Academic literature on the topic 'Coprostanol'

Selected sterols (coprostanol, epicoprostanol, cholesterol, cholestanol), stanone (5ßcoprostanone) and linear alkylbenzenes (LABs) were measured in the surface sediments near Ferraz station sewage outfalls, in Admiralty Bay, King George Island, South Shetland Islands. during the summer of 1997/98 using GC-FID and GC-MS. Total sterol concentrations varied between 0.21 and 10.4 μg g−1 dry sediment. Cholesterol was the major sterol at all sites, except at the sewage outfall, where coprostanol predominated. The concentration of coprostanol varied between 0.03 and 6.14 μg g−1 dry sediment, but the majority of the samples contained levels below 0.13 μg g−1 dry sediment. The parameters coprostanol+epicoprostanol in total sterols, coprostanol/epicoprostanol ratio versus % of cholesterol in total sterols and versus 5ß-coprostanone concentration were used to identify the sewage impacted locations in the study area. Only sites extending to 50m from the sewage outfall exhibited a sterol signal indicating sewage input. Total LABs varied from <0.60 to 11.8 ng.g−1 dry sediment with the maximum level at the sewage outfall. Faeces from different species of seals all contained large amounts of cholesterol and some 5ßcoprostanone. Relatively low levels of coprostanol and high levels of cholesterol observed in distant sites could be attributed to natural sources such as marine mammals.

Abstract. Sterol ratios are used to identify sources, occurrence and partitioning of faecal matter in sediments of the Tay Estuary, Scotland. The 5β/(5α+5β) ratio is used to discriminate between sewage and biogenic sterol sources by comparing the concentrations of coprostanols to cholesterol plus coprostanols. This index shows unambiguous sewage pollution in the Invergowrie Bay area (values >0.7). The coprostanol/epicoprostanol index is used to differentiate between human and non-human faecal inputs. Ratios confirmed the primary source as human-derived faecal material. The coprostanol/cholesterol ratio was calculated in order to elucidate the contribution of different biogenic sources to the sedimentary sterol budget. Ratios of >1 clearly indicate faecal sterol sources. Invergowrie Bay displayed no sterol signature other than sewage. A biogenic source of cholesterol influenced total sterol concentrations upstream of the City of Dundee. Attention is directed to the potential role of density fronts in compartmentalization of faecal material in bottom sediments.

Every day, up to 1 g of cholesterol, composed of the unabsorbed dietary cholesterol, the biliary cholesterol secretion, and cholesterol of cells sloughed from the intestinal epithelium, enters the colon. All cholesterol arriving in the large intestine can be metabolized by the colonic bacteria. Cholesterol is mainly converted into coprostanol, a non-absorbable sterol that is excreted in the feces. Interestingly, cholesterol-to-coprostanol conversion in human populations is variable, with a majority of high converters and a minority of low or inefficient converters. Two major pathways have been proposed, one involving the direct stereospecific reduction of the Δ5 double bond direct while the indirect pathway involves the intermediate formation of 4-cholelesten-3-one and coprostanone. Despite the fact that intestinal cholesterol conversion was discovered more than a century ago, only a few cholesterol-to-coprostanol-converting bacterial strains have been isolated and characterized. Moreover, the responsible genes were mainly unknown until recently. Interestingly, cholesterol-to-coprostanol conversion is highly regulated by the diet. Finally, this gut bacterial metabolism has been linked to health and disease, and recent evidence suggests it could contribute to lower blood cholesterol and cardiovascular risks.

ABSTRACT The microbial community in the human colon contains bacteria that reduce cholesterol to coprostanol, but the species responsible for this conversion are still unknown. We describe here the first isolation and characterization of a cholesterol-reducing bacterium of human intestinal origin. Strain D8 was isolated from a 10−8 dilution of a fresh stool sample provided by a senior male volunteer with a high capacity to reduce luminal cholesterol to coprostanol. Cholesterol-to-coprostanol conversion by strain D8 started on the third day, while cells were in stationary phase, and was almost complete after 7 days. Intermediate products (4-cholesten-3-one and coprostanone) were occasionally observed, suggesting an indirect pathway for cholesterol-to-coprostanol conversion. Resting-cell assays showed that strain D8 could reduce 1.5 μmol of cholesterol/mg bacterial protein/h. Strain D8 was a gram-negative, non-spore-forming, rod-shaped organism identified as a member of the genus Bacteroides closely related to Bacteroides vulgatus, based on its morphological and biochemical characteristics. The 16S rRNA gene sequence of strain D8 was most similar (>99.5%) to those of two isolates of the recently described species Bacteroides dorei. Phylogenetic tree construction confirmed that Bacteroides sp. strain D8 clustered within an independent clade together with these B. dorei strains. Nevertheless, no cholesterol-reducing activity could be detected in cultures of the B. dorei type strain. Based on Bacteroides group-specific PCR-temporal temperature gradient gel electrophoresis, there was no correlation between the presence of a band comigrating with the band of Bacteroides sp. strain D8 and cholesterol conversion in 11 human fecal samples, indicating that this strain is unlikely to be mainly responsible for cholesterol conversion in the human population.

