Divergence of a strain of Pseudomonas aeruginosa during an outbreak of ovine mastitis

Bacterial infections causing mastitis in sheep can result in severe economic losses for farmers. A large survey of milk samples from ewes with mastitis in Sardinia, Italy, indicated an increasing prevalence of Pseudomonas aeruginosa infections. It has been shown previously that during chronic, biofilm-associated infections P. aeruginosa populations diversify. We report the phenotypic and genomic characterisation of two clonal P. aeruginosa isolates (PSE305 and PSE306) from a mastitis infection outbreak, representing distinct colony morphology variants. In addition to pigment production, PSE305 and PSE306 differed in phenotypic characteristics including biofilm formation, utilisation of various carbon and nitrogen sources, twitching motility. We found higher levels of expression of genes associated with biofilm formation (pelB) and twitching motility (flgD) in PSE305, compared to the biofilm and twitching-defective PSE306. Comparative genomics analysis revealed single nucleotide polymorphisms (SNPs) and minor insertion/deletion variations between PSE305 and PSE306, including a SNP mutation in the pilP gene of PSE306. By introducing a wild-type pilP gene we were able to partially complement the defective twitching motility of PSE306. There were also three larger regions of difference between the two genomes, indicating genomic instability. Hence, we have demonstrated that P. aeruginosa population divergence can occur during an outbreak of mastitis, leading to significant variations in phenotype and genotype, and resembling the behaviour of P. aeruginosa during chronic biofilm-associated infections.     

喷雾干燥法浓缩乳清蛋白的工艺参数研究

干酪副产物乳清进行超滤浓缩,将浓缩液喷雾干燥制备成浓缩乳清蛋白,通过对喷雾干燥的进口温度、料液温度、入料流量3个因素进行单因素和正交试验分析,确定了对浓缩液喷雾干燥的最佳工艺参数为：进口温度为140℃,料液温度为30℃,入料流量7%,制得的浓缩乳清蛋白与市售产品进行加热,利用激光粒径分析仪测定产品的粒径分布对比热稳定性,试验制得的浓缩乳清蛋白溶解性、分散性俱佳。     

菠萝皮发醇生产饲料蛋白的工艺条件研究

菠萝皮是菠萝加工产生的副产物,为了将菠萝加工企业废弃的菠萝皮有效利用,实现资源的再生,缓解目前蛋白饲料的不足,进行了本研究.结果表明,采用绿色木霉和产朊假丝酵母混合菌种发酵,添加麸皮20%、硫酸铵3%、尿素2%,料水比11,自然pH值,30℃发酵5 d,发酵后的饲料粗蛋白含量提高到25.38%,经水洗后的粗蛋白含量提高到17.03%,检测的16种氨基酸含量均有很大幅度的提高,其总和从2.89%提高到15.6%,提高率达到439.8%.产品气味芳香,适口性大为改善,可用做牛、羊和鱼等养殖的低蛋白能量饲料.     

马乳脂肪球膜蛋白组鉴定及潜在功能分析

试验旨在研究马乳脂肪球膜（milk fat globule membrane,MFGM）蛋白组成及潜在的生物学功能。选取12匹5岁左右、平均体重为450~500 kg的健康纯血母马作为试验动物,主要饲喂干牧草,所有马匹饲养和管理条件均相同。将12份样品以每组4份进行混合,分离提取产后第60天马乳中MFGM蛋白,进行高分辨质谱仪鉴定和生物信息学分析。结果显示,马MFGM组分中共鉴定出310种表达蛋白,这些蛋白主要参与的生物过程为生物调节、刺激应答、定位、多细胞生物过程、运输、信号、细胞通讯、发育过程、细胞分化和免疫系统过程等;细胞成分主要为胞外区域、膜、囊泡、核仁和线粒体等;分子功能主要为催化活性、蛋白质结合、碳水化合物衍生物结合和小分子结合等。KEGG通路分析表明,MFGM蛋白主要参与血小板活化通路、内吞、脂肪酸生物合成和Ras信号通路等。马乳MFGM蛋白的蛋白质互作网络分析图中共包含215种蛋白质。本研究结果揭示了马乳MFGM蛋白质组的复杂性,为解析其营养和生物学功能提供了科学数据。     

凋亡素基因真核表达载体的构建及在人肿瘤细胞中的表达

为探讨凋亡素对肿瘤的基因治疗效果,构建了凋亡素基因的真核表达载体。将凋亡素基因重组入 ｐｃ Ｄ Ｎ Ａ３ 载体,并通过脂质体介导凋亡素在人喉癌、人肺癌细胞系中表达;通过 Ｒ Ｔ Ｐ Ｃ Ｒ 在转染细胞中检测到了凋亡素的 ｍ Ｒ Ｎ Ａ,这为应用凋亡素进行肿瘤的基因治疗奠定了基础。     

