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1.
J. venom. anim. toxins incl. trop. dis ; 28: e20220026, 2022. tab, graf, ilus
Artigo em Inglês | VETINDEX | ID: biblio-1418297

Resumo

Background: Scorpion neurotoxins such as those that modify the mammalian voltagegated 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.(AU)


Assuntos
Animais , Venenos de Escorpião/enzimologia , Neurotoxinas/análise , Proteínas Recombinantes/análise
2.
Acta sci. vet. (Impr.) ; 50(suppl.1): Pub.753-4 jan. 2022. ilus
Artigo em Português | VETINDEX | ID: biblio-1458561

Resumo

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...


Assuntos
Animais , Botulismo/epidemiologia , Botulismo/veterinária , Clostridium botulinum tipo C/isolamento & purificação , Galinhas/microbiologia , Neurotoxinas , Brasil , Surtos de Doenças/veterinária
3.
Acta sci. vet. (Online) ; 50(suppl.1): Pub. 753, 15 fev. 2022. ilus
Artigo em Português | VETINDEX | ID: vti-765212

Resumo

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...(AU)


Assuntos
Animais , Botulismo/epidemiologia , Botulismo/veterinária , Clostridium botulinum tipo C/isolamento & purificação , Neurotoxinas , Galinhas/microbiologia , Brasil , Surtos de Doenças/veterinária
4.
J. venom. anim. toxins incl. trop. dis ; 28: e20210094, 2022. graf, tab, ilus
Artigo em Inglês | VETINDEX | ID: biblio-1395948

Resumo

Background: Endogenous phospholipases A2 (PLA2 ) play a fundamental role in inflammation, neurodegenerative diseases, apoptosis and cellular senescence. Neurotoxins with PLA2 activity are found in snake venoms from the Elapidae and Viperidae families. The mechanism of action of these neurotoxins have been studied using hippocampal and cerebellar neuronal cultures showing [Ca2+]i increase, mitochondrial depolarization and cell death. Astrocytes are rarely used as a model, despite being modulators at the synapses and responsible for homeostasis and defense in the central nervous system. Preserving the cell division ability, they can be utilized to study the cell proliferation process. In the present work cultured astrocytes and glioblastoma cells were employed to characterize the action of ß-micrustoxin (previously named Mlx-9), a PLA2 isolated from Micrurus lemniscatus snake venom. The ß-micrustoxin structure was determined and the cell proliferation, cell cycle phases and the regulatory proteins p53, p21 and p27 were investigated. Methods: ß-micrustoxin was characterized biochemically by a proteomic approach. Astrocytes were obtained by dissociation of pineal glands from Wistar rats; glioblastoma tumor cells were purchased from ATCC and Sigma and cultured in DMEM médium. Cell viability was evaluated by MTT assay; cell proliferation and cell cycle phases were analyzed by flow cytometry; p53, p21 and p27 proteins were studied by western blotting and immunocytochemistry. Results: Proteomic analysis revealed fragments on ß-micrustoxin that aligned with a PLA2 from Micrurus lemniscatus lemniscatus previously identified as transcript ID DN112835_C3_g9_i1/m.9019. ß-micrustoxin impaired the viability of astrocytes and glioblastoma tumor cells. There was a reduction in cell proliferation, an increase in G2/M phase and activation of p53, p21 and p27 proteins in astrocytes. Conclusion: These findings indicate that ß-micrustoxin from Micrurus lemniscatus venom could inhibit cell proliferation through p53, p21 and p27 activation thus imposing cell cycle arrest at the checkpoint G2/M.(AU)


Assuntos
Venenos de Serpentes/toxicidade , Bioquímica , Glioblastoma , Neurotoxinas
5.
J. venom. anim. toxins incl. trop. dis ; 27: e20210023, 2021. tab, graf, ilus, mapas
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1346437

Resumo

Diverse and unique bioactive neurotoxins known as conopeptides or conotoxins are produced by venomous marine cone snails. Currently, these small and stable molecules are of great importance as research tools and platforms for discovering new drugs and therapeutics. Therefore, the characterization of Conus venom is of great significance, especially for poorly studied species. Methods: In this study, we used bioanalytical techniques to determine the venom profile and emphasize the functional composition of conopeptides in Conus taeniatus, a neglected worm-hunting cone snail. Results: The proteomic analysis revealed that 84.0% of the venom proteins were between 500 and 4,000 Da, and 16.0% were > 4,000 Da. In C. taeniatus venom, 234 peptide fragments were identified and classified as conotoxin precursors or non-conotoxin proteins. In this process, 153 conotoxin precursors were identified and matched to 23 conotoxin precursors and hormone superfamilies. Notably, the four conotoxin superfamilies T (22.87%), O1 (17.65%), M (13.1%) and O2 (9.8%) were the most abundant peptides in C. taeniatus venom, accounting for 63.40% of the total conotoxin diversity. On the other hand, 48 non-conotoxin proteins were identified in the venom of C. taeniatus. Moreover, several possibly biologically active peptide matches were identified, and putative applications of the peptides were assigned. Conclusion: Our study showed that the composition of the C. taeniatus-derived proteome is comparable to that of other Conus species and contains an effective mix of toxins, ionic channel inhibitors and antimicrobials. Additionally, it provides a guidepost for identifying novel conopeptides from the venom of C. taeniatus and discovering conopeptides of potential pharmaceutical importance.(AU)


