Bio detection of anoxia in Lake Xochimilco using modified Escherichia - - PowerPoint PPT Presentation

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Jun 04, 2023 107 likes •333 views

Bio detection of anoxia in Lake Xochimilco using modified Escherichia coli by GenetiX Instituto Tecnolgico y de Estudios Superiores de Monterrey, Campus Ciudad de Mxico Our Team Background Xochimilco: Source of water for the

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Bio detection of anoxia in Lake Xochimilco using modified Escherichia coli

by GenetiX

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Instituto Tecnológico y de Estudios Superiores de Monterrey, Campus Ciudad de México

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Our Team

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Background

 Xochimilco:

Source of water for the

Metropolitan Area

Highly touristic and agricultural

element

World Heritage Site
Local endemic species in danger
f extinction
Highly polluted with indutrial

residues

Critical levels of dO2 below

minimun requirements

Artificial eutrophication

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What is Eutrophication?

Dissolved Oxygen Less than 3 mg/L Movement

f water

Temperature Salinity Pressure Photosynthesis Respiration Eutrophication Factors Water Xochimilco 6 mg/L

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Eutrophication Increased Photosynthesis Nutrient Fertilizers Nitrogen and Phosphorus Blocking of light to littoral zone Microbial decomposition Depletion of dissolved oxygen Hypoxia Less than 2.8mg/L Indicators Chlorophyl

Parameter Oligotrophic Eutrophic Average total phosphorus 8.0 84.4 Average total nitrogen 661 1875 Average Chlorophyll a 1.7 14.3 Chlorophyll a. peak concentration 4.2 42.6

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Objective

 Detection of anormal ferric and anoxia/hipoxia

levels on water samples from the Xochimilco Lake by using biosensors

O2Prom 2 ppm>[O2] 2 ppm<[O2] Fe Prom

0.5ppm>[Fe] 0.5ppm<[Fe]

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Topic

 Biosensors:

Fast response in time
Fast and continuous measurement
High specificity (shape-specific

recognition)

Simplicity in its use
Capability of measuring

concentrations ranging from 10-18 to 10-19 M, so we need low sample requirements

Real time measurements

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Justification

Acceptable levels of disolved iron in water: 0.5

ppm

Iron promoter minimal iron dissolution

concentration: 1 ppm

Oxygen Promoter Works in dO2 concentrations
ver 0.2 ppm
dO2 levels in Xochimilco vary around 3 ppm
Minimal requirement of dO2 for wild-life

sustainance 6 ppm

Detection of quantitative values of GFP and RFP

by Flow Cytometry

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Our Biosensor

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Methodology & Results

A B

+ Bacteria Transformation

20 hours 37° C A B A B

A B

Concentrated DNA

A B

Transformed Bacteria Heat Shock

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Methodology & Results

A B

Concentrated DNA

Check +enzymes

+Enzymes Expected Pattern Agarose Gel Cut Bands

A B

Ligation

A B

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Methodology & Results

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Oxygen Promoter + G.F.P Iron Promoter + G.F.P Oxygen Promoter + mR.F.P.

EcoRI Pstl

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Discussions

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Conclusions

Construction of biosensors was not completed due to what appear to be flawed ligations and/or low efficiency competent cells.

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Future Work

 Completion of the biosensors  Proof of concept growing bacteria in absence

and presence of stimuli

 Quantitative measurement of disolved O₂ and

Fe in various concentrations for construction of a photoluminescent scale.

 Application of biosensors to a range of samples

from specific points distributed all along Xochimilco lake.

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Posada Tec 2013

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Travesuras 2014

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Synthetic Biology Conference

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Thanks

 ITESM CCM  Hidrosina  GoNext Project  Tecnologico de Monterrey campus Mty.  Nautilus  Tolerantum  QFB. Isabel Coria

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Special Thanks

 Luis Vaca Dominguez M.D. Ph. D.  Alicia Sampieri García Ph. D.  M.Sc. Lamberto Álvarez-Palacios