Selection and characterization of a proteo-chitinolytic strain of Bacillus thuringiensis,able to grow in shrimp waste media

This paper reports the selection and characterization of Bacillus thuringiensis strains, with ability to grow in a proteo-chitinaceous substrate (milled shrimp waste) as the sole ingredient. Selected strains were able to produce crystal proteins, as well as proteases and chitinases as fermentation by-products. By a preliminary, qualitative screening of 152 B. thuringiensis strains, grown on media rich in protein and chitin, eight strains were selected. These strains were cultured in a liquid medium containing milled shrimp waste and their kinetics of protease production were followed. The two most active proteolytic strains (Bt-103 and Bt-112) were characterized by their crystal protein content, plasmid profiles, crystal ultrastructure, and toxicity towards Manduca sexta, Aedes aegypti and Leptinotarsa texana. The only activity recorded in these species was moderate toxicity of strain Bt-112 against Manduca sexta first instar larvae, as well as the highest proteolytic and chitinolytic activities. Its bipyramidal crystals were associated with semi-cuboidal inclusions and although its crystal proteins were similar to those of B. thuringiensis kurstaki (HD-1), its plasmid content was quite different. Serotyping of Bt-112 indicated that it belongs to serovar. tolworthi. Further studies with a similar strategy might render more strains with ability to grow in a rich waste by-product like the shrimp waste, which may show not only higher insecticidal activity, but also with the ability to produce extracellular enzymes with biotechnological applications.     

The conservation value of tree decay processes as a key driver structuring tree cavity nest webs in South American temperate rainforests

South American temperate rainforests, a global biodiversity hotspot, have been reduced to nearly 30% of their original extent and most remaining stands are being degraded. Cavity-nesting vertebrate communities are dependent on cavity-bearing trees and hierarchically structured within nest webs. Evaluating the actual degree of cavity dependence (obligate, non-obligate) and the preferred attributes of trees by cavity nesters is critical to design conservation strategies in areas undergoing habitat loss. During three breeding seasons (2010–2013), we studied the cavity-nesting bird community in temperate rainforests of Chile. We found the highest reported proportion of tree cavity nesters (n = 29 species; 57%) compared to non-cavity-using birds for any forest system. Four species were excavators and 25 were secondary cavity nesters (SCNs). Among SCNs, ten species were obligate and 15 were non-obligate cavity nesters. Seventy-five percent of nests of SCNs were located in cavities produced by tree decay processes and the remaining 25% were in cavities excavated mainly by Pygarrhichas albogularis and Campephilus magellanicus. Nest web structure had a low dominance and evenness, with most network interactions occurring between SCNs and large decaying trees. Tree diameter at breast height (DBH) was larger in nest-trees (57.3 cm) than in available trees (26.1 cm). Cavity nesters showed a strong preference for dead trees, both standing and fallen (58% of nests). Our results stress that retaining large decaying and standing dead trees (DBH > 57 cm), and large fallen trees, should be a priority for retention in forest management plans in this globally threatened ecosystem.     

A new family of H3 receptor antagonists based on the natural product Conessine

A new family of Histamine H(3) receptor antagonists (5a-t) has been prepared based on the structure of the natural product Conessine, a known H(3) antagonist. Several members of the new series are highly potent and selective binders of rat and human H(3) receptors and display inverse agonism at the human H(3) receptor. Compound 5n exhibited promising rat pharmacokinetic properties and demonstrated functional antagonism of the H(3) receptor in an in-vivo pharmacological model.     

Genotype by environment interaction for adult body weights of shrimp Penaeus vannamei when grown at low and high densitie

Shrimp is one of few marine species cultured worldwide for which several selective breeding programs are being conducted. One environmental factor that can affect the response to selection in breeding programs is the density at which the shrimp are cultured (low-medium-high). Phenotypic plasticity in the growth response to different densities might be accompanied by a significant genotype by environment interaction, evidenced by a change in heritabilities between environments and by a genetic correlation less than one for a unique trait between environments. Our goal was to understand whether different growth densities affect estimates of those genetic parameters for adult body weight (BW) in the Pacific white shrimp (Penaeus vannamei). BW heritabilities were significantly different between environments, with the largest at high density. These differences resulted from both an increased additive genetic variance and a decreased environmental variance when grown at high density. The genetic correlation between BWs at the two environmental conditions was significantly less than one. Whereas these results might be suggestive for carrying out shrimp selective breeding for BW under high density conditions, further understanding of genetic correlations between growth and reproductive traits within a given environment is necessary, as there are indications of reduced reproductive fitness for shrimp grown at high densities.     

Evaluation of immune responses raised against Tc13 antigens of Trypanosoma cruzi in the outcome of murine experimental infection

We have previously reported that genetic immunization with Tc13Tul antigen of Trypanosoma cruzi, the aetiological agent of Chagas' disease, triggers harmful effects and non-protective immune responses. In order to confirm the role of Tc13 antigens during T. cruzi infection, herein we studied the humoral and cellular immune responses to the Tc13Tul molecule and its EPKSA C-terminal portion in BALB/c T. cruzi-infected mice or mice immunized with recombinant Tc13Tul. Analysis of the antibody response showed that B-cell epitopes that stimulate a sustained IgM production along the infection and high levels of IgG in the acute phase are mainly located at the Tc13 N- and C-terminal domains, respectively. DTH assays showed that T-cell epitopes are mainly at the Tc13 N-terminal segment and that they do not elicit an efficient memory response. Recombinant Tc13Tul did not induce IFN-gamma secretion in either infected or immunized mice. However, a putative CD8+Tc13Tul-derived peptide was found to elicit IFN-gamma production in chronically infected animals. Immunization with recombinant Tc13Tul did not induce pathology in tissues and neither did it protect against the infection. Our results show that in the outcome of T. cruzi infection the Tc13 family protein mainly triggers non-protective immune responses.     