Assuntos
Animais , Proteoma , Conotoxinas , Caramujo Conus , Venenos de Moluscos , Neurotoxinas , Produtos Biológicos
6.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20210023, 2021. tab, graf, ilus, mapas
Artigo em Inglês | VETINDEX | ID: vti-31889

Resumo

Diverse and unique bioactive neurotoxins known as conopeptides or conotoxins are produced by venomous marine cone snails. Currently, these small and stable molecules are of great importance as research tools and platforms for discovering new drugs and therapeutics. Therefore, the characterization of Conus venom is of great significance, especially for poorly studied species. Methods: In this study, we used bioanalytical techniques to determine the venom profile and emphasize the functional composition of conopeptides in Conus taeniatus, a neglected worm-hunting cone snail. Results: The proteomic analysis revealed that 84.0% of the venom proteins were between 500 and 4,000 Da, and 16.0% were > 4,000 Da. In C. taeniatus venom, 234 peptide fragments were identified and classified as conotoxin precursors or non-conotoxin proteins. In this process, 153 conotoxin precursors were identified and matched to 23 conotoxin precursors and hormone superfamilies. Notably, the four conotoxin superfamilies T (22.87%), O1 (17.65%), M (13.1%) and O2 (9.8%) were the most abundant peptides in C. taeniatus venom, accounting for 63.40% of the total conotoxin diversity. On the other hand, 48 non-conotoxin proteins were identified in the venom of C. taeniatus. Moreover, several possibly biologically active peptide matches were identified, and putative applications of the peptides were assigned. Conclusion: Our study showed that the composition of the C. taeniatus-derived proteome is comparable to that of other Conus species and contains an effective mix of toxins, ionic channel inhibitors and antimicrobials. Additionally, it provides a guidepost for identifying novel conopeptides from the venom of C. taeniatus and discovering conopeptides of potential pharmaceutical importance.(AU)


Assuntos
Animais , Proteoma , Conotoxinas , Caramujo Conus , Venenos de Moluscos , Neurotoxinas , Produtos Biológicos
7.
J. venom. anim. toxins incl. trop. dis ; 27: e20200196, 2021. tab, graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1346436

Resumo

Snake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey. In the present study, in order to unravel the proteomic repertoire of Deinagkistrodon acutus venom, some trace abundance components were analyzed. Methods Shotgun proteomic approach combined with shotgun nano-LC-ESI-MS/MS were employed to characterize the medically important D. acutus venom, after collected samples were enriched with the combinatorial peptide ligand library (CPLL). Results This avenue helped us find some trace components, undetected before, in D. acutus venom. The results indicated that D. acutus venom comprised 84 distinct proteins from 10 toxin families and 12 other proteins. These results are more than twice the number of venom components obtained from previous studies, which were only 29 distinct proteins obtained through RP-HPLC for the venom of the same species. The present results indicated that in D. acutus venom, the most abundant components (66.9%) included metalloproteinases, serine proteinases, and C-type lectin proteins; the medium abundant components (13%) comprised phospholipases A2 (PLA2) and 5'-nucleotidases and nucleases; whereas least abundant components (6%) were aminopeptidases, L-amino acid oxidases (LAAO), neurotoxins and disintegrins; and the trace components. The last were undetected before the use of conventional shotgun proteomics combined with shotgun nano-LC-ESI-MS/MS, such as cysteine-rich secretory proteins Da-CRPa, phospholipases B-like 1, phospholipases B (PLB), nerve growth factors (NGF), glutaminyl-peptide cyclortransferases (QC), and vascular non-inflammatory molecules 2 (VNN2). Conclusion These findings demonstrated that the CPLL enrichment method worked well in finding the trace toxin proteins in D. acutus venom, in contrast with the previous venomic characterization of D. acutus by conventional LC-MS/MS. In conclusion, this approach combined with the CPLL enrichment was effective for allowing us to explore the hidden D. acutus venomic profile and extended the list of potential venom toxins.(AU)


Assuntos
Animais , Oxirredutases , Peptídeos , Venenos de Víboras , Proteoma , Neurotoxinas
8.
J. venom. anim. toxins incl. trop. dis ; 27: e20200173, 2021. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1279403

