The pharmacokinetics of clomipramine and desmethylclomipramine in dogs: parameter estimates following a single oral dose and 28 consecutive daily oral doses of clomipramine

Clomipramine is a tricyclic antidepressant that has been recommended for the treatment of canine compulsive disorder. The pharmacokinetics of clomipramine in dogs have not been reported. This study describes the pharmacokinetics of clomipramine and its active metabolite, desmethylclomipramine, in six male dogs. Serial blood samples were collected following both a single oral dose of clomipramine (3 mg/kg) and 28 consecutive daily oral doses (3 mg/kg q 24 h). In addition, 'peak' and 'trough' samples were taken throughout the 28-day dosing period. Plasma was assayed for total (free and protein-bound) clomipramine and desmethylclomipramine, using gas-chromatography with mass spectrometric detection. Various pharmacokinetic parameters were then determined. Following a single dose of clomipramine, time of maximum plasma concentration ( t _max) of clomipramine was 0.75–3.1 h, maximum plasma concentration ( C _max) was 16–310 ng/mL and elimination half-life ( t _1/2el) was 1.2–16 h; t _max of desmethylclomipramine was 1.4–8.8 h, C _max was 21–134 ng/mL and t _1/2el was 1.2–2.3 h. Following multiple dosing, there was a numeric increase in these parameters; t _max of clomipramine was 3–8 h, C _max was 43–222 ng/mL and t _1/2el was 1.2–16 h; t _max of desmethylclomipramine was 1.4–8.8 h, C _max was 21–134 ng/mL and t _1/2el was 1.2–2.3 h. Clinically significant differences between dogs and humans in the pharmacokinetics of oral clomipramine are discussed.     

一株H9N2亚型禽流感病毒分离株的生物学特性研究及其全基因组序列分析

本试验对1株2010年从广东省分离的H9N2亚型禽流感病毒A/chicken/Guangdong/QY/2010(H9N2)的生物学特性进行研究并对其全基因组序列进行分析。结果显示,该毒株的鸡胚半数感染量 EID₅₀为10^－9/0.1 mL,鸡胚最小致死量的平均死亡时间MDT为104 h,脑内致病指数ICPI值为0.51,静脉致病指数IVPI为0。用RT-PCR方法扩增病毒的基因组各片段,将扩增片段进行克隆、测序并进行序列分析。结果显示,该毒株的HA基因与Ck/HK/G9/97和Dk/HK/Y280/97在同一分支上,HA的裂解位点为PARSSR↓GLF,有8个潜在糖基化位点,226位氨基酸残基为L,较保守的202位受体结合位点由L突变为P,该毒株的HA和NA核苷酸序列与Dk/HK/Y280/97同源性较高,NS、PA、PB1的核苷酸序列均与Ck/SH/F/98同源性较高,NP基因与A/VN/ 1203/04(H5N1)的核苷酸序列同源性为95.3%。     

伪狂犬病病毒在NF-κB家族p65基因敲除细胞系的复制规律研究

试验旨在研究伪狂犬病病毒（PRV）在NF-κB家族p65基因敲除细胞系中的复制规律。利用慢病毒介导的CRISPR/Cas9基因定点修饰技术构建猪肺泡巨噬细胞（3D4/21）p65基因稳定敲除细胞系。通过构建p65-sgRNA重组质粒,转染至HEK293T/17细胞,收取慢病毒,感染3D4/21细胞后利用嘌呤霉素筛选获得多克隆细胞系,T7核酸酶检测敲除效率,再通过有限稀释法获得3D4/21-p65^-/-的稳定细胞系。CCK-8试剂盒检测3D4/21细胞中敲除p65基因后对细胞增殖的影响;流式细胞术检测PRV-GFP感染3D4/21及3D4/21-p65^-/-细胞后病毒增殖的差异;实时定量PCR检测PRV感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、TK基因mRNA表达水平及PRV感染细胞诱导的IL-1β和IL-6基因mRNA水平表达的变化;Western blotting检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后PRV gB、gE蛋白的表达;滴度测定检测PRV-QXX感染3D4/21及3D4/21-p65^-/-细胞后子代病毒滴度。结果表明,sgRNA2和sgRNA3的基因编辑效率较高,对其进行克隆化培养进而获得敲除p65基因的稳定表达细胞系;CCK-8试剂盒检测细胞活力表明,p65基因敲除对细胞活力无影响;流式细胞仪检测表明,同一时间点PRV-GFP在3D4/21-p65^-/-中的增殖显著高于对照细胞;实时荧光定量PCR表明在3D4/21细胞中敲除p65基因促进了PRV gB、TK基因的mRNA表达水平,而抑制了IL-1β、IL-6基因的mRNA表达;Western blotting结果表明,在3D4/21细胞中敲除p65基因促进了PRV gB、gE蛋白的表达;滴度测定结果表明,同一时间点PRV-QXX在3D4/21-p65^-/-细胞中子代病毒的复制显著高于对照细胞。以上结果均表明,p65基因敲除可促进PRV在3D4/21细胞中复制。     

