Salmonella enterica infection affects a wide range of animals including humans. The avian specific serotype S. Pullorum infection produces systemic disease followed by a persistent carrier state in convalescence birds. Vaccination and other control strategies require an improved understanding of the immunity in response to S. Pullorum infection. This study compared the different immune dynamics following infection with (persistent) S. Pullorum and related (non-persistent) serovars S. Enteritidis and S. Gallinarum using co-culture of Salmonella-infected macrophages and CD4+ T lymphocytes in vitro and 2-day-old chickens in vivo. In comparison with S. Enteritidis, macrophages infected with S. Pullorum had a reduced gene expression of pro-inflammatory cytokines CXCLi2, IL-6, iNOS, IFN-γ, IL-12α and IL-18 and lower level of nitrite production. S. Pullorum-infected macrophages were found to be less effective than S. Enteritidis in stimulating the CD4+ lymphocytes to proliferate in vitro. CD4+ lymphocytes in co-culture with Salmonella-infected macrophages also produced lower levels of IFN-γ and IL-17F mRNA in response to S. Pullorum compared with S. Enteritidis. S. Pullorum infection in 2-day-old chickens stimulated proliferation of Th2-like lymphocytes with reduced IFN-γ and IL-17F but increased IL-4, IL-13 and IL-10 in the caecal tonsils and spleens when compared to S. Enteritidis. However, the modulation by S. Pullorum is not likely to be related to its large virulence plasmid, although the virulence plasmid of S. Gallinarum was shown to reduce nitrite production and gene expression of IL-1β and iNOS in infected HD11 cells. Our data showed no evidence of clonal anergy or immune suppression induced by S. Pullorum in vitro. The experimental work thus shows that the response to S. Pullorum infection was characterised by a modulation on host immunity from a dominant IFN-γ-producing Th17 response towards a Th2-like response which may promote persistent infection in chickens. This study provides insights into mechanisms by which S. Pullorum evades host immunity and produces the persistent carrier state. This opens the possibility for therapeutic application of cytokines to restore the host protective immune response to eliminate infection.

A crescente eutrofização dos ambientes aquáticos é consequência das atividades humanas que desequilibram os ecossistemas. A descarga de esgotos urbanos, a utilização de adubos químicos, os efluentes das agroindústrias e dos diversos setores industriais promovem a entrada de quantidades significativas de nutrientes e matéria orgânica em corpos d´água, favorecendo o aumento das florações de algas, entre elas as cianobactérias. O excesso de matéria orgânica e os despejos de efluentes agroindustriais nos mananciais hídricos representam os principais fatores do excesso de floração das cianobactérias, destacando-se a Microcystis aeruginosa e da sua cianotoxina microcistina. Nesta tese foi aplicado o Processo Oxidativo Avançado (POA) de Fotoeletroxidação (FEO) para a degradação de Microcystis aeruginosa e da cianotoxina microcistina. Investigou-se a otimização de parâmetros como tempo de tratamento e intensidade de corrente elétrica aplicada. Os resultados obtidos nas condições aplicadas demonstram valores de 99% para a degradação de Microcystis aeruginosa e cianotoxina Microcistina com o tempo de 10min e densidade de corrente de 2mA.cm-2. Os testes de toxicidade aguda com ajuste de pH de 6,3 para 7,3 não apresentaram toxidade para a espécie teste Pimephales promelas. / The increasing eutrophication of aquatic environments is a consequence of human activities that disrupt ecosystems. The discharge of urban sewage, the use of chemical fertilizers, effluents from agro-industries and other industrial sectors promote the input of significant amounts of nutrients and organic matter in water, favoring increased blooms of algae, including cyanobacteria. Excessive organic matter and the discharge of agroindustrial effluents in the water bodies, represent the main factors for the excessive bloom of cyanobacteria, especially Microcystis aeruginosa and microcystin cianotoxin. In this study, an Advanced Oxidation Process named photoelectrooxidation, was used to the degradation of Microcystis aeruginosa and cianotoxin microcystin. The optimization of parameters such as, treatment time and intensity of electrical current applied in the process of PEO were evaluated. The results achieved under the conditions applied, show 99% of degradation with experimental times at 10 minutes and a current density of 2mA.cm-2. The acute toxicity tests, with pH adjustment from 6.3 to 7.3, did not show toxicity to the species Pimephales promelas.

