A biofilter removes biodegradable contaminants from air by passing it through a biologically-active packed bed. The microorganism community is of fundamental interest but has been the focus of few studies. This work is an investigation of the bacterial community based on the potential functional diversity of the community. <br /><br /> A number of experiments were performed in laboratory-scale biofilters using ethanol as a model contaminant. All biofilters were able to remove the ethanol with elimination capacities in the range 80 to 200gVOCm^-3h^-1; these values are comparable with published literature. Natural organic media (peat or compost) was used as packing. <br /><br /> The potential functional diversity of the community was assessed by Community-Level Physiological Profiling (CLPP) using sole-Carbon Source Utilisation Profile (CSUP). Community samples were used to inoculate Biolog EcoPlates^TM: microplates containing a selection of 31 different carbon-substrates and an indicator dye responding to bacterial growth. This technique was found to be sensitive to changes in the community structure over time and location. <br /><br /> Results showed that the community in samples taken close together (over a scale of a few centimetres) are similar and that relatively small media samples (0. 5 to 1 g) provide reproducible information. A study of a single biofilter indicated stratification of the community occurring with the community near the inlet diverging from that near the middle and outlet of the unit; this is attributed to the ethanol being degraded in the upper part of the column and the lower part of the column not being subjected to ethanol loading. In a study of two units at a higher loading rate, stratification was not observed over a period of weeks; it is suggested that the stratification may develop over this timescale as a result of the presence or absence of the Volatile Organic Compound (VOC) and not due to differences in concentration. <br /><br /> An acclimation period of 7 to 10 days was observed before near-complete removal of ethanol was attained. Monitoring of the community suggested a subsequent shift in diversity. It is suggested that the initial acclimation period is due to biofilm formation and the subsequent shift in community diversity is due to re-organisation of the community as species specialise. In a portion of the biofilter with minimal ethanol exposure, a sudden shift in community is observed after a period of some weeks. This may reflect changes as a result of starvation and indicates that periods of shut-down (when the biofilter is not loaded) may affect the community. <br /><br /> Two studies of biofilters operating in parallel were carried out. The first provided evidence of a divergence in the communities over a period of two weeks. In the second, communities in the two units underwent changes over time but observations from both units at any one time were similar. This demonstrates that biofilters set-up and operated in a similar manner may maintain similar communities but that this is not necessarily the case. This has implications for the reproducibility of laboratory experiments and for the variation of community structure with horizontal position in industrial units.

A biofilter removes biodegradable contaminants from air by passing it through a biologically-active packed bed. The microorganism community is of fundamental interest but has been the focus of few studies. This work is an investigation of the bacterial community based on the potential functional diversity of the community. <br /><br /> A number of experiments were performed in laboratory-scale biofilters using ethanol as a model contaminant. All biofilters were able to remove the ethanol with elimination capacities in the range 80 to 200gVOCm^-3h^-1; these values are comparable with published literature. Natural organic media (peat or compost) was used as packing. <br /><br /> The potential functional diversity of the community was assessed by Community-Level Physiological Profiling (CLPP) using sole-Carbon Source Utilisation Profile (CSUP). Community samples were used to inoculate Biolog EcoPlates^TM: microplates containing a selection of 31 different carbon-substrates and an indicator dye responding to bacterial growth. This technique was found to be sensitive to changes in the community structure over time and location. <br /><br /> Results showed that the community in samples taken close together (over a scale of a few centimetres) are similar and that relatively small media samples (0. 5 to 1 g) provide reproducible information. A study of a single biofilter indicated stratification of the community occurring with the community near the inlet diverging from that near the middle and outlet of the unit; this is attributed to the ethanol being degraded in the upper part of the column and the lower part of the column not being subjected to ethanol loading. In a study of two units at a higher loading rate, stratification was not observed over a period of weeks; it is suggested that the stratification may develop over this timescale as a result of the presence or absence of the Volatile Organic Compound (VOC) and not due to differences in concentration. <br /><br /> An acclimation period of 7 to 10 days was observed before near-complete removal of ethanol was attained. Monitoring of the community suggested a subsequent shift in diversity. It is suggested that the initial acclimation period is due to biofilm formation and the subsequent shift in community diversity is due to re-organisation of the community as species specialise. In a portion of the biofilter with minimal ethanol exposure, a sudden shift in community is observed after a period of some weeks. This may reflect changes as a result of starvation and indicates that periods of shut-down (when the biofilter is not loaded) may affect the community. <br /><br /> Two studies of biofilters operating in parallel were carried out. The first provided evidence of a divergence in the communities over a period of two weeks. In the second, communities in the two units underwent changes over time but observations from both units at any one time were similar. This demonstrates that biofilters set-up and operated in a similar manner may maintain similar communities but that this is not necessarily the case. This has implications for the reproducibility of laboratory experiments and for the variation of community structure with horizontal position in industrial units.

