Bothrops Neuwiedi Research Articles

Designing 1,2,3‐Triazole Derivatives for Targeted Inhibition of Proteolytic and Coagulant Activities in Bothrops Jararaca and Bothrops neuwiedi Snake Venoms

AbstractSnakebites inflict significant injuries on humans and animals, posing a severe global public health challenge due to high mortality and morbidity rates. This study presents results against the venoms of Bothrops jararaca and Bothrops neuwiedi using sixteen synthetic hybrid molecules containing thiophene and triazoles, designated as 6a–h and 7a–h. The compounds were assessed for their ability to inhibit proteolytic and plasma coagulant activities induced by the snake venoms, and their toxicity was analyzed. Most derivatives demonstrated nontoxicity through in silico analysis with the Osiris Property Explorer tool and in vitro cytotoxic hemocompatibility on red blood cells. Notably, only derivative 6e exhibited toxicity to erythrocytes, while 6b, 6d, 7c, and 7f displayed mutagenic, tumorigenic, or adverse effects on the reproductive system. Derivatives 6a, 6c–h, 7c–e, 7g, or 7h effectively inhibited the coagulating activity of B. jararaca venom, while 6a–b, 6d–e, 6h, or 7a–f inhibited coagulation induced by B. neuwiedi venom. All derivatives demonstrated inhibition of the proteolytic activity caused by B. jararaca venom, and derivatives 6a–d and 7e–f inhibited the activity of B. neuwiedi venom. Consequently, these derivatives exhibited varying degrees of efficacy in countering two crucial toxic effects of B. jararaca or B. neuwiedi venoms, suggesting their potential as antivenoms against both species.

Arsenic and metal levels in snake tissues from Lagoa Santa Karst, Brazil.

Concentrations of one metalloid (As) and eight metals (Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn) were determined in tissues (muscle, liver, and kidney) of eight snake species (Bothrops neuwiedi, Crotalus durissus, Dipsas mikanii, Epicrates crassus, Helicops modestus, Micrurus carvalhoi, Oxyrhopus guibei, and Oxyrhopus trigeminus) from Lagoa Santa Karst. Except for Cu and Zn, all other analyzed elements were detected in concentrations within the ranges previously reported for snakes inhabiting polluted areas, emphasizing Hg (specific Hg mean concentrations varied from 0.87 to 9.76μgg-1 d.w). The highest mean concentrations of all elements except Zn were found in muscle samples of the false corals O. guibei (means ranged from 2.01 [Pb] to 9.76 [Hg]). The highest Zn mean concentration (13.77μgg-1 d.w) was detected in the kidney of the water snake H. modestus. No significant correlation was found between element concentrations and body size for all species. Significant interorgan differences were observed for As, Cr, Cu, Hg, Mn, Pb, and Zn concentrations in the three tissues in H. modestus. Significant interspecific differences were found in at least one organ for all elements. Significant pairwise differences were found between diet specialist species and between these species and broader diet species, while no significant difference was found between the broader diet species. The bioaccumulation of As and metals in snakes from Lagoa Santa Karst could be associated with natural rock dissolution and erosion processes but also with the wide-scale mining in the region and the increased agriculture and urbanization.

Profilings of subproteomes of lectin-binding proteins of nine Bothrops venoms reveal variability driven by different glycan types

Snake venom proteomes have long been investigated to explore a multitude of biologically active components that are used for prey capture and defense, and are involved in the pathological effects observed upon mammalian envenomation. Glycosylation is a major protein post-translational modification in venoms and contributes to the diversification of proteomes. We have shown that Bothrops venoms are markedly defined by their content of glycoproteins, and that most N-glycan structures of eight Bothrops venoms contain sialic acid, while bisected N-acetylglucosamine was identified in Bothrops cotiara venom. To further investigate the mechanisms involved in the generation of different venoms by related snakes, here the glycoproteomes of nine Bothrops venoms (Bothrops atrox, B. cotiara, Bothrops erythromelas, Bothrops fonsecai, B. insularis, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi) were comparatively analyzed by enrichment with three lectins of different specificities, recognizing bisecting N-acetylglucosamine- and sialic acid-containing glycoproteins, and mass spectrometry. The lectin capture strategy generated venom fractions enriched with several glycoproteins, including metalloprotease, serine protease, and L- amino acid oxidase, in addition to various types of low abundant enzymes. The different contents of lectin-enriched proteins underscore novel aspects of the variability of the glycoprotein subproteomes of Bothrops venoms and point to the role of distinct types of glycan chains in generating different venoms by closely related snake species.

