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Juan J. Calvete - One of the best experts on this subject based on the ideXlab platform.
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antivenomic assessment of the immunological reactivity of echitab plus icp an antivenom for the treatment of snakebite envenoming in sub saharan africa
American Journal of Tropical Medicine and Hygiene, 2010Co-Authors: Juan J. Calvete, Robert A. Harrison, Libia Sanz, Alvaro Segura, Mauren Villalta, Maria Herrera, Guillermo Leon, Nandul Durfa, Abdusalami Nasidi, David R G TheakstonAbstract:The immunoreactivity of EchiTAb-Plus-ICP, an antivenom developed for the treatment of snakebite envenoming in sub-Saharan Africa, to venoms of seven Echis and Bitis species, was assessed by "antivenomics." This proteomic approach is based on the ability of an antivenom to immunodeplete homologous or heterologous venom proteins. Our results show an extensive cross-reactivity of this antivenom against all Echis and Bitis venoms studied, as revealed by the complete immunodepletion of the majority of venom components, including metalloproteinases, serine proteinases, C-type lectin-like proteins, some phospholipases A(2) and L-amino acid oxidase. However, some phospholipases A(2), disintegrins and proteinase inhibitors were immunodepleted to only a partial extent. These results support the hypothesis that immunizing horses with a mixture of the venoms of Echis ocellatus, Bitis arietans, and Naja nigricollis generates antibodies capable of recognizing the majority of components of medically-relevant homologous and heterologous viperid venoms of the genera Bitis and Echis from sub-Saharan Africa.
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preclinical assessment of the efficacy of a new antivenom echitab plus icp for the treatment of viper envenoming in sub saharan africa
Toxicon, 2010Co-Authors: Alvaro Segura, Robert A. Harrison, Juan J. Calvete, Mauren Villalta, Maria Herrera, Guillermo Leon, Nandul Durfa, Abdusalami Nasidi, David R G Theakston, David A. WarrellAbstract:A preclinical assessment was performed on the neutralizing efficacy of a whole IgG polyspecific antivenom (EchiTAb-Plus-ICP®), designed for the treatment of snakebite envenomings in Nigeria. It was generated by immunizing horses with the venoms of Echis ocellatus, Bitis arietans and Naja nigricollis, the most medically important species in Nigeria. Antivenom was tested against the venoms of E. ocellatus, Echis leucogaster, Echis pyramidum leakeyi, B. arietans, Bitis gabonica, Bitis rhinoceros and Bitis nasicornis. The neutralization of the venom toxins responsible for the lethal, hemorrhagic, coagulant and local necrotizing activities was assessed, since these are the most significant effects that characterize envenoming by these species. Echis sp venoms exerted lethal, hemorrhagic, coagulant and necrotizing effects, whereas the Bitis sp venoms tested induced lethality, hemorrhage and necrosis, but were devoid of coagulant activity. The antivenom was effective in the neutralization of all effects tested in all venoms. Highest neutralization was achieved against the venoms of E. ocellatus and B. arietans, and the lowest neutralizing potency was against the venom of B. nasicornis, a species that has a low clinical relevance. It is concluded that EchiTAb-Plus-ICP®, whilst specifically designed for Nigeria, has a good preclinical neutralizing profile against homologous and heterologous viperid venoms from other sub-Saharan African locations. It therefore constitutes a promising therapeutic option for the treatment of snakebite envenoming in this region.
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Snake venomics of Bitis species reveals large intragenus venom toxin composition variation: application to taxonomy of congeneric taxa.
Journal of proteome research, 2007Co-Authors: Juan J. Calvete, José Escolano, Libia SanzAbstract:The protein composition of the venoms of the West African Gaboon viper (Bitis gabonica rhinoceros), the rhinoceros viper (Bitis nasicornis), and the horned puff adder (Bitis caudalis) were analyzed...
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Snake venomics of Bitis gabonica gabonica. Protein family composition, subunit organization of venom toxins, and characterization of dimeric disintegrins Bitisgabonin-1 and Bitisgabonin-2.
Journal of proteome research, 2007Co-Authors: Juan J. Calvete, Cezary Marcinkiewicz, Libia SanzAbstract:The protein composition of the venom of the East African Gaboon viper (Bitis gabonica gabonica) was analyzed using RP-HPLC, N-terminal sequencing, MALDI-TOF peptide mass fingerprinting, and CID−MS/...
