Analysis of High Molecular Mass Compounds from the Spider Pamphobeteus verdolaga Venom Gland. A Transcriptomic and MS ID Approach.

Nowadays, spider venom research focuses on the neurotoxic activity of small peptides. In this study, we investigated high-molecular-mass compounds that have either enzymatic activity or housekeeping functions present in either the venom gland or venom of Pamphobeteus verdolaga. We used proteomic and transcriptomic-assisted approaches to recognize the proteins sequences related to high-molecular-mass compounds present in either venom gland or venom. We report the amino acid sequences (partial or complete) of 45 high-molecular-mass compounds detected by transcriptomics showing similarity to other proteins with either enzymatic activity (i.e., phospholipases A2, kunitz-type, hyaluronidases, and sphingomyelinase D) or housekeeping functions involved in the signaling process, glucanotransferase function, and beta-N-acetylglucosaminidase activity. MS/MS analysis showed fragments exhibiting a resemblance similarity with different sequences detected by transcriptomics corresponding to sphingomyelinase D, hyaluronidase, lycotoxins, cysteine-rich secretory proteins, and kunitz-type serine protease inhibitors, among others. Additionally, we report a probably new protein sequence corresponding to the lycotoxin family detected by transcriptomics. The phylogeny analysis suggested that P. verdolaga includes a basal protein that underwent a duplication event that gave origin to the lycotoxin proteins reported for Lycosa sp. This approach allows proposing an evolutionary relationship of high-molecular-mass proteins among P. verdolaga and other spider species.

MS/MS analysis of four scorpion venoms from Colombia: a descriptive approach.

BACKGROUND: Scorpions are widely known for the neurotoxic effects of their venoms, which contain peptides affecting ionic channels. Although Colombia is recognized for its scorpion diversity, only a few studies are available describing the venom content. METHODS: In this descriptive study, we analyzed the MS/MS sequence, electrophoretic and chromatographic profile linked to a bioinformatics analysis of the scorpions Chactas reticulatus (Chactidae), Opisthacanthus elatus (Hormuridae), Centruroides edwardsii (Buthidae) and Tityus asthenes (Buthidae) from Colombia. RESULTS: Each scorpion showed a specific electrophoretic and chromatographic profile. The electrophoretic profiles indicate the presence of high molecular mass compounds in all venoms, with a predominance of low molecular mass compounds in the Buthidae species. Chromatographic profiles showed a similar pattern as the electrophoretic profiles. From the MS/MS analysis of the chromatographic collected fractions, we obtained internal peptide sequences corresponding to proteins reported in scorpions from the respective family of the analyzed samples. Some of these proteins correspond to neurotoxins affecting ionic channels, antimicrobial peptides and metalloproteinase-like fragments. In the venom of Tityus asthenes, the MSn analysis allowed the detection of two toxins affecting sodium channels covering 50% and 84% of the sequence respectively, showing 100% sequence similarity. Two sequences from Tityus asthenes showed sequence similarity with a phospholipase from Opisthacanthus cayaporum indicating the presence of this type of toxin in this species for the first time. One sequence matching a hypothetical secreted protein from Hottentotta judaicus was found in three of the studied venoms. We found that this protein is common in the Buthidae family whereas it has been reported in other families - such as Scorpionidae - and may be part of the evolutionary puzzle of venoms in these arachnids. CONCLUSION: Buthidae venoms from Colombia can be considered an important source of peptides similar to toxins affecting ionic channels. An interesting predicted antimicrobial peptide was detected in three of the analyzed venoms.