Differential coagulotoxicity of metalloprotease isoforms from Bothrops neuwiedi snake venom and consequent variations in antivenom efficacy

Bothrops (lance-head pit vipers) venoms are rich in weaponised metalloprotease enzymes (SVMP). These toxic enzymes are structurally diverse and functionally versatile. Potent coagulotoxicity is particularly important for prey capture (via stroke-induction) and relevant to human clinical cases (due to consumption of clotting factors including the critical depletion of fibrinogen). In this study, three distinct isoforms of P-III class SVMPs (IC, IIB and IIC), isolated from Bothrops neuwiedi venom, were evaluated for their differential capacities to affect hemostasis of prey and human plasma. Furthermore, we tested the relative antivenom neutralisation of effects upon human plasma. The toxic enzymes displayed differential procoagulant potency between plasma types, and clinically relevant antivenom efficacy variations were observed. Of particular importance was the confirmation the antivenom performed better against prothrombin activating toxins than Factor X activating toxins, which is likely due to the greater prevalence of the former in the immunising venoms used for antivenom production. This is clinically relevant as the enzymes displayed differential potency in this regard, with one (IC) in particular being extremely potent in activating Factor X and thus was correspondingly poorly neutralised. This study broadens the current understanding about the adaptive role of the SVMPs, as well as highlights how the functional diversity of SVMP isoforms can influence clinical outcomes.Key Contribution: Our findings shed light upon the hemorrhagic and coagulotoxic effects of three SVMPs of the P-III class, as well as the coagulotoxic effects of SVMPs on human, avian and amphibian plasmas. Antivenom neutralised prothrombin-activating isoforms better than Factor X activating isoforms.

Bothrops neuwiedi snake venom: Ontogenetic characterization and imunorecognition by bothropic antivenom

Bothrops neuwiedi snake venom: Ontogenetic characterization and imunorecognition by bothropic antivenom

B. neuwiedi complex: What the venom of these snake species have in common? Compositional and functional investigation of venoms from Bothrops neuwiedi complex

B. neuwiedi complex: What the venom of these snake species have in common? Compositional and functional investigation of venoms from Bothrops neuwiedi complex

A complex biogeographic history of diversification in Neotropical lancehead pitvipers (Serpentes, Viperidae)

AbstractBased on the literature, we had predicted that the diversification within the Neotropical snake genusBothropsoccurred along a latitudinal gradient from north to south, with diversification into unoccupied niches through ecological opportunity, not correlated with geoclimatic events. Using a dated phylogeny and estimating likelihoods of ancestral states at cladogenesis events, we reconstructed ancestral areas and assessed major events of the diversification ofBothropsclades, and we also discuss systematic implications for this group. Based on the phylogeny we produced,B. lojanuswas not considered as part of the genusBothropssince the results recovered this species nested within theBothrocophiasclade. We infer that the diversification of the MioceneBothrops pictusandBothrops alternatusclades may be related to the uplift of the western slopes of the Andes and the Argentinian Patagonian Andes, respectively. The PlioceneBothrops taeniatusandBothrops osborneiclades may be related to the uplift of the eastern and northern Andes, respectively. The Plio‐PleistoceneBothrops neuwiediclade may be related to the habitat transitions from a warmer and forested environment to a cooler and open landscape; theBothrops jararaca(i.e. island endemic species) andBothrops lanceolatusclades to over‐water dispersal with island speciation; andBothrops atroxclade to the appearance of the Panamanian land bridge. We found that a multitemporal and multidirectional history of diversification may be correlated with geoclimatic and dispersalist events. We argue that the vacant niche hypothesis by itself does not explainBothropsdiversification.

