Colostrogenesis during an induced lactation in dairy cattle

Colostrum immunoglobulin G (IgG) is of major importance for the newborn calf because epitheliochorial placentae do not provide transport in utero. The formation of colostrum occurs in the later stages of pregnancy. Our objectives were to induce lactation in non‐pregnant dairy cows and (i) to determine the changes of IgG in serum and mammary secretions during the induction process and (ii) to establish α‐lactalbumin (αLA) and prolactin (Prl) alterations to monitor the changing mammary epithelial tight junction status and development pattern. Estradiol‐17β (E2) and progesterone (P4) injections in a 1–7 days series were combined with a 3‐day injection series (day 21–23) of dexamethasone (DEX). Blood and both front quarter secretion samples were collected daily. Milking started 24 days after the start of the experiment. Results show that the mammary secretory IgG1 was increased at >7 days after the start of steroid injections and depicted a bimodal pattern reaching a high of 16 mg/ml at 21 day compared with 3.2 mg/ml in the serum. There was a small increase in secretory IgG2 that did not correlate with tight junction status, but never reached the serum concentration. The injections of DEX resulted in constriction of tight junctions. Secretory αLA was immediately increased with steroid injections, dropped precipitously after 7 days and then began a steady increase until the start of milking. Changes in serum αLA are related to mammary tight junctions while serum Prl gradually increased from 30 to >60 ng/ml after the steroid injections stopped. These results provide insights into the mechanisms and timing of colostrogenesis during an induced lactation protocol.     

微波处理条件对蛋清结构和IgG结合能力的影响

为探究降低蛋清致敏性的新方法,在不同的微波条件下处理蛋清蛋白,通过SDS-PAGE(十二烷基磺酸钠-聚丙烯酰氨凝胶电泳)分析分子量变化、圆二色光谱分析二级结构变化、紫外光谱分析三级结构变化、外源性荧光光谱分析疏水性变化、间接ELISA(酶联免疫吸附试验)分析IgG(免疫球蛋白G)结合能力的变化,研究微波对蛋清的影响。研究证实,微波处理降低蛋清蛋白的潜在致敏性,其中400 W/30 s处理后的蛋白显示出最低的IgG结合能力,所有条件下处理后的蛋清蛋白的二、三级结构均发生显著改变。80 W的微波功率下,蛋白质的结构先折叠,该条件下处理30 s后,蛋白结构又展开;当微波功率达到640 W时,处理时间较长的蛋白分子产生堆叠。蛋白质分子的结构变化导致了其抗原性的变化。     

Evaluation of Brix Refractometry for the Estimation of Colostrum Quality in Jennies

Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right half of nine Amiata jennies. A total of 72 colostrum samples were analyzed. A Friedman test with a Dunn’s test for multiple comparisons was used for assessing the differences between the left and right half at each sampling time. No differences were found between the left and right halves; the average value was used to analyze the effect of sampling time on the IgG concentrations and Brix values. The relationship between colostrum IgG concentrations (SRID test) versus Brix value and Brix value versus time was analyzed using two different mixed linear models. A strong statistically significant relation has been found between IgG concentrations and Brix values (R² = 0.84). The relation between IgG concentrations and Brix refractometer showed a cutoff point of 17% Brix score for the identification of high-quality colostrum. The Brix value (%) decreased continuously from 16.29 by 0.29 × hour. Jennies’ and donkey foals’ management may be greatly improved using this simple and cheap device.     

兔抗猪瘟高免血清的制备及纯度鉴定

利用猪瘟弱毒疫苗(C株)免疫新西兰大白兔,制备抗猪瘟高免血清IgG。运用间接ELISA方法测血清效价,再经饱和硫酸铵盐析沉淀法提纯血清IgG,SDS-PAGE电泳法鉴定提纯IgG的纯度,结合无菌检查试验和仔猪接种试验,检测血清IgG的安全性。结果表明,获得了蛋白含量为6 mg/mL,效价为1∶6400的高效价、安全性高、成本低的抗猪瘟高免血清IgG。为猪瘟病毒的检测和猪瘟的防制提供了理想的生物制剂。     

