Hydrophilic interaction liquid chromatography is a frequently used separation method for analysis of polar compounds. It is an alternative method to reversed-phase chromatography, where these compounds show insufficient or very weak retention. A high number of stationary phases are currently available for HILIC and new ones are still being developed. The aim of this diploma thesis was to characterize and compare three relatively new HILIC columns containing an unmodified silica gel (HILIC-A), aminopropyl modified silica (HILIC-B) and polyhydroxyl chain modified silica (HILIC-N) as stationary phase. Based on the study of the effect of acetonitrile content in mobile phase on the retention of a model set of peptides, a multimodal retention mechanism was demonstrated. Analysis of 18 model analytes with different pKa values showed, that the composition of aqueous part of mobile phase (buffer), more specifically it's concentration and pH value, has a significant impact on retention of ionized analytes and peptides on the studied stationary phases. A significant contribution of ionic interactions to retention was observed on HILIC-B and HILIC-A columns. The retention of basic compounds on the HILIC-B column increased with increasing ionic strength, while it decreased on the HILIC-A column. Increasing the...

Orientador: Roberto de Oliveira Roça / Resumo: Esse estudo objetivou avaliar a eficiência de estímulos elétricos com tensões crescentes durante a estimulação de meias carcaças no final do abate na qualidade do contrafilé até 28 dias de maturação. Metade das meias carcaças esquerdas de 40 novilhos Nelore (Bos taurus indicus), não castrados, com predomínio de cobertura de gordura mediana, foram submetidas ao Protocolo A (20, 40, 60 e 80 VRMS) e a outra metade foi submetida ao Protocolo B (50 VRMS). Ambos os protocolos com 12,5 Hz durante 52 segundos. As respectivas meias carcaças direitas não foram estimuladas (Controle A e Controle B). O pH e a temperatura das meias carcaças foram monitorados durante as 40 horas de resfriamento (1, 3, 6, 9, 12, 24 e 40 horas) e os respectivos contrafilés foram maturados por 2, 7, 14, 21 e 28 dias post-mortem. As duas combinações de estimulação elétrica de média voltagem aceleraram a taxa de declínio do pH (refletido na temperatura e no tempo que o pH cai para 6), resultando em valores médios para força de cisalhamento de 4,66 kgf pra o Protocolo A e 4,62 kg para o Protocolo B com diferença entre seus controles os quais apresentaram 5,36 kg e 5,29 kg respectivamente, mantendo essa característica durante a maturação. O comprimento dos sarcômeros não foi diferente (P>0,05) entre os protocolos e seus controles. O uso da estimulação elétrica com tensões crescentes não resultou em melhores características qualitativas da carne em relação a estimulação elétrica de tensão contínua. A força da esti... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study was aimed to evaluate the efficiency of electrical stimulation with increasing voltage during the stimulation of bovine half carcasses on the quality of 28 days aged strip loins. Half of the left half carcasses from 40 Nellore steers (Bos taurus indicus), intact, from the same feedlot, with median fat coverage, were submitted to Protocol A (20, 40, 60 and 80 VRMS) and the other half to Protocol B (50 VRMS). Both protocols with 12.5 Hz for 52 seconds. The respective right half carcasses were not stimulated (Control A and Control B). The pH and temperature were monitored during 40 hours of cooling (1, 3, 6, 9, 12, 24 and 40 hours) and the strip loins were aged for 2, 7, 14, 21, and 28 days post-mortem. Both combinations of medium voltage electrical stimulation accelerated the pH decline rate (reflected in temperature and the time that pH drop to 6), resulting in lower values for shear force 4.66 kgf for Protocol A, and 4.62 for Protocol B, with 5.36 kgf and 5,29 kgf for its respectively controls, maintaining this characteristic during aging time. The length of the sarcomere was not different (P>0.05) between the protocols and their controls. The use of an electrical stimulation with increasing tensions did not result in better qualitative characteristics of the meat in comparison to the continuous tension electrical stimulation. The strength of the electrical stimulation, the visual contractions, and the pH difference area (∆pH) did not have a proportional effect on ... (Complete abstract click electronic access below) / Mestre

Yes / Fluorescence spectroscopy on a series of aqueous solutions of poly(acrylic acid) containing a luminescent label showed that polymers with molar mass, Mn < 16.5 kDa did not exhibit a pH responsive conformational change, which is typical of higher molar mass poly(acrylic acid). Below this molar mass, polymers remained in an extended conformation, regardless of pH. Above this molar mass, a pH-dependent conformational change was observed. Diffusion-ordered nuclear magnetic resonance spectroscopy confirmed that low molar mass polymers did not undergo a conformational transition, although large molar mass polymers did exhibit pH-dependent diffusion. / Engineering and Physical Sciences Research Council (EPSRC) funded CASE award PhD. Part funded by flocculant manufacturer SNF (UK) Ltd.

