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1.
J. venom. anim. toxins incl. trop. dis ; 27: e20210024, 2021. tab, graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1340183

Resumo

The Malayan blue coral snake, Calliophis bivirgata flaviceps, is a medically important venomous snake in Southeast Asia. However, the complexity and diversity of its venom genes remain little explored. Methods: To address this, we applied high-throughput next-generation sequencing to profile the venom gland cDNA libraries of C. bivirgata flaviceps. The transcriptome was de novo assembled, followed by gene annotation, multiple sequence alignment and analyses of the transcripts. Results: A total of 74 non-redundant toxin-encoding genes from 16 protein families were identified, with 31 full-length toxin transcripts. Three-finger toxins (3FTx), primarily delta-neurotoxins and cardiotoxin-like/cytotoxin-like proteins, were the most diverse and abundantly expressed. The major 3FTx (Cb_FTX01 and Cb_FTX02) are highly similar to calliotoxin, a delta-neurotoxin previously reported in the venom of C. bivirgata. This study also revealed a conserved tyrosine residue at position 4 of the cardiotoxin-like/cytotoxin-like protein genes in the species. These variants, proposed as Y-type CTX-like proteins, are similar to the H-type CTX from cobras. The substitution is conservative though, preserving a less toxic form of elapid CTX-like protein, as indicated by the lack of venom cytotoxicity in previous laboratory and clinical findings. The ecological role of these toxins, however, remains unclear. The study also uncovered unique transcripts that belong to phospholipase A2 of Groups IA and IB, and snake venom metalloproteinases of PIII subclass, which show sequence variations from those of Asiatic elapids. Conclusion: The venom gland transcriptome of C. bivirgata flaviceps from Malaysia was de novo assembled and annotated. The diversity and expression profile of toxin genes provide insights into the biological and medical importance of the species.(AU)


Assuntos
Animais , Fosfolipases , Mordeduras de Serpentes , Venenos de Víboras/toxicidade , Expressão Gênica , Elapidae/fisiologia
2.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20210024, 2021. tab, graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-33362

Resumo

The Malayan blue coral snake, Calliophis bivirgata flaviceps, is a medically important venomous snake in Southeast Asia. However, the complexity and diversity of its venom genes remain little explored. Methods: To address this, we applied high-throughput next-generation sequencing to profile the venom gland cDNA libraries of C. bivirgata flaviceps. The transcriptome was de novo assembled, followed by gene annotation, multiple sequence alignment and analyses of the transcripts. Results: A total of 74 non-redundant toxin-encoding genes from 16 protein families were identified, with 31 full-length toxin transcripts. Three-finger toxins (3FTx), primarily delta-neurotoxins and cardiotoxin-like/cytotoxin-like proteins, were the most diverse and abundantly expressed. The major 3FTx (Cb_FTX01 and Cb_FTX02) are highly similar to calliotoxin, a delta-neurotoxin previously reported in the venom of C. bivirgata. This study also revealed a conserved tyrosine residue at position 4 of the cardiotoxin-like/cytotoxin-like protein genes in the species. These variants, proposed as Y-type CTX-like proteins, are similar to the H-type CTX from cobras. The substitution is conservative though, preserving a less toxic form of elapid CTX-like protein, as indicated by the lack of venom cytotoxicity in previous laboratory and clinical findings. The ecological role of these toxins, however, remains unclear. The study also uncovered unique transcripts that belong to phospholipase A2 of Groups IA and IB, and snake venom metalloproteinases of PIII subclass, which show sequence variations from those of Asiatic elapids. Conclusion: The venom gland transcriptome of C. bivirgata flaviceps from Malaysia was de novo assembled and annotated. The diversity and expression profile of toxin genes provide insights into the biological and medical importance of the species.(AU)


Assuntos
Animais , Fosfolipases , Mordeduras de Serpentes , Venenos de Víboras/toxicidade , Expressão Gênica , Elapidae/fisiologia
3.
R. bras. Parasitol. Vet. ; 29(2): e002420, 2020. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-27318

