Bacterial growth | EPFL Graph Search

Bacterial growth is proliferation of bacterium into two daughter cells, in a process called binary fission. Providing no event occurs, the resulting daughter cells are genetically identical to the original cell. Hence, bacterial growth occurs. Both daughter cells from the division do not necessarily survive. However, if the surviving number exceeds unity on average, the bacterial population undergoes exponential growth. The measurement of an exponential bacterial growth curve in batch culture was traditionally a part of the training of all microbiologists; the basic means requires bacterial enumeration (cell counting) by direct and individual (microscopic, flow cytometry), direct and bulk (biomass), indirect and individual (colony counting), or indirect and bulk (most probable number, turbidity, nutrient uptake) methods. Models reconcile theory with the measurements. Phases In autecological studies, the growth of bacteria (or other microorganisms, as protozoa, microalgae

Co-culture of microalgae and bacteria for wastewater treatment

Thierry Mounir

To address the problem of insucient wastewater treatment and eutrophication of lakes and rivers, the symbiotic association of microalgae and bacteria has shown great potential and numerous advantages. First, co-cultures can exhibit ecient treatment of domestic wastewater, with the combination of high nutrients (nitrogen and phosphorus) removal abilities of the microalgae and ecient degradation of organic matter by bacteria. Second, the symbiosis between microalgae and bacteria exchanging CO2 and O2 makes aeration not necessary, thus signicantly reducing the operating costs of the wastewater treatment plant. Third, recovered biomass of algae could be used as highpro t products like biofuels and food source. Fourth, the xation of the atmospheric CO2 by the microalgae through photosynthesis would contribute to greenhouse gas reduction in the atmosphere. Finally, the decayed microalgal biomass could serve as organic carbon source for denitrication by bacteria. However, this technology is not well-known yet, and further research on the in uence of the operating parameters have to be conducted. Furthermore, its main drawback is the harvesting of the microalgae. Due to their small size, microalgae demonstrate a low settling rate of 3.6 x 10􀀀3 m/h. A solution is to enhance aggregation of the biomass through occulation, followed by granulation to reach fully-developed microalgal-bacterial consortia. These granules exhibit high treatment performances and high settleability. The rst objective of this project was to identify the optimal microalgae:bacteria ratio for COD and nutrients removal. The second objective was to form fully-developed granules, starting from a co-culture, and assess the in uence of dierent agitation speeds on the granulation. The photobioreactor with a microalgae:bacteria ratio of 75:25 expressed the most promising results in COD removal eciency (95% after 8 days), nitrogen removal eciency (94% after 9 days), phosphorus removal eciency (93% after 7 days), and biomass productivity (144 mg/L/d). In this photobioreactor, nitrogen was removed by a combination of assimilation into microalgal biomass and nitrication/denitrication by the bacteria. The COD was removed by degradation by bacteria and mixotrophic microalgae that used the organic matter as carbon source. Finally, phosphorus was removed by assimilation into microalgal biomass. Although obvious aggregation between microalgae and bacteria could be observed, no fully-developed granules were formed after 89 batches of 24 hours. Because of low COD and nitrogen removals, no starvation period was experienced by the co-cultures. Therefore, extracellular polymeric substances were not released in sucient amounts to enhance the granulation. Furthermore, excessive stirring (200 rpm) may have lead to microalgal cells damage, thus promoting bacterial growth over microalgal growth. On the other hand, slower agitation speeds (80 and 120 rpm) seemed to enhance microalgal growth. Overall, the agitation speed appeared to have an indirect impact on the treatment performances and the settleability of the co-cultures, by signicantly in uencing the microalgal and bacterial growths.