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Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin
Journal of Molecular Evolution, 2006Co-Authors: Paula Juárez, Simon C. Wagstaff, Robert A. Harrison, Jenny Oliver, Libia Sanz, Juan J. CalveteAbstract:We report the cloning and sequence analysis of BA-5A from a venom gland cDNA library of the puff adder, Bitis arietans , that encodes a novel ECD-disintegrin-like domain. BA-5A is a unique PII disintegrin. It contains the 16 cysteine residues that are conserved in all known disintegrin-like domains of ADAM proteins and snake venom metalloproteinases but lacks the cysteine-rich domain. These features suggest that BA-5A may represent an intermediate in the evolutionary pathway of the long disintegrin Bitistatin and that removal of the cysteine-rich domain and loss of the PIII-specific disulfide bond were separate events along the structural diversification pathway of disintegrins, the former predating the latter. The protein family composition of the Bitis arietans venom, as determined by combination of reversed-phase HPLC and proteomic analysis, was as follows: Zn^2+-metalloproteinase (38.5%), serine proteinase (19.5%), disintegrin (17.8%), C-type lectin-like (13.2%), PLA_2 (4.3%), Kunitz-type inhibitor (4.1%), cystatin (1.7%), and unknown (0.9%). BA-5A could not be detected in the venom proteome of Bitis arietans . The occurrence of this very low-abundance (< 0.05%) or nonexpressed disintegrin transcript indicates a hitherto unrecognized structural diversity of this protein family. Whether BA-5A plays a physiological role or represents an orphan protein which could eventually evolve a role in the adaptation of snakes to changing ecological niches and prey habits deserves further investigation.
Libia Sanz - One of the best experts on this subject based on the ideXlab platform.
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antivenomic assessment of the immunological reactivity of echitab plus icp an antivenom for the treatment of snakebite envenoming in sub saharan africa
American Journal of Tropical Medicine and Hygiene, 2010Co-Authors: Juan J. Calvete, Robert A. Harrison, Libia Sanz, Alvaro Segura, Mauren Villalta, Maria Herrera, Guillermo Leon, Nandul Durfa, Abdusalami Nasidi, David R G TheakstonAbstract:The immunoreactivity of EchiTAb-Plus-ICP, an antivenom developed for the treatment of snakebite envenoming in sub-Saharan Africa, to venoms of seven Echis and Bitis species, was assessed by "antivenomics." This proteomic approach is based on the ability of an antivenom to immunodeplete homologous or heterologous venom proteins. Our results show an extensive cross-reactivity of this antivenom against all Echis and Bitis venoms studied, as revealed by the complete immunodepletion of the majority of venom components, including metalloproteinases, serine proteinases, C-type lectin-like proteins, some phospholipases A(2) and L-amino acid oxidase. However, some phospholipases A(2), disintegrins and proteinase inhibitors were immunodepleted to only a partial extent. These results support the hypothesis that immunizing horses with a mixture of the venoms of Echis ocellatus, Bitis arietans, and Naja nigricollis generates antibodies capable of recognizing the majority of components of medically-relevant homologous and heterologous viperid venoms of the genera Bitis and Echis from sub-Saharan Africa.
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Snake venomics of Bitis species reveals large intragenus venom toxin composition variation: application to taxonomy of congeneric taxa.
Journal of proteome research, 2007Co-Authors: Juan J. Calvete, José Escolano, Libia SanzAbstract:The protein composition of the venoms of the West African Gaboon viper (Bitis gabonica rhinoceros), the rhinoceros viper (Bitis nasicornis), and the horned puff adder (Bitis caudalis) were analyzed...
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Snake venomics of Bitis gabonica gabonica. Protein family composition, subunit organization of venom toxins, and characterization of dimeric disintegrins Bitisgabonin-1 and Bitisgabonin-2.
Journal of proteome research, 2007Co-Authors: Juan J. Calvete, Cezary Marcinkiewicz, Libia SanzAbstract:The protein composition of the venom of the East African Gaboon viper (Bitis gabonica gabonica) was analyzed using RP-HPLC, N-terminal sequencing, MALDI-TOF peptide mass fingerprinting, and CID−MS/...