Resumo

Hepatozoon pyramidumi sp. n. is described from the blood of the Egyptian saw-scaled viper, Echis pyramidum, captured from Saudi Arabia. Five out of ten viper specimens examined (50%) were found infected with Hepatozoon pyramidumi sp. n. with parasitaemia level ranged from 20-30%. The infection was restricted only to the erythrocytes. Two morphologically different forms of intraerythrocytic stages were observed; small and mature gamonts. The small ganomt with average size of 10.7 × 3.5 μm. Mature gamont was sausage-shaped with recurved poles measuring 16.3 × 4.2 μm in average size. Infected erythrocytes were hypertrophied; their nuclei were deformed and sometimes displaced from their central position in the normal uninfected cell. Merogonic stages were observed in the lung endothelial cell and the liver parenchyma cells. Mature meront was 17.8 × 13.6 µm and contained banana-shaped merozoites with average size of ~15 × 2 µm. Phylogenetic analysis based on the SSU rDNA sequence clustered Hepatozoon pyramidumi sp. n with previously sequenced Hepatozoon spp., most of them infected reptilian hosts without geographic consideration. The morphological and molecular comparison with closely related species proved the taxonomic uniqueness and novelty of the present form.(AU)


Hepatozoon pyramidumi sp. n. é descrito a partir do sangue da víbora em escamas e quilhas serrilhadas, Echis pyramidum, capturada na Arábia Saudita. Cinco de dez espécimes de víbora examinadas (50%) foram encontradas infectadas com Hepatozoon pyramidumi sp. n. com nível de parasitemia de 20% a 30%. A infecção foi restrita apenas aos eritrócitos. Foram observadas duas formas morfologicamente diferentes de estágios intra-eritrocíticos: gamontes de tamanho pequeno e madura. As formas menores de gamontes apresentaram média de 10,7 × 3,5 μm. Os gamontes maduros apresentaram forma de salsicha, com pequenos polos recurvados, medindo 16,3 × 4,2 μm, em média. Os eritrócitos infectados estavam aumentados de tamanho; seus núcleos encontravam-se deformados e, algumas vezes, deslocados de sua posição central, quando comparados às células normais não-infectadas. Foram observados estágios merogônicos em células endoteliais pulmonares e nas células do parênquima hepático. Os merontes maduros apresentavam 17,8 × 13,6 µm e continham merozoítos em forma de banana com tamanho médio de ~ 15 × 2 µm. A análise filogenética baseada nas sequências SSU rDNA agrupou Hepatozoon pyramidumi sp. n com Hepatozoon spp. detectados em répteis de várias regiões geográficas. Por meio de análises morfológicas e moleculares com espécies intimamente relacionadas, demonstrou-se a singularidade dessa nova espécie de Hepatozoon.(AU)


Assuntos
Animais , Elapidae/classificação , Elapidae/genética , Filogenia , Apicomplexa
4.
J. venom. anim. toxins incl. trop. dis ; 26: e20200037, 2020. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135157

Resumo

Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. Methods: The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. Results: The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. Conclusion: This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.(AU)


Assuntos
Animais , Peptídeos , Serina , Venenos de Serpentes , Cisteína Proteases , Elapidae , Peptídeo Hidrolases/isolamento & purificação , Espectrometria de Massas
5.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 26: e20200057, 2020. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-32055

