A dissertation submitted to the School of Animal, Plant and Environmental Sciences, Faculty of Science, University of the Witwatersrand, Johannesburg, South Africa in fulfilment of the requirements of the degree of Masters of Science. Johannesburg, South Africa. July, 2016. / A central goal in macroecology is understanding the factors that have shaped broad-scale patterns among extant natural communities. Diet is widely cited as a key factor involved in shaping snake diversification. Amongst serpents, the adaptive radiation of elapids is unparalleled in terms of their sheer global distribution. My study was aimed at evaluating the variation in diet and how trends evolved throughout the radiation of the elapids. Based on the published natural history data of 303 elapid species, a cluster analysis revealed seven groups of species within which diet types were similar. The broad diet clusters included snake consumption, feeding on fish, ectotherms, endotherms, lizards, a generalist diet and lastly, a terrestrial invertebrate group that was the least prevalent diet type. Maximum Parsimony, Maximum Likelihood and Stochastic Character Mapping were used to trace the ancestral dietary condition in the Elapidae. Reconstructions demonstrated that starting from an ophiophagous ancestor, elapids still largely retain their early feeding habits. Despite the emergence of novel feeding patterns in the group, the predominantly conservative diet patterns suggest that niche conservatism has shaped elapid foraging dynamics. Members of the lineage characteristically feed on elongate ectothermic prey (e.g., squamates, caecilians, eels, earthworms) possibly owing to morphologically-mediated gape constraints. Phylogenetically corrected comparative methods were used to assess the influence of differences in body size, lifestyle and global scale spatial distribution of diet evolution. While lifestyle and distribution considerably influenced how diet patterns evolved, body size was not a significant determinant of the prey types taken. The lack of body size-diet association may be because larger-bodied elapids also opportunistically supplement their diet with smaller prey items. Overall, the findings indicate that recent ecological factors (e.g., lifestyle) coupled with long established factors (phylogeny, biogeography) have shaped elapid natural history patterns. This study is the first of its kind to offer a family-level, phylogenetically-based analysis on the evolutionary and contemporary feeding dynamics of the world’s most widespread snake family. / LG2017

Thesis (Ph. D.)--University of Queensland, 2001. / Includes bibliographical references.

Recentes estudos do gênero Micrurus, o qual é popularmente conhecido como o das “cobras-corais verdadeiras”, dividem este táxon em dois grupos morfologicamente distintos, o grupo das cobras corais com anéis negros em mônades (BRM) e o grupo das cobras corais com anéis negros em tríades (BRT) e, com base na morfologia macroscópica, supõe-se que esses grupos possuam estratégias reprodutivas diferenciadas. Assim, este estudo analisa e descreve a biologia reprodutiva de machos e fêmeas da espécie de serpente de tríades Micrurus lemniscatus de algumas regiões do Brasil. Micrurus lemniscatus é um complexo, com evidências polifiléticas, sendo composto por quatro subespécies: Micrurus l. carvalhoi, Micrurus l. helleri, Micrurus l. diutius e M. l. lemniscatus que habitam Mata atlântica, Cerrado e Floresta Amazônica brasileiros. Para isto foram utilizados animais preservados em coleções herpetológicas, dos quais obteveram-se dados biométricos e merísticos. Além disso, estes espécimes foram dissecados e suas gônadas medidas e coletadas para análises morfológicas e histológicas. Outras características, relacionadas à biologia reprodutiva, como presença de dimorfismo sexual, morfologia macro e microscópica, diferenciação das gônadas durante as estações e estocagem de espermatozoides também foram estudadas. O ciclo reprodutivo de Micrurus lemniscatus carvalhoi da Mata Atlântica e Cerrado foi estudado separadamente do ciclo reprodutivo de Micrurus lemniscatus lemniscatus da Amazônia e a comparação entre eles evidencia diferenças consideráveis, assim, é provável que o habitat esteja influenciando o ciclo dessas subespécies, o que pode gerar uma barreira reprodutiva e futura especiação entre elas / Recent studies about the genus Micrurus, popularly known as “true coral snakes”, divide this taxon in two morphologically distinct groups: the