As doenças de veiculação hídrica são uma preocupação no mundo todo, e tem aumentado cada vez mais o interesse nos estudos relacionados aos protozoários Cryptosporidium spp. e Giardia spp.. A presença de protozoários em águas de abastecimento está vinculada ao lançamento de esgoto nos mananciais, de modo que o tratamento de esgotos sanitários é de extrema importância para reduzir o risco de surtos de protozooses. Este trabalho avaliou a ocorrência e a possibilidade de remoção de cistos de Giardia spp. e oocistos de Cryptosporidium spp. em dois reatores UASB, um piloto e um em escala plena, operados com esgoto sanitário e um TDH de aproximadamente 8h, e um reator UASB piloto, operado com água negra simulada e um TDH aproximado de três dias. Nos dois reatores operados com esgoto sanitário as remoções foram semelhantes. A média de DQO total afluente, no reator piloto e no de escala plena, foi de 271 e 410 mg/L, respectivamente, e as remoções médias de DQO foram de 50,62 e 45,80%, respectivamente. Com relação aos exames parasitológicos, quase não houve remoção em ambos os reatores e, na maior parte dos casos, o número de (oo)cistos detectados no efluente foi maior do que no afluente. Já no reator piloto operado com águas negras, foram observadas remoções mais elevadas. A DQO total afluente desse reator variou de 837 a 3269 mg/L e a remoção média observada foi de 71,22%. Neste reator também foram observadas maiores remoções de (oo)cistos, sendo que a remoção de cistos Giardia spp. variou de 67,73 a 93,69% e, em um dos ensaios, foi observada uma remoção de 70% de oocistos de Cryptosporidium spp. Acredita-se que o melhor desempenho desse reator se deve ao seu elevado tempo de detenção hidráulico. Foi avaliada, ainda, a ocorrência de (oo)cistos no lodo dos três reatores, resultando em valores bastante elevados, da ordem de 105 cistos/g de massa seca, em todos eles. De qualquer modo, em todos os reatores avaliados, mesmo o que apresentou as maiores eficiências, o efluente ainda apresentava altas concentrações de poluentes, necessitando de um pós-tratamento para a remoção dos remanescentes de matéria orgânica, sólidos e patógenos. / The waterborne diseases are a concern worldwide, and has grown more and more interest in studies related to the protozoa Cryptosporidium spp. and Giardia spp. The presence of protozoa in drinking water is related to the release of sewage in water sources. So that, the sewage treatment is extremely important to reduce the risk of outbreaks protozoa. This study evaluated the occurrence and the possibility of removal of Giardia spp. and Cryptosporidium spp. in two UASB reactors, a pilot and a full-scale, operated with sewage and a HRT of 8 hours, approximately, and a pilot UASB reactor, operated with simulated black water and an approximate HRT of three days. Removals were similar in both reactors operated with sewage. The average influent total COD, in pilot and full scale reactors, was 271 and 410 mg/L, respectively, and the average COD removal were 50.62 and 45.80%, respectively. With the parasitological test, there was almost no removal in both reactors and, in most cases, the number of (oo)cysts detected in the effluent was higher than in the influent. In the pilot reactor operated with black water, higher removals were observed. The total COD influent of this reactor ranged from 837 to 3269 mg/L and the average removal observed was 71.22%. In this reactor (oo)cysts removals were also higher, and the removal of Giardia cysts ranged from 67.73 to 93.69%, and in one of the trials, a 70% removal of Cryptosporidium spp was observed. It is believed that the best performance of this reactor is due to its high hydraulic retention time. The occurrence of (oo)cysts in the sludge of the three reactors was also evaluated, resulting in very high values of the order of 105 cysts/g of dry weight, in all of them. Anyway, in all this reactor, even the one with the highest efficiency, the effluent still had high concentrations of pollutants, necessitating a post treatment for the removal of remaining organic matter, solids and pathogens.

