Amphibians are regarded as sensitive indicators of environmental change and are therefore excellent subjects for use in ecotoxicology. The Phongolo River floodplain is South Africa’s most diverse natural floodplain system and hosts more than 40 frog species. It is also a malaria endemic region and is subjected to active spraying with Dichlorodiphenyltrichloroethane (DDT) through means of indoor residual spraying over the summer months. The upper Phongolo River runs through agricultural landscape and is subjected to runoff from forest plantations, orchards and sugar cane plantations. In this study residue levels of 22 different organochlorine pesticides (OCPs) were analysed in selected amphibian species from in and around the Ndumo Nature Reserve coupled with 12 different biomarker response assays to determine environmental exposure levels and possible sublethal effects in amphibians from the lower Phongolo River floodplain. Seasonal change, direct influence of anthropogenic activity and the influence of species’ aquatic preference in habitat selection were all factors considered during this assessment. Stable Isotope analyses were performed on 11 different food web components In order to determine the food web structure pertaining to Xenopus muelleri (Müller's platanna). Samples were collected during both high and low flow seasons from inside and outside Ndumo Nature Reserve. Organochlorine pesticide bioaccumulation was analysed in whole frog samples using a GC-μECD. Results indicated significant seasonal variation in OCP levels and exposure composition. Significant differences between inside and outside sites were also noted. Dichlorodiphenyltrichloroethane in its different isomer forms and their metabolites along with the hexachlorocyclohexane (HCH) isomers was the two main contributing OCP groups detected. Total OCP levels from all sample sets ranged between 8.71 ng/g lipid and 21,399.03 ng/g lipid. An increase in OCP accumulation was observed for X. muelleri over a period of one year. Organochlorine pesticides are known to have neurotoxic effects causing imbalances in Na+, K+, and Ca+ ion exchange. Hyperactivity has been reported in Rana temporaria (European Common frog) tadpoles exposed to p,p-DDT concentrations above 110,000 ng/g lipid. Despite OCP levels measured in frogs from this study being lower than reported toxic levels, the biomarker response assays indicated definite oxidative stress responses correlating to OCP bioaccumulation, with other minor responses shown. Cellular energy allocation showed a shift in the main energy source type from proteins to lipids correlating to increased OCP bioaccumulation. A slight inhibition response was noted in the hepato-somatic index correlating to γ-HCH bioaccumulation. Stable isotope analyses indicated food web structure differences between inside and outside the reserve, with outside showing less clear distinction between trophic groups and nitrogen enrichment of primary producers. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Amphibians are regarded as sensitive indicators of environmental change and are therefore excellent subjects for use in ecotoxicology. The Phongolo River floodplain is South Africa’s most diverse natural floodplain system and hosts more than 40 frog species. It is also a malaria endemic region and is subjected to active spraying with Dichlorodiphenyltrichloroethane (DDT) through means of indoor residual spraying over the summer months. The upper Phongolo River runs through agricultural landscape and is subjected to runoff from forest plantations, orchards and sugar cane plantations. In this study residue levels of 22 different organochlorine pesticides (OCPs) were analysed in selected amphibian species from in and around the Ndumo Nature Reserve coupled with 12 different biomarker response assays to determine environmental exposure levels and possible sublethal effects in amphibians from the lower Phongolo River floodplain. Seasonal change, direct influence of anthropogenic activity and the influence of species’ aquatic preference in habitat selection were all factors considered during this assessment. Stable Isotope analyses were performed on 11 different food web components In order to determine the food web structure pertaining to Xenopus muelleri (Müller's platanna). Samples were collected during both high and low flow seasons from inside and outside Ndumo Nature Reserve. Organochlorine pesticide bioaccumulation was analysed in whole frog samples using a GC-μECD. Results indicated significant seasonal variation in OCP levels and exposure composition. Significant differences between inside and outside sites were also noted. Dichlorodiphenyltrichloroethane in its different isomer forms and their metabolites along with the hexachlorocyclohexane (HCH) isomers was the two main contributing OCP groups detected. Total OCP levels from all sample sets ranged between 8.71 