Cyanobacteria, also known as blue-green algae, are known to produce a number of biologically active compounds. Extracts of cultured cyanobacteria isolated from South Florida sources were screened for possible developmental toxins using the zebrafish (Danio rerio) embryo as a model of vertebrate development. A strain of cyanobacteria, Fischerella 52-1, isolated from the Florida Everglades, was found to produce metabolites that caused a consistent developmental dysfunction in embryos exposed to lipophilic extract. Initial chemical characterization of the bioactive fraction identified a series of eight apparent indole-containing compounds. The two main components were purified using the zebrafish embryo model to guide the fractionation. Chemical characterization using 1- and 2-dimensional NMR, HESIMS, HRHESIMS, and IR determined that the two main compounds were the previously identified 12-epi-Hapalindole H Isonitrile, and a novel compound 12-epi-Ambiugine B Nitrile. The major contributor of the developmental defects detected in the zebrafish embryos was 12-epi-Hapalindole H Isonitrile.

<p>Exposure to ultraviolet radiation (UVR) has been shown to cause cellular damage in cyanobacteria. In response to UVR exposure, some cyanobacteria produce scytonemin, an indole-alkaloid sunscreen capable of absorbing long-wavelength UVA radiation. Previous genomic and transcriptomic analyses have determined that the production of scytonemin is controlled by a two-component regulatory system (TCRS), encoded by Npun_F1277 and Npun_F1278 in the cyanobacterium <i>Nostoc punctiforme </i>ATCC 29133. This TCRS is thought to not only regulate scytonemin biosynthesis, but also other responses to light and UVR stimuli. To better understand the functionality of the sensor kinase (SK) Npun_F1277 and to determine if it could activate alternative UVR protection pathways, the SK was expressed in <i>Escherichia coli.</i> The first objective of this study was to observe and quantify the level of fitness conferred to <i>E. coli</i> expressing Npun_F1277 from <i>N. punctiforme </i>(strain SKE) when exposed to white light, UVA, and UVB stress. Results from these experiments do not indicate that expression of the <i>N. punctiforme</i> SK conferred an advantage to <i>E. coli</i> under white light, UVA, or UVB stress based on growth alone. Therefore, the second objective was to study the expression of regulatory genes, such as response regulators, in <i>E. coli</i> that are homologs to those associated with the SK Npun_F1277 in <i>N. punctiforme </i>using quantitative-PCR. Expression of the selected genes was measured following exposure to white light and UVA after 30 and 60 minutes as well as UVB after 15 and 30 minutes. Comparison of SKE to empty-vector (EV) control cells exposed to the same stress showed that there were significant changes in the expression of important regulatory genes (e.g. <i>recA, spoT, relA</i>) in the SKE strain. Moreover, when comparing SKE cells exposed to the same conditions above to unstressed SKE cells, a similar result was seen for SKE cells exposed to UVA and UVB as was found in the studies comparing SKE to EV cells. These results suggest that the SK Npun_F1277 may play a role in multiple defense mechanisms of <i>N. punctiforme</i> in addition to initiation of the scytonemin biosynthesis pathway. </p>

abstract: Desert organisms lead harsh lives owing to the extreme, often unpredictable environmental conditions they endure. Climate change will likely make their existence even harsher. Predicting the ecological consequences of future climate scenarios thus requires understanding how the biota will be affected by climatic shifts. Biological soil crusts (biocrusts) are an important ecosystem component in arid lands, one that covers large portions of the landscape, improving soil stability and fertility. Because cyanobacteria are biocrust’s preeminent primary producers, eking out an existence during short pulses of precipitation, they represent a relevant global change object of study. I assessed how climate scenarios predicted for the Southwestern United States (US) will affect biocrusts using long-term, rainfall-modifying experimental set-ups that imposed either more intense drought, a seasonally delayed monsoon season, or a shift to smaller but more frequent precipitation events. I expected drought to be detrimental, but not a delay in the monsoon season. Surprisingly, both treatments showed similar effects on cyanobacterial community composition and population size after four years. While successionally incipient biocrusts were unaffected, mature biocrusts lost biomass and diversity with treatment, especially among nitrogen-fixing cyanobacteria. In separate experiments, I assessed the effect of rainfall with modified event size and frequency after a decade of treatment. Small, frequent rainfall events surprisingly enhanced the diversity and biomass of bacteria and cyanobacteria, with clear winners and losers: nitrogen-fixing Scytonema sp. benefited, while Microcoleus vaginatus lost its dominance. As an additional finding, I could also show that water addition is not always beneficial to biocrusts, calling into question the notion that these are strictly water-limited systems. Finally, results interpretation was severely hampered by a lack of appropriate systematic treatment for an important group of biocrust cyanobacteria, the “Microcoleus steenstrupii complex”. I characterized the complex using a polyphasic approach, leading to the formal description of a new family (Porphyrosiphonaceae) of desiccation resistant cyanobacteria that includes 11 genera, of which 5 had to be newly described. Under the new framework, the distribution and abundance of biocrust cyanobacteria with respect to environmental conditions can now be understood. This body of work contributes significantly to explain current distributional patterns of biocrust cyanobacteria and to predict their fate in the face of climate change. / Dissertation/Thesis / Doctoral Dissertation Microbiology 2020

