Increase in milt production by hormonal treatment in the pejerrey fish Odontesthes bonariensis (Valenciennes 1835)

In spite of interest in the cultivation of the pejerrey fish Odontesthes bonariensis (Cuvier & Valenciennes 1835), there are few studies on subjects required to advance this activity. One of the problems is the synchronization of female and male maturation to provide eggs and sperm for larval production. The low volume of expressible milt, either in wild or culture fish, is a major problem. The aim of this work was to study the effectiveness of the administration of different hormones on sperm production in pejerrey. Milt production was enhanced by the injection of human chorionic gonadotropin (hCG) (16.7‐fold increase, 625 IU kg^?1), carp pituitary extracts (13.5‐fold increase, 30 mg kg^?1), salmon pituitary extracts (12.8‐fold increase, 30 mg kg^?1), salmon‐type gonadotropin‐releasing hormone analogue (GnRH) (16.7‐fold increase, 10 μg kg^?1) and mammalian‐type GnRH analogue (10.8‐fold increase, 20 μg kg^?1). Sperm concentration, motility and the fertilization rate were not statistically different compared with control groups. It was also demonstrated that sperm could be obtained off‐season. Taken together, hCG is recommended to stimulate pejerrey spermiation because it is effective in low doses is inexpensive and is widely available.     

Midazolam,as a co‐induction agent,has propofol sparing effects but also decreases systolic blood pressure in healthy dogs

ObjectiveTo evaluate the effects of the co-administration of midazolam on the dose requirement for propofol anesthesia induction, heart rate (HR), systolic arterial pressure (SAP) and the incidence of excitement.Study designProspective, randomized, controlled and blinded clinical study, with owner consent.AnimalsSeventeen healthy, client owned dogs weighing 28 ± 18 kg and aged 4.9 ± 3.9 years old.MethodsDogs were sedated with acepromazine 0.025 mg kg^?1 and morphine 0.25 mg kg^?1 intramuscularly (IM), 30 minutes prior to induction of anesthesia. Patients were randomly allocated to receive midazolam (MP; 0.2 mg kg^?1) or sterile normal saline (CP; 0.04 mL kg^?1) intravenously (IV) over 15 seconds. Propofol was administered IV immediately following test drug and delivered at 3 mg kg^?1 minute^?1 until intubation was possible. Scoring of pre-induction sedation, ease of intubation, quality of induction, and presence or absence of excitement following co-induction agent, was recorded. HR, SAP and respiratory rate (f_R) were obtained immediately prior to, immediately following, and 5 minutes following induction of anesthesia.ResultsThere were no significant differences between groups with regard to weight, age, gender, or sedation. Excitement occurred in 5/9 dogs following midazolam administration, with none noted in the control group. The dose of propofol administered to the midazolam group was significantly less than in the control group. Differences in HR were not significant between groups. SAP was significantly lower in the midazolam group compared with baseline values 5 minutes after its administration. However, values remained clinically acceptable.Conclusions and clinical relevanceThe co-administration of midazolam with propofol decreased the total dose of propofol needed for induction of anesthesia in sedated healthy dogs, caused some excitement and a clinically unimportant decrease in SAP.     

Metallothionein induction in cultured fibroblasts and liver of a marine flatfish,the turbot,Scophthalmus maximus

