似鲶高原鳅温和气单胞菌的分离鉴定及其感染的病理损伤

2013年9月,四川雅安某似鲶高原鳅养殖场发生以体表出血、皮肤溃疡、内脏器官肿大、出血与坏死为特点的疾病。从自然发病的似鲶高原鳅肝与肾分离到1株G-短杆菌(XJJ130928),其在BHI平板上28℃培养24 h,形成灰白色、表面光滑、边缘整齐、半透明状微隆的圆形菌落,人工感染证实其为本次似鲶高原鳅发病的病原菌。根据分离菌株的形态学和生理生化检测结果初步判定其为温和气单胞菌Aeromonas sobria;进一步的16S rRNA和gyrB基因序列分析表明,分离株的16S rRNA与gyrB基因序列(GenBank登录号:KF761305与KJ139988)在GenBank中进行Blast比对与A.sobria同源性最高。在以分离株16S rRNA和gyrB基因序列及GenBank中同源性较高的序列构建的系统发育树上,分离株与A.sobria聚为一族,其同源性分别为96.0%~99.0%与94.5%~98.0%,结合生理生化特性鉴定分离菌为A.sobria。该菌对头孢西丁、亚胺培南、强力霉素、氟苯尼考和氧氟沙星等敏感,对磺胺甲基异恶唑、洛美沙星、环丙沙星中度敏感,对氨苄青霉素、链霉素、卡那霉素耐药。组织病理学观察发现,A.sobria感染似鲶高原鳅对多组织器官都造成明显的病理损伤,尤其是肝、肾、脾、肠的损伤较为严重,表现为明显的淤血、出血,变性,坏死及炎症细胞浸润。     

红鳍东方鲀病原鱼肠道弧菌的生物学特性研究

对引起红鳍东方鲀发病死亡的病原细菌进行了分离和主要生物学特性研究,包括病原性、形态特征、理化特性、16S rRNA基因序列及其系统发育学分析、胞外酶及溶血素活性、K抗原及耐药性等.结果表明,引起红鳍东方纯发病死亡的病原细菌为弧菌属(-Vibrio Pacini 1854)的鱼肠道弧菌(V.ichthyoenteri Ishimaru,et al.1996),2株代表菌株16S rRNA基凶序列(GenBank登录号分别为:EF611424和EF635304)与GenBank数据库中鱼肠道弧菌的同源性在98%-100%,且在构建的MP系统发生树中与鱼肠道弧菌聚为一个分支.分离菌不具有淀粉酶、蛋白酶、脂肪酶、DNA酶、脲酶、明胶酶和卵磷脂酶活性,且在含7%家兔脱纤血液营养琼脂培养基上不溶血.不具有K抗原.人工感染试验中分离菌对红鳍东方纯表现出明显的致病性.药敏试验结果显示,4株分离菌对供试37种抗菌药物中的苯唑青霉素和杆菌肽2种耐药.     

突发疫情中炭疽芽孢杆菌的分离及鉴定

对疑似炭疽感染病牛牛肉标本和牛血污染土壤标本进行了病原菌分离,经菌落形态和菌体形态观察、血清学实验和生化鉴定,证明分离到的细菌为炭疽芽孢杆菌。为进一步了解其特性,分别用保护性抗原、水肿因子和荚膜基因特异性引物对2株菌进行PCR扩增。结果显示,这两株菌有两个毒力相关质粒pX01和pX02,为有毒株。序列测定表明,这两株菌基因间同源性达99%,这两株菌与GenBank中炭疽芽孢杆菌A2012株、Ames Ancestor株和A16R疫苗株同源性达99%。     

兔出血症病毒（RHDV）WHNRH株的分离鉴定及基因组全序列的测定与分析

从发病长毛兔中分离鉴定了兔病毒性出血症病毒WHNRH株。参考GenBank中已登录的RHDV毒株序列对RHDV WHNRH分离株进行了全基因组序列测定与分析。设计5对扩增区段相互重叠的RHDV特异性引物,扩增除5′和3′末端以外的序列,采用设计锚引物的5′RACE方法以及针对RHDV 3′末端的polyA结构设计引物获得了RHDV WHNRH株的5′和3′末端序列。胶回收各PCR产物,连接pMD 18-T克隆载体,测得RHDV WHNRH分离株的基因组全长为7437nt(不包括polyA),与GenBank公布的全部共6株RHDV全基因序列进行同源性比较分析,同源性在89.0%~97.1%之间,ORF1同源性为89.0%~97.1%,编码氨基酸序列的同源性为95.2%~98.7%;ORF2的核酸苷序列同源性为92.1%~97.7%,编码氨基酸序列的同源性为94.1%~96.6%。     

