α-半乳糖苷酶在畜禽日粮中的应用研究

α-半乳糖苷是豆粕类饲料中主要的抗营养因子之一,这种产气因子复合物会降低饲料能量和蛋白质的利用率。α-半乳糖苷酶作为一种水解酶能消除豆粕饲料中的α-半乳糖苷,从而提高饲料利用率。     

大豆中α-半乳糖苷的抗营养作用及消除方法

本文综述了大豆中α-半乳糖苷的抗营养作用及α-半乳糖苷的消除方法。     

α-半乳糖苷酶的研究进展

α-半乳糖苷酶催化α-半乳糖苷键的水解,可将饲料及豆制食品中的抗营养因子α-半乳糖苷类转化分解,改善其营养成分。α-半乳糖苷酶在底物浓度富集的情况下还具有转半乳糖基作用,这一特点可用于低聚糖的合成及环糊精的改造。此外,该酶在制药、增稠剂处理和造纸工业也有一定应用。现作者对α-半乳糖苷酶的研究和应用现状,以及α-半乳糖苷酶的研究方向和应用前景进行了总结和评价。     

大豆中α-半乳糖苷的抗营养作用及消除方法

本文综述了大豆中α-半乳糖苷的抗营养作用及α-半乳糖苷的消除方法。     

α-半乳糖苷酶在畜禽中的研究进展北大核心

α-半乳糖苷酶是催化水解α-半乳糖苷末端半乳糖残基的酶类,能够催化棉籽糖、水苏糖水解等,在自然界中广泛存在。α-半乳糖苷可以分解饲料原料中的抗营养因子,显著提高畜禽对豆粕以及豆类原料的养分利用率,改善畜禽生产性能。文章就α-半乳糖苷酶的结构、酶学特性、其作用机制以及在畜牧业中的应用进行阐述,为畜禽饲料添加剂的开发提供参考。     

α-半乳糖苷酶对猪生长性能及营养物质体外消化率的影响

试验主要研究日粮中添加不同水平的α-半乳糖苷酶对猪生产性能、死亡率和营养物质表观消化率的影响。试验选取健康、体重均匀、27日龄的断奶仔猪192头,随机分为4个处理,每个处理6个重复,每重复8头,公母各半。日粮α-半乳糖苷酶添加量为50 U/kg、100 U/kg和200 U/kg,试验期为150 d。结果显示:日粮中添加α-半乳糖苷酶,能显著降低仔猪腹泻率和死亡率(P 0.05),在育肥和全期阶段,α-半乳糖苷酶可显著降低料重比(P 0.05);显著提高粗蛋白和粗纤维的表观消化率(P 0.05)。结果提示:日粮中添加α-半乳糖苷酶,可显著提高营养物质的消化率,降低仔猪腹泻率,添加量为100 U/kg时效果最好。试验旨在研究不同α-半乳糖苷酶添加量对猪生产性能和主要营养物质表观消化率的影响,为进一步推广α-半乳糖苷酶制剂在猪日粮中的应用提供数据支撑。     

α-半乳糖苷酶的性质研究

文章主要测定了α-半乳糖苷酶酶活、pH对酶活的影响,以及α-半乳糖苷酶的耐热性、耐酸性。结果表明,α-半乳糖苷酶各方面的性质都较优越,作为饲料添加剂,能起到降解饲料中存在的半乳糖苷等抗营养因子,促进蛋白质的合成,提高瘦肉率;降低豆类原料引发的幼龄动物腹泻;降低动物维持需要量,提高饲料能量效价;减少饲料中抗生素的用量等作用。     

复合酶中补充α-半乳糖苷酶对蛋鸡生产性能的影响

试验选择25周龄健康海兰灰粉蛋鸡5000只,按单因素设计随机分成2个处理,每处理5个重复,每重复500只。分别饲喂对照日粮和添加100ml/tα-半乳糖苷酶的试验日粮,目的是研究在添加FE808E蛋禽复合酶的蛋鸡日粮中补充α-半乳糖苷酶对蛋鸡产蛋性能和经济效益的影响。结果表明:在整个试验期内蛋鸡日粮中补充α-半乳糖苷酶,通过配方调整可降低蛋鸡饲料成本42元/t,提高蛋鸡的产蛋率1.24%、增加蛋重0.77%、降低料蛋比0.97%、并使每千克蛋的生产成本降低0.13元,同时能显著降低脏蛋率(P0.05)。表明在FE808E复合酶基础上补充α-半乳糖苷酶能进一步提升FE808E蛋禽复合酶产品的功能和提高蛋鸡生产性能及饲料转化率。     