Leukocyte recruitment induced by snake venom metalloproteinases: Role of the catalytic domain

Snake venom metalloproteinases (SVMPs) are key toxins involved in local inflammatory reactions after snakebites. This study aimed to investigate the effect of SVMP domains on the alterations in leukocyte-endothelium interactions in the microcirculation of mouse cremaster muscle. We studied three toxins: BnP1, a PI-toxin isolated from Bothrops neuwiedi venom, which only bears a catalytic domain; Jararhagin (Jar), a PIII-toxin isolated from Bothrops jararaca venom with a catalytic domain, as well as ECD-disintegrin and cysteine-rich domains; and Jar-C, which is produced from the autolysis of Jar and devoid of a catalytic domain. All these toxins induced an increase in the adhesion and migration of leukocytes. By inhibiting the catalytic activity of Jar and BnP1 with 1.10-phenanthroline (oPhe), leukocytes were no longer recruited. Circular dichroism analysis showed structural changes in oPhe-treated Jar, but these changes were not enough to prevent the binding of Jar to collagen, which occurred through the ECD-disintegrin domain. The results showed that the catalytic domain of SVMPs is the principal domain responsible for the induction of leukocyte recruitment and suggest that the other domains could also present inflammatory potential only when devoid of the catalytic domain, as with Jar-C.

A new species of Bothrops (Serpentes: Viperidae: Crotalinae) from Pampas del Heath, southeastern Peru, with comments on the systematics of the Bothrops neuwiedi species group.

We describe a new species of pitviper of the genus Bothrops from the Peruvian Pampas del Heath, in the Bahuaja-Sonene National Park. Pampas del Heath is an area of seasonally flooded savannas and a northwestern extension of the Gran Chaco Boliviano-Paraguayo. The new species is easily distinguished from its congeners by the exclusive combination of dorsal color pattern of body consisting of small C-shaped blotches, postocular stripe originating posteriorly to the eye, covering posterior supralabials, dorsum of the head with paired markings arranged symmetrically, venter cream heavily speckled with brown, prelacunal scale discrete in contact with second supralabial, three to five prefoveals, subfoveal single usually present, postfoveals absent to two, canthals two, seven intersupraoculars, one or two suboculars, two or three postoculars, seven or eight supralabials, nine to eleven infralabials, 26-27 interrictals, 23-25 middorsal scales, 172 ventrals in the female and 169-173 in males, 45 subcaudals in the female and 50 in males. We performed separate and combined phylogenetic analyses based on morphology and five mitochondrial genes and recovered the new species as a member of the Bothrops neuwiedi species group. All lineages of this clade inhabit the South American dry diagonal. This novel species of pitviper increases the known diversity of the genus Bothrops and adds to the number of described taxa from the unique and scarcely known ecosystem of Pampas del Heath.

Comparative study of protein profile and enzymatic activities of venoms from snake species that compose the Bothrops neuwiedi complex

Comparative study of protein profile and enzymatic activities of venoms from snake species that compose the Bothrops neuwiedi complex

Communities and occurrences of Squamata reptiles in different vegetation types of the Serra de São José, Minas Gerais, Brazil

Abstract The objective of the present study was to learn which species of Squamata reptiles occur in Protected Area São José, in Tiradentes, Minas Gerais, Brazil. Between November 2009 and December 2010 reptiles were captured. In total 157 specimens were recorded of 29 species, 16 snakes, 12 lizards and one amphisbaena. Among the snakes, Dipsadidae showed the greatest richness, with a total of nine species. The group of snakes had the highest number of species present in the community, but 79% of sampled specimens were lizards, Enyalius bilineatus being the most abundant species, with 21% of occurrence. The area with the highest richness was the Cerradão. The lower abundance was found in the Gallery Forest area (n=14), but it was the vegetation type with the highest equitability. Areas of Cerradão and Cerrado sensu strictu showed the most similarity. In these areas five species were recorded in common, Bothrops neuwiedi (n=3) being the only species of snake, and the two species of lizards most abundant in both environments were Enyalius bilineatus (n=32) and Ameivula ocellifera (n=19). Ophiodes striatus and Xenodon merremii were common to Cerradão and Dirty Field areas. There was no species recorded that were common to the environments of Cerrado and Dirty Field but two species not sympatric were found of the same genus, Tropidurus torquatus, which was found only in the Cerrado sensu strictu and Tropidurus itambere exclusively in Dirty Field. Since none of the rarefaction curves reached full asymptote, this highlights the need for further study due to the high probability of new species being recorded for the studied area.

Ontogenetic Variation in Biological Activities of Venoms from Hybrids between Bothrops erythromelas and Bothrops neuwiedi Snakes.