小熊猫犬瘟热病及病原研究

1999年2月和1999年7月,重庆动物园和雅安碧峰峡生态动物园喂养的小熊猫分别大面积爆发和流行犬瘟热病,重庆动物园小熊猫死亡11只,死亡率达100％,碧峰峡动物园小熊猫死亡4只,死亡率为25％,两地发病猫所表现的临床症状和病理解剖变化不完全一致,但均有血便、呼吸急促、不食、精神差、肺出血充血等类似症状。从碧峰峡动物园患病小熊猫分离出的犬瘟热毒株（cdv2株）对VEFO细胞适应性比重庆动物园分离的毒株(cdv1株）强,其TCID50达10^－6。两地分离的毒株均能同抗犬瘟热阳性高免血清反应。此外,用抗犬瘟热高免血清治疗,成功地挽救了12只病重的小熊猫。     

圈养小熊猫几种病毒的PCR检测

本文采用巢式PCR/ RT-PCR 方法,对我国10 个动物园中无临床症状的圈养小熊猫的71 个肛拭子和61 个唾液拭子样品,进行犬瘟热病毒(CDV)、犬细小病毒(CPV)、犬冠状病毒(CCV)、犬腺病毒(CAV)和犬疱疹病毒(CHV)的检测,以评估我国圈养小熊猫是否面临这几种病毒的威胁。对阳性PCR 结果进行测序分析,并与GenBank 上的序列进行比较。结果,在肛拭子样品中检测到3 个CPV 和6 个CCV 阳性结果,经测序后,与GenBank 上序列的同源性分别达99% 和100% 。而在唾液拭子样品中没有检测到任何阳性结果,且CDV、CAV和CCV 的检测结果均为阴性。从阳性CPV 的肛拭子样品中分离到一株细小病毒毒株,表明圈养小熊猫已受到细小病毒和犬冠状病毒的感染,今后应加强这两种病毒的预防工作。本文所采用的PCR 方法检测病毒性疾病,能检测到微量的病毒模板,可对小熊猫病毒性感染进行早期诊断。     

紫色色杆菌感染临床特症及耐药谱分析

目的 研究紫色色杆菌感染的临床特征与病原体的生物学特性以及耐药分析,以提高临床对紫色色杆菌感染的诊断和治疗.方法 将患者的血液、浓液标本进行培养,分离并把分离获得的菌株在VITEK-32微生物鉴定仪中进行种的鉴定及药物敏感性试验,用小白鼠进行该病原体的毒力试验.结果 12种抗菌药物中舒普深、亚胺培南、特治星（派拉西林/他唑巴坦）、马斯平（头孢吡肟）及喹喏酮类抗菌药物对紫色色杆菌均有较强的抗菌作用,而氨苄西林、头孢唑啉、庆大霉素、妥布霉素、氨曲南、头孢曲松和头孢他啶等抗菌药物抗菌效果不佳.毒力试验结果表明该菌有较强毒力.结论 紫色色杆菌是一种革兰阴性杆菌,毒力强.可引起感染部位脓肿,并可侵入体内各脏器及血液引发脏器的脓肿,脓毒血症是该菌感染的临床特症,重者可引起各脏器功能衰退,中毒性休克甚至死亡.紫色色杆菌感染病情急,进展快,死亡率高,因此早发现,及早有效性治疗极为重要.     

