初乳中乳铁蛋白的分离纯化及鉴定

乳铁蛋白(Lactoferrin,Lf)是一种具有许多独特生理功能的铁结合糖蛋白,主要存在于哺乳动物的各种外分泌物中,如乳汁、眼泪等,乳汁尤其是初乳中Lf的含量很高。因此,从来源丰富的牛初乳中提取Lf,把它作为功能性成分应用于食品和畜牧生产中,具有广阔的前景。本试验通过等电沉淀、盐析和层析分离等方法从牛初乳中分离Lf,并用SDS-PAGE、Western Blot检测和鉴定,结果我们得到了80 ku的乳铁蛋白,其纯度90%,符合工业化生产要求。本试验建立的方法简便、经济、安全,可用于乳铁蛋白的工业化生产。     

牛初乳中乳铁蛋白粗提工艺的优化研究

利用盐析和有机溶剂沉淀相结合的方法对牛初乳中的乳铁蛋白进行初步的分离,采用单因素试验和正交试验的方法对影响分离效果的因素进行了试验研究,研究结果表明:用pH值8.5、80%的硫酸铵溶液进行盐析,然后用60%的乙醇浓度进行沉淀,分离的效果较好。SDS-PAGE凝胶电泳结果表明,乳铁蛋白的纯度大约为48%,分子量大约为78000Da。     

牛初乳中生长因子及抗菌因子的生物活性及其功能研究进展

主要介绍了牛初乳中富含生理活性功能的活性成分——生长因子(胰岛素样生长因子、转化生长因子、表皮生长因子、成纤维细胞生长因子)及抗菌因子(乳铁蛋白、溶菌酶、乳过氧化物酶、免疫球蛋白)生理活性和功能的最新研究进展,同时,对牛初乳中各因子作为保健产品和医疗药品基料的趋势作出展望。     

婴幼儿配方羊奶粉中乳铁蛋白含量测定

以食品安全国家标准《食品中乳铁蛋白的测定(征求意见稿)》为参考,优化预处理步骤和梯度洗脱条件,肝素亲和柱净化,紫外高效液相色谱法测定婴幼儿配方羊奶粉中乳铁蛋白含量。检测波长280 nm,乳铁蛋白标准品出峰时间9.475min。乳铁蛋白标准品工作液浓度50~250μg/mL,在此范围内线性回归方程y=129.67x-7.4719,R~20.9991。乳铁蛋白定性重复性RSD%(n=6)为0.074,定量重复性RSD%(n=6)为0.996。乳铁蛋白加标回收率在93.4%~99.1%,平均回收率96.7%。     

特殊医学用途婴儿配方食品中乳铁蛋白的检测

建立特殊医学用途婴儿配方食品中乳铁蛋白的高效液相色谱（high performance liquid chromatography,HPLC）检测方法。试样中的乳铁蛋白经磷酸盐缓冲液提取,肝素亲和柱净化后,用反相-HPLC分离,以0.1%三氟乙酸和乙腈作为流动相进行梯度洗脱,HPLC仪紫外检测器检测,外标法定量。结果表明：在乳铁蛋白质量浓度50～500 μg/mL范围内,检测结果呈现良好线性关系;该方法的检出限为5 mg/100 g,定量限为15 mg/100 g,平均回收率为93.9%;同一质量浓度乳铁     

我国牛初乳资源及其安全性评价

牛初乳总固体含量高,富含免疫球蛋白、乳铁蛋白等生物活性因子,具有较高的营养保健作用。我国1～3 天牛初乳年产量约20.2 万吨,除哺喂犊牛外,约1/2可供加工利用。研究表明,牛初乳对人体具有调节免疫、促进生长发育、增进肠道健康、延缓衰老等保健作用。我国牛初乳资源丰富,发展潜力大,应规范牛初乳制品市场,制定食品安全国家标准,加强科普宣传和质量监管,促进牛初乳产业健康、有序发展。     

高效液相色谱法测定乳品中乳铁蛋白的含量

建立乳品中乳铁蛋白高效液相色谱的测定方法。采用pH值为7.0磷酸盐缓冲溶液做为提取剂,C_(18)色谱柱分离,不同浓度的乙腈-0.5 mol/L的氯化钠溶液-三氟乙酸的二元系统进行梯度洗脱。每100g奶粉(或每100mL液态奶)中乳铁蛋白添加量为5~10 mg时,回收率大于90%。定量限为0.5 mg/100g。该方法具有结果准确,操作简便,重复性好等优点,可用于乳品中乳铁蛋白的检测。     

牛初乳中乳铁蛋白的分离纯化及其抗菌活性研究

本试验以饱和(NH4)2SO4盐析法从江西南昌市某牛场母牛初乳中分离到乳铁蛋白(lactoferrin,LF),经Sephadex G-100葡聚糖凝胶过柱纯化,并利用聚乙二醇(PEG6000)浓缩获得了牛初乳乳铁蛋白.同时用猪常见细菌病病原猪大肠杆菌(E.coli.)、猪传染性接触性胸膜肺炎放线菌(APP)以液体培养...     