Resumo

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)


Assuntos
Animais , Venenos de Escorpião , Sódio/análise , Biologia Computacional , Neurotoxinas
9.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20200173, 2021. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-31951

Resumo

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)


Assuntos
Animais , Venenos de Escorpião , Sódio/análise , Biologia Computacional , Neurotoxinas
10.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20200196, 2021. tab, graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-31887

Resumo

Snake venoms are complex mixtures of toxic proteins or peptides encoded by various gene families that function synergistically to incapacitate prey. In the present study, in order to unravel the proteomic repertoire of Deinagkistrodon acutus venom, some trace abundance components were analyzed. Methods Shotgun proteomic approach combined with shotgun nano-LC-ESI-MS/MS were employed to characterize the medically important D. acutus venom, after collected samples were enriched with the combinatorial peptide ligand library (CPLL). Results This avenue helped us find some trace components, undetected before, in D. acutus venom. The results indicated that D. acutus venom comprised 84 distinct proteins from 10 toxin families and 12 other proteins. These results are more than twice the number of venom components obtained from previous studies, which were only 29 distinct proteins obtained through RP-HPLC for the venom of the same species. The present results indicated that in D. acutus venom, the most abundant components (66.9%) included metalloproteinases, serine proteinases, and C-type lectin proteins; the medium abundant components (13%) comprised phospholipases A2 (PLA2) and 5'-nucleotidases and nucleases; whereas least abundant components (6%) were aminopeptidases, L-amino acid oxidases (LAAO), neurotoxins and disintegrins; and the trace components. The last were undetected before the use of conventional shotgun proteomics combined with shotgun nano-LC-ESI-MS/MS, such as cysteine-rich secretory proteins Da-CRPa, phospholipases B-like 1, phospholipases B (PLB), nerve growth factors (NGF), glutaminyl-peptide cyclortransferases (QC), and vascular non-inflammatory molecules 2 (VNN2). Conclusion These findings demonstrated that the CPLL enrichment method worked well in finding the trace toxin proteins in D. acutus venom, in contrast with the previous venomic characterization of D. acutus by conventional LC-MS/MS. In conclusion, this approach combined with the CPLL enrichment was effective for allowing us to explore the hidden D. acutus venomic profile and extended the list of potential venom toxins.(AU)


Assuntos
Animais , Oxirredutases , Peptídeos , Venenos de Víboras , Proteoma , Neurotoxinas
11.
J. venom. anim. toxins incl. trop. dis ; 27: e20210001, 2021. tab, graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1351017

Resumo

Phα1ß is a neurotoxin purified from spider venom that acts as a high-voltage-activated (HVA) calcium channel blocker. This spider peptide has shown a high selectivity for N-type HVA calcium channels (NVACC) and an analgesic effect in several animal models of pain. Its activity was associated with a reduction in calcium transients, glutamate release, and reactive oxygen species production from the spinal cord tissue and dorsal ganglia root (DRG) in rats and mice. It has been reported that intrathecal (i.t.) administration of Phα1ß to treat chronic pain reverted opioid tolerance with a safer profile than ω-conotoxin MVIIA, a highly selective NVACC blocker. Following a recent development of recombinant Phα1ß (CTK 01512-2), a new molecular target, TRPA1, the structural arrangement of disulphide bridges, and an effect on glial plasticity have been identified. CTK 01512-2 reproduced the antinociceptive effects of the native toxin not only after the intrathecal but also after the intravenous administration. Herein, we review the Phα1ß antinociceptive activity in the most relevant pain models and its mechanisms of action, highlighting the impact of CTK 01512-2 synthesis and its potential for multimodal analgesia.


Assuntos
Dor , Peptídeos/isolamento & purificação , Espécies Reativas de Oxigênio , Analgésicos/efeitos adversos , Neurotoxinas/isolamento & purificação
12.
J. venom. anim. toxins incl. trop. dis ; 27: e20210001, 2021. tab, graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484769

Resumo

Phα1ß is a neurotoxin purified from spider venom that acts as a high-voltage-activated (HVA) calcium channel blocker. This spider peptide has shown a high selectivity for N-type HVA calcium channels (NVACC) and an analgesic effect in several animal models of pain. Its activity was associated with a reduction in calcium transients, glutamate release, and reactive oxygen species production from the spinal cord tissue and dorsal ganglia root (DRG) in rats and mice. It has been reported that intrathecal (i.t.) administration of Phα1ß to treat chronic pain reverted opioid tolerance with a safer profile than ω-conotoxin MVIIA, a highly selective NVACC blocker. Following a recent development of recombinant Phα1ß (CTK 01512-2), a new molecular target, TRPA1, the structural arrangement of disulphide bridges, and an effect on glial plasticity have been identified. CTK 01512-2 reproduced the antinociceptive effects of the native toxin not only after the intrathecal but also after the intravenous administration. Herein, we review the Phα1ß antinociceptive activity in the most relevant pain models and its mechanisms of action, highlighting the impact of CTK 01512-2 synthesis and its potential for multimodal analgesia.