Viili乳制品中干酪乳杆菌的分离鉴定

从引进Viili乳制品中筛选、鉴定出3种优良乳酸菌。采用平板分离法从Viili乳制品中分离3株乳酸菌,通过表型1、6S rRNA的PCR扩增、克隆、测序鉴定。3株乳酸菌的表型鉴定结果符合伯杰细菌鉴定手册中乳杆菌属的干酪乳杆菌,16S rRNA序列同源性分析结果表明3株分离菌与干酪乳杆菌的同源性分别为99.93%、100.00%9、9.78%,从Viili乳制品分离到3株干酪乳杆菌。     

慢性蜜蜂麻痹病毒Ch1株的编码区测序及其序列的分析

对中国分离株慢性蜜蜂麻痹病毒(Chronic bee paralysis virus,CBPV)Ch1编码区全基因序列进行克隆、测序、分析。利用RT-PCR方法和生物信息学软件,对本实验室分离到的Ch1株CBPV编码区的基因序列进行克隆,测序,与GenBank收录的CBPV毒株进行同源性比较,并以RdRp为靶基因构建了遗传进化树。结果显示,CBPV Ch1株的编码区由RNA1(GenBank No.KU950353)和RNA2(GenBank No.KU950354)两部分构成,全长5 979个核苷酸。其中RNA1片段全长3 674个核苷酸,编码3个开放阅读框,RNA2片段全长2 305个核苷酸,编码4个开放阅读框,RNA2片段中ORF2和ORF3,可能编码两个结构蛋白,分别命名为SP1和SP2。RNA1和RNA2核苷酸序列与2005年法国分离株Fr2核苷酸序列同源性最高,分别为96.1%和95.5%,但预测蛋白SP1核苷酸序列同源性与2006年乌拉圭分离株Ur1核苷酸序列同源性最高(96.9%)。基于RdRp为靶基因进行了遗传进化分析表明,Ch1株与Fr2株位于同一分支,且在该区域,Ch1株与Fr2株的核苷酸序列同源性最高(96.5%)。本实验成功分离到一株CBPV(KU950353,KU950354),并命名为Ch1株,完成了Ch1株CBPV的编码区的序列测定以及核苷酸序列与推导的氨基酸序列同源性比较及遗传进化分析,为研究CBPV的致病机制和免疫机制提供重要信息。     

禽流感病毒分离株NS基因同源性及等位基因类型分析

目的　克隆测定国内具有代表性的禽流感病毒 (AIV)的非结构 (NS)蛋白基因核苷酸序列 ,分析其同源性和等位基因类型 ,为进一步探索禽流感NS蛋白抗体监测方法奠定基础。方法　经RT PCR扩增了国内 3株H9N2、2株H5N1、2株H7N2亚型AIV分离株的NS蛋白基因 ,并把扩增的基因片段克隆到pGEM T载体中测序 ,将测序结果与GenBank中的核苷酸序列进行同源性比较 ,绘制基因进化树。结果　经测序获得了各AIV分离株NS基因的完整编码序列。同源性分析表明 ,3株H9亚型AIV的NS基因之间的同源性为 96 %～ 98% ;两株H5亚型AIVNS基因同源性为 91 6 % ;两株H7亚型AIV的NS基因同源性为 98 9%。H5和H9亚型分离株的NS基因之间的同源性均高于 90 % ;而H7N2亚型分离株与其它两种亚型分离株的NS基因同源性约为 6 0 %～ 70 %。在AIVNS基因系统发育进化树中 ,H5、H9亚型分离株都处于等位基因A群内 ;3株H9亚型分离株的进化关系较近 ,与香港、广东的部分H5N1病毒株起源相同 ,而 2株H5病毒的NS基因则处于不同分枝内 ;2株H7亚型分离株的NS基因都处于等位基因B群内 ,进化关系较近。结论　这 7株国内AIV分离株的NS基因之间的同源性差异较大 ,约为 6 0 %～ 99% ,且包括A、B两种类型的等位基因     

Negative phototropic response of rhizoid cells in the fern Adiantum capillus-veneris

In general, phototropic responses in land plants are induced by blue light and mediated by blue light receptor phototropins. In many cryptogam plants including the fern Adiantum capillus-veneris, however, red as well as blue light effectively induces a positive phototropic response in protonemal cells. In A. capillus-veneris, the red light effect on the tropistic response is mediated by phytochrome 3 (phy3), a chimeric photoreceptor of phytochrome and full-length phototropin. Here, we report red and blue light-induced negative phototropism in A. capillus-veneris rhizoid cells. Mutants deficient for phy3 lacked red light-induced negative phototropism, indicating that under red light, phy3 mediates negative phototropism in rhizoid cells, contrasting with its role in regulating positive phototropism in protonemal cells. Mutants for phy3 were also partially deficient in rhizoid blue light-induced negative phototropism, suggesting that phy3, in conjunction with phototropins, redundantly mediates the blue light response.     