天然多功能性原料乳铁蛋白无菌液关键技术的开发

乳铁蛋白属于热敏性物质，在经过超高温瞬时（120℃，4 s）处理后乳铁蛋白变性率高达91.10%。本文通过对乳铁蛋白原料的溶解度、粘度、稳定性等不同特性的研究及膜过滤的研究，成功研制出液态的无菌乳铁蛋白原料，并可以投入批量生产。该产品实现了世界首次无菌在线添加。     

反相高效液相色谱法检测牛初乳乳铁蛋白的方法研究

为了建立反相高效液相色谱法(RP-HPLC)测定牛初乳乳铁蛋白的含量,采用Kromasil C4色谱柱(250 mm×4.6 mm,5μm),流动相:乙腈-水-三氟乙酸(80:20:0.1)流速0.6 mL/min;检测波长:220nm;柱温:25℃.乳铁蛋白的检测线性范围为0.025-1 mg/mL,回归方程为Y=734105X-8035.9,相关系数r=0.999,平均回收率为99.58%,相对标准偏差为1.3%.此方法操作方便、快捷、精密度好、结果准确可靠、重复性好.     

Effect of cow colostrum on the performance and survival rate of local newborn piglets in Benin Republic

The effect of bovine colostrum, including its thermally labile compounds, on the survival and growth performance of local breed piglets reared by their mother, in Benin, was evaluated over a 49-day trial. Three groups of 16 piglets, stemming from two primiparous sows belonging to a unique traditional farm, were respectively fed for the first 48 h of life with either bovine colostrum heated to 85 °C for 30 min, or thawed bovine colostrum, or colostrum from the mother. Thereafter, the animals that received bovine colostrum turned back to their mother. At day 21, almost all piglets from the group that received heated colostrum died. The highest total weight gain was obtained in the group that received thawed bovine colostrum (P???0.01), followed by the group left with the mother. Corresponding average daily gains (ADGs) were 56, 34 and 2 g/day, respectively (P???0.05). At the end of the trial, the treatment effect was highly significant on the survival of piglets (100% in the thawed colostrum group vs. 00 and 50%, respectively, in the heated colostrum group and in the group left with the mother). At day 49, numerically higher weight and ADGs were obtained in the group that received thawed cow colostrum. Thawed bovine colostrum improved the growth performance and piglet survival in the local pig breed in Benin, probably owing to thermally labile components. Bovine colostrum may be used in our farms in order to reduce pre-weaning mortality, improve the profitability of livestock farmers, and ensure survival of traditional farms. The use of bovine colostrum on farms could be facilitated by collaboration between pig farmers and bovine farmers. It could also be facilitated by the creation of a colostrum bank.     

Composition and physiological functions of the porcine colostrum

The first secretion, 24-h post parturition of the mammary glands of sows, known as colostrum, is high in protein and low in lactose and fat. As a consequence of an insufficient ingestion of colostrum, more than 50% of piglets fail to reach weaning and die. The composition and some functions of colostrum have been previously reported. For example, colostrum carbohydrates consist of mainly lactose. Lipids in the colostrum are mostly triacylglycerols, but <1% is fatty acids, which may act as homeostasis regulators. Similarly, proteins are found mostly as casein and whey, the latter being ≥80% immunoglobulins. Colostrum-derived immunoglobulins and bioactive proteins such as azurocidin help the immune system of the piglet fend off infections. In addition, leukocytes and exosomes are other minor but nonetheless equally crucial bioactive components in the porcine colostrum. Modern pig farming has achieved increases in pig productivity and litter size, but this has been accomplished in detriment of the health and the survival rate of piglets. Therefore, porcine colostrum is now even more important in pig farming. In the present review, we discuss the current knowledge on the composition and physiological functions of the porcine colostrum and briefly propose future research directions.     

