Abstract Background: Scorpion neurotoxins such as those that modify the mammalian voltage-gated sodium ion channels (Nav) are the main responsible for scorpion envenomation. Their neutralization is crucial in the production of antivenoms against scorpion stings. Methods: In the present study, two in silico designed genes - one that codes for a native neurotoxin from the venom of the Anatolian scorpion Androctonus crassicauda, named Acra 4 - and another non-native toxin - named consensus scorpion toxin (SccTx) obtained from the alignment of the primary structures of the most toxic neurotoxins from the Middle Eastern and North African scorpions - were recombinantly expressed in E. coli Origami. Results: Following bacterial expression, the two expressed neurotoxins, hereafter named HisrAcra4 and HisrSccTx, were obtained from inclusion bodies. Both recombinant neurotoxins were obtained in multiple Cys-Cys isoforms. After refolding, the active protein fractions were identified with molecular masses of 8,947.6 and 9,989.1 Da for HisrAcra4 and HisrSccTx, respectively, which agreed with their expected theoretical masses. HisrAcra4 and HisrSccTx were used as antigens to immunize two groups of rabbits, to produce either anti-HisrAcra4 or anti-HisrSccTx serum antibodies, which in turn could recognize and neutralize neurotoxins from venoms of scorpion species from the Middle East and North Africa. The antibodies obtained from rabbits neutralized the 3LD50 of Androctonus australis, Leiurus quinquestriatus hebraeus and Buthus occitanus venoms, but they did not neutralize A. crassicauda and A. mauritanicus venoms. In addition, the anti-HisrAcra4 antibodies did not neutralize any of the five scorpion venoms tested. However, an antibody blend of anti-HisrAcra4 and anti-HisrSccTx was able to neutralize A. crassicauda and A. mauritanicus venoms. Conclusions: Two recombinant Nav neurotoxins, from different peptide families, were used as antigens to generate IgGs for neutralizing scorpion venoms of species from the Middle East and North Africa.

Abstract Nectariferous and polliniferous resources are key to the survival of social bees, so identifying the origin sources allows the implementation of management actions aimed at a greater supply of nutrients for the colonies. Besides, defining the floral origin of honeys contributes to their characterization and commercialization. The objective of our work was to identify the plants that provide nectar to the bees Apis mellifera and Tetragonisca fiebrigi in northern Misiones, through palynological analysis of honey samples collected between 2006 and 2008. Both bees showed a polylectic foraging habit. Richness of pollen types per sample ranged between 10 and 34 (mean = 20.5 ± 7.7) for A. mellifera, and between 13 and 43 (mean = 24.8 ± 7.1) for T. fiebrigi. The 15 most abundant pollen types in the honeys of A. mellifera were, in decreasing order of importance, Euphorbiaceae, Euterpe edulis, Holocalyx balansae, Calyptranthes concinna-type, Actinostemon, Salix, Ruprechtia laxiflora, Myrcianthes pungens-type, Thinouia mucronata, Allophylus edulis, Ilex, Syagrus romanzoffiana, Gouania latifolia-type, Parapiptadenia rigida, and Baccharis-type, whereas in the honeys of T. fiebrigi the most important pollen types included S. romanzoffiana, Schinus weinmannifolius-type, Baccharis-type, H. balansae, E. edulis, Rhamnaceae, Citrus, Leonurus japonicus, G. latifolia-type, A. edulis, Gomphrena perennis-type, Pouteria gardneriana, P. rigida, Zanthoxylum, and Actinostemon.

Resumen Los recursos nectaríferos y poliníferos son clave para la supervivencia de las abejas sociales, por lo que identificar las fuentes de origen permite implementar acciones de manejo tendientes a un mayor suministro de nutrientes para las colonias. Asimismo, definir el origen floral de las mieles contribuye a la caracterización y comercialización de las mismas. El objetivo de nuestro trabajo fue identificar los recursos vegetales que proporcionan néctar a las colmenas de Apis mellifera y Tetragonisca fiebrigi en el norte de Misiones, a partir del análisis polínico de las muestras de miel colectadas entre 2006 y 2008. Ambas especies mostraron un hábito de alimentación poliléctico. La riqueza de tipos polínicos por muestra osciló entre 10 y 34 (media = 20,5 ± 7,7) para A. mellifera, y entre 13 y 43 (media = 24,8 ± 7,1) para T. fiebrigi. Los 15 tipos polínicos más abundantes en las mieles de A. mellifera fueron, en orden decreciente de importancia, Euphorbiaceae, Euterpe edulis, Holocalyx balansae, tipo Calyptranthes concinna, Actinostemon, Salix, Ruprechtia laxiflora, tipo Myrcianthes pungens, Thinouia mucronata, Allophylus edulis, Ilex, Syagrus romanzoffiana, tipo Gouania latifolia, Parapiptadenia rigida y tipo Baccharis, mientras que en las mieles de T. fiebrigi incluyeron a S. romanzoffiana, tipo Schinus weinmannifolius, tipo Baccharis, H. balansae, E. edulis, Rhamnaceae, Citrus, Leonurus japonicus, tipo G. latifolia, A. edulis, tipo Gomphrena perennis, Pouteria gardneriana, P. rigida, Zanthoxylum y Actinostemon.