Interaction of phloretin with the anion transport protein of the red blood cell membrane

Phloretin is an inhibitor of anion exchange and glucose and urea transport in human red cells. Equilibrium binding and kinetic studies indicate that phloretin binds to band 3, a major integral protein of the red cell membrane. Equilibrium phloretin binding has been found to be competitive with the binding of the anion transport inhibitor, 4,4′-dibenzamido-2,2′-disulfonic stilbene (DBDS), which binds specifically to band 3. The apparent binding (dissociation) constant of phloretin to red cell ghost band 3 in 28.5 mM citrate buffer, pH 7.4, 25°C, determined from equilibrium binding competition, is $\text{1.8 ± 0.1}\text{μM}$. Stopped-flow kinetic studies show that phloretin decreases the rate of DBDS binding to band 3 in a purely competitive manner, with an apparent phloretin inhibition constant of $\text{1.6 ± 0.4}\text{μM}$. The pH dependence of equilibrium binding studies show that it is the charged, anionic form of phloretin that competes with DBDS binding, with an apparent phloretin inhibition constant of 1.4 μM. The phloretin binding and inhibition constants determined by equilibrium binding, kinetic and pH studies are all similar to the inhibition constant of phloretin for anion exchange. These studies suggest that phloretin inhibits anion exchange in red cells by a specific interaction between phloretin and band 3.     

Modes of control by the circadian oscillator and the hourglass mechanism of the activities of cytoplasmic and chloroplast glyceraldehyde 3-phosphate dehydrogenases in Lemna gibba G3

Two types of clocks, i.e., the circadian oscillator and thehourglass mechanism, which under continuous light and darknessrespectively control the mutually inverse temporal changes inthe activities of Cyt-NAD-GPD and Chl-NADP-GPD of Lemna gibbaG3, were studied. Both clocks controlled the apparent Km values,not the Vmax values, of the GPD reactions for their substrateand coenzymes. A red light pulse inserted 3 hr after the onsetof the dark period eliminated the sigmoidal changes in darkness,but evoked rhythmical changes which otherwise did not occurin continuous darkness. Thus, the photosynthetic rhythm, ifpresent, would not sustain the GPD rhythms. This effect of ared light pulse was not nullified by a subsequent far red lightpulse. A far red light pulse given at the 3rd hour of an extendeddark period made conspicuous the sigmoidal changes in activityof GPDs in the dark period, and its effect was nullified bya subsequent red light pulse, suggesting that phytochrome isinvolved in the hourglass mechanism. (Received September 26, 1978; )     

Ultraviolet A-specific induction of anthocyanin biosynthesis in the swollen hypocotyls of turnip (Brassica rapa)

Ultraviolet A (UV-A)-mediated regulation of anthocyanin biosynthesis was investigated in swollen hypocotyls of the red turnip 'Tsuda'. The shaded swollen hypocotyls which contained negligible anthocyanin were exposed to artificial light sources including low fluence UV-B, UV-A, blue, red, far-red, red plus UV-A, far-red plus UV-A, and blue plus red. Among these lights, only UV-A induced anthocyanin biosynthesis and co-irradiation of red or far-red with UV-A did not affect the extent of UV-A-induced anthocyanin accumulation. The expression of phenylalanine ammonia lyase (PAL; EC 4.3.1.5), chalcone synthase (CHS; EC 2.3.1.74), flavanone 3-hydroxylase (F3H; EC 1.14.11.9), dihydroflavonol 4-reductase (DFR; EC 1.1.1.219), and anthocyanidin synthase (ANS; EC 1.14.11.19) genes was increased with time during a 24 h exposure to UV-A. In contrast, irradiation with red, blue, UV-B, and a combination of blue with red failed to induce CHS expression. Microarray analysis showed that only a few genes, including CHS and F3H, were induced significantly by UV-A, while a separate set of many genes was induced by low fluence UV-B. The UV-A-specific induction of anthocyanin biosynthesis and the unique gene expression profile upon UV-A irradiation as compared with blue and UV-B demonstrated that the observed induction of anthocyanin biosynthesis in red turnips was mediated by a distinct UV-A-specific photoreceptor, but not by phytochromes, UV-A/blue photoreceptors, or UV-B photoreceptors.     

