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1.
《黑龙江畜牧兽医》2020,(13)
为了从青海省寒旱环境的盐渍土、水、植物中筛选分离及鉴定可以生产脲酶的耐盐菌,改善青海牧区土壤环境,探索寒旱盐渍土环境中微生物矿化碳酸钙技术的可行性,试验利用5%盐浓度的筛选培养基筛选耐盐菌,从中筛选最优脲酶产生菌,测定其生长曲线和产酶曲线,并进行菌株鉴定及微生物矿化碳酸钙技术可行性测定。结果表明:筛选出200株耐盐菌,其中分离得到1株高产耐盐脲酶产生菌,该菌株在5%盐浓度下的液体培养基中于37℃、180 r/min振荡培养时,其对数生长期出现在4～24 h,最佳发酵时间为48 h,初始酶活性最高达到2.21 U/mL,经鉴定该菌株为木糖葡萄球菌,登录号为MN420830,生成的沉淀为碳酸钙。说明从青海省寒旱盐渍土环境中可以筛选出耐盐脲酶产生菌,且该菌在实验室环境条件下进行微生物矿化碳酸钙技术是可行的。 相似文献
2.
黄芪多糖对体外培养大鼠肠黏膜微血管内皮细胞增殖的影响 总被引：3，自引：2，他引：1
研究黄芪多糖对体外培养的大鼠肠黏膜微血管内皮细胞（rat intestinal mucosa microvascular endothelial cells,RIMMVECs）增殖的影响。应用不同浓度的黄芪多糖作用于体外培养的RIMMVECs。通过MTT法检测黄芪多糖对RIMMVECs增殖的影响。不同浓度的黄芪多糖对细胞增殖的影响不同:黄芪多糖浓度在25～800 mg/L时可明显促进细胞的增殖（P<0.05或0.01）;浓度为1.6×103 mg/L时对细胞的增殖无影响;浓度在3.2×103～25.6×103 mg/L时对细胞生长具有抑制作用（P<0.05或0.01）。黄芪多糖有促进体外培养RIMMVECs的增殖作用,此作用对保护内皮损伤、加速内皮修复有重要作用。 相似文献
3.
《饲料工业》2021,(5)
试验旨在从西农萨能奶山羊瘤胃液中分离出产蛋白酶的酵母菌株,并对其产酶能力进行测定。以蛋白酶的活性进行筛选,得到高产蛋白酶酵母菌,对其产酶条件进行优化。经形态学观察及26S rDNA序列分析鉴定出5株产蛋白酶酵母菌,分别对各菌株产酶条件进行优化,在最佳产酶条件下,菌株库德毕赤酵母Y1(时间48 h、接菌量3%、温度30℃、pH值9)产酶活力达229.89 U/mL,东方伊萨酵母Y5(时间48 h、接菌量4%、温度37℃、pH值9)产酶活力达212.63 U/mL,马克斯克鲁维酵母Y16(时间48 h、接菌量3%、温度37℃、pH值9)产酶活力达208.44 U/mL,近平滑假丝酵母Y20(时间48 h、接菌量4%、温度37℃、pH值9)产酶活力达211.74 U/mL,单孢酿酒酵母Y21(时间48 h、接菌量4%、温度30℃、pH值9)产酶活力达209.96 U/mL。同时对五株酵母菌产水解酶的效果进行了探究,发现库德毕赤酵母Y1、马克斯克鲁维酵母Y16、单孢酿酒酵母Y21具有较高的植酸酶活性,达到14.38、15.34、15.63 U/mL;五株菌株均表现出较高的果胶酶活性,其中单孢酿酒酵母Y21显著高于其他菌株(P0.05),达到15.18 U/mL;五株菌株均表现出较高的纤维素酶活性,均超过180 U/mL,其中近平滑假丝酵母Y20显著高于其他菌株(P0.05),达到199.08 U/mL。试验成功分离出5株产蛋白酶菌。库德毕赤酵母Y1、马克斯克鲁维酵母Y16、单孢酿酒酵母Y21具有较高的植酸酶活性。五株菌株均表现出较高的果胶酶活性、纤维素酶活性。 相似文献
4.