Esgotos urbanos constituem compartimentos ambientais que favorecem a emergência e re-emergência enfermidades de veiculação hídrica. Este estudo objetivou analisar cistos de Giardia spp., oocistos de Cryptosporidium spp., assim como cistos, ovos e larvas de outros protozoários e helmintos patogênicos; Coliformes Totais e Termotolerantes, Rotavírus e Adenovírus em amostras de esgoto bruto e tratado da Estação de Tratamento de Esgoto de Ribeirão Preto - SP. As coletas de esgoto foram realizadas no ponto de entrada e no ponto de saída da ETE-Ribeirão Preto/SP. As análises de cistos de Giardia spp. e oocistos de Cryptosporidium spp. foram realizados pelo Método 1623 da EPA. Ovos, cistos e larvas de outros protozoários e helmintos foram analisados pelo Método de Sedimentação proposto pela CETESB (1989). A análise de coliformes totais e termotolerantes foi realizada pela Técnica de Tubos Múltiplos (Colilert®). A análise de vírus foi realizada através do Teste de Elisa com o KIT da RIDASCREEN® Enzimaimunoensaio. Os resultados obtidos revelaram que a concentração de Giardia spp. no esgoto bruto variou de 120 a 2200 cistos/L, já no esgoto tratado essa concentração variou de 0,45 a 3,5 cistos/L. Com relação aos oocistos de Cryptosporidium spp. a concentração no esgoto bruto variou de não detectável a 28,9 oocistos/L e no esgoto tratado as concentrações variaram de não detectável a 1,05 oocistos/L. O processo de lodos ativados na ETE-RP promoveu uma remoção parcial de parasitas, tais como: Endolimax nana, Entamoeba coli, Entamoeba hystolitica, Ancylostoma sp., Ascaris sp. Fasciola hepatica e Strongyloides stercoralis, cujo fator de redução variou entre 18,2 e 100%. Pode-se observar que houve diferença estatisticamente significante entre o número mais provável de coliformes totais e termotolerantes no esgoto bruto e esgoto tratado, mostrando uma redução na concentração desses organismos indicadores após o tratamento do esgoto na ETE-Ribeirão Preto/SP. Não houve correlação significativa entre as concentrações de coliformes totais e termotolerantes em relação à concentração de cistos de Giardia spp. e oocistos de Cryptosporidium spp.. A análise de vírus mostrou resposta positiva para Rotavirus e Adenovirus em todas as amostras analisadas (esgoto bruto e tratado), mostrando menor absorvância no esgoto tratado. Os resultados obtidos contribuem para a identificação e caracterização de microorganismos de veiculação hídrica no esgoto tratado na ETE-RP, fornecendo dados úteis para a definição de políticas públicas de saneamento, referentes ao controle da qualidade microbiológica do esgoto no país. / Sewage treatment is still precarious in many Brazilian cities, which has contributed to the emergence and re-emergence of waterborne diseases. This study analyzed Giardia spp. cysts and Cryptosporidium spp. oocysts, as well as cysts, eggs and larvae of other protozoa and helminthes. Also part of the study were total and thermotolerant coliforms, Rotavirus and Adenovirus in samples of raw and treated sewage of the Sewage Treatment Station (ETE) of Ribeirão Preto, SP, Brazil. Sewage samples were taken at the points of entry and exit of the ETE, Ribeirão Preto. Analyses of Giardia spp. cysts and Cryptosporidium spp. oocysts were performed through the EPA\'s method 1623. Eggs, cysts and larvae of other protozoa and helminthes were analyzed utilizing the method of sedimentation as proposed by CETESB (1989). Analyses of total and thermotolerant coliforms were performed using the Multiple Tube method (Colilert®). Viral analyses were performed through the RIDASCREEN® enzyme immunoassay ELISA test kit. The results revealed that the concentration of Giardia spp. in raw sewage varied from 120 to 2,200 cysts/L; this concentration varied from 0.45 to 3.5 cysts/L in treated sewage. The concentration of Cryptosporidium spp. oocysts in raw sewage varied from non-detectable to 28.9 oocysts/L and from non-detectable to 1.05 oocysts/L in treated sewage. The activated sludge process used in ETE promoted a partial removal of parasites such as: Endolimax nana, Entamoeba coli, Entamoeba hystolitica, Ancylostoma sp., Ascaris sp. Fasciola hepatica and Strongyloides stercoralis. Reduction factors varied from 18.2 to 100%. A statistically significant difference was observed between the most probable number of total and thermotolerant coliforms in raw and treated sewage, showing a reduction in the concentration of these indicators after the treatment of sewage in ETE, Ribeirão Preto. No statistically significant correlation was found between the concentrations of total and thermotolerant coliforms in relation to the concentration of Giardia cysts spp. and Cryptosporidium spp. oocysts. Analyzing viruses showed positive responses for Rotavirus and Adenovirus in all the studied samples (raw and treated sewage), with a smaller absorbance in the treated sewage. The results contribute to the identification and evaluation of the prevalence of waterborne microorganisms in treated sewage in ETE, Ribeirão Preto, providing useful data for the definition of public policies for sanitation concerning microbiological quality control in Brazil\'s sewage.