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Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin
Journal of Molecular Evolution, 2006Co-Authors: Paula Juárez, Simon C. Wagstaff, Robert A. Harrison, Jenny Oliver, Libia Sanz, Juan J. CalveteAbstract:We report the cloning and sequence analysis of BA-5A from a venom gland cDNA library of the puff adder, Bitis arietans , that encodes a novel ECD-disintegrin-like domain. BA-5A is a unique PII disintegrin. It contains the 16 cysteine residues that are conserved in all known disintegrin-like domains of ADAM proteins and snake venom metalloproteinases but lacks the cysteine-rich domain. These features suggest that BA-5A may represent an intermediate in the evolutionary pathway of the long disintegrin Bitistatin and that removal of the cysteine-rich domain and loss of the PIII-specific disulfide bond were separate events along the structural diversification pathway of disintegrins, the former predating the latter. The protein family composition of the Bitis arietans venom, as determined by combination of reversed-phase HPLC and proteomic analysis, was as follows: Zn^2+-metalloproteinase (38.5%), serine proteinase (19.5%), disintegrin (17.8%), C-type lectin-like (13.2%), PLA_2 (4.3%), Kunitz-type inhibitor (4.1%), cystatin (1.7%), and unknown (0.9%). BA-5A could not be detected in the venom proteome of Bitis arietans . The occurrence of this very low-abundance (< 0.05%) or nonexpressed disintegrin transcript indicates a hitherto unrecognized structural diversity of this protein family. Whether BA-5A plays a physiological role or represents an orphan protein which could eventually evolve a role in the adaptation of snakes to changing ecological niches and prey habits deserves further investigation.
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Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin
Journal of Molecular Evolution, 2006Co-Authors: Paula Juárez, Simon C. Wagstaff, Robert A. Harrison, Jenny Oliver, Libia Sanz, Juan J. CalveteAbstract:We report the cloning and sequence analysis of BA-5A from a venom gland cDNA library of the puff adder, Bitis arietans , that encodes a novel ECD-disintegrin-like domain. BA-5A is a unique PII disintegrin. It contains the 16 cysteine residues that are conserved in all known disintegrin-like domains of ADAM proteins and snake venom metalloproteinases but lacks the cysteine-rich domain. These features suggest that BA-5A may represent an intermediate in the evolutionary pathway of the long disintegrin Bitistatin and that removal of the cysteine-rich domain and loss of the PIII-specific disulfide bond were separate events along the structural diversification pathway of disintegrins, the former predating the latter. The protein family composition of the Bitis arietans venom, as determined by combination of reversed-phase HPLC and proteomic analysis, was as follows: Zn^2+-metalloproteinase (38.5%), serine proteinase (19.5%), disintegrin (17.8%), C-type lectin-like (13.2%), PLA_2 (4.3%), Kunitz-type inhibitor (4.1%), cystatin (1.7%), and unknown (0.9%). BA-5A could not be detected in the venom proteome of Bitis arietans . The occurrence of this very low-abundance (< 0.05%) or nonexpressed disintegrin transcript indicates a hitherto unrecognized structural diversity of this protein family. Whether BA-5A plays a physiological role or represents an orphan protein which could eventually evolve a role in the adaptation of snakes to changing ecological niches and prey habits deserves further investigation.
Ronald L. Gutberlet - One of the best experts on this subject based on the ideXlab platform.
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Recovering the evolutionary history of Africa’s most diverse viper genus: morphological and molecular phylogeny of Bitis (Reptilia: Squamata: Viperidae)
Organisms Diversity & Evolution, 2015Co-Authors: Rod D. Wittenberg, Robert C. Jadin, Allyson M. Fenwick, Ronald L. GutberletAbstract:Assessing evolutionary relationships among wide-ranging species can be particularly beneficial to our understanding of speciation patterns and biogeography of taxa, with broad implications for conservation and applications for human health. Integrative phylogenetic analyses that incorporate multiple independent datasets (e.g., DNA, protein, phenotype) can resolve many problematic issues in systematics such as cryptic diversity and incongruence between datasets. Vipers in the genus Bitis are widely distributed throughout much of sub-Saharan Africa, filling a variety of ecological niches and presenting an important public health problem. However, evolutionary relationships among this medically and ecologically important genus have not been fully resolved due to inadequate taxon sampling and lack of informative characters. Here, we conduct the first phylogenetic study incorporating complete sampling of known species within the genus Bitis . Using morphological, molecular, and combined approaches under multiple criteria, we recovered many of the species groups detected by previous investigators, further validating four currently recognized subgenera. Bitis arietans and Bitis worthingtoni appear to be early-diverging, monotypic lineages, while the “big Bitis ” group and the small southern African species form distinct clades. Although our study provides additional information regarding the interspecific relationships within Bitis , the placement of Bitis albanica , Bitis heraldica , and Bitis inornata remains problematic. This study enhances our understanding of the evolutionary history of species within the genus Bitis incorporating a combined evidence approach to phylogenetics.