Thesis (M.Sc.)--Acadia University, 2001. / Includes bibliographical references (leaves 62-64). Also available on the Internet via the World Wide Web.

Visible lesions on coral colonies are potential indicators that environmental stressors are influencing a reef. To test this hypothesis, pairs of near-shore reefs on Taiwan were surveyed along an anthropogenically influenced gradient that included locations near the cities of Taipei and Taitung, and more remote reefs off Green Island. Two fringing reefs at Sanya, Hainan Island, a popular Chinese resort area, were also assessed. Field surveys were undertaken to detect, quantify and visually describe the occurrence of lesions at each site. Coral mucus samples were collected from both normal-appearing polyps and lesion-afflicted areas of colonies to assess carbon requirements of associated microbes. Tissue samples were also collected to identify bacterial communities inhabiting healthy tissue for comparison with those associated with lesions; denaturing gradient gel electrophoresis and 16S rRNA sequencing for bacterial identification were utilized in these analyses. In addition, tissue samples were collected in the vicinity of lesions and prepared for histological examination. At sites in Taiwan, lesions were encountered twice as often at the sites near Taipei and Taitung than at Green Island. The fewest (15/72 sightings) lesions were encountered at the reefs near Sanya, primarily because there has been nearly an 80% loss of coral cover at Sanya in recent decades. Overall, tissue loss was the most common lesion recorded (52%), followed by pink discoloration (27%) and color loss (i.e., bleaching, 15%). Porites was the taxon most commonly observed with one or more lesions (45% of sightings). Microbes within mucus from lesioned areas utilized similar carbon sources as microbes from mucus from healthy polyps, but utilized those sources more than twice as often. Examples of carbon sources utilized by microbes in >50% of the lesion samples were D-cellobiose, D-mannitol, N-acetyl-D-glucosamine, alpha-cyclodextrin, and glycogen. Bacterial assemblages on corals were significantly different between Taiwan and China, among sites, and between water samples and coral samples, but not between healthy samples and lesions. Bacterial sequences identified in tissue samples from lesions revealed the presence of well-known disease-related genera, such as Clostridium and Vibrio. Microbes specifically indicating anthropogenic sources, included Bacillus sp. (sewage sludge) and Geobacillus thermolevorans (irritable bowel syndrome). Histological examination of tissue samples, particularly those from lesions characterized as tissue loss, revealed fragmentation and detachment from the mesoglea of gastrodermis and epidermis, as well as brown granular material, and the presence of ciliates and small crustaceans. Corals are susceptible to a variety of diseases. For reefs in the western Atlantic and Caribbean, occurrences of lesions and characterization of coral diseases have been relatively well documented. In contrast, many areas in the vast Indo-Pacific, including the reefs of Taiwan and China, have received much less attention. This study of lesions and associated microbiomes on nearshore reefs of Taiwan and Hainan Island supports previous research that has revealed higher incidences of coral lesions and disease in reefs near extensive human populations. The results also support the hypothesis that many of the microbes associated with coral lesions are part of the natural coral microbiome and that some microbes can become opportunistic when the host corals are stressed.

Sound research and effective teaching are both essential to the progress of science. This thesis encompasses two studies to address the two needs: a multi-scale soil study designed to validate a novel soil biological characterization method; and a pilot pedagogical study designed to test the efficacy of authentic science inquiry into gas chromatography. The soil study relies on a comparison of six soils taken from the Bahamas and Michigan. The novel method, using soil-derived VOCs analyzed via gas chromatography-mass spectrometry (GC-MS), proved effective for resolving soils, as hypothesized, and may prove useful for analyzing soil biology rapidly and non-destructively in future studies. The pilot pedagogical study compares traditional recipe-style instruction with authentic science inquiry in an undergraduate chemistry laboratory class. Pre- and post-assessments of students’ conceptual understanding, retention of terms, and attitude revealed the hypothesized superior efficacy of authentic science inquiry over traditional recipe-style instruction.