Abstract In this study, we record for the first time the genus Notocyrtus (Heteroptera, Reduviidae) from Argentina based on three species: Notocyrtus dorsalis (Gray), Notocyrtus dispersus Carvalho & Costa, and Notocyrtus foveatus Stål. We also describe and illustrate a mimetic complex comprising the three Notocyrtus species and Tetragona clavipes (Fabricius) (Apidae, Meliponini), that were collected on Bahuinia forficata Link (Leguminosae: Caesalpinioideae). We include biological comments on the plant-reduvid-bee interaction and hypothesize about the functionality of the mimetic complex described.

Abstract Background Centruroides hirsutipalpus, of the family Buthidae, is a scorpion endemic to the Western Pacific region of Mexico. Although medically important, its venom has not yet been studied. Therefore, this communication aims to identify their venom components and possible functions. Methods Fingerprinting mass analysis of the soluble venom from this scorpion was achieved by high-performance liquid chromatography and electrospray mass spectrometry. Furthermore, the soluble venom and its toxic effects were evaluated extensively via electrophysiological assays in HEK cells expressing human voltage-gated Na+ channels (hNav 1.1 to Nav1.6), CHO cells expressing hNav 1.7, potassium channel hERG 1 (Ether-à-go-go-related-gene) and the human K+-channel hKv1.1. Results The separation of soluble venom produced 60 fractions from which 83 distinct components were identified. The molecular mass distribution of these components varies from 340 to 21,120 Da. Most of the peptides have a molecular weight between 7001 and 8000 Da (46% components), a range that usually corresponds to peptides known to affect Na+ channels. Peptides with molecular masses from 3000 to 5000 Da (28% of the components) were identified within the range corresponding to K+-channel blocking toxins. Two peptides were obtained in pure format and completely sequenced: one with 29 amino acids, showing sequence similarity to an "orphan peptide" of C. limpidus, and the other with 65 amino acid residues shown to be an arthropod toxin (lethal to crustaceans and toxic to crickets). The electrophysiological results of the whole soluble venom show a beta type modification of the currents of channels Nav1.1, Nav1.2 and Nav1.6. The main effect observed in channels hERG and hKv 1.1 was a reduction of the currents. Conclusion The venom contains more than 83 distinct components, among which are peptides that affect the function of human Na+-channels and K+-channels. Two new complete amino acid sequences were determined: one an arthropod toxin, the other a peptide of unknown function.

We analyzed contemporary oral narratives about birds collected among rural inhabitants of northern Córdoba in order to discuss alternative ways of interpreting human-bird relationships. A total of 150 stories were analyzed in terms of their typology, representativeness, ecological and social aspects expressed in them and functionality, as well as the utilitarian value of the species. In addition, we analyzed alternative ways of interpreting and classifying the augural role of birds. We propose that the narrative shows animistic traits (continuity between animals and humans) according to which the augural role of birds can be interpreted as a non-verbal communicative vestige between humans and avifauna. The mentioned birds (29 species) were grouped into 15 taxonomic orders and 22 families. The narratives classified as knowledge/practices made up 79.3% of the total of the non-narrative stories, while 19.5% corresponded to narrative stories. Ethological knowledges were among the best represented. Non-narrative stories referred to ecological knowledge about birds and practices related to their use as food. Myths, legends and anecdotes were the best represented narrative stories and, in general, they worked as transmitters of norms of social behaviours. The “ornithoauguries” can be explained as the interpretation of ecological knowledge of birds, as the elucidation of an intraspecific zoosemiotic message recoded by humans, or as an intersubjective communication between beings, as it happens in indigenous societies. Finally, we discuss the interpretation of these auguries in mestizo societies such as the creoles of northern Córdoba, which have indigenous and European influences.