Resumo

Certain environmental toxins permanently damage the thymic epithelium, accelerate immune senescence and trigger secondary immune pathologies. However, the exact underlying cellular mechanisms and pathways of permanent immune intoxication remain unknown. The aim of the present study was to demonstrate gene expressional changes of apoptosis-related cellular pathways in human thymic epithelial cells following exposure to snake venom from Bitis gabonica and Dendroaspis angusticeps. Methods: Snake venoms were characterized by analytical methods including reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, then applied on human thymic epithelial cells (1889c) for 24 h at 10 μg/mL (as used in previous TaqMan Array study). Gene expressional changes restricted to apoptosis were assayed by TaqMan Array (Human Apoptosis Plate). Results: The most prominent gene expressional changes were shown by CASP5 (≈ 2.5 million-fold, confirmed by dedicated quantitative polymerase chain reaction) and CARD9 (0.016-fold) for B. gabonica, and BIRC7 (6.46-fold) and CASP1 (0.30-fold) for D. angusticeps. Conclusion: The observed apoptotic environment suggests that pyroptosis may be the dominant pathway through which B. gabonica and D. angusticeps snake venoms trigger thymic epithelial apoptosis following envenomation.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Venenos de Serpentes/genética , Apoptose/genética , Células Epiteliais , Piroptose , Viperidae , Elapidae
6.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 26: e20200037, 2020. graf
Artigo em Inglês | VETINDEX | ID: vti-32312

Resumo

Proteases play an important role for the proper physiological functions of the most diverse organisms. When unregulated, they are associated with several pathologies. Therefore, proteases have become potential therapeutic targets regarding the search for inhibitors. Snake venoms are complex mixtures of molecules that can feature a variety of functions, including peptidase inhibition. Considering this, the present study reports the purification and characterization of a Kunitz-type peptide present in the Dendroaspis polylepis venom as a simultaneous inhibitor of elastase-1 and cathepsin L. Methods: The low molecular weight pool from D. polylepis venom was fractionated in reverse phase HPLC and all peaks were tested in fluorimetric assays. The selected fraction that presented inhibitory activity over both proteases was submitted to mass spectrometry analysis, and the obtained sequence was determined as a Kunitz-type serine protease inhibitor homolog dendrotoxin I. The molecular docking of the Kunitz peptide on the elastase was carried out in the program Z-DOCK, and the program RosettaDock was used to add hydrogens to the models, which were re-ranked using ZRANK program. Results: The fraction containing the Kunitz molecule presented similar inhibition of both elastase-1 and cathepsin L. This Kunitz-type peptide was characterized as an uncompetitive inhibitor for elastase-1, presenting an inhibition constant (Ki) of 8 μM. The docking analysis led us to synthesize two peptides: PEP1, which was substrate for both elastase-1 and cathepsin L, and PEP2, a 30-mer cyclic peptide, which showed to be a cathepsin L competitive inhibitor, with a Ki of 1.96 µM, and an elastase-1 substrate. Conclusion: This work describes a Kunitz-type peptide toxin presenting inhibitory potential over serine and cysteine proteases, and this could contribute to further understand the envenomation process by D. polylepis. In addition, the PEP2 inhibits the cathepsin L activity with a low inhibition constant.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Venenos de Serpentes/síntese química , Peptídeos/isolamento & purificação , Inibidores de Proteases/análise , Elapidae , Serina/biossíntese , Serina/isolamento & purificação , Cisteína/biossíntese , Cisteína/isolamento & purificação
7.
J. venom. anim. toxins incl. trop. dis ; 25: e143018, 2019. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1002498

Resumo

This paper aims to highlight and analyze discrepancies in reporting of deaths due to venomous animals in Brazil, from 2001 to 2015, between two national information systems: The Notifiable Diseases Information System (Sistema de Informação de Agravos de Notificação - SINAN) and the Mortality Information System (Sistema de Informações sobre Mortalidade - SIM). Methods: Descriptive and comparative study of the SINAN and SIM information systems, was conducted via the following steps: collecting the death notices from SINAN and SIM; constructing tables and comparative graphics; and, only in scorpion sting fatalities, analyzing the distribution of deaths by age group as described in the specialized literature. Results: While SINAN identifies strong growth in the number of deaths from scorpion stings, SIM shows greater increase in the number of reported deaths from bee stings, especially in the South and Southeast regions. Notably, bees are the sole etiological agent that received more reports in SIM than in SINAN for every year in the period studied. The age-group distribution of the data on deaths from scorpion stings reinforced the indication of problems occurring in their registration in SINAN, especially since 2007, which may have an effect on analyses based on these data. Conclusion: Comparative analysis of these databases permits identification of important differences between profiles presented by these systems, which have equal relevance for Brazil as a whole and for its regions. These differences may influence the construction of various scenarios.(AU)