Lance-headed snakes are found in Central and South America, and they account for most snakebites in Brazil. The phylogeny of South American pitvipers has been reviewed, and the presence of natural and non-natural hybrids between different species of Bothrops snakes demonstrates that reproductive isolation of several species is still incomplete. The present study aimed to analyze the biological features, particularly the thrombin-like activity, of venoms from hybrids born in captivity, from the mating of a female Bothrops erythromelas and a male Bothrops neuwiedi, two species whose venoms are known to display ontogenetic variation. Proteolytic activity on azocoll and amidolytic activity on N-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BAPNA) were lowest when hybrids were 3 months old, and increased over body growth, reaching values similar to those of the father when hybrids were 12 months old. The clotting activity on plasma diminished as hybrids grew; venoms from 3- and 6-months old hybrids showed low clotting activity on fibrinogen (i.e., thrombin-like activity), like the mother venom, and such activity was detected only when hybrids were older than 1 year of age. Altogether, these results point out that venom features in hybrid snakes are genetically controlled during the ontogenetic development. Despite the presence of the thrombin-like enzyme gene(s) in hybrid snakes, they are silenced during the first six months of life.

Partial in vitro analysis of toxic and antigenic activities of eleven Peruvian pitviper snake venoms

This work used eleven Peruvian snake venoms (Bothrops andianus, Bothrops atrox, Bothrops barnetti, Bothrops castelnaudi, Bothriopsis chloromelas, Bothrocophias microphthalmus, Bothrops neuwiedi, Bothriopsis oligolepis, Bothriopsis peruviana, Bothrops pictus and Bothriopsis taeniata) to perform in vitro experimentation and determine its main characteristics. Hyaluronidase (HYAL), phospholipase A2 (PLA2), snake venom metalloproteinase (SVMP), snake venom serine protease (SVSP) and l-amino acid oxidase (LAAO) activities; toxicity by cell viability assays using MGSO3, VERO and HeLa cell lineages; and crossed immunoreactivity with Peruvian (PAV) and Brazilian (BAV) antibothropic polyvalent antivenoms, through ELISA and Western Blotting assays, were determined. Results show that the activities tested in this study were not similar amongst the venoms and each species present their own peculiarities, highlighting the diversity within Bothrops complex. All venoms were capable of reducing cell viability of all tested lineages. It was also demonstrated the crossed recognition of all tested venoms by both antivenoms.

Tabebuia aurea decreases inflammatory, myotoxic and hemorrhagic activities induced by the venom of Bothrops neuwiedi

Ethnopharmacological relevanceEthnobotanical studies show that Tabebuia aurea has been used as anti-inflammatory and for snake bite. Evaluate the effect of treatment with the hydroethanolic extract of Tabebuia aurea (HETa) on inflammatory, hemorrhagic and myotoxic activities induced by Bothrops neuwiedi (BnV) in mice. Materials and methodsThe anti-inflammatory, antihemorragic and antimyotoxic properties of the HETa 100, 200 and 400mg/kg or BnV neutralized with HETa (1:50) were evaluated using the following animal models: BnV-induced paw edema, BnV-induced recruitment of polymorphonuclear cells into the peritoneal cavity, hemorrhagic activity, myotoxic activity and hydrogen peroxide production by peritoneal macrophages in vitro. ResultsHETa inhibited the paw edema and polymorphonuclear cell recruitment into the peritoneal cavity. BnV neutralized with HETa reduced the hemorrhagic activity and histopathological analysis of skeletal muscle tissue showed that the hemorrhagic area was smaller and multifocal. The leukocyte infiltrate was less intense and muscle necrosis discrete. BnV induced hydrogen peroxide production and BnV neutralized reduced this production. In addition, the HETa was nontoxic to macrophages. ConclusionsThe activities of the HETa presented herein justify the popular use of Tabebuia aurea in inflammatory situations from snake bite.