Exon skipping of exonuclease 1 in MRL/MpJ mice is caused by a nucleotide substitution of the branchpoint sequence in intron eight

In MRL/MpJ mice, there is a genetic mutation of exonuclease 1 (Exo1), in which the exon 9 is sometimes deleted. In the present study, to check the generation of the spliced exons, exon 8-intron 8-exon 9 (pCX/Ex/EIE/B and pCX/Ex/EIE/M) plasmids were temporally transfected in vitro into BALB 3T3 cells, and RT-PCR using appropriate primer pair was carried out 1 day after transfection. In these constructions, pCX/Ex/EIE/B was derived from genomic sequence of C57BL/6 mice, and pCX/Ex/EIE/M was from MRL/MpJ. A spliced band was detected in pCX/Ex/EIE/B, but was present little or very weakly in pCX/Ex/EIE/M. Next, the same spliced band was demonstrated in the pCX/Ex/EIE/M(T) plasmid, in which the branchpoint sequence (BPS) of pCX/Ex/EIE/M including the exon 9 was changed into that of pCX/Ex/EIE/B. The splicing did not occur in the dell1/B mutant, in which 1960 nucleotides of the intron 8 were deleted, whereas it was detected in the del2/B plasmid deleted 1036 nucleotides in its middle region. These results suggest that the nucleotide T to A mutation of the BPS in the intron 8 is at least a sufficient for generation of splice variants (tr-1 and tr-2 Exo1).     

Pharmacokinetics of oxytetracycline hydrochloride in rabbits

Pharmacokinetics of oxytetracycline HCl (OTC) was studied in rabbits. After 10 mg of OTC/kg of body weight was administered IV, the distribution half-life was 0.06 hour, terminal half-life was 1.32 hours, volume of distribution area was 0.861 L/kg, and total body clearance was 0.434 L/kg/h. After 10 mg of OTC/kg was given IM, the absorption half-life was 2.09 hours, extent of absorption was 71.4%, and total body clearance of the absorbed fraction was 0.576 L/kg/h. Based on these kinetic data, a dosage of 15 mg of OTC/kg, every 8 hours was developed. This dose given IM for 7 consecutive days resulted in observed steady-state maximum and minimum concentrations (mean +/- SD) of 4.7 +/- 0.3 micrograms/ml and 3.2 +/- 0.6 micrograms/ml, respectively. Twice this dose (30 mg of OTC/kg, every 8 hours) given IM caused anorexia and diarrhea.     

Isolation and Identification of Orf Virus from Jiangsu Province and Phylogenetic Analysis of Their B2L Gene