《畜牧与兽医》2015,(10):35-40
为筛选产漆酶的木质素降解菌,采用愈创木酚平板法和RB亮蓝平板法进行菌株筛选,利用16S r DNA序列分析对菌株进行鉴定,通过液体发酵培养对漆酶活性和木质素降解能力进行研究。将漆酶基因cot A克隆到表达载体p ET-32a(+)上获得重组质粒p ET-32a(+)-Cot A并转化到BL21中进行表达,通过酶活测定将表达条件进行优化。结果:从白蚁肠道中成功筛选出1株产漆酶的木质素降解菌,鉴定为枯草芽胞杆菌。漆酶的最适酶活温度为60℃,最适酶活p H为3.5~4.0;0.2 mmol/L Cu2+可诱导菌株漆酶的产生,缩短了产酶时间并提高了酶活性。菌株木质素降解率在液体发酵培养24 h后达到了17.1%。漆酶基因大小为1 542 bp,表达的重组蛋白为85 ku,属于有活性的可溶性蛋白。在IPTG浓度为1mmol/L、Cu2+浓度为0.3mmol/L、诱导时间为9 h的条件下,漆酶蛋白活性可达到113.10 U/L。研究表明,本试验成功筛选出1株降解木质素能力较强的枯草芽胞杆菌,获得可分泌较高活性漆酶的重组菌,为木质素酶在畜牧业生产实践上的应用奠定了基础。 相似文献
5.
为了提高纳豆芽孢杆菌的抑菌活性,筛选出高抑菌活性菌株,试验以纳豆芽孢杆菌BN-3作为始发菌,通过紫外线(UV)和硫酸二乙酯(DES)对其进行复合诱变,得到变异菌株XC-80,并进行营养源优化及发酵条件优化正交试验,使变异菌达到最高的抑菌活性。结果表明:紫外线最佳诱变时间为12 s,硫酸二乙酯最佳涂布浓度为0.3%,复合诱变使纳豆芽孢杆菌抑菌活性提高16.9%;经营养源优化,变异菌株XC-80最适碳源为葡萄糖,最适氮源为胰蛋白胨,最适金属盐为硫酸镁;正交试验得出优化发酵条件为葡萄糖2%,胰蛋白胨0.5%,硫酸镁0.06%,初始pH值7.2,接种量0.5%,培养温度30℃,转数140 r/min,培养时间72 h,抑菌活性较诱变后菌株再次提高25.2%;最终筛选后菌株较始发菌抑菌活性累计提高46.4%。说明复合诱变及发酵条件优化筛选出了具有高抑菌活性的变异菌株XC-80,优化了发酵条件,显著提高了发酵液抑菌活性。 相似文献
6.
选用从雷州半岛近岸海域分离鉴定的2株海洋红酵母（胶红酵母J6和黏红酵母J2）作为出发菌株,进行亚硝基胍（NTG）和紫外线（UV）复合诱变,以期筛选出高产类胡萝卜素的突变株。先用NTG处理出发菌株的菌悬液60 min,再用UV照射120 s,且要连续诱变3次;每次复合诱变后的菌悬液经短时间的液体培养后涂布于红酵母琼脂平板培养,从中筛选出一定数量的第1、2和3代突变菌株;将突变菌株接种于红酵母发酵培养基,经培养、离心和烘干,制备出干菌体细胞;用分光光度计法测定干菌体细胞中的类胡萝卜素含量,筛选出高产类胡萝卜素的突变株,并进行遗传稳定性试验。结果表明,胶红酵母J6的正向突变率为56.57%（56/99）,6株高产类胡萝卜素的突变株中,第3代3株,第2代2株,第1代1株,类胡萝卜素含量最高的突变株是胶红酵母J6-82,比出发菌株提高了99.56%;黏红酵母J2的正向突变率为53.01%（44/83）,6株高产类胡萝卜素的突变株中,第3代4株,第2代2株,类胡萝卜素含量最高的突变株是黏红酵母J2-75,比出发菌株提高了93.64%。本试验采用的NTG和UV 3次复合诱变的方法有利于海洋红酵母的变异及其类胡萝卜素含量的提升,且突变菌株有良好的遗传稳定性。 相似文献
7.
