Starvation | EPFL Graph Search

Starvation is a severe deficiency in caloric energy intake, below the level needed to maintain an organism's life. It is the most extreme form of malnutrition. In humans, prolonged starvation can cause permanent organ damage and eventually, death. The term inanition refers to the symptoms and effects of starvation. Starvation may also be used as a means of torture or execution. According to the World Health Organization (WHO), hunger is the single gravest threat to the world's public health. The WHO also states that malnutrition is by far the biggest contributor to child mortality, present in half of all cases. Undernutrition is a contributory factor in the death of 3.1 million children under five every year. Figures on actual starvation are difficult to come by, but according to the Food and Agriculture Organization, the less severe condition of undernourishment currently affects about 842 million people, or about one in eight (12.5%) people in the world populati

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

Bacterial cell cycle dynamics: size regulation during exponential growth and role of polyphosphate during starvation response

Aster Louis K Vanhecke

Bacteria are the most diverse and abundant kingdom of life and have adapted to survive and thrive in habitats around the globe. When provided with ample nutrients they grow and divide at staggering rates, increasing their population exponentially. Upon nutrient depletion on the other hand, they quickly adapt by drastically altering their metabolism, halting growth to survive for a very long time. Since bacteria are tiny -about a few micrometers-, visualizing these processes requires microscopy. To measure the dynamics of their shape and inner structures precisely, one needs to choose a technique that balances spatial resolution, temporal resolution and photo-toxicity. In this thesis, I present two projects using advanced dynamic microscopy, first to study cell size regulation during exponential growth in the abundance of nutrients and then to elucidate the role and positioning of polyphosphate granules during cell cycle exit in response to nutrient starvation. During exponential growth, bacteria balance growth and division to regulate their size, resulting in a narrow size distribution, referred to as cell size homeostasis. Recent work tried to uncover what cells sense to decide to divide in order to achieve size homeostasis: time, size, growth or a combination of those. Control of cell division is often equated to control of constriction onset; however, the constriction period still accounts for up to 40% of cell growth and could thus contribute significantly to cell size regulation. We used SIM microscopy to measure constriction kinetics and their impact on cell size regulation in Caulobacter crescentus. We found that constriction rate regulation can determine cell size. Moreover, constriction rate modulation compensates for variability in elongation before constriction, allowing a higher fidelity cell size homeostasis. We suggest a parsimonious model where excess cell wall precursors accumulate proportionally to elongation before constriction and set the rate of constriction. This is the first direct demonstration that constriction rate can contribute to cell size control and homeostasis in bacteria. Upon nutrient starvation, bacteria exit their cell cycle to preserve energy and nutrients. In many bacteria, such as Pseudomonas aeruginosa, this is associated with the accumulation of polyphosphate (polyP) in intracellular granules. PolyP is created by polyphosphate kinases (ppkâs), which are required for successful cell cycle exit and survival of and recovery from long-term starvation. Interestingly, these polyP granules are regularly spaced within the nucleoid. To date, it is not known during which stage polyP is required for cell cycle exit, and what causes the spacing of the granules. Here, we use fluorescence microscopy to probe the cell cycle stage of Îppk cells arrested during nutrient starvation as well as the localization and dynamics of ppkâs. We show that a majority of Îppk cells are arrested with open replication forks. Furthermore, we find that ppkâs localize in distinct patterns, already prior to starvation and polyP granule production, which could be responsible for the positioning of polyP granules. To this end, we developed a background subtraction algorithm to remove cytoplasmic fluorescence, improving accuracy of spot detection and localization.

EPFL2019

WhiB5, a Transcriptional Regulator That Contributes to Mycobacterium tuberculosis Virulence and Reactivation

Hirohito Haruki, Kai Johnsson, Monica Rengifo Gonzalez

The proteins belonging to the WhiB superfamily are small global transcriptional regulators typical of actinomycetes. In this paper, we characterize the role of WhiB5, a Mycobacterium tuberculosis protein belonging to this superfamily. A null mutant was constructed in M. tuberculosis H37Rv and was shown to be attenuated during both progressive and chronic mouse infections. Mice infected with the mutant had smaller bacillary burdens in the lungs but a larger inflammatory response, suggesting a role of WhiB5 in immunomodulation. Most interestingly, the whiB5 mutant was not able to resume growth after reactivation from chronic infection, suggesting that WhiB5 controls the expression of genes involved in this process. The mutant was also more sensitive than the wild-type parental strain to S-nitrosoglutathione (GSNO) and was less metabolically active following prolonged starvation, underscoring the importance of GSNO and starvation in development and maintenance of chronic infection. DNA microarray analysis identified 58 genes whose expression is influenced by WhiB5, including sigM, encoding an alternative sigma factor, and genes encoding the constituents of two type VII secretion systems, namely, ESX-2 and ESX-4.