MS/MS analysis of four scorpion venoms from Colombia: a descriptive approach

Background: Scorpions are widely known for the neurotoxic effects of their venoms, which contain peptides affecting ionic channels. Although Colombia is recognized for its scorpion diversity, only a few studies are available describing the venom content. Methods: In this descriptive study, we analyzed the MS/MS sequence, electrophoretic and chromatographic profile linked to a bioinformatics analysis of the scorpions Chactas reticulatus (Chactidae), Opisthacanthus elatus (Hormuridae), Centruroides edwardsii (Buthidae) and Tityus asthenes (Buthidae) from Colombia. Results: Each scorpion showed a specific electrophoretic and chromatographic profile. The electrophoretic profiles indicate the presence of high molecular mass compounds in all venoms, with a predominance of low molecular mass compounds in the Buthidae species. Chromatographic profiles showed a similar pattern as the electrophoretic profiles. From the MS/MS analysis of the chromatographic collected fractions, we obtained internal peptide sequences corresponding to proteins reported in scorpions from the respective family of the analyzed samples. Some of these proteins correspond to neurotoxins affecting ionic channels, antimicrobial peptides and metalloproteinase-like fragments. In the venom of Tityus asthenes, the MSn analysis allowed the detection of two toxins affecting sodium channels covering 50% and 84% of the sequence respectively, showing 100% sequence similarity. Two sequences from Tityus asthenes showed sequence similarity with a phospholipase from Opisthacanthus cayaporum indicating the presence of this type of toxin in this species for the first time. One sequence matching a hypothetical secreted protein from Hottentotta judaicus was found in three of the studied venoms. We found that this protein is common in the Buthidae family whereas it has been reported in other families - such as Scorpionidae - and may be part of the evolutionary puzzle of venoms in these arachnids. Conclusion: Buthidae venoms from Colombia can be considered an important source of peptides similar to toxins affecting ionic channels. An interesting predicted antimicrobial peptide was detected in three of the analyzed venoms.(AU)

Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels.

Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium influx assays, tandem mass spectrometry analysis (tMS/MS), and venom-gland transcriptome. rp-HPLC fractions of P. verdolaga venom showed activity on CaV2.2, CaV3.2, and NaV1.7 ion channels. Active fractions contained several peptides with molecular masses ranging from 3399.4 to 3839.6 Da. The tMS/MS analysis of active fraction displaying the strongest activity to inhibit calcium channels showed sequence fragments similar to one of the translated transcripts detected in the venom-gland transcriptome. The putative peptide of this translated transcript corresponded to a toxin, here named ω-theraphositoxin-Pv3a, a potential ion channel modulator toxin that is, in addition, very similar to other theraphositoxins affecting calcium channels (i.e., ω-theraphotoxin-Asp1a). Additionally, using this holistic approach, we found that P. verdolaga venom is an important source of disulfide-rich proteins expressing at least eight superfamilies.

DISTRIBUCIÓN DE Centruroides edwardsii (GERVAIS, 1843) EN EL DEPARTAMENTO EN ANTIOQUIA, COLOMBIA / Distribution of Centruroides edwardsii (GERVAIS, 1843) in the Antioquia Province, Colombia

Se reporta la distribución de Centruroides edwardsii (Gervais, 1843) para el departamento de Antioquia, y el área urbana de su capital político-administrativa, la ciudad de Medellín, Colombia, con base a material depositado en la colección del Serpentario de la Universidad de Antioquia (SUA). Se estableció dimorfismo sexual en C. edwardsii, mediante análisis de escalamiento multidimensional y el modelo ANOSIM, encontrando que los machos son más grandes que las hembras. Se reporta para el área zona centro y nor-oriental de Medellín una importante colonia de este escorpión, hecho de gran relevancia debido a la importancia clínica de este escorpión. Se amplía además el rango de distribución desde el nivel del mar hasta los 2200 m s.n.m.

We report the distribution of the Centruroides edwardsii (Gervais, 1843) in the Antioquia province, and the urban area of the capital, Medellín, Colombia, based in the material examined in the collection of the Universidad de Antioquia Serpentarium (SUA). A sexual dimporphism were established in C. edwardsii when a multidimensional analysis and using for the statistical significance the ANOSIM model were performed founding higher sizes in males. An important colony of this scorpion is reported in the central and nor-oriental zone of Medellín, aspect that is very relevant since this scorpion is considered clinically relevante. The species distribution rank is also expanded from the sea level, up to 2200 m a.s.l.

Phenotypic differences in a cryptic predator: factors influencing morphological variation in the terciopelo Bothrops asper (Garman, 1884; Serpentes: Viperidae).