选用从雷州半岛近岸海域分离鉴定的2株海洋红酵母(胶红酵母J6和黏红酵母J2)作为出发菌株,进行亚硝基胍(NTG)和紫外线(UV)复合诱变,以期筛选出高产类胡萝卜素的突变株。先用NTG处理出发菌株的菌悬液60min,再用UV照射120s,且要连续诱变3次;每次复合诱变后的菌悬液经短时间的液体培养后涂布于红酵母琼脂平板培养,从中筛选出一定数量的第1、2和3代突变菌株;将突变菌株接种于红酵母发酵培养基,经培养、离心和烘干,制备出干菌体细胞;用分光光度计法测定干菌体细胞中的类胡萝卜素含量,筛选出高产类胡萝卜素的突变株,并进行遗传稳定性试验。结果表明,胶红酵母J6的正向突变率为56.57%(56/99),6株高产类胡萝卜素的突变株中,第3代3株,第2代2株,第1代1株,类胡萝卜素含量最高的突变株是胶红酵母J6-82,比出发菌株提高了99.56%;黏红酵母J2的正向突变率为53.01%(44/83),6株高产类胡萝卜素的突变株中,第3代4株,第2代2株,类胡萝卜素含量最高的突变株是黏红酵母J2-75,比出发菌株提高了93.64%。本试验采用的NTG和UV 3次复合诱变的方法有利于海洋红酵母的变异及其类胡萝卜素含量的提升,且突变菌株有良好的遗传稳定性。 相似文献
8.
减少瘤胃甲烷排放措施的研究已成为当前的研究热点之一。本研究以酒、面包和畜禽专用酿酒酵母产品为原材料,从中分离到三株酵母菌,将其制成冻干粉,通过体外产气试验,筛选出了一株抑制瘤胃甲烷排放效果最为显著的酵母菌株。然后以该酵母菌株作为研究对象,进行分子生物学鉴定;采用分光光度计比浊法测定该酵母菌株的生长曲线,确定菌体生长规律;并以此为依据,设计单因素试验考察该酵母菌株最适宜的生长条件。结果表明,分离得到的酵母菌株均为酿酒酵母saccharomyces cerevisiae;抑制瘤胃体外甲烷排放效果最显著的1株酵母菌分离自面包中;该酿酒酵母菌株的对数生长期为8～24h,其最佳培养条件为:生长温度30℃,pH 5.5,培养基中葡萄糖的浓度4.0%。 相似文献
9.
10.
本实验利用乳酸杆菌和酵母菌发酵泌乳母猪全价料,根据发酵前后饲料的pH值、活菌数、粗蛋白、总酸含量和酸溶蛋白含量等指标,筛选出适宜母乳母猪全价料发酵的乳酸杆菌和酵母菌;利用正交试验筛选了最优混菌发酵组合,并从混菌比例、水分、温度和发酵时间等关键因素研究对发酵饲料品质的影响,筛选出适宜乳酸杆菌和酵母菌混菌发酵的最优工艺。结果发现:适宜泌乳母猪全价料发酵的乳酸杆菌为BLCC2-0015,在发酵48 h时,pH值达到4.12,活菌数为60×10~8CFU/g,总酸含量高达22.56 mg/g;酵母菌为BLCC4-0021,在发酵48 h时活菌数为44.50×10~8CFU/g,粗蛋白含量高出对照组40.69%;两者复配发酵最佳工艺为接种比例为BLCC2-0015:BLCC4-0021=1.5%∶1.0%、料水比为1∶0.6、发酵温度为32℃、发酵时间48 h,在此条件下,乳酸杆菌活菌数为74.53×10~8CFU/g,酵母菌活菌数为12.04×10~8cfu/g,总酸含量为18.65 mg/g,酸溶蛋白含量为17.32%。 相似文献
11.