Abstract Coprostanol is a major fecal sterol in humans and may, therefore, be a good indicator of sewage-polluted waters. Some types of edible seafood, such as clams, that live in these waters may be contaminated with coprostanol. Coprostanol from clam tissue extracts had been previously quantitated by gas chromatography (GC). In the present work, capillary column GC was used to separate coprostanol, and electron ionization mass spectrometry was used to confirm its identity. Confirmation of identity of coprostanol at the 75 ng level was obtained by comparing the spectrum of the authentic standard with spectra of the clam tissue extract obtained under the same instrumental conditions. Various other compounds can be eliminated as potential interferences by virtue of either their different GC retention times or their spectra.

The gut microbiota plays a key role in cholesterol metabolism, mainly through the reduction of cholesterol to coprostanol. The latter sterol exhibits distinct physicochemical properties linked to its limited absorption in the gut. Few bacteria were reported to reduce cholesterol into coprostanol. Three microbial pathways of coprostanol production were described based on the analysis of reaction intermediates. However, these metabolic pathways and their associated genes remain poorly studied. In this review, we shed light on the microbial metabolic pathways related to coprostanol synthesis. Moreover, we highlight current strategies and future directions to better characterize these microbial enzymes and pathways.

In September 2002, nine sediment samples (0-2 cm) were collected from Botafogo Cove (southwestern part of Guanabara Bay) in order to compare the use of chemical (coprostanol) and biological markers (E. coli and total coliforms) in identifying faecal contamination. The values found (organic carbon - 6.0 to 64.8 mg g-1; coprostanol - 1.4 to 105 µg g-1; E. coli - < 30 to 2400 NMP/10g and total coliforms - 40 to 9300 NMP/10g) were similar to or even higher than those observed in other contaminated areas of Guanabara Bay. In stations close to the shoreline, both coprostanol concentration and bacteria count confirmed that domestic sewage had accumulated in the cove. Coprostanol concentrations were still relatively high in the stations furthest from contamination sources, although faecal material represented a smaller fraction of the total organic carbon originated by dilution in waters containing algae. Reduction was proportionately higher by virtue of the use of the colimetric assessment method. This result may be associated with the effect that environmental variables (light, salinity, temperature) may have on the survival of bacteria during transport and deposition of sewage particles. Consequently, based on these results, coprostanol may be considered the better indicator of the faecal contamination level in Botafogo Cove sediments.

Sterols and triterpenols present in sedimentary cores from 12 stations along the Cananéia-Iguape estuarine-lagoonal system were investigated by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Ten sterols and three triterpenols were identified and quantified, indicating both natural and anthropogenic sources. The relative distributions of sterol and triterpenol showed that the study area is submitted to organic matter (OM) from the Ribeira de Iguape River, seawater, surrounding vegetation, and plankton production. The contribution of these sources depends on the region of the estuarine-lagoonal system and the depth of sediment. Regarding anthropogenic sources, only the samples submitted to freshwater flow from the Ribeira de Iguape River presented concentration of coprostanol higher than the threshold value and diagnostic ratios, coprostanol/(coprostanol + cholestanol) and coprostanol/cholesterol, that indicate moderate contamination by domestic sewage in that area of the estuarine-lagoonal system. Therefore, the approach used herein identified the OM sources and its transport along the Cananéia-Iguape estuarine-lagoonal system (Brazil), which is a complex of lagoonal channels located in a United Nations Educational, Scientific and Cultural Organization (UNESCO) Biosphere Reserve.

Abstract Existing methods for the identification of mammalian fecal particles in foods have not heen completely satisfactory because visual identification of small particles is difficult. In addition, identification of feces by determining the presence of fecal alkaline phosphatase is limited to specimens in which the enzyme has not been inactivated, and it does not work well with feces from herbivores. A new method has been developed which uses coprostanol as a fecal indicator. Coprostanol is a heat-stable sterol found in the feces of mammals and some birds. A hexane extract of the suspect particle is applied to the preadsorbent zone of a silica gel thin layer chromatography plate which has been impregnated with 5% phosphomolybdic acid in ethanol. The plate is developed in diethyl ether-heptane (55 + 45), heated, and examined visually for the presence of coprostanol. Amounts of rat feces as small as 0.15 mg and cow feces as small as 0.5 mg have been identified using this method.

Abstract Mammalian feces contain coprostanol (5β-cholestan-3/J-ol). In this study, 7 collaborators each tested 45 unknown specimens by a thin layer chromatographic method that uses coprostanol as an indicator of feces. The materials tested were 5 replicates each of 3 test portion sizes (0.5,1.0, and 5.0 mg) of cockroach excreta (negative), and cow and rat feces (both positive). Of 315 specimens tested, 261 (82.9%) were correctly identified; there were 5 false positives, 26 false negatives, and from 1 collaborator, 23 inconclusive results.

