Using homology modeling to interrogate binding affinity in neutralization of ricin toxin by a family of single domain antibodies

In this report we investigated, within a group of closely related single domain camelid antibodies (V_HHs), the relationship between binding affinity and neutralizing activity as it pertains to ricin, a fast‐acting toxin and biothreat agent. The V1C7‐like V_HHs (V1C7, V2B9, V2E8, and V5C1) are similar in amino acid sequence, but differ in their binding affinities and toxin‐neutralizing activities. Using the X‐ray crystal structure of V1C7 in complex with ricin's enzymatic subunit (RTA) as a template, Rosetta‐based homology modeling coupled with energetic decomposition led us to predict that a single pairwise interaction between Arg29 on V5C1 and Glu67 on RTA was responsible for the difference in ricin toxin binding affinity between V1C7, a weak neutralizer, and V5C1, a moderate neutralizer. This prediction was borne out experimentally: substitution of Arg for Gly at position 29 enhanced V1C7's binding affinity for ricin, whereas the reverse (ie, Gly for Arg at position 29) diminished V5C1's binding affinity by >10 fold. As expected, the V5C1_R29G mutant was largely devoid of toxin‐neutralizing activity (TNA). However, the TNA of the V1C7_G29R mutant was not correspondingly improved, indicating that in the V1C7 family binding affinity alone does not account for differences in antibody function. V1C7 and V5C1, as well as their respective point mutants, recognized indistinguishable epitopes on RTA, at least at the level of sensitivity afforded by hydrogen‐deuterium mass spectrometry. The results of this study have implications for engineering therapeutic antibodies because they demonstrate that even subtle differences in epitope specificity can account for important differences in antibody function.     

Is There Division of Labor in Cooperative Pseudoscorpions? An Analysis of the Behavioral Repertoire of a Tropical Species

Division of labor is a strategy that maximizes the foraging and reproductive success of eusocial insects. Although some arachnids exhibit colony structure and social organization similar to that of hymenopterans, temporal polyethism has only been demonstrated in few species. The social organization of cooperative pseudoscorpions Paratemnoides nidificator is similar to that of social spiders, but it involves a clear division of labor. Work allocation was experimentally investigated in colonies composed of only one developmental stage (young or adults) or by one sex (males or females), through laboratory manipulation. During 44 h of observation, more than 14 000 behavioral repetitions were quantified, distributed in 95 different types of behavioral acts, and grouped in 10 behavioral categories. The results showed that reproductive colonies of P. nidificator are maintained by gender‐ and age‐based activities. Males and non‐reproductive females performed the external cleaning of the colony and prey capture. Reproductive females take care of the juveniles and build reproductive silk chambers. Nymphs build most of the molt chambers and perform internal cleaning. In the absence of nymphs, male colonies survived 1–2 mo, while female colonies survived 3–4 mo. In nymph colonies, work is readjusted so that all maintenance tasks are executed. This is the first study clearly demonstrating division of tasks in arachnids. It suggests that specialization is an adaptative and evolutionarily old trait in this species. Unlike cooperative spiders, P. nidificator possesses physiological (e.g. reproduction, ecdysis, lifespan) and behavioral (e.g. behavioral synchrony or self‐organization) characteristics that allow task specialization.     

Length–weight relationships of 26 fish species from the middle section of the Negro River (Tacuarembó‐Durazno,Uruguay)

This study provides data on the length–weight relationship (LWR) for 26 species of neotropical fishes. Specimens were collected between 2008 and 2009 from a dam‐enclosed section of the middle Negro River (Uruguay). This study represents the first reference on length–weight relationships for 17 species, and also provides new maximum sizes for eight species.     