[pt] Os medidores de pH existentes no mercado indicam valores confiáveis, dentro de uma incerteza declarada para cada modelo fabricado. Entretanto, normalmente, não é citada a especificação em termos do tempo necessário para alcançar estes valores, devido ao tempo de resposta do instrumento. Esta dissertação de mestrado tem como objetivo desenvolver uma metodologia teórico-experimental para avaliar o desempenho do transdutor em condições transientes, indicando alguns valores típicos para medidores comerciais, e sua resposta na medição de pH, a partir do modelo testado em uma reação experimental, sendo o mesmo simulado em outras reações químicas para diferentes velocidades de reação.Através deste modelo qualificado, foram avaliados os erros devidos às respostas permanentes e transientes, mostrando em que condições o medidor de pH pode ser usado, a partir da comparação entre valores de pH real e valores de pH registrados pelo medidor, levando em consideração a temperatura e o tempo de reação. / [en] The electrochemical analysers available in the market assert trustworthy values applicable in a specific uncertainty range, which varies depending on the unit s model. Nevertheless, rarely is given any information on the time necessary to reach these values, which depend on the answer of the instrument. This Master s dissertation aims to develop an experimental theoretical methodology to evaluate the performance of the transducer in transient conditions, and obtain some typical values for commercial analysers, and its answer in the pH measurements in simulated chemical reactions in various reaction velocities. Through the qualified model, the errors due to the permanent and transient answers were evaluated, establishing the conditions pH electronic units can be used, in a pre-set controlled error, from the comparison between real pH values and displayed pH values in the analysers, considering the temperature and the reaction time.

Les fonctions organiques du corps humain sont liées à des constants biologiques. Variations de ces constantes induisent divers états pathologiques. Parmi ces constantes, le pH constitue le cœur de ces travaux de thèse. Chez les êtres vivants, les fonctions biologiques dépendent de constant acides ou alcalines. En fait, l’action d’une protéine dépend du pH du milieu environnant. Une valeur inadéquate du pH rend les protéines inactives ce qui est délétère pour l’organisme. Il existe donc un besoin pour des capteurs de pH qui puissant être utilisés dans le corps humain pour des applications cliniques (échelle macroscopique), sur des cellules en culture pour des recherches en biologie (échelle mesoscopique) et pour étudier les échanges ioniques au niveau des membranes cellulaires pour des travaux plus fondamentaux (échelle microscopique). Parmi le large éventail de technologies potentiellement candidates pour ces applications, la mesure de pH par fibre optique exploitant la fluorescence permet d’être adaptée aux trois échelles dimensionnelles susnommées. Ce manuscrit de thèse adresse des contraintes par l’étude de capteurs fluorescents à fibre optique utilisant deux types d’indicateur de pH: les SNARF et la fluorescéine. En parallèle de ces développements expérimentaux, des descriptions mathématiques des propriétés de fluorescence de ces deux indicateurs sont proposées. Ces descriptions permettent de progresser vers une mesure du pH sans calibration / Organic functions of the human body are linked to biological constants. Variations of these constants induce pathological troubles. Among these constants, the pH is the central subject of this PhD work. In living beings, biological functions are related to either acid or alkaline constants. Indeed, the action of a protein depends on the surrounding pH. An inadequate value of the pH makes the proteins non active which is deleterious for the organism. There exist a need for pH sensors which can be used in the human body for clinical applications (macroscopic scale), on cells in culture for biology researches (mesoscopic scale) or at a cell membrane level for more fundamental studies (microscopic scale). Among the wide range of technologies potentially useful for these applications, fiber optic fluorescence pH sensing offers the possibility to be adapted to the three above mentioned dimensional scales. This PhD dissertation addresses these constraints by studying fluorescence fiber optics pH sensors using two kind of pH indicators: SNARF and fluorescein. Together with these experimental studies, mathematical descriptions of the fluorescence properties of these indicators are proposed. They allow progressing towards calibration free pH sensing.

