青贮用乳酸菌的分离鉴定及生物学特性评价

本研究旨在从构树青贮、玉米青贮及饲喂构树青贮的羔羊粪便中分离鉴定可饲用乳酸菌,为开发构树青贮用乳酸菌提供试验材料。以构树青贮、玉米青贮以及饲喂构树青贮的羔羊粪便为菌株分离来源,使用含有1%碳酸钙的MRS琼脂培养基分离纯化乳酸菌,获得菌株经16S rRNA测序鉴定后对其中可饲用菌株进行生长、产酸性能的筛选,评估在pH 3.0、3.5、4.0条件下存活率和对大肠杆菌、金黄色葡萄球菌和沙门氏菌的抑制能力,并对筛选菌株进行菌株之间拮抗性测定。结果表明：获得可饲用乳酸菌共37株,包括植物乳杆菌(Lactobacillus plantarum)、布氏乳杆菌(Lactobacillus buchneri)、副干酪乳杆菌(Lactobacillus paracasei)、粪肠球菌(Enterococcus faecalis)4种,不同的菌株在生长和产酸性能上表现出差异,在不同pH中存活率和抑菌性能也具有菌株特异性。综合比较不同菌株之间的生物学特性,植物乳杆菌GQ-3-2、GQ-3-29及GQ-5-4具有良好的生长、产酸及抑菌性能;粪肠球菌Y-3-15在发酵2 h进入对数生长期,能够为植物乳杆菌的大量繁...     

内蒙古通辽市牧区传统乳制品中乳杆菌的分离鉴定

试验从内蒙古通辽市科左后旗和扎鲁特旗牧区自产的乳样中分离到29株乳杆前,并对其进行了鉴定。结果显示．优势菌株植物乳杆菌21株,玉米乳杆菌、干酪乳杆菌、肠孔杆菌、短乳杆菌和嗜酸乳杆菌各1株,另有3株没有鉴定到种。     

呼伦贝尔草原野生牧草青贮中优良乳酸菌的分离及鉴定

本研究对呼伦贝尔草原牧草青贮饲料中的乳酸菌进行了分离、鉴定,旨为筛选出优良乳酸菌菌株。以4个地区不同群落牧草的青贮饲料为试验材料,并采用传统微生物学鉴定方法及16S rRNA序列分析对分离得到的乳酸菌菌株进行了鉴定。菌株DME53为草乳杆菌,DMG108为干酪乳杆菌,DMC80为植物乳杆菌,DMG138为短乳杆菌,DMG139为戊糖片球菌;其中菌株DMG138为异型发酵乳酸菌,其余4株菌均为同型发酵乳酸菌。除DMG139在40和45 ℃下生长微弱外,其余菌株均在5~45 ℃不同温度条件下, 3.0%和6.5% NaCl培养液中良好生长。除菌株DME53在pH 3.0下不能生长外,其余菌株均在pH 3.0~8.0条件下良好生长或微弱生长。菌株DMC80具有产酸能力较强、发酵速率较快、耐酸、耐低温等特性,可作为制备适用于呼伦贝尔地区青贮饲料菌制剂的优良菌株。     

黏附与聚集、抗肿瘤及抗菌特性乳杆菌的筛选

为获得具有高黏附与聚集、抗肿瘤及抗菌特性的益生乳杆菌,本研究以10株经16s r DNA鉴定的乳杆菌为候选菌,以结肠癌HT-29细胞为体外模型研究其黏附能力和抗肿瘤特性及其黏附能力与自聚集能力的相关性。将乳杆菌与致病性大肠埃希氏菌、鸡白痢沙门氏菌及金黄色葡萄球菌分别共孵育,探究了乳杆菌的共聚集能力与抗菌能力。结果显示:有6株乳杆菌的自聚集能力在20%以上;4株乳杆菌粘附内化HT-29细胞的内化率超过5%,经荧光显微镜观察,菌株Lac.99对HT-29细胞具有最强的黏附特性;菌株Lac.117和Lac.56对HT-29细胞的生长具有较好的抑制作用;4株乳杆菌对指示致病菌具有较强的拮抗作用,其中Lac.117菌株作用最强。综上所述,候选菌株中Lac.56在黏附、抗菌和抗肿瘤方面均表现出良好的益生特性,本实验为进一步研究和开发具有潜在应用价值的益生菌制剂提供了依据。     