Assuntos
Analgésicos/efeitos adversos , Dor , Espécies Reativas de Oxigênio , Neurotoxinas/isolamento & purificação , Peptídeos/isolamento & purificação
13.
J. venom. anim. toxins incl. trop. dis ; 25: e148218, 2019. graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1002496

Resumo

Tityus serrulatus venom (Ts venom) is a complex mixture of several compounds with biotechnological and therapeutical potentials, which highlights the importance of the identification and characterization of these components. Although a considerable number of studies have been dedicated to the characterization of this complex cocktail, there is still a limitation of knowledge concerning its venom composition. Most of Ts venom studies aim to isolate and characterize their neurotoxins, which are small, basic proteins and are eluted with high buffer concentrations on cation exchange chromatography. The first and largest fraction from carboxymethyl cellulose-52 (CMC-52) chromatography of Ts venom, named fraction I (Fr I), is a mixture of proteins of high and low molecular masses, which do not interact with the cation exchange resin, being therefore a probable source of components still unknown of this venom. Thus, the present study aimed to perform the proteome study of Fraction I from Ts venom, by high resolution mass spectrometry, and its biochemical characterization, by the determination of several enzymatic activities. Methods: Fraction I was obtained by a cation exchange chromatography using 50 mg of crude venom. This fraction was subjected to a biochemical characterization, including determination of L-amino acid oxidase, phospholipase, hyaluronidase, proteases activities and inhibition of angiotensin converting enzyme (ACE) activity. Fraction I was submitted to reduction, alkylation and digestion processes, and the tryptic digested peptides obtained were analyzed in a Q-Exactive Orbitrap mass spectrometer. Data analysis was performed by PEAKS 8.5 software against NCBI database. Results: Fraction I exhibits proteolytic activity and it was able to inhibit ACE activity. Its proteome analysis identified 8 different classes of venom components, among them: neurotoxins (48%), metalloproteinases (21%), hypotensive peptides (11%), cysteine-rich venom protein (9%), antimicrobial peptides (AMP), phospholipases and other enzymes (chymotrypsin and lysozymes) (3%) and phosphodiesterases (2%). Conclusions: The combination of a proteomic and biochemical characterization strategies leads us to identify new components in the T. serrulatus scorpion venom. The proteome of venom´s fraction can provide valuable direction in the obtainment of components in their native forms in order to perform a preliminary characterization and, consequently, to promote advances in biological discoveries in toxinology.(AU)


Assuntos
Animais , Venenos de Escorpião , Produtos Biológicos , Proteoma , Metaloproteases , Neurotoxinas , Fosfolipases , Enzimas
14.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 25: e148818, May 16, 2019. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-19838

Resumo

Background: L-Glutamate (L-Glu), the major excitatory neurotransmitter in the mammalian Central Nervous System (CNS), is essential to cognitive functions. However, when L-Glu is accumulated in large concentrations at the synaptic cleft, it can induce excitotoxicity that results in secondary damage implicated in many neurological disorders. Current therapies for the treatment of neurological disorders are ineffective and have side effects associated with their use; therefore, there is a need to develop novel treatments. In this regard, previous studies have shown that neuroactive compounds obtained from the venom of the spider Parawixia bistriata have neuroprotective effects in vitro and in vivo. In this sense, this work aimed to evaluate potential neuroprotective effects of fraction RT10, obtained from this spider venom, on primary cultures of neuron and glial cells subjected to glutamate excitotoxicity insults. Methods: Primary cultures of neurons and glia were obtained from the cerebral tissue of 1-day-old postnatal Wistar rats. After 7 days in vitro (DIV), the cultures were incubated with fraction RT10 (0.002; 0.02; 0.2 and 2 µg/µL) or riluzole (100 µM) for 3-hours before application of 5 mM L-Glu. After 12 hours, the resazurin sodium salt (RSS) test was applied to measure metabolic activity and proliferation of living cells, whereas immunocytochemistry for MAP2 was performed to measure neuronal survival. In addition, the cells were immunolabeled with NeuN and GFAP in baseline conditions. Results: In the RSS tests, we observed that pre-incubation with RT10 before the excitotoxic insults from L-Glu resulted in neuroprotection, shown by a 10% reduction in the cell death level. RT10 was more effective than riluzole, which resulted in a cell-death reduction of 5%. Moreover, qualitative analysis of neuronal morphology (by MAP2 staining, expressed as fluorescence intensity (FI), an indirect measure of neuronal survival)...(AU)