The study was aimed to investigate prevalence of Orf virus (ORFV) in Jiangsu province in recent years and control Orf better. A total of 121 tissue samples were collected in some farms from 2013 to 2015 and subjected to PCR detection, viral isolation and phylogenetic analysis of B2L gene. Four samples were ORFV positive by PCR. The viruses were isolated by passaging in ovine fetal turbinate (OFTu) cells and MDBK cells, and were named as ORFV/Ovis/XZ/Jiangsu/2015/China, ORFV1/Ovis/DT/Jiangsu/2015/China, ORFV2/Ovis/DT/Jiangsu/2015/China and ORFV/Ovis/SL/Jiangsu/2015/China,respectively. The B2L gene was amplified and sequenced for the phylogenetic study. The nucleotide homology of these 4 strains was 98.5% to 100.0%. ORFV/Ovis/XZ/Jiangsu/2015/China, ORFV1/Ovis/DT/Jiangsu/2015/China and ORFV2/Ovis/DT/Jiangsu/2015/China, gathered into a cluster with SC-JY, GX-YB, JS-FX isolates and the nucleic acid homology of these strains was 97.8% to 100%. ORFV/Ovis/SL/Jiangsu/2015/China gathered into a cluster with LiaoNing, HuB and Gansu isolates, the nucleic acid homology was 98.8% to 98.9%. The nucleic acid homologies of 4 strains and ORFV strain China vaccine was 96.8% to 98.1%. The result showed that the ORFVs in Jiangsu province might be from different source. For controlling the spreading of this virus, it was necessary to carry out deep epidemiological survey in Jiangsu province.     

心叶驼绒藜胚器官培养及再生体系的研究

以心叶驼绒藜无菌苗叶片、茎段、幼根为外植体,对通过器官发生的途径发育成再生植株进行了研究。结果表明：（1）、无菌苗的建立：用1/2MS＋琼脂＋蔗糖,出苗率高（90%以上）、苗壮。（2）、诱导培养基：用MS1＋IAA0.2mg/L＋KT0.1mg/L培养基,出愈率低,没有获得再生植株;用MS2＋IAA0.2mg/L＋KT0.2mg/L、MS3＋IAA0.1mg/L＋KT0.1mg/L培养基,叶片、茎段具有再生植株发育的能力,出愈率较高。（3）、生根培养基：用1/2MS＋NAA0.1 mg/L,生根率高、周期短;用1/2MS＋IAA0.1 mg/L,生根率低,周期长。     

甘肃高山细毛羊妊娠母羊营养需要量估测

借助其他品种绵羊营养需要的一些研究方法,结合甘肃高山细毛羊四季牧草干物质采食量、营养物质摄入量、采食牧草营养成分、牧草营养物质消化率以及生产相关数据,采用析因法估测甘肃高山细毛羊能量和蛋白质需要量:成年母羊维持代谢能需要量为6.58 MJ/d(每kg代谢体重维持代谢能需要为0.417 MJ/W^0.75),维持蛋白质需要量107.62 g/d;妊娠前期代谢能需要量为12.54 MJ/d,粗蛋白质需要量125.19 g/d;妊娠后期代谢能需要量为12.86 MJ/d,粗蛋白质需要量为146 g/d;通过对比研究表明,甘肃高山细毛羊成年母羊钙需要量约为8.22~9.61 g/d,磷需要量为4.98~5.87 g/d,食盐需要量为9~16 g/d。     

红茂草中精油成分分析及抑菌活性的研究

为红茂草药物的进一步开发和利用,本试验以甘肃平凉地区野生药用植物红茂草为研究对象,采用水蒸气提取、蒸馏萃取（SDE）提取、超临界CO₂萃取等3种方法,分别提取红茂草中的精油成分;以GC-MS对其精油进行化学成分分析,并将超临界CO₂萃取的红茂草精油采用平板菌落计数法对金黄色葡萄球菌、大肠杆菌和绿脓杆菌进行抑菌活性测定。结果显示,三种提取方法中,水蒸气提取法和蒸馏萃取提取法所得精油为橙黄色,有浑浊,超临界CO₂萃取法所得精油为淡黄色,澄清液体。3种方法所得精油均含有24种共同成分,以超临界CO₂萃取法最优,其提取率为2.1220%,水蒸气提取法最低,提取率为1.7870%;将超临界CO₂萃取的红茂草精油作用于3种供试菌,测定其最小抑菌浓度（MIC）分别为：金黄色葡萄球菌0.2500 g/mL、大肠杆菌0.3000 g/mL,绿脓杆菌0.2600 g/mL,抑菌效果显著。研究结果表明,超临界CO₂萃取是提取红茂草精油的最佳方法,通过超临界CO₂萃取法提取的红茂草精油对金黄色葡萄球菌、大肠杆菌和绿脓杆菌具有良好的抑菌效果。     

