Diet of land birds along an elevational gradient in Papua New Guinea.

Food preferences and exploitation are crucial to many aspects of avian ecology and are of increasing importance as we progress in our understanding of community ecology. We studied birds and their feeding specialization in the Central Range of Papua New Guinea, at eight study sites along a complete (200 to 3700 m a.s.l.) rainforest elevational gradient. The relative species richness and abundance increased with increasing elevation for insect and nectar eating birds, and decreased with elevation for fruit feeding birds. Using emetic tartar, we coerced 999 individuals from 99 bird species to regurgitate their stomach contents and studied these food samples. The proportion of arthropods in food samples increased with increasing elevation at the expense of plant material. Body size of arthropods eaten by birds decreased with increasing elevation. This reflected the parallel elevational trend in the body size of arthropods available in the forest understory. Body size of insectivorous birds was significantly positively correlated with the body size of arthropods they ate. Coleoptera were the most exploited arthropods, followed by Araneae, Hymenoptera, and Lepidoptera. Selectivity indexes showed that most of the arthropod taxa were taken opportunistically, reflecting the spatial patterns in arthropod abundances to which the birds were exposed.

The role of key features in predator recognition by untrained birds.

The most important role in the recognition and categorization of predators (as well as other animals) is usually attributed to so-called key features. Under laboratory conditions, we tested the role of yellow eyes (specific for the genus Accipiter in European raptors) and hooked beak (common for all European birds of prey) in the recognition of the sparrowhawk (Accipiter nisus) by untrained great tits (Parus major) caught in the wild. Using wooden dummies, we interchanged either one of these potential key features or the body of the sparrowhawk (predator) and domestic pigeon (harmless bird). The tested tits showed three types of behaviour in the presence of the dummies: fear, interest without fear, and lack of interest. Eye interchange lowered fear of the sparrowhawk, but did not cause fear of the pigeon. Beak interchange did not lower fear of the sparrowhawk. Eye interchange caused increased interest in both species. Thus, a specific sparrowhawk feature is necessary for correct sparrowhawk dummy recognition but a general raptor feature is not. On the other hand, a specific sparrowhawk feature on a pigeon dummy is not enough to prompt sparrowhawk recognition. Thus, key features play an important, but not exclusive, role in predator recognition. An increased interest in some of the modified dummies implies that the tits have a general concept of a sparrowhawk. The individual variability in behaviour of tits is discussed.

Modification of activity and specificity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26 by engineering of its entrance tunnel.

Structural comparison of three different haloalkane dehalogenases suggested that substrate specificity of these bacterial enzymes could be significantly influenced by the size and shape of their entrance tunnels. The surface residue leucine 177 positioned at the tunnel opening of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 was selected for modification based on structural and phylogenetic analysis; the residue partially blocks the entrance tunnel, and it is the most variable pocket residue in haloalkane dehalogenase-like proteins with nine substitutions in 14 proteins. Mutant genes coding for proteins carrying all possible substitutions in position 177 were constructed by site-directed mutagenesis and heterologously expressed in Escherichia coli. In total, 15 active protein variants were obtained, suggesting a relatively high tolerance of the site for the introduction of mutations. Purified protein variants were kinetically characterized by determination of specific activities with 12 halogenated substrates and steady-state kinetic parameters with two substrates. The effect of mutation on the enzyme activities varied dramatically with the structure of the substrates, suggesting that extrapolation of one substrate to another may be misleading and that a systematic characterization of the protein variants with a number of substrates is essential. Multivariate analysis of activity data revealed that catalytic activity of mutant enzymes generally increased with the introduction of small and nonpolar amino acid in position 177. This result is consistent with the phylogenetic analysis showing that glycine and alanine are the most commonly occurring amino acids in this position among haloalkane dehalogenases. The study demonstrates the advantages of using rational engineering to develop enzymes with modified catalytic properties and substrate specificities. The strategy of using site-directed mutagenesis to modify a specific entrance tunnel residue identified by structural and phylogenetic analyses, rather than combinatorial screening, generated a high percentage of viable mutants.

Reconstruction of mycobacterial dehalogenase Rv2579 by cumulative mutagenesis of haloalkane dehalogenase LinB.

The homology model of protein Rv2579 from Mycobacterium tuberculosis H37Rv was compared with the crystal structure of haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26, and this analysis revealed that 6 of 19 amino acid residues which form an active site and entrance tunnel are different in LinB and Rv2579. To characterize the effect of replacement of these six amino acid residues, mutations were introduced cumulatively into the six amino acid residues of LinB. The sixfold mutant, which was supposed to have the active site of Rv2579, exhibited haloalkane dehalogenase activity with the haloalkanes tested, confirming that Rv2579 is a member of the haloalkane dehalogenase protein family.