Use of marker genes in competition studies of Rhizobium

Use of marker genes has several advantages in studying rhizobial competition compared to traditional approaches. Reporter genes such as the ß-glucuronidase gene (gusA) or a thermostable ß-glucosidase gene (celB) allow detection of rhizobial strains in nodules when they are still attached to the root system. Analysis is extremely simple, fast and permits a high data throughput. This detection technique is therefore highly suitable for the study of rhizobial competition and studies using gusA-marked strains of Rhizobium are presented. By making use of gusA and celB, differentially marked strains can be produced and distinguished easily on roots. The availability of two marker genes permits competition studies of two or more than two strains and analysis of dual nodule occupancy. As this methodology does not require sophisticated equipment, a GUS Gene Marking Kit was developed.     

Enrofloxacin and Macrolides Alone or in Combination with Rifampicin as Antimicrobial Treatment in a Bovine Model of Acute Chlamydia psittaci Infection

Chlamydia psittaci is a zoonotic bacterium with a wide host range that can cause respiratory disease in humans and cattle. In the present study, effects of treatment with macrolides and quinolones applied alone or in combination with rifampicin were tested in a previously established bovine model of respiratory C. psittaci infection. Fifty animals were inoculated intrabronchially at the age of 6–8 weeks. Seven served as untreated controls, the others were assigned to seven treatment groups: (i) rifampicin, (ii) enrofloxacin, (iii) enrofloxacin + rifampicin, (iv) azithromycin, (v) azithromycin + rifampicin, (vi) erythromycin, and (vii) erythromycin + rifampicin. Treatment started 30 hours after inoculation and continued until 14 days after inoculation (dpi), when all animals were necropsied. The infection was successful in all animals and sufficient antibiotic levels were detected in blood plasma and tissue of the treated animals. Reisolation of the pathogen was achieved more often from untreated animals than from other groups. Nevertheless, pathogen detection by PCR was possible to the same extent in all animals and there were no significant differences between treated and untreated animals in terms of local (i.e. cell count and differentiation of BALF-cells) and systemic inflammation (i.e. white blood cells and concentration of acute phase protein LBP), clinical signs, and pathological findings at necropsy. Regardless of the reduced reisolation rate in treated animals, the treatment of experimentally induced respiratory C. psittaci infection with enrofloxacin, azithromycin or erythromycin alone or in combination with rifampicin was without obvious benefit for the host, since no significant differences in clinical and pathological findings or inflammatory parameters were detected and all animals recovered clinically within two weeks.     

The Role of China in the Global Spread of the Current Cholera Pandemic

Epidemics and pandemics of cholera, a severe diarrheal disease, have occurred since the early 19th century and waves of epidemic disease continue today. Cholera epidemics are caused by individual, genetically monomorphic lineages of Vibrio cholerae: the ongoing seventh pandemic, which has spread globally since 1961, is associated with lineage L2 of biotype El Tor. Previous genomic studies of the epidemiology of the seventh pandemic identified three successive sub-lineages within L2, designated waves 1 to 3, which spread globally from the Bay of Bengal on multiple occasions. However, these studies did not include samples from China, which also experienced multiple epidemics of cholera in recent decades. We sequenced the genomes of 71 strains isolated in China between 1961 and 2010, as well as eight from other sources, and compared them with 181 published genomes. The results indicated that outbreaks in China between 1960 and 1990 were associated with wave 1 whereas later outbreaks were associated with wave 2. However, the previously defined waves overlapped temporally, and are an inadequate representation of the shape of the global genealogy. We therefore suggest replacing them by a series of tightly delineated clades. Between 1960 and 1990 multiple such clades were imported into China, underwent further microevolution there and then spread to other countries. China was thus both a sink and source during the pandemic spread of V. cholerae, and needs to be included in reconstructions of the global patterns of spread of cholera.     

Investigation of Water Dynamics and the Effect of Evapotranspiration on Grain Yield of Rainfed Wheat and Barley under a Mediterranean Environment: A Modelling Approach

Agro-hydrological models have increasingly become useful and powerful tools in optimizing water and fertilizer application, and in studying the environmental consequences. Accurate prediction of water dynamics in such models is essential for models to produce reasonable results. In this study, detailed simulations were performed for water dynamics of rainfed winter wheat and barley grown under a Mediterranean climate over a 10-year period. The model employed (Yang et al., 2009. J. Hydrol., 370, 177-190) uses easily available agronomic data, and takes into consideration of all key soil and plant processes in controlling water dynamics in the soil-crop system, including the dynamics of root growth. The water requirement for crop growth was calculated according to the FAO56, and the soil hydraulic properties were estimated using peto-transfer functions (PTFs) based on soil physical properties and soil organic matter content. Results show that the simulated values of soil water content at the depths of 15, 45 and 75 cm agreed with the measurements well with the root of the mean squared errors of 0.027 cm³ cm^-3 and the model agreement index of 0.875. The simulated seasonal evapotranspiration (ET) ranged from 208 to 388 mm, and grain yield was found to correlate with the simulated seasonal ET in a linear manner within the studied ET range. The simulated rates of grain yield increase were 17.3 and 23.7 kg ha^-l for every mm of water evapotranspired for wheat and barley, respectively. The good agreement of soil water content between measurement and simulation and the simulated relationships between grain yield and seasonal ET supported by the data in the literature indicates that the model performed well in modelling water dynamics for the studied soil-crop system, and therefore has the potential to be applied reliably and widely in precision agriculture. Finally, a two-staged approach using inverse modelling techniques to further improve model performance was discussed.     

Natural history museum collections provide information on phenological change in British butterflies since the late-nineteenth century

Museum collections have the potential to provide valuable information on the phenological response of organisms to climate change. This is particularly useful for those species for which few data otherwise exist, but also to extend time series to the period before other observational data are available. To test this potential, we analysed data from 2,630 specimens of four species of British butterflies (Anthocharis cardamines, Hamearis lucina, Polyommatus bellargus and Pyrgus malvae), collected from 1876 to 1999 and stored in the Natural History Museum, London, UK (NHM). In A. cardamines, first-generation P. bellargus and P. malvae, we found that there was a strong significant negative relationship between spring temperature and 10th percentile collection dates, which approximates mean first appearance date, and median collection date, which approximates mean flight date. In all four species, there was a significant negative relationship between the 10th percentile collection date and the length of the collection period, which approximates flight period. In second-generation P. bellargus, these phenological measurements were correlated with summer temperature. We found that the rates of phenological response to temperature, based on NHM data, were similar to, or somewhat greater than, those reported for other organisms based on observational data covering the last 40 years. The lower rate of phenological response, and the significant influence of February rather than March or April temperatures, in recent decades compared with data from earlier in the twentieth century may indicate that early emerging British butterfly species are currently approaching the limits of phenological advancement in response to recent climate warming.