Os solos ácidos recobrem, cerca de 40% da área terrestre, constituindo-se em um dos principais fatores limitantes à população à produção vegetal do planeta. No Brasil, os solos ácidos abrangem cerca de dois terços do território nacional. De um modo geral, a acidez do subsolo reduz a profundidade do sistema radicular, aumentando a susceptibilidade à seca e diminuindo o uso de nutrientes pelas plantas. Além da alta atividade de íon hidrogênio (H+), os solos ácidos geralmente apresentam níveis tóxicos de alumínio, sendo que a toxicidade por AI tem sido intensivamente investigada nos últimos anos. No entanto, a toxicidade por AI ocorre apenas a pH baixo e em condições onde a toxicidade por prótons geralmente também se manifesta. Apesar disto, trabalhos envolvendo a toxicidade por prótons são escassos. A sensibilidade de células a pH baixo depende da fase de crescimento e desenvolvimento em que estas se encontram. Em raízes, as células mais sensíveis são as da região de alongamento e em suspensões celulares as células na fase log de crescimento são mais sensíveis do que as células na fase estacionária. Este trabalho faz parte de uma linha de pesquisa que procura explorar as diferenças que existem entre células quanto à sensibilidade a pH baixo para melhor entender a toxicidade e tolerância a prótons. Foram utilizadas células da cultura de tabaco cv. BY-2, um sistema modelo que apresenta diversas vantagens sobre o uso de raízes para a realização deste trabalho. Os objetivos deste estudo foram de a) determinar condições apropriadas para a exposição destas células a acidez; b) caracterizar a resposta destas células a pH baixo, sob influência de diferentes fatores ambientais, quando se encontram em estados distintos de sensibilidade ao pH baixo; e c) verificar se mudanças na sensibilidade das células a pH baixo podem ser decorrentes de alterações na composição da membrana plasmática ou na atividade das ATPases. Vários testes iniciais foram realizados com o intuito de se definir algumas condições experimentais - o tempo de exposição, a composição do meio e o tampão a ser empregado. Optou-se por lavar as células e depois incubá-las por 1h em meio simples com pH desejado e contendo apenas Ca \'Cl IND. 2\' 2mM, KCl 10mM. E o tampão MES ou biftalato (10 mM). O biftalato foi testado porque o tampão MES, usado normalmente no meio de cultura completo, não é eficiente na faixa de pH abaixo de 5,0. O biftalato (pKa = 4,1) praticamente não afetou a viabilidade celular avaliada pela permeabilidade a trypan blue, mas inibiu o crescimento celular no meio de cultura completo. Mesmo assim, os dois tampões foram utilizados em paralelo em diversos experimentos e verificou-se que os resultados foram semelhantes. A viabilidade das células na fase log (2 dias) foi reduzida quando se abaixou o pH de 5,6 a 3,8, sendo que caiu mais acentuadamente até o pH de 4,8. As células da fase estacionaria (7d) foram insensíveis ao baixo pH. A um pH fixo de 4,2 aumentando-se a concentração de Ca \'Cl IND. 2\' para cerca de 8 a 16 mM praticamente aboliu o efeito deletério do pH baixo. Para se ter o mesmo efeito com a adição de KCl, foi necessário adicionar entre 80 e 160 mM. A adição de sacarose também amenizou os efeitos do pH\'sendo praticamente revertido a uma concentração de 100 Mm. Os resultados indicam a importância da força iônica e da osmolaridade da solução, mas o efeito de Ca não parece depender apenas destas duas propriedades. A inibição de ATPases, pelo uso de DCCD, não pareceu ter qualquer relação com a sensibilidade a pH baixo. Tanto células de tabaco na fase log quanto estacionárias foram sensíveis à aplicação de ortovanadato de sódio. Em células da fase estacionária, este efeito mais acentuado a pH 4,2, sugerindo que nestas células, as H+ ATPases do tipo P da membrana plasmática podem ter algum papel na tolerância destas células ao baixo pH. Encontrou-se diferenças no perfil protéico de frações enriquecidas em membrana plasmática entre células da fase log e estacionárias e entre células tratadas ou não a pH baixo. Estas diferenças precisam ser melhor estudadas. / Acid soils account for about 40 % of the surface area of the world and are one of the major factors limiting plant productivity. In Brazil, these soils comprise roughly two-thirds of its total territory. In general, soil acidity is detrimental because it limits the depth of the root