Assuntos
Animais , Ratos , Ratos Wistar , Neuroglia/química , Glutamatos/análise , Glutamatos/toxicidade , Venenos de Aranha/uso terapêutico , Fármacos Neuroprotetores/administração & dosagem , Neurotoxinas/análise
15.
Acta sci. vet. (Impr.) ; 46(supl): 1-3, 2018. ilus
Artigo em Inglês | VETINDEX | ID: biblio-1457923

Resumo

Background: Kernicterus or bilirubin encephalopathy is a condition rarely observed in animal characterized by a yellowish discoloration of the central nervous system. It is a potentially fatal condition due to bilirubin neurotoxic effects caused by the increase of non-conjugated bilirubin pigment, which passes blood brain barrier and has been attributed to an imbalance between albumin and bilirubin levels. Intracellular bilirubin is toxic for cells and can cause decrease in protein synthesis, specially albumin, depression of cell respiration and cellular death. This paper describes kernicterus in a 2-year-old Great Dane female dog.Case: Clinically, the animal showed apathy, lethargy, weight loss and jaundice, which progressed to vomiting and neurological signs characterized by loss of consciousness and eventually coma. Blood parameters were within normal range, except for high levels of alanine aminotransferase (523 U/L), suggesting a liver lesion. The animal was submitted to euthanasia due to the poor prognosis, and at post-mortem examination it showed dehydration and severe jaundice, especially oral, vaginal and ocular mucosas, subcutaneous tissue and blood vessels intima surface. The liver had an accentuated lobular pattern, and the stomach mucosa was reddened. Multiple petechiae were observed in the epicardium, as well as icterus in the blood vessels of the heart. Furthermore, the brain and cerebellum cortex, thalamic region and nuclei region of brainstem showed extensive icteric areas. Microscopically, the liver presented a mononuclear portal hepatitis, centrilobular necrosis and presence of yellowish pigments. The brain had neuronal necrosis, mild vacuolization of the white matter, perineuronal edema and Alzheimer type II astrocytes, while cerebellum showed Purkinje cells necrosis. Hepatic cooper measurement was within range values, and direct imunofluorescence for the detection of Leptospira sp. was negative.[...]


Assuntos
Animais , Cães , Icterícia/veterinária , Kernicterus/patologia , Kernicterus/veterinária , Necrose/veterinária , Neurotoxinas
16.
Acta sci. vet. (Online) ; 46(supl): 1-3, 2018. ilus
Artigo em Inglês | VETINDEX | ID: vti-726510

Resumo

Background: Kernicterus or bilirubin encephalopathy is a condition rarely observed in animal characterized by a yellowish discoloration of the central nervous system. It is a potentially fatal condition due to bilirubin neurotoxic effects caused by the increase of non-conjugated bilirubin pigment, which passes blood brain barrier and has been attributed to an imbalance between albumin and bilirubin levels. Intracellular bilirubin is toxic for cells and can cause decrease in protein synthesis, specially albumin, depression of cell respiration and cellular death. This paper describes kernicterus in a 2-year-old Great Dane female dog.Case: Clinically, the animal showed apathy, lethargy, weight loss and jaundice, which progressed to vomiting and neurological signs characterized by loss of consciousness and eventually coma. Blood parameters were within normal range, except for high levels of alanine aminotransferase (523 U/L), suggesting a liver lesion. The animal was submitted to euthanasia due to the poor prognosis, and at post-mortem examination it showed dehydration and severe jaundice, especially oral, vaginal and ocular mucosas, subcutaneous tissue and blood vessels intima surface. The liver had an accentuated lobular pattern, and the stomach mucosa was reddened. Multiple petechiae were observed in the epicardium, as well as icterus in the blood vessels of the heart. Furthermore, the brain and cerebellum cortex, thalamic region and nuclei region of brainstem showed extensive icteric areas. Microscopically, the liver presented a mononuclear portal hepatitis, centrilobular necrosis and presence of yellowish pigments. The brain had neuronal necrosis, mild vacuolization of the white matter, perineuronal edema and Alzheimer type II astrocytes, while cerebellum showed Purkinje cells necrosis. Hepatic cooper measurement was within range values, and direct imunofluorescence for the detection of Leptospira sp. was negative.[...](AU)


Assuntos
Animais , Cães , Kernicterus/veterinária , Icterícia/veterinária , Necrose/veterinária , Kernicterus/patologia , Neurotoxinas
17.
Vet. Zoot. ; 25: 53-61, 2018.
Artigo em Português | VETINDEX | ID: vti-19828

