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African adders: partial characterization of snake venoms from three Bitis species of medical importance and their neutralization by experimental equine antivenoms.

Paixão-Cavalcante D, Kuniyoshi AK, Portaro FC, da Silva WD, Tambourgi DV - PLoS Negl Trop Dis (2015)

Bottom Line: The accidents are severe and the victims often have a poor prognosis due to the lack of effective specific therapies.Experimental antivenoms produced against B. arietans venom or Bitis g. rhinoceros plus B. nasicornis venoms cross-reacted with the venoms from the three species and blocked, in different degrees, all the enzymatic activities in which they were tested.We also demonstrated that horse antivenoms produced against B. arietans or B. g. rhinoceros plus B. nasicornis venoms can blocked some of the toxic activities of these venoms.

View Article: PubMed Central - PubMed

Affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, São Paulo, Brazil.

ABSTRACT

Background: An alarming number of fatal accidents involving snakes are annually reported in Africa and most of the victims suffer from permanent local tissue damage and chronic disabilities. Envenomation by snakes belonging to the genus Bitis, Viperidae family, are common in Sub-Saharan Africa. The accidents are severe and the victims often have a poor prognosis due to the lack of effective specific therapies. In this study we have biochemically characterized venoms from three different species of Bitis, i.e., Bitis arietans, Bitis gabonica rhinoceros and Bitis nasicornis, involved in the majority of the human accidents in Africa, and analyzed the in vitro neutralizing ability of two experimental antivenoms.

Methodology/principal findings: The data indicate that all venoms presented phospholipase, hyaluronidase and fibrinogenolytic activities and cleaved efficiently the FRET substrate Abz-RPPGFSPFRQ-EDDnp and angiotensin I, generating angiotensin 1-7. Gelatinolytic activity was only observed in the venoms of B. arietans and B. nasicornis. The treatment of the venoms with protease inhibitors indicated that Bitis venoms possess metallo and serinoproteases enzymes, which may be involved in the different biological activities here evaluated. Experimental antivenoms produced against B. arietans venom or Bitis g. rhinoceros plus B. nasicornis venoms cross-reacted with the venoms from the three species and blocked, in different degrees, all the enzymatic activities in which they were tested.

Conclusion: These results suggest that the venoms of the three Bitis species, involved in accidents with humans in the Sub-Saharan Africa, contain a mixture of various enzymes that may act in the generation and development of some of the clinical manifestations of the envenomations. We also demonstrated that horse antivenoms produced against B. arietans or B. g. rhinoceros plus B. nasicornis venoms can blocked some of the toxic activities of these venoms.

No MeSH data available.


Related in: MedlinePlus

Titers and cross-reactivity of antivenoms raised against B. arietans and Bitis g. rhinoceros plus B. nasicornis venoms.[A]ELISA: ELISA plates were coated with 1.0 μg of the Bitis spp venoms/well and incubated with different dilutions of the horse experimental antivenoms produced against B. arietans venom (α-Ba antivenom), B. g. rhinoceros plus B. nasicornis venoms (α-Br + Bn antivenom) or with the botulinic toxin antiserum (control), followed by anti-horse IgG-HRPO-conjugated. The results were expressed as the mean of absorbance value ± SD. [B]Western Blot: venom samples (5 μg) from B. arietans (Ba), B. g. rhinoceros (Br) and B. nasicornis (Bn) snakes were separated by SDS-PAGE, electrotransfered to nitrocellulose membranes and incubated with the antivenoms raised against B. arietans (α-Ba), B. g. rhinoceros plus B. nasicornis (α-Br+Bn) or with botulinic toxin antiserum (control) diluted 1:2000 followed by GAH/Ig-AP. The reactions were revealed with NBT and BCIP.
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pntd.0003419.g006: Titers and cross-reactivity of antivenoms raised against B. arietans and Bitis g. rhinoceros plus B. nasicornis venoms.[A]ELISA: ELISA plates were coated with 1.0 μg of the Bitis spp venoms/well and incubated with different dilutions of the horse experimental antivenoms produced against B. arietans venom (α-Ba antivenom), B. g. rhinoceros plus B. nasicornis venoms (α-Br + Bn antivenom) or with the botulinic toxin antiserum (control), followed by anti-horse IgG-HRPO-conjugated. The results were expressed as the mean of absorbance value ± SD. [B]Western Blot: venom samples (5 μg) from B. arietans (Ba), B. g. rhinoceros (Br) and B. nasicornis (Bn) snakes were separated by SDS-PAGE, electrotransfered to nitrocellulose membranes and incubated with the antivenoms raised against B. arietans (α-Ba), B. g. rhinoceros plus B. nasicornis (α-Br+Bn) or with botulinic toxin antiserum (control) diluted 1:2000 followed by GAH/Ig-AP. The reactions were revealed with NBT and BCIP.

Mentions: α-Ba and α-Br + Bn antivenoms presented high specific antibody titers and cross-reactivity with others Bitis ssp venoms (Fig. 6A). Western Blot analysis showed that α-Ba and α-Br + Bn antivenoms efficiently immune reacted to a vast number of proteins in the crude venoms (Fig. 6B). Surprisingly, the western blot analysis showed that α-Ba antivenom better recognized the bands with low molecular weight in the venoms of B. g. rhinoceros and B. nasicornis than the α-Br + Bn antivenom (Fig. 6B).


