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51.
To achieve stable expression of the heterologous and reporter genes in the Bradyrhizobium genome, we constructed suicide plasmids capable of site-directed genomic integration of the gusA, gfp and nifA genes by homologous recombination into non-essential repeated sequences (RS-α), isolated from B. japonicum strain CPAC7 (SEMIA5080). In this report, we describe the strategies to construct the vectors and their use to obtain mutants with site-specific insertions.  相似文献   
52.
A new chemotype of the aromatic Verbenaceae species Lippia alba Mill. N. E. Br. from southeastern Brazil has recently been shown to have a high content of linalool in the leaf essential oil. Vegetative propagation of this chemotype was conducted at six different locations in Brazil, and the variation of the content and the optical purity of linalool in the oils were verified. Yields (0.6-0.9%, hydrodistillation), chemical composition, linalool content, and optical purity of the oils from all the plants were compared, using GC-FID, GC-MS, chiral chromatography, and retention index calculation. No plant exceeded the matrix in linalool content (46.5 to 90.7%), and the chemical profile of the oils was the same for all the samples. Purification of linalool to a content close to 100% was effected by vacuum distillation of the crude oil. Chiral analysis showed exclusively the presence of S-linalool in all the crude oils and in the distilled samples.  相似文献   
53.
This paper describes the demonstration of Leishmania donovani amastigotes in canine tissues by immunoperoxidase staining. An indirect immunoperoxidase method was applied to the organs of 20 dogs in which leishmaniasis was previously diagnosed. Haemosiderin pigment was eliminated with 5 per cent oxalic acid. Amastigotes of L donovani appeared as dark brown stained bodies which contrasted with haematoxylin stained host cells. No positively stained amastigotes could be seen in any of the sections incubated with control serum. The organs which more frequently showed leishmanids were: skin (macrophages and fibroblasts), liver, spleen, lymph nodes and bone marrow. In a few cases amastigotes were seen in kidneys, gut, adrenal glands, eyes and testicles. This technique is simple to perform, gives consistent results and allows unequivocal histopathological diagnosis of canine leishmaniasis.  相似文献   
54.
In six field experiments conducted over 2 years, the effect of one or two cuttings on grain yield of triticale (× Triticosecale Wittmack) were investigated. In addition, the relationships between triticale grain yield its components were analysed.
Grain yield of the uncut plots invariably exceeded the plots with one or two cuttings, regardless of the environment or year. The reductions in grain yield caused by one cutting ranged from 27 % to 60 % under rainfed conditions and 18 % to 20 % under irrigation. With two cuttings the reduction was 45 % to 70 % for rainfed, and 35 % to 48 % for irrigated conditions.
The yield components which most influenced grain yield of triticale under cutting treatment were the number of ears m−2 in all six experiments and the number of grains per ear under rainfed conditions. At the rainfed sites in 1989 cuttings diminished the number of ears m−2 mainly by increasing plant mortality. Nevertheless, at the rainfed sites of 1990 and at the irrigated site of both years, the reduction in the number of ears m−2 with cuttings was due principally to a lower survival of tillers per plant.  相似文献   
55.
In March of 1991, a disease that affected pregnant sows and caused a high mortality in unweaned piglets was detected in Spain. Based on the clinical signs observed, mystery swine disease, which had been described recently in Germany, Holland and Belgium, was suspected. From the samples obtained from the affected farm, a filtrable agent (0.22 micron) was isolated on cell culture. It produced cytopathic effects, its replication was intracytoplasmic, it was sensitive to chloroform, and cross-reacted with a Lelystad reference serum. When inoculated into pregnant sows, the agent produced inappetence for 2-4 days, without hyperthermia. One of the sows aborted at 100 days of gestation; the two others had delayed parturitions (days 115 and 116). There was a mixture of healthy piglets, mummified fetuses, stillbirths and weak piglets. Microscopic examination of the lungs of healthy piglets killed at 8 and 12 days of life revealed the presence of interstitial pneumonia. The sera from the three sows at 39 days after infection cross-reacted with the Lelystad virus (titres > or = 1/640), whereas pre-inoculation sera did not recognize it (titres < or = 1/10). This is the first report from Spain of the isolation of an agent (antigenically related to the Lelystad virus), capable of reproducing the disease previously designated as mystery swine disease.  相似文献   
56.
57.