group which has black rings arranged in monads (BRM) and the group which has black rings in triads (BRT) and, based on macroscopic morphology, it is assumed that these groups have different reproductive strategies. Therefore, this research analyzes and describes male and female Micrurus lemniscatus reproductive biology of some regions in Brazil. M. lemniscatus is a species complex with polyphyletic evidence, consisting of four subspecies: Micrurus l. carvalhoi, Micrurus l. helleri, Micrurus l. diutius and M. l. lemniscatus, which inhabit the Atlantic rainforest, Cerrado and Amazon rainforest. For such, animals of herpetological collections were used to obtain biometric and meristic data. In addition, these specimens were dissected and their gonads measured and collected for morphological and histological analyzes. Other features related to reproductive biology were examined, such as the presence of sexual dimorphism, macroscopic and microscopic morphology, gonadal differentiation during the seasons, and sperm storage. The reproductive cycle of Micrurus lemniscatus carvalhoi from Atlantic forest and Cerrado was studied separately from the reproductive cycle of Micrurus lemniscatus lemniscatus from the Amazon forest and the comparison between them shows considerably differences, so it is likely that the habitat is influencing the cycle of these subspecies, which can generate a reproductive barrier and future speciation between them

O soro anti-elapidico brasileiro é produzido com uma mistura de veneno de Micrurus frontalis e de M. corallinus. Estudos indicam que o soro resultante não neutraliza o veneno de algumas espécies de Micrurus. Além disso, o baixo rendimento de veneno e as dificuldades de manutenção destas serpentes em cativeiro dificultam a produção de soro. Assim, um método alternativo para a produção deste soro seria de grande valor. Estudos têm mostrado que os venenos de elapideos brasileiros contêm toxinas com um elevado grau de homologia com as de suas congêneres australianas. O presente trabalho teve como objetivo comparar inicialmente o soro brasileiro e o australiano frente ao veneno de Micrurus frontalis e M. lemniscatus. A reatividade cruzada foi testada por Western-blot e ELISA com veneno de Micrurus frontalis e a capacidade neutralizante por soroneutralização com venenos de M. frontalis, M. lemniscatus, e em parceiria com venenos de M. corallinus, M. altirostris, M. spixii, M. ibiboboca, M. fulvius, M. pyrrhocryptus , M. nigrocinctus, em camundongos. Os dados obtidos indicam nível elevado de reatividade e neutralização cruzada entre os soros. Também comparamos a imunogenicidade do veneno nativo ou irradiado, não observando diferenças nos níveis de anticorpos obtidos. As serpentes australianas utilizadas para a produção de soro são mais fáceis de manusear e produzem maiores quantidades de venenos do que as corais brasileiras. Concluímos que um soro produzido com o veneno de serpentes australianas neutraliza a atividade tóxica das Micrurus estudadas, incluindo espécies que o soro nacional não neutraliza. Assim, o uso de venenos da Austrália como imunógeno constitui alternativa viável para sanar a carência do soro nacional. / The Brazilian coral snake antivenom is produced using a mixture of Micrurus corallinus and M. frontalis venom. Several studies have shown that this serum does not neutralize the venom of some species. Furthermore, the low venom yields and the difficulties of keeping coral snakes in captivity impair antivenom production. Althought belonging to different species, coral snakes and Australian elapids share many homologous toxins in their venom. The present work aimed to compare the Brazilian and Australian antivenoms against Micrurus venom. Cross- reactivity was assayed by Western blot and ELISA, while cross-neutralization was tested in mice. Both tipes of assays indicate high levels of cross-reactivity and neutralization. We also tested the immunogenicity of native or radiation-attenuated venom and no significant difference in antibody levels was observed. The Australian snakes used for antivenom production are much easier to breed and produce higher amounts of venom that their Brazilian relatives. From this study, we conclude that a serum produced using Australian venoms neutralizes the toxicity of coral snakes venoms, including several venoms which are not neutralized by the Brazilian antivenom. Thus, the use of Australian venoms as an immunogen might solve the bottleneck of coral snake antivenom production.