Assuntos
Animais , Escorpiões , Abelhas , Mordeduras e Picadas , Sistemas de Informação , Picadas de Escorpião , Animais Peçonhentos , Atestado de Óbito , Elapidae
8.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 1-5, 2018. tab, mapas, ilus
Artigo em Inglês | VETINDEX | ID: vti-732652

Resumo

Background Bites provoked by the genus Micrurus represent less than 1% of snakebite cases notified in Brazil, a tiny fraction compared with other genus such as Bothrops and Crotalus, which together represent almost 80% of accidents. In addition to their less aggressive behavior, habits and morphology of coral snakes are determinant factors for such low incidence of accidents. Although Micrurus bites are rare, victims must be rescued and hospitalized in a short period of time, because this type of envenoming may evolve to a progressive muscle weakness and acute respiratory failure. Case Presentation We report an accident caused by Micrurus corallinus involving a 28-year-old Caucasian sailor man bitten on the hand. The accident occurred in a recreational camp because people believed the snake was not venomous. The victim presented neurological symptoms 2 h after the accident and was taken to the hospital, where he received antielapidic serum 10 h after the bite. After the antivenom treatment, the patient presented clinical evolution without complications and was discharged 4 days later. Conclusions We reinforce that it is essential to have a health care structure suitable for the treatment of snakebite. Besides, the manipulation of these animals should only be carried out by a team of well-equipped and trained professionals, and even so with special attention.(AU)


Assuntos
Humanos , Animais , Elapidae , Venenos Elapídicos/intoxicação , Mordeduras de Serpentes/complicações , Intoxicação , Brasil , Venenos/efeitos adversos
9.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 41, Jan. 24, 2018. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18764

Resumo

Background: The oral cavities of snakes are replete with various types of bacterial flora. Culture-dependent studies suggest that some of the bacterial species are responsible for secondary bacterial infection associated with snakebite. A complete profile of the ophidian oral bacterial community has been unreported until now. Therefore, in the present study, we determined the complete bacterial compositions in the oral cavity of some snakes from India. Methods: Total DNA was isolated from oral swabs collected from three wild snake species (Indian Cobra, King Cobra and Indian Python). Next, the DNA was subjected to PCR amplification of microbial 16S rRNA gene using V3-region-specific primers. The amplicons were used for preparation of DNA libraries that were sequenced on an Illumina MiSeq platform. Results: The cluster-based taxonomy analysis revealed that Proteobacteria and Actinobacteria were the most predominant phyla present in the oral cavities of snakes. This result indicates that snakes show more similarities to birds than mammals as to their oral bacterial communities. Furthermore, our study reports all the unique and common bacterial species (total: 147) found among the oral microbes of snakes studied, while the majority of commonly abundant species were pathogens or opportunistic pathogens to humans. A wide difference in ophidian oral bacterial flora suggests variation by individual, species and geographical region. Conclusion: The present study would provide a foundation for further research on snakes to recognize the potential drugs/antibiotics for the different infectious diseases.(AU)


Assuntos
Animais , Análise de Sequência de RNA/veterinária , Boca/microbiologia , Microbiota , Elapidae/microbiologia , Mordeduras de Serpentes/complicações , Mordeduras de Serpentes/microbiologia , Índia
10.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 1-11, 2018. graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-18170

Resumo

Background Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Methods Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Results Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4- C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. Conclusions This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.(AU)


Assuntos
Humanos , Animais , Venenos Elapídicos/isolamento & purificação , Elapidae , Motilidade dos Espermatozoides , Fármacos para a Fertilidade Masculina , Sêmen , Reações Bioquímicas , Espectrometria de Massas em Tandem/métodos
11.
J. venom. anim. toxins incl. trop. dis ; 24: 1-11, 2018. graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484741

Resumo

Background Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Methods Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Results Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4- C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. Conclusions This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.


