In this work, a methodology of synthesis was designed to obtain ZnO nanoparticles (ZnO NPs) in a controlled and reproducible manner. The nanoparticles obtained were characterized using infrared spectroscopy, X-ray diffraction, and transmission electron microscopy (TEM). Also, we determined the antifungal capacity in vitro of zinc oxide nanoparticles synthesized, examining their action on Erythricium salmonicolor fungy causal of pink disease. To determine the effect of the quantity of zinc precursor used during ZnO NPs synthesis on the antifungal capacity, 0.1 and 0.15 M concentrations of zinc acetate were examined. To study the inactivation of the mycelial growth of the fungus, different concentrations of ZnO NPs of the two types of synthesized samples were used. The inhibitory effect on the growth of the fungus was determined by measuring the growth area as a function of time. The morphological change was observed with high-resolution optical microscopy (HROM), while TEM was used to observe changes in its ultrastructure. The results showed that a concentration of 9 mmol L⁻¹ for the sample obtained from the 0.15 M and at 12 mmol L⁻¹ for the 0.1 M system significantly inhibited growth of E. salmonicolor. In the HROM images a deformation was observed in the growth pattern: notable thinning of the fibers of the hyphae and a clumping tendency. The TEM images showed a liquefaction of the cytoplasmic content, making it less electron-dense, with the presence of a number of vacuoles and significant detachment of the cell wall.

Colombia and Brazil are affected by severe cases of scorpionism. In Colombia the most dangerous accidents are caused by Tityus pachyurus that is widely distributed around this country. In the Brazilian Amazonian region scorpion stings are a common event caused by Tityus obscurus. The main objective of this work was to perform the molecular cloning of the putative Na⁺-channel scorpion toxins (NaScTxs) from T. pachyurus and T. obscurus venom glands and to analyze their phylogenetic relationship with other known NaScTxs from Tityus species. cDNA libraries from venom glands of these two species were constructed and five nucleotide sequences from T. pachyurus were identified as putative modulators of Na⁺-channels, and were named Tpa4, Tpa5, Tpa6, Tpa7 and Tpa8; the latter being the first anti-insect excitatory β-class NaScTx in Tityus scorpion venom to be described. Fifteen sequences from T. obscurus were identified as putative NaScTxs, among which three had been previously described, and the others were named To4 to To15. The peptides Tpa4, Tpa5, Tpa6, To6, To7, To9, To10 and To14 are closely related to the α-class NaScTxs, whereas Tpa7, Tpa8, To4, To8, To12 and To15 sequences are more related to the β-class NaScTxs. To5 is possibly an arthropod specific toxin. To11 and To13 share sequence similarities with both α and β NaScTxs. By means of phylogenetic analysis using the Maximum Parsimony method and the known NaScTxs from Tityus species, these toxins were clustered into 14 distinct groups. This communication describes new putative NaScTxs from T. pachyurus and T. obscurus and their phylogenetic analysis. The results indicate clear geographic separation between scorpions of Tityus genus inhabiting the Amazonian and Mountain Andes regions and those distributed over the Southern of the Amazonian rainforest. Based on the consensus sequences for the different clusters, a new nomenclature for the NaScTxs is proposed.

Background: Bothrops asperrepresents the clinically most important snake species in Central America and Northern South America, where it is responsible for an estimated 50–80% of snakebites. Compositional variability among the venom proteomes ofB.asperlineages across its wide range mirrors clinical differences in their envenomings. Bothropic antivenoms generated in a number of Latin American countries commonly exhibit a certain degree of paraspecific effectiveness in the neutralization of congeneric venoms. Defining the phylogeographic boundaries of an antivenom's effectivity has implications for optimizing its clinical use. However, the molecular bases and impact of venom compositions on the immune recognition and neutralization of the toxic activities of across geographically disparate populations ofB.asperlineages has not been comprehensively studied.Methodology/Principal findings: Third-generation antivenomics was applied to quantify the cross-immunorecognizing capacity against the individual components of venoms of threeB.asperlineages (B.asper(sensu stricto),B.ayerbeiandB.rhombeatus) distributed in south-western (SW) Colombia, of six Latin American antivenoms, produced against homologous (Colombia, INS-COL and PROBIOL) and Costa Rica (ICP)), and heterologous (Argentina (BIOL), Perú (INS-PERU) and Venezuela (UCV)) bothropic venoms.In vivoneutralization assays of the lethal, hemorrhagic, coagulant, defibrinogenating, myotoxic, edematogenic, indirect hemolytic, and proteolytic activities of the three SW ColombianB.asperlineage venoms were carried to compare the preclinical efficacy of three (Colombian INS-COL and PROBIOL, and Costa Rican ICP) antivenoms frequently used in Colombia. Antivenomics showed that all the six antivenom affinity matrices efficiently immunoretained most of theB.asperlineages venom proteins and exhibited impaired binding towards the venoms' peptidomes. The neutralization profile of the INS-COL, PROBIOL and ICP antivenoms towards the biological activities of the venoms of SW ColombianB.asper(sensu stricto),B.ayerbeiandB.rhombeatuslineages was coherent with the antivenomics outcome. In addition, the combination ofin vitro(antivenomics) andin vivoneutralization results allowed us to determine their toxin-specific and venom neutralizing antibody content. Noteworthy, heterologous INS-PERU, BIOL, and UCV bothropic antivenoms had equal or higher binding capacity towards the venoms components of SW ColombianB.asperlineages that the homologous Colombian and Costa Rican antivenoms.Conclusions/Significance: The combinedin vitroandin vivopreclinical outcome showed that antivenoms manufactured in Colombia and Costa Rica effectively neutralize the major toxic activities of SW ColombianB.asperlineage venoms. The antivenomics profiles of the heterologous antivenoms manufactured in Argentina, Venezuela, and Perú strongly suggests their (pre)clinical adequacy for the treatment ofB.asperlineage envenomings in SW Colombia. However, their recommendation in the clinical setting is pending onin vivoneutralization testing and clinical testing in humans.Bothrops asperis a highly adaptable snake species complex, which is considered the most dangerous snake throughout much of its distribution range from the Atlantic lowland of eastern México to northwestern Perú. Antivenoms are the only scientifically validated treatment of snakebite envenomings. Venom variation is particularly common in wide ranging species, such asB.asper, and may result in variable clinical presentations of envenomings, as is the case for theB.asperspecies complex, potentially undermining the efficacy of snakebite treatments depending on the immunization mixture used in the generation of the antivenom. Conversely, phylogenetic conservation of antigenic determinants confers an unpredictable degree of paraspecificity to homologous antivenoms produced for a geographic area, but also to heterologous congeneric antivenoms, towards the venom components of allopatric conspecific populations. This work aimed at comparing the preclinical profile of a panel of Latin American homologous and heterologous antivenoms against the venoms ofB.asperlineages distributed in SW Colombia. The outcome of this study strongly suggests the suitability of considering the heterologous antivenoms BIOL (Argentina), UCV (Venezuela) and INS-PERU (Perú) as alternatives to homologous Colombian INS-COL and PROBIOL and Costa Rican ICP antivenoms for the treatment of envenomings byB.asper(sensu stricto) in W Colombia and Ecuador,B.ayerbeiin Cauca and Nariño (Colombia), andB.rhombeatusin Cauca river valley, SW Colombia.