Pollution from human and animal faecal waste is a major cause of deteriorating water quality and increased nutrient loads in coastal and inland waterways. Management of this problem depends on knowing which sources of faecal matter are the cause and what is the degree and extent of the pollution. Bacterial indicator organisms have long been the principal method used to test water samples for faecal contamination. However, none of the currently used bacterial indicators on their own are source specific enough to distinguish different sources of faecal matter. The use of faecal sterol biomarkers in conjunction with existing bacterial indicators offers a new way to distinguish sources of faecal contamination. This study investigates the sources of faecal sterols, the relationship of coprostanol to existing bacterial indicators of faecal pollution, the degradation of faecal sterols and the problem of determining the sources of faecal contamination and the distribution of faecal contamination using faecal sterol biomarkers. 5p-Stanols (i.e. faecal sterols) were found to be significant constituents of human, herbivore (i.e. cows, sheep etc.) and pig and cat faeces. Human faeces contained 73 ± 4% coprostanol in relation to the sum of coprostanol and 24-ethylcoprostanol and primary treated effluent contained 86 ± 0.4% coprostanol. Herbivore faeces contained 38 ± 4% coprostanol and 62 ± 4% 24-ethylcoprostanol whereas pig faeces contained 50 � 5% of each compound. Both birds and dogs faeces contained either trace amounts of 5B-stanols or they could not be detected. Notable differences were observed in the abundance of Closthdium perfringens spores between the faeces of birds and domestic pets such as cats and dogs. The above differences were subsequently exploited to distinguish faecal contamination in Lake Tuggerah. An examination of the relationships between coprostanol and bacterial indicator concentrations from several environments revealed that 60 and 400 ng L of coprostanol corresponded to currently defined primary and secondary contact limits for bacteria measured as either thermotolerant coliforms or enterococci in the environment. Four degradation experiments showed faecal sterols and related sterols such as cholesterol decay at similar rates. An induction period was observed in all experiments which meant that simple exponential equations to describe the rate of decay of coprostanol were inadequate; a complimentary log - log transformation of the data was used and the equation: Y = l-Exp(-Exp(time x -0.01 + temp x -0.158 + 3.33)) x 100 was derived where Y equals the predicted percentage of coprostanol remaining over time at a given temperature. In terms of persistence in the environment, Clostridium perfringens spores > coprostanol > enterococci > thermotolerant coliforms. Two field studies were undertaken to highlight the use of faecal sterols. In the Lake Tuggerah study, the results indicated that faecal contamination of receiving waters in the Tuggerah Lakes during rain events was significant, but was not derived from human faecal matter; rather it appears to be principally derived from native birds and, to a lesser extent, domestic pets. In the Derwent Estuary study, based on the distribution of the faecal biomarker coprostanol, the mid estuary and parts of the upper estuary (from Newtown Bay to Taroona), were found to be severely contaminated by sewage. In summary, the use of faecal sterols to trace faecal contamination were found to be an invaluable addition to the tools water managers use to investigate faecal pollution.