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recovering the evolutionary history of africa s most diverse viper genus morphological and molecular phylogeny of Bitis reptilia squamata viperidae
Organisms Diversity & Evolution, 2015Co-Authors: Rod D. Wittenberg, Robert C. Jadin, Allyson M. Fenwick, Ronald L. GutberletAbstract:Assessing evolutionary relationships among wide-ranging species can be particularly beneficial to our understanding of speciation patterns and biogeography of taxa, with broad implications for conservation and applications for human health. Integrative phylogenetic analyses that incorporate multiple independent datasets (e.g., DNA, protein, phenotype) can resolve many problematic issues in systematics such as cryptic diversity and incongruence between datasets. Vipers in the genus Bitis are widely distributed throughout much of sub-Saharan Africa, filling a variety of ecological niches and presenting an important public health problem. However, evolutionary relationships among this medically and ecologically important genus have not been fully resolved due to inadequate taxon sampling and lack of informative characters. Here, we conduct the first phylogenetic study incorporating complete sampling of known species within the genus Bitis. Using morphological, molecular, and combined approaches under multiple criteria, we recovered many of the species groups detected by previous investigators, further validating four currently recognized subgenera. Bitis arietans and Bitis worthingtoni appear to be early-diverging, monotypic lineages, while the “big Bitis” group and the small southern African species form distinct clades. Although our study provides additional information regarding the interspecific relationships within Bitis, the placement of Bitis albanica, Bitis heraldica, and Bitis inornata remains problematic. This study enhances our understanding of the evolutionary history of species within the genus Bitis incorporating a combined evidence approach to phylogenetics.
Robert A. Harrison - One of the best experts on this subject based on the ideXlab platform.
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Bitis arietans primers.
2016Co-Authors: Gareth Whiteley, Rhiannon A. E. Logan, Kam-yin D. Leung, Fiona J. Newberry, Paul D. Rowley, John P. Dunbar, Simon C. Wagstaff, Nicholas R. Casewell, Robert A. HarrisonAbstract:Bitis arietans primers.
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antivenomic assessment of the immunological reactivity of echitab plus icp an antivenom for the treatment of snakebite envenoming in sub saharan africa
American Journal of Tropical Medicine and Hygiene, 2010Co-Authors: Juan J. Calvete, Robert A. Harrison, Libia Sanz, Alvaro Segura, Mauren Villalta, Maria Herrera, Guillermo Leon, Nandul Durfa, Abdusalami Nasidi, David R G TheakstonAbstract:The immunoreactivity of EchiTAb-Plus-ICP, an antivenom developed for the treatment of snakebite envenoming in sub-Saharan Africa, to venoms of seven Echis and Bitis species, was assessed by "antivenomics." This proteomic approach is based on the ability of an antivenom to immunodeplete homologous or heterologous venom proteins. Our results show an extensive cross-reactivity of this antivenom against all Echis and Bitis venoms studied, as revealed by the complete immunodepletion of the majority of venom components, including metalloproteinases, serine proteinases, C-type lectin-like proteins, some phospholipases A(2) and L-amino acid oxidase. However, some phospholipases A(2), disintegrins and proteinase inhibitors were immunodepleted to only a partial extent. These results support the hypothesis that immunizing horses with a mixture of the venoms of Echis ocellatus, Bitis arietans, and Naja nigricollis generates antibodies capable of recognizing the majority of components of medically-relevant homologous and heterologous viperid venoms of the genera Bitis and Echis from sub-Saharan Africa.
