以超滤技术浓缩猪细小病毒含毒细胞液的试验

在研制猪细小病毒灭活疫苗时,应用了超滤浓缩技术,提高了合毒细胞培养液中的抗原含量。为了保证浓缩的流量,减少对滤膜的污染,对含毒细胞培养液作了预处理一离心及微滤,以便除去细胞培养液中的细胞碎片、蛋白质等大分子物质及固形微粒。超滤时选用50000Da滤膜,在20psig压力下,经过5倍浓缩的PPV细胞培养液,血凝滴度提高2个滴度以上。病毒含量测定表明,TCID50／0．2mL由107左右升至109以上,以其配制疫苗,能显著地提高疫苗的免疫原性。超滤技术具有易于操作、高效、分离精度高、没有二次污染等优点,可以根据需要选择不同的浓缩浓度,对保证疫苗的质量具有重要作用。     

2种三叶草杂种F1代胚性愈伤组织筛选

以高加索三叶草(Trifolium ambiguum Bieb.)与白三叶(Trifolium.repens L.)杂交后胚离体培养产生的F₁无菌苗的茎段为外植体,筛选诱导愈伤组织的最适培养基,将F₁茎段产生的愈伤组织经过继代后转移到分化培养基中,研究不同浓度激素组合对体细胞胚胎产生的影响。结果表明:MS+6-BA+NAA是诱导F₁茎段愈伤组织的适宜培养基;MS+2,4-D+6-BA+NAA+KT+CH为F₁茎段愈伤组织的适宜分化培养基;对分化后的胚性愈伤进行组织学观察,可见其表面形成许多瘤状突起,即胚性细胞团,它们可以继续发育形成体细胞胚,为快速得到数量较多的杂种F₁再生苗奠定基础。     

Demographics of cattle positive for Mycobacterium avium subspecies paratuberculosis by faecal culture,from submissions to the Cork Regional Veterinary Laboratory

The demography of bovine infections caused by Mycobacterium avium subspecies paratuberculosis (MAP) in Ireland is poorly defined. The objective of this study was to describe the demographics of cattle positive to MAP on faecal culture, based on submissions to the Cork Regional Veterinary Laboratory (Cork RVL) from 1994 to 2006. The study focused on all available faecal samples from adult cattle with non-responsive chronic diarrhoea that were submitted by private veterinary practitioners to Cork RVL for MAP culture. For each MAP-positive by faecal culture animal, data were collated from Cork RVL and Cattle Movement Monitoring Scheme (CMMS) records. Johne's disease (JD) was confirmed in 110 animals from 86 herds by the Cork RVL between 1994 and 2006, with a rate of positive cases between 15% and 18% over last four years of the study. Two breeds (Holstein/Friesian or Limousin) made up 78% of submissions. Movements were assessed for the 57 study animals with available movement information, 90% died within one year of the test and 26% tested positive in the herd they were born into. The study provides preliminary information about movement trends and demographics of animals with MAP positive submissions. Although the study area is restricted, it includes the most intensive (and economically-important) dairy region in Ireland. The demographics of JD infection from the study area are in agreement with international reports. Further work is required to determine demographic trends, incidence and prevalence of JD throughout Ireland. It is hoped this work may contribute to the development of a surveillance strategy for MAP by regional veterinary laboratories.     

蒙古族草原文化传统的生态学内涵

蒙古民族由于长期的游牧生活,形成了独特的文化传统。从蒙古草原文化传统对于草原生态功能维护和草地畜牧业发展的贡献入手,分析了蒙古草原文化传统的特点,认为蒙古草原文化中爱护家畜、维护环境的朴素感情是人与自然和谐相处的精神体现,为当今实施可持续发展模式提供了良好的思想基础,对于遵循科学发展观,寻求草原地区的发展模式和畜牧业的产业化途径,具有重要的启示。     

复杂盐碱条件对星星草种子萌发的影响

将2种中性盐NaCl、Na2SO4及2种碱性盐Na2CO3、NaHCO3按不同比例混合。模拟出24种盐度和碱度各不相同的复杂盐碱条件,并用以处理星星草种子,测定其发芽率、发芽势。结果表明,不同碱度的盐分组成对星星草种子萌发的影响基本一致,即均随着盐浓度的增高发芽延迟,发芽率降低,而且均在盐浓度超过300 mmol/L时发芽率降至零。对各胁迫因素与发芽率之间进行统计学分析,结果表明,盐浓度是影响星星草种子萌发的决定性因素,其他因素的作用很小。本实验结果表明,吸水是决定星星草种子萌动与否的关键,由于盐浓度决定水势进而决定种子吸水,所以盐浓度表现为影响种子萌发的关键因素。由于pH值、缓冲量等代表碱度的指标与水势没有直接关系,所以对星星草种子萌动阶段没有决定性影响。     

脂肪细胞培养研究现状

脂肪细胞培养为研究脂肪代谢紊乱疾病提供了一种有效的途径。基质血管成分系统的研究形成了完整的前脂肪细胞理论,随后脂肪细胞原代培养技术得到了广泛的研究,国外各种家畜的脂肪细胞培养模型得以建立,而国内在脂肪细胞培养方面的研究起步较晚,且多集中于人及鼠的脂肪细胞培养。脂肪细胞的传代培养在单一因子对脂肪细胞的功能研究上有一定的优越性,尤其对不宜于在活体采样的人及动物。脂肪细胞培养的另一个研究方向是脂肪细胞与其他组织细胞的联合培养,对研究机体内各种组织细胞间的相互作用提供了良好的基础。     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.     

过瘤胃保护蛋氨酸稳定性及其评定方法的研究

本研究应用双外流连续培养系统和瘤胃尼龙袋法检验动物油包被蛋氨酸在瘤胃中的稳定性。根据动物油添加比例不同(分别添加20%、30%和40%),保护性蛋氨酸分为过瘤胃保护蛋氨酸I(RPMet-I)、过瘤胃保护蛋氨酸II(RPMet-II)和过瘤胃保护蛋氨酸III(RPMet-III)。试验结果表明:与未包被蛋氨酸相比,动物油包被蛋氨酸在瘤胃中的释放率显著降低(P<0.05);动物油添加比例不同对于过瘤胃保护蛋氨酸的稳定性影响显著(P<0.05)。不同形式保护性蛋氨酸在双外流连续培养系统中的降解趋势与在瘤胃中的降解趋势相似,且应用双外流连续培养系统测定各个时间点保护性蛋氨酸的释放率简单、稳定,与瘤胃尼龙袋法相比各个时间点的释放率均存在强相关回归关系(n=5,P<0.01),因此该系统适宜于评定保护性氨基酸的稳定性。     

Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.