山羊碱性氨基酸转运载体基因SLC3A1cDNA序列克隆及表达

试验旨在克隆山羊碱性氨基酸转运载体基因SLC3A1 cDNA序列,探究其表达的组织特异性及其在小肠中的表达发育模式。参考GenBank已发表的绵羊、牛SLC3A1基因mRNA序列设计引物,克隆山羊SLC3A1基因的cDNA序列,采用实时荧光定量PCR的方法分析其在1日龄山羊11种组织及其在1日龄、6月龄、8月龄、10月龄、12月龄山羊十二指肠、空肠、回肠中的mRNA表达情况。结果表明,成功克隆得到了山羊SLC3A1基因cDNA序列,其与绵羊、牛、野猪、人、小鼠、大鼠的同源性分别为99%、97%、88%、86%、80%和79%。SLC3A1基因转录表达有明显的组织特异性,其在1日龄山羊肾脏、回肠、空肠、结肠中表达量依次降低(P<0.05),其他组织中表达量很低。同一月龄山羊,SLC3A1基因在不同肠段的表达量数值上回肠>空肠>十二指肠。SLC3A1基因在不同月龄山羊十二指肠表达量以1日龄山羊为最高(P<0.05),6~12月龄山羊相同肠段之间表达量无显著差异(P>0.05);空肠表达量随山羊年龄的增加均呈现逐步降低的趋势;回肠表达量在各个月龄山羊之间差异不显著(P>0.05)。结果说明,山羊碱性氨基酸转运载体基因SLC3A1的主要表达部位在肾脏和肠道,推测b^0,+碱性氨基酸转运系统转运氨基酸的主要部位为肾脏和肠道。SLC3A1基因在山羊小肠受肠段和发育阶段的影响,具有不同的表达发育模式。     

Preliminary characterisation of paramyxovirus isolated from a parrot.

A virus, designated 0121, which was isolated from a parrot, was shown to be a paramyxovirus, which was serologically related to the paramyxoviruses Bangor/flinch/N. Ireland/73 (Bangor) and Yucaipa/chicken/California/60(PMY). However, the 0121 virus differed in several properties from both the PMY and the Bangor viruses. The virus was not pathogenic for chickens. I was designated Paramyxovirus: Parrot/England/0121/74.     

猪轮状病毒VP4基因重组腺病毒的构建及抗体水平评价

【目的】构建猪轮状病毒（Porcine rotavirus,PoRV）流行株PoRV G9P[23]型的VP4基因重组腺病毒,为开发PoRV候选基因工程疫苗奠定基础。【方法】参考GenBank中流行株PoRV G9P[23]型的VP4基因序列（登录号：MH898990.1）合成PoRV VP4基因,将得到的目的基因与腺病毒穿梭载体pAdTrack-CMV进行重组,转化大肠杆菌Top10感受态细胞,构建腺病毒穿梭载体pAdTrack-CMV-VP4;腺病毒穿梭载体经过Pme Ⅰ内切酶线性化处理后与含有腺病毒骨架pAdEasy-1的大肠杆菌BJ5183感受态细胞进行同源重组获得重组质粒pAd-VP4,对重组质粒进行Pac Ⅰ酶切鉴定,并转化大肠杆菌DH5α感受态细胞。将重组质粒转染HEK293A细胞获得重组腺病毒rAd-VP4,对该重组腺病毒进行扩大培养并测定重组腺病毒的半数组织培养感染剂量（TCID₅₀）;通过RT-PCR检测其体外表达情况,Western blotting检测其反应原性;将制备的重组腺病毒用不同病毒滴度和不同免疫次数对小鼠进行腹腔免疫,收集血清通过ELISA法测定IgG抗体水平。【结果】RT-PCR扩增出1条大小为2 343 bp的rAd-VP4重组腺病毒条带,测序结果正确,表明重组腺病毒rAd-VP4构建成功,测得rAd-VP4病毒滴度为10^6.5 TCID₅₀,Western blotting结果表明,重组腺病毒rAd-VP4在蛋白水平上得到了正确表达,蛋白的分子质量约为87 ku。小鼠IgG抗体检测结果表明,在用10^6.5 TCID₅₀ rAd-VP4免疫后的第35和42天,小鼠血清中的IgG抗体水平显著高于10^5.3 TCID₅₀ TGE-PED-PRV三联活疫苗IgG抗体水平（P<0.05）;10^6.5 TCID₅₀ rAd-VP4在免疫后第35和42天产生的抗体水平显著高于10^5.5和10^4.5 TCID₅₀ rAd-VP4（P<0.05）,而10^5.5 TCID₅₀ rAd-VP4在免疫后第28天产生的抗体水平显著高于10^6.5和10^4.5 TCID₅₀ rAd-VP4（P<0.05）。10^6.5 TCID₅₀ rAd-VP4的2次免疫和3次免疫产生的IgG抗体在不同免疫时间均差异不显著（P>0.05）。【结论】本研究成功构建了重组腺病毒rAd-VP4,其病毒滴度为10^6.5 TCID₅₀。10^6.5和10^5.5 TCID₅₀ rAd-VP4分别在第42和28天产生较高的IgG抗体水平,2次免疫和3次免疫对产生IgG抗体水平均无显著影响。试验结果可为开发PoRV重组腺病毒候选疫苗提供参考。     