干酪生产用菌种的筛选

采用从新疆民族乳制品中分离的菌株，研究了三种单一菌株（L1乳酸乳球菌、L2瑞士乳杆菌、L3干酪乳杆菌）和四种混合菌株（L4乳酸乳球菌：瑞士乳杆菌（1：1）、L5乳酸乳球菌：干酪乳杆菌（1：1）、L6瑞士乳杆菌：干酪乳杆菌（1：1）、L7嗜热链球菌：保加利亚乳杆菌（1：1））的发酵性能和生化特性，确定了适于干酪加工的菌种为L1、L5、L6。     

一株布氏乳杆菌的分离鉴定

研究从发酵秸秆中分离获得1株抑菌性能较强的乳酸菌(BS6),并通过形态学、生理生化和分子鉴定对该菌株进行鉴定,最终确定菌株BS6为布氏乳杆菌,并对布氏乳杆菌BS6的生长曲线、产酸曲线及耐酸性能进行研究,结果表明,菌株BS6生长速率快、产酸性能强且能够耐受pH环境,为青贮饲料接种剂提供有力的理论依据。     

一株新饲用乳杆菌的分离鉴定及益生性研究

从猪粪便中分离得到一株新饲用乳杆菌,对其16S rDNA基因进行了测序,该段基因序列已提交GenBank,登录号为HQ641209。将该菌株16S rDNA基因序列进行在线BLAST比对并与其他乳杆菌属菌株的16S rDNA序列建立系统发育树,结果显示该菌16S rDNA序列与NCBI公布的约氏乳杆菌Lactobacillus johnsonii (AB295648)的同源性为99.7%,因此该菌鉴定为约氏乳杆菌(Lactobacillus johnsonii),将其命名为"Lactobacillus johnsonii Sun001"。经测试,该菌株对E.coli K88、K99菌株的生长有抑制作用,在pH为2.5的模拟胃液中处理3 h存活率为68.8%,在0.3%胆盐浓度的模拟肠液中处理3 h存活率为21.4%。     

鸡源耐镉乳酸菌株的分离与鉴定

本试验旨在从鸡肠道中分离筛选出耐镉(Cd)的乳酸菌,为将其开发应用于防控家禽饲料重金属Cd污染提供理论基础。取鸡肠道内容物的样品进行乳酸菌分离,以耐受100 mg/L的Cd为标准筛选乳酸菌,采用分子生物学方法鉴定菌株。对筛选出的乳酸菌株进行生长性能、耐Cd性能及Cd吸附性能等测定。结果表明：本试验共筛选出5株耐Cd乳酸菌,分别为1株约氏乳杆菌Cd3-1和4株卷曲乳杆菌Cd1-1、Cd5-2、Cd4-5和Cd8-2。其中Cd3-1、Cd5-2和Cd8-2菌株最高可耐受1 000 mg/L的Cd,而Cd1-1和Cd4-5菌株最高可耐受400 mg/L的Cd。各菌株对50 mg/L Cd的吸附率随溶液pH的升高而增加,在pH为6.0和7.0时Cd的吸附率均显著高于pH为2.6时Cd的吸附率(P<0.05)。5株乳酸菌均可耐受0.3%的胆盐,且可抑制金黄色葡萄球菌的生长。综合上述结果,本试验从鸡肠道中分离筛选鉴定出5株耐Cd乳酸菌,包括1株约氏乳杆菌和4株卷曲乳杆菌,在pH接近中性的条件下,对Cd的吸附率可达30%。     

高寒地区菊芋青贮物料耐低温乳酸菌的筛选与鉴定

为筛选可提升高寒地区菊芋(Helianthus tuberosus)青贮发酵品质的耐低温乳酸菌,本研究以‘青芋2号’菊芋青贮物料为材料,利用平板培养法分离乳酸菌株,分析菌株生理生化特性,并通过16S rRNA测序进行菌株种属鉴定。结果表明,菊芋青贮物料中共分离得到34株乳酸菌株,经低温及生理生化筛选获得13株耐低温乳酸菌株,包括11株同型发酵乳酸菌和2株异型发酵乳酸菌,其耐酸性和耐盐性均较强;经产酸和生长能力对比,GN02菌株生长最快(OD=2.63),GN10菌株具有更强的产酸能力(pH=3.59),XN25菌株兼具较强的生长和产酸能力;16S rRNA测序鉴定出9株植物乳杆菌(Lactiplantibacillus plantarum)、1株戊糖乳杆菌(Lactiplantibacillus pentosus)、1株棒状乳杆菌(Lactiplantibacillus coryniformis)、1株短乳杆菌(Lactiplantibacillus brevis)及1株无法鉴定。本研究筛选出适合高寒地区菊芋青贮发酵的耐低温植物乳杆菌、戊糖乳杆菌、短乳杆菌各1株。     