产α-半乳糖苷酶菌株的筛选、鉴定及其酶学特性的研究

利用筛选培养基从不同豆制品作坊附近的土样中筛选出一株产α-半乳糖苷酶的菌株D-1,对其进行分子鉴定及所产的α-半乳糖苷酶进行酶学特性研究。研究表明,菌株D-1为Aspergillus niger,此菌株所产α-半乳糖苷酶的最适反应温度是55℃,在60℃以下热稳定性较好;最适反应pH值为5.0,在pH值3.0～5.5范围内稳定性较好,相对酶活>64.1%;Mg2+、Na+、Pb2+、K+、Mn2+、Co2+、Al3+对α-半乳糖苷酶均有不同程度的抑制作用,其中Cu2+和Fe3+的抑制作用较为明显,而Ca2+、EDTA(乙二胺四乙酸)、Zn2+对α-半乳糖苷酶有一定的促进作用。     

α-半乳糖苷酶和水苏糖对断奶仔猪生长性能的影响

研究选用72头体重(7.74±0.23)kg断奶仔猪,随机分成4组,按2因素方差设计,即2个水平水苏糖(1.26%和2.16%)和2个水平的α-半乳糖苷酶(0%和0.01%)。试验处理如下:①基础日粮;②基础日粮加酶;③基础日粮加水苏糖;④基础日粮加水苏糖加酶。研究表明,高水平的水苏糖可显著性地降低断奶仔猪生长性能(P<0.05))。日粮中添加α-半乳糖苷酶可以提高断奶仔猪的生长性能(P<0.05));α-半乳糖苷酶可以显著性的提高日粮中水苏糖、棉籽糖、总能和粗蛋白的消化率;仔猪消化道的微生物区系受日粮中水苏糖的水平和α-半乳糖苷酶的影响。     

Generation of Sperms Containing EGFP‐LacZ Following Transfection of Chicken Testis with a Eukaryotic Dual Reporter Vector

Male germ cells modified by foreign genes can be used to generate transgenic chicken. In this study, in vivo transfection of chicken testis with an EGFP‐LacZ dual reporter expression vector was performed. Large‐scale plasmid DNA preparation of the EGFP‐LacZ eukaryotic expression vector was carried out and efficient transfection of chicken testicle cells using the prepared plasmid DNA was confirmed in vitro. The reporter plasmid was directly injected into adult rooster testes. Semen samples were collected on 10‐days post‐transfection and every other day thereafter; and a total of six collections were made. Semen slides were subjected to fluorescence microscopy and β‐galactosidase activity assay to identify sperms carrying the reporter genes. The presence of EGFP and LacZ was further confirmed by PCR amplification with sperm genomic DNA as template. The testicles of those birds were subjected to cryostat sectioning, fluorescence microscopy and β‐galactosidase activity assay. The results showed that sperms with green fluorescence were not observed on semen slides; however, sperms positive for β‐galactosidase were detected. Specific amplicons of EGFP and lacZ were detected in four of the six sequentially collected semen samples. Fluorescence microscopy of the corresponding semen slides revealed yellow‐green fluorescence, but not clear green fluorescence. The β‐galactosidase activity assay and GFP histochemistry using monoclonal antibodies demonstrated positive staining for subsets of testicle cells. Together, these results showed that direct injection of the dual reporter vector into adult rooster testis allowed in vivo transfection of chicken sperm precursor cells, which further developed into sperm containing EGFP‐LacZ.     

Follicle dynamics and characteristics of ovulation in heifers after Ovsynch treatment in the last third of the estrous cycle

The objective of the experiment was to study follicular dynamics and characteristics of ovulations in dairy heifers after application of the Ovsynch protocol in the last third of estrous cycle. Therefore, altogether 27 regular cycling Holstein heifers were given an injection of GnRH on day 14, 16 or 18 (9 heifers each in group 1 to 3) of the estrous cycle. All heifers were administered PGF2alpha seven days later. Blood was collected for progesterone determination, just before, 24 hours and 48 hours after the PGF2alpha injection. A second injection of GnRH was administered 48 hours after the PGF2alpha injection. Ovarian follicular dynamics were monitored by frequent ultrasound scanning of the ovaries after first and second GnRH injection. Altogether 22 of 27 heifers (81.5%) ovulated 27 to 33 h after first GnRH injection. In 4 heifers ovulations were recorded 45 to 51 h after first GnRH application. Mean intervals between GnRH application and ovulation were 33.0, 33.6 and 28.3 h, respectively. At the time of PGF2alpha injection mean progesterone concentrations were similar in groups 1 and 2, but significantly lower than in group 3. After the second GnRH treatment 5,6 and 8 heifers had ovulations.The average intervals from the second GnRH treatment to ovulation were 24.8, 24.0 and 24.4 h respectively.The results show that Ovsynch is not sufficient to ensure synchronisation of oestrous and ovulation in each animal treated.     