ng/g lipid and 21,399.03 ng/g lipid. An increase in OCP accumulation was observed for X. muelleri over a period of one year. Organochlorine pesticides are known to have neurotoxic effects causing imbalances in Na+, K+, and Ca+ ion exchange. Hyperactivity has been reported in Rana temporaria (European Common frog) tadpoles exposed to p,p-DDT concentrations above 110,000 ng/g lipid. Despite OCP levels measured in frogs from this study being lower than reported toxic levels, the biomarker response assays indicated definite oxidative stress responses correlating to OCP bioaccumulation, with other minor responses shown. Cellular energy allocation showed a shift in the main energy source type from proteins to lipids correlating to increased OCP bioaccumulation. A slight inhibition response was noted in the hepato-somatic index correlating to γ-HCH bioaccumulation. Stable isotope analyses indicated food web structure differences between inside and outside the reserve, with outside showing less clear distinction between trophic groups and nitrogen enrichment of primary producers. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Na Colômbia ocorrem 787 espécies de anfíbios. Por causa da preocupação com o estado de conservação de muitas dessas espécies, tem se sugerido que na Colômbia deveriam-se priorizar as pesquisas em taxonomia e ecologia em regiões sub-amostradas a fim de intensificar o conhecimento e conservação dos anfíbios colombianos. Baseados em uma análise cienciométrica de 319 trabalhos sobre a ecologia dos anfíbios colombianos publicados entre 1840 e 2014 (No Capítulo 1), identificamos as tendências nos esforços realizados em distintos temas de pesquisa, e a distribuição regional e taxonômica desses estudos. A maioria dos estudos (67%) foi realizada na região Andina colombiana em comparação com outras regiões naturais da Colômbia. Apenas 46% das espécies de anfíbios ocorrendo na Colômbia foi tratada nos estudos analizados, e a maioria (58%) delas é da região Andina. Entre as publicações analizadas identificamos 14 temas de pesquisa em ecologia, dos quais ecologia reprodutiva (26%), conservação de espécies (23%) e dieta (14%) foram os mais pesquisados. Nossos dados mostraram que na Colômbia há um considerável avanço na pesquisa sobre a ecologia dos anfíbios do país, mas ainda são necessários esforços para cobrir muitos vazios de informação para muitas regiões e para muitas espécies de anfíbios que possuem dados incipientes. No sudoeste da Cordilheira Ocidental colombiana há pouca informação ecológica sobre os anfíbios ali ocorrendo. A fim de saber alguns aspectos ecológicos dessas espécies, desenvolvimos três estudos sobre a diversidade e ecologia de anfíbios presentes na Reserva Natural Río Ñambí (a seguir RNRÑ). No Capítulo 2 apresentamos uma análise sistemática do gênero Andinophryne (Família Bufonidae), composto por três espécies, A. atelopoides, A. colomai (presente na RNRÑ) e A. olallai. As filogenias mostraram que Andinophryne está incorporado dentro de Rhaebo. Portanto, sinonimizamos Andinophryne sob Rhaebo e discutimos as sinapomorfias morfológicas putativas para Rhaebo. Além, fornecemos informações ecológicas e sobre o estado de conservação das três espécies incluídas na nova combinação taxonômica. No Capítulo 3 apresentamos uma lista de 19 espécies de anfíbios pertencentes a oito famílias, com uma dominância numérica da família Craugastoridae e do gênero Pristimantis. As espécies com a maior abundância relativa (> 25%) foram Pristimantis labiosus e P. verecundus. Sete diferentes modos de reprodução foram reconhecidos, com a maioria das espécies (68%) possuindo desenvolvimento direto de ovos. Cinco (26%) das espécies registradas estão classificadas dentro das categorias de maior ameaça de extinção. Reportamos para sete espécies a extensão da faixa de distribuição geográfica latitudinal na Colômbia. No Capítulo 4 comparamos a dieta de jovens e adultos de P. labiosus para identificar se houve uma mudança ontogenética no tamanho de presa consumido com o aumento na largura da boca. A dieta foi composta por 19 categorias de presas (> artrópodes), com as duas classes de idade consumindo um similar espectro de categorias. Os jovens têm um nicho trófico maior (0,45) do que os adultos (0,25), com uma sobreposição de nicho relativamente baixa (0,39) entre eles. Apesar da diferencia na largura da boca entre jovens e adultos, não houve uma correspondente mudança ontogenética no tamanho de presa consumida. Consideramos P. labiosus como um predador generalista que parece consumer uma ampla gama de tipos e tamanhos de presas / Colombia harbors 787 species of amphibians. Because of concerns with the conservation status of many of these species, it has been suggested that in Colombia should prioritize research in taxonomy and ecology in undersampled regions in order to enhance the knowledge and conservation of Colombian amphibians. Based on a