The terciopelo Bothrops asper, is a cryptic lancehead pitviper widely distributed in humid environments of Middle America and the north-western portion of South America. Throughout its extensive distribution range, the terciopelo exhibits great morphological variation in external characters, a situation that has complicated its proper separation from other related species. In this paper, we analyzed the phenotypic variation of B. asper based in a sample of 514 specimens from nine distinct physiographic regions. Univariate and multivariate analyses demonstrated great phenotypic differentiation among most pre-established groups, and the pattern is fairly congruent between sexes. However, no correspondence was observed between morphological variation and molecular divergence, inferred from mDNA sequences, between individuals representing the physiographical regions under study. Geographic variation in the number of interrictals, ventral scales, subcaudal scales and dorsal blotches was positively correlated with latitude and number of dry months, but inversely related with precipitation. However, other variables do not exhibit such an effect. The observed relationships between scale counts and environmental variables are explained in terms of selective pressures to improve water balance along the distributional range of the species.

Aspectos toxinológicos, clínicos y epidemiológicos el envenenamiento producido por el escorpión Tityus fuhrmanni Kraepelin / Toxinological, clinical, and epidemiological aspects of poisoning produced by the Titys fuhrmanni Kraepelin scorpion

Con el objetivo de determinar las características toxinológicas, clínicas y epidemiológicas del envenenamiento producido por el escorpión T. fuhrmanni, se realizó durante un año un estudio prospectivo en un sector de la ciudad de Medellín (cerro El Volador y los barrios aledaños San Germán y El Volador), en una muestra del 9,6 por ciento de las casas (180) y del 9,4 por ciento de la población del área (719 habitantes). Se incluyó también la recolección de especímenes vivos (128) y de veneno para la parte experimental. La dispersión de escorpiones para el área de estudio fue del 100 por ciento, con alta infestación domiciliaria en las viviendas del cerro y del barrio San Germán (100 por ciento y 32,3 porciento, respectivamente). Durante todo el estudio se detectaron 32 accidentes por T. fuhrmanni, 7 (21,9 por ciento) en niños, 15 en el cerro y 10 en el barrio San Germán, con tasas de ataque del 83 por ciento y del 3,9 porciento, respectivamente. Dieciocho (56,3 por ciento) accidentes ocurrieron en el interior de las viviendas, principalmente en las manos (31,3 por ciento), la cabeza y el cuello (18,8 por ciento) y durante la noche o en las primeras horas de la mañana (62,5 por ciento). El 90,6 por ciento de los casos fueron leves y el 9,4 por ciento (3 niños) tuvieron envenenamiento moderado con signos sistémicos (sudoración generalizada, dolor abdominal). No hubo casos graves y los pacientes no requirieron hospitalización. Tityus fuhrmanni produjo 0,56+/-0,27 mg de veneno por ordeño, y su DL50 (i.p.) en ratones (18-20 g) fue 79,2 ug (3,9 mg veneno/kg). Los animales presentaron sialorrea, piloerección, somnolencia, sudoración generalizada, taquipnea, cianosis, ataxia y convulsiones antes de morir. La rápida aparición (10-15 min) de los signos de envenenamiento en los ratones y su pronta desaparición (2 horas) en los sobrevivientes, permiten concluir que el veneno es de rápida absorción, distribución y eliminación

Los escorpiones: aspectos ecológicos, biológicos y toxinológicos / Scorpions: ecological, biological and toxinological aspects

Los escorpiones son artrópodos quelicerados distribuidos en las regiones tropicales y subtropicales del mundo. Su veneno es una mezcla bioquímica compleja, formada principalmente por proteínas neurotóxicas de bajo peso molecular que ejercen su acción sobre los canales iónicos, ocasionando efectos nocivos en sus víctimas. En algunos países como Argelia, Túnez, Irán, Méjico y Brasil, el escorpionismo es un problema de salud pública. Las investigaciones encaminadas a contrarrestar tales efectos se han dirigido a la purificación de estas toxinas, a determinar las secuencia nucleotídicas de los genes que las codifican y a entender los mecanismos asociados con la regulación y expresión de los genes. A su vez, a establecer si hay reacción cruzada entre los venenos de diferentes especies de escorpiones, a determinar la acción farmacológica del veneno, las manifestaciones clínicas del envenenamiento y el uso de antivenenos como protocolo de tratamiento. Estas toxinas tambien han sido herramientas invaluables en la identificación, purificación, mapeo estructural y caracterización funcional de los canales iónicos. Este artículo de revisión presenta varios aspectos sobre el conocimiento actual de los escorpiones, tanto en sus caracteres ecológicos y biológicos, como e la composición y mecanismos de acción de su veneno