Investigation of some factors which influence the absorption of IgG by the neonatal piglet

The absorption by the neonatal gut of purified swine IgG isolated from maternal serum was studied in 91 motherless piglets raised without swine colostrum, to investigate some of the factors which influence absorption. Factors studied included the amount of IgG administered, the effect of bovine colostrum fed concurrently, the influence of starvation and the effect of lactose.Administration of iodinated IgG within 3 hours after birth resulted in the appearance of IgG in the piglet circulation in unaltered form. Immediate feeding of bovine colostrum followed by administration of iodinated IgG at 72 hours resulted in the appearance of only IgG digestion fragments in the circulation.Administration of 15 g of lactose during the first 24 hours reduced absorption of IgG by 26% compared to controls while treatment with 54 g of lactose reduced absorption by 94%. On the contrary, when bovine colostrum was administered immediately after purified IgG, the amount of swine IgG absorbed was 50–70% greater than in controls. Mature milk failed to have the same influence. Finally, in experiments in which the amount of purified swine IgG administered varied from 1.0 to 8.0 g, absorption was directly proportional to the amount administered. When these data were compared with data on naturally reared piglets, the same proportionality was seen and indicated a consistent 1:2.5 to 1:3 ratio of ingested IgG (g) to the maximal serum level of immunglobulin (mg/ml) resulting from absorption. IgA and IgM follow a similar pattern.The data are consistent with a regulated absorption mechanism, that appears to be lost very quickly after the initial ingestion of food. Heterologous protein and lactose alone can switch off this absorption mechanism, thus arguing against “gut closure” by a mechanism of receptor saturation by absorbed IgG. Bovine colostrum, when compared to mature milk, is able to augment the absorption of swine IgG by an unknown mechanism.     

Relative contribution of colostrum from Maedi-Visna virus (MVV) infected ewes to MVV-seroprevalence in lambs

Maedi-visna virus (MVV) seroprevalence associated with consumption of colostrum from seropositive ewes was investigated in 276 housed lambs from birth to 300 days-old. At birth, lambs were allocated to five experimental groups according to the maternal MVV-serological status, source and mode of feeding colostrum (bovine or ovine and bottle fed or suckled from the dam) and type of horizontal MVV-exposure (raised with the dam or separately with other lambs). The risk of being seropositive at 300 days-old was associated with feeding ovine colostrum from seropositive ewes and increased with intake of bottle-fed ovine colostrum and was higher in lambs separated from their dams and raised with other experimental lambs compared to lambs raised with their dams. Approximately 75-87% of ELISA-positive results in lambs that had ovine colostrum was attributable to colostrum itself. However, approximately only 16% of naturally raised and 29-61% of bottle-fed ovine colostrum lambs were ELISA-positive as a result feeding ovine colostrum. These results confirm that ovine colostrum from seropositive ewes can be a major source of MVV but its overall contribution to seroprevalence in natural conditions is relatively low, and shows that horizontal MVV transmission can be an important source of infection in new-born lambs.     

Studies on the absorption of homologous and heterologous IgG in artificially reared newborn pigs

The interaction of homologous and heterologous IgG during intestinal absorption was investigated using five groups of newborn piglets (50 animals in total). The diet, given via a stomach tube, was different in each group during the first 24 h. Group I received bovine colostrum, group II bovine colostrum and porcine IgG solution, group III bovine and porcine colostrum, group IV bovine colostrum and intraperitoneally applied monomeric porcine IgG, and group V received a glucose diet with no immunoglobulins. Feeding was based on bovine colostrum between the 2nd and 6th days after birth, followed by a milk replacer during the rest of the experimental period. The serum concentrations of homologous and heterologous IgG were monitored from birth to 10 weeks of age. The total serum IgG content (homologous + heterologous) of newborns was almost equal in groups I–IV at 24 h. Porcine IgG from endogenous synthesis was detectable in the serum of groups I and V two weeks postpartum. The heterologous IgG absorbed from bovine colostrum produced nearly the same serum concentration in groups II and III: hence the different components of porcine colostrum did not influence the absorption of heterologous IgG. Intraperitoneal application of 1.3 g porcine IgG in group IV resulted in a delay of the synthesis of endogenous IgG. The average half-life of heterologous IgG in the serum of groups I–IV was almost exactly the same, showing that the porcine colostrum or IgG solution did not modify the half-life of bovine IgG. The ingestion of the glucose diet within the first 24 h completely blocked the absorption of IgG from bovine colostrum applied from the second day. Possible explanations of the phenomena investigated are discussed.     