马流感病毒A/马/青海1/94株亚型鉴定及其HA基因序列特征

本试验用AI标准阳性分型血清对我国马流感病毒A／马/青海/1／94进行了血凝素(H)和神经氨酸酶(N)的鉴定，并与马流感病毒吉林株、黑龙江株、北京株及国际标准株A／切ukx／Mgxd／2／63进行了对比，结果显示，1944年在我国青海省暴发的马流感病毒的亚型为H3N8；以反转录。聚合酶链式反应(RT-PCR)扩增该株的血凝素基因，并克隆到pGEM-Teasy载体上，采用双脱氧末端终止法测定该cDNA片段共1738个核苷酸序列，并推导出其编码的565个氨基酸的序列。利用Genbank blast和基因进化树分析软件分析各毒株之间的亲缘关系，发现A／马/青海/1/94与1992年香港马流感分离株存在较近的亲缘关系。     

猪链球菌病C、D、E三价灭活蜂胶疫苗的研制

以C、D、E群猪链球菌强毒株经培养后灭活、浓缩、混合,用蜂胶作佐剂(10mg/头剂),制成C、D、E三价蜂胶灭活疫苗,并在实验室内进行安全性试验及免疫效果实验。结果表明,该疫苗对实验猪、小鼠及家兔均安全;小鼠免疫21天后攻毒,对C55138株的保率护分别是4/5、5/5、4/5;C55938株的保护率分别是3/5、4/5、4/5;C55914株的保护率分别是4/5、3/5、3/5,对照组均5/5死亡;猪免疫21天后以C55138株攻毒,0401试制疫苗保护率为3/4外,其余均4/4保护;对C55938株攻毒保护率三批分别为4/4、2/4、3/4。对照猪均4/4死亡,疫苗免疫保护效果良好。     

Metronidazole: a method for its determination in biological fluids and its disposition kinetics in the dog

A method for the analytical determination of metronidazole concentrations in biological tissues was developed using high performance liquid chromatography. The procedure was employed to investigate the pharmacokinetics of metronidazole in dogs following intravenous and oral administration (44 mg/kg). The overall elimination rate constant β was 0.0027 ± 0.0005 min^-1, the apparent specific volume of distribution (V'_d) was 0.948 ± 0.096 L/kg overall clearance (Cl_B) was 2.49 ± 0.54 ml/kg/min and the rate constant for absorption K_ab was 0.0456 ± 0.0353 min^-1. Oral bioavailability was high but variable (59%–100%). Implications of these data for chemotherapy of infections caused by anaerobic bacteria, trichomonads, and Giardia and for the sensitization of hypoxic neoplastic cells to radiotherapy are discussed.     