Giardia e Cryptosporidium são protozoários cosmopolitas cuja epidemiologia é especialmente importante devido ao seu expressivo potencial zoonótico. Animais silvestres são frequentemente relatados como reservatórios da giardiose e criptosporidiose humanas, todavia, são escassas as evidências sobre sua real importância na manutenção e disseminação destas protozooses. No intuito de avaliar a ocorrência e determinar os genótipos responsáveis pela infecção de mamíferos neotropicais, 452 amostras fecais procedentes de 52 diferentes espécies, in situ e ex situ, de sete localidades distintas foram avaliadas por métodos de diagnóstico microscópico, seguidos por técnicas moleculares de amplificação (Nested PCR), sequenciamento e caracterização genotípica. Os resultados revelaram prevalência aparente de 6,2% para Giardia spp. e de 4,8% para Cryptosporidium spp. (n=343). Dezessete diferentes espécies de mamíferos silvestres foram positivas, sendo 11 para Giardia spp., nove para Cryptosporidium spp. e três para ambos os protozoários. A caracterização molecular revelou a predominante presença de genótipos zoonóticos em mamíferos cativos (Giardia duodenalis genótipo AI) e de genótipos hospedeiro-específicos em animais de vida livre (Cryptosporidium sp. rat genotype III e Cryptosporidium wrairi). Foram também identificados Giardia duodenalis genótipo D em cachorro-do-mato Cerdocyon thous e Cryptosporidium sp. deer mouse genotype IV em bugio-preto Alouatta caraya, ambos mantidos em cativeiro. Aspectos ecológicos, como habitat, guilda trófica, estratégia do uso do ambiente e influência antrópica, foram considerados relevantes para a ocorrência dos parasitas. Tais achados demonstraram que animais silvestres podem ser infectados por genótipos zoonóticos e específicos dos agentes, o que revela a importância de estudos envolvendo esta abordagem para sugerir possíveis relações entre os protozoários, hospedeiros humanos, animais domésticos e silvestres perante diferentes características ambientais. / Giardia and Cryptosporidium are cosmopolitan protozoans whose epidemiology is especially important due to its significant zoonotic potencial. Wild animals are often reported as reservoirs of human giardiosis and cryptosporidiosis, however, there is little evidence about their real importance in the maintenance and dissemination of these protozoosis. In order to evaluate the occurrence and determine the genotypes responsible for neotropical mammals infection, 452 fecal samples of 52 different species, in situ and ex situ, of seven locations were evaluated by microscopic methods of diagnosis, followed by molecular amplification (Nested PCR), sequencing and genotypic chacacterization techniques. The results revealed an apparent prevalence of 6,2% for Giardia spp. and 4,8% for Cryptosporidium spp. (n=343). Seventeen different species of wild mammals were positive, 11 for Giardia spp., nine for Cryptosporidium spp. and three for both protozoans. Molecular characterization shows predominant presence of zoonotic genotypes in captive mammals (Giardia duodenalis genotype AI) and host-specific genotypes in free-living animals (Cryptosporidium sp. rat genotype III and Cryptosporidium wrairi). Giardia duodenalis genotype D in crab-eating fox Cerdocyon thous and Cryptosporidium sp. deer mouse genotype IV in black howler monkey Alouatta caraya, both in captivity, were also identified. Ecological aspects, like habitat, trophic guilds, strategy of using the environment and human influence, were considered relevant to occurrence of the parasites. These findings could demonstrate wild mammals can be infected by zoonotic and specific genotypes of agents, which shows the importance of studies using this approach to suggest possible relationships between protozoans, human hosts, domestic animals and wildlife facing different environmental characteristics.