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preclinical assessment of the efficacy of a new antivenom echitab plus icp for the treatment of viper envenoming in sub saharan africa
Toxicon, 2010Co-Authors: Alvaro Segura, Robert A. Harrison, Juan J. Calvete, Mauren Villalta, Maria Herrera, Guillermo Leon, Nandul Durfa, Abdusalami Nasidi, David R G Theakston, David A. WarrellAbstract:A preclinical assessment was performed on the neutralizing efficacy of a whole IgG polyspecific antivenom (EchiTAb-Plus-ICP®), designed for the treatment of snakebite envenomings in Nigeria. It was generated by immunizing horses with the venoms of Echis ocellatus, Bitis arietans and Naja nigricollis, the most medically important species in Nigeria. Antivenom was tested against the venoms of E. ocellatus, Echis leucogaster, Echis pyramidum leakeyi, B. arietans, Bitis gabonica, Bitis rhinoceros and Bitis nasicornis. The neutralization of the venom toxins responsible for the lethal, hemorrhagic, coagulant and local necrotizing activities was assessed, since these are the most significant effects that characterize envenoming by these species. Echis sp venoms exerted lethal, hemorrhagic, coagulant and necrotizing effects, whereas the Bitis sp venoms tested induced lethality, hemorrhage and necrosis, but were devoid of coagulant activity. The antivenom was effective in the neutralization of all effects tested in all venoms. Highest neutralization was achieved against the venoms of E. ocellatus and B. arietans, and the lowest neutralizing potency was against the venom of B. nasicornis, a species that has a low clinical relevance. It is concluded that EchiTAb-Plus-ICP®, whilst specifically designed for Nigeria, has a good preclinical neutralizing profile against homologous and heterologous viperid venoms from other sub-Saharan African locations. It therefore constitutes a promising therapeutic option for the treatment of snakebite envenoming in this region.
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Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin
Journal of Molecular Evolution, 2006Co-Authors: Paula Juárez, Simon C. Wagstaff, Robert A. Harrison, Jenny Oliver, Libia Sanz, Juan J. CalveteAbstract:We report the cloning and sequence analysis of BA-5A from a venom gland cDNA library of the puff adder, Bitis arietans , that encodes a novel ECD-disintegrin-like domain. BA-5A is a unique PII disintegrin. It contains the 16 cysteine residues that are conserved in all known disintegrin-like domains of ADAM proteins and snake venom metalloproteinases but lacks the cysteine-rich domain. These features suggest that BA-5A may represent an intermediate in the evolutionary pathway of the long disintegrin Bitistatin and that removal of the cysteine-rich domain and loss of the PIII-specific disulfide bond were separate events along the structural diversification pathway of disintegrins, the former predating the latter. The protein family composition of the Bitis arietans venom, as determined by combination of reversed-phase HPLC and proteomic analysis, was as follows: Zn^2+-metalloproteinase (38.5%), serine proteinase (19.5%), disintegrin (17.8%), C-type lectin-like (13.2%), PLA_2 (4.3%), Kunitz-type inhibitor (4.1%), cystatin (1.7%), and unknown (0.9%). BA-5A could not be detected in the venom proteome of Bitis arietans . The occurrence of this very low-abundance (< 0.05%) or nonexpressed disintegrin transcript indicates a hitherto unrecognized structural diversity of this protein family. Whether BA-5A plays a physiological role or represents an orphan protein which could eventually evolve a role in the adaptation of snakes to changing ecological niches and prey habits deserves further investigation.