Resumo

O Botulismo é uma doença causada pela ação da toxina produzida pelo Clostridium botulinumque age no sistema nervoso periférico causando paralisia flácida devido à interrupção da transmissão neuromuscular. Adiciona-se aos sintomas: o decúbito externa ou lateral, visão dupla, turva, além de anorexia, falta de coordenação e ataxia. Em ruminantes, a transmissão dessa doença ocorre principalmente pela ingestão de toxina via alimentos incorretamente armazenados (silagem, ração e feno) e carcaças abandonada nas pastagens. Está diretamente relacionada à osteofagia, decorrente da deficiência mineral (fósforo) nos solos e consequentemente nas forrageiras. O diagnóstico é baseado no histórico e quadro clínico-patológico e o tratamento no fornecimento, o mais breve possível, de solução saturada de hidróxido de magnésio e a administração de soro antibotulínico a fim de facilitar a eliminação da toxina circulante na musculatura do animal. Entretanto, nos quadros mais graves da doença a letalidade em geral pode chegar a 100%. Sendo assim, o objetivo desse trabalho é fornecer informações descritivas sobre as principais causas, sintomas, tratamento e prevenção do Botulismo em búfalos.(AU)


Botulism is a disease caused by the toxin produced by Clostridium botulinumthat acts on the peripheral nervous system causing flaccid paralysis due to the interruption of the neuromuscular transmission. Adds to the symptoms: external or lateral decubitus, doube vision, blurred, besides anorexia, lack of coordination and ataxia. In ruminants, the transmission of this disease occurs mainly by the ingestion of toxin via improperly stored food (silage, feed and hay) and abandoned carcasses in the pastures. It is directly related to osteophage, due to mineral deficiency (phosphorus) in soils and consequently in forages. Thus, the spore develops producing toxins that permeate the porous bones, tendons and ligaments which are later ingested by the animals. The diagnosis is based on the history and clinical-pathological picture and treatment in the shortest possible supply of saturated magnesium hydroxide solution and the administration of antibotulin serum in order to facilitate the elimination of the circulating toxin in the musculature of the animal. However, in the most severe cases of the disease, lethality in general can reach 100%. Thus, the objective of this work is to provide descriptive information on the main causes, symptoms, treatment and prevention of botulism in buffaloes.(AU)


El botulismo es una enfermedad causada por la acción de la toxina producida por el Clostridium botulinum que actúa en el sistema nervioso periférico causando parálisis flácida debido a la interrupción de la transmisión neuromuscular. Se agrega a los síntomas: el decúbito externo o lateral, visión doble, turbia, además de anorexia, falta de coordinación y ataxia. En rumiantes, la transmisión de esta enfermedad ocurre principalmente por la ingestión de toxina vía alimentos incorrectamente almacenados (silaje, ración y heno) y canales abandonadas en los pastizales. Está directamente relacionada a la osteofagia, derivada de la deficiencia mineral (fósforo) en los suelos y consecuentemente en las forrajeras. Así, la espora se desarrolla produciendo toxinas que impregnan en los huesos porosos, tendones y ligamentos que serán posteriormente ingeridos por los animales. El diagnóstico se basa en el historial y cuadro clínico-patológico y el tratamiento en el suministro lo más breve posible de solución saturada de hidróxido de magnesio y la administración de suero antibotulínico para facilitar la eliminación de la toxina circulante en la musculatura del animal. Sin embargo, en los cuadros más graves de la enfermedad la letalidad en general puede llegar al 100%. Por lo tanto, el objetivo de este trabajo es proporcionar información descriptiva sobre las principales causas, síntomas, tratamiento y prevención del botulismo en búfalos.(AU)


Assuntos
Animais , Búfalos , Botulismo/diagnóstico , Botulismo/etiologia , Botulismo/prevenção & controle , Botulismo/veterinária , Infecções por Clostridium/veterinária , Clostridium botulinum , Neurotoxinas
18.
Vet. zootec ; 25: 53-61, 2018.
Artigo em Português | VETINDEX | ID: biblio-1503497

Resumo

O Botulismo é uma doença causada pela ação da toxina produzida pelo Clostridium botulinumque age no sistema nervoso periférico causando paralisia flácida devido à interrupção da transmissão neuromuscular. Adiciona-se aos sintomas: o decúbito externa ou lateral, visão dupla, turva, além de anorexia, falta de coordenação e ataxia. Em ruminantes, a transmissão dessa doença ocorre principalmente pela ingestão de toxina via alimentos incorretamente armazenados (silagem, ração e feno) e carcaças abandonada nas pastagens. Está diretamente relacionada à osteofagia, decorrente da deficiência mineral (fósforo) nos solos e consequentemente nas forrageiras. O diagnóstico é baseado no histórico e quadro clínico-patológico e o tratamento no fornecimento, o mais breve possível, de solução saturada de hidróxido de magnésio e a administração de soro antibotulínico a fim de facilitar a eliminação da toxina circulante na musculatura do animal. Entretanto, nos quadros mais graves da doença a letalidade em geral pode chegar a 100%. Sendo assim, o objetivo desse trabalho é fornecer informações descritivas sobre as principais causas, sintomas, tratamento e prevenção do Botulismo em búfalos.


