微量Bradford法测定提纯禽结核菌素蛋白含量

研究用96孔微量板Bradford法测定提纯禽结核菌素蛋白含量。通过不同浓度的待测禽结核菌素与考马斯亮蓝G-250溶液混合后，在595nm光波下测定各孔样品的OD值。微量Bradford法检测禽结核菌素的线性范围是50—1000μg／mL，相关系数为0．999，最低定量限为50μg／mL。试验用3支禽结核菌素国际参照品，分别配制成125、250、500μg／mL三种不同浓度，每支每种浓度测定4次，计算实际测得的蛋白质浓度。结果显示，批内相对标准偏差为1．6％～4．4％；批间相对标准偏差0．7％-2．2％。研究结果表明微量Bradford法可作为测定禽结核菌素蛋白含量的一种方法。     

Domestication potential of Marula (Sclerocarya birrea subsp caffra) in South Africa and Namibia: 2. Phenotypic variation in nut and kernel traits

As part of a wider study characterizing tree-to-tree variation in fruit traits as a pre-requisite for cultivar development, fruits were collected from each of 63 marula (Sclerocarya birrea) trees in Bushbuckridge, South Africa and from 55 trees from the North Central Region of Namibia. The nuts were removed from the fruit flesh, and the kernels extracted, counted and weighed individually to determine the patterns of dry matter partitioning among the nut components (shell and kernel) of different trees. Mean nut, shell and kernel mass were not significantly different between the two countries. Between sites in South Africa there were highly significant differences in mean nut mass, shell mass, kernel mass and kernel number. In Namibia, there were highly significant differences between geographic areas in mean shell mass, kernel mass and kernel number, but not in nut mass. These differences had considerable impacts on shell:kernel ratios (8.0 - 15.4). In South Africa, mean kernel mass was significantly greater in fruits from farmers fields (0.42 g) than from communal land (0.30 g) or natural woodland (0.32 g). Within all sites, in both South Africa and Namibia, there was highly significant and continuous variation between individual trees in nut mass (South Africa = 2.3 - 7.1 g; Namibia = 2.7 - 6.4 g) and kernel mass (South Africa = 0.09 - 0.55 g; Namibia = 0.01 - 0.92 g). The small and valuable kernels constitute a small part of the nut (Namibia = 6.1- 11.1%; South Africa = 7.6 - 10.7%). There can be 4 kernels per nut, but even within the fruits of the same tree, kernel number can vary between 0-4, suggesting variation in pollination success, in addition to genetic variation. The nuts and kernels of the Namibian trees were compared with the fruits from one superior tree (Namibian Wonder: nuts = 10.9 g; kernels = 1.1 g). Oil content (%) and oil yield (g/fruit) also differed significantly between trees (44.7 - 72.3% and 8.0 - 53.0 g/fruit). The percentage frequency distribution of kernel mass was skewed from trees in farmers fields in South Africa and in some sites in Namibia, suggesting a level of anthropogenic selection. It is concluded that there is great potential for the development of cultivars for kernel traits, but there is also a need to determine how to increase the proportion of nuts with four kernels, perhaps through improved pollination success.     

IS1245 RFLP-based genotyping study of Mycobacterium avium subsp. hominissuis isolates from pigs and humans

Mycobacterium avium subsp. hominissuis, ubiquitous environmental mycobacterium, is an opportunistic pathogen that is regularly isolated from pigs and humans in Slovenia. Genetic diversity of 114 isolates from pigs (n = 57) and humans (n = 57), identified by means of bacteriology, DNA–RNA hybridization techniques, IS901 PCR and IS1245 PCR, was investigated in this study, using IS1245 restriction fragment length polymorphism (RFLP) analysis.Identical IS1245 RFLP profiles were found in isolates from pigs, isolates from humans and isolates from both origins. The proportion of clustered isolates varied as it depended on the similarity level (100% and 75%) chosen for the cluster limits. Using IS1245 RFLP, it was possible to detect monoclonal, polyclonal and recent infections and to monitor the genetic variability of the strains from individual patients.Our findings indicate the environment as the source of infection for both pigs and humans. The questions about the possibility of intra- and inter-species transmission of infection remain to be answered.     