Effects of dietary addition of Macleaya cordata alkaloid extract on growth performance,caecal indices and breast meat fatty acids profile in male broilers

The aim of this experiment was to investigate whether a low‐dietary application (15 mg/kg) of an alkaloid preparation containing quaternary benzo[c]phenanthridine alkaloids and obtained from Macleaya cordata (Sangrovit) influenced caecal metabolism, growth performance and long‐chain fatty acid composition of breast meat. One‐day‐old broiler chicks (Cobb 500) were fed a diet without supplement or with a 15 mg/kg dose of Sangrovit (C and A groups respectively) for 5 weeks. Although the A treatment was not accompanied by an enhanced final body weight of broilers, the intake of a diet with Sangrovit influenced the caecal microflora activity. The addition of Sangrovit to a diet decreased potentially harmful β‐glucuronidase and β‐glucosidase activities (p < 0.05 and p = 0.075 respectively), and at the same time led to a significant increase in activities of bacterial glycolytic enzymes α‐glucosidase, α‐galactosidase, β‐galactosidase in comparison to the control group. The concentration of total short‐chain fatty acids in the caecal digesta was increased in the A treatment contributing to the tendency towards lower caecal pH (p = 0.078). The analysis of breast meat fatty acids showed that the dietary application of Sangrovit evoked some changes in contents of monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) contents. The applied dosage of Sangrovit caused an increase in the sum of MUFA and the tendency towards lower PUFA sum (p < 0.05 and p = 0.062 respectively) as in relation with the C group. Although the nutritionally relevant n‐6/n‐3 PUFA and the (PUFA + MUFA)/saturated fatty acids ratios remained similar in both groups, further research is postulated to establish the effect of this preparation on meat quality. To sum up, despite of a lack of the improvement in final body weight, a low dose of dietary Sangrovit was found to exert positive effects on the caecal metabolism of the broilers.     

Estrus and ovulation synchronization in Merino meat sheep. 1. Effect of Gonavet "Berlin Chemie" after estrus synchronization with prostaglandin F2 alpha in Merino meat sheep]

Oestrus synchronisation by means of PGF2 alpha analogues was followed by injection of Gonavet "Berlin-Chemie" which triggered off an LH peak, 2 to 3 hours from injection. Injection of Gonavet "Berlin-Chemie", 44 hours after PGF2 alpha application, caused synchronisation of all LH peaks. The interval between injection of Gonavet "Berlin-Chemie" and onset of ovulation amounted to 22 hours. The length of ovulation was not accurately determinable. Ovulation was successfully induced to all sheep by application of Gonavet "Berlin-Chemie", 44 or 48 hours after PGF2 alpha injection. Ovulation rates were 1.75 or 1.54. Luteolytic action on sheep of Cloprostenol "Jenapharm", a PGF2 alpha analogue, proved to be just as good as that of Oestrophan (SPOFA).     

Preventive effect of hyaluronic acid on the suppression of attachment and migration abilities of bovine chondrocytes by IL-1alpha in vitro

Attachment and migration of bovine chondrocytes cultured in vitro were significantly suppressed by the addition of interleukin (IL)-1alpha at the concentration of 1 ng/ml or more (p<0.05). The application of hyaluronic acid (HA) at the concentration of 10 micro g/m l or more significantly recovered the attachment of chondrocytes (p<0.05) and the application of HA at 100 micro g/ml concentration recovered the migration of chondrocytes suppressed by IL-1alpha. These results suggest that the application of HA for inflammatory arthropathies or chondrocyte transplantation might be helpful to preserve the properties of chondrocytes and its extracellular matrix against inflammatory conditions.     