system, increasing susceptibility to drought and decreasing the use of nutrients. In addition to the high levels of hydrogen ion activity, acid soils usually have Al toxicity hazards, a topic which has been intensely studied in the past years. However, Al toxicity only occurs at low pH, under conditions in which proton toxicity is also a problem. Despite this, studies of proton toxicity are lacking. The sensitivity of cells to low pH depends on their growth and developmental status. In roots, the most sensitive cells are those of the elongation zone and in cell cultures, cells in the log phase are more sensitive than those of the stationary phase. This study is part of a larger attempt to explore the differences that exist between cells with respect to their sensitivity to low pH to further understand toxicity and tolerance to protons. Cells of tobacco BY-2, a plant cell model system which has several advantages over the use of roots, was used in this study. The objectives were a) to determine the appropriate conditions to expose these cells to low pH; b) characterize the response of these cells to low pH, when different environmental factors are varied and at different stages of cellular sensitivity to acidity; and c) examine if changes in the sensitivity of cells to low pH are due to changes in the composition of plasma membranes or in the activity of ATPases. Several preliminary tests were performed to define the experimental conditions to be used ? the duration of exposure to low pH, the composition of the medium and the buffer to be used. A simple solution containing only CaCl2 2mM, KCl 10 mM and MES or phthalate buffer (10 mM) was chosen to wash and incubate the cells at the pH of interest. Phthalate was tested because MES, the buffer normally used in the culture medium, is not effective at pH values below 5.0. Phthalate (pKa = 4,1) had very little effects on cell viability as evaluated by membrane permeability to trypan blue, but it severely inhibited the growth of the cell culture in complete medium. Nevertheless, both buffers were used in several subsequent experiments, the results of which were found to be similar between both buffers. The viability of log-phase cells (2 day-old) was reduced when the pH was lowered from pH 5,6 to 3.8, but this was sharper down to pH 4,8. Cells in stationary phase (7 day-old) were insensitive to low pH. At a fixed pH of 4,2, increases in the concentration of CaCl2 up to 8 or 16 mM abolished most of the deleterious effects of low pH. To generate the same effect, KCl had to be added at concentrations between 80 and 160 mM. The addition of sucrose also alleviated the effects of low pH. The results suggest the importance of solution ionic strength and osmolarity on sensitivity to low pH, but the effects of Ca do not appear to depend only on these two properties. The inhibition of ATPases, by DCCD, did not appear to bear any relation to cellular sensitivity to low pH. Both log-phase and stationary-phase cells were affected by the addition of sodium orthophosphate. In stationary-phase cells, this effect was more pronounced at pH 4,2, suggesting that in these cells, P-type H+-ATPases of the plasma membrane may play a role in the tolerance of these cells to lo pH. Differences were found in the protein profile of enriched plasma membrane fractions both between log-phase and stationary-phase cells and between cells treated or not with low pH. These differences, however, need to be better examined.

In intense laser – cluster interaction, the experimental results show a strong energetic coupling between radiation and matter. We have measured absolute x – ray yields and charge state distributions under well control conditions as a function of physical parameters governing the interaction; namely laser intensity, pulse duration, wavelength or polarization state of the laser light, the size and the species of the clusters (Ar, Kr, Xe). We have highlighted, for the first time, an intensity threshold in the x – ray production very low (~ 2.1014 W/cm2 for a pulse duration of 300 fs) which can results from an effect of the dynamical polarisation of clusters in an intense electric field. A weak dependence with the wavelength (400 nm / 800 nm) on the absolute x – ray yields has been found. Moreover, we have observed a saturation of the x – ray emission probability below a critical cluster size.