Assuntos
Humanos , Animais , Elapidae , Fármacos para a Fertilidade Masculina , Motilidade dos Espermatozoides , Sêmen , Venenos Elapídicos/isolamento & purificação , Espectrometria de Massas em Tandem/métodos , Reações Bioquímicas
12.
J. venom. anim. toxins incl. trop. dis ; 24: 1-5, 2018. tab, map, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484747

Resumo

Background Bites provoked by the genus Micrurus represent less than 1% of snakebite cases notified in Brazil, a tiny fraction compared with other genus such as Bothrops and Crotalus, which together represent almost 80% of accidents. In addition to their less aggressive behavior, habits and morphology of coral snakes are determinant factors for such low incidence of accidents. Although Micrurus bites are rare, victims must be rescued and hospitalized in a short period of time, because this type of envenoming may evolve to a progressive muscle weakness and acute respiratory failure. Case Presentation We report an accident caused by Micrurus corallinus involving a 28-year-old Caucasian sailor man bitten on the hand. The accident occurred in a recreational camp because people believed the snake was not venomous. The victim presented neurological symptoms 2 h after the accident and was taken to the hospital, where he received antielapidic serum 10 h after the bite. After the antivenom treatment, the patient presented clinical evolution without complications and was discharged 4 days later. Conclusions We reinforce that it is essential to have a health care structure suitable for the treatment of snakebite. Besides, the manipulation of these animals should only be carried out by a team of well-equipped and trained professionals, and even so with special attention.


Assuntos
Humanos , Animais , Elapidae , Intoxicação , Mordeduras de Serpentes/complicações , Venenos Elapídicos/intoxicação , Brasil , Venenos/efeitos adversos
13.
Artigo em Inglês | VETINDEX | ID: vti-18643

Resumo

It is of popular and scientific knowledge that toxins from snake venom (among them the PLA2 and myotoxins) are neutralized by various compounds, such as antibodies and proteins purified from animal blood. Venomous and nonvenomous snakes have PLA2 inhibitory proteins, called PLIs, in their blood serum. One hypothesis that could explain the presence of these PLIs in the serum of venomous snakes would be self-protection against the enzymes of their own venom, which eventually could reach the circulatory system. However, the presence of PLIs in non-venomous snakes suggests that their physiological role might not be restricted to protection against PLA2 toxins, but could be extended to other functions, as in the innate immune system and local regulation of PLA2s. The present study aimed to review the currently available literature on PLA2 and myotoxin alpha inhibitors present in snake plasma, thus helping to improve the research on these molecules. Furthermore, this review includes current information regarding the mechanism of action of these inhibitors in an attempt to better understand their application, and proposes the use of these molecules as new models in snakebite therapy. These molecules may help in the neutralization of different types of phospholipases A2 and myotoxins, complementing the conventional serum therapy.(AU)


Assuntos
Animais , Elapidae/sangue , Fosfolipases A2 , Trimeresurus/fisiologia
14.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954790

Resumo

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.(AU)


Assuntos
Dobramento de Proteína , Elapidae , Venenos Elapídicos , Anticorpos , Neurotoxinas
15.
Cad. técn. Vet. Zoot. ; (75): 39-41, dez. 2014. tab, ilus
Artigo em Português | VETINDEX | ID: vti-481334

Resumo

O acidente elapídicoé causado por serpentes da família Elapidae,gêneros Micrurus e Leptomicrurus. No Brasil, há 19 espécies de Micrurus e três de Leptomicrurus (Quadro 1). As espécies mais importantes são Micrurusfrontalis (cobra-coral,coral), Micrurus corallinus(cobra-coral, coral--verdadeira, boicorá)e Micrurus ibiboboca(coral-venenosa, coral--verdadeira). São serpentes de tamanho mediano(300-1.200mm),que apresentam anéis corporais de colorações pretas, amarelas, brancas e vermelhas. Os olhos são pequenos com pupilas arredondadas.Possuem dentição proteróglifa, então têm fosseta loreal. Uma característica marcante é o comportamento de levantarem a cauda, de forma semienrolada (engodocaudal), quando se sentem ameaçadas.