Botulism is a disease caused by the toxin produced by Clostridium botulinumthat acts on the peripheral nervous system causing flaccid paralysis due to the interruption of the neuromuscular transmission. Adds to the symptoms: external or lateral decubitus, doube vision, blurred, besides anorexia, lack of coordination and ataxia. In ruminants, the transmission of this disease occurs mainly by the ingestion of toxin via improperly stored food (silage, feed and hay) and abandoned carcasses in the pastures. It is directly related to osteophage, due to mineral deficiency (phosphorus) in soils and consequently in forages. Thus, the spore develops producing toxins that permeate the porous bones, tendons and ligaments which are later ingested by the animals. The diagnosis is based on the history and clinical-pathological picture and treatment in the shortest possible supply of saturated magnesium hydroxide solution and the administration of antibotulin serum in order to facilitate the elimination of the circulating toxin in the musculature of the animal. However, in the most severe cases of the disease, lethality in general can reach 100%. Thus, the objective of this work is to provide descriptive information on the main causes, symptoms, treatment and prevention of botulism in buffaloes.


El botulismo es una enfermedad causada por la acción de la toxina producida por el Clostridium botulinum que actúa en el sistema nervioso periférico causando parálisis flácida debido a la interrupción de la transmisión neuromuscular. Se agrega a los síntomas: el decúbito externo o lateral, visión doble, turbia, además de anorexia, falta de coordinación y ataxia. En rumiantes, la transmisión de esta enfermedad ocurre principalmente por la ingestión de toxina vía alimentos incorrectamente almacenados (silaje, ración y heno) y canales abandonadas en los pastizales. Está directamente relacionada a la osteofagia, derivada de la deficiencia mineral (fósforo) en los suelos y consecuentemente en las forrajeras. Así, la espora se desarrolla produciendo toxinas que impregnan en los huesos porosos, tendones y ligamentos que serán posteriormente ingeridos por los animales. El diagnóstico se basa en el historial y cuadro clínico-patológico y el tratamiento en el suministro lo más breve posible de solución saturada de hidróxido de magnesio y la administración de suero antibotulínico para facilitar la eliminación de la toxina circulante en la musculatura del animal. Sin embargo, en los cuadros más graves de la enfermedad la letalidad en general puede llegar al 100%. Por lo tanto, el objetivo de este trabajo es proporcionar información descriptiva sobre las principales causas, síntomas, tratamiento y prevención del botulismo en búfalos.


Assuntos
Animais , Botulismo/diagnóstico , Botulismo/etiologia , Botulismo/prevenção & controle , Botulismo/veterinária , Búfalos , Clostridium botulinum , Infecções por Clostridium/veterinária , Neurotoxinas
19.
Pesqui. vet. bras ; 37(12): 1369-1372, dez. 2017. ilus
Artigo em Português | LILACS, VETINDEX | ID: biblio-895402

Resumo

O botulismo nos animais domésticos é uma intoxicação fatal ocasionada pela ingestão da neurotoxina de Clostridium botulinum. Este trabalho descreve os aspectos epidemiológicos, clínicos e laboratoriais de um surto de botulismo tipo C em equinos de uma propriedade rural no município de Porto Alegre, Rio Grande do Sul. Do total de 20 equinos, sete (35%) foram acometidos e morreram, e um eutanasiado após apresentar prognóstico desfavorável. Os cavalos, criados extensivamente, apresentaram sinais clínicos semelhantes, caracterizados por dificuldade de locomoção, letargia, paralisia flácida progressiva, tremores musculares, sudorese, ataxia e quedas frequentes. Quatro equinos foram necropsiados e não apresentaram lesões macroscópicas e microscópicas significativas. Amostras de encéfalo e medula espinhal foram enviados para realização do exame de imunofluorescência direta e foram negativas para o vírus da raiva. Fragmentos de fígado dos quatro equinos necropsiados, amostras de sedimento do solo e água suspeita de dois açudes da propriedade foram avaliados pelo exame direto para a verificação da presença de toxina botulínica e/ou esporos de Clostridium botulinum. No bioensaio em camundongos e soroneutralização pode-se detectar toxina botulínica tipo C em uma amostra de fígado, ao passo que as amostras de sedimento do solo e água foram negativas em todos os ensaios. Com base nos dados epidemiológicos, clínicos e laboratoriais pode-se estabelecer o diagnóstico etiológico de botulismo causado pela toxina botulínica do tipo C, e sugere-se, a água estagnada dos açudes como aprovável fonte da toxina.(AU)