La réduction du taux de cholestérol (CH) sanguin est un point clé dans la lutte contre les maladies cardiovasculaires. L’efficacité contrastée des médicaments disponibles actuellement ainsi que l’intérêt porté autour du microbiote intestinal dans la régulation de la physiologie de l’hôte nous amènent à envisager cette voie comme alternative thérapeutique. La production de coprostanol (CO), dérivé très peu absorbé du CH, par des bactéries de ce microbiote a été corrélée positivement à une faible cholestérolémie. Les objectifs de cette thèse sont i) d’isoler et d’identifier de nouvelles souches bactériennes ayant cette activité, ii) d’identifier les gènes bactériens responsables de cette transformation et iii) de détereminer l’impact de ce métabolisme sur la physiologie de l’hôte. Nous avons isolé 22 nouvelles souches productrices de CO à partir des selles d’un individu en produisant beaucoup. Nous avons choisi les souches Bacteroides sp. D8 et Bacteroides sp. BV pour la construction de deux banques génomiques et huit autres pour des essais d’implantation in vivo dans le tractus gastro-intestinal (TGI) de souris axéniques. Nous avons identifié 55 clones potentiellement positifs par le criblage fonctionnel des banques génomiques. Leurs analyses supplémentaires devraient nous apporter des informations sur les gènes impliqués dans cette activité. Toutes les bactéries sélectionnées sont capables de coloniser le TGI de la souris axénique. La souche Parabacteroides distasonis est la meilleure souche productrice de CO in vivo. Nous avons testé son effet sur la cholestéolémie chez des souris axéniques soumises à un régime riche en CH sur 11 semaines en comparaison avec une souche non productrice in vitro, B. dorei, et avec des souris conventionnalisées comme contrôle. La souche B. dorei produit du CO in vivo, soulignant l’importance de l’environnement dans l’activité de production de CO déjà supposée d’après la littérature et nos résultats in vitro. Des gènes impliqués dans l’excrétion du CH de l’organisme vers les selles sont surexprimés chez ces souris et celles colonisées avec P. distasonis. Cependant seules ces dernières présentent une cholestérolémie plus faible que les souris conventionnalisées. Le mécanisme impliqué semble indépendant de la production de CO et de l’excrétion de CH car les mêmes quantités de ces composés sont retrouvées dans les selles indépendamment du statut bactérien. Les concentrations en acides biliaires totaux dans la bile et dans les selles sont supérieures pour les souris monocolonisées comparées au conventionnalisées. Les selles des souris colonisées avec P. distasonis présentent plus d’acides urso- et chénodésoxycholiques que les souris conventionnalisées et plus d’acide cholique que les souris colonisées avec B. dorei. En conclusion, nous avons isolé de nouvelles souches et identifier des clones potentiellement positifs. Les études in vivo tendent à montrer que l’activité de production de coprostanol n’a pas d’effet sur la cholestérolémie. En revanche, la souche P. distasonis semble diminuer la cholestérolémie par un mécanisme encore inconnu
Cholesterol (CH) level management is a keystone to limit cardiovascular diseases. The contrasted efficiency of the drugs currently available as well as the interest around the intestinal microbiota in regulating the host physiology lead us to consider this pathway as a therapeutic alternative. The production of coprostanol (CO), a very poorly absorbed CH derivative, by bacteria of this microbiota has been positively correlated with low CH plasma level. The aims of this thesis are (i) isolate and identify new bacterial strains possessing this activity, (ii) identify the bacterial genes responsible for this transformation and (iii) determine the impact of this metabolism on host physiology. We isolated 22 new strains producing CO from the stools of a high-coprostanol producing individual. We chose Bacteroides sp. D8 and Bacteroides sp. BV for the construction of two genomic libraries and eight others for in vivo implantation tests in the gastrointestinal tract (GIT) of germ-free mice. We identified 55 potentially positive clones by functional screening of these genomic libraries. Their additional analyzes should provide us with information about the genes involved in this activity. All selected bacteria are capable of colonizing the GIT of germ-free mice. Parabacteroides distasonis is the best strain producing CO in vivo. We tested its effect on blood cholesterol level in germ-free mice subjected to an 11-week CH-rich diet compared to an in vitro non-producing strain, B. dorei, and with conventionalized mice as control. The B. dorei strain produces CO in vivo, emphasizing the importance of the environment in the CO production activity already assumed from the literature and our results in vitro. Genes involved in the excretion of CH from body to feces are overexpressed in these mice and those colonized with P. distasonis. However, only the latter have lower cholesterolemia than conventional mice. The mechanism involved appears to be independent of CO production and CH excretion because the same amounts of these compounds are found in feces independently of bacterial status. Total biliary acids concentrations in bile and feces are higher for monocolonized mice compared to conventionalized mice. The feces of mice colonized with P. distasonis exhibited more urso- and chenodeoxycholic acids than conventionalized mice and more cholic acid than mice colonized with B. dorei. In conclusion, we have isolated new strains and identified potentially positive clones. In vivo studies tend to show that coprostanol production activity has no effect on plasma cholesterol. In contrast, P. distasonis seems to decrease plasma cholesterol by a still unknown mechanism

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Os esteróis fecais são considerados importantes indicadores de contaminação fecal em ambientes aquáticos. São compostos orgânicos hidrofóbicos e encontram-se associados ao material particulado e podem ser preservados por um longo tempo com biodegradação pouco significativa. As relações entre o coprostanol e os demais esteróis encontrados no meio ambiente são importantes para confirmar a presença de esgoto doméstico em ambientes aquáticos. O presente trabalho desenvolveu uma metodologia analítica para extração, separação e determinação dos esteróis coprostanol, epicoprostanol, colesterol, colestanol e sitosterol em amostras de sedimento. Em seguida, aplicou-se o método em amostras de sedimentos coletadas em 6 córregos da cidade de Juiz de Fora. O método de análise dos 5 esteróis foi otimizado em Cromatografia Líquida de Alta Eficiência (HPLC) utilizando coluna cromatográfica de fase reversa octadecilsilano (C-18) com detecção na região do ultravioleta. Para otimizar o processo de separação dos esteróis por HPLC, a composição e o fluxo da fase móvel por eluição isocrática foram otimizados, assim como o comprimento de onda de detecção. Como os esteróis não possuem absortividade molar nessa região do espectro, houve a necessidade de realizar o processo de derivatização. Para isso, foram testados dois reagentes derivatizantes para aumentar a sensibilidade do método. Desse modo, pode-se separar e quantificar os 5 esteróis fecais. Ao concluir essa etapa, aplicou-se essa metodologia para os estudos de tratamento de amostra. O resultado desse estudo permitiu percentuais adequados de recuperação dos analitos. Após o tratamento da amostra, realizou-se o procedimento de derivatização otimizado dos extratos para posterior análise por HPLC. Os níveis de concentração de coprostanol nas amostras de sedimento analisadas variaram entre 0,0050 e 14,48 µg.g-1. Com exceção da amostra do córrego Fazenda Floresta, todas as outras foram consideradas contaminadas por fezes utilizando os parâmetros e critérios de avaliação para esse tipo de contaminação.
The fecal sterols are considered important indicators of fecal contamination in aquatic environments. Organic compounds are hydrophobic and are associated with the particulate material and can be preserved for a long time with negligible degradation. The relationships between coprostanol and other sterols found in the environment are important to confirm the presence of domestic sewage into aquatic environments. The present work developed an analytical method for extraction, separation and determination of sterols coprostanol, epicoprostanol, cholesterol, cholestanol and sitosterol in sediment samples. Then applied the method in sediment samples collected from six streams in the city of Juiz de Fora. The method of analysis of the five sterols was optimized (High Performance Liquid Chromatography (HPLC) using reverse phase chromatography column octadecylsilane (C-18) with detection in the ultraviolet region. To optimize the separation of sterols by HPLC, the composition and flow of mobile phase for isocratic elution were optimized, as well as the wavelength of detection. Because sterols are not molar absorptivity in this spectral region, it was necessary to make the process of derivatization. For this, we tested two derivatizing reagents to increase the sensitivity of the method. Thus, it can separate and quantify the five fecal sterols. When you complete this step, we applied this methodology for studies of treatment samples. The result of this study allowed percentage recovery of analytes suitable. After treatment the sample was held optimized the procedure for derivatization of the extracts for analysis by HPLC. The concentrations of coprostanol in the sediment samples analyzed varied between 0.0050 and 14.48 µg.g-1. Except for the sample stream Farm Forest, all others were considered contaminated by feces using the parameters and evaluation criteria for this type of contamination.