猪流行性腹泻病毒血清IgG间接ELISA检测方法的建立

以大肠杆菌表达系统制备的猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)中和抗原S1D(1μg/mL)每孔100μL进行包被,利用HRP-兔抗猪IgG(1∶20 000)建立了检测猪血清中抗PEDV IgG的间接ELISA方法。阴性判断临界值(M+2SD)为0.632,批内和批间检测的平均变异系数分别为2.3%和3.1%,该方法与商品化的PEDV抗体间接ELISA检测试剂盒同时检测50份临床血清样品,均为阳性,上述结果表明该方法可用于PEDV血清IgG的临床检测。     

初乳清粉的货架期预测和贮藏稳定性

为了掌握初乳清粉的贮藏特性,采用货架期预测方程来估计初乳清粉在室温(25℃)、50％相对湿度(RH)条件下PET包装的货架期,并对90d贮藏期过程中其理化指标以及生化指标的变化进行研究.研究结果表明:在PET包装90d贮藏时间内,初乳清粉的水分含量、硫代巴比妥酸(TBA)值和羟甲基糠醛(HMF)含量均随贮藏时间的延长而显著增加(P＜0.05),色差随贮藏时间的延长有所波动,但总体呈现上升趋势,而免疫球蛋白(IgG)的含量在60d内逐渐降低,60d后其下降不显著;在25℃、50％ RH条件下PET包装的初乳清粉的货架期为99.40d.     

载板蓝根多糖MPEG-PLA聚合物微球对小鼠免疫力的影响

为研究载板蓝根多糖微球的免疫增强作用。利用实验室制备的载板蓝根多糖MPEG-PLA聚合物微球免疫小鼠,并测定小鼠血液的常规指标,即白细胞和淋巴细胞总数以及总IgG水平和腹腔巨噬细胞吞噬能力等免疫指标。结果表明：在给药后第7天和第14天,载板蓝根多糖微球40、20、10 mg组和板蓝根多糖组小鼠的各项免疫指标均高于空白微球组和生理盐水组,并且载板蓝根多糖微球各组在第14天时的免疫效果与第7天时的免疫效果相比较差异不显著,其中,微球组中以载板蓝根多糖微球20 mg组的免疫效果为最好,在第7天和第14天时小鼠的白细胞和淋巴细胞总数分别为(0.96±0.46)和(11.03±0.35)以及(8.73±0.51)和(9.44±0.42),总IgG水平为(33.541±0.458)和(32.713±0.481),巨噬细胞吞噬指数和吞噬百分率则分别达到(56.00±2.16)和(1.61±0.04)。载板蓝根多糖微球具有一定的免疫增强作用,虽然微球的短期免疫效果均低于纯板蓝根多糖,但是可达到长效免疫。     

牛IgG Fc受体Ⅱ胞外区在大肠杆菌中的表达及特性鉴定

从牛肺巨噬细胞cDNA文库中扩增编码牛IgGFc受体Ⅱ(boFcγRⅡ)胞外区基因,PCR产物经EcoRⅠ和HindⅢ双酶切后,插入原核表达载体pET28a的T7启动子下游,构建重组表达质粒pETboRⅡ。以pETboRⅡ转化大肠杆菌BL21(DE3),IPTG诱导表达。SDS-PAGE分析诱导表达菌体可见分子量约为27kDa的蛋白条带,该表达蛋白在细胞质中主要以不溶性包涵体形式存在。Westernblotting和Dotblot检测表明,boFcγRⅡ重组蛋白在变性条件下可与牛IgG特异结合,提示牛FcγRⅡ胞外区可能存在IgG线性结合表位。     

Efficacy of an oral live vaccine for veterinary use against pseudotuberculosis

Pseudotuberculosis, an infection caused by the ubiquitous enteropathogenic bacterium Yersinia pseudotuberculosis, is a recurrent veterinary problem in livestock and zoo animals. The only vaccine currently available in zoos is Pseudovac (a mixture of killed strains of various serotypes), but its efficacy is not well established. We show here that Pseudovac does not protect guinea pigs against a severe Y. pseudotuberculosis infection. We thus evaluated the possibility of using a live attenuated Y. pseudotuberculosis strain (IP32680) as an oral vaccine against animal pseudotuberculosis. We report that IP32680 is avirulent for guinea pigs and induces a strong IgG response against various serotypes of Y. pseudotuberculosis. One and two oral inoculations of IP32680 provided 50% and 83% protection, respectively against a severe infection with a highly pathogenic strain. The avirulent Y. pseudotuberculosis IP32680 is therefore much more protective than Pseudovac and may represent a valuable oral vaccine against pseudotuberculosis in zoo animals.