Mountains play an important role in providing water resources from snow and ice to downstream urban population centers. In Utah, USA, nearly 86% of the state's population resides in the rapidly growing urban corridor along the Wasatch Front. Water along the Wasatch Front is stored in several deep reservoirs in the Provo River Watershed. Additions of nutrients, into these reservoirs, may stimulate the growth of primary producers (e.g., total phytoplankton and cyanobacteria) potentially attenuating water quality. To determine the influence of nutrients on primary producers in the Provo River's reservoirs, identify whether cyanobacteria are transported downstream, and quantify nutrient thresholds that regulate phytoplankton responses, we monitored and experimentally manipulated total phytoplankton and cyanobacteria in rivers and reservoirs. We found that total phytoplankton biomass (measured as chlorophyll a concentrations) was correlated with the total phosphorus (TP) in the Jordanelle Reservoir and the outflow from the Jordanelle Reservoir. Cyanobacteria biomass (measured as phycocyanin concentration) was correlated with dissolved inorganic N (DIN):SRP in the Deer Creek Reservoir and river sites below the Deer Creek Reservoir. Based on next generation sequencing 16S rDNA of all waters evaluated, the relative abundance of cyanobacteria within bacterial communities was extremely low, with the highest relative abundance of cyanobacteria present in the reservoirs being 10.7% for Deer Creek and 5.5% in Jordanelle during the late summer. Of the 25 genera of cyanobacteria that were found across all waters, five species have the ability to produce cyanotoxin: Microcystaceae; Aphanizomenon MDT14a; Aphanizominon NIES81; and Planktothrix NIVA-CYA. Season determined bacterial community composition in the river and reservoir over the almost two years of sampling with bacterial communities being distinct between the limnetic location in Deer Creek Reservoir to the outflow into the river immediately below the dam. We found no difference between the bacterial communities in the limnetic zone in Jordanelle and the river site directly below Jordanelle Dam. In the nutrient starvation bioassays, cyanobacteria and total phytoplankton responded to the nutrient additions, but there were no specific nutrient thresholds where pigment concentrations leveled out even as nutrient concentration increased. However, when P was added as SRP treatments both total phytoplankton and cyanobacteria concentrations increased, especially in the highest SRP treatment (0.08 mg/L) without any N addition. The addition of N alone did not influence total phytoplankton until there was at least 0.2 mg/L of P added. Further, cyanobacteria required at least 0.2 mg/L of SRP before responding to N levels above 0.8 mg/L. Thus, a nutrient threshold to maintain the reservoir at its current state would be 0.2 mg/L P and between 0.3 and 0.8 mg/L DIN. Our results identify that the water within the reservoirs and rivers are extremely clean and is in no immediate risk of extensive total phytoplankton or cyanobacterial blooms with P being the dominant driver of primary producer activity.

No description available.