Prevalence of potentially pathogenic culturable bacteria on eggshells and in cloacae of female Pied Flycatchers in a temperate habitat in central Spain

ABSTRACT Bacteria grow on avian eggshells and thus can potentially cause diseases in developing embryos. Little is known about culturable bacteria colonizing avian eggshells in free‐living birds, with most studies restricted to poultry. Our objective was to examine the culturable bacterial array growing on eggshells during incubation that could negatively affect hatching success of Pied Flycatchers (Ficedula hypoleuca) in a temperate montane habitat in central Spain. Cloacal culturable bacteria of females were also analyzed because bacteria can be vertically transmitted from females to eggs. We used fecal samples as surrogates of cloacal samples due to the small size of sampled birds. We found that eggshells and female cloacae of Pied Flycatchers harbored 24 and 40 bacterial families and species, respectively, but only a few in each clutch and each cloaca. Rod‐shaped gram‐negative bacteria and bacteria in the family Pseudomonadaceae were the most common bacteria on eggshells during early and late incubation and in female cloacae. Although based on small sample sizes, we found that females with rod‐shaped gram‐negative bacteria in their cloacae laid eggs that also had these bacteria, providing possible evidence for vertical transmission. We found no evidence for vertical transmission of Pseudomonadaceae, suggesting a possible environmental source for these bacteria. The prevalence of bacterial morphological types and major taxonomical categories on eggshells did not vary from early to late stages of incubation, providing support for the hypothesis that incubation may have bacteriostatic effects on bacterial proliferation on eggshells. Despite being primary egg invaders in poultry, we detected no effects of culturable Pseudomonadaceae or Pseudomonas luteola on hatching success. Our study represents the first to examine the culturable bacteria growing on the eggshells of a wild bird in a temperate habitat and additional studies based on culture‐independent techniques are required to confirm our results.     

Systematics and cladistics of Megalostomis Chevrolat,and the biogeography of Clytrini (Coleoptera: Cryptocephalinae)

We present a cladistic analysis of the subtribe Megalostomina, a Neotropical group of ‘case‐bearer’ leaf beetles. A comparative study of the external and internal adult morphology of Clytrini was undertaken. New characters are described for the subtribe Megalostomina, from the internal sac of aedeagus, which provide a useful phylogenetic signal. More than 180 photographs illustrating the most important characters (74 characters and their respective states) used in the cladistic analysis are provided. The cladistic analysis of 57 terminal taxa and 95 characters was undertaken, under equal weights, and also using implied weights as a means to down‐weight homoplasious characters. We test the monophyly and explore intergeneric relationships of the subtribe Megalostomina, and reconstruct the relationships among the species of Megalostomis Chevrolat. The 42 species recognized can be assigned either to a group mostly containing species of North and Central America, or to a larger one of mostly South American species. Support is low, and the formal naming of groups is deferred pending a revision of all Megalostomina. We confirm the subgenera of Megalostomis of previous classifications are unnatural, and the following changes in the generic classification of the subtribe Megalostomina are proposed: Coleorozena Moldenke syn.n. of Coscinoptera Lacordaire; Coleothorpa Moldenke syn.n. of Coscinoptera Lacordaire; and Euryscopa (Coleoguerina) Moldenke syn.n. of Coscinoptera Lacordaire. Furthermore, six formerly recognized subgenera of Megalostomis are considered junior synonyms of Megalostomis Chevrolat: Megalostomis (Minturnia) Lacordaire syn.n. ; Megalostomis (Heterostomis) Lacordaire syn.n. ; Megalostomis (Scaphigenia) Lacordaire syn.n. ; Megalostomis (Snellingia) Moldenke syn.n. ; Megalostomis (Coleobyersa) Moldenke syn.n. ; and Megalostomis (Pygidiocarina) Moldenke syn.n. Thus, no subgenera are recognized within Megalostomis. Previous hypotheses on Clytrini biogeography were revisited in the light of new biogeographic and phylogenetic knowledge. We hypothesize an origin of Clytrini in tropical/subtropical Gondwana, when South America, Africa, Madagascar and India were connected. Changes in the configuration of the tectonic plates in the Cenozoic allowed the dispersal of Clytrina to the Palaearctic and Nearctic regions, and dispersion of Babiina and Megalostomina through the Nearctic region.