驯鹿干扰素β1基因的克隆及遗传进化分析

试验旨在从驯鹿鹿茸顶端组织中克隆干扰素β1（interferon beta 1,IFNβ1）基因全长序列,分析其分子特性,为研究其生物学活性及实际应用奠定基础。根据GenBank数据库中牛、羊等近缘物种IFNβ1基因的保守序列设计1对引物,以驯鹿鹿茸顶端间充质层组织基因组DNA为模板,采用PCR技术扩增得到驯鹿IFNβ1基因并克隆、测序,利用相关生物信息学软件对该序列进行分析。结果发现,驯鹿IFNβ1基因全长561 bp,共编码186个氨基酸,含有3个糖基化位点;其编码蛋白含有4个α螺旋、4个β折叠区、7个β转角。将驯鹿IFNβ1基因推导的氨基酸序列与其他14种哺乳动物进行遗传进化分析发现,其同源性为45.1%~92.0%;驯鹿与其他动物IFNβ1基因序列分析和系统进化树分析表明,IFNβ1基因存在种属差异性,亲缘关系越近,同源性越高。本研究结果为驯鹿IFNβ1基因的进一步研究和应用提供了基础试验数据。     

土槿皮乙酸B增加活性氧水平诱导人乳腺癌MCF-7细胞凋亡和衰老的研究

本研究旨在研究土槿皮乙酸B(PAB)诱导人乳腺癌MCF-7细胞凋亡和衰老与活性氧的关系。在相差显微镜下观察4 μmol/L PAB在36 h诱导细胞凋亡小体的出现;LDH方法显示PAB时间依赖性的增加凋亡比率;SA-β-半乳糖苷酶显示4 μmol/L PAB作用3 d后,去药再作用5 d,96%的细胞蓝染,细胞变大且扁平;流式细胞仪检测用DCFH-DA染色后4 μmol/L PAB 时间依赖性增加细胞内活性氧水平。所以4 μmol/L PAB通过增加细胞内活性氧水平促进细胞凋亡和衰老。     

In vitro safety assessments and antimicrobial activities of Lactobacillus rhamnosus strains isolated from a fermented mare's milk

Safety and probiotic characteristics such as antimicrobial activities of three Lactobacillus rhamnosus strains, FSMM15, FSMM22 and FSMM26, previously isolated as potential probiotics from fermented mare's milk were investigated. The three FSMM strains were susceptible to ampicillin, gentamycin, kanamycin, streptomycin, tetracycline and chloramphenicol, whereas they were resistant to erythromycin (minimal inhibitory concentration (MIC) = 4?8 µg/mL) and clindamycin (MIC = 4 µg/mL); bioconversion of bile salts, hemolytic activity and mucin degradation activity were negative; enzymatic activities of α‐chymotrypsin and β‐glucosidase were detected, but those of α‐galactosidase, β‐glucuronidase and N‐acetyl‐β‐glucosaminidase, were undetectable. Among the strains, strain FSMM15 was chosen as a safer probiotic candidate due mainly to the lack of plasminogen binding ability. Despite lower acid production of strain FSMM15 than others, its cell‐free culture supernatant inhibited growths of Salmonella Typhimurium LT‐2, Shigella sonnei , Listeria monocytogenes , and Escherichia coli O157 with comparable levels of ampicillin, suggesting a favorable aspect of strain FSMM15 as a probiotic strain.     

Characterization of alpha-adrenoceptor subtypes in smooth muscle of equine ileum

OBJECTIVE: To determine the concentration and binding characteristics of alpha-adrenoceptor subtypes in smooth muscle cell membranes of equine ileum. SAMPLE POPULATION: Segments of longitudinal and circular smooth muscle from the ileum of 8 male and 8 female adult horses. PROCEDURE: Distribution of alpha-adrenoceptor subtypes was assessed by use of radioligand binding assays incorporating [3H]-prazosin and [3H]-rauwolscine, highly selective alpha1- and alpha2-adrenoceptor antagonists, respectively. Characterization of adrenoceptor subtypes was performed by use of binding inhibition assays. RESULTS: On the basis of binding affinity for specific radioligands, low- and high-affinity alpha1- and alpha2-adrenoceptors were detected. Concentration of low-affinity alpha2-adrenoceptors was significantly greater in male horses, compared with females. Competition studies confirmed the specificity of the radioligands used in the binding assays. Alpha1-adrenoceptors of both subtypes in male and female horses had a higher affinity for prazosin than phentolamine, whereas yohimbine did not compete with the radioligand for binding. For alpha2-adrenoceptors regardless of subtype, potency of inhibition elicited by each drug varied between sexes. In males, yohimbine was a more potent inhibitor than phentolamine, which was more potent than prazosin. In females, yohimbine was more potent than prazosin, which was more potent than phentolamine. CONCLUSIONS AND CLINICAL RELEVANCE: High- and low-affinity alpha1- and alpha2-adrenoceptors were detected in smooth muscle of equine ileum. Because alpha-adrenoceptor subtypes, particularly alpha2-adrenoceptors, are involved in the regulation of gastrointestinal tract function, characterization of these receptors may represent the basis for development of new therapeutic strategies for the control of gastrointestinal disturbances in horses.