Field comparison of the interferon-gamma assay and the intradermal tuberculin test for the diagnosis of bovine tuberculosis

An extensive field comparison of the gamma interferon (IFN-gamma) assay and the single intradermal tuberculin test for the diagnosis of bovine tuberculosis was conducted in Australia. The specificity of the IFN-gamma assay was determined by testing more than 6000 cattle from tuberculosis-free herds and varied from 96.2% to 98.1%, depending on the cut-off point chosen to define a positive reactor. For the sensitivity trial, cattle from herds being de-populated because of bovine tuberculosis were examined with both assays. The sensitivity of the IFN-gamma assay was shown to be significantly higher than the single intradermal tuberculin test and varied from 76.8% to 93.6% depending on the method of interpretation. A maximum overall sensitivity of 95.2% was obtained by testing with the IFN-gamma and the tuberculin test in parallel. The superior sensitivity of the IFN-gamma assay and the ability to adjust the sensitivity of the system depending on the task involved, will provide the Australian Tuberculosis Eradication Campaign with a valuable additional test to enable it to accomplish its goals.     

木麻黄不同亲缘间嫁接对嫁接亲和力和花诱导的影响

通过嫁接对木麻黄的亲本进行矮化是开展木麻黄杂交育种的重要技术基础。为了研究木麻黄 不同亲缘间嫁接对嫁接亲和力和花诱导的影响,以木麻黄属的短枝木麻黄（Casuarina equisetifolia）为砧 木,分别以同种的短枝木麻黄,同属不同种的细枝木麻黄（C. cunninghamiana）、粗枝木麻黄（C. glauca） 和不同属的滨海木麻黄（Allocasuarina littoralis）为接穗,开展木麻黄不同亲缘关系间的嫁接研究,研究 种内、种间和属间嫁接对木麻黄嫁接亲和力和花诱导的影响。研究结果表明,木麻黄种内嫁接的亲和力 最高,无性系接穗嫁接成活率高达91.4%,实生苗接穗为87.0%;而种间嫁接的亲和力显著低于种内嫁 接,细枝木麻黄接穗和粗枝木麻黄接穗嫁接成活率分别为36.0% 和45.0%;属间的嫁接未发现有成活的 例子,其嫁接亲和力为0。在嫁接对接穗的花诱导方面,种内嫁接的无性系接穗次年开花率为74.2%,显 著高于种内嫁接的实生苗接穗（42.3%）和种间嫁接的细枝木麻黄（39.8%）和粗枝木麻黄（35.6%）。     

Molecular and Cellular Characterization of Buffalo Bone Marrow‐Derived Mesenchymal Stem Cells

Immune privileged mesenchymal stem cells (MSCs) can differentiate into multiple cell types and possess great potential for human and veterinary regenerative therapies. This study was designed with an objective to isolate, expand and characterize buffalo bone marrow‐derived MSCs (BM‐MSCs) at molecular and cellular level. Buffalo BM‐MSCs were isolated by Ficoll density gradient method and cultured in Dulbecco’s modified Eagle’s medium supplemented with fetal bovine serum (FBS). These cells were characterized through alkaline phosphatase (AP) staining, colony‐forming unit (CFU) assay, mRNA expression analysis (CD 73, CD 90, CD 105, Oct4 and Nanog), immunolocalization along with flow cytometry (Stro 1, CD 73, CD 105, Oct4, Sox2 and Nanog) and in situ hybridization (Oct4 and Sox2). Multilineage differentiation (osteogenic, adipogenic and chondrogenic) was induced in vitro, which was further assessed by specific staining. Buffalo BM‐MSCs have the capacity to form plastic adherent clusters of fibroblast‐like cells and were successfully maintained up to 16^th passage. These cells were AP positive, and further CFU assay confirmed their clonogenic property. RT‐PCR analysis and protein localization study showed that buffalo BM‐MSCs are positive for various cell surface markers and pluripotency markers. Cytoplasmic distribution of mRNA for pluripotency markers in buffalo BM‐MSCs and multilineage differentiation were induced in vitro, which was further assessed by specific staining. To the best of our knowledge, this is the first report of buffalo BM‐MSCs, which suggests that MSCs can be derived and expanded from buffalo bone marrow and can be used after characterization as a novel agent for regenerative therapy.     

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA‐1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA‐1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo‐derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA‐1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA‐1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8–16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA‐1A revealed that it shares 91–98% identity with other mammalian sequences. It can be concluded that higher level of HSPA‐1A mRNA in IVP embryos in comparison with in vivo‐derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA‐1A gene could be used as a stress biomarker during pre‐implantation development.     

Immunopotentiation of a Developed Salmonella enterica serotype Enteritidis Vaccine by Thymulin and Zinc in Meat Chicken Breeders

The humoral immunity, spleen and thymus weight indices, lymphocyte count in the thymus cortex, and granuloma diameter at vaccination sites were assessed in four differently immunopotentiated groups of meat chicken breeders. Breeders in the first two groups were given a killed Salmonella enterica serotype Enteritidis (SE) vaccine subcutaneously at 15 and 19 weeks of age. Breeders in the third and fourth groups were left unvaccinated. Breeders in the first group were further immunopotentiated with zinc and thymulin. Each bird in the first group was given the immunopotentiators intraperitoneally in a volume of 0.1 ml at intervals of 3 days for a period of 3 weeks, starting at 15 weeks of age. At each time, each bird in the first group received thymulin (10 ng) and ZnCl₂ (1 mol/L), using a carboxymethyl cellulose carrier, totalling 90 ng thymulin and 9 mol of ZnCl₂ per bird. Each bird in the first three groups was challenged orally with 6.7×10⁶ cfu/ml of highly virulent SE organisms, at an age of 22 weeks. The first group, which had received zinc and thymulin, had the earliest and highest humoral immune response to SE (p<0.05). This was observed at 2 and 4 weeks after the first vaccination. In addition, the first group had the highest mean thymus weight index, and the highest mean lymphocyte count in the thymus cortex. No significant difference was observed between the first two vaccinated groups in the mean granuloma diameter developed at the two vaccination sites 48 h after administration of the vaccine (p>0.05).     

不同耕作方式对宁南旱地土壤团聚体的影响

在宁南旱区对试验耕地定位实行连年传统翻耕(CT)、隔年免耕—深松(NT→ST)及深松—免耕(ST→NT)三种不同处理。用干筛法和湿筛法对连续3年处理后土壤团聚体的变化进行测定,分析结果表明,干筛法中NT→ST处理在0~40 cm土层中0.25 mm的机械稳定性团聚体比处理前和CT处理分别高33.11%、29.51%;ST→NT处理在0~40 cm土层中0.25 mm机械稳定性团聚体含量比处理前和CT处理分别高40.51%、36.91%。湿筛法中NT→ST处理在0~30 cm土层中0.25 mm的水稳性团聚体含量比处理前高7.33%,NT→ST处理在0~10 cm土层0.25 mm的水稳性团聚体比CT处理高1.47%,而在10~40 cm土层NT→ST处理和CT0.25 mm的水稳性团聚体含量差异不显著;ST→NT处理在0~40 cm土层中0.25 mm水稳性团聚体含量比处理前和CT处理分别高10.75%、10.12%。分析结果同时说明,采用深松—免耕(ST→NT)轮耕方式更有利于土壤团聚体含量和稳定性的增加。