scientometric analysis of 319 works on ecology of Colombian amphibians published between 1840 and 2014 (Chapter 1), we identified trends in the efforts made in different research topics, and both regional and taxonomic distribution of these studies. Most studies (67%) was conducted in the Colombian Andean region compared to other natural regions of Colombia. Only 46% of amphibian species occurring in Colombia was included in the reviewed studies, and the most of them (58%) is from the Andean region. Among the reviewed publications, we identified 14 research topics in ecology, of which reproductive ecology (26%), species conservation (23%), and diet (14%) were the most investigated. Our data showed that in Colombia, there is a considerable advance in research on amphibian ecology, but we need further efforts to fill many gaps in information for many regions and for many amphibian species possessing incipient data. In southwestern of the Colombian western Andes there is little information about the species occurring there. In order to know some ecological aspects of these species, we conducted three studies on diversity and ecology of the amphibians present in the Reserva Natural Río Ñambí (hereafter RNRÑ). In the Chapter 2, we show an analysis of the systematics of the genus Andinophryne (Family Bufonidae), composed by three species, A. atelopoides, A. colomai (present in the RNRÑ) and A. olallai. The phylogenies showed that Andinophryne is embedded within Rhaebo. Therefore, we synonymize Andinophryne under Rhaebo and we discussed the putative morphological synapomorphies for Rhaebo. In addition, we provide ecological informations and on the conservation status of the three species included in the new taxonomic combination. In the Chapter 3, we show a list of 19 amphibian species from eight families, with a numerical dominance of the Family Craugastoridae and of the genus Pristimantis. The species with the highest relative abundance (>25%) were Pristimantis labiosus and P. verecundus. Seven different reproductive modes were recognized, with most species (68%) presented direct development of eggs. Five (26%) of the species recorded are classified in the categories of greatest threat of extinction. We report for seven species an extension of the latitudinal distribution range in Colombia. In the Chapter 4, we compared the diet of juveniles and adults of P. labiosus in order to identify if there was an ontogenetic shift in the prey size consumed as mouth width increases. The diet was composed of 19 prey categories (> arthropods), and the two age-classes consumed a similar array of prey categories. Juveniles had a broader trophic niche (0.57) than adults (0.25), with a relatively low trophic overlap among them (0.39). Although we found differences in mouth width between juveniles and adults, we did not observe a corresponding ontogenetic shift in prey size consumption. We consider that P. labiosus is a generalist predator, consuming a wide range of prey types and sizes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Na Colômbia ocorrem 787 espécies de anfíbios. Por causa da preocupação com o estado de conservação de muitas dessas espécies, tem se sugerido que na Colômbia deveriam-se priorizar as pesquisas em taxonomia e ecologia em regiões sub-amostradas a fim de intensificar o conhecimento e conservação dos anfíbios colombianos. Baseados em uma análise cienciométrica de 319 trabalhos sobre a ecologia dos anfíbios colombianos publicados entre 1840 e 2014 (No Capítulo 1), identificamos as tendências nos esforços realizados em distintos temas de pesquisa, e a distribuição regional e taxonômica desses estudos. A maioria dos estudos (67%) foi realizada na região Andina colombiana em comparação com outras regiões naturais da Colômbia. Apenas 46% das espécies de anfíbios ocorrendo na Colômbia foi tratada nos estudos analizados, e a maioria (58%) delas é da região Andina. Entre as publicações analizadas identificamos 14 temas de pesquisa em ecologia, dos quais ecologia reprodutiva (26%), conservação de espécies (23%) e dieta (14%) foram os mais pesquisados. Nossos dados mostraram que na Colômbia há um considerável avanço na pesquisa sobre a ecologia dos anfíbios do país, mas ainda são necessários esforços para cobrir muitos vazios de informação para muitas regiões e para muitas espécies de anfíbios que possuem dados incipientes. No sudoeste da Cordilheira Ocidental colombiana há pouca informação ecológica sobre os anfíbios ali ocorrendo. A fim de saber alguns aspectos ecológicos dessas espécies, desenvolvimos três estudos sobre a diversidade e ecologia de anfíbios presentes na Reserva Natural Río Ñambí (a seguir RNRÑ). No Capítulo 2 apresentamos uma análise sistemática do gênero Andinophryne (Família Bufonidae), composto por três espécies, A. atelopoides, A. colomai (presente na RNRÑ) e A. olallai. As filogenias mostraram que Andinophryne está incorporado