Kinetics and time dependence of the effect of far red light on the photoperiodic induction of flowering in wintex barley

Flowering in the long day plant Hordeum vulgare L. var. Wintex barley was enhanced by the addition of far red light to the main light portion of the photoperiod. Far red energy was provided to produce quantum flux ratios (660/730 nm) and phytochrome photoequilibria (Pfr/total phytochrome) equivalent to those reported both beneath a leaf canopy and outside a canopy at twilight. The photoperiodic requirement for long days can be completely eliminated by the addition of far red light. However, both the effect of extending the photoperiod without far red and the addition of far red to 12-hour photoperiods were suboptimal. Maximal stimulation was achieved only when far red was added to continuous light. The duration of the period of maximal apex elongation rate, as well as the reduction of the time required for floral initiation, were saturated by three inductive cycles. When far red energy was provided intermittently during 3 days of continuous light, the ability to respond varied in a circadian manner. This enhancement of flowering by far red appears to be mediated by the “high irradiance response” of phytochrome.     

Phytochrome and Seed Germination. I. Temperature Dependence and Relative P(FR) Levels in the Germination of Dark-germinating Tomato Seeds

Germination of the dark-germinating seeds of 3 varieties of tomato is controlled by the phytochrome system. Germination is inhibited by far red radiation and repromoted by red applied after far red. At low temperatures, 17 to 20°, a single, low energy far red irradiation is sufficient to inhibit germination in all 3 varieties. At higher temperatures far red is less effective in the inhibition of the germination of the tomato seeds. The phytochrome fraction present as P_FR in the dark-germinating seeds of the Ace variety is about 40% of the total phytochrome present.     

The plastid genome of the red alga Laurencia

We present the 174,935 nt long plastid genome of the red alga Laurencia sp. JFC0032. It is the third plastid genome characterized for the largest order of red algae (Ceramiales). The circular‐mapping plastid genome is small compared to most florideophyte red algae, and our comparisons show a trend toward smaller plastid genome sizes in the family Rhodomelaceae, independent from a similar trend in Cyanidiophyceae. The Laurencia genome is densely packed with 200 annotated protein‐coding genes (188 widely conserved, 3 open reading frames shared with other red algae and 9 hypothetical coding regions). It has 29 tRNAs, a single‐copy ribosomal RNA cistron, a tmRNA, and the RNase P RNA.     

Flowering of the long-day plant, Lemna gibba, under short-day schedules composed of red and far-red light

Flowering in Lemna gibba, a long-day duckweed, can be inducedunder a short-day condition when the photoperiodic regimes areR₇FR₃ (7 hr red followed by 3 hr far-red), R₅FR₅ and R₃FR₇.This indicates the necessity of a proper balance between redand far-red effects for flowering. The flowering induced bythese regimes is inhibited by a brief exposure to red givenat the start of darkness and this inhibition is reversed bysubsequent exposure to far-red. Thus, the red/far-red reversibleeffect is found only at the beginning of darkness for floweringof L. gibba. However, flowering of L. gibba is promoted by a red light breakgiven near the middle of a 14 hr dark period. The promotiveeffect is not reversed by subsequent exposure to far-red, i.e.,the effect of the red break converts from inhibition to promotionas when given later in the dark period, which suggests the involvementof a timing mechanism. (Received July 21, 1973; )     

Interaction between the effects of phytochrome and gibberellic acid on the senescence of Alstroemeria pelegrina leaves

Chlorophyll loss in the leaves of cut flowering branches of Alstroemeria pelegrina L. cv. Stajello, placed in water in darkness at 20°, was inhibited by irradiation with red light and by the inclusion of gibberellic acid (GA₃) in the water. The effects of red light were abolished when it was followed by far-red light. Effects of GA₃ and red light were additive over a range of GA₃ concentrations (0. 01–1 μ M ). Chlorophyll breakdown was increased by the inclusion of AMO-1618, ancymidol, or tetcyclasis in the water. The effect of these inhibitors of gibberellin synthesis was fully reversed by GA₃. The inhibition of chlorophyll breakdown by red light was absent when AMO-1618, ancymidol or tetcyclasis were included in the water. The results indicate that leaf yellowing is controlled by endogenous gibberellins and that the effect of phytochrome is mediated by gibberellin synthesis.