Layout proposal for a microtitre plate to be used in routine antimicrobial susceptibility testing of bacteria from infections of dogs and cats

The determination of minimum inhibitory concentrations by broth microdilution is recommended as method of choice for susceptibility testing of veterinary bacterial pathogens. Accordingly, broth microdilution is used in veterinary routine diagnostic laboratories at a progressive rate. To reduce the costs of susceptibility testing, it is reasonable to develop widely accepted uniform microtitre plate layouts that are produced in large quantities. Such microtitre plate layouts have already been developed and published for the susceptibility testing of pathogens from food-producing animals. However, a microtitre plate layout, especially designed for the testing of bacteria from dogs and cats, should be available, too. The choice of the antimicrobial agents or combinations of antimicrobial agents to be included in a suitable layout should be based on the following criteria: (1) the approval and availability of an antimicrobial agent or combination of agents, (2) known cross-resistances, and (3) availability of approved clinical breakpoints. The latter point is of particular importance for the choice of the numbers of concentrations per antimicrobial agent tested and the range of test concentrations. Taking into account these aspects, a science-based layout proposal for microtitre plates, which are suitable for routine testing of bacteria from dogs and cats, is presented and discussed.     

Uncertain water supply in an irrigated Mediterranean area: An analysis of the possible economic impact of climate change on the farm sector

Analysis of the possible economic impact of climate change at the local level is becoming increasingly relevant to agricultural policy, in terms of the definition of new measures to sustain adaptation of the farm sector. This study focuses on a Mediterranean agricultural zone to evaluate the economic impact of rainfall regime changes that modify the accumulation of irrigation water in a dam. The objective is to identify farm typologies that suffer more from rainfall changes, in order to target policy measures that increase farm sector capability to adapt to climate change. First, an analysis of historical series is conducted for precipitation. The decreasing trend in annual precipitation, as well as an increase in monthly rainfall variability, is shown to have a statistically significant influence on the regime of water accumulation in the dam. Density functions representing this regime are estimated for several periods, including the 1960s-1970s, the current time and a time interval that extends to 2015. A comparison of these functions reveals an increase in variability of water accumulation in the dam through time. Parameters of these functions are used in three models of Discrete Stochastic Programming to represent different expectations of irrigation water availability and to simulate the possible reaction of the farm sector in the study area to the different scenarios. The simulation results show that both income and employment are noticeably reduced in some farm typologies when scenarios with higher variability levels for water accumulation in the dam are considered. In addition, changes in the use of soil are seen, the use of inputs declines and the quantity of extracted groundwater increases.     

Control of spawning activity in female Nile tilapia (Oreochromis niloticus) (L.) by temperature manipulation

The aim of this study was to maximize the spawning of Oreochromis niloticus females in a specific time period. Females were divided randomly into control and treatment groups. In the treatment groups, females were kept for one week at 28±0.5 °C, after which they were exposed to a reduced water temperature of 22±0.5 °C for 7, 14 and 28 days. Thereafter, the temperature was restored to 28 °C. Females in the control groups were kept continuously at a water temperature of 28 °C. All females were checked daily for signs of spawning for the duration of the experiments and were manually stripped if ready to spawn. The following parameters were calculated for period of 3 and 7 days following a 28 °C temperature restoration: spawning rate, number of eggs per female, weight of female, relative fecundity (eggs g⁻¹ body weight) and the percentage of hatched and swim-up fry. The highest spawning rate of 39.5% was obtained in the 14-day trial over a period of 7 days, while the corresponding value in the control was 12.5%. The percentages of hatched and swim-up fry in the 14- and 28-day trials, however, were significantly higher in the controls than in the corresponding treatment groups.     

Design of Inhibitors of Photosystem II using a Model of the D1 Protein

Several inhibitors belonging to structurally different chemical classes were used to analyze the predictive power of an initial model of the herbicide binding niche of the D1 protein belonging to photosystem II (PS II) from plants. In the case of small PS II inhibitors, the estimation of relative activities was hampered by uncertainty about the binding modes. To overcome this problem, a bulky substituent was introduced into the inhibitors to act as a hook, resulting in an unambiguous orientation in the model. The comparison of the modelling results and the experimentally determined IC₅₀ values of different triazines suggested that the previously assumed volume of the binding niche had to be reduced by 20%. After refinement of the model, it was possible to estimate qualitatively, the relative in-vitro activity for inhibitors belonging to different families, as long as an unambiguous binding mode could be deduced either from steric demands or from IC₅₀ values of mutant D1 proteins. The usefulness of the refined model is demonstrated by the successful de-novo design of a potent class of herbicides, the triazolopyrimidines.     

A genomic regulatory network for development

Development of the body plan is controlled by large networks of regulatory genes. A gene regulatory network that controls the specification of endoderm and mesoderm in the sea urchin embryo is summarized here. The network was derived from large-scale perturbation analyses, in combination with computational methodologies, genomic data, cis-regulatory analysis, and molecular embryology. The network contains over 40 genes at present, and each node can be directly verified at the DNA sequence level by cis-regulatory analysis. Its architecture reveals specific and general aspects of development, such as how given cells generate their ordained fates in the embryo and why the process moves inexorably forward in developmental time.