Harmful Algae Blooms (HABs) in inland waterbodies (e.g., lakes and ponds) pose serious threat to human health and natural ecosystem. Thus, it is imperative to assess HABs and their potential triggering factors over broader spatiotemporal scales. This study utilizes Chlorophyll-a (Chl-a) concentration in water samples collected from lakes in Illinois as an indirect measurement of HABs. The major objectives were to assess the spatiotemporal pattern of HABs over Illinois regions in recent decades, and to examine different machine learning models for predicting the Chl-a concentration based on publicly available water quality datasets. The Chl-a dataset was compiled from two different sources, the regular monitoring program by Illinois Environmental Protection Agency (IEPA) and the Voluntary Lake Monitoring Program (VLMP), the latter of which was primarily collected by citizen participants. Seven environmental and water quality zones were selected for spatial analyses. Additionally, the temporal patterns were assessed using time-series decomposition of monthly Chl-a concentration datasets. The machine learning pipeline includes two tasks: a regression modeling task for predicting Chl-a concentration, and a classification task for estimating lake trophic status. Different meteorological, land use and land cover, and lake morphometry variables were used as independent variables. Four regression models, i.e., Partial Least Squares Regression (PLSR), Support Vector Machine Regression (SVR), Artificial Neural Network Regression (ANNR), and Random Forest Regression (RFR) were used for the first task of the modeling pipeline, and four classification models, i.e., Logistic Regression Classification (LRC), Support Vector Machine Classification (SVC), Artificial Neural Network Classification (ANNC), and Random Forest Classification (RFC), were used for the second task. Results indicate that: a) the Collinsville region in southwestern part of Illinois exhibited higher mean concentration of Chl-a in its lakes than any other regions from 1998 to 2018; b) the lakes that showed increasing trends in their monthly mean Chl-a concentrations were also clustered in the southwestern region; c) Random Forest outperformed all other models in both classification (Accuracy=60.06%) and regression (R2=38.88%); and d) the land use and land cover variables were found as the most important set of variables in Random Forest models.

Les plastes primaires proviennent d'une cyanobactérie qui a établi une relationendosymbiotique avec un hôte eucaryote. Cet événement a donné naissance au super-groupeArchaeplastida qui inclut les Viridiplantae (algues vertes et plantes terrestres), les Rhodophyta (alguesrouges) et les Glaucophyta. Suite à l'endosymbiose primaire, les algues rouges et vertes ont étendu lacapacité de photosynthèse à d'autres lignées eucaryotes via des endosymbioses secondaires. Bien quedes progrès considérables aient été réalisés dans la compréhension de l'évolution des eucaryotesphotosynthétiques, d'importantes questions sont restées ouvertes, telles que l’identité de la lignéecyanobactérienne la plus proche des plastes primaires ainsi que le nombre et l'identité des partenairesdans les endosymbioses secondaires.Ma thèse a consisté à étudier l'origine et l'évolution précoce des eucaryotes photosynthétiques enutilisant des approches phylogénétiques et phylogénomiques. Je montre par mon travail que les plastesprimaires ont évolué à partir d'un symbiote phylogénétiquement proche de Gloeomargarita lithophora,une cyanobactérie représentant un clade s’étant diversifié précocement et qui a été détectéeuniquement dans les milieux terrestres. Ce résultat fournit des pistes nouvelles sur le contexteécologique dans lequel l'endosymbiose primaire a probablement eu lieu. En ce qui concerne l'évolutiondes lignées eucaryotes avec des plastes secondaires, je montre que les génomes nucléaires deschlorarachniophytes et des euglénophytes, deux lignées photosynthétiques avec des plastes dérivésd'algues vertes, encodent un grand nombre de gènes acquis par transferts depuis des algues rouges.Enfin, je mets en évidence que SELMA, la machinerie de translocation des protéines à travers laseconde membrane externe des plastes rouges secondaires à quatre membranes, a une histoireétonnamment compliquée aux implications évolutives importantes : les cryptophytes ont recruté unensemble de composants de SELMA différent de ceux des haptophytes, straménopiles et alvéolés.Ainsi, ma thèse a permis d’identifier pour la première fois la lignée cyanobactérienne la plus proche desplastes primaires et apporte de nouvelles pistes pour éclaircir les événements complexes qui ontjalonné l’évolution des eucaryotes photosynthétiques secondaires. / Primary plastids derive from a cyanobacterium that entered into an endosymbioticrelationship with a eukaryotic host. This event gave rise to the supergroup Archaeplastida whichcomprises Viridiplantae (green algae and land plants), Rhodophyta (red algae) and Glaucophyta. Afterprimary endosymbiosis, red and green algae spread the ability to photosynthesize to other eukaryoticlineages via secondary endosymbioses. Although considerable progress has been made in theunderstanding of the evolution of photosynthetic eukaryotes, important questions remained debatedsuch as the present-day closest cyanobacterial lineage to primary plastids as well as the number andidentity of partners in secondary endosymbioses.The main objectives of my PhD were to study the origin and evolution of plastid-bearing eukaryotesusing phylogenetic and phylogenomic approaches to shed some light on how primary and secondaryendosymbioses occurred. In this work, I show that primary plastids evolved from a close relative ofGloeomargarita lithophora, a recently sequenced early-branching cyanobacterium that has been onlydetected in terrestrial environments. This result provide interesting hints on the ecological setting whereprimary endosymbiosis likely took place. Regarding the evolution of eukaryotic lineages with secondaryplastids, I show that the nuclear genomes of chlorarachniophytes and euglenids, two photosyntheticlineages with green alga-derived plastids, encode for a large number of genes acquired by transfersfrom red algae. Finally, I highlight that SELMA, the translocation machinery putatively used to importproteins across the second outermost membrane of secondary red plastids with four membranes, has asurprisingly complex history with strong evolutionary implications: cryptophytes have recruited a set ofSELMA components different from those present in haptophytes, stramenopiles and alveolates.In conclusion, during my PhD I identified for the first time the closest living cyanobacterium to primaryplastids and provided new insights on the complex evolution that have undergone secondary plastid-bearing eukaryotes

