氟喹诺酮类药物残留免疫学检测方法的研究

以沙拉沙星(SALX)为半抗原,牛血清白蛋白(BSA)偶联后免疫兔,获得对多种氟喹酮类(FQNS)药物有特异性的广谱性抗体.以SALX与卵清白蛋白(OVA)的偶联物作包被原,建立并优化了间接竞争ELISA检测方法,对SALX最低检出限达19.319 μg/L,且对诺氟沙星、环丙沙星和恩诺沙星均有特异性识别,最低检测限分别是22.646、26.181和19.054 μg/L.结果表明,该抗体可用于多种氟喹诺酮类药物残留筛选检测.     

Role of ghrelin in regulating rabbit ovarian function and the response to LH and IGF-I

The aim of these in vivo and in vitro studies was to examine the role of ghrelin in the control of plasma hormone concentrations, the proliferation, apoptosis and secretory activity of ovarian granulosa cells and the response of these cells to hormonal treatments. Female rabbits were injected with ghrelin (10 μg/animal/day for one week before ovulation induced by 25 IU PMSG and 0.25 IU LHRH). On the day of ovulation, blood samples were collected and analyzed for concentrations of progesterone (P₄), testosterone (T), estradiol (E₂), estrone-sulphate (ES), insulin-like growth factor I (IGF-I) and leptin (L) by RIA. Some control and ghrelin-treated animals were killed in the periovulatory period, their ovaries were weighed and granulosa cells were isolated and cultured for 2 d. Cell proliferation (expression of PCNA) and apoptosis (expression of TdT) were evaluated by immunocytochemistry and TUNEL respectively. Secretion of P₄, T, E₂, IGF-I, and prostaglandin F (PGF) by granulosa cells cultured with and without LH or IGF-I (1, 10 or 100 ng/ml medium) was assessed by RIA. The remaining control and treated animals were kept until parturition, while the number, viability and body weight of pups were recorded.     

猪圆环病毒2型Cap蛋白间接ELISA诊断方法的建立和应用

以纯化的原核表达的猪圆环病毒2型（PCV-2）重组结构蛋白（rCap蛋白）为包被抗原,建立PCV-2间接ELISA诊断方法;对抗原最适包被浓度、待检血清稀释浓度、重组抗原包被条件、封闭液的筛选、阴阳性临界值等条件进行了优化;对优化后的ELISA检测方法进行特异性试验、临床应用试验;组装试剂盒并进行试剂盒保存期试验。优化反应条件后确定的抗原最适包被浓度为2.815 μg/mL,抗原最佳包被条件为37 ℃ 2 h;血清最适稀释度为1∶40,酶标抗体最适稀释度为1∶500,最佳封闭液为1% BSA;阴阳性临界值判定标准为D450 nm≥0.33,判为阳性,D450 nm＜0.33,判为阴性。特异性试验结果表明,只有PCV-2阳性血清反应后的D450 nm＞0.33,包括抗PCV-1阳性血清在内的其他5种抗血清反应后的D450 nm＜0.33,说明所建立的ELISA方法具有良好的特异性。经对临床疑似PCV-2感染的100份血清样品进行检测,阳性检出率为69%。组装试剂盒在4 ℃条件下至少可保存12个月以上,说明所建立的诊断方法可以在生产中大量推广应用。本研究成功建立了能特异性检测抗PCV-2血清抗体的ELISA检测方法。     

The effects analysis of two neonicotinoid insecticides on in vitro maturation of porcine oocytes using hanging drop monoculture method

Acetamiprid (ACE) and imidacroprid (IMI) are known neonicotinoid insecticides with strong affinities for the insect-selective nicotinic acetylcholine receptor. These provide insect control by hyperstimulating insect nerves and are used for agricultural pest management. However, it has also been reported that ACE and IMI affect mammalian reproductive function. We determined the effects of ACE and IMI on the in vitro maturation of porcine oocytes. Significant decreases in nuclear maturation rates were observed in the ACE or IMI-exposed groups. Also, in matured oocytes from the ACE or IMI-exposed groups, irregular chromosomes were observed. Our results suggest that ACE and IMI exposure was detrimental to porcine oocytes and the extent of the effects depends on the concentration of exposure.     

Alteration of somatostatin receptor 2 expression in canine mammary gland tumor

Somatostatin receptor 2 (SSTR2) is a negative regulator of cell proliferation in human breast cancer. Since there is little information about SSTR2 in canine mammary gland tumor (MGT), we clarified its distribution and expression level in normal mammary gland, benign MGT and malignant MGT. SSTR2 expression determined by immunohistochemical staining was observed in the cytoplasm of luminal epithelial cells. The intensity was negatively correlated with malignancy: normal tissues and some of the benign tumors had the highest levels, while the malignant tumors had little or no SSTR2 expression. As for the Western blotting, SSTR2 protein level in benign tumors was significantly lower than the normal mammary gland. On the other hand, SSTR2 protein levels in two of three malignant tumors were higher than the other groups. These results suggest that SSTR2 expression alters according to the malignancy of canine MGT.     