O soro anti-elapidico brasileiro é produzido com uma mistura de veneno de Micrurus frontalis e de M. corallinus. Estudos indicam que o soro resultante não neutraliza o veneno de algumas espécies de Micrurus. Além disso, o baixo rendimento de veneno e as dificuldades de manutenção destas serpentes em cativeiro dificultam a produção de soro. Assim, um método alternativo para a produção deste soro seria de grande valor. Estudos têm mostrado que os venenos de elapideos brasileiros contêm toxinas com um elevado grau de homologia com as de suas congêneres australianas. O presente trabalho teve como objetivo comparar inicialmente o soro brasileiro e o australiano frente ao veneno de Micrurus frontalis e M. lemniscatus. A reatividade cruzada foi testada por Western-blot e ELISA com veneno de Micrurus frontalis e a capacidade neutralizante por soroneutralização com venenos de M. frontalis, M. lemniscatus, e em parceiria com venenos de M. corallinus, M. altirostris, M. spixii, M. ibiboboca, M. fulvius, M. pyrrhocryptus , M. nigrocinctus, em camundongos. Os dados obtidos indicam nível elevado de reatividade e neutralização cruzada entre os soros. Também comparamos a imunogenicidade do veneno nativo ou irradiado, não observando diferenças nos níveis de anticorpos obtidos. As serpentes australianas utilizadas para a produção de soro são mais fáceis de manusear e produzem maiores quantidades de venenos do que as corais brasileiras. Concluímos que um soro produzido com o veneno de serpentes australianas neutraliza a atividade tóxica das Micrurus estudadas, incluindo espécies que o soro nacional não neutraliza. Assim, o uso de venenos da Austrália como imunógeno constitui alternativa viável para sanar a carência do soro nacional. / The Brazilian coral snake antivenom is produced using a mixture of Micrurus corallinus and M. frontalis venom. Several studies have shown that this serum does not neutralize the venom of some species. Furthermore, the low venom yields and the difficulties of keeping coral snakes in captivity impair antivenom production. Althought belonging to different species, coral snakes and Australian elapids share many homologous toxins in their venom. The present work aimed to compare the Brazilian and Australian antivenoms against Micrurus venom. Cross- reactivity was assayed by Western blot and ELISA, while cross-neutralization was tested in mice. Both tipes of assays indicate high levels of cross-reactivity and neutralization. We also tested the immunogenicity of native or radiation-attenuated venom and no significant difference in antibody levels was observed. The Australian snakes used for antivenom production are much easier to breed and produce higher amounts of venom that their Brazilian relatives. From this study, we conclude that a serum produced using Australian venoms neutralizes the toxicity of coral snakes venoms, including several venoms which are not neutralized by the Brazilian antivenom. Thus, the use of Australian venoms as an immunogen might solve the bottleneck of coral snake antivenom production.

Recentes estudos do gênero Micrurus, o qual é popularmente conhecido como o das “cobras-corais verdadeiras”, dividem este táxon em dois grupos morfologicamente distintos, o grupo das cobras corais com anéis negros em mônades (BRM) e o grupo das cobras corais com anéis negros em tríades (BRT) e, com base na morfologia macroscópica, supõe-se que esses grupos possuam estratégias reprodutivas diferenciadas. Assim, este estudo analisa e descreve a biologia reprodutiva de machos e fêmeas da espécie de serpente de tríades Micrurus lemniscatus de algumas regiões do Brasil. Micrurus lemniscatus é um complexo, com evidências polifiléticas, sendo composto por quatro subespécies: Micrurus l. carvalhoi, Micrurus l. helleri, Micrurus l. diutius e M. l. lemniscatus que habitam Mata atlântica, Cerrado e Floresta Amazônica brasileiros. Para isto foram utilizados animais preservados em coleções herpetológicas, dos quais obteveram-se dados biométricos e merísticos. Além disso, estes espécimes foram dissecados e suas gônadas medidas e coletadas para análises morfológicas e histológicas. Outras características, relacionadas à biologia reprodutiva, como presença de dimorfismo sexual, morfologia macro e microscópica, diferenciação das gônadas durante as estações e estocagem de espermatozoides também foram estudadas. O ciclo reprodutivo de Micrurus lemniscatus carvalhoi da Mata Atlântica e Cerrado foi estudado separadamente do ciclo reprodutivo de Micrurus lemniscatus lemniscatus da Amazônia e a comparação entre eles evidencia diferenças consideráveis, assim, é provável que o habitat esteja influenciando o ciclo dessas subespécies, o que pode gerar uma barreira reprodutiva e futura especiação entre elas / Recent studies about the genus Micrurus, popularly known as “true coral snakes”, divide this taxon in two morphologically distinct groups: the group which has black rings arranged in monads (BRM) and the group which has black rings in triads (BRT) and, based on