No description available.

Maize is the staple food for a majority of people in Southern Africa, but plant diseases are responsible for at least 10% of crop production losses. Bacterial leaf streak (BLS) of maize was first reported in South Africa in 1949 and has not been reported elsewhere. Very little is known about the pathogen involved and therefore it is deemed necessary to compile a characteristic profile for the pathogen to prevent the possibility of major crop losses as a result of this disease. This study aimed to use biochemical and phenotypic methods to determine the specific characteristics of the causal agent of BLS. Diseased plant material showing symptoms of BLS were collected during the maize production seasons of 2012 and 2013 within South Africa’s maize production regions namely the North West, Free State, Gauteng and Northern Cape provinces. To prevent contamination, maize leaves were surface sterilised thoroughly before bacterial isolation commenced. Sections of the infected maize leaves were placed on GYC agar plates on which yellow, mucoid bacterial colonies after incubation for 24 to 48 hrs. The isolated bacteria were purified and the molecular identification of the bacteria was conducted in a related study. Although literature indicates that Xanthomonas campestris pv. zeae is the causal agent of BLS, pure cultures obtained from maize leaves showing characteristic symptoms of BLS were identified as species of Xanthomonas, Pantoea, and Enterobacter. To elucidate the pathogenicity of the isolated strains, pathogenicity tests based on Koch’s postulates were performed. Results from the pathogenicity tests confirmed that only the isolate Xanthomonas species was capable of inducing the characteristic BLS symptoms when healthy maize plants were inoculated with the suspected pathogens. It is important to inoculate the maize seedlings at the correct age (four-leaf stage) and the spray method is recommended. Re-isolation was repeated from the same plant material used during the initial isolation process but the isolation method was amended. The optimised isolation method involved the use of a dilution range and spread plate method. Colonies from this isolation technique grew as bright yellow colonies that were identified as Xanthomonas spp. This outcome indicates the importance of surface sterilisation, pulverisation and subsequent dilution of plant materials for isolation of bacterial pathogens from diseases plants. These isolates were used to create protein profiles with SDS-PAGE electrophoresis and carbon utilisation patterns with the Biolog® GN2 system. Protein profiling banding patterns was assessed based on presence/absence criteria. Highly similar protein profiles were observed among the X. campestris pv. zeae isolates but groupings of different protein profiles were determined when minor differences in the protein profiles was taken into account. Xanthomonas campestris pv. zeae was successfully distinguished from the X. axonopodis pv. vasculorum reference strain through unique SDS banding patterns. Banding patterns obtained from cultures grown in a liquid medium (tryptic soy broth) were of a higher quality than the banding patterns obtained from bacteria harvested from solid media (CYG agar plates). Carbon source utilisation data was used to evaluate the average well colour development obtained from each isolate. Statistically significant differences were found among some of the isolates, with some isolates being metabolically more active than other isolates. Substrate utilisation patterns produced by the isolates corresponded to previously published studies on various Xanthomonas species. The cell count of the samples used during carbon utilisation patterns must be standardised in order to obtain reliable results. During this study, the application of Koch’s postulates and two inoculation techniques confirmed that Xanthomonas campestris pv. zeae is the pathogen responsible for bacterial leaf streak of maize. Members of the Pantoea and Enterobacter genera were found on the leaf surface of maize plants infected with BLS but inoculations of healthy maize plants with these bacteria did not result in bacterial leaf streak symptoms on the maize plants. These bacteria were not pathogenic and were considered endophytes. The identified pathogen was characterised through protein and metabolic profiling. The protein profiles of the pathogen obtained through analysis of the major bands of the SDS-PAGE gels were highly similar and distinguishable from the Xanthomonas reference culture. Groupings within the X. campestris pv. zeae group was found when major and minor bands were considered, this may however be altered when the intensities of the bands are used during analysis. Carbon utilisation patterns were assessed using Biolog® GN2 plates. A metabolic fingerprint was created for the pathogen of BLS, it was possible to distinguish between X. campestris pv. zeae and other Xanthomonas strains based on the fingerprint. This fingerprint could be used to identify the pathogen. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Maize is the staple food for a majority of people in Southern Africa, but plant diseases are responsible for at least 10% of crop production losses. Bacterial leaf streak (BLS) of maize was first reported in South Africa in 1949 and has not been reported elsewhere. Very little is known about the pathogen involved and therefore it is deemed necessary to compile a characteristic profile for the pathogen to prevent the possibility of major crop losses as a result of this disease. This study aimed to use biochemical and phenotypic methods to determine the specific characteristics of the causal agent of BLS. Diseased plant material showing symptoms of BLS were collected during the maize production seasons of 2012 and 2013 within South Africa’s maize production regions namely the North West, Free State, Gauteng and Northern Cape provinces. To prevent contamination, maize leaves were surface sterilised thoroughly before bacterial isolation commenced. Sections of the infected maize leaves were placed on GYC agar plates on which yellow, mucoid bacterial colonies after incubation for 24 to 48 hrs. The isolated bacteria were purified and the molecular identification of the bacteria was conducted in a related study. Although literature indicates that Xanthomonas campestris pv. zeae is the causal agent of BLS, pure cultures obtained from maize leaves showing characteristic symptoms of BLS were identified as species of Xanthomonas, Pantoea, and Enterobacter. To elucidate the pathogenicity of the isolated strains, pathogenicity tests based on Koch’s postulates were performed. Results from the pathogenicity tests confirmed that only the isolate Xanthomonas species was capable of inducing the characteristic BLS symptoms when healthy maize plants were inoculated with the suspected pathogens. It is important to inoculate the maize seedlings at the correct age (four-leaf stage) and the spray method is recommended. Re-isolation was repeated from the same plant material used during the initial isolation process but the isolation method was amended. The optimised isolation method involved the use of a dilution range and spread plate method. Colonies from this isolation technique grew as bright yellow colonies that were identified as Xanthomonas spp. This outcome indicates the importance of surface sterilisation, pulverisation and subsequent dilution of plant materials for isolation of bacterial pathogens from diseases plants. These isolates were used to create protein profiles with SDS-PAGE electrophoresis and carbon utilisation patterns with the Biolog® GN2 system. Protein profiling banding patterns was assessed based on presence/absence criteria. Highly similar protein profiles were observed among the X. campestris pv. zeae isolates but groupings of different protein profiles were determined when minor differences in the protein profiles was taken into account. Xanthomonas campestris pv. zeae was successfully distinguished from the X. axonopodis pv. vasculorum reference strain through unique SDS banding patterns. Banding patterns obtained from cultures grown in a liquid medium (tryptic soy broth) were of a higher quality than the banding patterns obtained from bacteria harvested from solid media (CYG agar plates). Carbon source utilisation data was used to evaluate the average well colour development obtained from each isolate. Statistically significant differences were found among some of the isolates, with some isolates being metabolically more active than other isolates. Substrate utilisation patterns produced by the isolates corresponded to previously published studies on various Xanthomonas species. The cell count of the samples used during carbon utilisation patterns must be standardised in order to obtain reliable results. During this study, the application of Koch’s postulates and two inoculation techniques confirmed that Xanthomonas campestris pv. zeae is the pathogen responsible for bacterial leaf streak of maize. Members of the Pantoea and Enterobacter genera were found on the leaf surface of maize plants infected with BLS but inoculations of healthy maize plants with these bacteria did not result in bacterial leaf streak symptoms on the maize plants. These bacteria were not pathogenic and were considered endophytes. The identified pathogen was characterised through protein and metabolic profiling. The protein profiles of the pathogen obtained through analysis of the major bands of the SDS-PAGE gels were highly similar and distinguishable from the Xanthomonas reference culture. Groupings within the X. campestris pv. zeae group was found when major and minor bands were considered, this may however be altered when the intensities of the bands are used during analysis. Carbon utilisation patterns were assessed using Biolog® GN2 plates. A metabolic fingerprint was created for the pathogen of BLS, it was possible to distinguish between X. campestris pv. zeae and other Xanthomonas strains based on the fingerprint. This fingerprint could be used to identify the pathogen. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

No description available.