Giardia e Cryptosporidium são protozoários cosmopolitas cuja epidemiologia é especialmente importante devido ao seu expressivo potencial zoonótico. Animais silvestres são frequentemente relatados como reservatórios da giardiose e criptosporidiose humanas, todavia, são escassas as evidências sobre sua real importância na manutenção e disseminação destas protozooses. No intuito de avaliar a ocorrência e determinar os genótipos responsáveis pela infecção de mamíferos neotropicais, 452 amostras fecais procedentes de 52 diferentes espécies, in situ e ex situ, de sete localidades distintas foram avaliadas por métodos de diagnóstico microscópico, seguidos por técnicas moleculares de amplificação (Nested PCR), sequenciamento e caracterização genotípica. Os resultados revelaram prevalência aparente de 6,2% para Giardia spp. e de 4,8% para Cryptosporidium spp. (n=343). Dezessete diferentes espécies de mamíferos silvestres foram positivas, sendo 11 para Giardia spp., nove para Cryptosporidium spp. e três para ambos os protozoários. A caracterização molecular revelou a predominante presença de genótipos zoonóticos em mamíferos cativos (Giardia duodenalis genótipo AI) e de genótipos hospedeiro-específicos em animais de vida livre (Cryptosporidium sp. rat genotype III e Cryptosporidium wrairi). Foram também identificados Giardia duodenalis genótipo D em cachorro-do-mato Cerdocyon thous e Cryptosporidium sp. deer mouse genotype IV em bugio-preto Alouatta caraya, ambos mantidos em cativeiro. Aspectos ecológicos, como habitat, guilda trófica, estratégia do uso do ambiente e influência antrópica, foram considerados relevantes para a ocorrência dos parasitas. Tais achados demonstraram que animais silvestres podem ser infectados por genótipos zoonóticos e específicos dos agentes, o que revela a importância de estudos envolvendo esta abordagem para sugerir possíveis relações entre os protozoários, hospedeiros humanos, animais domésticos e silvestres perante diferentes características ambientais. / Giardia and Cryptosporidium are cosmopolitan protozoans whose epidemiology is especially important due to its significant zoonotic potencial. Wild animals are often reported as reservoirs of human giardiosis and cryptosporidiosis, however, there is little evidence about their real importance in the maintenance and dissemination of these protozoosis. In order to evaluate the occurrence and determine the genotypes responsible for neotropical mammals infection, 452 fecal samples of 52 different species, in situ and ex situ, of seven locations were evaluated by microscopic methods of diagnosis, followed by molecular amplification (Nested PCR), sequencing and genotypic chacacterization techniques. The results revealed an apparent prevalence of 6,2% for Giardia spp. and 4,8% for Cryptosporidium spp. (n=343). Seventeen different species of wild mammals were positive, 11 for Giardia spp., nine for Cryptosporidium spp. and three for both protozoans. Molecular characterization shows predominant presence of zoonotic genotypes in captive mammals (Giardia duodenalis genotype AI) and host-specific genotypes in free-living animals (Cryptosporidium sp. rat genotype III and Cryptosporidium wrairi). Giardia duodenalis genotype D in crab-eating fox Cerdocyon thous and Cryptosporidium sp. deer mouse genotype IV in black howler monkey Alouatta caraya, both in captivity, were also identified. Ecological aspects, like habitat, trophic guilds, strategy of using the environment and human influence, were considered relevant to occurrence of the parasites. These findings could demonstrate wild mammals can be infected by zoonotic and specific genotypes of agents, which shows the importance of studies using this approach to suggest possible relationships between protozoans, human hosts, domestic animals and wildlife facing different environmental characteristics.