García-Hernández Y Rodríguez Z Brandão LR Rosa CA Nicoli JR Elías Iglesias A Peréz-Sanchez T Salabarría RB Halaihel N 《Research in veterinary science》2012,93(2):798-802
The objective of this study was to isolate and identify yeast strains from broilers excreta and to evaluate in vitro their potential for probiotic use in animal production.Methods and resultsNine yeast strains were isolated and presumptively pre-identified by biochemical assays. These isolates were grouped in six different molecular profiles using PCR-fingerprinting technique. Each profile was identified by sequencing of the D1/D2 domains of the large subunit of the 26S rRNA gene. These yeasts were identified as: Trichosporon sp. (LV-2), Wickerhamomyces anomalus (LV-6), Pichia kudriavzevii (LV-8), Kodamaea ohmeri (LV-9) and Trichosporon asahii (LV-10). A pre-screening of the strains for probiotic use was based on their ability to agglutinate pathogenic micro-organisms. These yeast strains were characterized for specific growth rate, duplication time, their cell surface hydrophobicity, medium acidification, resistance to low pH (2.0, 2.5 and 3.0) and concentrations of bile salts (0.3% and 0.6%). The isolate of W. anomalus (LV-6) had the highest agglutinating and adherence capacity, a growth rate of 2.07 × 108 cfu/mL in 24 h at 30 °C, decreasing the medium pH from 6.5 to 5.23, a 25% hydrophobicity and an elevated capacity to grow under stress conditions.ConclusionsW. anomalus strain LV-6 showed the best characteristics for use as a probiotic candidate.Significance and impact of the Study:The data from this study helped in choosing a probiotic candidate from yeast to use in broiler production. 相似文献
12.
Ryoka MIYAKE Takakaki ITO Hiroshi KAMATA Rui KANO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(7):1090
We isolated ascomycetous yeasts including Candida species, that originally belonged to the genus Candida, from bulk milk in the Aichi area of Japan, and determined the minimum inhibitory concentrations (MICs) of antifungal drugs on these isolates by conducting E-tests. We isolated 7 human pathogenic species (14 isolates) from 14 bulk milk samples: 5 Candida species of yeasts, and 2 Candida-related species. Two isolates of C. albicans and C. inconspicua were resistant to fluconazole (MIC >32 mg/l). One isolate of C. krusei was resistant to both azoles (fluconazole: >256 mg/ml and itraconazole: 4 mg/l). One isolate of C. catenulata might be resistant to amphotericin B (>32 mg/l). 相似文献
13.
Botelho NS de Paula SB Panagio LA Pinge-Filho P Yamauchi LM Yamada-Ogatta SF 《Zoonoses and public health》2012,59(1):16-22
The aim of this study was to isolate yeasts from the faeces of urban bats inhabiting the city of Londrina, Paraná, Brazil and to determine their potential virulence attributes. Seven (12.3%) of 57 bats screened in this study showed yeasts in their faeces. Five species of the genus Candida were isolated: C. guilliermondii, C. krusei, C. lusitaniae, C. parapsilosis, and C. pelliculosa. No phospholipase activity was detected in the egg yolk plate assay; however, all isolates demonstrated protease secretion in skim milk agar. Yeasts isolated from bats produced biofilm on the surface of polystyrene plates and all were classified as intermediate biofilm producers. Minimal inhibitory concentration (MIC) values for fluconazole in the yeasts varied according to the species. Only one isolate (M34 - C. lusitaniae) was considered susceptible dose-dependent to fluconazole. The yeasts were injected intravenously into Swiss mice, and at 15 days post-infection, the animals were killed and portions of their kidneys cultured on Sabouraud dextrose agar medium. All tissues analysed showed positive cultures of Candida spp. This is the first study evaluating the presence of fungi in the faeces of bats in an urban region, where the yeast species found were shown to be potentially pathogenic. As bats are commonly found in cities, these findings indicate the need for continuous surveillance concerning environmental contamination by their excreta. 相似文献
14.