Os microrganismos representam a forma de vida mais abundante e diversificada do planeta. A atividade agrícola leva a uma redução da biodiversidade do solo e a menor diversidade microbiana pode resultar na diminuição da ciclagem de nutrientes e no crescimento das plantas. Como forma de avaliar alterações na atividade microbiana e na estrutura das comunidades de bactérias do solo, decorrentes do cultivo da cana-de-açúcar, foram conduzidos dois experimentos. O primeiro, no município de Novo Horizonte (SP), com o objetivo de determinar ação da queima da cana-de-açúcar sobre a comunidade de bactérias do solo e o segundo experimento, nos municípios de Pirassununga (SP) e Jaboticabal (SP), com o objetivo de verificar o efeito da adubação nitrogenada sobre a comunidade bacteriana do solo. Amostras de terra foram coletadas nas profundidades 0-10 e 10-20 cm, na linha e entrelinha de plantio. No primeiro experimento foram utilizadas três cultivares de cana-de-açúcar (SP81-3250, SP80-1842 e RB72-454) sob os sistemas de manejo de colheita sem queima (mecanizada) e com queima (manual) prévia a colheita. Nesse experimento foram avaliadas a diversidade metabólica (Biolog) e a estrutura das comunidades bacterianas por meio da PCR-DGGE do gene rRNA 16S. A mudança no manejo de colheita da cana-de-açúcar provocou modificações no metabolismo heterotrófico do solo, alterando a diversidade metabólica. No entanto, não houve mudanças na estrutura das comunidades bacterianas do solo com e sem queima sob a variedade SP801842. Dessa forma, a primeira queima da cana-de-açúcar previamente à colheita alterou a capacidade e a diversidade metabólica microbiana, mas não mudou a estrutura das comunidades de bactérias em relação à área sem queima. No segundo experimento foram avaliadas amostras de terra, de duas áreas experimentais, sob cultivo de cana-de-açúcar (SP813250), com diferentes doses de N (0, 40, 80 e 120 kg de N ha-1) na forma de uréia, aplicadas no sulco de plantio. Para verificar possíveis alterações na comunidade bacteriana desses solos, foram avaliadas a estrutura das comunidades bacterianas por PCR-DGGE e a diversidade de bactérias oxidadoras de amônio (AOB), pelo seqüenciamento de bibliotecas do gene rRNA 16S, utilizando oligonucleotídeos iniciadores específicos. As doses de N alteraram a estrutura das comunidades bacterianas do solo nas duas áreas experimentais, determinadas por PCR-DGGE, entretanto, a adubação nitrogenada não alterou a diversidade de AOB no solo, das duas áreas. A estrutura da comunidade de AOB no solo da USA, sem adubação nitrogenada e com 80 kg de N ha-1 diferiu estatisticamente. Nas duas áreas, as unidades taxonômicas operacionais mais abundantes se relacionam filogeneticamente a Nitrosospira multiformes. / The microorganisms are the most abundant and diverse living creatures on earth. The agricultural practices reduce the soil biodiversity and a lower microbial diversity can result in a nutrient cycling and plant growth reduction. Two sugarcane crops experiments were investigated to evaluate modifications in the microbial activity and soil bacterial communities structure. The first of them was done at municipality of Novo Horizonte, Sao Paulo State (SP), and it has the aim to determine the sugarcane burn effects on soil bacterial community. The second experiment was introduced at municipalities of Pirassununga (SP) and Jaboticabal (SP) with the objective to verify the nitrogen fertilizing effect on soil bacterial community. The soil samples were taken in 0-10 cm and 10-20 cm depth, between and in the planting furrows. We used three sugarcane varieties (SP81-3250, SP80-1842 e RB72-454) in the first experiment under unburned and burned sugarcane pre-harvest. We evaluated metabolic diversity (Biolog) and the bacterial community structure performing PCR-DGGE of 16S rRNA gene in the first experiment. The sugarcane harvest management had modified the soil heterotrophic metabolism by altering its diversity. In spite of that, there is no difference between the burned and unburned soil bacterial communities under the variety SP80-1842. For that reason, the first year pre harvest burn altered the microbial metabolic diversity and capacity but did not change the bacterial community structure when related with unburned area. In the second experiment, soil samples under the SP81-3250 variety were analyzed from two sites. Each site received different levels of urea as nitrogen fertilization (0, 40, 80 e 120 kg de N ha-1) applied at planting furrows. The PCR-DGGE was applied to verify changes in the bacterial communities structure in these soils. The ammonia-oxidizing bacteria (AOB) diversity was evaluated by sequencing of 16S rRNA gene libraries after the amplification with specific primers. The levels of nitrogen fertilization altered the soil bacterial communities structure in both study sites, by PCRDGGE evaluation. However, the nitrogen application did not alter the soil AOB diversity in those two sites. The soil AOB community structure under no nitrogen and 80 kg N ha-1 application was different from the community structure under the other levels of fertilization in one of the two sites. The operational taxonomic unit are phylogenetically related to Nitrosospira multiformes in the two sites.