美洲型犬瘟热强毒株A75/17核苷酸同源性高达96.5%,而与疫苗株Onderstepoort、CDV3等亲缘关系较远,同源性为91.6%~92.0%。血凝素(H)基因序列分析表明,CDV-ZC株与国内野毒株LN(13)2、GP株等以及日本野毒株UENO、HAMA等共同归属于Asia-1型,在H蛋白信号淋巴细胞激活因子(SLAM)受体结合区即542~544位增加了1个潜在N-连接糖基化位点(N-X-S/T)。致病性强毒株CDV-ZC的成功分离及全基因序列测定加深了我们对当前中国CDV流行株遗传变异情况的了解,为CD的有效预防、诊断及控制提供理论依据。     

STRAIN ELASTOGRAPHY USING DOBUTAMINE‐INDUCED CAROTID ARTERY PULSATION IN CANINE THYROID GLAND

Thyroid disease is common in dogs and conventional ultrasonography is a standard diagnostic test for diagnosis and treatment planning. Strain elastography can provide additional information about tissue stiffness noninvasively after applying external or internal compression. However, natural carotid artery pulsations in the canine thyroid gland are too weak to maintain sufficient internal compression force. The objective of the present study was to describe the feasibility of strain elastography for evaluating the canine thyroid gland and the repeatability of dobutamine‐induced carotid artery pulsation as an internal compression method. In seven healthy Beagle dogs, strain on each thyroid lobe was induced by external compression using the ultrasound probe and internal compression using carotid artery pulsation after dobutamine infusion. The thyroid appeared homogeneously green and the subcutaneous fat superficial to the thyroid lobe appeared blue. Strain values and strain ratios did not differ among dogs or between the left and right lobes. Interobserver repeatability was excellent for both compression methods. Intraobserver repeatability of the strain ratio measured using the carotid artery pulsation method (intraclass coefficient correlation = 0.933) was higher than that measured using the external compression method (0.760). Mean strain values of thyroid lobes for the external compression method (142.93 ± 6.67) differed from the internal method (147.31 ± 8.24; P < 0.05). Strain ratios between the two methods did not differ. Strain elastography was feasible for estimating thyroid stiffness in dogs. Carotid artery pulsation induced by dobutamine infusion can be used for canine thyroid strain elastography with excellent repeatability.     

Effects of Different Substrates on Fermentation Parameters

Fermenting different raw materials (corn, soybean meal and cottonseed meal) with three kinds of strains (Lactobacillus, yeast and Bacillus subtilis) were employed to optimize the fermentation parameter with sensory evaluation, pH and amount of probiotics retention.The results showed that the time to reach stable pH of materials with different substrates was different.Cottonseed meal had the longest time to reach stable pH followed by soybean meal, while corn had the shortest time under the condition of same water content.Different substrates had no effect on initial pH of corn, soybean meal and cottonseed meal.The count of the probiotics was also influenced by different substrates.When pH reached steady state, corn had the largest number of the probiotics followed by soybean meal and cottonseed meal.The color and smell of three materials after fermented had distinctive features.But cottonseed meal had the highest viscosity followed by soybean meal and corn.Because of shorter time and more microorganisms, corn was better than soybean meal and cottonseed meal to be fermenting medium.     

人工组合复合菌系分解木质纤维素特征研究

为研究人工组合复合菌系降解木质纤维素的效果以及降解过程中微生物群落变化特征。首先对天然复合菌系PLC-8进行分离纯化,随后将筛选出的5株细菌和2株真菌进行组合,构建3 种简化的复合菌系,分别为F1、F2和F3。随后,设计秸秆降解试验,利用DGGE方法测定3种复合菌在分解纤维素过程中微生物群落的变化特征,并进行相关的理化分析。结果表明:人工组合复合菌系F3分解木质纤维素的能力最强,在分解木质纤维素30 d内秸秆平均减重率为39.80%,半纤维素下降52.64%,纤维素下降25.8%,木质素下降6.26%。在第15 天时纤维素内切酶、纤维素外切酶、滤纸酶、β-葡萄糖苷酶、木聚糖酶的活性已经达到整个分解过程的最高值,分别为0.09、0.08、0.09、0.07和0.11 U/mL。复合菌系F3能够自我调节pH,从而改变溶液的pH从微酸性至微碱性,F3的微生物群落呈现出真菌-细菌稳定共存的状态,并且能够长时间保持稳定。因此,人工组合复合菌系F3是稳定、高效的木质纤维素降解复合菌系。     

田间盆栽接种法鉴定若干向日葵品种(系)列当抗性

向日葵列当( Orobanche Cumana Wallr.) 是一种寄生性杂草,给向日葵生产造成严重威胁。为鉴定向日葵抗列当品种并为抗列当育种工作提供优异的种质资源,利用田间盆栽接种法鉴定向日葵新杂交组合以及恢复系、不育系对向日葵列当的抗性。于2019年对112个向日葵新品种(系),2020年对168个向日葵新品种(系)分别进行列当抗性鉴定。结果显示：2019年筛选出16个免疫,5个高抗的杂交组合;24个免疫、2个高抗的恢复系;7个免疫、2个高抗的不育系。2020年筛选出35个免疫、9个高抗的杂交组合;38个免疫,25个高抗的恢复系。2020年向日葵材料鉴定结果以免疫、高抗、中抗等抗性型为主,且均未发现高感材料,主要由于无论是杂交组合或恢复系,其亲本均来自之前几年鉴定得到抗性较好的种质资源。本单位经多年盆栽接种鉴定及自然发生列当的大田抗性验证,已获得若干抗性稳定的向日葵食葵、油葵品种(系)。     