Intraperitoneal injection of turbot with Cd induced the synthesis of a low molecular weight hepatic Cd-binding protein and a 500bp mRNA, which hybridised to a plaice metallothionein (MT) cRNA probe. The Cd-binding protein displayed cross-reactivity in a competitive ELISA with antiserum raised against rainbow trout MT and had the characteristic amino acid composition, metal stoichiometry and spectral characteristics of a Class I MT. Only one isoform was apparent on ion exchange chromatography. Southern blot analysis of DNA cleaved with four restriction enzymes suggested that only a single MT gene is present in turbot.In an established turbot fibroblast cell line, Cd induced MT mRNA and MT levels in a dose and time-dependent manner. MT was also induced by Cu, Hg and Zn but not Pb exposure. Physiological concentrations of glucocorticoids and sex hormones did not induce MT synthesis, although at high concentrations a positive response to corticosterone, dexamethasone, hydrocortisone or progesterone was observedin vitro indicating the possible presence of a functional steroid regulatory element in the fish MT gene.Abbreviations used AMP adenosine monophosphate - CHSE chinook salmon embryo - DMSO dimethyl sulphoxide - HPLC high performance liquid chromatography - KB kenacid blue - MT metallothionein - Mr molecular weight (kDaltons) - NR neutral red - PEG polyethylene glycol - RTH rainbow trout hepatoma - SDS sodium dodecyl sulphate - SSC 0.15M NaCl, 0.015M sodium citrate - TF turbot fibroblast     

人工诱导池塘养殖鳗鲡成熟产卵以及胚胎和仔鱼发育

降海鳗鲡一直是作为研究鳗鲡人工繁殖的材料,由于降海鳗鲡的资源衰退,选用池塘养殖鳗鲡作为亲本的研究成为热点。尽管人工诱导野生降海鳗鲡获得的胚胎发育过程的研究已报道,但人工诱导池塘养殖鳗鲡的胚胎及仔鱼早期发育未见报道。本文应用池塘养殖日本鳗鲡取代野生降海日本鳗鲡进行人工繁殖,2002年的孵化批次共80次,其中大部分为人工授精卵(60次),一部分(20次)为自然产卵。受精率为(44.03±21.99)%,孵化率为(65.76±19.48)%,共计获得苗种349.9万尾。连续记录了鳗鲡的胚胎和早期仔鱼的发育过程。在水温为20.5℃时,胚胎发育需要时间为49h,所需积温1005℃.h;在水温为22.5℃时,胚胎发育需要时间为39h,所需积温878℃.h;在水温为24.5℃时,胚胎发育需要时间为34h,所需积温833℃.h。     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.     

短短芽胞杆菌FJAT-0809-GLX几丁质酶基因(chiD)的克隆与原核表达

以短短芽胞杆菌FJAT-0809-GLX基因组DNA为模板,通过PCR扩增得到几丁质酶基因chiD序列,再将chiD序列连接到pMD18-T克隆载体上,形成重组载体pMD18-T-chiD,转化大肠杆菌DH5ɑ并测序,经过核苷酸和氨基酸序列分析获得几丁质酶基因chiD的序列片段为1 524 bp,编码507个氨基酸,理论蛋白质分子量约54.55 ku,其等电点约为5.77,表明该几丁质酶为酸性几丁质酶。将重组质粒pMD18-T-chiD双酶切后与表达载体pET-28a连接,构建重组表达载体pET28a-chiD,并转入大肠杆菌BL21中进行IPTG诱导表达,结果表明:重组蛋白质的分子量约55 ku,与预测的蛋白分子量结果一致。为了提高重组蛋白的表达量,对培养时间、IPTG浓度和温度3个参数进行优化,并将表达产物进行SDS-PAGE分析,最后得出重组载体pET28a-chiD的最佳诱导表达参数分别为8 h、0.5 mmol/L和28℃。这为短短芽胞杆菌来源的几丁质酶的进一步研究奠定了良好基础。     

弄岗马兜铃组培苗的生根培养

对弄岗马兜铃组培苗进行生根诱导研究,以达到可移栽的目的。在1/2改良MS固体培养基(KH2PO4 170 mg/L、蔗糖30 g/L,pH 5.8)中添加不同浓度的6-苄胺基腺嘌呤(6-BA)和吲哚丁酸(IBA)筛选最佳生根培养基配方。结果表明：在1/2改良MS固体培养基(蔗糖30 g/L,pH 5.8)加入1 mg/L的IBA具有较好的生根诱导作用,而6-BA对弄岗马兜铃组培苗不具有生根作用。     