几种重金属离子对高羊茅种子萌发及生理活性的响应

试验以高羊茅Festuca arundinacea"凌志1号"种子为材料,研究了镉(Cd2 )、铜(Cu2 )、汞(Hg2 )对种子萌发和幼芽细胞膜透性以及其体内过氧化氢酶(CAT)活性的影响.结果表明,0.5和5mg/L Hg2 极显著地促进萌发(P<0.01),50mg/L Cu2 极显著地抑制萌发(P<0.01),500mg/L各离子均显著抑制种子萌发(P<0.05).随处理的质量浓度逐渐增加,幼芽细胞膜透性逐渐增加.0.5mg/L的质量浓度下,Cu2 和Hg2 处理幼芽细胞膜透性显著低于对照(P<0.05);5和50mg/L的质量浓度下,Hg2 处理细胞膜透性最大;500mg/L的质量浓度下,Cu2 处理细胞膜透性最高.幼芽CAT的影响均表现为随质量浓度增加而逐渐升高,在0.5mg/L的质量浓度下,Cu2 处理CAT活性相对较高;在5mg/L的质量浓度下,Hg2 处理CAT活性相对较高;在50mg/L的质量浓度下,Cu2 和Hg2 处理CAT活性相对高于Cd2 处理;在500mg/L的质量的浓度下,Cu2 处理CAT活性相对较高.发芽率与细胞膜透性间存在较强的相关关系,发芽率和细胞膜透性与CAT呈负相关.综合分析几种重金属离子对高羊茅种子萌发的影响,Cd2 对高羊茅的毒害作用最大,Cu2 最小.     

印度梨形孢真菌对干旱胁迫下紫花苜蓿生长及抗旱性的影响

以紫花苜蓿为材料,通过盆栽砂培的方法,设置接种(40 mL 含有印度梨形孢菌丝体的菌剂)+干旱(土壤含水量为田间最大持水量的15%~20%)、未接种(40 mL无菌水)+干旱、接种+正常供水(土壤含水量保持在田间持水量的75%~80%)、未接种+正常供水4个处理,研究印度梨形孢真菌与紫花苜蓿共生对干旱胁迫下紫花苜蓿生长及抗旱性的影响。结果表明,印度梨形孢真菌在紫花苜蓿根部定殖率较高,达87.4%;与未接种+干旱处理相比,接种印度梨形孢对干旱胁迫下紫花苜蓿地上部和根系有明显影响,其中,地上部鲜重、干重、叶绿素含量、叶片相对含水量及叶片数分别比未接种处理显著增加了63.4%,69.2%,12.5%,17.1%和5.7片,根系鲜重、干重、主根长及侧根数分别增加了33.3%,57.1%,5.1 cm和5条(P<0.05);与未接种+干旱处理相比,接种后的紫花苜蓿叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、脯氨酸(Pro)和可溶性糖(SS)含量明显增强,分别是未接种处理的1.71,1.27,1.22,1.49和1.48倍(P<0.05),而超氧阴离子()与丙二醛(MDA)含量大幅度降低。因此,印度梨形孢真菌与紫花苜蓿共生可以促进干旱胁迫下紫花苜蓿生长,主要是通过刺激抗氧化酶活性和渗透调节物质的累积来抵抗干旱逆境。而在正常供水条件下,接种印度梨形孢没有明显的促进生长的作用。     

外源一氧化氮(NO)对低温胁迫下南瓜幼苗氧化损伤的保护效应

探讨外源 NO 供体硝普钠(SNP)对冷害胁迫下南瓜生长和氧化损伤的内在机制。以南瓜银辉2号和青栗为材料,通过室内人工模拟低温逆境的方法,研究 SNP(100 μmol/L)对冷害胁迫下南瓜幼苗生长、叶绿体色素含量、有机渗透调节物质含量及活性氧代谢的影响。结果表明,冷害导致南瓜幼苗活性氧积累增加,膜质过氧化加剧,光合色素含量下降,渗透调节能力降低,从而显著抑制南瓜幼苗的生长;正常生长条件下, SNP显著提高了超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性,显著增加幼苗地上和地下部干重及谷胱甘肽(GSH)、过氧化氢(H₂O₂)、类胡萝卜素和氨基酸总量;8 ℃/5 ℃(昼/夜)的低温胁迫下,叶面喷施外源100 μmol/L NO供体硝普钠(SNP),促进了植株生长和干物质积累,显著提高了叶片SOD、POD、APX 和GR的活性以及Pro和可溶性糖含量,减少了H₂O₂和膜脂过氧化产物丙二醛(MDA)的积累;外源NO明显提高了南瓜叶片叶绿素、类胡萝卜素含量、抗坏血酸(ASA)、GSH和可溶性蛋白及氨基酸总量。低温胁迫下,外源NO通过促进抗氧化酶活性、抗氧化剂及有机渗透调节物质含量的提高,降低H₂O₂和MDA的积累,保护了细胞膜结构的稳定性,维持了冷害下南瓜幼苗碳氮代谢的正常进行,增强了南瓜的抗冷性。试验条件下,NO对银辉2号的促进作用大于青栗。     