Se analizaron narraciones orales contemporáneas sobre las aves recopiladas entre pobladores rurales del norte de Córdoba con el objetivo de debatir sobre las formas alternativas de interpretar las relaciones entre humanos y aves. Un total de 150 relatos fueron analizados en cuanto a su tipología, representatividad, aspectos ecológicos y sociales expresados en ellos y funcionalidad, así como el valor utilitario de las especies. Además, se analizaron formas alternativas de interpretar y clasificar el papel augural de las aves. Se propone que la narrativa evidencia rasgos animistas (continuidad entre animales y humanos) según los cuales el rol augural de las aves puede ser interpretado como un vestigio comunicativo no-verbal entre los humanos y la avifauna. Las aves mencionadas (29 especies) se agruparon en 15 órdenes y 22 familias taxonómicas. Las narraciones clasificadas como saberes/prácticas conformaron el 79.3% del total de los relatos no narrativos, mientras que el 19.5% correspondió a relatos narrativos. Entre los saberes mejor representados se destacan los etológicos. Los relatos no narrativos hicieron referencia a conocimientos ecológicos sobre las aves y a prácticas relacionadas a su utilización como alimento. Mitos, leyendas y anécdotas fueron los relatos narrativos mejor representados; en general, funcionaron como transmisores de normas de conductas sociales. Los “ornitoagurios” pueden explicarse como la interpretación de conocimientos ecológicos de las aves, como la elucidación de un mensaje zoosemiótico intraespecífico recodificado por los humanos o como una comunicación intersubjetiva entre seres, como ocurre en sociedades indígenas. Finalmente, se discute la interpretación de estos augurios en sociedades mestizas como la de los criollos del norte de Córdoba, que poseen influencias indígenas y europeas.

Abstract Background The cysteine-rich neurotoxins from elapid venoms are primarily responsible for human and animal envenomation; however, their low concentration in the venom may hamper the production of efficient elapid antivenoms. Therefore, the aim of the present study was to produce fully active elapid neurotoxic immunogens for elapid antivenom production. Method Cysteine-rich neurotoxins showed recombinant expression in two strains of E. coli, and were purified using affinity chromatography and reverse-phase HPLC (rpHPLC). Results The cDNA of the four disulfide-bridged peptide neurotoxin Mlat1 was cloned into a modified expression vector, pQE30, which was transfected into two different E. coli strains. The recombinant toxin (HisrMlat1) was found only in inclusion bodies in M15 strain cells, and in both inclusion bodies and cytoplasm in Origami strain cells. The HisrMlat1 from inclusion bodies from M15 cells was solubilized using guanidine hydrochloride, and then purified by rpHPLC. It showed various contiguous fractions having the same molecular mass, indicating that HisrMlat1 was oxidized after cell extraction forming different misfolded disulfide bridge arrangements without biological activity. In vitro folding conditions of the misfolded HisrMlat1 generated a biologically active HisrMlat1. On the other hand, the HisrMlat1 from the cytoplasm from Origami cells was already soluble, and then purified by HPLC. It showed a single fraction with neurotoxic activity; so, no folding steps were needed. The in vitro folded HisrMlat1 from M15 cells and the cytoplasmic soluble HisrMlat1from Origami cells were indistinguishable in their structure and neurotoxicity. Rabbit polyclonal antibodies raised up against biologically active HisrMlat1 recognized the native Mlat1 (nMlat1) from the whole venom of M. laticorallis. In addition, HisrMlat1 was recognized by horse polyclonal antibodies obtained from the immunization of elapid species from sub-Saharan Africa. Conclusion HisrMlat1 shows increased biological activities compared to the native peptide, and may be used as an immunizing agent in combination with other toxic components such phospholipases type A2 for elapid antivenom production.

Backgound The venom of the Cuban scorpion Rhopalurus junceus is poorly study from the point of view of their components at molecular level and the functions associated. The purpose of this article was to conduct a proteomic analysis of venom components from scorpions collected in different geographical areas of the country. Results Venom from the blue scorpion, as it is called, was collected separately from specimens of five distinct Cuban towns (Moa, La Poa, Limonar, El Chote and Farallones) of the Nipe-Sagua-Baracoa mountain massif and fractionated by high performance liquid chromatography (HPLC); the molecular masses of each fraction were ascertained by mass spectrometry analysis. At least 153 different molecular mass components were identified among the five samples analyzed. Molecular masses varied from 466 to 19755 Da. Scorpion HPLC profiles differed among these different geographical locations and the predominant molecular masses of their components. The most evident differences are in the relative concentration of the venom components. The most abundant components presented molecular weights around 4 kDa, known to be K+-channel specific peptides, and 7 kDa, known to be Na+-channel specific peptides, but with small molecular weight differences. Approximately 30 peptides found in venom samples from the different geographical areas are identical, supporting the idea that they all probably belong to the same species, with some interpopulational variations. Differences were also found in the presence of phospholipase, found in venoms from the Poa area (molecular weights on the order of 14 to 19 kDa). The only ubiquitous enzyme identified in the venoms from all five localities studied (hyaluronidase) presented the same 45 kD molecular mass, identified by gel electrophoresis analysis. Conclusions The venom of these scorpions from different geographical areas seem to be similar, and are rich in peptides that have of the same molecular masses of the peptides purified from other scorpions that affect ion-channel functions.