Os rios Iguaçu e Barigüi, nos trechos afetados pelo vazamento da REPAR (2000), foram amostrados em 18 pontos, em 3 campanhas (2007-2008). TPH, BTEX e HPA ocorrem na água com concentrações muito baixas ou nem foram detectados. Não há aporte de hidrocarbonetos proveniente da REPAR. No sedimento, não foram detectados BTEX, HPA têm concentrações baixas ou não foram detectados; TPH ocorre em concentrações 10 vezes menores que o padrão ambiental, mas foi detectado em todas as amostras. As maiores concentrações ocorrem no rio Iguaçu, a jusante da foz do Barigüi. Alguns HPA presentes no óleo derramado só ocorrem neste trecho, sugerindo origem a partir do rio Barigüi. As razões diagnósticas de HPA no sedimento não estabelecem ligação direta com o vazamento da REPAR, por predominarem indicações de fonte pirolítica ou mista. Entretanto, estudo do sedimento do Banhado 4 mostrou que muitas das razões diagnósticas de HPA que indicavam fonte petrogênica no óleo original passaram a indicar fonte pirolítica 7 anos após o acidente. O índice pirogênico é a única razão que manteve indicação de fonte petrogênica. Na estação seca, as concentrações médias de esteróis no sedimento do rio Barigüi são coprostanol (25,97 μg/g), coprostanona (4,67 μg/g), colesterol (9,02 μg/g), colestanol (4,15 μg/g) e colestanona (0,42 μg/g). Na estação chuvosa, as concentrações reduzem-se pela metade (colestanona não foi detectada). O rio Iguaçu apresenta resultados similares a montante de Fazenda Rio Grande. Mas, nesta localidade, as concentrações aumentam apenas na estação chuvosa para coprostanol (249,63 μg/g), coprostanona (11,6 μg/g), colesterol (81,85 μg/g) e colestanol (49,99 μg/g), o que é atribuído à lavagem do solo pela água pluvial. A influência de Araucária é marcada por altas concentrações de coprostanol (329,8 μg/g na estação seca e 229,25 μg/g na estação chuvosa). Daí até General Lúcio as maiores concentrações ocorrem no canal principal, na campanha seca (média de 246,08 μg/g); na campanha chuvosa, a média cai para 101,51 μg/g. Nos canais secundários, o comportamento é contrário, relacionado ao acúmulo de sedimentos contaminados ocorrer aí apenas na cheia. No rio Iguaçu, os valores médios de COT na estação chuvosa e na seca são semelhantes (~28 mg/g) e bem maiores do que no rio Barigüi (estação chuvosa 5,2 μg/g; seca 8,9 mg/g). As relações entre coprostanol e COT indicam que o aporte de esgoto é o principal fator de definição do teor de carbono orgânico sedimentar apenas para o canal principal do rio Iguaçu e na estação chuvosa. Nos canais secundários deste rio e no rio Barigüi, em qualquer estação, ocorre contribuição importante de outras fontes (vegetação e/ou produção primária in situ). Coprostanol e coprostanona apresentam forte correlação nas duas estações. A ausência de correlação entre coprostanol e colesterol na estação seca, nos dois rios, pode ser explicada pela degradação do colesterol e/ou por parte deste provir de fonte diferente do coprostanol. As razões [5β/(5α+5β)estanona] e [5β/(5α+5β)estanol] são superiores a 7 (altamente contaminado) em 16 pontos na estação chuvosa e em todos os 18 pontos na estação seca.
The Iguaçu and Barigui rivers in parts affected by leakage REPAR (2000), were sampled in 18 points in three campaigns (2007-2008). TPH, BTEX and PAH occurring in water with very low concentrations or not detected. There is no input of oil from the REPAR. In the sediment, were not detected BTEX, PAH concentrations are low or not detected; TPH occurs in concentrations 10 times lower than the environmental standard, but was detected in all samples. The highest concentrations occur in the Iguaçu river, downstream from the mouth of Barigui. Some PAHs present in the oil spill occur only in this stretch, suggesting origin from the river Barigüi. The reasons diagnostic of PAH in the sediment do not establish a direct connection with the leaking of REPAR, predominate source of indication by pyrolytic or mixed. However, a study of sediment from the Wetland 4 showed that many of the reasons diagnostic for PAH that indicated petrogenic source in the original oil passed indicate source pyrolytic seven years after the accident. Pyrogenic index is the only reason that kept indication of petrogenic source. In the dry season, the average concentrations of sterols in the sediment of Barigüi river are coprostanol (25.97 μg/g), coprostanona (4.67 μg/g), cholesterol (9.02 μg/g), cholestanol (4.15 μg/g) and Cholestanones (0.42 μg/g). In the rainy season, the concentrations are reduced by half (Cholestanones was not detected). The Iguaçu river shows similar results upstream of Fazenda Rio Grande. But in this location the concentrations increase only during the rainy season to coprostanol (249.63 μg/g), coprostanona (11.6 μg/g), cholesterol (81.85 μg/g) and cholestanol (49.99 μg/g), which is assigned to wash the soil by rainwater. The influence of Araucaria is marked by high concentrations of coprostanol (329.8 μg/g dry season and 229.25 μg/g in the rainy season). Thence to General Lúcio the highest concentrations occur in the main channel in the dry campaign (average of 246.08 μg/g) in the rainy campaign, the average falls to 101.51 μg/g. In the secondary channels, the behavior is opposite, related to the accumulation of contaminated sediments occur there only in full. In the Iguaçu River, the average values of TOC in the rainy and dry seasons are similar (~ 28 mg/g) and much larger than the Barigüi river (wet season 5.2 μg/g dry 8.9 mg/g). The relationships between coprostanol and TOC indicate that the sewage discharges are the main factor defining the sedimentary organic carbon content just for the main channel of the Iguaçu river and the rainy season. In secondary channels this river and in the river Barigüi, in any season, there is significant contribution from other sources (vegetation and/or primary production in situ). Coprostanol and coprostanona have strong correlation in both seasons. The lack of correlation between cholesterol and coprostanol in the dry season in the two rivers can be explained by the degradation of cholesterol and/or part of this come from different source of coprostanol. The reasons [5β / (5α +5 β) estanona] and [5β / (5α +5 β) stanol] are superior to 7 (highly polluted) on 16 points in the rainy season and all 18 points in the dry season.