不同蛋白质源对草鱼和花鲈肉质影响的比较研究

本试验旨在比较不同蛋白质源对草鱼(Ctenopharyngodon idellus)及花鲈(Lateolabrax japonicas)体成分、肌肉游离氨基酸组成及鱼片质构特性的影响。设计草鱼和花鲈饲料各3种,分别等氮等能。3种草鱼试验饲料分别为以鱼粉为唯一蛋白质源的CI-FM组(对照组),以混合植物蛋白质(豆粕和大豆浓缩蛋白)替代80%鱼粉的CI-PPB80组及以混合植物蛋白质替代100%鱼粉的CI-PPB100组。3种花鲈试验饲料分别为以鱼粉为唯一蛋白质源的LJ-FM组(对照组),以混合植物蛋白质(棉籽浓缩蛋白和大豆浓缩蛋白)替代50%鱼粉的LJ-PPB50组和以混合植物蛋白质替代100%鱼粉的LJ-PPB100组。草鱼初始体重为(153.40±0.30)g,随机分为3组,每组3个重复,每个重复20尾鱼;花鲈初始体重为(12.97±0.03)g,随机分为3组,每组4个重复,每个重复25尾鱼。饲喂8周后,检测2种试验鱼的体成分、肌肉游离氨基酸含量及鱼片质构特性、剪切力、滴水损失及胶原蛋白含量。结果表明:与CI-FM组相比,CI-PPB100组草鱼肌肉粗脂肪含量显著降低(P0.05),粗灰分含量显著升高(P0.05);而花鲈肌肉粗脂肪含量则随替代水平的升高呈现先上升后下降的趋势,表现为LJ-PPB50组显著高于LJ-PPB100组(P0.05)。草鱼肌肉总游离氨基酸及生物胺前体含量各组间无显著差异(P0.05);随着替代水平的升高,花鲈肌肉总游离氨基酸及呈味氨基酸含量显著降低(P0.05),生物胺前体含量显著升高(P0.05)。2种试验鱼肌肉中存在各自的特异性成分。2种试验鱼生、熟鱼片的质构特性差异较大。草鱼的CI-FM组和花鲈的LJ-FM组生鱼片的硬度、黏性、咀嚼力、回弹力及剪切力均分别显著高于同鱼种的其他2组(P0.05)。对于熟鱼片,草鱼的CI-PPB80组内聚力显著高于其他2组(P0.05),其他各项指标各组间无显著差异(P0.05);花鲈的LJ-PPB100组硬度及黏附性显著高于LJ-FM组(P0.05),其他各项指标各组间无显著差异(P0.05)。根据结果,混合植物蛋白质替代鱼粉后降低了草鱼和花鲈鱼体脂肪的蓄积,对草鱼肌肉总游离氨基酸含量不造成影响,使花鲈肌肉呈味氨基酸含量下降,必需氨基酸含量减少,生物胺前体含量升高,货架期缩短;高植物蛋白质源饲料使得草鱼和花鲈的肉质均出现明显下降,而高鱼粉饲料可以保证2种试验鱼有较高的肉质。     

禽白血病病毒J亚群(ALV-J)的攻毒试验

将禽白血病病毒 J亚群 (AL V- J)内蒙株 NM876 1和 NM9996人工接种于 1日龄爱维茵肉种鸡 ,通过眼观、病理组织学检查观察了攻毒鸡的病理学特征 ;通过 PCR和 EL ISA技术检测了病毒感染率、抗体变化规律以及病毒和抗体的相关性。结果显示 ,攻毒鸡从第 3周开始即可检出病毒 ,NM876 1株和 NM9996株病毒感染率分别为 71.4 %和6 4 .3% ;从第 5周开始 ,出现较明显的病理学变化 ,病变特征以骨髓、肝脏、心脏、脾、卵巢等组织内髓细胞增生为主 ,而法氏囊、脑、坐骨神经无变化 ;攻毒鸡抗体消长变化有一定的规律 ,一般在 7～ 8周龄和 18～ 2 0周龄时分别出现 1次高峰 ,而在 4～ 6周龄、10周龄和 2 3周龄分别出现 1次低谷 ,提示鸡场进行 EL ISA检测时要避开这一时期 ;攻毒鸡产生耐受性病毒血症 ,即病毒阳性而抗体阴性 (V A- )的比例较高 (7/14 ,9/14 )。通过以上的研究证明 ,AL V- J内蒙株疾病模型复制成功 ,NM876 1、NM9996可作为原型株进行相关研究