2020

The double-edged toxins

Mario Gonzalo Garcia Arraez

Many insect species are associated with endosymbiotic bacteria which have the particularity of living within host tissues. Endosymbionts benefit from this stable and nutritious environment, while providing ecological advantages to their host, such as protection against parasites or thermal tolerance. Spiroplasma poulsonii is an endosymbiotic bacterium that infects natural populations of Drosophila melanogaster. Spiroplasma poulsonii lacks a cell wall, a fact that renders it invisible to the host immune system and allows it to thrive in the host hemolymph. It invades the female germline by co-opting the hostâs yolk transport and uptake machinery, which ensures its vertical transmission. Its efficient transmission is associated with a phenotype called male-killing, whereby infected male embryos die during their early development while infected females survive. The mechanisms that ensure the stability of Drosophila-Spiroplasma symbiosis are increasingly well understood, but the bacterial genes involved remain poorly known because of the intractability of Spiroplasma. This project aims at better characterizing the Drosophila-Spiroplasma interaction, with particular focus on the bacterial side. In the first part, I developed a method to cultivate Spiroplasma poulsonii in vitro by optimizing a commercially available medium. This culture method allowed comparing the transcriptome of in vitro grown versus host-grown Spiroplasma, enabling us to identify putative genes involved in the interaction with the host. Interestingly, inside its insect host, S. poulsonii up-regulates genes coding for toxins of the Ribosomal Inactivating Protein (RIP) family. RIPs were previously known for their role in host protection against macro-parasites, such as wasps and nematodes. Their up-regulation in unparasitized hosts compared to culture was thus peculiar, raising the question what effect these RIPs have on host biology. Thus, in the second part, I studied the function of S. poulsonii RIPs in the absence of parasites. I showed that two of them are constantly expressed within the host and provide evidence that these toxins shorten host life span and increase embryonic mortality. Interestingly, the expression of RIPs was more toxic to male embryos than females, suggesting that RIPs contribute to S. poulsonii-induced male-killing. Last, I studied the D. melanogaster response to RIPs, and how the host mitigates the deleterious effects of these toxins by up-regulating the cytosolic chaperone Heat-Shock-Protein 70B (HSP70B). This protein carries out essential functions in protein homeostasis under normal and stressful conditions such as folding, refolding, or increasing the half-life of proteins. Interestingly, up-regulation of Hsp70B in the presence of RIPs results in an increased lifespan and in a better tolerance to heat stress, which may be ecologically advantageous. Altogether, this work illustrates how Spiroplasma-derived RIP toxins can differentially affect Drosophila depending on the ecological context, ranging from beneficial upon parasite infection or heat shock to detrimental in the absence of such environmental pressures.

EPFL2018

The porous surface model, a novel experimental system for online quantitative observation of microbial processes under unsaturated conditions

Gamze Gülez, Dani Or

Water is arguably the most important constituent of microbial microhabitats due to its control of physical and physiological processes critical to microbial activity. In natural environments, bacteria often live on unsaturated surfaces, in thin (micrometric) liquid films. Nevertheless, no experimental systems are available that allow real-time observation of bacterial processes in liquid films of controlled thickness. We propose a novel, inexpensive, easily operated experimental platform, termed the porous surface model (PSM) that enables quantitative real-time microscopic observations of bacterial growth and activity under controlled unsaturated conditions. Bacteria are inoculated on a porous ceramic plate, wetted by a liquid medium. The thickness of the liquid film at the surface of the plate is set by imposing suction, corresponding to soil matric potential, to the liquid medium. The utility of the PSM was demonstrated using Pseudomonas putida KT2440 tagged with gfp as a model bacterium. Single cells were inoculated at the surface of the PSM, and the rate at which colonies expanded laterally was measured for three matric potentials (-0.5, -1.2, and -3.6 kPa). The matric potential exerted significant influence on colony expansion rates, with a faster rate of spreading at -0.5 than at -1.2 or -3.6 kPa (diameter increase rate, ca. 1,000, 200, and 17 mu m h(-1), respectively). These differences can be attributed to cell motility, strongly limited under the most negative matric potential. The PSM constitutes a tool uniquely adapted to study the influence of liquid film geometry on microbial processes. It should therefore contribute to uncovering mechanisms of microbial adaptation to unsaturated environments.

2008