In this study we investigated the composition of the sediment from Sergipe River Estuary, located in the metropolitan area of Aracaju, in two periods: the winter of 2010 and the summer of 2011. In the recent years, this river has been undergoing a process of degradation due to release of sewage, without proper treatment, and the process of industrialization in its basin. We performed the sediments characterization from the analysis of total organic carbon (TOC), total organic matter (TOM), total nitrogen (TN) and the amount of silt, clay and sand. The lipid compounds, such as sterols and linear alcohols, were identified and quantified in extracts of surface sediments in the Sergipe River Estuary by gas chromatography coupled to mass spectrometry (GC-MS). The autochthonous and allochthonous contributions were characterized in the study area through the determination of sterols and alcohols in sediments. The autochthonous contribution was determined using the amount and distribution of the linear alcohols of short chain ( C20) and the identification of sterols of 29 carbon atoms. The C30 alcohol was the compound which presented the higher concentration within all samples, with values ranging from 0.62 to 2.46 μg.g-1 in the winter and 0.37 to 21.41 μg.g-1 in the summer. The most prominent sterol concentration was observed for β-sitosterol (C29), in concentrations ranging from 1.15 to 15.69 μg.g-1 in the winter and from 1.23 to 8.09 μg.g-1 in the summer. These compounds are frequently found in large quantities in vascular plants. Therefore, suggesting a strong contribution of terrigenous materials to the estuary. Contamination by sewage in the Sergipe River estuarine system was assessed by the presence of coprostanol (fecal biomarker) in the sediments and the different sterols ratios and indexes. This contamination was attributed to the possible discharge of untreated sewage into the Sergipe river basin.
Neste trabalho foi feita a análise do sedimento do Estuário do Rio Sergipe, situado na região metropolitana de Aracaju, em dois períodos: no inverno de 2010 e no verão de 2011. Nos últimos anos, esse Rio vem passando por um processo de degradação devido ao lançamento de esgotos domésticos, sem tratamento apropriado, em suas águas e pela concentração industrial nessa Bacia. Foi realizada a caracterização dos sedimentos a parir da análise de teores de carbono orgânico total (COT), matéria orgânica total (MOT), nitrogênio total (NT) e teores de silte, argila e areia. Os compostos lipídicos, esteróis e álcoois lineares, foram identificados e quantificados nos extratos de sedimentos superficiais no Estuário do Rio Sergipe através da cromatografia em fase gasosa acoplado a espectrômetro de massas (CG-EM). As contribuições autóctone e alóctone foram caracterizadas na área de estudo através da determinação dos esteróis e álcoois nos sedimentos. A contribuição autóctone foi determinada a partir dos álcoois lineares de cadeia curta (C20) e pela identificação de esteróis com 29 átomos de carbono na estrutura. O álcool identificado em maior concentração em todas as amostras foi o C30, em concentrações que variaram de 0,62 a 2,46 μg.g-1 no inverno e de 0,37 a 21,41 μg.g-1 no verão. O esterol de concentração mais proeminente foi o β-sitosterol (C29) em concentrações que variaram de 1,15 a 15,69 μg.g-1 no inverno e de 1,23 a 8,09 μg.g-1 no verão. Esses compostos são frequentemente encontrados em grandes quantidades em plantas superiores, sugerindo assim uma forte contribuição de material terrígeno no estuário. A contaminação por esgoto no estuário do Rio Sergipe foi avaliada pela presença do coprostanol (biomarcador de origem fecal) nos sedimentos analisados e pelas razões de diferentes esteróis. Esta contaminação foi atribuída ao possível lançamento de esgoto não tratado na Bacia do Rio Sergipe.