Assuntos
Animais , Elapidae/anatomia & histologia , Elapidae/fisiologia , Mordeduras de Serpentes/patologia , Animais Peçonhentos/anatomia & histologia , Animais Peçonhentos/fisiologia , Sorologia/métodos , Mordeduras de Serpentes/diagnóstico , Peçonhas/efeitos adversos
17.
Artigo em Inglês | VETINDEX | ID: vti-9289

Resumo

In Guinea Elapids are responsible for 20% of envenomations. The associated case fatality rate (CFR) ranged 15-27%, irrespective of treatment. Results We studied 77 neurotoxic envenomations divided in 3 groups: a set of patients that received only traditional or symptomatic treatments, and two other groups that received either 2 or 4 initial vials of Antivipmyn® Africa renewed as necessary. CFR was 27.3%, 15.4% and 17.6%, respectively. Although antivenom treatment was likely to reduce CFR, it didnt seem to have an obvious clinical benefit for the patients, suggesting a low treatment efficacy. Mean delay to treatment or clinical stages were not significantly different between the patients who recovered and the patients who died, or between groups. Interpretation of these results is complicated by the lack of systematic studies under comparable conditions. Of particular importance is the absence of assisted ventilation, available to patients in all the other clinical studies of neurotoxic envenomation. Conclusion The apparent lack of clinical benefit may have several causes. The hypothesis of a limited therapeutic window, i.e. an insufficient formation of antigen-antibody complexes once toxins are bound to their targets and/or distributed beyond the reach of antivenom, should be explored.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Intoxicação/complicações , Antivenenos/análise , Neurotoxinas , Serpentes
18.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484525

Resumo

In Guinea Elapids are responsible for 20% of envenomations. The associated case fatality rate (CFR) ranged 15-27%, irrespective of treatment. Results We studied 77 neurotoxic envenomations divided in 3 groups: a set of patients that received only traditional or symptomatic treatments, and two other groups that received either 2 or 4 initial vials of Antivipmyn® Africa renewed as necessary. CFR was 27.3%, 15.4% and 17.6%, respectively. Although antivenom treatment was likely to reduce CFR, it didn’t seem to have an obvious clinical benefit for the patients, suggesting a low treatment efficacy. Mean delay to treatment or clinical stages were not significantly different between the patients who recovered and the patients who died, or between groups. Interpretation of these results is complicated by the lack of systematic studies under comparable conditions. Of particular importance is the absence of assisted ventilation, available to patients in all the other clinical studies of neurotoxic envenomation. Conclusion The apparent lack of clinical benefit may have several causes. The hypothesis of a limited therapeutic window, i.e. an insufficient formation of antigen-antibody complexes once toxins are bound to their targets and/or distributed beyond the reach of antivenom, should be explored.


Assuntos
Animais , Antivenenos/análise , Intoxicação/complicações , Neurotoxinas , Venenos de Serpentes/análise , Serpentes
19.
J. venom. anim. toxins incl. trop. dis ; 18(2): 150-156, 2012. graf, tab
Artigo em Inglês | LILACS, VETINDEX | ID: lil-639473

Resumo

Cytotoxicity of venoms from eleven medically important snakes found in Southeast Asia (Naja kaouthia, Naja siamensis, Naja sumatrana, Ophiophagus hannah, Bungarus candidus, Bungarus fasciatus, Enhydrina schistosa, Calloselasma rhodostoma, Trimeresurus purpureomaculatus and Tropidolaemus sumatranus) was determined, based on the MTS cytotoxicity assay, which determines the survival of viable cells in monolayer MDCK and Vero cell cultures upon exposure to the snake venoms. Snake venom toxicity was expressed as the venom dose that killed 50% of the cells (CTC50) under the assay conditions. Venoms of C. rhodostoma (2.6 µg/mL, 1.4 µg/mL) and O. hannah were the most cytotoxic (3.8 µg/mL, 1.7 µg/mL) whereas N. siamensis venom showed the least cytotoxicity (51.9 µg/mL, 45.7 µg/mL) against Vero and MDCK cells, respectively. All the viper venoms showed higher cytotoxic potency towards both Vero and MDCK cell lines, in comparison to krait and cobra venoms. E. schistosa did not cause cytotoxicity towards MDCK or Vero cells at the tested concentrations. The cytotoxicity correlates well with the known differences in the composition of venoms from cobras, kraits, vipers and sea snakes.(AU)