Os solos ácidos recobrem, cerca de 40% da área terrestre, constituindo-se em um dos principais fatores limitantes à população à produção vegetal do planeta. No Brasil, os solos ácidos abrangem cerca de dois terços do território nacional. De um modo geral, a acidez do subsolo reduz a profundidade do sistema radicular, aumentando a susceptibilidade à seca e diminuindo o uso de nutrientes pelas plantas. Além da alta atividade de íon hidrogênio (H+), os solos ácidos geralmente apresentam níveis tóxicos de alumínio, sendo que a toxicidade por AI tem sido intensivamente investigada nos últimos anos. No entanto, a toxicidade por AI ocorre apenas a pH baixo e em condições onde a toxicidade por prótons geralmente também se manifesta. Apesar disto, trabalhos envolvendo a toxicidade por prótons são escassos. A sensibilidade de células a pH baixo depende da fase de crescimento e desenvolvimento em que estas se encontram. Em raízes, as células mais sensíveis são as da região de alongamento e em suspensões celulares as células na fase log de crescimento são mais sensíveis do que as células na fase estacionária. Este trabalho faz parte de uma linha de pesquisa que procura explorar as diferenças que existem entre células quanto à sensibilidade a pH baixo para melhor entender a toxicidade e tolerância a prótons. Foram utilizadas células da cultura de tabaco cv. BY-2, um sistema modelo que apresenta diversas vantagens sobre o uso de raízes para a realização deste trabalho. Os objetivos deste estudo foram de a) determinar condições apropriadas para a exposição destas células a acidez; b) caracterizar a resposta destas células a pH baixo, sob influência de diferentes fatores ambientais, quando se encontram em estados distintos de sensibilidade ao pH baixo; e c) verificar se mudanças na sensibilidade das células a pH baixo podem ser decorrentes de alterações na composição da membrana plasmática ou na atividade das ATPases. Vários testes iniciais foram realizados com o intuito de se definir algumas condições experimentais - o tempo de exposição, a composição do meio e o tampão a ser empregado. Optou-se por lavar as células e depois incubá-las por 1h em meio simples com pH desejado e contendo apenas Ca \'Cl IND. 2\' 2mM, KCl 10mM. E o tampão MES ou biftalato (10 mM). O biftalato foi testado porque o tampão MES, usado normalmente no meio de cultura completo, não é eficiente na faixa de pH abaixo de 5,0. O biftalato (pKa = 4,1) praticamente não afetou a viabilidade celular avaliada pela permeabilidade a trypan blue, mas inibiu o crescimento celular no meio de cultura completo. Mesmo assim, os dois tampões foram utilizados em paralelo em diversos experimentos e verificou-se que os resultados foram semelhantes. A viabilidade das células na fase log (2 dias) foi reduzida quando se abaixou o pH de 5,6 a 3,8, sendo que caiu mais acentuadamente até o pH de 4,8. As células da fase estacionaria (7d) foram insensíveis ao baixo pH. A um pH fixo de 4,2 aumentando-se a concentração de Ca \'Cl IND. 2\' para cerca de 8 a 16 mM praticamente aboliu o efeito deletério do pH baixo. Para se ter o mesmo efeito com a adição de KCl, foi necessário adicionar entre 80 e 160 mM. A adição de sacarose também amenizou os efeitos do pH\'sendo praticamente revertido a uma concentração de 100 Mm. Os resultados indicam a importância da força iônica e da osmolaridade da solução, mas o efeito de Ca não parece depender apenas destas duas propriedades. A inibição de ATPases, pelo uso de DCCD, não pareceu ter qualquer relação com a sensibilidade a pH baixo. Tanto células de tabaco na fase log quanto estacionárias foram sensíveis à aplicação de ortovanadato de sódio. Em células da fase estacionária, este efeito mais acentuado a pH 4,2, sugerindo que nestas células, as H+ ATPases do tipo P da membrana plasmática podem ter algum papel na tolerância destas células ao baixo pH. Encontrou-se diferenças no perfil protéico de frações enriquecidas em membrana plasmática entre células da fase log e estacionárias e entre células tratadas ou não a pH baixo. Estas diferenças precisam ser melhor estudadas. / Acid soils account for about 40 % of the surface area of the world and are one of the major factors limiting plant productivity. In Brazil, these soils comprise roughly two-thirds of its total territory. In general, soil acidity is detrimental because it limits the depth of the root system, increasing susceptibility to drought and decreasing the use of nutrients. In addition to the high levels of hydrogen ion activity, acid soils usually have Al toxicity hazards, a topic which has been intensely studied in the past years. However, Al toxicity only occurs at low pH, under conditions in which proton toxicity is also a problem. Despite this, studies of proton toxicity are lacking. The sensitivity of cells to low pH depends on their growth and developmental status. In roots, the most sensitive cells are those of the elongation zone and in