Phylogenetic relationships within Bothrops neuwiedi group (Serpentes, Squamata): Geographically highly-structured lineages, evidence of introgressive hybridization and Neogene/Quaternary diversification

Eight current species of snakes of the Bothrops neuwiedi group are widespread in South American open biomes from northeastern Brazil to southeastern Argentina. In this paper, 140 samples from 93 different localities were used to investigate species boundaries and to provide a hypothesis of phylogenetic relationships among the members of this group based on 1122bp of cyt b and ND4 from mitochondrial DNA and also investigate the patterns and processes occurring in the evolutionary history of the group. Combined data recovered the B. neuwiedi group as a highly supported monophyletic group in maximum parsimony, maximum likelihood and Bayesian analyses, as well as four major clades (Northeast I, Northeast II, East–West, West-South) highly-structured geographically. Monophyly was recovered only for B. pubescens. By contrast, B. diporus, B. lutzi, B. erythromelas, B. mattogrossensis, B. neuwiedi, B. marmoratus, and B. pauloensis, as currently defined on the basis of morphology, were polyphyletic. Sympatry, phenotypic intergrades and shared mtDNA haplotypes, mainly between B. marmoratus and B. pauloensis suggest recent introgressive hybridization and the possible occurrence of a narrow hybrid zone in Central Brazil. Our data suggest at least three candidate species: B. neuwiedi from Espinhaço Range, B. mattogrossensis (TM173) from Serra da Borda (MT) and B. diporus (PT3404) from Castro Barros, Argentina. Divergence estimates highlight the importance of Neogene events in the origin of B. neuwiedi group, and the origin of species and diversification of populations of the Neotropical fauna from open biomes during the Quaternary climate fluctuations. Data reported here represent a remarkable increase of the B. neuwiedi group sampling size, since representatives of all the current recognized species from a wide geographic range are included in this study, providing basic information for understanding the evolution and conservation of Neotropical biodiversity.

In vitro comparison of enzymatic effects among Brazilian Bothrops spp. venoms

In various types of snake venom, the major toxic components are proteinases and members of the phospholipase A2 family, although other enzymes also contribute to the toxicity. In this study, we evaluated the proteolytic, phospholipase, and l-Amino acid oxidase activities in the venom of five Bothrops species—Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi, and Bothrops alternatus—all of which are used in the production of commercial antivenom, prepared in horses. The enzymatic activities of each species' venom were classified as high, moderate, or low. B. moojeni venom demonstrated the highest enzymatic activity profile, followed by the venom of B. neuwiedi, B. jararacussu, B. jararaca, and B. alternatus. To our knowledge, this is the first study to compare all of these enzymes from multiple species, which is significant in view of the activity of l-amino acid oxidase across Bothrops species.

Umbelliferone induces changes in the structure and pharmacological activities of Bn IV, a phospholipase A 2 isoform isolated from Bothrops neuwiedi

In this paper was demonstrated that umbelliferone induces changes in structure and pharmacological activities of Bn IV, a lysine 49 secretory phospholipase A 2 (sPLA2) from Bothrops neuwiedi. Incubation of Bn IV with umbelliferone virtually abolished platelet aggregation, edema, and myotoxicity induced by native Bn IV. The amino acid sequence of Bn IV showed high sequence similarities with other Lys49 sPLA2s from B. jararacussu (BthTx-I), B. pirajai (PrTx-I), and B. neuwiedi pauloensis (Bn SP6 and Bn SP7). This sPLA2 also has a highly conserved C-terminal amino acid sequence, which has been shown as important for the pharmacological activities of Lys49 sPLA2. Sequencing of Bn IV previously treated with umbelliferone revealed modification of S(1) and S(20). Fluorescent spectral analysis and circular dichroism (CD) studies showed that umbelliferone modified the secondary structure of this protein. Moreover, the pharmacological activity of Bn IV is driven by synergism of the C-terminal region with the α-helix motifs, which are involved in substrate binding of the Asp49 and Lys49 residues of sPLA2 and have a direct effect on the Ca 2+-independent membrane damage of some secretory snake venom PLA2. For Bn IV, these interactions are potentially important for triggering the pharmacological activity of this sPLA2.