体外转录法制备新城疫病毒荧光定量RT-PCR标准阳性模板 总被引：2，自引：1，他引：1
本研究针对新城疫病毒M基因设计了1对荧光定量RT-PCR引物和Taqman探针,在上游引物的5′端引入T7启动子,采用RT-PCR方法获得了体外转录模板,并对转录产物RNA的浓度及D值进行了测定。线性试验和特异性试验结果表明,该模板具有良好的线性范围和特异性,模板稀释度为2.95×104～2.95×108拷贝/L时,相关系数为0.998。该模板稳定性好,-80℃保存30 d后无显著变化。浓度为2.95×104～2.95×108拷贝/μL的标准品的变异系数分别为0.20%、0.28%、1.00%、0.54%、0.52%,表明以此模板进行荧光定量PCR具有较好的重复性。 相似文献
15.
从辽宁某猪场采取病料,经处理后接种Marc-145细胞进行PRRSV分离,结果有2份病料在该细胞上盲传后可见典型的CPE,经测定其毒价为10-6.5TCID50/mL。间接免疫荧光试验可见黄色荧光,用电镜对纯化的病毒进行观察,可见有囊膜包裹的圆形病毒粒子,直径约为50~70 nm。该病毒对氯仿、紫外线、酸碱度变化和热敏感。对有CPE的细胞培养物进行RT-PCR,结果为阳性。接种试验动物出现典型的PRRS临床症状和死亡。结果表明,成功分离到一株PRRSV。 相似文献
16.
Micaela Sgorbini Simona Nardoni Francesca Mancianti Alessandra RotaMichele Corazza DVM 《Journal of Equine Veterinary Science》2008
The aim of this study was to evaluate whether a normal yeast flora is present in the intestine of healthy foals and whether establishing yeast flora or yeast overgrowth might be associated with foal-heat diarrhea. Among the sampled foals, 49% showed foal-heat diarrhea and 28.3% were positive for yeasts. Candida famata was isolated 9 times; Geotrichum sp., 6; Saccharomyces cerevisiae, 5; Cryptococcus laurentii, 4; Rhodotorula sp., 3; Zygosaccharomyces sp., 2; and Candida colliculosa, Candida globosa, and Candida pelliculosa 1 time, respectively. The colony-forming unit ranged from 1 to 10 in all positive cultures. Fifteen of 92 (16.3%) foals with heat diarrhea and 11 of 92 (11.9%) asymptomatic animals yielded positive samples for yeasts. None of the animals shed the same yeast more than 2 times consecutively, and yeasts were found in pure or mixed culture. Equine gastrointestinal tract did not appear to be colonized by yeasts during the first 15 days of life, and yeasts are not involved in foal-heat diarrhea. 相似文献
17.
C. J. Scott 《Equine Veterinary Education》2020,32(8):444-448
Fungal endometritis is a relatively rare cause of infertility in the mare, accounting for only 1–5% of all cases of endometritis. However, it remains a challenge to the clinician due to difficulties in diagnosing and effectively treating affected mares, resulting in a high rate of recurrence and a guarded prognosis for ongoing fertility. Often occurring as an opportunistic infection, the most common causes of fungal endometritis in the mare are yeasts (Candida spp.) and moulds with septated hyphae (Aspergillus spp.). Early detection and identification of the causative agent are vital when choosing appropriate treatment as sensitivities to commonly used antifungals vary significantly. Assessing in vitro sensitivity for each isolate is accordingly recommended. Treatment of mares with fungal endometritis is multifaceted and includes; correction of any anatomical defects, uterine lavage and systemic and/or intrauterine infusion of antifungals. Concurrent bacterial infection is common so anticipation and subsequent treatment of bacterial endometritis is also often required. 相似文献
18.
