Temporal Distribution and Weather Correlates of Norway Rat (Rattus norvegicus) Infestations in the City of Madrid, Spain

Urban Norway rats are challenging pests, posing significant health and economic threats. Implementing ecologically based integrated rodent management (EBIRM) programmes relies primarily on the understanding of ecological relationships between rodents and their environments, with emphasis on the processes influencing rodent populations in the target ecosystem. We investigated the temporal distribution of urban Norway rat infestations in Madrid, Spain, and tested for the association of such infestations with temperature, relative humidity and precipitation by fitting a multivariate Poisson generalized linear model to a 3-year (2006–2008) daily time series of 4,689 Norway rat sightings. Norway rat infestations showed a marked seasonality, peaking in the summer. Most Norway rat sightings were reported on Mondays. Minimum temperature and relative humidity were positively associated with Norway rat infestation, whereas the association with precipitation was negative. The time series was adequately explained by the model. We identified previously unrecognized time periods that are more prone to Norway rat infestation than others and generated hypotheses about the association between weather, human outdoor activity, resource availability, rodent activity and population size. This provided local authorities engaged in preserving urban ecosystem health with basic research information to predict future rodent outbreaks and support the implementation of EBIRM programmes in urban areas.     

Characterization of Growth-Retardant Effects on Vegetative Growth of Date Palm Seedlings

The leaves, inflorescences, and fruit bunches of date palm trees develop at the crown, which is the apex of a single trunk that can be up to 25 m tall. Because palm trees are monocots, the leaves and inflorescences are generated deep within the trunk from a single meristem—the palm heart—and are inaccessible for direct bioregulator treatments. We followed the effects of two growth regulators, paclobutrazol and uniconazole-P, on the vegetative growth of young date palm seedlings. Both growth regulators constrained the elongation rate of the leaves and reduced the leaf length. Microscopic analysis of dissected palm hearts revealed that the growth retardants did not affect the generation of new leaves. However, the treatments did change the dimensions of the developing embryonic leaves as well as the shape of the apical meristem. The results suggest the possible future use of growth retardants to reduce trunk height, enabling cheaper, safer, and more efficient management of date palm trees.     

Cultivation-independent approaches to investigate the chemistry of marine symbiotic bacteria

Sessile marine animals like sponges, tunicates, and bryozoans are a rich source of bioactive natural products, many of which exhibit potent anticancer activities. However, most of these substances are available in very limited amounts only, which has prohibited further drug development. Recent evidence suggests that symbiotic bacteria might be the true producers of many animal-derived metabolites. In addition to revealing fascinating perspectives for research in marine chemical ecology, these findings suggest new solutions to the supply problem. Although most symbionts remain uncultivated, bacterial production systems might be created by isolating biosynthetic genes from marine metagenomes, and expressing them in culturable bacterial hosts. This review discusses cell-sorting, natural product visualization, and phylogenetic approaches to identify symbiotic producers. In addition, strategies to isolate genes and gene clusters from marine species consortia are described. These techniques have provided insights into the bacterial origin and biosynthesis of polyketides like the onnamides, swinholides, and bryostatins, of peptides including the patellamides, chlorinated dipeptides, and theopalauamide as well as of brominated biphenylethers.     

Production and Characterization of a Thermostable Alcohol Dehydrogenase That Belongs to the Aldo-Keto Reductase Superfamily

The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.     

Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily

The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100 degrees C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.     

Directed evolution of sulfotransferases and paraoxonases by ancestral libraries

Large libraries of randomly mutated genes are applied in directed evolution experiments in order to obtain sufficient variability. These libraries, however, contain mostly inactive variants, and the very low frequency of improved variants can only be isolated by high-throughput screening. Small but efficient libraries comprise an attractive alternative. Here, we describe the application of ancestral libraries—libraries based on mutations predicted by phylogenetic analysis and ancestral inference. We designed and constructed such libraries using serum paraoxonases and cytosolic sulfotransferases (SULTs) as model enzymes. Both of these enzyme families exhibit a range of activities in drug metabolism and detoxification of xenobiotics. The ancestral serum paraoxonase and SULT libraries were screened by low-throughput means, including HPLC, using substrates and/or reactions with which all family members exhibit low activity. The libraries showed a remarkably high frequency of highly polymorphic and functionally diverse variants. Screening of as few as 300 variants enabled the isolation of mutants with up to 50-fold higher activity than the starting point enzyme. Structural and kinetic characterizations of an evolved SULT variant show how few ancestral mutations reshaped the active site and modulated the enzyme's specificity. Ancestral libraries therefore comprise a means of focusing diversity to positions and mutations that readily trigger changes in substrate and/or reaction specificity, thereby facilitating the isolation of new enzyme variants for a variety of different substrates and reactions by medium-throughput or even low-throughput screens.     

A small fishery with a high impact on sea turtle populations in the eastern Mediterranean

Sea turtles were targeted by fisheries in the Mediterranean from 1920 to 1970 and have undergone severe exploitation. At least 30,000 to 40,000 turtles were caught along the Palestinian coastline during the 1920s to 1930s. Although intentional cap- ture of marine turtles is now illegal, sea turtles are still incidentally caught by the fishing industry, making it a major cause of sea turtle mortality. The present study as- sesses the impact of the Israeli fishery fleet on the turtle population in the Levantine basin based on on-board observations and a fishermen survey. The results show that gillnets and trawlers are the main threats to sea turtles in this area. 21 turtles were caught during 1385.5 hours of trawling observations – a catch rate of 0.015 turtles per hour. We estimate that a total of 1,315 turtles are caught annually by Israeli trawlers. According to the fishermen survey, ～21 turtles are caught each year by a single gillnet vessel, yielding an annual estimate of 1,672 turtles for the whole gillnet fleet. We have also found that only a small fraction of the turtles injured by trawlers is represented in the strandings. The mortality rate through trawling and the stranding density is the highest in the region, emphasizing the urgent need to regulate the Israe- li fishery. This fishery poses a major threat to the whole Levantine sea turtle popula- tion, especially during the vulnerable reproduction stage.