cell cultures, cells in the log phase are more sensitive than those of the stationary phase. This study is part of a larger attempt to explore the differences that exist between cells with respect to their sensitivity to low pH to further understand toxicity and tolerance to protons. Cells of tobacco BY-2, a plant cell model system which has several advantages over the use of roots, was used in this study. The objectives were a) to determine the appropriate conditions to expose these cells to low pH; b) characterize the response of these cells to low pH, when different environmental factors are varied and at different stages of cellular sensitivity to acidity; and c) examine if changes in the sensitivity of cells to low pH are due to changes in the composition of plasma membranes or in the activity of ATPases. Several preliminary tests were performed to define the experimental conditions to be used ? the duration of exposure to low pH, the composition of the medium and the buffer to be used. A simple solution containing only CaCl2 2mM, KCl 10 mM and MES or phthalate buffer (10 mM) was chosen to wash and incubate the cells at the pH of interest. Phthalate was tested because MES, the buffer normally used in the culture medium, is not effective at pH values below 5.0. Phthalate (pKa = 4,1) had very little effects on cell viability as evaluated by membrane permeability to trypan blue, but it severely inhibited the growth of the cell culture in complete medium. Nevertheless, both buffers were used in several subsequent experiments, the results of which were found to be similar between both buffers. The viability of log-phase cells (2 day-old) was reduced when the pH was lowered from pH 5,6 to 3.8, but this was sharper down to pH 4,8. Cells in stationary phase (7 day-old) were insensitive to low pH. At a fixed pH of 4,2, increases in the concentration of CaCl2 up to 8 or 16 mM abolished most of the deleterious effects of low pH. To generate the same effect, KCl had to be added at concentrations between 80 and 160 mM. The addition of sucrose also alleviated the effects of low pH. The results suggest the importance of solution ionic strength and osmolarity on sensitivity to low pH, but the effects of Ca do not appear to depend only on these two properties. The inhibition of ATPases, by DCCD, did not appear to bear any relation to cellular sensitivity to low pH. Both log-phase and stationary-phase cells were affected by the addition of sodium orthophosphate. In stationary-phase cells, this effect was more pronounced at pH 4,2, suggesting that in these cells, P-type H+-ATPases of the plasma membrane may play a role in the tolerance of these cells to lo pH. Differences were found in the protein profile of enriched plasma membrane fractions both between log-phase and stationary-phase cells and between cells treated or not with low pH. These differences, however, need to be better examined.

Master of Science / Department of Animal Sciences and Industry / Teresa L. Douthit / Abruptly increasing concentrate in the ration of horses results in altered cecal dynamics which can culminate in digestive distress. Nine Quarter horses previously fitted with cecal cannulae were utilized for 3 consecutive 22-d experiments, each separated by 2 d of rest. During Exp. 1 and 2 horses were acclimated to the same ration for the initial 21 d of each period, followed by a concentrate challenge on d 22. The acclimation ration consisted of a morning meal of 0.5% BW concentrate (Omolene 200, Purina Animal Nutrition, LLC, Gray Summit, MO) fed with 1.5% BW prairie grass hay divided evenly between a morning and evening meal. On d 22 of Exp. 1, horses were fed a morning meal consisting solely of 1.0% BW concentrate while 1.25% BW concentrate was fed on d 22 of Exp. 2. Cecal samples were obtained through cecal cannulae from d 19 to 22 of each experiment every 4 h for h 24 following the morning meal each day. Cecal pH during Exp. 1 was recorded and decreased at h 12 following the concentrate meal on d 22 in comparison to cecal pH at h 12 on d 19 to 21 (P = 0.009). During Exp. 2 cecal pH increased at h 4 (P = 0.02) and decreased at h 12 and 20 (P < 0.0001) following this oncentrate challenge compared to cecal pH recorded at the same time points during the acclimation period. Experiment 3 differed from that of Exp. 2 only in the respect that during the acclimation period horses were fed, in addition to the acclimation ration, either a proprietary supplement (n = 5) or a placebo (n = 5). Cecal samples from d 19 to 22 were analyzed for pH, concentration of lactate, and concentration of VFA. Horses consuming the supplement had increased cecal pH at h 4 (P = 0.009), concurrently decreased cecal lactate (P = 0.02), increased ratio of (acetate+butyrate)/propionate at h 8 and 16 (P ≤ 0.006), and decreased VFA concentration at h 24 (P ≤ 0.05) compared to horses in the control group following the concentrate challenge.