As doenças de veiculação hídrica são uma preocupação no mundo todo, e tem aumentado cada vez mais o interesse nos estudos relacionados aos protozoários Cryptosporidium spp. e Giardia spp.. A presença de protozoários em águas de abastecimento está vinculada ao lançamento de esgoto nos mananciais, de modo que o tratamento de esgotos sanitários é de extrema importância para reduzir o risco de surtos de protozooses. Este trabalho avaliou a ocorrência e a possibilidade de remoção de cistos de Giardia spp. e oocistos de Cryptosporidium spp. em dois reatores UASB, um piloto e um em escala plena, operados com esgoto sanitário e um TDH de aproximadamente 8h, e um reator UASB piloto, operado com água negra simulada e um TDH aproximado de três dias. Nos dois reatores operados com esgoto sanitário as remoções foram semelhantes. A média de DQO total afluente, no reator piloto e no de escala plena, foi de 271 e 410 mg/L, respectivamente, e as remoções médias de DQO foram de 50,62 e 45,80%, respectivamente. Com relação aos exames parasitológicos, quase não houve remoção em ambos os reatores e, na maior parte dos casos, o número de (oo)cistos detectados no efluente foi maior do que no afluente. Já no reator piloto operado com águas negras, foram observadas remoções mais elevadas. A DQO total afluente desse reator variou de 837 a 3269 mg/L e a remoção média observada foi de 71,22%. Neste reator também foram observadas maiores remoções de (oo)cistos, sendo que a remoção de cistos Giardia spp. variou de 67,73 a 93,69% e, em um dos ensaios, foi observada uma remoção de 70% de oocistos de Cryptosporidium spp. Acredita-se que o melhor desempenho desse reator se deve ao seu elevado tempo de detenção hidráulico. Foi avaliada, ainda, a ocorrência de (oo)cistos no lodo dos três reatores, resultando em valores bastante elevados, da ordem de 105 cistos/g de massa seca, em todos eles. De qualquer modo, em todos os reatores avaliados, mesmo o que apresentou as maiores eficiências, o efluente ainda apresentava altas concentrações de poluentes, necessitando de um pós-tratamento para a remoção dos remanescentes de matéria orgânica, sólidos e patógenos. / The waterborne diseases are a concern worldwide, and has grown more and more interest in studies related to the protozoa Cryptosporidium spp. and Giardia spp. The presence of protozoa in drinking water is related to the release of sewage in water sources. So that, the sewage treatment is extremely important to reduce the risk of outbreaks protozoa. This study evaluated the occurrence and the possibility of removal of Giardia spp. and Cryptosporidium spp. in two UASB reactors, a pilot and a full-scale, operated with sewage and a HRT of 8 hours, approximately, and a pilot UASB reactor, operated with simulated black water and an approximate HRT of three days. Removals were similar in both reactors operated with sewage. The average influent total COD, in pilot and full scale reactors, was 271 and 410 mg/L, respectively, and the average COD removal were 50.62 and 45.80%, respectively. With the parasitological test, there was almost no removal in both reactors and, in most cases, the number of (oo)cysts detected in the effluent was higher than in the influent. In the pilot reactor operated with black water, higher removals were observed. The total COD influent of this reactor ranged from 837 to 3269 mg/L and the average removal observed was 71.22%. In this reactor (oo)cysts removals were also higher, and the removal of Giardia cysts ranged from 67.73 to 93.69%, and in one of the trials, a 70% removal of Cryptosporidium spp was observed. It is believed that the best performance of this reactor is due to its high hydraulic retention time. The occurrence of (oo)cysts in the sludge of the three reactors was also evaluated, resulting in very high values of the order of 105 cysts/g of dry weight, in all of them. Anyway, in all this reactor, even the one with the highest efficiency, the effluent still had high concentrations of pollutants, necessitating a post treatment for the removal of remaining organic matter, solids and pathogens.