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Molecular Cloning of Disintegrin-like Transcript BA-5A from a Bitis arietans Venom Gland cDNA Library: A Putative Intermediate in the Evolution of the Long-Chain Disintegrin Bitistatin
Journal of Molecular Evolution, 2006Co-Authors: Paula Juárez, Simon C. Wagstaff, Robert A. Harrison, Jenny Oliver, Libia Sanz, Juan J. CalveteAbstract:We report the cloning and sequence analysis of BA-5A from a venom gland cDNA library of the puff adder, Bitis arietans , that encodes a novel ECD-disintegrin-like domain. BA-5A is a unique PII disintegrin. It contains the 16 cysteine residues that are conserved in all known disintegrin-like domains of ADAM proteins and snake venom metalloproteinases but lacks the cysteine-rich domain. These features suggest that BA-5A may represent an intermediate in the evolutionary pathway of the long disintegrin Bitistatin and that removal of the cysteine-rich domain and loss of the PIII-specific disulfide bond were separate events along the structural diversification pathway of disintegrins, the former predating the latter. The protein family composition of the Bitis arietans venom, as determined by combination of reversed-phase HPLC and proteomic analysis, was as follows: Zn^2+-metalloproteinase (38.5%), serine proteinase (19.5%), disintegrin (17.8%), C-type lectin-like (13.2%), PLA_2 (4.3%), Kunitz-type inhibitor (4.1%), cystatin (1.7%), and unknown (0.9%). BA-5A could not be detected in the venom proteome of Bitis arietans . The occurrence of this very low-abundance (< 0.05%) or nonexpressed disintegrin transcript indicates a hitherto unrecognized structural diversity of this protein family. Whether BA-5A plays a physiological role or represents an orphan protein which could eventually evolve a role in the adaptation of snakes to changing ecological niches and prey habits deserves further investigation.
Rod D. Wittenberg - One of the best experts on this subject based on the ideXlab platform.
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Recovering the evolutionary history of Africa’s most diverse viper genus: morphological and molecular phylogeny of Bitis (Reptilia: Squamata: Viperidae)
Organisms Diversity & Evolution, 2015Co-Authors: Rod D. Wittenberg, Robert C. Jadin, Allyson M. Fenwick, Ronald L. GutberletAbstract:Assessing evolutionary relationships among wide-ranging species can be particularly beneficial to our understanding of speciation patterns and biogeography of taxa, with broad implications for conservation and applications for human health. Integrative phylogenetic analyses that incorporate multiple independent datasets (e.g., DNA, protein, phenotype) can resolve many problematic issues in systematics such as cryptic diversity and incongruence between datasets. Vipers in the genus Bitis are widely distributed throughout much of sub-Saharan Africa, filling a variety of ecological niches and presenting an important public health problem. However, evolutionary relationships among this medically and ecologically important genus have not been fully resolved due to inadequate taxon sampling and lack of informative characters. Here, we conduct the first phylogenetic study incorporating complete sampling of known species within the genus Bitis . Using morphological, molecular, and combined approaches under multiple criteria, we recovered many of the species groups detected by previous investigators, further validating four currently recognized subgenera. Bitis arietans and Bitis worthingtoni appear to be early-diverging, monotypic lineages, while the “big Bitis ” group and the small southern African species form distinct clades. Although our study provides additional information regarding the interspecific relationships within Bitis , the placement of Bitis albanica , Bitis heraldica , and Bitis inornata remains problematic. This study enhances our understanding of the evolutionary history of species within the genus Bitis incorporating a combined evidence approach to phylogenetics.
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recovering the evolutionary history of africa s most diverse viper genus morphological and molecular phylogeny of Bitis reptilia squamata viperidae
Organisms Diversity & Evolution, 2015Co-Authors: Rod D. Wittenberg, Robert C. Jadin, Allyson M. Fenwick, Ronald L. GutberletAbstract:Assessing evolutionary relationships among wide-ranging species can be particularly beneficial to our understanding of speciation patterns and biogeography of taxa, with broad implications for conservation and applications for human health. Integrative phylogenetic analyses that incorporate multiple independent datasets (e.g., DNA, protein, phenotype) can resolve many problematic issues in systematics such as cryptic diversity and incongruence between datasets. Vipers in the genus Bitis are widely distributed throughout much of sub-Saharan Africa, filling a variety of ecological niches and presenting an important public health problem. However, evolutionary relationships among this medically and ecologically important genus have not been fully resolved due to inadequate taxon sampling and lack of informative characters. Here, we conduct the first phylogenetic study incorporating complete sampling of known species within the genus Bitis. Using morphological, molecular, and combined approaches under multiple criteria, we recovered many of the species groups detected by previous investigators, further validating four currently recognized subgenera. Bitis arietans and Bitis worthingtoni appear to be early-diverging, monotypic lineages, while the “big Bitis” group and the small southern African species form distinct clades. Although our study provides additional information regarding the interspecific relationships within Bitis, the placement of Bitis albanica, Bitis heraldica, and Bitis inornata remains problematic. This study enhances our understanding of the evolutionary history of species within the genus Bitis incorporating a combined evidence approach to phylogenetics.