dentro de Rhaebo. Portanto, sinonimizamos Andinophryne sob Rhaebo e discutimos as sinapomorfias morfológicas putativas para Rhaebo. Além, fornecemos informações ecológicas e sobre o estado de conservação das três espécies incluídas na nova combinação taxonômica. No Capítulo 3 apresentamos uma lista de 19 espécies de anfíbios pertencentes a oito famílias, com uma dominância numérica da família Craugastoridae e do gênero Pristimantis. As espécies com a maior abundância relativa (> 25%) foram Pristimantis labiosus e P. verecundus. Sete diferentes modos de reprodução foram reconhecidos, com a maioria das espécies (68%) possuindo desenvolvimento direto de ovos. Cinco (26%) das espécies registradas estão classificadas dentro das categorias de maior ameaça de extinção. Reportamos para sete espécies a extensão da faixa de distribuição geográfica latitudinal na Colômbia. No Capítulo 4 comparamos a dieta de jovens e adultos de P. labiosus para identificar se houve uma mudança ontogenética no tamanho de presa consumido com o aumento na largura da boca. A dieta foi composta por 19 categorias de presas (> artrópodes), com as duas classes de idade consumindo um similar espectro de categorias. Os jovens têm um nicho trófico maior (0,45) do que os adultos (0,25), com uma sobreposição de nicho relativamente baixa (0,39) entre eles. Apesar da diferencia na largura da boca entre jovens e adultos, não houve uma correspondente mudança ontogenética no tamanho de presa consumida. Consideramos P. labiosus como um predador generalista que parece consumer uma ampla gama de tipos e tamanhos de presas / Colombia harbors 787 species of amphibians. Because of concerns with the conservation status of many of these species, it has been suggested that in Colombia should prioritize research in taxonomy and ecology in undersampled regions in order to enhance the knowledge and conservation of Colombian amphibians. Based on a scientometric analysis of 319 works on ecology of Colombian amphibians published between 1840 and 2014 (Chapter 1), we identified trends in the efforts made in different research topics, and both regional and taxonomic distribution of these studies. Most studies (67%) was conducted in the Colombian Andean region compared to other natural regions of Colombia. Only 46% of amphibian species occurring in Colombia was included in the reviewed studies, and the most of them (58%) is from the Andean region. Among the reviewed publications, we identified 14 research topics in ecology, of which reproductive ecology (26%), species conservation (23%), and diet (14%) were the most investigated. Our data showed that in Colombia, there is a considerable advance in research on amphibian ecology, but we need further efforts to fill many gaps in information for many regions and for many amphibian species possessing incipient data. In southwestern of the Colombian western Andes there is little information about the species occurring there. In order to know some ecological aspects of these species, we conducted three studies on diversity and ecology of the amphibians present in the Reserva Natural Río Ñambí (hereafter RNRÑ). In the Chapter 2, we show an analysis of the systematics of the genus Andinophryne (Family Bufonidae), composed by three species, A. atelopoides, A. colomai (present in the RNRÑ) and A. olallai. The phylogenies showed that Andinophryne is embedded within Rhaebo. Therefore, we synonymize Andinophryne under Rhaebo and we discussed the putative morphological synapomorphies for Rhaebo. In addition, we provide ecological informations and on the conservation status of the three species included in the new taxonomic combination. In the Chapter 3, we show a list of 19 amphibian species from eight families, with a numerical dominance of the Family Craugastoridae and of the genus Pristimantis. The species with the highest relative abundance (>25%) were Pristimantis labiosus and P. verecundus. Seven different reproductive modes were recognized, with most species (68%) presented direct development of eggs. Five (26%) of the species recorded are classified in the categories of greatest threat of extinction. We report for seven species an extension of the latitudinal distribution range in Colombia. In the Chapter 4, we compared the diet of juveniles and adults of P. labiosus in order to identify if there was an ontogenetic shift in the prey size consumed as mouth width increases. The diet was composed of 19 prey categories (> arthropods), and the two age-classes consumed a similar array of prey categories. Juveniles had a broader trophic niche (0.57) than adults (0.25), with a relatively low trophic overlap among them (0.39). Although we found differences in mouth width between juveniles and adults, we did not observe a corresponding ontogenetic shift in prey size consumption. We consider that P. labiosus is a generalist predator, consuming a wide range of prey types and sizes