Faces provide humans with information on the age, sex, individual identity, and emotional state of others. Although comparatively less is known about nonhuman primates’ face processing abilities, several gregarious group living species are able to discriminate conspecific faces. Here, we tested a less gregarious species, orangutans, to determine if they exhibit similar skills. Using a matching-to-sample paradigm, orangutans matched two identical portraits of unfamiliar orangutans. Next, subjects matched two different photographs of the same individual across viewpoints. During testing, subjects successfully transferred to novel photographs of familiar, but not unfamiliar, individuals with their first exposure to these stimuli. However, performance was not maintained throughout continued exposure to these stimuli, suggesting a possible novelty effect. Interestingly, orangutans performed significantly above chance when individuating familiar males, but not females. Further examination is needed to understand social organization and other social factors which were important in the evolution of face-processing.

The effects of selected antimicrobials, applied singularly or in combination, and frozen or refrigerated storage conditions on the survival of Listeria spp. on inoculated shrimp was evaluated in this study. A combination of 0.5% CPC (Cetylpyridinium Chloride) with a water wash at room temperature and freezing of the shrimp at -22.3 degrees C was the only treatment that had a significant antimicrobial effect on the Listeria spp. Antimicrobial effects and the mode of action of PEF (Pulsed Electric Field) and CPC on Listeria cells were evaluated in detailed studies. PEF in 0.1% sodium chloride had a bacterostatic effect toward Listeria spp. during refrigerated storage, but no immediate or bacteriostatic effect was caused by freezing the samples. A concentration of 1% sodium chloride reduced the Listeria spp. population after freezing by 1.1 log; however, the pungent chlorine odor that was generated during treatment might cause discomfort for employees in shrimp processing facilities. Also, chlorine might cause corrosion of metal surfaces of processing equipment. There was no difference in the antimicrobial effects on the survival of Listeria spp. by PEF between the exposure times of 1 or 2 min, as well as in the sodium chloride concentrations of 0.1 and 0.5%. PEF treatment in the presence of 0.1% sodium chloride is recommended. A solution of 0.5% CPC effectively inhibited all of the strains of Listeria spp. in the cell suspensions. A treatment of 0.5% CPC combined with PEF treatment in a sodium chloride concentration of 0.1% caused a delayed effect on the Listeria spp. after 2 d of refrigerated storage. After 2 d of frozen storage, the formation of ice crystals was decreased in the number of Listeria spp. when contaminated samples were treated with water. The results indicated chemicals (e.g. CPC and NaCl) might protect Listeria spp. from the formation of ice crystals. TEM (Transmission Electron Microscopy) micrographs revealed that cell membranes were damaged by PEF treatment and that cells were ruptured by CPC treatment. A maximum reduction of 2.76 log10 CFU/g of Listeria spp. on shrimp was achieved by a combination of PEFCPC.

Faces provide humans with information on the age, sex, individual identity, and emotional state of others. Although comparatively less is known about nonhuman primates’ face processing abilities, several gregarious group living species are able to discriminate conspecific faces. Here, we tested a less gregarious species, orangutans, to determine if they exhibit similar skills. Using a matching-to-sample paradigm, orangutans matched two identical portraits of unfamiliar orangutans. Next, subjects matched two different photographs of the same individual across viewpoints. During testing, subjects successfully transferred to novel photographs of familiar, but not unfamiliar, individuals with their first exposure to these stimuli. However, performance was not maintained throughout continued exposure to these stimuli, suggesting a possible novelty effect. Interestingly, orangutans performed significantly above chance when individuating familiar males, but not females. Further examination is needed to understand social organization and other social factors which were important in the evolution of face-processing.