In order to determine the occurrence, serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) strains, 153 fecal samples of cattle randomly selected from six dairy farms in Sao Paulo State, Brazil, were examined for Shiga toxin (Stx) production by the Vero cell assay. Feces were directly streaked onto MacConkey Sorbitol Agar and incubated at 37 degrees C overnight. Sorbitol-negative colonies (maximum 20) and up to 10 sorbitol-positive colonies from each plate were subcultured onto presumptive diagnostic medium IAL. Sorbitol-negative isolates were screened with O157 antiserum for identification of O157:H7 E. coli. Isolates presenting cytotoxic activity were submitted to colony hybridization assays with specific DNA probes for stx1, stx2, eae, Ehly and astA genes. The isolation rate of STEC ranged from 3.8 to 84.6% depending on the farm analysed. STEC was identified in 25.5% of the animals, and most of them (64.1%) carried a single STEC serotype. A total of 202 STEC isolates were recovered from the animals, and except for the 2 O157:H7 isolates all the others expressed cytotoxic activity. The great majority of the STEC isolates carried both stx1 and stx2 genes (114/202, 56.4%) or stx2 (82/202, 40.6%); and whereas the Ehly sequence occurred in most of them (88%) eae was only observed in O157:H7 and O111:HNM isolates. Serotypes O113:H21, O178:H19 and O79:H14 were the most frequent STEC serotypes identified and widely distributed among animals from different farms, while others such as O77:H18, O88:H25 and O98:H17 occurred only in particular farms. This is the first report on the occurrence of STEC in dairy cattle in Sao Paulo State, and the results point to substantial differences in rate of isolation, serotypes and genetic profile of STEC that has been previously described among beef cattle in our community. Moreover, to our knowledge O79:H14 and O98:H17 represent new STEC serotypes, while O178:H19 has only been recently reported in Spain.  相似文献   
58.
Knowledge of spatial variability of soil fertility and plant nutrition is critical for planning and implementing site-specific vineyard management. To better understand the key drivers behind vineyard variability, yield mapping from 2002 to 2005 and 2007 (the monitor broke down in 2006) was used to identify zones of different productive potential in a Pinot Noir field located in Raimat (Lleida, Spain). Simultaneously, the vineyard field was sampled in 2002, 2003 and 2007, applying three different schemes (depending on the number of target vines in different grape yield zones). The sampling carried out in 2002, which involved different soil, topographic and crop properties (mineral contents in petiole), made it possible to evaluate the influence of these parameters on the grape yield variability. The zones of lowest yield coincided with locations in which the nutritional status of the crop exhibited the lowest values, particularly with respect to petiole contents of calcium and manganese. Sampling systems adopted in 2003 and 2007 (grape quality and soil attributes) confirmed the inverse spatial correlation between grape yield and some grape quality parameters and, more importantly, showed that the percentage of soil carbonates had a great influence on grape quality probably due to the reduced availability of manganese in calcareous soils. Site-specific vineyard management could therefore be considered using two different strategies: variable-rate application of foliar fertilizers to increase the yield in areas with low production and also foliar or soil fertilizers to improve the quality specifications in some areas.  相似文献   
59.
60.
This study aimed to evaluate the effects of a constant rate infusion (CRI) of xylazine or xylazine in combination with lidocaine on nociception, sedation, and physiologic values in horses. Six horses were given intravenous (IV) administration of a loading dose (LD) of 0.55 mg/kg of xylazine followed by a CRI of 1.1 mg/kg/hr. The horses were randomly assigned to receive three treatments, on different occasions, administered 10 minutes after initiation of the xylazine CRI, as follows: control, physiologic saline; lidocaine low CRI (LLCRI), lidocaine (LD: 1.3 mg/kg, CRI: 0.025 mg/kg/min); and lidocaine high CRI (LHCRI), lidocaine (LD: 1.3 mg/kg, CRI: 0.05 mg/kg/min). A blinded observer assessed objective and subjective data for 50 minutes during the CRIs. In all treatments, heart and respiratory rates decreased, end-tidal carbon dioxide concentration increased, and moderate to intense sedation was observed, but no significant treatment effect was detected in these variables. Ataxia was significantly higher in LHCRI than in the control treatment at 20 minutes of infusion. Compared with baseline values, nociceptive threshold increased to as much as 79% in the control, 190% in LLCRI, and 158% in LHCRI. Nociceptive threshold was significantly higher in LLCRI (at 10 and 50 minutes) and in LHCRI (at 30 minutes) than in the control treatment. The combination of CRIs of lidocaine with xylazine produced greater increases in nociceptive threshold compared with xylazine alone. The effects of xylazine on sedation and cardiorespiratory variables were not enhanced by the coadministration of lidocaine. The potential to increase ataxia may contraindicate the clinical use of LHCRI, in combination with xylazine, in standing horses.  相似文献   
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