Serine-to-glycine ratios in low-protein diets regulate intramuscular fat by affecting lipid metabolism and myofiber type transition in the skeletal muscle of growing-finishing pigs

Serine and glycine are 2 of the first-affected nonessential amino acids in low crude protein (CP) diets for pigs. Therefore, we explored the effects of different dietary serine-to-glycine ratios on growth performance and lipid metabolism in growing-finishing pigs. A total of 160 crossbred healthy barrows, with a similar body weight of around 59.50 kg, were randomly allotted into 1 of 5 treatments (8 pens per treatment and 4 pigs per pen). The serine-to-glycine ratios of the 5 dietary treatments were as follows: diet A (NORMAL group), 1.18:1 (16% CP); diet B (LOW group), 1.2:1 (12% CP); diet C (S2G1 group), 2:1 (12% CP); diet D (S1G2 group), 1:2 (12% CP); and diet E (S1G1 group), 1:1 (12% CP).We found that the pigs fed a low CP diet (12% CP), when maintaining serine-to-glycine ratio at 1:2 and a total amount of 1.44%, had the same average daily gain as the pigs fed a normal CP diet (16% CP) (P > 0.05), but they had increased intramuscular fat (P < 0.05). Furthermore, they exhibited higher expression of genes involved in lipid oxidation (P < 0.05), which was regulated by modulating methylation levels in the promoters of acyl-CoA oxidase 1 (ACOX1) and acyl-CoA dehydrogenase medium chain (ACADM). When compared with the pigs fed a normal CP diet, these pigs had more oxidative myofibers (P < 0.05), which were regulated by AMPK-PGC-1α and Calcineurin-MEF2/NFAT pathways in a coordinated manner. Our findings suggested that a dietary serine-to-glycine ratio of 1:2 is beneficial for improving meat quality in pigs fed a low CP diet.     

四神叶形吸虫的DNA含量及蛋白质电泳分析

四种叶形吸虫成虫初级精母细胞核ＤＮＡ含量测定表明，寄生于较高等鱼类宿主的中华叶形吸虫和鳗鲡叶形吸虫的ＤＮＡ含量较高（Ｃ值分别为０．２５ｐｇ和０．２６ｐｇ），而寄生于较低等鱼类宿主的鲶叶形吸虫和巴氏叶形吸虫的ＤＮＡ含量较低（Ｃ值分别为０．２１ｐｇ和０．２０ｐｇ）。这四种叶形吸虫成虫全蛋白质浓度梯度凝胶电泳分析进一步显示，中华叶形吸虫与鳗鲡叶形吸虫构成一近缘亚群（Ｓｍ＝０．６５７９，Ｄ＝０．４１８７），而鲶叶形吸虫和巴氏叶形吸虫构成另一近缘亚群（Ｓｍ＝０．６１７６，Ｄ＝０．４８１９）；这两个亚群间的相似系数值（Ｓｍ）小于０．５３，遗传距离值（Ｄ）大于０．６４。以上结果从分子水平上再次证实将叶形吸虫属划分为两个亚属是有根据的。     

猪戊型肝炎病毒ORF2片段克隆及原核表达

设计套式引物，内套引物含有BamHⅠ／XhoⅡ酶切位点，采用RT-PCR技术扩增猪戊型肝炎病毒（UEV）结构蛋白基因ORF2部分片段，长度为634bp，将其克隆于PGEX-T载体，测序结果表明插入的片段属于戊型肝炎病毒ORF2部分，将该片段插入PproEXHTb表达载体，经酶切鉴定证实获得了重组表达质粒。将重组质粒转化大肠杆菌BL21，经1mmol／L IPTG诱导，得到表达，表达蛋白分子量为27Ku，以包涵体形式存在。Western blot分析表明，该蛋白可以与猪戊型肝炎阳性血清反应，表明该蛋白具有良好的反应原性。     

热应激蛋白对畜禽抗应激机理的研究进展

热应激蛋白(heat shock proteins, HSPs)是机体在应激条件下所产生的一组特异性蛋白质,能使机体迅速适应环境变化。作者简要介绍了HSPs的来源及分类,阐述了HSP70 的基因结构和调控机制,综述了HSPs抗应激功能及在畜禽应激反应中的表达,以期为从分子水平探索HSPs 抗应激机理提供参考。     

Immunopathology of <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> Infection

Heartworm disease caused by Dirofilaria immitis affects canine and feline hosts, with infections occasionally being reported in humans. Studies have shown that both dirofilarial antigens and those derived from its bacterial endosymbiont Wolbachia, interact with the host organism during canine, feline and human infections and participate in the development of the pathology and in the regulation of the host’s immune response. Both innate and acquired immune responses are observed and the development of the acquired response may depend on the host and, or on its parasitological status. This review aims at illustrating current research on the role of both D. immitis and Wolbachia, in the immunology and immunopathology of dirofilariosis.