Pharmacokinetic study of dipyrone metabolite 4-MAA in the horse and possible implications for doping control

The pharmacokinetic behaviour of dipyrone metabolite 4-MAA in serum was determined in seven horses of different breeds after a single intravenous dose administration. A biexponential formula was fitted to the serum concentration vs. time data. The median half-life of the elimination phase ( t _1/2β) was 4.85 h (range 5.04 h), the median volume of distribution ( V d_area) was 1.85 L/kg (range 3.2 L/kg) and median of total clearance was 4.0 mL/min/kg (range 2.3 mL/min/kg).     

干扰Smad3促进山羊脂肪细胞分化

旨在克隆山羊Smad3的基础上，明确其组织和细胞表达谱，最终阐明干扰Smad3基因对山羊肌内和皮下脂肪细胞分化的影响。本研究选用5只体况良好的1周龄简州大耳羊，空腹24 h后屠宰并采集相应组织和细胞进行试验。利用RT-PCR技术克隆山羊Smad3基因cDNA区序列并进行生物信息学分析，利用实时荧光定量PCR（real-time quantitative PCR，qPCR）技术检测Smad3基因的组织和细胞时序表达水平；并且合成靶向Smad3的siRNA，采用油红O染色从形态学上明确干扰Smad3对山羊前体脂肪细胞成脂分化的影响，利用qPCR检测干扰Smad3对脂肪细胞分化标志基因C/EBPα、C/EBPβ、LPL、SREBP1、AP2、PPARγ、Pref-1、KLF3、KLF4、KLF6、KLF7、KLF8、KLF9、KLF10和KLF15以及Smads相关基因Smad2、Smad4、Smad7和TGF-β1基因mRNA表达水平的影响，探讨可能的作用机制。结果，获得山羊Smad3基因1 449 bp，其中CDS区序列为1 278 bp，编码425个氨基酸；Smad3在山羊各组织中具有广泛表达特性，且在肾脏中的表达水平最高（P<0.01）；Smad3均在山羊肌内和皮下两种脂肪细胞诱导分化的36 h表达量最低，极显著低于在未分化前体脂肪细胞中的表达丰度（P<0.01）；干扰Smad3后发现显著促进了山羊肌内和皮下脂肪细胞中脂滴的聚集，且脂肪细胞分化标志基因、KLF3、KLF4、KLF8、KLF9、KLF10和KLF15的表达水平显著上升（P<0.05），Pref-1的相对表达水平极显著下降（P<0.01），同时干扰Smad3基因下调了Smad2、Smad4和Smad7基因的相对表达水平（P<0.05）。研究结果指出，干扰Smad3促进山羊脂肪细胞分化，且可能通过调控脂肪细胞分化标志基因C/EBPα、C/EBPβ、LPL、SREBP1、AP2、Pref-1、KLF3、KLF4、KLF8、KLF9、KLF10和KLF15等及协同Smad2、Smad4和Smad7的表达来实现的。     

雌激素调控奶牛乳腺上皮细胞凋亡及生长周期作用机制的研究

试验旨在研究雌激素对奶牛乳腺上皮细胞（BMECs）凋亡及生长周期的影响。通过添加MAPK/ERK信号通路阻断剂探索雌激素调控BMECs凋亡及生长周期具体的作用机制,采用流式细胞仪检测细胞凋亡及周期的变化情况,实时荧光定量PCR检测Bcl-2、Caspase3及CyclinD1基因mRNA的表达丰度。结果显示,对照组BMECs凋亡率极显著低于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01）,Bcl-2 mRNA表达丰度极显著高于BMECs+PD98059组（P<0.01）,Caspase3 mRNA表达丰度显著低于BMECs+PD98059组（P<0.05）;对照组细胞比例在G1期显著高于BMECs+E₂组（P<0.05）,极显著低于BMECs+E₂+PD98059组（P<0.01）,S期细胞比例极显著高于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01）,G2期细胞比例极显著低于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01）;对照组CyclinD1 mRNA的表达丰度极显著高于BMECs+PD98059组（P<0.01）;BMECs+E₂+PD98059组的Bcl-2 mRNA的表达量极显著高于BMECs+PD98059组（P<0.01）,Caspase3 mRNA的表达量显著低于BMECs+PD98059组（P<0.05）。结果表明,MAPK/ERK信号通路参与BMECs的增殖及细胞生长周期调节的过程,且雌激素可通过MAPK/ERK信号通路抑制BMECs的凋亡,MAPK/ERK信号通路可能参与由雌激素调控的细胞生长周期的进程。     