本试验利用TRIzol试剂盒提取蜜蜂球囊菌总RNA,通过Oligotex纯化试剂盒将mRNA反转录成cDNA,再以第一链cDNA为模板合成第二链cDNA,该cDNA产物经分级分离和体外包装,即获得蜜蜂球囊菌cDNA原始文库,其滴度测定为1.6×106PFU/mL,扩增后的滴度是1.5×109PFU/mL;取适量扩增文库稀释并铺平板,蓝白斑筛选独立噬菌斑,用M13±通用引物进行PCR扩增后,文库的重组率达到90%,插入cDNA片段的长度在0.5~2 kb。该文库的构建,有利于筛选目的基因,便于深入研究蜜蜂白垩病的侵染机制,最终有效防制白垩病。 相似文献
19.
为探讨牛磺鹅去氧胆酸(taurochenodeoxycholic acid,TCDCA)对热应激小鼠免疫功能的调节作用,试验将50只小鼠随机分成5组,分别为对照组、热应激组和低、中、高剂量TCDCA组(剂量分别为50、100、200 mg/kg)。热应激组和低、中、高剂量TCDCA组小鼠置于(35.5±0.5) ℃、相对湿度60%环境下连续热应激7 d。结束后分别检测小鼠脾脏指数、血清IgG、小肠sIgA的含量及外周血T淋巴细胞亚群和B淋巴细胞的变化。结果表明,热应激引起小鼠脾脏指数显著下降(P＜0.05),血清IgG、肠sIgA显著升高(P＜0.05),外周血CD8+、CD19+百分率显著下降(P＜0.05),CD4+百分率下降不显著(P＞0.05),CD4+/CD8+比值显著升高(P＜0.05),表明热应激导致小鼠机体免疫功能发生紊乱。TCDCA对热应激小鼠脾脏指数无显著影响;但能显著抑制热应激引起的血清IgG含量、小肠sIgA含量升高(P＜0.05)和显著提高外周血CD8+、CD19+百分率(P＜0.05),同时能够调节CD4+/CD8+ 比值到正常水平。因此,TCDCA对热应激导致的小鼠免疫功能紊乱具有显著的改善作用。 相似文献
20.
Alessandro Pirrone Carolina CastagnettiJole Mariella DVM PhD Cristina BonoliLaura Stancampiano DVM Maria Paola TampieriRoberta Galuppi DVM 《Journal of Equine Veterinary Science》2012
Little is known about the normal or pathologic yeast flora in healthy and critically ill neonatal foals. The aims of this study were to evaluate the yeast flora colonizing the mucous membranes of the digestive tract (oropharynx and rectum mucous membranes) of healthy and hospitalized foals and to find out risk factors involved in yeast colonization of foals referred to a neonatal intensive care unit. A total of 240 swabs were collected from 21 healthy (group A) and 39 sick (group B) foals. In 14 of the 60 foals, yeast was isolated in at least one sample (23.3%): 3 of the 21 foals (14.3%) were positive in group A and 11 of 39 foals (28.2%) were positive in group B. The yeasts were isolated from rectal swabs obtained from none in healthy foals, whereas 5 of the 39 sick foals were positive; however, this difference was not statistically significant. No significant difference was also detected regarding oropharyngeal swabs between healthy (3/21) and sick (10/39) foals. The risk factors significantly associated with the isolation of yeasts from rectal swabs were female sex, treatment with oral antibiotics, and stressful diagnostic–therapeutic procedures. The only risk factor significantly associated with the isolation of yeast from oropharyngeal swabs was the treatment with antacids and gastroprotectants. The results show that fungi present in the gastrointestinal tract of neonatal foals were mainly environmental yeasts and suggested the absence of a stable fungal colonization. Candida was the genus frequently isolated in hospitalized foals, just as it is isolated in critically ill human neonates. 相似文献