Koffein (1,3,7-Trimethylxanthin) und Coprostanol (5beta-cholestan-3beta-ol) wurden im Berliner Oberflächenwasser nachgewiesen. Ihre Konzentrationen korrelierten mit dem Verunreinigungsgrad der Proben, was nahelegt, dass sie sich als Marker für menschliche Aktivität eignen. Bemerkenswerterweise wurde Koffein in jeder einzelnen Oberflächenwasserprobe oberhalb der Bestimmungsgrenze von 0,025 µg/L gefunden. Um Oberflächenwasserproben in größeren Serien zu untersuchen, war die Entwicklung zweier neuer Methoden erforderlich: ein Immunoassay, basierend auf einem monoklonalen Antikörper für Koffein und eine dispersive flüssig-flüssig Mikroextraktionsmethode (DLLME), gefolgt von Flüssigkeitschromatographie gekoppelt mit Tandem-Massenspektrometrie (LC-MS/MS) für Coprostanol. Der entwickelte Koffein-Immunoassay zeigt die beste je erhaltene Nachweisgrenze für Koffein (0,001 µg/L), erlaubt Hochdurchsatz-Analysen und erfordert keine Probenvorbereitung. Der Assay wurde auch erfolgreich für die Messung von Koffein in Getränken, Haarwaschmitteln, Koffeintabletten und menschlichem Speichel angewendet. Antikörper gegen Coprostanol sind nicht kommerziell erhältlich. Eine neue Strategie Anti-Coprostanol-Antikörper zu generieren wurde erarbeitet, die eine analoge Verbindung – Isolithocholsäure (ILA) – als Hapten verwendet, mit der eine Gruppe von Mäusen immunisiert wurde. Ein polyklonales Anti-ILA-Serum wurde produziert, welches Coprostanol bindet, aber die niedrige Affinität erlaubte nicht den Aufbau eines Immunoassays, der die Messung von Umweltkonzentrationen des Anayten (im Bereich ng/L) zulässt. Spezifische Anti-ILA-Immunglobuline G wurden auch in den Faeces der Mäuse gefunden. Coprostanol wurde in den Wasserproben durch die Verwendung einer neuentwickelten LC-MS/MS-Methode unter APCI-Ionisation (atmospheric pressure chemical ionisation) gemessen. Konzentrationen oberhalb von 0,1 µg/L wurden nach Voranreicherung der Probe mittels DLLME bestimmt.
Caffeine (1,3,7-trimethylxanthine) and coprostanol (5beta-cholestan-3beta-ol) were detected in samples of Berlin’s surface water. Their concentrations correlated with the contamination status of the samples, suggesting their usefulness as markers of human activity. Remarkably, caffeine concentrations were always well above the limit of quantitation of 0.025 µg/L. In order to screen surface water samples in larger series, the development of two novel methods was required: a monoclonal antibody-based immunoassay for caffeine and a dispersive liquid-liquid microextraction (DLLME) method, followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) for coprostanol. The caffeine immunoassay developed shows the best analytical limit of detection (LOD) obtained so far for caffeine (0.001 µg/L), allows high-throughput analysis, and does not require sample pre-treatment. The assay was also successfully employed to measure caffeine in beverages, shampoos, caffeine tab-lets, and human saliva. Antibodies to coprostanol are not commercially available. A new strategy to generate anti-coprostanol antibodies was elaborated using an analogous com-pound as hapten – isolithocholic acid (ILA) – and immunizing a group of mice. A polyclonal anti-ILA serum was produced, which binds coprostanol but the low affinity did not permit setting up an immunoassay to measure environmental concentrations of the analyte (in the range of ng/L). Specific anti-ILA immunoglobulin G were also found in the faeces of the immunized mice. Coprostanol was quantified in the water samples using a newly developed LC-MS/MS method using atmospheric pressure chemical ionisation (APCI). Concentrations above 0.1 µg/L were determined after sample preconcentration using DLLME. This extraction method also proved to be successful for enrichment of coprostanol-related compounds such as cholesterol, cholestanol, cholestanone, ergosterol, and stigmasterol.

