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1.
Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR-based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species-specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry.  相似文献   
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OBJECTIVE: To characterize generation of reactive oxygen species (ROS) by equine spermatozoa. SAMPLE POPULATION: Multiple semen samples collected from 9 stallions. PROCEDURE: Equine spermatozoa were separated from seminal plasma on a discontinuous polyvinylpyrrolidone (PVP)-coated silica gradient and resuspended in a modified Tyrode albumin-lactate-pyruvate medium. Amount of hydrogen peroxide (H2O2) generated was assayed by use of a 1-step fluorometric assay, using 10-acetyl-3,7-dihydroxyphenoxazine as a probe for detection of H2O2 in a microplate assay format. Concentration of H2O2 was determined by use of a fluorescence microplate reader. RESULTS: Amount of H2O2 generated increased significantly with time and spermatozoa concentration for live and flash-frozen spermatozoa, and amount of H2O2 generated was significantly greater for flash-frozen than for live spermatozoa. Addition of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) significantly increased generation of H2O2 by live and flash-frozen spermatozoa. Addition of a calcium ionophore also significantly increased the amount of H2O2 generated by live spermatozoa but did not have an effect on amount of H2O2 generated by flash-frozen spermatozoa. Abnormal equine spermatozoa generated significantly greater amounts of H2O2 than did normal spermatozoa. CONCLUSIONS AND CLINICAL RELEVANCE: Equine spermatozoa generate ROS in vitro, possibly via a NADPH-oxidase reaction. Spermatozoa damaged during flash-freezing or morphologically abnormal spermatozoa generated significantly greater amounts of ROS than did live or morphologically normal spermatozoa. Damaged and abnormal spermatozoa generate greater amounts of ROS that may contribute to reduced fertility or problems related to semen preservation.  相似文献   
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We have constructed a physical map of the human genome by using a panel of 90 whole-genome radiation hybrids (the TNG panel) in conjunction with 40,322 sequence-tagged sites (STSs) derived from random genomic sequences as well as expressed sequences. Of 36,678 STSs on the TNG radiation hybrid map, only 3604 (9.8%) were absent from the unassembled draft sequence of the human genome. Of 20,030 STSs ordered on the TNG map as well as the assembled human genome draft sequence and the Celera assembled human genome sequence, 36% of the STSs had a discrepant order between the working draft sequence and the Celera sequence. The TNG map order was identical to one of the two sequence orders in 60% of these discrepant cases.  相似文献   
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ABSTRACT

In

-situ sorbent amendment is a relatively low-cost, low-impact approach for remediation of soil contaminated with heavy metals (HMs), and thus is considered a way to be favored in developing countries. In this study, materials of non-hazardous, alkaline agronomic and industrial by-products were used as sorbents to explore their capacity of in situ immobilization of multiple HMs in mining-impacted arable soil. These sorbents included fly ash (FA), biochar (BC) and apatite (AP) and they were implemented with varying ratios of combinations. Results of soil microcosm tests showed that after incubation for 90 days, concentrations of Pb, Zn, and Cd in their exchangeable forms determined by a sequential extraction method significantly decreased in amended soils, as opposed to the unamended control. Of the five sets of amendments, the composite of FA, BC, and AP resulted in the maximum reduction (up to 80%) in the mobility of Pb, Zn, and Cd in soils. The mechanisms underlying the immobilization of HMs in amended soils might involve processes of surface precipitation, ion exchange and complexation, in which the physicochemical properties of sorbent materials played an important role. The immobilization efficacy of sorbent amendments on HMs in soil was further supported by pot experiments in which significant inhibition of HM accumulation in the belowground and aboveground tissues of maize was observed after 50-day cultivation in amended soils as compared with control soil. Together, these results suggest that the application of cost-saving and environmentally friendly materials derived from wastes as sorbents to remediate soils contaminated with multiple HMs is promising for developing countries like Vietnam.  相似文献   
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The orb web is a spectacular evolutionary innovation that enables spiders to catch flying prey. This elegant, geometric structure is woven with silk fibers that are renowned for their superior mechanical properties. We used silk gland expression libraries to address a long-standing controversy concerning the evolution of the orb-web architecture. Contrary to the view that the orb-web design evolved multiple times, we found that the distribution and phylogeny of silk proteins support a single, ancient origin of the orb web at least 136 million years ago. Furthermore, we substantially expanded the repository of silk sequences that can be used for the synthesis of high-performance biomaterials.  相似文献   
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Sugarcane is an important cash crop in Vietnam and has been widely promoted at national and provincial level. In 2006, a new disease was discovered in sugarcane in the Nghean Tate&Lyle Sugar Mill in Nghean Province in north-central Vietnam. The key symptoms were the formation of green grassy shoots around the base of mature stools. We applied nested polymerase chain reaction (PCR) using P1/P7 and R16F2n/R16R2 for detection and characterization of phytoplasma from the symptomatic tissues. PCR products of the expected size (approx. 1200?bp) were obtained from the 16S rDNA of the phytoplasma. The restriction fragment length polymorphism profiles indicated that all samples were infected by the same phytoplasma. Phylogenetic analysis confirmed that the SCGS phytoplasma from Vietnam belong to the 16SrXI group, formerly Rice Yellow Dwarf group.  相似文献   
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Dairy production is not traditional in Vietnam. The farmers have little practical knowledge and udder health control is generally lacking. In order to give the farmers appropriate advice, knowledge about the distribution of udder pathogens is crucial. The aim of the study was to investigate the prevalence of sub-clinical mastitis and to identify udder pathogens isolated from smallholder dairy herds in Southern Vietnam. Twenty farms with a herd somatic cell count (SCC) ranging from low (≤400?×?103?cells/mL) to high (>400?×?103?cells/mL) were randomly selected. Milk samples were collected from 458 quarters of 115 clinically healthy cows. SCC was analyzed on farm by a portable cell counter. Bacteriological samples were taken using Mastistrip© cassettes and sent to Sweden for examination. For all herds the mean herd SCC was 632?×?103/mL milk. The prevalence of subclinical mastitis at quarter SCC basis was 63.2 % and at cow basis 88.6 %. Only 40 % of all cows were bacteriologically negative in all quarters. Streptococcus agalactiae was the most commonly found bacteria species, isolated from 96 of the 458 quarter samples, in 13 of the 20 farms. The results indicate pronounced subclinical mastitis problems among the dairy cows in this region mainly due to infections with S. agalactiae. The high prevalence of this highly contagious pathogen is probably attributable to the generally poor milking hygiene and low awareness of proper measures to prevent occurrence and spread of udder infections. A strict, targeted action program for the herds in this area is required in order to lower the prevalence of subclinical mastitis.  相似文献   
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