Bovine-associated MRSA ST398 in The Netherlands

During routinely screening (50.000 milk samples on an annual basis) 14 MRSA ST398 strains were identified in the period of January 2008 to September 2008 in 14 different dairy herds located in the provinces Overijssel and Gelderland, The Netherlands. Molecular analysis was performed by Cfr9I PFGE, ST398-specific diagnostic PCR, spa typing, SCCmec typing and Panton-Valentine Leukocidin (PVL) gene PCR. The molecular analyses of 14 MRSA (one MRSA strain per herd) strains revealed that all strains belong to ST398 with 3 closely related spa types (t011, t108 and t889, all commonly found in pigs) and carry 2 different SCCmec types, IVa and V. All MRSA strains were resistant to two or more classes of antibiotics and also PVL negative. The majority of farms (n = 9, 64%) harboured combined livestock with both cows and pigs present. Our study contributes to the growing evidence that MRSA ST398 is transmitted among various animal species and can be considered as an etiological agent of mastitis in dairy cows.     

Microcosm experiments on the development of different plant parasitic nematode fauna in two soils from the Soudanese-Sahelian zone of West Africa

To test the hypothesis that the structure of plant parasitic nematode communities is affected by soil characteristics, experiments were conducted in a greenhouse with two soils with different physical and chemical characteristics and land management histories (fallow and a cultivated field) from adjacent plots. The cultivated soil was more sandy and had lower organic matter and nutrient contents than the fallow soil. Four nematode assemblages of Scutellonema cavenessi, Helicotylenchus dihystera and Tylenchorhynchus gladiolatus were inoculated in the soils. The pot experiment was conducted on millet during 2 months. Multiplication rates of H. dihystera were not significantly different in the two soils. T. gladiolatus had a lower multiplication rate in the fine-textured soil. S. cavenessi seemed to reproduce better in the coarse-textured soil when inoculated in low density with H. dihystera. The presence of plant parasitic nematodes in the cultivated soil caused a significant decrease of millet biomass, whereas plants in the fallow soil were less sensitive to nematode damage and were only affected when the soil was inoculated with T. gladiolatus alone. This experiment did not explain the distribution of plant parasitic species observed in the field. However, parameters other than the presence of a favourable host plant and micro-climatic conditions were found to induce differences in the reproductive rates of several species of plant parasitic nematodes. Received: 14 January 1996     

Design of Nonionic Surfactants for Supercritical Carbon Dioxide

Interfacially active block copolymer amphiphiles have been synthesized and their self-assembly into micelles in supercritical carbon dioxide (CO2) has been demonstrated with small-angle neutron scattering (SANS). These materials establish the design criteria for molecularly engineered surfactants that can stabilize and disperse otherwise insoluble matter into a CO2 continuous phase. Polystyrene-b-poly(1,1-dihydroperfluorooctyl acrylate) copolymers self-assembled into polydisperse core-shell-type micelles as a result of the disparate solubility characteristics of the different block segments in CO2. These nonionic surfactants for CO2 were shown by SANS to be capable of emulsifying up to 20 percent by weight of a CO2-insoluble hydrocarbon into CO2. This result demonstrates the efficacy of surfactant-modified CO2 in reducing the large volumes of organic and halogenated solvent waste streams released into our environment by solvent-intensive manufacturing and process industries.     

Unusual ovarian activity in a mare preceding the development of an ovarian granulosa cell tumour

An 8-year-old mare, with a foal at foot, was inseminated on foal heat with frozen semen, with the resultant pregnancy lost between days 34 and 41. The right ovary developed a large anovulatory follicle that was non-responsive to multiple doses of ovulating agents. The follicle eventually appeared to luteinise, although plasma progesterone concentrations did not reflect this. Another follicle developed, responded to GnRH and resulted in a pregnancy from frozen semen that went to term with a healthy foal. When the mare was examined after foaling, the structure on the right ovary appeared to be a granulosa cell tumour; the left ovary was smaller than normal and non-functional. Surgical removal of the right ovary before increasing photoperiod resulted in a return to function of the left ovary and a pregnancy to frozen semen on the second cycle following removal. Figures showing concentrations of inhibin, progesterone, androstenedione, oestradiol and testosterone are presented for this entire period. Unusual ovarian activity in the mare might be a prelude to the development of a granulosa cell tumour.     