The MRS/MISS of the Makgabeng Formation encompasses sand cracks, wrinkle marks, mat fragments, mat chips and roll-ups and those of the Magaliesberg formation are wrinkle marks, petees/petee ridges, sand cracks, and multi-directional ripples. The sedimentary process that moderated the formational mechanism of the MISS of the Makgabeng Formation is (descriptively allochthonous) of high energy (inter-dune depositional setting) that eroded, transported and re-deposited mat bound sediments. The genetic mechanism of the MISS of the Magaliesberg Formation is descriptively authochthonous because of enhanced resistance of biostabilized sediments to being reworked. XRF (major and trace) and XRD analysis (qualitative and quantitative) was done on MISS bearing sedimentary rock layers (A) and underlying sedimentary sections (B) of Magaliesberg and Makgabeng samples. Result show high quartz content of all the analyzed samples compared to average sandstones. This premise suggests a relation of microbes (e.g. cyanobacteria) to phototrophy and/photoautotrophy because of the conduction properties of translucent quartz. Also plausible inference is that the intense chemical weathering that produced the quartz arenite was positively influenced by microbes, as noted in some Proterozoic basins. There is higher concentration of Ba in all A samples compared to B (Makgabeng and Magaliesberg) which might be emblematic of biogenicity. The Magaliesberg analyzed samples (MAG 101, 102, 103) exhibit homogeneity by the higher concentration of Al2O3, TiO2, K2O, and P2O5, and lower concentration of SiO2 in the A compared to the B subsamples of a particular sample. Also, Magaliesberg analyzed samples (MAG 101, 102, 103) exhibit homogeneity by the lower concentration of quartz and higher concentration of muscovite in the A compared to the B subsamples. This exact established negative correlation between the duo of SiO2 and quartz, and the quartet of Al2O3, TiO2, K2O, and P2O5, and muscovite as in Magaliesberg samples pertains also to a Makgabeng sample (MKG 102; roll-up). MKG 101 (mat fragment) deviates from this mineralogical and geochemical trend. Each of the A samples of MAG 101, 102, 103, are uniformly of higher concentration in Ce, Cr, Nb, Th, V, Y, Zn, Zr compared to the B version of that sample. MKG 101 and 102 are uniformly of lower concentration of Ce, Cr, Nb, Th, V, Y, Zn, Zr in A compared to the B version of that sample. The A of each of the samples MAG 101, 102, and 103 has higher concentration of Hf and Rb compared to its B; a character that is also exhibit in MKG 102, and MKG 101 is vice versa. Microscopy shows that A of all the samples is of smaller grain size compared to B, espousing affinity of microbes to fine-medium grained sandstones. Microscopy of the Magaliesberg Formation samples show Pseudo petee ridges and pseudo cross lamination which reflect biostabilization, and microscopy of the Makgabeng Formation show roll-ups, mat chips and composite mat chips. The MISS genetic difference of the two formations is related to energy, water residence time (emergence and inundation), Ph, and similarity is related to mutuality in shallow water environment. Mat types are inferred to be biologically, physically and chemically moderated adaptations of microbial communities to specific cum peculiar locally prevailing environmental conditions; factors that are premised on taphonomy and ecology. / Dissertation (MSc)--University of Pretoria, 2014. / tm2015 / Geology / MSc / Unrestricted