东北虎肠道降胆固醇乳杆菌的分离鉴定

为了筛选适宜开发降胆固醇保健食品的乳杆菌,试验采用乳杆菌选择培养基(LBS),从东北虎肠道分离、筛选得到10株乳杆菌。采用高胆固醇乳酸杆菌培养液(MRSO-CHOL)对分离菌进行体外降胆固醇试验,并对降胆固醇最高的3株菌进行耐酸及耐胆汁盐试验;对胆固醇降解率高、耐酸、耐胆汁盐的菌株进行形态学、生理生化反应和16S rDNA鉴定。结果表明:从东北虎肠道分离得到的10株乳杆菌,体外均有降解胆固醇的能力,降解率为25.00%～65.00%,平均降解率为45.00%。菌株1胆固醇降解率为55.00%,且耐酸、耐胆汁盐的能力均较强,鉴定其为嗜酸乳杆菌。     

原料乳中耐冷假单胞菌的种类及其肽酶活力

在冷藏的原料乳中,假单胞菌经常出现并成为主导菌群,其分泌的耐热酶会使原料乳变质。对从原料乳中分离的102株假单胞菌经rpoB基因测序进行菌种分析,用偶氮酪蛋白测试所有假单胞菌在2种培养条件(30℃培养2d或6℃培养8d)下胞外肽酶的蛋白水解活性和耐热性。结果表明:荧光假单胞菌(Pseudomonas fluorescens)是假单胞菌中最主要的菌种,其次是莓实假单胞菌(Pseudomonas fragi)和假单胞菌sp1 (Pseudomonas sp1)。不同假单胞菌菌种之间的产肽酶能力存在明显差异。大多数荧光假单胞菌(23/41)在不同温度(30℃和6℃)条件下培养后能产肽酶,而大多数的假单胞菌sp1不产肽酶,所有分离的莓实假单胞菌(13/13)只在6℃条件下产肽酶,大多数的假单胞菌所产肽酶都有热稳定性。在6℃培养条件下,荧光假单胞菌菌株M02有最高肽酶活力(11.78ΔA/ (h·mL)),其胞外肽酶最佳催化pH值为6.5,最佳催化温度为40℃。当M02的接种浓度为103 CFU/mL时,6℃培养5d就能使脱脂乳溶液产生沉淀。对耐冷假单胞菌的种类和产肽酶能力进行分析和考察有助于提高原料乳或乳制品的质量和安全性。     

多年生黑麦草愈伤组织航天搭载后再生株系的抗旱性研究

利用“神舟七号”飞船搭载多年生黑麦草品种(Derby)的胚性愈伤组织经空间诱变后,从搭载再生株系(SP)群体中获得了3个抗旱变异株系。自然干旱条件下,对筛选出的3个抗旱变异株系进行了生理生化分析,结果表明,随自然干旱时间的延长,3个抗旱变异株系均表现出叶片相对含水量和叶绿素含量下降、SOD活性先升后降、丙二醛含量及游离脯氨酸含量上升等特征;不同株系间各指标的变化幅度存在差异,通过上述5项指标综合评价后初步认为,3个抗旱变异株系的抗旱性顺序为SP-64>SP-119>SP-17,与表征永久萎蔫系数反映的结果一致,表明筛选出的抗旱变异株系具有抗旱生理基础。以上结果说明,空间诱变能诱发多年生黑麦草抗旱性的变异;获得的抗旱变异株系为选育抗旱草坪草品种提供了新的种质资源。     

Immunohistochemical localization of chromogranin A in normal tissues from laboratory animals