The alkaloid content of three Crinum species namely C. bulbispermum, C. macowanii and C. moorei was investigated. The ethanolic extracts of C. bulbispermum yielded seven compounds. The new alkaloids 8α-ethoxyprecriwelline, N-desmethyl-8α-ethoxypretazettine and N-desmethyl-8β-ethoxypretazettine were isolated for the first time from a natural source. In addition, the known alkaloids bulbispermine, crinamine, 6-hydroxycrinamine and 3-O-acetylhamayne were isolated in this study. The ethanolic extracts of C. moorei were found to contain Iycorine, 1-O-acetyllycorine, crinine, 3-O-acetyllycrinine, epibuphanisine, powelline, crinamidine, undulatine, epivittatine, 1-epideacetylbowdensine, cherylline and the new alkaloids mooreine and 3-[4'-(2'-aminoethyl)phenoxy]bulbispermine. The alkaloids crinine, lycorine, bulbispermine, cherylline and hamayne were obtained from the ethanolic extracts of C. macowanii. In addition, the amine tyramine was identified during the isolation process. Dilute HCl solution extraction followed by GC analysis was used to investigate organ-to-organ and seasonal variation of alkaloids in each Crinum species, as well as the interspecific variation in these alkaloids over two consecutive years. Twelve alkaloids were identified, including crinine, epibuphanisine, powelline, crinamine, crinamidine, 6-hydroxycrinamine, 1-epideacetylbowdensine, 3-O-acetylhamayne, undulatine, Iycorine, 1-O-acetyllycorine and cherylline. Alkaloids were detected in all organs of C. moorei and C. macowanii. However, alkaloids were not detected in the leaves of C. bulbispermum. Organ-to-organ and seasonal variations in the total yield and total ring types of these alkaloids were noticed. Organ-to-organ and seasonal statistical variations were also detected for some of the individual alkaloids detected in each of these species. The results also showed that C. moorei had the highest levels of all individual alkaloids except crinamine when compared to C. bulbispermum and C. macowanii. Quantitatively, the detected alkaloids chemotaxonomically separated C. moorei from C. bulbispermum and C. macowanii. The results also indicated that C. macowanii is more closely related to C. bulbispermum. Qualitatively, Iycorine, 1-O-acetyllycorine, cherylline, crinamidine, 1-epideacetylbowdensine, crinine, crinamine and 3-O-acetylhamayne were detected in both C. moorei and C. macowanii, indicating the close relationship of these species. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2000.

Like other aromatic hydrocarbons, benzene is a common soil and groundwater contaminant. It is recognized as a human carcinogen. Exposure of benzene can cause serious negative impacts on human health. Benzene is of major concern due to its toxicity and relatively high water solubility. Benzene is easily biodegraded by ubiquitous bacteria with the presence of free oxygen. However, soil and groundwater contamination with petroleum hydrocarbon often results in the development of anaerobic zones. Bioremediation has been considered as an advantageous alternative in terms of fairly low cost, process flexibility, and on-site utility for the treatment of contaminated soil and groundwater. However, benzene is particularly persistent under anaerobic condition even in the enhanced anaerobic biodegradation process. Although studies have shown that benzene biodegradation could occur under several reducing conditions, the in situ activities of anaerobic benzene degradation are generally low. Bioaugmentation rather than biostimulation may be applicable to accelerate biodegradation process. Successful bioaugmentation requires the inoculation of contaminated soil and groundwater with the strains or consortia of specific degrading capabilities. However, information of dominant species within the microorganisms for anaerobic benzene degradation is still limited. To address this problem, in this study, a benzene-degrading nitrate-reducing culture was established with soil contaminated by gasoline. A nitrate-reducing medium with sulphate, phosphate and other inorganic nutrient was employed to enhance anaerobic benzene degradation. BioSep BioTrap coupled with stable isotope probing and other molecular biological methods were used to identify key anaerobic benzene degraders. Members of genus Dokdonella spp., Pusillimonas spp., and Advenella spp. were found to be the dominant microorganisms during anaerobic benzene degradation, and were hypothesized to be benzene degrader under nitrate-reducing condition.