普通小麦闭颖特性遗传分析及离体培养特性研究

为了给闭颖小麦的分子育种提供理论依据,进行了闭颖小麦闭颖授粉特性的遗传分析,同时对6个闭颖小麦系的花药、幼胚及成熟胚进行了离体培养特性的比较研究。结果表明,5组开、闭颖小麦杂交F1代个体全表现为开颖授粉,F2代个体开、闭颖分离,经卡平方适合性测验,开、闭颖株数均符合3∶1分离比例,表明小麦闭颖变异材料的闭颖授粉特性受一对隐性基因控制;不同闭颖小麦系离体愈伤组织诱导与再生特性有一定差异,供试的6个闭颖系中,133-1和133-8的离体培养特性优良,其中,133-1的成熟胚愈伤组织分化率最高,达到27.93%,133-8的幼胚分化率最高,达到40.70%,两材料的花药分化率也最好,都为4.48%,与对照材料小偃22处于同一水平;133-A的离体组织培养特性最差;3种外植体离体培养中,幼胚的培养再生效率最高,成熟胚次之,花药最差,供试材料的出愈率与分化率之间不具备明显的相关关系。     

白木香2种外植体的组织培养

沉香是国际上极负盛名的药用和香料资源之一。在亚洲许多国家,沉香产业既是传统行业也是发展迅猛的新兴产业。沉香优良种质的大规模应用对沉香产业的可持续发展具有重大意义。因此优良沉香种质资源是近年沉香产业关注的重点和热点,但一些优良种质的品质特性无法通过有性繁殖遗传,大规模繁育存在技术障碍。植物组织培养技术在品种优良性状保持、种子资源保存、快速繁殖等方面具有非常突出的优越性,被广泛应用于中药资源保护和中药产业发展。为建立工业化生产的沉香广谱再生体系,本文以白木香无菌苗和成龄植株的枝条为材料从外植体消毒、丛生芽诱导和生根培养三方面进行组织培养研究。结果表明,通过0.1%多菌灵溶液浸泡3 min,流动水冲洗3 h处理,对采摘自大田的白木香枝条有较好的除菌效果。0.1%升汞溶液消毒6~8 min,可有效保持外植体的成活率。在培养基中适当添加灭菌剂也能有效控制材料染菌。无菌苗茎段丛生芽诱导较易,大田枝条的丛生芽诱导较难。WPM + 20 g/L蔗糖+0.5 mg/L 6-BA+0.2 mg/L NAA (G₄)有利于无菌苗的丛生芽诱导;1/2 MS+25 g蔗糖+2 mg/L 6-BA+0.01 mg/L NAA (I₃),1/4 MS+20 g/L蔗糖+ 2 mg/L 6-BA+0.5 mg/L NAA (J₂)和WPM+20 g蔗糖+1 mg/L 6-BA+0.01 mg/L NAA (K₂)有利于成龄植株枝条外植体的丛生芽诱导,将膨大的丛生芽团切割后转接于不含植物生长调节剂的WPM培养基中可促进丛生芽伸长。WPM培养基中添加10~20 g/L蔗糖,并加入0.1~0.5 mg/L NAA可诱导无菌苗和茎段外植体的新芽生根。本研究以沉香2种外植体进行组织培养,成功获得再生植株,为沉香优良种质的无性繁殖提供技术参考。     

外源激素对桑树组培无菌苗继代增殖及生根的影响

[目的]探讨外源激素因子对桑树组培无菌苗继代增殖及生根的影响。[方法]以桂桑优12号为材料,从外源激素的不同种类、浓度及组合对桑树组培无菌苗继代增殖和生根诱导进行比较试验。[结果]以IBA 0.1 mg/L＋6-BA 1.5 mg/L＋TDZ 0.03 mg/L激素组合对芽增殖效果最好,芽增殖倍数达到5.28。 NAA对生根诱导的效果优于 IBA和 IAA;以 NAA 2.0 mg/L＋PP331.0 mg/L的激素组合对生根诱导效果最佳,生根率达100%,根数为7.01条,根长为1.38 cm。[结论]该研究为桑树种苗的离体培养工厂化生产提供技术参考。