Real time polymerase chain reaction for rapid and quantitative determination of Cystofilobasidium infirmominiatum on the surfaces of apple,pear, and sweet cherry fruit

The objectives of this study were to develop primers and a real time PCR protocol for the postharvest biocontrol yeast Cystofilobasidium infirmominiatum (Cim). The application of this technology was developed to quantify Cim on the surfaces of apple, two pear cultivars, and sweet cherry fruit treated over a range of concentrations. Statistically significant relationships were observed between Cim DNA on fruit surfaces, expressed as μg/m², and CFU/L of dip suspensions for apple, pear, and sweet cherry. In addition, the relationship for each fruit was significantly different from the other three fruits. Threshold values of concentrations of Cim DNA on the fruit surface were calculated based on regression equations and a dose of 2.0 × 10¹¹ CFU/L of dip suspension, the dose for optimum decay control, and were 4.8, 7.0, 16.5, and 25.2 μg/m² for Bosc pear, Lapins sweet cherry, d’Anjou pear, and Golden Delicious apple, respectively. Monitoring Cim DNA concentration on fruit surfaces will assure that Cim is being properly applied to fruit and that a sufficient number of cells are present for optimum decay control.     

中棉所63棉铃对位叶光合生理性能的杂种优势研究

以杂交棉中棉所63及其亲本为材料,在棉铃发育时期对棉铃对位叶的生理和光合性能指标进行测定,探索中棉所63获得高产的杂种优势形成机理。结果表明:1)中棉所63产量优势显著。单株结铃数比两亲本分别高21.52%,43.28%,铃重高2.42%,19.31%,叶面积载铃量高22.39%,22.30%。2)中棉所63在棉铃发育后期棉铃对位叶生理指标表现出显著的杂种优势。棉铃对位叶叶面积分别比亲本高26.60%,59.41%,并且中棉所63棉铃对位叶保持较高水平的SOD活性,使其生理代谢明显提高;与此同时,中棉所63的MDA含量显著低于亲本,细胞膜受损较轻,延缓了对位叶的衰老。3)中棉所63在棉铃发育后期棉铃对位叶光合指标表现出显著的杂种优势。在棉铃发育后期,中棉所63叶绿素含量、SPS活性的高水平,能够维持高效率的光合作用,积累更多的光合产物,从而使中棉所63获得较高的单株结铃数和铃重,最终获得高产。     

Effects of cytokines from thymocytes and thymic stromal cells on chicken intrathymic T cell development

We have studied the ability of thymic stromal cells (TSC) and thymocytes to produce cytokines and the involvement of cytokines in intrathymic T cell development. When thymocytes were co-cultured with thymic stromal cells in absence of direct contact and mitogenic stimulation, induction of thymocyte proliferation was observed. Supernatants of cultured stromal cells (TSC-CS) promoted a high proliferative response on CD3- thymocytes but had little effect on CD3+ thymocytes. These results indicate that stromal cells have produced a cytokine which can induce immature thymocyte proliferation. Moreover, stromal cells express the MRNA for stem cell factor (SCF) and c-kit (the receptor for SCF) was detected on CD3- thymocytes but not on CD3+ thymocytes. Since SCF can enhance the proliferation of immature thymocytes in synergy with IL-7 in mammals, there is a possibility that chicken stromal cells may produce a IL-7-like factor. Thymocytes have clearly expressed interferon (IFN)-gamma. In contrast, thymic stromal cells showed no detectable expression of IFN-gamma. CD3+ thymocytes express IFN-gamma MRNA more strongly than CD3 thymocytes, suggesting that IFN-gamma from thymocytes may operate on stromal cells and then may indirectly induce clonal elimination of CD3+ cells on stromal cells. The expression of these cytokines and receptors by thymic stromal cells and thymocyte subpopulations suggests that these cytokines participate in paracrine interactions between these cell populations during thymocyte differentiation.     

Analysis of the role of forest,biomass policy legislation and other factors that may affect the future of Kenya’s forests: Use of Japanese forestry as a model

Forests are one of the most important ecosystems on earth that require careful management, conservation, and sustainable exploitation. As countries have their own guideline systems, each may learn and borrow from one another’s experience. One of such countries is Japan, which has elaborate forest policies, and rich in forest cover (67%) with its forest history dating many years back. On the other hand, Kenya, with a forest cover of just 7% and its policies demonstrating notable weaknesses, has a lot to learn from Japan. Therefore, we have attempted to do a comparative analysis of forest policies, technologies, and management practices between Kenya and Japan. Results indicate that Kenya’s forest policies do not place adequate emphasis on silvicultural practices and the establishment of forest plantations, and rarely focus on sustainable biomass utilization—factors that contributed significantly to forest growth and development in Japan. Additionally, policy legislation, revision, and implementation have not been given the deserved priority in Kenya. We conclude that Kenyan forest policy would benefit from both revision and thorough implementation. We also discuss the role of indirect factors such as economic growth and availability of nonwood-based energy sources in the future of Kenya’s forests.     