Diversity of metalloproteinases in Bothrops neuwiedi snake venom transcripts: evidences for recombination between different classes of SVMPs

BackgroundSnake venom metalloproteinases (SVMPs) are widely distributed in snake venoms and are versatile toxins, targeting many important elements involved in hemostasis, such as basement membrane proteins, clotting proteins, platelets, endothelial and inflammatory cells. The functional diversity of SVMPs is in part due to the structural organization of different combinations of catalytic, disintegrin, disintegrin-like and cysteine-rich domains, which categorizes SVMPs in 3 classes of precursor molecules (PI, PII and PIII) further divided in 11 subclasses, 6 of them belonging to PII group. This heterogeneity is currently correlated to genetic accelerated evolution and post-translational modifications.ResultsThirty-one SVMP cDNAs were full length cloned from a single specimen of Bothrops neuwiedi snake, sequenced and grouped in eleven distinct sequences and further analyzed by cladistic analysis. Class P-I and class P-III sequences presented the expected tree topology for fibrinolytic and hemorrhagic SVMPs, respectively. In opposition, three distinct segregations were observed for class P-II sequences. P-IIb showed the typical segregation of class P-II SVMPs. However, P-IIa grouped with class P-I cDNAs presenting a 100% identity in the 365 bp at their 5' ends, suggesting post-transcription events for interclass recombination. In addition, catalytic domain of P-IIx sequences segregated with non-hemorrhagic class P-III SVMPs while their disintegrin domain grouped with other class P-II disintegrin domains suggesting independent evolution of catalytic and disintegrin domains. Complementary regions within cDNA sequences were noted and may participate in recombination either at DNA or RNA levels. Proteins predicted by these cDNAs show the main features of the correspondent classes of SVMP, but P-IIb and P-IIx included two additional cysteines cysteines at the C-termini of the disintegrin domains in positions not yet described.ConclusionsIn B. neuwiedi venom gland, class P-II SVMPs were represented by three different types of transcripts that may have arisen by interclass recombination with P-I and P-III sequences after the divergence of the different classes of SVMPs. Our observations indicate that exon shuffling or post-transcriptional mechanisms may be driving these recombinations generating new functional possibilities for this complex group of snake toxins.

Crystal structure of Bn IV in complex with myristic acid: A Lys49 myotoxic phospholipase A2 from Bothrops neuwiedi venom

The LYS49-PLA2s myotoxins have attracted attention as models for the induction of myonecrosis by a catalytically independent mechanism of action. Structural studies and biological activities have demonstrated that the myotoxic activity of LYS49-PLA2 is independent of the catalytic activity site. The myotoxic effect is conventionally thought to be to due to the C-terminal region 111–121, which plays an effective role in membrane damage. In the present study, Bn IV LYS49-PLA2 was isolated from Bothrops neuwiedi snake venom in complex with myristic acid (CH3(CH2)12COOH) and its overall structure was refined at 2.2 Å resolution. The Bn IV crystals belong to monoclinic space group P21 and contain a dimer in the asymmetric unit. The unit cell parameters are a = 38.8, b = 70.4, c = 44.0 Å. The biological assembly is a “conventional dimer” and the results confirm that dimer formation is not relevant to the myotoxic activity. Electron density map analysis of the Bn IV structure shows clearly the presence of myristic acid in catalytic site. The relevant structural features for myotoxic activity are located in the C-terminal region and the Bn IV C-terminal residues NKKYRY are a probable heparin binding domain. These findings indicate that the mechanism of interaction between Bn IV and muscle cell membranes is through some kind of cell signal transduction mediated by heparin complexes.

Action of neuwiedase, a metalloproteinase isolated from Bothrops neuwiedi venom, on skeletal muscle: an ultrastructural and immunocytochemistry study

The damaging effects of neuwiedase, a non-hemorrhagic snake venom metalloproteinase from P-I class, on gastrocnemius muscle are studied herein. Following neuwiedase injection, ultrastructural alterations were detected early showing disarrangement of skeletal muscle fibers (characterized by discontinuity of Z lines), mitochondrial swelling, and disruption of plasma membrane and basal lamina. Degradation of skeletal muscle and the appearance of an amorphous substance, primarily composed of cellular debris, were noted after 24 hours. The presence of neuwiedase at the injection site (detected by immunocytochemistry) revealed highly specific labeling of myofibril components of damaged myocytes. In addition, proteolysis of muscle proteins assayed through myofibrils extracted from gastrocnemius muscle indicated that neuwiedase provoked degradation of myofibrils, especially myosin. These results suggest that skeletal muscle damage, induced by neuwiedase, is probably due to its proteolytic action on myofibrils, which are responsible for the maintenance of the cellular architecture.