Sustainable rehabilitation can be compromised by the inability of vegetation to survive in hostile mine wastes on a long-term basis. The adverse chemical and physical properties of mine wastes, together with extreme pH conditions and lack of nutrients, provide poor growth conditions for vegetation during seed development and germination. This raises concern for the long-term survival of vegetation through means of seed production when under stress from the punitive properties of mine wastes. Seed vigour is a function of a variety of factors to which the parent plant is subjected during seed formation and maturation. Environmental conditions experienced by the maternal plant during the growth season plays a significant role in determining subsequent germination rates in seeds. Traits of offspring seed depend on the abiotic environment attributed by the growth medium during seed development and maturation The general aim of this study was to determine the viability of seed produced by a previous generation of grass species established in eight different mine wastes and two soils (namely: gypsum wastes; gold tailings with low pyrite content; gold tailings with high pyrite content; platinum tailings; kimberlite mine waste; fluorspar mine waste; andalusite mine waste; coal discard; red soil; and vertic soil) in order to identify suitable species for specific mine wastes to ensure long-term survival through means of seed production. The species selected included: Eragrostis curvula; Eragrostis tef; Cenchrus ciliaris; Eragrostis curvula; Digitaria eriantha; Cynodon dactylon; Chloris gayana; Hyparrhenia hirta; and Sorghum bicolor. The progeny seed‟s viability and ability to germinate were determined through a pot trial study and additional germination testing at the laboratory of Advance Seed (Pty) Ltd. (AS). The germination results were correlated with the growth media analyses by statistical non-parametric correlations which indicated several significant correlations among the growth media properties themselves, and with the germination of the progeny seed. C. gayana (Rhodes grass) seed had poor germination percentages, especially seed harvested from Rhodes grass grown in acidic wastes. Seed harvested from each of the E. curvula grasses grown in various mine wastes, had excellent germination percentages. According to the Repeated Measures ANOVA statistical analysis, there was a significant influence of the growth media in which the parent grass were grown as a variable on the germination of the progeny seed batches from S. bicolor, C. ciliaris, C. gayana, and D. eriantha, indicating that the environmental factors as attributed by the growth media, i.e. the eight different mine waste materials and two soils, and experienced by the maternal plant, did indeed influence the germination of progeny seed. However, it was found that significant correlations between the properties of the growth media and the germination of the progeny seed, was species dependent. The second general aim for this study was to evaluate above-ground re-growth of parent plants after cutting in the mine waste materials and soil types mentioned above. The ability of established grasses to re-grow after a cutting event was determined by cutting the above-ground biomass of the parent grasses, after which it was scored according observable above-ground growth in the following growth season. The measurement of re-growth was subjectively done by scoring the grasses according to observable above-ground biomass. Re-growth was observed for all the perennial grass species. This can be ascribed to the grasses showing resilience to stress factors attributed by the growth media; or new grasses which emerged from seed that collected in the pots, being mistaken for re-growth; or new emerging grasses from the nodes of stolons and/or rhizomes being mistaken for re-growth. However, the emergence of new grasses was an indicator of good health, as biomass allocation to rhizomes and stolons is reduced under low nutrient availability and stress conditions. Therefore the emergence of new grasses is indicative that the plant is either tolerant to stress conditions or that the plant adapted to the restriction of growth due to the roots being bound to the size of the pot. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015