macroscopic morphology, it is assumed that these groups have different reproductive strategies. Therefore, this research analyzes and describes male and female Micrurus lemniscatus reproductive biology of some regions in Brazil. M. lemniscatus is a species complex with polyphyletic evidence, consisting of four subspecies: Micrurus l. carvalhoi, Micrurus l. helleri, Micrurus l. diutius and M. l. lemniscatus, which inhabit the Atlantic rainforest, Cerrado and Amazon rainforest. For such, animals of herpetological collections were used to obtain biometric and meristic data. In addition, these specimens were dissected and their gonads measured and collected for morphological and histological analyzes. Other features related to reproductive biology were examined, such as the presence of sexual dimorphism, macroscopic and microscopic morphology, gonadal differentiation during the seasons, and sperm storage. The reproductive cycle of Micrurus lemniscatus carvalhoi from Atlantic forest and Cerrado was studied separately from the reproductive cycle of Micrurus lemniscatus lemniscatus from the Amazon forest and the comparison between them shows considerably differences, so it is likely that the habitat is influencing the cycle of these subspecies, which can generate a reproductive barrier and future speciation between them

Thesis (Ph. D.)--Lehigh University, 2003. / Includes vita. Includes bibliographical references (leaves 147-160).

Mini-dissertation submitted in partial compliance with the requirements of the Master’s Degree in Technology: Homoeopathy in the Faculty of Health Sciences at the Durban University of Technology, 2008. / The aim of this study was to elicit and document the effects, in the form of signs and symptoms produced in relatively healthy volunteers, of the venom of Hemachatus haemachatus, prepared in accordance with the methods set out in the homoeopathic pharmacopoeia. These signs and symptoms form the indications for the prescription of the remedy, according to the homoeopathic Law of Similars. A further aim of this study was a comparative analysis of symptoms produced by Hemachatus haemachatus 30ch with existing remedies derived from snake venom used in homeopathy, with the aim of highlighting the similarities and differences between them. The homoeopathic drug proving of Hemachatus haemachatus 30ch took the form of a double-blind, placebo controlled trial. The proving population consisted of 30 healthy subjects who met with the necessary inclusion criteria (Appendix B). Eighty percent (24 subjects) served as the experimental group, receiving the active verum in a randomised manner, while twenty percent (6 subjects) formed the placebo group, receiving non-medicated placebo powders in a randomised manner. Provers were unaware of either the nature or potency of the substance. Verum and placebo were indistinguishable from each other, and neither researcher nor volunteers knew who received verum and who received placebo. Intra-individual control was achieved through a pre-proving observation period of a week’s duration, during which provers recorded the signs and symptoms of their normal state. This symptom picture served as a baseline for comparison with symptoms noted after administration of the remedy. Verum and placebo were dispensed in the form of six powders to be taken sublingually three times a day for a period of two days, or until the onset of symptoms. Data was primarily collected in the form of a diary or journal kept by each prover in which they recorded symptoms on a daily basis. Provers were closely monitored by the researchers during this period. Data collected by the researchers during daily telephonic contacts, as well as during the pre-proving consultation, was also considered. Information obtained from the journals was then assessed by the researchers for suitability for inclusion in the materia medica of Hemachatus haemachatus. The data did not require statistical analysis. In a concurrent study of similar methodology, Cahill (2008) conducted a comparison of the symptom complex produced in the proving of Hemachatus haemachatus, with other homeopathic remedies which scored highest on repertorisation. Symptoms from both studies were collated and included in the materia medica and repertory of Hemachatus haemachatus. The investigation supported the hypothesis that Hemachatus haemachatus would produce clear and observable signs and symptoms in healthy proving volunteers. During the course of this study provers experienced a wide range of mental, emotional and physical symptoms. The highest number of symptoms was produced on the mental and emotional level. Provers experienced elation, joy, increased confidence, enthusiasm and energy. On the other hand, there was lack of confidence, vulnerability, anxiety, decreased motivation, decreased concentration, lethargy, depression and indifference. There were sudden changes in mood and provers experienced marked irritability. On the physical level, many provers experienced headaches, irritation of the eyes, symptoms resembling allergic rhinitis, sore throats, nausea, heartburn, abdominal pain and flatulence, menstrual disturbances, lumbar pain, rheumatic joint pains, and skin eruptions. There were disturbances in normal sleep patterns, subjective perceptions of increased body temperature with hot flushes, and generalised tiredness and lethargy. Symptoms obtained from the proving of Hemachatus haemachatus were analysed as part of a comparative study with other remedies derived from snake venom: Lachesis muta, Naja tripudians, Elaps corallinus, Naja mossambica and Bitis arietans arietans. This comparison highlighted both the similarities and differences between these remedies and Hemachatus haemachatus. / M