江苏省羊口疮病毒的分离鉴定及B2L基因的遗传进化分析

为了解江苏省近年来羊口疮病毒（Orf virus,ORFV）的流行情况,更好地控制江苏地区的羊口疮病,2013~2015年采集江苏部分地区羊口疮疑似病料121份,进行病毒分离鉴定及其B2L基因的遗传进化分析。结果显示,样品中ORFV PCR检测阳性4份,用胎羊鼻甲骨细胞（OFTu）和MDBK细胞进行病毒分离,分离到4株病毒。这4株病毒分别命名为ORFV/Ovis/XZ/Jiangsu/2015/China、ORFV1/Ovis/DT/Jiangsu/2015/China、ORFV2/Ovis/DT/Jiangsu/2015/China和ORFV/Ovis/SL/Jiangsu/2015/China。扩增ORFV的B2L基因全长并绘制遗传进化树。B2L基因序列分析显示,4株分离株之间的核酸同源性为98.5%~100.0%。遗传进化树显示,ORFV/Ovis/XZ/Jiangsu/2015/China株、ORFV1/Ovis/DT/Jiangsu/2015/China株和ORFV2/Ovis/DT/Jiangsu/2015/China株与SC-JY、GX-YB、JS-FX株聚成一簇,核苷酸同源性为97.8%~100.0%。ORFV/Ovis/SL/Jiangsu/2015/China与LiaoNing、HuB、Gansu株亲缘关系接近,核苷酸同源性为98.8%~98.9%。4株分离株与中国疫苗株的核苷酸同源性为96.8%~98.1%。结果表明,江苏地区的ORFV来源可能不同,有必要开展更为深入的流行病学调查,为防控江苏地区的羊口疮奠定基础。     

盐胁迫对裸果木幼苗光合特性的影响

本研究以一年生裸果木幼苗为材料，采用盆栽育苗方式，设计7个不同浓度NaCl溶液（CK、0.2%、0.4%、0.6%、0.8%、1.0% 和1.2%）模拟盐分胁迫，测定分析了其叶片气体交换参数、光响应曲线及叶绿素荧光参数的变化规律。结果表明：随着盐浓度的增加，叶片净光合速率（P_n）、气孔导度（G_s）和蒸腾速率（T_r）均呈下降趋势，NaCl浓度≥0.4%时，各处理P_n、G_s和T_r均显著低于CK；气孔限制值（L_s）和瞬时水分利用效率（WUE）随盐浓度增加呈先升后降趋势，胞间CO₂浓度（C_i）呈相反趋势，NaCl浓度为0.4%时，L_s达到最大值，而C_i达到最小值，说明P_n下降以气孔限制因素为主，而当NaCl浓度≥0.6%时，以非气孔限制为主要因素。随着盐胁迫程度的增大，最大净光合速率（P_nmax）、暗呼吸速率（R_d）、光饱和点（L_SP）、表观量子效率（A_QY）逐渐降低，光补偿点（L_CP）逐渐增加，表明盐分抑制了幼苗对光的吸收、利用和转换能力。叶片PSII潜在活性（F_v/F₀）、原初光能转化效率（F_v/F_m）、实际光化学效率（Φ_PSII）、电子传递速率（ETR）和光化学猝灭系数（q_p）随着盐浓度的增加呈下降趋势；非光化学猝灭系数（NPQ）在≤0.4%NaCl处理下较CK显著增加，盐浓度≥0.8%时，NPQ显著下降。基质的NaCl浓度在0.2%和0.4%时，裸果木叶片P_n、F_v/F₀、F_v/F_m下降不显著，WUE有所提高，PSII系统可以通过耗散过剩的光能保护光合机构，表现出一定的耐盐性；但盐浓度超过0.6%时，光合生态幅变窄，光合机构受到明显破坏，显著抑制了光合作用能力。