To analyze the distribution of Chromogranin A in endocrine cells of various species of laboratory animals (dog, gerbil, guinea pig, hamster, monkey, mouse, and fetal, neonatal, and adult rats), normal tissues were stained immunohistochemically with polyclonal anti-bovine Chromogranin A antiserum (SP-1). Selected tissues (pituitary, adrenal, thyroid, parathyroid, pancreas, brain, peripheral nerve, stomach, small and large intestine, bone marrow, spleen, thymus, lymph node, and liver) from these species and from the rabbit were stained with two monoclonal anti-human Chromogranin A antibodies (LK2H10 and PHE5) to compare the immunoreactivities of the monoclonal antibodies and polyclonal antiserum. Staining with the polyclonal antiserum (SP-1) resulted in a broader spectrum of immunoreactivity but had more nonspecific background staining than either monoclonal antibody. Immunoreactivity and staining intensity with SP-1 varied between species, but most endocrine tissues (pituitary cells in the anterior and intermediate lobes, thyroid "C" cells, adrenal medulla, parathyroid, pancreatic islets, and enterochromaffin cells) from most species stained positively. In some species, pancreatic alpha cells stained more intensely, and two populations of adrenal medullary cells with different staining intensities were observed. Sciatic nerve (axonal area) was immunoreactive with monoclonal antibodies and/or the polyclonal antiserum in several species. The spectrum of immunoreactive tissues from fetal and neonatal rats increased with age. There was good cross-reactivity between species with SP-1, but not with either LK2H10 or PHE5. These results indicate that many endocrine cells with secretory granules in laboratory animals express Chromogranin A and that a polyclonal antiserum, such as SP-1, is more sensitive in detecting this protein in various species than monoclonal antibodies such as LK2H10 or PHE5.     

铜绿假单胞菌弹性蛋白酶基因的克隆及在昆虫细胞中的表达

以产弹性蛋白酶的铜绿假单胞菌基因组DNA为模板,经PCR扩增得到约1.5 kb的弹性蛋白酶基因,克隆到质粒pET28 a,连同其上游的6×H is序列转移到pBacPAK8构建成pBacPAK8-H is-E la转移载体。采用L ipofec-tin法,将载体pBacPAK8-H is-E la和亲本病毒BacPAK6共转染Sf21细胞,经空斑纯化分离得到重组病毒BacPAK6-E la。通过双酶切及单酶切鉴定证实弹性蛋白酶基因已插入到病毒基因组的多角体启动子下游,SDS-PAGE检测到在昆虫细胞中表达约34 kD的蛋白条带,以弹性蛋白为底物的生化反应证明了表达产物具有一定的生物学活性。研究结果为弹性蛋白酶的制备提供了新的方法。     

Localization and Coexistence of Calcium-Binding Proteins and Neuropeptides in the Vagal Ganglia of the Goat

This study was performed to investigate the neurochemical characteristics of the vagal ganglia of the goat by immunohistochemical methods using calbindin D-28k (CB), calretinin (CR). parvalbumin (PA), substance P (SP). calcitonin generelated peptide (CGRP) and galanin (GAL) antibodies. In the proximal vagal ganglia (jugular ganglia), CGRP- (57.1%), SP- (48.2%), GAL- (8.6%), PA- (8.7%), CB- (8.5%) and CR-like (5.3%) immunoreactive cells were observed. In the distal vagal ganglia (nodose ganglia), CGRP- (40.5%), SP- (30.20%), CB- (22.0%) and CR-like (18.10%) immunoreactive cells were present. The double immunohistochemical study showed, that in the proximal vagal ganglia, CGRP immunoreactivity was co-localized in SP- (84.8%), GAL-(100%), CB- (5.6%) and CR- (5.7%) immunoreactive cells: SP immunoreactivity was co-localized in the CGRP- (80.0%), GAL- (100%). CB- (5.3%) and CR- (5.6%) immunoreactive cells; GAL immunoreactivity coexisted in the CGRP- (4.4%) and SP- (19.8%) immunoreactive cells, but not in calcium-binding proteins (CBP)-immunoreactive cells; PA immunoreactivity was absent in the CGRP- and SP-immunoreactive cells; CB and CR immunoreactivities were seen in the CGRP-(0.8%) and SP-immunoreactive (0.9%) cells. On the other hand, in the distal vagal ganglia, CGRP immunoreactivity appeared in SP- (66.6%), CB- (1.0%) and CR- (1.2%) immunoreactive cells; SP immunoreactivities were observed in the CGRP- (44.1%), CB- (1.0%) and CR- (1.2%) immunoreactive cells; CB immunoreactivities were present in the CGRP- (0.5%) and SP- (0.8%) immunoreactive cells; CR immunoreactivities were contained in the CGRP- (0.5%) and SP- (0.8%) immunoreactive cells. These findings indicate that the goat is distinct from other mammalian species in the distribution and localization of neurochemical substances in the vagal ganglia. and suggest that these differences may be related to physiological characteristics, particular those of the ruminant digestive system.     