Mini-dissertation submitted in partial compliance with the requirements of the Master’s Degree in Technology: Homoeopathy in the Faculty of Health Sciences at the Durban University of Technology, 2008. / The aim of this study was to elicit and document the effects, in the form of signs and symptoms produced in relatively healthy volunteers, of the venom of Hemachatus haemachatus, prepared in accordance with the methods set out in the homoeopathic pharmacopoeia. These signs and symptoms form the indications for the prescription of the remedy, according to the homoeopathic Law of Similars. A further aim of this study was a comparative analysis of symptoms produced by Hemachatus haemachatus 30ch with existing remedies derived from snake venom used in homeopathy, with the aim of highlighting the similarities and differences between them. The homoeopathic drug proving of Hemachatus haemachatus 30ch took the form of a double-blind, placebo controlled trial. The proving population consisted of 30 healthy subjects who met with the necessary inclusion criteria (Appendix B). Eighty percent (24 subjects) served as the experimental group, receiving the active verum in a randomised manner, while twenty percent (6 subjects) formed the placebo group, receiving non-medicated placebo powders in a randomised manner. Provers were unaware of either the nature or potency of the substance. Verum and placebo were indistinguishable from each other, and neither researcher nor volunteers knew who received verum and who received placebo. Intra-individual control was achieved through a pre-proving observation period of a week’s duration, during which provers recorded the signs and symptoms of their normal state. This symptom picture served as a baseline for comparison with symptoms noted after administration of the remedy. Verum and placebo were dispensed in the form of six powders to be taken sublingually three times a day for a period of two days, or until the onset of symptoms. Data was primarily collected in the form of a diary or journal kept by each prover in which they recorded symptoms on a daily basis. Provers were closely monitored by the researchers during this period. Data collected by the researchers during daily telephonic contacts, as well as during the pre-proving consultation, was also considered. Information obtained from the journals was then assessed by the researchers for suitability for inclusion in the materia medica of Hemachatus haemachatus. The data did not require statistical analysis. In a concurrent study of similar methodology, Cahill (2008) conducted a comparison of the symptom complex produced in the proving of Hemachatus haemachatus, with other homeopathic remedies which scored highest on repertorisation. Symptoms from both studies were collated and included in the materia medica and repertory of Hemachatus haemachatus. The investigation supported the hypothesis that Hemachatus haemachatus would produce clear and observable signs and symptoms in healthy proving volunteers. During the course of this study provers experienced a wide range of mental, emotional and physical symptoms. The highest number of symptoms was produced on the mental and emotional level. Provers experienced elation, joy, increased confidence, enthusiasm and energy. On the other hand, there was lack of confidence, vulnerability, anxiety, decreased motivation, decreased concentration, lethargy, depression and indifference. There were sudden changes in mood and provers experienced marked irritability. On the physical level, many provers experienced headaches, irritation of the eyes, symptoms resembling allergic rhinitis, sore throats, nausea, heartburn, abdominal pain and flatulence, menstrual disturbances, lumbar pain, rheumatic joint pains, and skin eruptions. There were disturbances in normal sleep patterns, subjective perceptions of increased body temperature with hot flushes, and generalised tiredness and lethargy. Symptoms obtained from the proving of Hemachatus haemachatus were analysed as part of a comparative study with other remedies derived from snake venom: Lachesis muta, Naja tripudians, Elaps corallinus, Naja mossambica and Bitis arietans arietans. This comparison highlighted both the similarities and differences between these remedies and Hemachatus haemachatus.