Feasibility of selection for traits associated with cold tolerance in rice under rapid generation advance method

Summary Variability of the hybrid population can be preserved through generations during the process of rapid generation advance (RGA). Effective selection can be done in an early generation for those traits having high heritability value like growth duration and plant height. But selection is not advantageous for traits having low heritability. Thus, selection for panicle exsertion, cold tolerance at seedling stage and panicle length would not be wise during the early generations. However, in the low temperature areas growth duration and plant height are two important factors which determine varietal suitability. As selection of these two characters are possible through RGA, it should be a standard parctice during RGA to eliminate the plants with long growth duration and select plants with optimum plant height.     

Adipogenic differentiation state-specific gene expression as related to bovine carcass adiposity

Genetic regulation of the site of fat deposition is not well defined. The objective of this study was to investigate adipogenic differentiation state-specific gene expression in feedlot cattle (>75% Angus; <25% Simmental parentage) of varying adipose accretion patterns. Four groups of 4 steers were selected via ultrasound for the following adipose tissue characteristics: low subcutaneous-low intramuscular (LSQ-LIM), low subcutaneous-high intramuscular (LSQ-HIM), high subcutaneous-low intramuscular (HSQ-LIM), and high subcutaneous-high intramuscular (HSQ-HIM). Adipose tissue from the subcutaneous (SQ) and intramuscular (IM) depots was collected at slaughter. The relative expression of adipogenic genes was evaluated using quantitative PCR. Data were analyzed using the mixed model of SAS, and gene expression data were analyzed using covariate analysis with ribosomal protein L19 as the covariate. No interactions (P > 0.10) were observed between IM and SQ adipose tissue depots for any of the variables measured. Therefore, only the main effects of high and low accretion within a depot and the effects of depot are reported. Steers with LIM had smaller mean diameter IM adipocytes (P < 0.001) than HIM steers. Steers with HSQ had larger mean diameter SQ adipocytes (P < 0.001) than LSQ. However, there were no differences (P > 0.10) in any of the genes measured due to high or low adipose accretion. Preadipogenic delta-like kinase1 mRNA was greater in the IM than the SQ adipose tissue; conversely, differentiating and adipogenic genes, lipoprotein lipase, PPARγ, fatty acid synthetase, and fatty acid binding protein 4 were greater (P < 0.001) in the SQ than the IM depot. Intramuscular adipocytes were smaller than SQ adipocytes and had greater expression of the preadipogenic gene, indicating that more hyperplasia was occurring. Meanwhile, SQ adipose tissue contained much larger (P < 0.001) adipocytes that had a greater expression (P < 0.001) of differentiating and adipogenic genes than did the IM adipose tissue, indicating more cells were undergoing differentiation and hypertrophy. Adipogenic differentiation state-specific gene expression was not different in cattle with various phenotypes, but adipogenesis in the SQ and IM adipose tissues seems to occur independently.     

MIRU-VNTR typing of Mycobacterium avium in animals and humans: heterogeneity of Mycobacterium avium subsp. hominissuis versus homogeneity of Mycobacterium avium subsp. avium strains

Epidemiological studies on Mycobacterium avium are requisite for revealing infection sources and disease transmission. They are based upon genotyping methods like RFLP and MIRU–VNTR. In our study, MIRU–VNTR typing was applied to 121 previously RFLP typed M. avium field isolates to compare the discriminatory power of both methods. The applicability of MIRU–VNTR typing was studied for isolates from a limited geographic area, namely 41 M. avium subsp. avium and 80 M. avium subsp. hominissuis isolates. Among the former, exhibiting 12 IS901 RFLP types, five MIRU–VNTR types were found with discriminatory index (DI) of 0.716. Among the latter, exhibiting 56 IS1245 RFLP types, 18 MIRU–VNTR types were found with DI of 0.866. Concomitant use of both methods increased DI to 0.981 and 0.995, respectively. MIRU–VNTR typing employing the selected markers provided discernible discrimination among M. avium subsp. hominissuis isolates, but more discriminative markers are needed for M. avium subsp. avium isolates.