猪肚菇菌株的选择及配方研究

【目的】筛选最适猪肚菇2号菌株和5号菌株的培养料配方,并选出栽培效益最高的菌株以备后期开发利用。【方法】利用上下限约束条件的混料试验方法设计26个培养料配方,与对照比较,筛选最佳培养料配方;栽培试验采用随机区组设计,四次重复,调查各处理的菌丝满袋时间、商品菇生物学效率、产量及效益。【结果】处理16配方最适合猪肚菇2号菌株,表现为菌丝35d满袋,两潮产量达163.9g/袋,商品菇生物学效率32.7%,每袋利润0.77元;处理9配方最适合5号菌株,表现为菌丝35d满袋,两潮产量达199.3g/袋,商品菇生物学效率39.9%,每袋利润1.55元。5号菌株的单袋产量及利润高于2号菌株。【结论】综合菌丝生长、产量及利润情况,在生产上选择猪肚菇5号菌株,其最适合的培养料配方为2.7%玉米芯+49%木屑+25.3%棉仔壳+2.0%玉米粉+18.2%麦皮+2.8%豆粕。     

扇贝新品系“紫海墨”F4的生长特征和生产性能分析

为考察扇贝新品系"紫海墨"的生产性能,为新品系选育效果提供依据,在同一海域养殖、相同管理模式和相同养殖周期下,对新品系"紫海墨"F4与墨西哥湾扇贝(Argopecten irradians concentricus)养殖群体的壳长、壳高、壳宽、体质量、闭壳肌质量和壳质量等7个性状及各阶段成活率进行生长特征和生产性能分析。结果表明,新品系"紫海墨"F4从10月13日养殖至翌年4月28日,壳长从1~1.5 mm长至50~60 mm的成品时,累积成活率为51.32%,显著大于墨西哥湾扇贝的29.18%(P<0.01);养殖周期内,两个养殖群体同一时期的壳长、壳高、壳宽和体质量也有极显著差异(P<0.01),新品系"紫海墨"F₄的生长速度高于墨西哥湾扇贝;经过6.5个月的养殖,新品系"紫海墨"F₄的壳长、壳高、壳宽、体质量和闭壳肌质量分别比墨西哥湾扇贝提高了19.50%、19.84%、16.23%、56.30%和20.20%。     

布鲁菌S2疫苗株免疫牛与牛种、羊种布鲁菌自然感染牛ELISA鉴别诊断方法的建立

为建立区分猪种布鲁菌S2疫苗株接种奶牛与布鲁菌自然感染奶牛,BLAST比对分析羊种、牛种、猪种、犬种、沙林鼠种和绵羊种6种布鲁菌基因序列,发现repA—related基因是猪种布鲁菌与牛种及羊种布鲁菌的差异基因。设计引物PCR扩增获得repA-related基因片段,克隆并原核表达得到了布鲁菌repA—related融合蛋白,以repArelated蛋白建立间接EI．IsA检测方法。用repA—related蛋白间接ELISA检测猪种s2疫苗株接种动物血清为阳性,检测牛种和羊种布鲁菌自然感染动物血清为阴性。repA—related蛋白间接EusA能从试管凝聚实验（SAT）及常规ELIsA检测阳性的奶牛血清样本中,区分出s2疫苗接种牛与牛种布鲁菌感染牛。     

四个平菇品种比较试验

为了筛选出适宜宁夏地区栽培的平菇品种,采用生料发酵法,利用日光温室袋栽技术,对4个供试品种菌丝生长情况、农艺性状、出菇能力及生物学效率等进行了比较研究。结果表明:印第安纳综合性状优良,菌丝长势强,子实体肉厚,菌盖颜色为灰黑色,短柄,生物学效率高,较受市场欢迎,可以在当地进行推广。     

利用响应面法优化全蚀病生防菌YB-81的发酵条件

为了优化全蚀病生防菌株YB-81的发酵培养基及培养条件,提高其对全蚀病的抑制效果,在单因素试验的基础上,利用响应面法(Response surface methodology)对小麦全蚀病生防菌株YB-81的发酵条件进行优化。生防菌株YB-81的最优发酵条件为:时间36h,温度36℃,初始pH值8.0,转速200 r·min-1,牛肉膏+葡萄糖(1∶1)0.53%,蛋白胨0.99%,氯化钠0.20%。在此培养条件下试验,其菌落总数为18.8亿个·mL-1,较基础培养基提高了72%,对全蚀病菌抑制率达到87.2%,较优化前提高了32.5%。生防菌YB-81的最优发酵培养条件的确定,可为该菌大规模生产应用提供理论基础。