Thesis (M.Tech.: Homoeopathy)--Durban University of Technology, 2008 / The proving substance Hemachatus haemachatus commonly known as the Rinkhals belongs to the family of Elapidae. This spitting-cobra is a local snake found only in Southern Africa. This proving tested the effects of the thirtieth centesimal (30CH) potency of venom from Hemachatus haemachatus on healthy provers. OBJECTIVES It was hypothesised that Hemachatus haemachatus 30CH would produce clearly observable signs and symptoms in healthy provers, and that the comparison of Hemachatus haemachatus to those yielding the highest numerical value and total number of rubrics on repertorisation of the proving symptoms would highlight differences and similarities between the remedy symptoms so that confusion as to the indication is eliminated. It was hypothesised that a fuller understanding of Hemachatus haemachatus and its relationship to other remedies would be gained following this comparison. METHODOLOGY A double blind, placebo controlled proving of Hemachatus haemachatus 30CH was conducted on thirty healthy volunteers who met the inclusion criteria. Six of these thirty provers randomly received placebo, with neither prover nor researcher knowing whom received placebo. Provers had a homoeopathic case history taken and a physical examination performed on them prior to commencement of the proving. The provers recorded their signs and symptoms 6 by means of a journal before, during and after administration of the remedy. On completion of the proving, the information obtained was correlated and assessed by the two researchers, De la Rouviere and Cahill. The symptoms elicited during the proving were translated into materia medica and repertory language, and a homoeopathic picture of the remedy was subsequently formulated. Data from the case histories, physical examinations and group discussions were also considered in the assessment. RESULTS During the period of investigation, provers experienced a variety of symptoms on the mental, emotional and physical spheres. On the mental emotional sphere there was a marked degree of irritability and changeability in moods as is commonly seen in many of the snake remedies. Along with this, it was noted that there were feelings of anxiety for reasons unknown, a sense of having lost something or someone close, and a desire to be left alone. There were also a great number of feelings regarding the home, where there were feelings of the home being a place of safety and wanting order in the home. On a physical level, many of the provers noted varying degrees of abdominal discomfort and headaches. Along with anxiety, provers experienced palpitations and sensations of chest restriction or constriction with shortness of breath. There were a variety of musculoskeletal symptoms ranging from painful joints in the fingers to stiffness and tightness in the neck and back. Provers noted flushes of heat and alterations of their internal thermostat. Provers experienced marked dryness of the mucus membranes and the skin, and there was also a general feeling of weakness and heaviness as well as a marked aggravation in the mornings on waking. 7 CONCLUSIONS Symptoms obtained from the proving of Hemachatus haemachatus 30CH were studied and evaluated. Those symptoms that appeared to represent the remedy picture of Hemachatus haemachatus most accurately in the researchers‟ opinion were used in the repertorisation of the remedy. The investigation supported the hypothesis that Hemachatus haemachatus 30CH would produce clearly observable signs and symptoms in healthy provers. The subsequent comparison of the proving symptoms of Hemachatus haemachatus to Lycopodium (Club moss), Sulphur, Alumina (Aluminium oxide), Sepia (Cuttle fish) and Calcarea carbonica (Carbonate of Lime) highlighted differences and similarities between these remedies and Hemachatus haemachatus. The further comparison of remedies that came up on repertorisation restricted to the plant, mineral and animal kingdoms respectively provided a further comparison of remedies, which aimed at enhancing the differentiation of Hemachatus haemachatus to other similar remedies. / M