Staphylococcal protein A (SpA): a potent in vivo chemotactic agent for rat leucocytes

Staphylococcal protein A had strong chemotactic attraction in vivo to rat leucocytes. Doses as small as 5 micrograms attracted net leucocytes into experimental pellets in 6 h. 50 micrograms Staphylococcal protein A showed maximum chemotactic activity and does greater or less than 50 micrograms attracted less net leucocytes into experimental pellets. The effect of time on the chemoattraction of 50 micrograms Staphylococcal protein A showed that it was an early chemoattractant. Chemotactic activity for this dose, shown by the chemotactic index, reached a peak at 6 h followed by maximum leucocytic infiltration, and almost disappeared completely at 12 h. Leucocytic migration into control pellets rose from 3 h and reached a peak at 12 h (later than the chemotactic peak). Staphylococcal protein A also showed in this study a "later reaction" from 24 to 36 h, resulting in local inflammation of the test site and rise in cellular response.     

Stress Protein Expression in Coho Salmon with Bacterial Kidney Disease

Abstract

The expression of stress protein-70 (SP-70), also known as heat shock protein-70 (HSP-70), was measured by enzyme-linked immunosorbent assay in kidney and liver from coho salmon Oncorhynchus kisutch with bacterial kidney disease (BKD) experimentally induced by injection with Renibacterium salmoninarum. Fish with BKD had more SP-70 in both kidney and liver than did the control fish. The SP-70 measured was derived from the host tissue, not from the pathogen. In fish without clinical disease, SP-70 was not significantly elevated. Elevated stress protein (SP) levels in fish with BKD raise the possibility that BKD or other infectious diseases may interfere with the use of SP induction as a marker of environmental stress in fish and lead to statistical artifacts when the prevalence of disease is not considered.     

肽在瘤胃内的代谢和利用

微生物蛋白降解作用形成的多肽经过一系列肽酶最终转化为氨基酸,其速度取决于多肽的化学组成和N端结构.大多数情况下,瘤胃肽在栖瘤胃拟杆菌分泌的二肽基肽酶作用下,从多肽N未端释放二肽而不是游离氨基酸.通过抑制栖瘤胃拟杆菌的二肽基肽酶活性或该菌对肽吸收率可以减慢混合瘤胃群落对肽的水解速率.肽和氨基酸刺激瘤胃微生物的生长,并且对许多种菌的最佳生长是必需的.纤维分解菌是个例外.当然,细菌蛋白质合成时利用氨与肽和氨基酸的比例也取决于它们的浓度.     

江河源区不同建植期人工草地土壤养分及微生物量磷和磷酸酶活性研究

研究了"黑土滩"退化草地上建植的2,4和6龄垂穗披碱草人工草地土壤养分、微生物量磷、中性磷酸酶活性变化以及它们之间的相互关系。结果表明,随建植期的增加,土壤pH值呈现先降低后升高的趋势,人工4龄草地土壤pH值最低,人工6龄草地最高。从时间尺度看,人工2龄草地土壤养分含量、土壤微生物量磷含量和土壤中性磷酸酶活性较"黑土滩"退化草地明显升高,随着建植期的增加,人工4龄草地各测定指标明显下降,而人工6龄草地各测定指标再次升高。4种草地土壤养分含量、微生物量磷含量和中性磷酸酶活性在土壤剖面中均呈现随土壤深度的增加而递减的趋势。相关分析结果表明,土壤微生物量磷与有机碳、全氮、速效氮和速效钾含量之间呈极显著正相关关系,与全磷含量间呈显著正相关关系;土壤中性磷酸酶活性与大多数土壤养分呈显著或极显著正相关关系,其中与土壤有机碳的相关系数最大;土壤微生物量磷与中性磷酸酶活性呈极显著正相关关系。     

气相色谱法测定动物肉中脂肪酸组成

该试验通过GC-2014气相色谱法,采用SP-2560毛细管气相色谱柱,利用外标法定量分析绒山羊肌肉中多种脂肪酸含量。结果表明,该方法简单、快速,第10碳以上长链脂肪酸的线性范围好,相关系数为0.9991以上,回收率在88.29%～96.21%之间,相对标准偏差在5%以内。