Botulism in domestic animals is a fatal poisoning caused by ingestion of Clostridium botulinum neurotoxin. This paper describes epidemiological, clinical and laboratorial aspects of a botulism type C outbreak in horses in a farm in Porto Alegre, Rio Grande do Sul. From a total of 20 horses, seven (35%) were affected and died, and one was euthanized after presenting poor prognosis. The horses, which were raised extensively, showed similar clinical signs, that included limited mobility, lethargy, progressive flaccid paralysis, muscle tremors, sweating, ataxia, and frequent falls. Four horses were necropsied and did not show significant macroscopic and microscopic lesions. Brain and spinal cord samples were analyzed by direct immunofluorescence and were negative for rabies. Liver fragments from the four necropsied horses, soil sediment samples, and suspect water from two reservoirs were evaluated by direct exam to verify the botulinum toxin and/or Clostridium botulinum spores presence. In the serum neutralization bioassay in mice, the type C botulinum toxin was detected in one liver sample, at the same time that the soil sediment and water samples were negative in all assays. Based on the epidemiological, clinical and laboratory data the etiological diagnosis of botulism was established by being caused by botulinum toxin type C, and it is suggested that the stagnant water of the dams is the likely source of the toxin.(AU)


Assuntos
Animais , Botulismo/epidemiologia , Surtos de Doenças/veterinária , Clostridium botulinum tipo C , Cavalos/microbiologia , Brasil/epidemiologia , Neurotoxinas
20.
Pesqui. vet. bras ; 37(12): 1369-1372, dez. 2017. ilus
Artigo em Português | VETINDEX | ID: vti-743391

Resumo

O botulismo nos animais domésticos é uma intoxicação fatal ocasionada pela ingestão da neurotoxina de Clostridium botulinum. Este trabalho descreve os aspectos epidemiológicos, clínicos e laboratoriais de um surto de botulismo tipo C em equinos de uma propriedade rural no município de Porto Alegre, Rio Grande do Sul. Do total de 20 equinos, sete (35%) foram acometidos e morreram, e um eutanasiado após apresentar prognóstico desfavorável. Os cavalos, criados extensivamente, apresentaram sinais clínicos semelhantes, caracterizados por dificuldade de locomoção, letargia, paralisia flácida progressiva, tremores musculares, sudorese, ataxia e quedas frequentes. Quatro equinos foram necropsiados e não apresentaram lesões macroscópicas e microscópicas significativas. Amostras de encéfalo e medula espinhal foram enviados para realização do exame de imunofluorescência direta e foram negativas para o vírus da raiva. Fragmentos de fígado dos quatro equinos necropsiados, amostras de sedimento do solo e água suspeita de dois açudes da propriedade foram avaliados pelo exame direto para a verificação da presença de toxina botulínica e/ou esporos de Clostridium botulinum. No bioensaio em camundongos e soroneutralização pode-se detectar toxina botulínica tipo C em uma amostra de fígado, ao passo que as amostras de sedimento do solo e água foram negativas em todos os ensaios. Com base nos dados epidemiológicos, clínicos e laboratoriais pode-se estabelecer o diagnóstico etiológico de botulismo causado pela toxina botulínica do tipo C, e sugere-se, a água estagnada dos açudes como aprovável fonte da toxina.(AU)


Botulism in domestic animals is a fatal poisoning caused by ingestion of Clostridium botulinum neurotoxin. This paper describes epidemiological, clinical and laboratorial aspects of a botulism type C outbreak in horses in a farm in Porto Alegre, Rio Grande do Sul. From a total of 20 horses, seven (35%) were affected and died, and one was euthanized after presenting poor prognosis. The horses, which were raised extensively, showed similar clinical signs, that included limited mobility, lethargy, progressive flaccid paralysis, muscle tremors, sweating, ataxia, and frequent falls. Four horses were necropsied and did not show significant macroscopic and microscopic lesions. Brain and spinal cord samples were analyzed by direct immunofluorescence and were negative for rabies. Liver fragments from the four necropsied horses, soil sediment samples, and suspect water from two reservoirs were evaluated by direct exam to verify the botulinum toxin and/or Clostridium botulinum spores presence. In the serum neutralization bioassay in mice, the type C botulinum toxin was detected in one liver sample, at the same time that the soil sediment and water samples were negative in all assays. Based on the epidemiological, clinical and laboratory data the etiological diagnosis of botulism was established by being caused by botulinum toxin type C, and it is suggested that the stagnant water of the dams is the likely source of the toxin.(AU)


Assuntos
Animais , Botulismo/epidemiologia , Surtos de Doenças/veterinária , Clostridium botulinum tipo C , Cavalos/microbiologia , Brasil/epidemiologia , Neurotoxinas
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