Faecal indicator bacteria have traditionally been used in the detection of faecal pollution in water, but due to concerns about the lack of reliability of these indicators, alternative methods have been developed. One of which is the detection of sterols present in human and animal excreta via GC-MS analysis of water in this study. The Szűcs method was used to detect six target sterols (coprostanol, cholesterol, dehydrocholesterol, stigmasterol, β-sitosterol, and stigmastanol) in environmental water samples. An initial study was done by analysing raw sewage and effluent (human faecal sterol biomarkers) and water samples were spiked with excreta from cattle, chickens, horses, pigs, and sheep to determine faecal sterol fingerprints. The method was evaluated for quantitation and differences between the water samples from each species. Following liquid-liquid extraction, silylation and derivatization, samples were analysed by GC-MS. Standard curve assays were linear up to 160ng and the limit for quantification was 20ng. The human faecal sterol biomarker was coprostanol, while herbivore profiles were dominated by terrestrial sterol biomarkers (stigmasterol and stigmastanol). Sterol fingerprints and differences in concentrations of sterols between various animals and between animals and humans occurred, providing the opportunity to determine whether faecal pollution was from humans or from animals. The method proved sensitive enough to evaluate faecal contamination in environmental water. Groundwater was collected from bore-holes and surface water samples were collected from the Baberspan Inland Lake. Physico-chemical parameters analysed indicated that pH for surface water samples was above 6.9. The total dissolved solids (TDS) in groundwater indicated that the water was not suitable for human consumption, but could be used for livestock watering. Surface water electrical conductivity (EC) and inorganic nitrates was too high to be used for irrigational purposes. Nitrates in groundwater were too high to be consumed by humans. In groundwater, the total coliform target water quality range (TWQR) was exceeded at 53% of sites analysed and faecal coliform TWQR were exceeded at 77% sites. Surface water samples complied with TWQR with regards to faecal coliforms for full contact recreational activities and livestock watering. The TWQR for E. coli, with regards to full contact recreational activities, was within a safe range for surface water. Faecal streptococci were found in 85% of groundwater sampling sites. And surface water faecal streptococci counts exceeded the TWQR for full contact recreational activities. There is no TWQR for faecal sterols in water, but concentrations of cholesterol and coprostanol was found at three of the groundwater sites analysed. This indicates faecal contamination from possible animal and human origin. Surface water samples analysed showed that the Harts River water is clean and free of faecal sterols, while the water analysed from the inflow, hotel and outflow, cholesterol eluted, which showed faecal contamination, possibly from animals. Faecal sterol markers could be detected in groundwater and surface water, adding an extra dimension to determining the quality of water systems. An optimization and sensitivity study of the method was done on waste water treatment plant (WWTP) raw sewage and effluent. The WWTP sample analysed form Potchefstroom and Carletonville WWTP yielded all six target sterols in the raw sewage water samples, but no sterols eluted in the effluent samples. The raw sewage water sample taken from the Fochville WWTP yielded all six target sterols as well, however, the effluent yielded an unknown compound as well as cholesterol. An alternative study was done where the effluent sample volume was increased. By increasing the volume of water, one can possibly increase the amount (“load”) of sterols extracted and analysed, resulting in a higher abundance of target sterols. By using the target qualifier ions of the six target sterols, and the GC-TOF/MS software, the target sterols could still be qualitatively determined. Optimal volume for raw sewage is 300 ml water sample as this is enough to yield all 6 target sterols. For optimum water quality monitoring via faecal sterol analysis of effluent and other environmental samples, at least 1L sample volume needs to be collected and analysed. The methods described here can be applied to the analysis of environmental water samples. The technical advantages also make it suitable for routine environmental monitoring of faecal pollution.
MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

This chapter examines the application of anthropogenic compounds as biomarkers. Since World War II, human activities have introduced a wide array of compounds into the environment, including insecticides such as dichloro-diphenyl-trichloroethane and pesticides, halocarbons (chlorofluorocarbons), sewage products (coprostanol), and polycyclic aromatic hydrocarbons (PAHs). The chapter introduces structural features of these compounds, their distribution and transformation in the environment, and their potential use(s) as tracers. It presents examples of how relationships between anthropogenic markers and biomarkers can be used to provide information about the sources, delivery, and fate of natural organic matter in aquatic ecosystems. It introduces various emerging contaminants (personal care pharmaceutical products, caffeine, and flame retardants) and their potential use as tracers for anthropogenic organic matter in aquatic ecosystems. It describes how δ‎¹³C, stable isotopes of Cl and Br, and radiocarbon can be used to apportion sources of organic contaminants (e.g., PAHs and PCBs).