Assuntos
Animais , Venenos de Serpentes , Elapidae , Citotoxicidade Imunológica , Elapidae , Naja naja
20.
Acta amaz. ; 42(1)2012.
Artigo em Português | VETINDEX | ID: vti-450720

Resumo

Despite the importance of accidents involving snakebites, research on this theme in Brazil is relatively rare, especially in the Amazon region. Due to the poor number of studies about ophidism in the state of Acre and to the importance of this type of research in public health, epidemiological works on snake bite accidents are of great relevance. This study presents the list of species of poisonous snakes and the epidemiological aspects of snakebites in Cruzeiro do Sul, region of Alto Juruá (State of Acre, Brazil), in order to verify the snake genera responsible for the bites, and the main aspects involving the accidents and hospital attending. The epidemiologic data were collected from SINAN (National Databank of Major Causes of Morbidity) in the sector of epidemiologic vigilance of Hospital Regional do Juruá, in Cruzeiro do Sul. Seven species of poisonous snakes were recorded in Cruzeiro do Sul: three viperids (Bothrops atrox, Bothriopsis bilineata and Lachesis muta) and four elapids (Micrurus hemprichii, M. lemniscatus, M. remotus and M. surinamensis). During a two-year period (August 2007 to July 2009) 195 cases of snakebites were recorded. Fifty-one percent of the accidents were classified as lachetic (Lachesis), followed by bothropic (Bothrops and Bothriopsis) with 38% and crotalic (Crotalus) with 2%. In 9% of cases the snake genus involved was not informed. The majority of the accidents involved adult males living in rural areas, mainly affected in the lower limbs. The cases occurred most frequently from November to April, coinciding with the highest pluviometric levels. Most snakebites were equivocally attributed to L. muta, and were probably caused by B. atrox.


Apesar da importância dos acidentes ofídicos na Saúde Pública, são relativamente poucas as pesquisas realizadas sobre esse tema no Brasil. Devido aos poucos estudos sobre ofidismo na Amazônia e especialmente no estado do Acre, trabalhos epidemiológicos são de grande relevância. Esse estudo apresenta a lista de serpentes peçonhentas e aspectos epidemiológicos dos acidentes ofídicos em Cruzeiro do Sul, região do Alto Juruá (Acre), verificando quais gêneros de serpentes são responsáveis pelos envenenamentos e aspectos que envolvem o acidente e o atendimento hospitalar. Os dados epidemiológicos foram coligidos a partir do SINAN (Sistema de Informações de Agravos de Notificação), no setor de vigilância epidemiológica do Hospital Regional do Juruá, no município de Cruzeiro do Sul. Sete espécies de serpentes peçonhentas foram registradas nesse município: três viperídeos (Bothrops atrox, Bothriopsis bilineata e Lachesis muta) e quatro elapídeos (Micrurus hemprichii, M. lemniscatus, M. remotus and M. surinamensis). Durante o período de dois anos (agosto de 2007 a julho de 2009) foram registrados 195 casos de acidentes ofídicos. Cinquenta e um por cento dos acidentes foi classificado como laquético (Lachesis), seguido pelo botrópico (Bothrops e Bothriopsis) com 38% e crotálico (Crotalus) com 2%. Em 9% dos casos o gênero da serpente envolvida não foi informado. A maioria dos acidentes envolveu indivíduos adultos do gênero masculino em área rural, afetados principalmente nos membros inferiores. Os casos ocorreram mais frequentemente nos meses de novembro a abril, coincidindo com os maiores níveis pluviométricos. A maioria dos acidentes foi atribuída equivocadamente à serpente L. muta, tendo sido provavelmente causados por B. atrox.

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