Induction of immunity in sheep to Fasciola hepatica with mimotopes of cathepsin L selected from a phage display library

An M13 phage random 12-mers peptide library was used to screen cathepsin L mimotopes of Fasciola hepatica and to evaluate their immunogenicity in sheep. Seven clones showed positive reactivity to a rabbit anti-cathepsin L1/L2 antiserum in ELISA, and their amino acid sequences deduced by DNA sequencing were tentatively mapped on the protein. Twenty sheep were randomly allocated into 4 groups of 5 animals each, for immunization with 1x10(14) phage particles of clones 1, 20, a mixture of 7 clones and PBS, without adjuvant at the beginning, and 4 weeks later. All groups were challenged with 300 metacercariae at week 6 and slaughtered 16 weeks later. The mean worm burdens after challenge were reduced by 47.61% and 33.91% in sheep vaccinated with clones 1 and 20, respectively; no effect was observed in animals inoculated with the clone mixture. Also, a significant reduction in worm size and burden was observed for those sheep immunized with clone 1. Animals receiving clone 20, showed a significant reduction in egg output. Immunization induced a reduction of egg viability ranging from 58.92 to 82.11%. Furthermore, vaccinated animals produced clone-specific antibodies which were boosted after challenge with metacercariae of F. hepatica.     

What can be learnt from analysing insect orientation flights using probabilistic SLAM?

In this paper, we provide an analysis of orientation flights in bumblebees, employing a novel technique based on simultaneous localisation and mapping (SLAM) a probabilistic approach from autonomous robotics. We use SLAM to determine what bumblebees might learn about the locations of objects in the world through the arcing behaviours that are typical of these flights. Our results indicate that while the bees are clearly influenced by the presence of a conspicuous landmark, there is little evidence that they structure their flights to specifically learn about the position of the landmark.     

In vitro regeneration of the important North American oak species <Emphasis Type="Italic">Quercus alba</Emphasis>, <Emphasis Type="Italic">Quercus bicolor</Emphasis> and <Emphasis Type="Italic">Quercus rubra</Emphasis>

North American oak species, with their characteristic strong episodic seasonal shoot growth, are highly problematic for clonal micropropagation, resulting in the inability to achieve a stabilized shoot multiplication stage. The potential for initiating and proliferating shoot cultures derived from Quercus alba, Q. bicolor and Q. rubra explants was investigated, and a micropropagation method for these species was developed. Branch segments from 6 to 7-year-old trees were forced-flushed and the forced shoots were used as source of explants for culture initiation. A consistent shoot multiplication stage was achieved, in 13 of the 15 genotypes established in vitro, although marked differences occurred in explants from different genotypes/species. The control of efficient shoot multiplication involved the culture of decapitated shoots in a stressful horizontal position on cytokinin-containing medium with a sequence of transfers within a 6-week subculture cycle, which was beneficial to overcoming the episodic character of shoot growth. During each subculture cycle, the horizontally placed explants were cultured on media containing 0.2 mg l⁻¹ benzyladenine (BA) for 2 weeks with two successive transfers (2 weeks each) to fresh medium with 0.1 mg l⁻¹ BA, giving a 6-week subculture cycle. The general appearance and vigor of Q. alba and Q. bicolor shoot cultures were improved by the inclusion of both 0.1 mg l⁻¹ BA and 0.5 mg l⁻¹ zeatin in the medium used for the second transfer within the 6-week subculture cycle. Addition of AgNO₃ (3 mg l⁻¹) to the shoot proliferation medium of Q. rubra had a significant positive effect on shoot development pattern by reducing deleterious symptoms, including shoot tip necrosis and early senescence of leaves. The three species showed acceptable in vitro rooting rates by culturing microcuttings in medium containing 25 mg l⁻¹ indolebutyric acid for 48 h with subsequent transfer to auxin-free medium supplemented with 0.4% activated charcoal. Although an initial 5-day dark period generally improved the rooting response, it was detrimental to the quality of regenerated plantlets. However, activated charcoal stimulated not only the rooting frequencies, but it also enhanced plant quality, as evidenced by root, shoot and leaf growth.     

Isolation, characterization and cross-amplification of polymorphic microsatellite loci in the threatened endemic Mammillaria crucigera (Cactaceae)

The cactus Mammillaria crucigera is a threatened species endemic to central Mexico. As a means of assessing population genetic status of these species, eight microsatellite markers were developed. These primers were tested in 40 individual from two wild populations. The results showed that these primers will be useful to describe population structure and aid to the conservation of species. The eight primers were tested in other Mammillaria species and most of them showed successful amplification.     

Nap1 regulates Dictyostelium cell motility and adhesion through SCAR-dependent and -independent pathways

SCAR--also known as WAVE--is a key regulator of actin dynamics. Activation of SCAR enhances the nucleation of new actin filaments through the Arp2/3 complex, causing a localized increase in the rate of actin polymerization . In vivo, SCAR is held in a large regulatory complex, which includes PIR121 and Nap1 proteins, whose precise role is unclear. It was initially thought to hold SCAR inactive until needed , but recent data suggest that it is essential for SCAR function . Here, we show that disruption of the gene that encodes Nap1 (napA) causes loss of SCAR function. Cells lacking Nap1 are small and rounded, with diminished actin polymerization and small pseudopods. Furthermore, several aspects of the napA phenotype are more severe than those evoked by the absence of SCAR alone. In particular, napA mutants have defects in cell-substrate adhesion and multicellular development. Despite these defects, napA(-) cells move and chemotax surprisingly effectively. Our results show that the members of the complex have unexpectedly diverse biological roles.