新疆伊犁某牛场不同生长期牛粪源大肠杆菌耐药性调查

为了调查新疆伊犁某牛场不同生长期牛粪源大肠杆菌对临床常用抗菌药物的耐药情况。从该牛场分别采集育肥牛、繁育牛、加育牛和未繁育牛的粪样。采用微量肉汤稀释法,对分离出的大肠杆菌进行最小抑菌浓度测定。结果表明,育肥牛、繁育牛、加育牛和未繁育牛粪源大肠杆菌分离率均为100%,不同生长期的牛源大肠杆菌对氨苄西林均耐药,耐药率分别为4.0%、8.6%、6.7%、20.0%;对安普霉素都较为敏感。繁育牛和加育牛粪源大肠杆菌耐药种类较多,未繁育牛粪源大肠杆菌对比其他生长期牛粪源菌耐药较严重,不同生长期的牛粪源大肠杆菌多重耐药情况不太严重,主要为0耐、1耐;新疆伊犁该牛场对临床常用抗菌药物的耐药程度不同,耐药程度虽然不高,但在养殖生产中需注意合理用药。     

Improvement of DNA extraction method for dried blood spots and comparison of four PCR methods for detection of Babesia gibsoni (Asian genotype) infection in canine blood samples

To eradicate canine babesiosis in epidemic areas, mass-screening of the infection situation of Babesia gibsoni including occult infection is necessary. The development of cost-effective method for storage and transport of blood samples is required. A highly efficient DNA extraction procedure from dried blood spots (DBS) onto Whatman 3MM filter paper was developed for the diagnosis of B. gibsoni infection in dog by PCR. In 3 extraction methods, Chelex-based method in combination with saponin washing and phenol-chloroform-isoamyl alcohol extraction (Saponin-PCI method) provided the best results. Sensitivity of the 4 previously described PCR methods for detection of B. gibsoni infection was also compared using serially diluted blood samples of B. gibsoni-infected dogs. The PCR method using Gib599F/Gib1270R primer pair provided the best performance. To evaluate the stability of DNA in DBS, DBS of B. gibsoni-infected dogs stored at room temperature for 2 months. The stability was superior to whole blood samples stored at -20 degrees C for 2 months. This highly efficient DNA extraction method on DBS using Whatman 3MM filter paper has potential to be cost-effective and high performance tool for storage, and molecular diagnosis of clinical blood sample from dog. This procedure in combination with the PCR method using Gib599F/Gib1270R primer pair may greatly assist in diagnosis of B. gibsoni infection in dog populations that are geographically distant.     

Assessing foraging strategies of herbivores in Mediterranean oak woodlands: a review of key issues and selected methodologies

Montados are agro-silvo-pastoral ecosystems, typical of the Southwest Iberian Peninsula, of high socio-economic and conservation importance, where grazing is a dominant activity. Montados are characterized by an open tree canopy of Quercus sp. and a diverse undercover of shrubs and grasslands that constitute the plant food resources for grazing animals. Plant food resources of Montados are highly variable, both spatially and seasonally, in quantity and quality. Reliable and easy to use methods to monitor grazing are necessary to allow proper understanding of foraging strategies of grazing animals and to set sustainable grazing management. We describe the main characteristics of the plant food resources available for grazing animals, striking its variability, and revise the potential of using N-alkanes and saliva proteome methods to assess foraging strategies in Montados. In a scenario of dynamic multiple choices, we discuss the use of n-alkane methodology for the simultaneous estimation of diet composition and voluntary intake and saliva proteome as a mean of increasing the knowledge on diet adjustments.     

Establishment and growth of coniferous seedlings in an altered forest floor after long-term exclusion of deer

We evaluated differences between the forest floors and the establishment and growth of coniferous seedlings in fenced (13 years) and unfenced plots on Mt Ohdaigahara where the sika deer (Cervus nippon) population density is high. Large coniferous seedlings (height > 0.05 m) were less abundant in the unfenced plot, as a result of deer browsing. Small coniferous seedlings (height < 0.05 m), however, were more abundant in the unfenced plot, where most seedlings of Abies homolepis were found on bare ground and those of Picea jezoensis var. hondoensis were found on buttresses and fallen logs. The large area of bare ground in the unfenced plot was caused by deer browsing. Deer therefore have an indirect effect on the emergence and growth of small coniferous seedlings by modifying the forest floor.     

Precision of genetic relationship estimates based on molecular markers

Genetic progress through selection is directly related to the amount of variability present in the population and the quality of genes contributed by the parents. Genetic relationships between lines were studied using DNA marker-based estimates of genetic similarity. A statistical methodology using the width of a confidence interval was developed to determine the number of probes to be surveyed and the precision in the estimation of genetic distance between pairs of cultivars. Precision was affected by type of genetic distance used, the number of cultivars, and amount of genetic diversity present in the studied group. The width of a (1-α)% confidence interval decreased as the number of RFLP fragments increased. Oat and wheat diversity studies were used to illustrate the methodology. This revised version was published online in July 2006 with corrections to the Cover Date.