Maternal-neonatal immunoregulation: suppression of de novo synthesis of IgG and IgA, but not IgM, in neonatal pigs by bovine colostrum, is lost upon storage

Fifty-four neonatal pigs were allotted to 4 groups and reared in an electrically controlled automatic feeding device (autosow). Each group was reared on a different pool of bovine colostrum: fresh, stored 1 month, stored 6 months, and stored 8 years. Bovine and porcine immunoglobulins in the sera of these pigs, and in a group of conventionally reared pigs, were measured periodically during the first 42 days after birth. The maximal concentration of absorbed bovine immunoglobulin was reached between 12 and 18 hours and equaled or exceeded the amount of porcine immunoglobulin absorbed by the conventionally reared pigs. Large differences in the concentrations of the bovine immunoglobulin isotypes among the various pools of colostrum were positively correlated with concentration of these isotypes in the sera of the neonatal pigs fed these pools. Relative to their concentrations in colostrum, approximately 41% of the IgG1, 55% of the IgG2, 29% of the IgM, and 67% of the IgA was absorbed. The IgA was absorbed the best and IgM was least absorbed. Significant trends or differences in absorption were not observed among groups. Neonatal pigs given fresh colostrum, which had a higher fat content, had significantly more weight gain (P less than 0.05). This occurred, despite the fact that the fresh colostrum had the lowest concentration of bovine immunoglobulin. Serum half-lives for bovine IgG1 and IgG2 were significantly less than for porcine IgG (P less than 0.05), whereas the half-lives for bovine and porcine IgM and IgA were similar. De novo-synthesized immunoglobulins were detectable in serum after 6 days; IgM concentrations reached a maximum at 15 days in neonatal pigs given stored, but not fresh, colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

The absorption of colostral immunoglobulins in newborn piglets. III. Influence of the duration of colostrum administration

The gastrointestinal tract of newborn piglets is permeable for intact immunoglobulins ingested with the colostrum. The duration of this passage was investigated by administering hourly rations of 25 ml of either porcine or bovine colostrum for 6, 12, 18 or 24 hrs after birth. The plasma concentrations of the subclasses porcine IgG, IgM and IgA or bovine IgG1, IgG2, IgM and IgA were determined at 12, 18 and 24 hrs after birth and on days 3 and 6. Feeding periods of 6 hrs resulted in plasma Ig levels of the same order of magnitude as observed in natural rearing. These levels were not substantially increased after prolonged feeding. The 6% gain from 6 to 12 feedings seen with porcine colostrum as compared with a gain of 24% for bovine colostrum points at an earlier closure of the intestinal wall for the species-specific proteins. There was no further increase of Ig permeation after 12 hourly feedings. Growth performances and losses were identical in all groups.     

The effect of dietary spray-dried bovine colostrum and plasma on the response of pigs to enterotoxigenic E. coli challenge after weaning

Twenty four, 21-d-old female pigs were fed diets containing either skim milk powder (CON+), spray-dried bovine colostrum (7.5%, BC+) or spray-dried bovine plasma (7.5%, BP+) and were dosed orally with 1 × 10⁹ CFU of E. coli O149:K88. Another group of 8 unchallenged pigs was fed the skim milk powder diet (CON−). On d 19 of the experiment all piglets were euthanased. Adverse effects of the E. coli challenge were observed variously throughout the small intestine in pigs consuming either the BC+ or BP+ diets. In this experiment, similar responses to the E. coli challenge were observed in both plasma and colostrum fed pigs, which suggests that spray-dried bovine colostrum may be a potential alternative to spray-dried bovine plasma.     

Antioxidants in Bovine Colostrum

Quality, content and properties of colostrum are crucial for the neonate and its further development. Due to essential differences between intrauterine and extrauterine environment, the neonate is exposed to oxidative stress conditions. Colostrum apart from nutrient and immunological components should contain antioxidative systems necessary for the protection against reactive oxygen species. This review describes available data on enzymatic and non-enzymatic antioxidants in colostrum. Due to the fact that the literature concerning bovine colostrum is scanty, the information based on bovine mature milk determinations as well as other species is provided. Bovine colostrum is used not only by calves, but also for the production of hyperimmunized colostrum, medicines or feed supplements. Quality of colostrum influences quality of mature milk. This is another reason, except from health of neonate, why antioxidative properties of bovine colostrum are of special importance and require further detailed elucidation.