African adders: partial characterization of snake venoms from three Bitis species of medical importance and their neutralization by experimental equine antivenoms.

Paixão-Cavalcante D, Kuniyoshi AK, Portaro FC, da Silva WD, Tambourgi DV - PLoS Negl Trop Dis (2015)

Titers and cross-reactivity of antivenoms raised against B. arietans and Bitis g. rhinoceros plus B. nasicornis venoms.[A]ELISA: ELISA plates were coated with 1.0 μg of the Bitis spp venoms/well and incubated with different dilutions of the horse experimental antivenoms produced against B. arietans venom (α-Ba antivenom), B. g. rhinoceros plus B. nasicornis venoms (α-Br + Bn antivenom) or with the botulinic toxin antiserum (control), followed by anti-horse IgG-HRPO-conjugated. The results were expressed as the mean of absorbance value ± SD. [B]Western Blot: venom samples (5 μg) from B. arietans (Ba), B. g. rhinoceros (Br) and B. nasicornis (Bn) snakes were separated by SDS-PAGE, electrotransfered to nitrocellulose membranes and incubated with the antivenoms raised against B. arietans (α-Ba), B. g. rhinoceros plus B. nasicornis (α-Br+Bn) or with botulinic toxin antiserum (control) diluted 1:2000 followed by GAH/Ig-AP. The reactions were revealed with NBT and BCIP.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340965&req=5

pntd.0003419.g006: Titers and cross-reactivity of antivenoms raised against B. arietans and Bitis g. rhinoceros plus B. nasicornis venoms.[A]ELISA: ELISA plates were coated with 1.0 μg of the Bitis spp venoms/well and incubated with different dilutions of the horse experimental antivenoms produced against B. arietans venom (α-Ba antivenom), B. g. rhinoceros plus B. nasicornis venoms (α-Br + Bn antivenom) or with the botulinic toxin antiserum (control), followed by anti-horse IgG-HRPO-conjugated. The results were expressed as the mean of absorbance value ± SD. [B]Western Blot: venom samples (5 μg) from B. arietans (Ba), B. g. rhinoceros (Br) and B. nasicornis (Bn) snakes were separated by SDS-PAGE, electrotransfered to nitrocellulose membranes and incubated with the antivenoms raised against B. arietans (α-Ba), B. g. rhinoceros plus B. nasicornis (α-Br+Bn) or with botulinic toxin antiserum (control) diluted 1:2000 followed by GAH/Ig-AP. The reactions were revealed with NBT and BCIP.
Mentions: α-Ba and α-Br + Bn antivenoms presented high specific antibody titers and cross-reactivity with others Bitis ssp venoms (Fig. 6A). Western Blot analysis showed that α-Ba and α-Br + Bn antivenoms efficiently immune reacted to a vast number of proteins in the crude venoms (Fig. 6B). Surprisingly, the western blot analysis showed that α-Ba antivenom better recognized the bands with low molecular weight in the venoms of B. g. rhinoceros and B. nasicornis than the α-Br + Bn antivenom (Fig. 6B).

Bottom Line: The accidents are severe and the victims often have a poor prognosis due to the lack of effective specific therapies.Experimental antivenoms produced against B. arietans venom or Bitis g. rhinoceros plus B. nasicornis venoms cross-reacted with the venoms from the three species and blocked, in different degrees, all the enzymatic activities in which they were tested.We also demonstrated that horse antivenoms produced against B. arietans or B. g. rhinoceros plus B. nasicornis venoms can blocked some of the toxic activities of these venoms.

View Article: PubMed Central - PubMed

Affiliation: Immunochemistry Laboratory, Butantan Institute, São Paulo, São Paulo, Brazil.

ABSTRACT

Background: An alarming number of fatal accidents involving snakes are annually reported in Africa and most of the victims suffer from permanent local tissue damage and chronic disabilities. Envenomation by snakes belonging to the genus Bitis, Viperidae family, are common in Sub-Saharan Africa. The accidents are severe and the victims often have a poor prognosis due to the lack of effective specific therapies. In this study we have biochemically characterized venoms from three different species of Bitis, i.e., Bitis arietans, Bitis gabonica rhinoceros and Bitis nasicornis, involved in the majority of the human accidents in Africa, and analyzed the in vitro neutralizing ability of two experimental antivenoms.

Methodology/principal findings: The data indicate that all venoms presented phospholipase, hyaluronidase and fibrinogenolytic activities and cleaved efficiently the FRET substrate Abz-RPPGFSPFRQ-EDDnp and angiotensin I, generating angiotensin 1-7. Gelatinolytic activity was only observed in the venoms of B. arietans and B. nasicornis. The treatment of the venoms with protease inhibitors indicated that Bitis venoms possess metallo and serinoproteases enzymes, which may be involved in the different biological activities here evaluated. Experimental antivenoms produced against B. arietans venom or Bitis g. rhinoceros plus B. nasicornis venoms cross-reacted with the venoms from the three species and blocked, in different degrees, all the enzymatic activities in which they were tested.

Conclusion: These results suggest that the venoms of the three Bitis species, involved in accidents with humans in the Sub-Saharan Africa, contain a mixture of various enzymes that may act in the generation and development of some of the clinical manifestations of the envenomations. We also demonstrated that horse antivenoms produced against B. arietans or B. g. rhinoceros plus B. nasicornis venoms can blocked some of the toxic activities of these venoms.

No MeSH data available.


Related in: MedlinePlus